Conditions: Hypovolemia
Allocation: RANDOMIZED | Intervention model: PARALLEL
Enrollment: 167
Healthy volunteers: no

Brief summary:
This prospective, double-blind randomized controlled trial evaluates the differences in terms of efficacy and safety of gelatin based resuscitation as compared to crystalloid based resuscitation in two parallel groups of patients with severe sepsis / septic shock.

Detailed description:

Arms:
arm 1: Balanced gelatine solution arm 2: Balanced electrolyte solution

Interventions: intervention 1: Balanced gelatine solution intervention 2: Balanced electrolyte solution
Intervention descriptions: intervention 1: Gelaspan combined with Sterofundin ISO intervention 2: Sterofundin ISO

Protocol document:
Prot_002.pdf: 
Page 1 of 10 
 
Study Plan of 
PROSPECTIVE, RANDOMIZED, CONTROLLED, DOUBLE-BLIND, MULTI-
CENTRIC, INTERNATIONAL STUDY ON THE EFFICACY AND SAFETY OF AN 
EARLY TARGET CONTROLLED PLASMA VOLUME REPLACEMENT THERAPY 
WITH A BALANCED GELATIN SOLUTION VS A BALANCED ELECTROLYTE 
SOLUTION IN PATIENTS WITH SEVERE SEPSIS / SEPTIC SHOCK 
 
GENIUS (GELATIN IN ICU AND SEPSIS) 
 
STUDY IDENTIFICATION NO.: HC-G-H-1209 
NCT02715466 
Date of final applicable study plan: 16.06.2020 
 
Marketing Authorization Holder: 
 
B. Braun Melsungen AG 
Germany 
 
 
Organized and Financed by: 
 
B. Braun Melsungen AG 
Division Hospital Care 
Carl-Braun-Str. 1 
34212 Melsungen 
Germany 
 


Page 2 of 10 
 
STUDY OVERVIEW 
 
Title of Study 
PROSPECTIVE, RANDOMIZED, CONTROLLED, DOUBLE-BLIND, 
MULTI-CENTRIC, INTERNATIONAL STUDY ON THE EFFICACY AND 
SAFETY OF AN EARLY TARGET CONTROLLED PLASMA VOLUME 
REPLACEMENT THERAPY WITH A BALANCED GELATIN 
SOLUTION VS A BALANCED ELECTROLYTE SOLUTION IN 
PATIENTS WITH SEVERE SEPSIS / SEPTIC SHOCK 
Acronym 
GENIUS (GELATIN IN ICU AND SEPSIS) 
Investigational Products 
(IPs)  
Investigational test product (gelatin group): 
Gelaspan (plasma adapted gelatin solution for infusion)  
 To be administered in combination with background medication (i.e. 
open-label Sterofundin ISO; a plasma adapted electrolyte solution) in a 
ratio of 1:1 (gelatin:electrolyte) 
 
Investigational reference product (crystalloid group): 
Sterofundin ISO (plasma adapted electrolyte solution)  
 To be administered in combination with background medication (i.e. 
open-label Sterofundin ISO; a plasma adapted electrolyte solution) in a 
ratio of 1:1 
Background Medication 
Open-label Sterofundin ISO (plasma adapted electrolyte solution) 
Phase
IV
Study Design 
Prospective, controlled, randomized, double-blind, international, multi-centric 
study with two parallel groups 
Number of Sites & 
Countries 
Multi-centric, international  
(Participating countries: Austria, Czech Republic, France, Germany, Spain) 
Up to 15 sites
Sample Size 
2 x 304 patients (in total 608 patients) 
Sample size re-calculation after 2 x 200 (in total 400) patients. 
Indication
Hypovolaemia in Severe Sepsis / Septic Shock
Primary Objective 
Investigate the efficacy of early goal directed fluid management of a 
combination of a gelatin and crystalloid regime in comparison to a pure 
crystalloid regime in achieving haemodynamic stability (HDS) in patients 
with severe sepsis / septic shock. 
Primary Variable 
Time until first / initial HDS is achieved (in minutes). 
Time measurement will start with first administration of IPs and will be 
continued until first HDS is achieved.
Secondary Objectives 
Investigation of safety and efficacy parameters of the applied fluid regimes. 
Secondary Variables 
 Safety 
Renal function 
 Serum creatinine (SCr) 
 Serum blood urea nitrogen (BUN) 
 Estimated glomerular filtration rate (eGFR) 
 Need and indication of renal replacement therapy (RRT)  


Page 3 of 10 
 
 Urine output  
 Kidney Disease Improving Global Outcome (KDIGO) score 
 
Coagulation 
 Prothrombin time (PT) 
 Activated partial thromboplastin time (aPTT)  
 International norm ratio (INR) 
Site specific measurements (Innsbruck): 
 Fibrinogen  
 Antithrombin (AT)  
 Platelets absolute 
 
Hepatic function 
 Bilirubin 
 
Adverse Events 
 (Serious) adverse events ((S)AEs) / reactions ((S)ARs) 
 
Other 
 Number of red blood cell (RBC) units 
 Number of fresh frozen plasma (FFP) units 
 Number of other blood products 
 Vasopressor therapy 
 Inotropic therapy 
 
 Efficacy 
Fluid administration according to the volume algorithm 
 Volume needed to achieve first / initial HDS  
 Total volume until 48 h after randomization 
 Administered bottles 
 
Additional administered crystalloid solutions for further volume 
treatment 
 Drug and volume  
 
Fluid balance  
 Fluid intake  
 Fluid output  
 Fluid balance  
 
Haemodynamics 
 Volume responsiveness upon passive leg raising (PLR) or exogenous 
fluid challenge  
 Haemodynamic readings as required by the volume algorithm and 
haemodynamics from 48 h after randomization until ICU discharge or 
day 28, whatever occurs first 
 
Tissue oxygenation and acid base balance 
 Arterial blood gas analyses (BGA) 


Page 4 of 10 
 
 Serum electrolytes 
 Base excess (BE) 
 Lactate / lactate decrease 
 Central venous BGA 
 Arterial oxygen content 
 Oxygen delivery  
 Outcome  
 Length of stay (LOS) in the intensive care unit (ICU) 
 Hospital LOS 
 Days on RRT  
 Number of infection free days 
 Number of antibiotic free days 
 Number of ventilator free days  
 Number of vasopressor free days  
 
 Follow-up 
 Last available SCr (day 28) 
 Colloid therapy (day 28) 
 Mortality & cause of death (if applicable) (day 28 & day 90) 
 Health-related quality of life (HRQoL) (day 90) 
 New RRT / kidney disease (day 90) 
 
 Other variables 
Demographic data  
 Age 
 Gender 
 Height 
 Weight 
 Race 
 Type of patient (e.g. trauma patient, surgical patient) 
 
Anamnesis  
 APACHE II & SOFA  score 
 Temperature 
 Fluid input in the 24 h prior to randomization 
 Origin of sepsis 
 Procalcitonin (optional) 
 Causative organism of infection 
 Medical history 
 
Concomitant medication 
 Antibiotic therapy 
 Nephrotoxic therapy  
 Contrast agents  
 Anticoagulation therapy 
 
Study termination 


Page 5 of 10 
 
Patient Inclusion /  
Exclusion Criteria 
Inclusion criteria: 
 Male or female patients  18 years of age 
 Women of child bearing potential must test negative on standard 
pregnancy test (urine or serum) 
 Patients with body weight  140 kg  
 Patients diagnosed severe sepsis / septic shock (refer to section Fehler! 
Verweisquelle konnte nicht gefunden werden. for definitions) at 
admission on ICU who can be enrolled within 90 min after admission OR 
patients diagnosed severe sepsis / septic shock during ICU stay who can 
be enrolled within 90 min after diagnosis  
 Patients where antibiotic therapy has already been started (prior to 
randomization) 
 Patients who are fluid responsive. Fluid responsiveness is defined as 
increase of > 10% in mean arterial pressure (MAP) after PLR or fluid 
challenge (max. 250 ml crystalloid solution) 
 Signed informed consent by patient, legal representative or authorized 
person or deferred consent 
 
Exclusion criteria: 
 Administration of HES, dextrane solutions or > 500 ml of Gelatin 
solutions within the 24 h prior to randomization 
 Death expected within the next 48 h (moribund patients as defined by 
ASA ≥ class V) 
 Patients with confirmed acute SARS-CoV-2 (COVID-19) infection (as 
available from routine medical records/ patient chart) 
 Patients for whom the need of pressure infusions are expected 
 Requirement for renal support (either continuous or discontinuous 
techniques, including intermittent haemodialysis, haemofiltration and 
haemodiafiltration) 
 Patients receiving therapeutic heparin medication due to chronic 
coagulation disease / anticoagulation medication (i.e. partial 
thromboplastin time > 60 sec) 
 Acutely burned patients (refer to section Fehler! Verweisquelle konnte 
nicht gefunden werden.) 
 Renal failure with oliguria or anuria 
 Severe general oedema 
 Severe congestive cardiac failure 
 Hypersensitivity to the active substance or ingredients of the IPs 
 Hypersensitivity to galactose-α-1,3-galactose (alpha-Gal) or known 
allergy to red meat (mammal meat) and offal 
 Hypervolaemia / hyperhydration 
 Hyperkalaemia  
 Hypercalcaemia  
 Metabolic alkalosis 


Page 6 of 10 
 
 Simultaneous participation in another interventional clinical trial (drugs or 
medical devices studies) 
Investigational Test 
Product 
Gelaspan: Plasma adapted gelatin solution for infusion 
Composition (1000 ml solution contain): 
Succinylated gelatin (= modified fluid gelatin) 
40.0 g 
Sodium chloride 
5.55 g 
Sodium acetate trihydrate 
3.27 g 
Potassium chloride 
0.30 g 
Calcium chloride dihydrate 
0.15 g 
Magnesium chloride hexahydrate 
0.20 g 
 
Excipients for pH-adjustment: 
Sodium hydroxide 40% 
0 – 0.133 g 
Hydrochloric acid 20% 
0 – 0.182 g 
 
Electrolyte concentrations 
Sodium 
151 mmol/l 
Chloride 
103 mmol/l 
Potassium 
4 mmol/l 
Calcium 
1 mmol/l 
Magnesium 
1 mmol/l 
Bicarbonate as Acetate 
24 mmol/l 
Molecular weight, weight average:  
30 000 Dalton 
Molecular weight, number average:  
23 200 Dalton 
Theoretical osmolarity:  
284 mOsm/l 
Investigational Reference 
Product 
 
 
 
Sterofundin ISO: Plasma adapted electrolyte solution for infusion 
Composition (1000 ml solution contain): 
Sodium chloride 
6.80 g 
Potassium chloride 
0.30 g 
Calcium chloride dihydrate 
0.37 g 
Magnesium chloride hexahydrate 
0.20 g 
Sodium acetate trihydrate 
3.27 g 
Malic acid 
0.67 g 
 
Electrolyte concentration 
Sodium  
145.0 mmol/l 
Potassium  
4.0 mmol/l 
Calcium  
2.5 mmol/l 
Magnesium  
1.0 mmol/l 
Chloride  
127.0 mmol/l 
Acetate  
24.0 mmol/l 
Malic acid 
5.0 mmol/l 
pH: 
5.1 – 5.9  
Theoretical osmolarity: 
309 mOsmol/l
Fluid Administration 
Method of Administration 
The administration of IPs and background medication is performed 
intravenously. 
 
 
Dosage  


Page 7 of 10 
 
Dosing of IPs is individualized to the patient’s volume needs. Blinded IP 
volume applied should not exceed 30 ml/kg/day. 
In addition background medication is applied alternating with IPs in a 
ratio of 1:1. 
Fluids are administered according to the volume algorithm as long as the 
patient is fluid responsive. Fluid responsiveness is defined as an increase 
of 10% of MAP and / or stroke volume index (SVI) after endogenous or 
exogenous fluid challenge. An endogenous fluid challenge is achieved by 
means of PLR. An exogenous fluid challenge is achieved by infusing 500 
ml of fluid in 15 min. 
Refer to protocol section Fehler! Verweisquelle konnte nicht gefunden 
werden. for details on the fluid management strategy. 
Duration of Treatment 
Treatment with IPs starts directly with randomization and discontinues 
with the achievement of HDS, achievement of maximum daily dose (30 
ml/kg) or 48 h after randomization, whatever occurs first. 
Visit Schedule 
Prior randomization - 90 min window: 
 Verification of in- / exclusion criteria 
 RBC and antibiotic therapy prior randomization 
 Demographic data including pregnancy test in women of childbearing 
potential 
 Anamnesis & medical history 
At randomization 
 Blood samples for laboratory evaluations (renal function, coagulation, 
hepatic function, lactate, scores) - immediately before first IP 
administration  
Randomisation until 48 h thereafter: 
 Set of catheters (e.g. arterial line, central-venous line) 
 Set of urinary catheter 
 Assessment of HDS and primary endpoint ‘time to first / initial HDS’ 
(lactate, urine production, ScvO2) 
 SCr (every 6 h until 48 h after randomization) 
 Urine output (every 6 h until 48 h after randomization)  
 Blood samples for laboratory evaluations (renal function, coagulation, 
hepatic function) 
 Vasopressor therapy 
 Inotropic therapy 
 Fluid administration (i.e. IP and background medication acc. to volume 
algorithm) 
 Administered crystalloids for further volume treatment 
 Fluid intake & fluid output 
 Fluid responsiveness upon PLR or exogenous fluid challenges as required 
by the volume algorithm 
 Haemodynamic readings as required by the volume algorithm 


Page 8 of 10 
 
 Tissue oxygenation and acid base balance (BGA within the first 6 h every 
2 h, then every 12 h until 48 h after randomization) 
 (Serious) adverse events / reactions 
 Applied blood products  
 Concomitant medications 
 SOFA score, temperature 
 Fulfilment of ICU discharge criteria 
 RRT including indication 
 Invasive mechanical ventilation 
48 h after randomization until ICU discharge or day 28 (whatever occurs 
first): 
 Blood samples for laboratory evaluations (renal function, coagulation, 
hepatic function) 
 Severe sepsis / septic shock (second, third,… hit) 
 Vasopressor therapy 
 Inotropic therapy 
 Administered crystalloids for further volume treatment 
 Fluid intake & fluid output 
 Urine output 
 Haemodynamic readings (minimum and maximum daily)  
 Tissue oxygenation and acid base balance  
 (Serious) adverse events / reactions 
 Applied blood products  
 Concomitant medications 
 SOFA score, temperature 
 Fulfilment of ICU discharge criteria 
 RRT including indication 
 Invasive mechanical ventilation 
 Several outcome parameters (hospital LOS, ICU LOS, number of 
ventilator free days, number of infection free days, number of antibiotic 
free days, number of vasopressor free days, days on RRT)  
 Study termination 
Day 28: 
 Last available SCr documented from ICU discharge until hospital 
discharge or day 28, whatever occurs first  
 Colloid therapy from ICU discharge until hospital discharge or day 28, 
whatever occurs first  
 Mortality & cause of death if applicable 
Day 90: 
 Mortality & cause of death if applicable 
 New RRT / kidney disease 


Page 9 of 10 
 
 HRQoL questionnaire & appendix (if applicable) 
Duration of Study per 
Patient 
Start of study: With randomization 
Treatment period (administration of fluids according to the volume 
algorithm): 
 Start of administration: With randomization  
 Duration of administration:  
 Treatment with fluids according to the volume algorithm 
discontinues with the achievement of HDS or 48 h after 
randomization, whatever occurs first 
 Treatment with additional crystalloid solutions for further volume 
treatment only starts after achievement of HDS, achievement of 
max. daily dose or 48 h after randomization until ICU discharge or 
day 28, whatever occurs first  
End of study: ICU discharge or day 28, whatever occurs first 
Follow ups: day 28 & day 90
Study Schedule 
Planned start: Q1 / 2016 
Planned recruitment time: 5.5 years 
Planned last patient out: Q4 / 2021
Statistical Methods 
Sample size will be based on assumption of difference between the gelatin 
and the crystalloid group. 
Randomization / Blinding  
Randomized with the following strata: 
 Site 
 RBCs pre-treatment within 24 h prior to randomization  
Double-blinded
Data Safety Monitoring 
Board (DSMB)
A DSMB is appointed. 
 
 
 


P
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SAP_001.pdf: 
QMS Document Name: Statistical Analysis Plan 
Page 1 of 33 
Document date: 09JUL2019 
CONFIDENTIAL 
B|BRAUN 
Prospective, randomized, controlled, double-blind, multi-centric, 
international study on the efficacy and safety of an early target-controlled 
plasma volume replacement therapy with a balanced gelatin solution vs. a 
balanced electrolyte solution in patients with severe sepsis / septic shock 
GENIUS (GELATIN IN ICU AND SEPSIS) 
Statistical Analysis Plan 
Final Analysis 
Version: Final 2.0 
Date: 05JUL2022 
Study Identification No: HC-G-H-1209 
     
NCT02715466 
 
EudraCT No: 2015-000057-20  




 
 
Version: 2.0 
Version Date: 05JUL2022 
Statistical Analysis Plan: final analysis  
 
 
QMS Document Name: Statistical Analysis Plan 
Page 3 of 33 
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CONFIDENTIAL 
 
Table of Contents 
 
 
1 
Introduction .................................................................................................................................... 5 
1.1 
Preface ................................................................................................................................... 5 
1.2 
Timing of statistical analyses ................................................................................................. 5 
2 
Modification History ....................................................................................................................... 5 
2.1 
Changes to the study protocol .............................................................................................. 5 
2.2 
Changes to previous SAP versions ......................................................................................... 6 
3 
Study Design.................................................................................................................................... 6 
3.1 
Sample size estimation .......................................................................................................... 7 
3.2 
Randomisation, blinding and unblinding procedures ........................................................... 8 
4 
Analysis Sets .................................................................................................................................... 8 
5 
General Statistical Methods and Definitions .................................................................................. 9 
5.1 
General statistical methods .................................................................................................. 9 
5.2 
Assignment to analysis/treatment groups .......................................................................... 10 
5.3 
Covariates and strata .......................................................................................................... 11 
5.4 
Subgroups ............................................................................................................................ 11 
5.5 
Missing data ........................................................................................................................ 12 
5.6 
Observation and analysis times........................................................................................... 13 
5.7 
Multiple Comparison/Multiplicity ....................................................................................... 13 
6 
Patient Accounting and Disposition .............................................................................................. 13 
6.1 
Patient accounting .............................................................................................................. 13 
6.1 
Disposition and withdrawals ............................................................................................... 13 
6.2 
Major protocol deviations ................................................................................................... 14 
7 
Demographics and Background Characteristics ........................................................................... 14 
7.1 
Demographics...................................................................................................................... 14 
7.2 
Disease characteristics ........................................................................................................ 15 
7.3 
Medical history .................................................................................................................... 15 
8 
Exposure ........................................................................................................................................ 16 
8.1 
Treatment groups ................................................................................................................ 16 
8.2 
Dosage and Treatment duration ......................................................................................... 16 
9 
Concomitant Therapies ................................................................................................................. 17 
9.1 
Not allowed concomitant therapies .................................................................................... 17 


 
 
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10 Efficacy .......................................................................................................................................... 18 
10.1 Primary efficacy analysis ..................................................................................................... 18 
10.2 Secondary efficacy analyses ................................................................................................ 19 
10.3 Other secondary analyses ................................................................................................... 20 
10.4 Exploratory analyses ........................................................................................................... 21 
11 Safety ............................................................................................................................................ 22 
11.1 Adverse events .................................................................................................................... 22 
11.2 Vital signs ............................................................................................................................. 23 
11.3 Clinical safety laboratory ..................................................................................................... 23 
11.4 Kidney Disease Improving Global Outcome (KDIGO) score ................................................ 24 
12 Sensitivity Analyses ....................................................................................................................... 24 
13 Follow-up analyses ........................................................................................................................ 25 
13.1 Follow-up therapies ............................................................................................................ 25 
13.2 Health Related Quality of Life (HRQoL) 90 days after randomization ................................ 25 
14 Interim Analysis ............................................................................................................................. 25 
15 Statistical Analyses for Safety Monitoring .................................................................................... 25 
16 Software ........................................................................................................................................ 26 
17 Abbreviations ................................................................................................................................ 27 
18 Appendices .................................................................................................................................... 29 
18.1 APACHE II score ................................................................................................................... 29 
18.2 SOFA score ........................................................................................................................... 29 
18.3 SAPS II score ........................................................................................................................ 30 
18.4 KDIGO score ........................................................................................................................ 31 
19 References .................................................................................................................................... 33 
 
 
 


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1 
Introduction 
1.1 
Preface 
The objective of this document is to detail the statistical methodology to be used for the final statistical 
analysis of the GENIUS study (Gelatin in ICU and Sepsis).  
The statistical analysis plan is based on the following information and documents: 
Study Protocol  
2015-06-26 – Version 1.0 
Amendments to Study Protocol 2015-11-30 – Version 2.0 incorporating local Amendment No. 1 - 
Austria dated 2015-09-30, local Amendment No. 1 - Czech 
Republic dated 2015-09-22, local Amendment No. 1 - Germany 
dated 2015-10-09, global Amendment No.1 dated 2015-11-30 
2017-05-02 – Version 3.0 incorporating global Amendment No.2 
dated 2017-05-02 
2018-07-20 – Version 4.0 incorporating global Amendment No.3 
dated 2018-07-20 
2020-01-10 – Version 5.0 incorporating global Amendment No. 
4.0 dated 2020-01-10 
2020-07-16 – Version 6.0 incorporating global Amendment No. 
5.0 dated 2020-07-16 
1.2 
Timing of statistical analyses 
The following statistical analyses are planned for this study: 
▪ 
Final analysis: the final unblinded analysis will take place after last patient out, the Blind Data 
Review Meeting (BDRM) is performed, and the database is locked. 
2 
Modification History 
2.1 
Changes to the study protocol 
The first version of the statistical analysis plan (SAP) corresponded to protocol version 6.0.  
Due to the premature termination of the study recommended by the data safety monitoring board 
(DSMB) on 8th of December 2021, the interim analysis (IA) for sample size recalculation (SSR) as 
planned in the study protocol is obsolete and thus only the final analysis of the study will be presented. 
The study populations defined on protocol version 6.0 section 14.8 are adapted in this document. The 
full analysis set was replaced by the safety set and the per protocol set is not taking into account the 
criteria of stopping of treatment due to adverse reaction. 
Moreover, the time unit used for endpoints as the time to HDS will be changed from minutes to hours 
because it is expected that no patient is stabilized within minutes. 


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More subgroup analyses were included in section 5.4 of this SAP. 
Additional changes to the analyses planned in the study protocol were included in this second version 
of the SAP, as described in section below. Main updates were related to the discussions during the 
Blind Data Review Meeting (BDRM). 
2.2 
Changes to previous SAP versions 
This is the second version of the SAP. The following main changes were implemented compared to 
first version dated 12Jan2022: 
• 
More details were added about the major protocol deviations (PDs) defined during the BDRM 
and the analyses to be repeated on the Per Protocol (PP) set. Details are specified in section 4 
and section 6.2. One subgroup was added as a result of the discussions in the BDRM, please 
refer to section 5.4. Additionally, categories for SOFA were updated after the BDRM. 
• 
Some treatment comparisons for secondary efficacy analyses were removed due to the high 
quantity of missing data, please refer to section 10.2. Additionally, MANOVA analyses 
specified in section 12 were removed due to the same reason. 
• 
The calculation of the total score for the Acute Physiology and Chronic Health Evaluation II 
(APACHE II), the Sequential Organ Failure Assessment (SOFA), and the Simplified Acute 
Physiology Score II (SAPS II) was modified including missing values rules. Updates are specified 
in the respective appendices. 
• 
Summary statistics on overall dosing for concomitant medications was removed, expect for 
crystalloid solutions, due to a lack of standardization to collect the doses in the electronic case 
report form (eCRF). 
• 
Kidney Disease Improving Global Outcome (KDIGO) score calculation was adapted considering 
only serum creatinine values without urine output. 
• 
Additionally minor modifications on outputs layout were included to improve the 
interpretation of the study results.  
3 
Study Design 
Indication 
Hypovolaemia in Severe Sepsis / Septic Shock 
Design 
Prospective, 
randomized, 
controlled, 
double-blind, 
multi-centric 
international study with two parallel groups 
Phase 
IV 
Primary Objective 
To investigate the efficacy of early goal directed fluid management of a 
combination of a gelatin and crystalloid regime in comparison to a pure 
crystalloid regime in achieving haemodynamic stability (HDS) in patients 
with severe sepsis / septic shock 


 
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Secondary Objective 
Investigation of safety and further efficacy parameters of the applied fluid 
regimes 
Treatments 
Investigational medicinal test product (gelatin group): 
Gelaspan (plasma adapted gelatin solution for infusion) 
To be administered in combination with background medication (non 
investigational medicinal product: open-label Sterofundin ISO; a plasma 
adapted electrolyte solution) in a ratio of 1:1 (gelatin:electrolyte) 
 
Investigational medicinal reference product (crystalloid group): 
Sterofundin ISO (plasma adapted electrolyte solution) 
To be administered in combination with background medication (non 
investigational medicinal product: open-label Sterofundin ISO; a plasma 
adapted electrolyte solution) in a ratio of 1:1 
Number of patients 
N = 608 patients with n=304 patients in the Gelatin and Crystalloid group, 
respectively. 
Planned enrolment 
Participating countries: Austria, Czech Republic, France, Germany, Spain, 
Hungary. 
Up to 15 sites 
3.1 
Sample size estimation 
The primary variable is the time to first / initial HDS, and it was used to calculate the sample size. The 
data of a study investigating a hydroxyethyl-starch (HES) solution (Guidet et al., 2012; CRYSTMAS-trial) 
served as the basis for the sample size calculation. It was however a comparison of a HES-solution with 
sodium chloride and the results were only an orientation and therefore an interim analysis for sample 
size recalculation was planned which is obsolete due to the premature termination of the study by the 
DSMB.  
Sample size calculation using the software nQuery and nTerim 2.0 resulted in a sample size of 608 
patients in total. It was based on the following assumptions:  
• 
effect size (difference of means / common standard deviation) = 0.25 with difference in means 
= - 2.5 h and common standard deviation = 10 h  
• 
error of 1st kind α = 5% (two-sided)  
• 
power = 80%  
Applying a t- test for group comparison yielded a sample size of 253 patients per group for HDS in 
minutes. Taking a drop out rate of 20% into account, a sample size of 304 patients per group (i.e. 608 


 
 
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patients in total) is needed. Drop-out patients are defined as those patients who do not receive any 
investigational product.  
Withdrawn patients who did not receive any Investigational product (IP) will not be replaced because 
they are considered in the dropout rate. 
3.2 
Randomisation, blinding and unblinding procedures 
Patients were randomized to either treatment in a 1:1 ratio. 
The list of treatment assignments considering the stratification for site and red blood cell (RBC)s pre-
treatment were generated by a statistician (not involved in the study data analyses) comprised of 
consecutive blocks with the order of assignments chosen at random (e.g. random permuted block of 
size 4, 6 or 8). The randomization lists (initial and new sites) were prepared prior to the initiation of 
the study site and remained with the statistician. A signed copy of the randomization lists was  given 
to the qualified person for IPs (sponsor), who was responsible for the blinding of the samples. Based 
on the randomization lists the statistician also issued sets of emergency envelopes for each 
participating site for emergency unblinding. The sponsor, i.e. qualified person study supply and drug 
safety officer received a complete set of emergency envelopes for emergency situations. In addition, 
the biometrician of the Data Safety Monitoring Board (DSMB) received the de-blinding envelopes for 
each patient, in case un-blinding upon request of the DSMB was necessary. The same procedure would 
be performed in case of new study sites and recruitment of more patients per sites as initially 
expected.  
After closure of the database and determination of the analysis populations (in a blinded data review 
meeting if appropriate) the study will be unblinded.  
The population was stratified with regard to site and RBC pre-treatment 24 h prior to randomization. 
These stratification variables will be included as covariate in the primary analysis (according to 
CPMP/EWP/2863/99). 
4 
Analysis Sets   
The statistical analyses and presentation of data will be based on the following analysis sets: 
• 
Enrolled Analysis Set 
It includes all patients with an informed consent signed by patient, legal representative or 
authorized person or deferred consent according to local legislation / requirements approved 
by the ethic committee / competent authority. 
• 
Intent-To-Treat Set (ITT)  
The ITT set includes all patients randomized. Following the intent-to-treat (ITT) principle all 
patients will be analyzed according to the treatment group to which they were randomized. 
This population will be used for the description of demographics and other baseline 
characteristics, and for the assessment of efficacy endpoints. 
• 
Safety Analysis Set (SAF)  


 
 
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The SAF includes all ITT patients treated at least once with the study medication. All patients 
will be analyzed according to the treatment they received. This population will be used mainly 
for the safety endpoints and exposure analyses. 
• 
Per Protocol Set (PP) 
The PP set includes all patients from the SAF set, excluding patients with major protocol 
deviations (please refer to section 6.2). All patients will be analyzed according to the 
treatment they received. The assessment of protocol deviations and the categorization into 
major and minor deviations was performed before unblinding during the BDRM. As the 
difference between the PP set and the SAF set is higher than 10% of the patients included in 
SAF, main analyses based on SAF will be repeated in the PP set. As the ITT and the SAF set 
contain the same number of patients for this study (all patients randomized were treated), 
the difference between the PP set and the ITT set is the same, also greater than 10% of the 
patients included in ITT. Therefore, main analyses based on ITT will be repeated in PP set. 
Minor protocol deviations do not lead to the exclusion of patients from any analysis set.  
If not otherwise stated in the respective section, the statistical analyses will be performed on the 
following analysis sets: 
Analyses 
Enrolled 
ITT 
SAF 
PP 
Disposition 
✓ 
 
 
 
Demographics and background characteristics 
 
✓ 
 
(✓) 
Exposure and compliance 
 
 
✓ 
 
Concomitant therapies 
 
 
✓ 
(✓) 
Primary Efficacy analysis  
 
✓ 
 
(✓) 
Secondary Efficacy analyses 
 
✓ 
 
(✓) 
Other/Exploratory Efficacy analyses 
 
✓ 
 
(✓) 
Safety analyses 
 
 
✓ 
(✓) 
The symbol (✓) means that the related analyses were optional at the planning of the analyses.  
The following analyses will be repeated for the PP set as decided during BDRM discussion: 
• 
Demographics and background characteristics 
• 
Concomitant therapies 
o Antibiotics 
o Colloid therapy (not allowed medication) 
• 
Primary efficacy analyses 
• 
Other / exploratory efficacy analyses 
• 
Safety analyses 
Specific TLGs to be repeated on the PP set are included in Table Shells. 
5 
General Statistical Methods and Definitions 
5.1 
General statistical methods 


 
 
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The statistical analyses will be presented by treatment group for the different analysis sets as defined 
in section 4. 
In general, continuous variables will be summarised using descriptive statistics, i.e. generally 
displaying number of patients in the respective analysis population, number of patients with data, 
mean, standard deviation, minimum, lower quartile, median, upper quartile, and maximum.  
Categorical variables will be summarised by using frequency counts and percentages.  
In case at least one parameter in a table has missing values, a row/column with the number of missing 
values will be added. In case no missing values are documented at all for a table, no missing 
rows/columns will be presented, even if missing rows/columns are foreseen in the respective table 
shell.  
Individual patient data listings will be provided for all data collected. Listings will be sorted by patient 
ID, and when appropriate by visit or other identifiers for sequence or type of observation. All listings 
will present the patient ID, site, treatment group, strata, age, and gender. 
Means and medians will be presented with one additional decimal place and standard deviation will 
be presented with two additional decimal places compared to the original source data. Minimum and 
maximum values will be presented using the same number of decimal places as the patient data. 
Percentages will be presented with one decimal place if not otherwise stated.  
If the number of patients in a category is 0, then percentage will not be displayed, and only a count of 
0 will be shown.  
P-values will be reported to 4 decimal places at least. Values less than 0.0001 will be displayed as 
<0.0001. Values above 0.9995 will be displayed as 1.0000. 
If not otherwise specified, all statistical tests will be two-sided with a significance level of 5%.  The 
confidence level for the calculation of confidence intervals will be 95%, respectively.  
5.2 
Assignment to analysis/treatment groups 
Treatments will be labelled as follows: 
• 
Gelatin group 
• 
Crystalloid group 
• 
Total (when applicable) 
They are defined as: 
• 
Gelatin group: 
 
 
• 
Crystalloid group: 


 
 
   
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5.3 
Covariates and strata 
The population will be stratified with regards to site and RBC pre-treatment 24 h prior to 
randomization (yes/no). These stratification variables will be included as covariate in the primary 
analysis, as will be specified in section 10.1. Frequencies of randomized patients within each stratum 
will be presented as part of the analysis. 
5.4 
Subgroups 
The following subgroup analyses are planned to be conducted for the primary endpoint: 
• 
Strata (prior RBC treatment, sites) 
• 
Prior RBC treatment as confirmed in the eCRF and not according to assigned stratum 
• 
Administration of fluids 24 h prior randomisation (yes/no) 
• 
Acute Physiology And Chronic Health Evaluation II (APACHE II) total score (algorithm specified 
in appendix 18.1): < 20 score points, 21-30 score points, > 30 score points. Final categories 
were confirmed at the BDRM. 
• 
Sepsis-related Organ Failure Assessment (SOFA) total score at baseline (algorithm specified in 
appendix 18.2): < 10 score points, ≥ 10 score points. Final categories were re-defined at the 
BDRM. 
• 
Renal assessment from SOFA at baseline: 1-2 vs 3-5. Score 1 is defined as the recorded score 
0 in the eCRF that corresponds to the category “Creatinine < 110 umol/L (1.2mg/dL) or urine 
output >=500 mL/d”. Score 2 corresponds to the recorded score 1 in the eCRF, and so forth. 
• 
Administration of blood products during the study (yes/no) 
• 
Septic shock / severe sepsis at diagnosis  
• 
One occurrence of septic shock or severe sepsis / at least two occurrences of septic shock or 
severe sepsis (episodes of severe sepsis / septic shock) 
• 
Diagnosis of severe sepsis or septic shock at intensive care unit (ICU) admission / diagnosis of 
severe sepsis or septic shock during ICU stay 
• 
Country 
• 
Concomitant medication (allowed / not allowed). Please refer to section 9 for further details 
on the definition of allowed and not allowed concomitant medication. Under allowed 
medication, only patients that didn’t receive any not allowed medications (i.e. that received 
only allowed medications) will be included. Not allowed medication subgroup will include 
patients that received at least one dose of not allowed medication as described in section 9. 
• 
Use of renal replacement therapy (RRT) during the study (yes, no) 


 
 
   
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• 
Type of patient (trauma, surgical, medical) 
In case a subgroup category has less than 10 patients, descriptive data will be presented but treatment 
comparisons will not be made for that category. 
5.5 
Missing data 
Every effort will be made to collect all data points in the study. Missing data will not be imputed. 
Partial and missing dates will be imputed as specified below only to determine if an event is 
concomitant to the study drug administration or not, assuming the most conservative approach. 
Missing data are only those which are required as defined in the protocol and had not been recorded 
in the eCRF during study participation of a patient. For example, in case a patient terminated the study 
on day 7, all data thereafter are not counted as missing.  
Partial or missing dates and times 
Where the start date of an adverse event or medical condition (administration of a concomitant 
medication needed) is missing or partially missing, the event will be assumed to be started after the 
start of study medication, unless there is clear evidence (through comparison of partial dates/times) 
to suggest otherwise. Duration of an event with missing start or end date will remain as missing. 
In case of a partial or missing end date the medical condition will be assumed to be ongoing after start 
of study medication except if the partial date indicates that the condition stopped prior to start of 
study medication.  
Remaining missing dates will be imputed as follows: 
For event start date, imputation rule is to conservatively capture events with missing start dates as 
occurred during treatment: 
• 
If “day” is the only missing field, impute the “day” as the first dose date if their “month” are 
the same. Otherwise impute the “day” as the first day of the month documented. 
• 
If “day” and “month” are the only missing fields, impute the “day” and “month” as the first 
dose date if their “year” is the same. Otherwise impute the “day” and “month” as the first of 
January of the year documented. 
• 
If “day”, “month”, and “year” are all missing, to be conservative, the event will be assumed to 
occur on the same day as the first dose was administered. 
For event end date, imputation rule is to conservatively capture events with missing end dates as 
occurred during treatment: 
• 
If “day” is the only missing field, impute the “day” as the last day of the month documented if 
their “month” are the same. 
• 
If “day” and “month” are the only missing fields, impute the “day” and “month” as December 
31 of the year, if the patient is alive at that date. Otherwise impute the date of death. 
• 
If “day”, “month”, and “year” are all missing, to be conservative, the event will be assumed to 
be ongoing at the end of the study for the respective patient.  
Dates as collected will be presented in data listings. 
Outliers 
No outliers were identified for the primary endpoint variable ‘time to HDS’. 


 
 
   
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For laboratory parameters, as specified in section 11.3, box plots by visit will be performed for the 
change from baseline in order to detect the outliers. Outliers were detected and checked by data 
management, and specific box plots for serum creatinine will be repeated excluding change from 
baseline values greater or lower than 200 umol/L (as agreed during the BDRM). 
5.6 
Observation and analysis times 
Study days  
Study day is defined as the number of days since first randomized treatment administration, and is 
calculated as: 
Study day = assessment date – date of first randomized treatment administration + 1. 
Therefore, the date of the first administration of randomized treatment will be Day 1. 
Study time  
Study time is defined as the number of minutes since the time of the first randomized treatment to 
the assessment time. The study time in hours will also be derived. 
Definition of baseline values or first assessment 
The baseline value is defined to be the last value which was assessed before first study drug 
administration.  For the parameters where values were not measured / available at baseline, the value  
of the first assessment measured/available shortly after treatment start will be used. 
5.7 
Multiple Comparison/Multiplicity 
All tests will be conducted with an error α= 0.05 two-sided, unless otherwise specified. 
Tests of all secondary variables will be carried out in the area of exploratory data analysis if applicable. 
Therefore, corresponding p-values are to be regarded as exploratory ones and no adjustments for 
multiple testing will be made. 
6 
Patient Accounting and Disposition 
6.1 
Patient accounting 
The number and relative frequencies of patient in each analysis set as defined in section 4 will be 
presented overall and by randomized treatment (if applicable), and will be listed. Additionally, the 
number of patients randomized per country and site will be presented by treatment group. 
Eligibility criteria will be presented in a data listing, containing the information of the informed 
consent(s). 
6.1 
Disposition and withdrawals 
The number and relative frequency will be presented by treatment group based on all enrolled 
patients for 
▪ 
screening phase patients, eligible patients, and failures 
▪ 
patients randomized 


 
 
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▪ 
first treatment phase and second treatment phase patients, each separately, and both phases 
▪ 
patients who discontinued the treatment due to AE 
▪ 
patients who prematurely discontinued the study, related reasons, and person who took the 
decision 
▪ 
patients who achieved HDS and not, and patients with missing HDS status 
▪ 
patients who entered and completed follow-up 1, follow-up 2, and follow-up 1 and 2 
▪ 
patients that completed the study 
▪ 
patients with treatment code broken  
A listing with disposition information will be presented for all enrolled patients. Additionally, a 
disposition table by country will be presented. 
6.2 
Major protocol deviations 
A protocol deviation (PD) is any noncompliance with the study protocol, Good Clinical Practice (GCP), 
or Manual of Procedures (MOP) requirements. The PD which impacts the interpretability of the main 
study results and assessment of the treatment effect will be marked as major protocol deviation. A 
pre-selection of all protocol deviations was reviewed and finally classified as either major or minor 
during the BDRM. Please refer to BDRM minutes for a final assessment on those. 
Major PDs were identified for the following categories:  
• 
Concomitant medication 
• 
In/Exclusion criteria 
• 
IP storage 
• 
IP treatment/Volume algorithm 
• 
Primary endpoint 
• 
Randomization 
The number and relative frequency of patients with major protocol deviations leading to exclusion 
from the Per Protocol set will be presented by treatment group for all patients of the ITT. Additionally, 
patients with major protocol deviations related to Covid-19 pandemic situation will be also presented. 
A listing with all protocol deviations will be presented based on ITT set. 
7 
Demographics and Background Characteristics 
Demographic and baseline characteristics as specified in detail below will be presented descriptively 
by treatment group based on ITT set.  
7.1 
Demographics 
The following demographic characteristics will be presented by frequencies or summary statistics, as 
applicable: 
• 
Gender (male/female) 
• 
Age (years) as entered in the electronic case report form (eCRF) 
• 
Age ranges (18-64, 65-84, ≥85 years) 


 
 
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• 
Race (Black or African American, American Indian or Alaska Native, Asian, Native Hawaiian or 
Other Pacific Islander, White) 
• 
Weight (kg) at baseline 
• 
Height (cm) at baseline 
• 
Body mass index (BMI) (kg/m2) at baseline as automatically calculated within the eCRF  
• 
Body surface area (BSA) at baseline [m2]: 0.007184 𝑥 𝑊𝑒𝑖𝑔ℎ𝑡0.425 [𝑘𝑔] 𝑥 𝐻𝑒𝑖𝑔ℎ𝑡0.725[𝑐𝑚] 
• 
Temperature (C) overall, minimum and maximum at baseline 
• 
Childbearing potential data 
A data listing containing the mentioned demographic variables will be presented based on ITT set. 
7.2 
Disease characteristics 
A frequency table displaying the type of patient (trauma, medical or surgical patient) will be provided 
together with information on the type of sepsis diagnosed. The time (min) from the time of admission 
to ICU to randomization and the time (min) from diagnosis to ICU admission will be also derived and 
presented descriptively for those patients where applicable. The time from diagnosis to randomization 
will also be summarized for patients that are on the ICU at the time of trial participation, i.e. diagnosis 
of severe sepsis/septic shock on ICU and following randomization. 
Additionally, summary statistics and frequencies will be provided for the following parameters of 
anamnesis: 
• 
Simplified Acute Physiology Score II (SAPS II) score, APACHE II, and SOFA total score at 
baseline. Please refer to appendices 18.1, 18.2, and 18.3 for the specification of the APACHE 
II, SOFA, and SAPS II score calculation respectively. 
• 
Fluid input in the 24 h prior to randomization: yes/no and amount (ml)  
• 
Antibiotic therapy prior to randomization: yes/no  
• 
RBC therapy 24 hours prior randomization: yes/no 
• 
Lactate prior treatment (mmol/L) 
• 
Fluid responsiveness for inclusion: type of test (Fluid challenge (FC) or Passive leg raising 
(PLR)), volume (ml) by type of test, Mean arterial pressure (MAP) change (%), MAP before 
PLR/FC (mmHg), MAP after PLR/FC (mmHg) 
A correlation analyses among SAPS II, APACHE II and SOFA total scores at baseline will be done thought 
a correlation test. Pearson correlation coefficients and related p-values will be tabulated. 
Additional details on disease characteristics parameters, as origin of sepsis or causative organism of 
infection, will be provided in a data listing. Details of fluid input in the 24h prior to randomization and 
antibiotic therapy prior to randomization will also be presented in data listings. 
7.3 
Medical history 
Past and ongoing medical history is be coded using the Medical Dictionary for Regulatory Activities 
(MedDRA®), version 24.1, and will be summarized presenting numbers and frequencies by primary 


 
 
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System Organ Class (SOC) and Preferred Term (PT) by treatment group. Past medical history are the 
disorders not ongoing during the study conduct.  
If patients have more than one disease within a SOC or PT they will be counted only once for the 
respective SOC or PT. 
8 
Exposure  
The SAF set will be used to present exposure analyses. 
8.1 
Treatment groups 
Treatments will be labelled as follows: 
• 
Gelatin group 
• 
Crystalloid group 
• 
Total 
The number and percentage of patients will be presented by treatment group based on SAF set. 
Treatment-related summaries will be presented separately for the IP (first and second treatment 
phase together) and the background medication (BG).  
8.2 
Dosage and Treatment duration 
The IP treatment is defined for two treatment phases: 
The first treatment phase starts with randomization and ends in general at the time a haemodynamic 
monitoring system is in place or HDS has already been achieved and confirmed. 
If the HDS criteria are not fulfilled during this treatment phase until having a haemodynamic 
monitoring system in place the patient enters the second treatment phase. For this a haemodynamic 
monitoring system able to directly measure cardiac output (i.e. Stroke volume index (SVI)) has to be 
in place. First treatment phase is not needed when a hemodynamic system is already in place at IP 
treatment start. 
The second treatment phase starts with the time a haemodynamic monitoring system is in place and 
ends when HDS occurs for the first time or 48 hours after randomization, whatever occurs first. 
Treatment with blinded IPs in combination with unblinded background medication ends with the 
achievement of first / initial HDS, achievement of maximum daily dose of 30 ml/kg for the IPs or after 
48 h after randomization, whatever occurs first.  
The total IP/BG treatment duration will be calculated in hours as the complete time of the two 
treatment phases:  
Total Treatment duration (hours) = min(time(confirmed* HDS, maximum daily dose, 48 h after 
randomization))-time(treatment start)  
*confirmed covers the time from initial HDS until end of the 4 hours confirmation period. 
Summary tables by treatment group will be provided as applicable for the following variables:  
• 
Total treatment duration (hours) by IP and BG and total 


 
 
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• 
Total volume of study fluids administered according to the volume algorithm needed to 
achieve first HDS [ml] by IP and BG and total 
• 
Total volume of fluids over study time [ml] (max. until 48 h after randomization) by IP and BG 
and total 
• 
Mean volume of fluid bottles per fluid administration [ml] (IP, BG, total) 
• 
Total of administered bottles by IP and BG and total 
• 
Total volume of additional administered crystalloid solutions [ml] 
• 
Fluid intake [ml] 0-24h and 24-48h 
• 
Fluid output [ml] 0-24h and 24-48h 
• 
Fluid balance = Fluid input [ml] – fluid output [ml] 0-24h and 24-48h 
• 
Fluid overload [%] = [(fluid intake – fluid output)/weight] x 100 0-24h and 24-48h 
• 
Total number of drug interruptions/stops/changes due to AE 
A data listing including the fluid administrations according to volume algorithm will be presented for 
the first and second treatment phase. Similarly, a data listing including information about the 
assessment of fluid responsiveness upon PLR/MAP and haemodynamic monitoring/SVI will be 
presented. 
9 
Concomitant Therapies 
Concomitant medications are coded according to the World Health Organization drug dictionary 
(WHO-Drug version 2021Q2_B3_Global) and stored with generic (preferred) names.  
Medications that were taken after the first dose of randomized treatment will be considered 
concomitant medications. Missing or partly missing stop dates will be imputed using the rules defined 
in section 5.5. The number and frequency concomitant medications will be given per medication 
preferred name based on SAF set. If a patient has received more than 1 drug within a preferred name, 
he/she will be counted only once for this preferred name. 
Additionally, overall duration for each concomitant therapy will be summarized, and total volume for 
crystalloid solutions therapy will be also summarized. 
One concomitant medication table per type of therapy will be presented: antibiotic agents, 
anticoagulation agents, blood products, contrast agents, crystalloid solutions, inotropic agents, 
nephrotoxic agents, vasopressors, other concomitant medication. 
Data listings for concomitant medications will be also presented indicating administration and dosing 
details. 
9.1 
Not allowed concomitant therapies 
The following concomitant medications are not allowed for this study (please refer to section 9.2 of 
the study protocol): 
• 
During the treatment period, that is between randomization and until the achievement of HDS 
or 48 h after randomization, whatever occurs first, administration of other colloid volume 
replacement agents (e.g. other gelatin solutions, albumin, dextran, HES solutions) aside from 
colloids to be administered according to the volume algorithm is NOT allowed.  


 
   
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• 
Beyond the treatment period, that is between HDS or 48 h after randomization and ICU 
discharge or day 28, whatever occurs first administration of colloid volume replacements (i.e. 
gelatin solutions, albumin, dextran, HES solutions or other colloid solutions) is NOT allowed; 
i.e. only crystalloid solutions are allowed in this study period.  
Final classification of allowed and not allowed medications was defined during the BDRM. Not allowed 
medications leading to the corresponding exclusion from the PP set are:  
• 
Colloid solutions (including Gelatin solutions) that are applied after randomization until ICU 
discharge / day 28 whatever occurs first will lead to major PD (no exceptions) 
• 
Albumin that was applied between randomization and 48h after randomization will lead to 
major PD 
• 
Albumin with different indication than solely albumin substitution, albumin supplementation, 
liver failure or hypoalbuminemia from timepoint 48h after randomization until ICU 
discharge/day 28 whatever occurs first will lead to major PD. 
10 Efficacy 
Efficacy analyses will be presented based on ITT set, unless otherwise specified.  
10.1 Primary efficacy analysis 
The primary objective is the verification of the difference in the time to achieve first / initial HDS using 
a gelatin solution combined with a balanced electrolyte solution (gelatin group) versus a balanced 
electrolyte solution (crystalloid group), so that the following hypothesis needs to be tested:  
 
𝐻0: 𝐻𝐷𝑆𝑔𝑒𝑙𝑎𝑡𝑖𝑛= 𝐻𝐷𝑆𝑐𝑟𝑦𝑠𝑡𝑎𝑙𝑙𝑜𝑖𝑑  
𝐻1: 𝐻𝐷𝑆𝑔𝑒𝑙𝑎𝑡𝑖𝑛≠𝐻𝐷𝑆𝑐𝑟𝑦𝑠𝑡𝑎𝑙𝑙𝑜𝑖𝑑 
 
The primary efficacy endpoint is then defined as the time from first administration of IPs to first 
achievement of confirmed HDS (time to HDS at timepoint 0h after confirmation at 4h) in hours. A 
patient is thereby considered as haemodynamically stable, if MAP > 65 mmHg and at least two of the 
following three criteria are fulfilled: 
1. Arterial lactate decrease within the last 6 h > 10% or lactate < 2.4 mmol/l 
2. Urine production > 0.5 ml/kg/h 
3. ScvO2 > 70% 
Once these criteria are fulfilled for the first time, treatment with study fluids temporarily stops and 
the patient is monitored at 2 and 4 h after the first fulfilment of the haemodynamic stability criteria. 
If during these 4 hours the patient remains stable (criteria fulfilment) and there is no need to increase 
inotrope and / or vasopressor therapy and a maximum of 1 l additional study fluids is administered 
for more than 30 min due to sedation, the patient is considered stable and treatment with fluids 
definitively stops. The confirmation of the HDS at the 2h and 4h will be selected using the respective 
timepoints collected in the database. The first 0h time where the criteria for HDS are fulfilled and 
where HDS was confirmed 2 and 4 hours thereafter will be used for the calculation of the primary 
parameter, i.e. time to HDS.  


 
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Patients that do not achieve HDS will be excluded from the primary endpoint analysis. A second 
analysis will be done imputing the missing time to HDS values to the worst-case scenario using the 
worst time to HDS value for the Gelatin group and the best time to HDS value for the Crystalloid group.  
Data related to the HDS will be taken from the eCRF page “Haemodynamic parameters (HDS)”. 
The time to HDS (hours) will be summarized descriptively per treatment group based on the ITT set, 
and the treatment difference will be analyzed by use of a non-parametric test called Mann-Whitney 
U-test. The stratification variables will be considered covariates in a second non-parametric analysis 
of covariance (Van Elteren test). A box plot will also be presented for time to HDS by treatment group, 
in order to observe groups distributions and detect potential outliers. Moreover, first achieved HDS 
parameters will be summarized by treatment group at timepoint 0h, 2h and 4h. 
A data listing presenting the information collected under the eCRF page “Haemodynamic parameters 
(HDS)” will be created.  
10.2 Secondary efficacy analyses 
The development of different parameters is assessed to closely monitor the patient during the acute 
phase in ICU. These parameters are used to investigate efficacy of the IPs by analysing the 
development over time.  
Initial planned treatment comparisons were going to be made at the last recorded value per patient. 
However, during the preparation of the BDRM it revealed that patients discontinued at different 
timepoints. Thus, this analysis is not meaningful from a clinical/statistical perspective. Moreover, the 
team observed a general high quantity of missing data for timepoints after day 6, where less than 50% 
of the patients recorded data for the planned parameters in most of the cases. For example, only 
around 20% of patients in ITT set recorded values of cardiac index during treatment period, and the 
percentage decreases for cardiac index minimum and maximum after day 3.  Therefore, it was agreed 
to remove the treatment comparisons for these secondary endpoints and simply present the 
descriptive summaries by visit in an exploratory manner. 
In case more than one value per patient, parameter, and timepoint is recorded, only the first non-
missing value will be presented in summary tables. All recorded values will be presented in data 
listings. 
Fluid responsiveness 
The following parameters will be summarized by treatment group based on ITT set over the different 
collection timepoints: 
• 
Volume responsiveness upon passive leg raising (PLR) with MAP values and change (%). The 
first non-missing value recorded per patient and timepoint will be used for the summary. 
• 
Volume responsiveness by SVI with SVI values, SVI change (%) and MAP (mmHg). The first 
value non-missing recorded per patient and timepoint will be used for the summary. 
Haemodynamic parameters  
• 
Mean arterial pressure (MAP) [mmHg]  
• 
Stroke volume index (SVI) [ml/m2]  
• 
Cardiac index (CI) [l/min/m2] during treatment period, CI min and CI max for the rest of 
timepoints 


 
 
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• 
Central venous pressure (CVP) [mmHg] during treatment period, CVP min and CVP max for 
the rest of timepoints 
• 
Systolic arterial pressure (SAP) [mmHg] during treatment period, SAP min and SAP max for the 
rest of timepoints 
• 
Diastolic arterial pressure (DAP) [mmHg] during treatment period, DAP min and DAP max for 
the rest of timepoints 
Parameters assessing tissue oxygenation and acid base balance / blood gas analysis (BGA) 
The following parameters will be summarized by treatment group and specimen type (separately and 
all together) including the change from first assessment based on ITT set: 
• 
Partial pressure of carbon dioxide (pCO2) [mmHg] 
• 
Partial pressure of oxygen (pO2) [mmHg] 
• 
Hydrogen carbonate (HCO3) [mmol/l] 
• 
Arterial oxygen saturation (SaO2) [%] 
• 
Haemoglobin (Hb) [g/dl] 
• 
Haematocrit (Hct) [%] 
• 
Pondus hydrogenii (pH) 
• 
Base excess (BE) [mEq/l] 
• 
Sodium (Na+) [mmol/l] 
• 
Potassium (K+) [mmol/l] 
• 
Calcium (Ca2+) [mmol/l] 
• 
Chloride (CI-) [mmol/l] 
• 
Central venous oxygen saturation (ScvO2) [%] 
• 
Lactate [mmol/l] 
• 
Lactate decrease [%] 
• 
Arterial oxygen content (CaO2) [mL/dL] = (1.34×SaO2×Hb) + (0.003×paO2) 
• 
Oxygen delivery (DO2) [ml/min] = 10 × CI × CaO2 
• 
Urine output [mL] 
10.3 Other secondary analyses 
Other secondary variables will be assessed to compare the patient’s status during the treatment 
period. Secondary target variables will also be evaluated with nonparametric tests according to their 
scaling, using a Mann-Whitney U-test or Chi-square test. Whereby, in case of a small random sample 
size or an unbalanced condition, Fisher exact test will be used for the categorical comparisons. The 
assessment of a statistical test will be finally confirmed following the data available at DBL.  


 
  
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If obtained p-values are lower than α=0.05, then the null hypothesis of an equal treatment effect will 
be rejected. 
Parameters assessing clinical outcome 
• 
Length of stay in ICU (days): date of ICU discharge-date of ICU admission+1 
• 
Length of stay in hospital (days): date of hospital discharge-date of hospital admission+1 
• 
Cumulative days on renal replacement therapy (RRT) during study and follow-up, separately: 
Trigger for starting RRT will be also summarized over time during the study presenting 
frequencies for the trigger options and summarizing descriptively the duration of the RRT. 
• 
Number of infection free days  
• 
Number of antibiotic free days  
• 
Number of ventilator free days  
• 
Number of vasopressor free days  
• 
Number of patients that needed mechanical ventilation, and patients with ongoing 
mechanical ventilation 
• 
Cumulative days on invasive mechanical ventilation  
• 
Total number of new severe sepsis/septic shock per patient, type of sepsis and presence of 
infection 
Time from first IP administration to ICU discharge and to ICU discharge during study will be also 
analyzed with a Kaplan-Meier plot and treatment comparison will be done with a log-rank test. 
Patients without ICU discharge fulfilled will be censor to the last time they are known to be in ICU or 
to their planned end of the study date (i.e. up to day 90 plus 14 days), respectively. 
Data listings containing more information about each one of the parameters will be presented.  
Additionally, a summary table for the SOFA score measurements and their change from baseline will 
be presented by treatment group, as well as the corresponding data listing. Similar analyses will be 
presented for SAPS II and APACHE II data that was collected with prior protocol versions. 
Analyses related to the need of RRT therapy will be repeated by the subgroup of SOFA renal score at 
baseline. 
10.4 Exploratory analyses 
Exploratory analyses will be presented to compare the current trial definition of severe sepsis and 
septic shock with the new definitions specified in the article from Mervyn Singer et al, 2016. 
Sepsis is newly defined as life-threatening organ dysfunction caused by a dysregulated host response 
to infection. Organ dysfunction can be identified as an acute change in total SOFA score ≥ 2 points 
consequent to the infection. The baseline SOFA score can be assumed to be zero in patients not known 
to have pre-existing organ dysfunction. 
Patients with septic shock can be identified now with a clinical construct of sepsis with persisting 
hypotension requiring vasopressors to maintain MAP ≥ 65mmHg and having a serum lactate level >2 
mmol/L (18mg/dL) despite adequate volume resuscitation. 


 
 
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A summary table will present the frequencies of patients with new sepsis/septic shock as recorded in 
the database and as per new definitions above.  
11 Safety 
Safety analyses will be presented based on SAF set. 
11.1 Adverse events 
Adverse events (AE) are coded using MedDRA version 24.1 and presented by primary System Organ 
Class (SOC) and Preferred Term (PT).  
The analysis will focus on the treatment-emergent AEs (TEAE), i.e., AEs which started or worsened on 
or after the first study medication intake. Additionally, Adverse Reactions (AR) will be analysed, 
defined as all TEAEs judged by the reporting investigator as having a reasonable causal relationship to 
the IP. If relationship to study drug is missing, the event will be assessed as unrelated if it started 
before start of study medication; in all other cases it will be assumed to be related. If seriousness 
criteria is missing, the event will be considered serious.  
Adverse events of special interest (AESIs) are defined as those whose SOC/PT is one of the following 
list: 
• 
Renal and urinary disorders/Acute kidney injury 
• 
Renal and urinary disorders/Renal failure 
• 
Investigations/Blood creatinine decreased 
• 
Surgical and medical procedures/Renal replacement therapy 
 
Number and frequencies of patients with TEAEs and number of TEAEs will be presented based on SAF 
set in an overall table and given by SOC and by PT within each SOC for the following: 
• 
All TEAEs by treatment group 
• 
Serious TEAEs by treatment group 
• 
Non-serious TEAEs by treatment group 
• 
All ARs by treatment group 
• 
Non-treatment related TEAEs by treatment group 
• 
Serious ARs by treatment group 
• 
Non-serious ARs by treatment group 
• 
Severe TEAEs by treatment group 
• 
Severe ARs by treatment group 
• 
TEAEs of special interest by treatment group 
• 
ARs of special interest by treatment group 
• 
TEAEs leading to death by treatment group (if occurring in more than 5 patients, otherwise a 
listing will be sufficient) 


 
 
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• 
TEAEs leading to study discontinuation by treatment group (if occurring in more than 
5 patients, otherwise a listing will be sufficient) 
• 
ARs leading to death by treatment group (if occurring in more than 5 patients, otherwise a 
listing will be sufficient) 
• 
ARs leading to study discontinuation by treatment group (if occurring in more than 5 patients, 
otherwise a listing will be sufficient) 
Number and frequencies of patients with TEAEs and ARs will be presented by decreasing frequency of 
the overall group. A patient with more than one event in the same SOC or PT will be counted only 
once within the respective SOC and PT, respectively.  
All TEAEs and ARs will be also presented by maximum severity, PT and SOC and by treatment group. 
If severity is missing, the event will not be included in the frequency tables presenting events by 
intensity.  
A frequency table by treatment group of the number of patients that died during study (i.e. before 
ICU discharge or day 28), during follow-up, until follow-up 1, and after follow-up 1 will be presented, 
and the death cause will be also presented. Additionally, a Kaplan-Meier analysis of time to death 
within the planned study period (i.e. up to day 90 plus 14 days) and up to the last date patients are 
known to be alive will be presented separately. Patients that didn’t die during the study will be 
censored to the planned end of study as per study protocol or to the last time they are known to be 
alive, respectively. Additionally, a Cox regression model over time to death will be analyzed using the 
time to HDS, the treatment, the prior RBC therapy as confirmed in the eCRF, and the SOFA renal score 
at baseline as independent variables. 
Moreover, a box plot of APACHE II score at baseline by mortality status will be created. 
In order to assess a potential relationship between HDS and mortality, a logistic regression over the 
mortality status (yes/no) will be presented, using the HDS (yes/no), the treatment, the prior RBC 
therapy as confirmed in the eCRF, and the baseline SOFA renal score as independent variables 
Separated data listings for all AEs and all serious AEs will be presented, indicating if the event is an AR. 
11.2 Vital signs 
Vital signs parameters are assessed within APACHE II parameters (section 7.2) and as part of secondary 
endpoints (haemodynamic parameters) in section 7.2 and will therefore not be analyzed separately. 
Only temperature measurements (overall, minimum, and maximum) taken during the treatment 
course will be summarized additionally. 
11.3 Clinical safety laboratory  
Summary tables over time by treatment group and box plots will be provided for all haematology and 
clinical chemistry parameters that were not analyzed as secondary variables (see section 10.2), 
including the change from the baseline or first assessment value. They are:  
• 
Coagulation parameters: prothrombin time (PT), activated partial thromboplastin time (aPTT), 
international norm ratio (INR), and site-specific parameters (Innsbruck): fibrinogen, 
antithrombin (AT), and platelets absolute. 
• 
Hepatic function parameters: total bilirubin 


 
 
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• 
Renal function parameters: serum creatinine (SCr), serum blood urea nitrogen (BUN), 
Procalcitonin, and: 
o Creatinine clearance (Ccr) using Cockcroft-Gault-Formula 
𝐶𝑐𝑟= (140 −𝑎𝑔𝑒)𝑥 𝑤𝑒𝑖𝑔ℎ𝑡 [𝑘𝑔]
72 𝑥 𝑆𝐶𝑟[𝑚𝑔/𝑑𝑙]
 𝑥 (0.85, 𝑖𝑓 𝑓𝑒𝑚𝑎𝑙𝑒) 
o SCr-based eGFR using CKD-EPI (SCr)-equation (Inker et al., 2012) until day 7 
 
 
 
 
 
 
 
Serum creatinine summary and box plot will be repeated for the renal SOFA subgroup defined in 
section 5.4. 
Additionally, serum creatinine and urine output (randomization until 48h) measured during the 
treatment period will be summarized by timepoint and repeated for the subgroups defined in section 
5.4. 
11.4 Kidney Disease Improving Global Outcome (KDIGO) score 
The KDIGO score calculated as specified in appendix 18.4 will be used as basis for the summary 
analyses. A shift table with the KDIGO stages during the study period (until day 7) will be presented 
based on the table specified in appendix 18.4. 
KDIGO score values calculated and also the ones collected in the eCRF will be listed. 
12 Sensitivity Analyses 
Primary endpoint will be also analyzed throughout a survival analysis with Kaplan-Meier estimates and 
a stratified log-rank test. Patients that do not achieve HDS will be censored to the last HDS assessment 
date where they were free of the event of interest. 
Primary and secondary endpoints will be repeated on the PP set as specified in section 4, as well as 
demographics and main baseline characteristics outputs and safety analyses. 
Additionally, as protocol version 6.0 changed some inclusion/exclusion criteria, primary endpoint 
analysis for patients that were enrolled with prior protocol versions vs. patients that were enrolled 
after protocol version 6.0 will be presented. 
No outliers are identified in the primary endpoint analysis during the preparation of the BDRM, 
therefore no further sensitivity analyses without the detected outliers will be presented as needed. 
Sex 
SCr 
Formula 
Female ≤ 0.7mg/dl  144 x (SCr/0.7)-0.329  x 0.993Age  [x 1.159 if black] 
Female > 0.7mg/dl  144 x (SCr/0.7)-1.209  x 0.993Age  [x 1.159 if black] 
Male 
≤ 0.9mg/dl 141 x (SCr/0.9)-0.411  x 0.993Age  [x 1.159 if black] 
Male 
> 0.9mg/dl 141 x (SCr/0.9)-1.209  x 0.993Age  [x 1.159 if black] 


 
  
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Subgroups analyses on primary endpoint as specified in section 5.4 will be also presented reproducing 
the analyses planned for this endpoint based on the different subgroup categories. Similar analyses 
by subgroups will be done for TEAEs, serious TEAEs, mortality and serum creatinine values during 
treatment period (from randomization until 48h). 
Further sensitivity analyses may be defined during the course of the study as applicable. 
13 Follow-up analyses 
The following variables will be analyzed as part of the follow-up period based on ITT set. 
13.1 Follow-up therapies 
Data listings and summary statistics will be presented containing information about the follow-up 
data collected at day 28 and day 90 for: 
• 
Mortality including cause of death, if applicable  
• 
Colloid therapy administered (only day 28) 
• 
Last available serum creatinine data (only day 28) 
• 
Renal replacement therapy (RRT) or new kidney disease including cause, if applicable (only 
day 90) 
13.2 Health Related Quality of Life (HRQoL) 90 days after randomization 
Patients’ quality of life will be measured by using the HRQoL questionnaire (EQ-5D-5L, EuroQol Group). 
Frequencies for each one of the subscales (mobility, self-care, usual activities, pain/discomfort, 
anxiety/depression) will be presented together with the summary statistics for the total health score 
(ranging from 0 to 100). The number of missing responses will also be displayed. Results of the 
questionnaire will be also listed. 
Each dimension has 5 levels (no problems, slight problems, moderate problems, severe problems, and 
extreme problems). The respondent is asked to indicate his/her health state or placing a cross in the 
box against the most appropriate level within each of the 5 dimensions. This decision results in a 1-
digit number expressing the level selected for that dimension. The user guide emphasises that the 
numerals 1-5 for each level have no arithmetic properties and should not be used as a cardinal score. 
Ambiguous item values (e.g., 2 levels are ticked for a single dimension) will be treated as missing 
values. 
14 Interim Analysis 
An interim analysis was planned after inclusion of 400 patients to perform a recalculation of the 
sample size. 
However, the DSMB decided to prematurely terminate the study after inclusion of 167 patients, on 
8th of December of 2021. Therefore, the interim analysis will not be performed. 
15 Statistical Analyses for Safety Monitoring 
A DSMB was appointed for this study. Further responsibilities and tasks were defined separately in a 
DSMB charter. 


 
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16 Software  
If not stated otherwise, the data will be analyzed using SAS version 9.4.  
 


 
 
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17 Abbreviations 
AE 
Adverse Event 
ANCOVA 
Analysis of COVARIANCE 
APACHE II 
Acute Physiology And Chronic Health Evaluation II 
aPTT 
Activated partial thromboplastin time 
AR 
Adverse reaction 
AT 
Antithrombin 
BDRM 
Blind data review meeting 
BE 
Base excess 
BGA 
Blood gas analysis 
BMI 
Body mass index 
BSA 
Body surface area 
BUN 
Blood urea nitrogen 
Ca2+ 
Calcium 
CaO2 
Arterial oxygen content 
Ccr 
Creatinine clearance 
CI 
Cardiac Index 
CL- 
Chloride 
CO2 
Carbon dioxide 
CP 
Conditional power 
eCRF 
Electronic Case Report Form 
CVP 
Central venous pressure 
DAP 
Diastolic arterial blood pressure 
DBL 
Database lock 
DO2 
Oxygen delivery 
DSMB 
Data Safety Monitoring Board 
eGFR 
Estimated glomerular filtration rate 
FC 
Fluid challenge 
Hb 
Haemoglobin 
HCO3 
Hydrogen carbonate 
Hct 
Haematocrit 
HDS 
Haemodynamic stability 
HES 
Hydroxyethyl starch 
HRQoL 
Health related quality of life 
IA 
Interim Analysis 
ICU 
Intensive care unit 
INR 
International norm ratio 
IP 
Investigational product 
ITT 
Intention to Treat 
K+ 
Potassium 


 
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KDIGO 
Kidney Disease Improving Global Outcome 
MANOVA 
Multivariate analysis of variance 
MAP 
Mean arterial pressure 
MedDRA 
Medical Dictionary for Regulatory Activities 
Na+ 
Sodium 
pCO2 
Partial pressure of carbon dioxide 
pH 
Pondus hydrogenii 
pO2 
Partial pressure of oxygen 
PLR 
Passive leg raising 
PP 
Per Protocol 
PT 
Preferred Term 
QoL 
Quality of Life 
RBC 
Red blood cell 
RRT 
Renal replacement therapy 
SAF 
Safety analysis set 
SaO2 
Arterial oxygen saturation 
SAP 
Statistical Analysis Plan 
SAP 
Systolic arterial blood pressure 
SAPS II 
Simplified Acute Physiology Score 
SBP 
Systolic blood pressure 
SCr 
Serum creatinine 
ScvO2 
Central venous oxygen saturation 
SOC 
System Organ Class 
SOFA 
Sequential Organ Failure Assessment-score 
SSR 
Sample size recalculation 
SVI 
Stroke volume index 
TEAE 
Treatment Emergent Adverse Event 
TLF 
Tables, Listings, and Figures 


 
 
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18 Appendices 
18.1 APACHE II score 
The APACHE II score will be calculated using the following criteria: 
Variable 
+4 
+3 
+2 
+1 
0 
+1 
+2 
+3 
+4 
Temperature (ºC) 
>=41 
39-
40.9 
 
38.5-
38.9 
36-
38.40 
34-
35.9 
32-
33.9 
30-
31.9 
<=29.9 
Mean arterial pressure (MAP) [mmHg] 
>=160 
130-
159 
110-
129 
 
70-
109 
 
50-
69 
 
<=49 
Heart rate (bpm)  
>=180 
140-
179 
110-
139 
 
70-
109 
 
55-
69 
40-
54 
<=39 
Respiratory rate (bpm) 
>=50 
35-
49 
 
25-
34 
12-
24 
10-
11 
6-9 
 
>=5 
Oxygenation [mmHg] (if FiO2 >=0.5 use AaDO2, if 
FiO2<0.5 use PaO2) 
>=500 
350-
499 
200-
349 
 
<200 
or 
>70 
PaO2 
61-
70 
PaO2 
 
55-
60 
PaO2 
<55 
PaO2 
Arterial PH 
>=7.7 
7.6-
7.69 
 
7.5-
7.59 
7.33-
7.49 
 
7.25-
7.32 
7.15-
7.24 
<7.15 
Serum Sodium [mmol/L] 
>=180 
160-
179 
155-
159 
150-
154 
130-
149 
 
120-
129 
111-
119 
<=110 
Potassium [mmol/L] 
>=7 
6-
6.9 
 
5.5-
5.9 
3.5-
5.4 
3-3.4 
2.5-
2.9 
 
<2.5 
Serum Creatinine [mg/dL] 
Double point score for acute renal failure 
>=3.5 
2-
3.4 
1.5-
1.9 
 
0.6-
1.4 
 
<0.6 
 
 
Haematocrit [%] 
>=60 
 
50-
59.9 
46-
49.9 
30-
45.9 
 
20-
29.9 
 
<20 
White blood count [x10^3/mm3] 
>=40 
 
20-
29.9 
15-
19.9 
3-
14.9 
 
1-2.9 
 
<1 
 
Variable 
0 
2 
3 
5 
6 
Age (years) 
<=44 
45-54 
55-64 
65-74 
>=75 
 
Variable 
5 
2 
Chronic health points 
Nonoperative 
or emergency 
postoperative 
patient 
Elective 
postoperative 
patient 
 
Glasgow coma score=15-recorded score 
 
The total APACHE II score is then calculated summing the points as specified in the tables above for 
each variable plus the Glasgow coma score. 
If at least one variable is missing, the total APACHE II score will be missing, except for the chronic 
health points result that is not mandatory to fill for all patients. 
18.2 SOFA score 
The SOFA score is calculated using a range of points 0-4 among the distinct items: 
Criteria 
0 
+1 
+2 
+3 
+4 
Respiratory system 
  PaO2/FiO2 [mmHg] 
≥400 
<400 
<300 
<200 with 
respiratory 
support 
<100 with 
respiratory 
support 
Coagulation 
  Platelets×103/μl 
≥150 
<150 
<100 
<50 
<20 
Liver 
<1,2 
1,2–1,9 
2,0–5,9 
6,0–11,9 
>12,0 


 
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  Bilirubin (mg/dl)  
Cardiovascular system 
Mean arterial pressure OR 
administration of 
vasopressors required 
MAP ≥70 mmHg 
MAP <70 mmHg 
dopamine ≤ 5 
μg/kg/min or 
dobutamine (any 
dose) 
dopamine 5,1-15 
μg/kg/min OR 
epinephrine ≤ 
0.1 μg/kg/min 
OR 
norepinephrine ≤ 
0.1 μg/kg/min 
dopamine > 15 
μg/kg/min OR 
epinephrine > 0.1 
μg/kg/min OR 
norepinephrine > 
0.1 μg/kg/min 
Nervous system 
  Glasgow coma scale 
15 
13–14 
10–12 
6–9 
<6 
Kidneys 
  Creatinine (mg/dl) 
<1,2 
1,2–1,9 
2,0–3,4 
3,5–4,9 
>5,0 
  Urine output (mL/d) 
 
 
 
<500 
<200 
 
The total SOFA score is calculated by summing up the scores obtained for each one of the categories. 
Higher values are related to worse patient status. 
If at least one criterion is missing, the total SOFA score will be missing. 
18.3 SAPS II score 
Criteria 
26 
13 
12 
11 
9 
7 
6 
5 
4 
3 
2 
0 
Age, y 
 
 
 
 
 
 
 
 
 
 
 
<40 
Heart rate, beats/min 
 
 
 
<40 
 
 
 
 
 
 
40-
69 
70-119 
Systolic BP, mmHg 
 
<70 
 
 
 
 
 
70-
99 
 
 
 
100-199 
Body temperature, C 
 
 
 
 
 
 
 
 
 
 
 
<39 
Only if ventilated or continuous 
pulmonary artery pressure Pao2, mm 
Hg/Fio2 
 
 
 
<100 
100-
199 
 
>=200 
 
 
 
 
 
PaOz, kPa/Fio2 
 
 
 
<13.3 
13.3-
26.5 
 
>=26.6 
 
 
 
 
 
Urinary output, LVd 
 
 
 
<0.500 
 
 
 
 
0.500-
0.999 
 
 
>=1000 
Serum urea level, mmol/L or 
serum urea nitrogen level, mg/dL 
 
 
 
 
 
 
 
 
 
 
 
<10.0 
<28 
WBC count (107cu mm) 
 
 
<1.0 
 
 
 
 
 
 
 
 
1.0-19.9 
Serum potassium, mmol/d 
 
 
 
 
 
 
 
 
 
<3.0 
 
3.0-4.9 
Serum sodium level, mmol/L 
 
 
 
 
 
 
 
<125 
 
 
 
125-144 
Serum bicarbonate level, mEq/L 
 
 
 
 
 
 
<15 
 
 
15-
19 
 
>=20 
Bilirubin level, μ /L  
 
 
 
 
 
 
 
 
 
 
 
<68.4 
Glasgow Coma Score 
<6 
6-8 
 
 
 
9-
10 
 
11-
13 
 
 
 
14-15 
Chronic diseases 
 
 
 
 
 
 
 
 
 
 
 
 
Type of admission 
 
 
 
 
 
 
 
 
 
 
 
Scheduled 
surgical 
 
Criteria 
1 
2 
3 
4 
6 
7 
8 
9 
10 
12 
15 
17 
18 
Age, y 
 
 
 
 
 
40-59 
 
 
 
60-
69 
70-
74 
 
>=80 
Heart rate, 
beats/min 
 
 
 
120-
159 
 
>=160 
 
 
 
 
 
 
 
Systolic BP, 
mmHg 
 
>=200 
 
 
 
 
 
 
 
 
 
 
 
Body 
temperature, 
C 
 
 
>=39 
 
 
 
 
 
 
 
 
 
 
Only if 
ventilated or 
 
 
 
 
 
 
 
 
 
 
 
 
 


 
  
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continuous 
pulmonary 
artery 
pressure 
Pao2, mm 
Hg/Fio2 
PaOz, 
kPa/Fio2 
 
 
 
 
 
 
 
 
 
 
 
 
 
Urinary 
output, LVd 
 
 
 
 
 
 
 
 
 
 
 
 
 
Serum urea 
level, 
mmol/L  or 
serum urea 
nitrogen 
level, mg/dL 
 
 
 
 
10.0-
29.9 
28-83 
 
 
 
>=30 
>=84 
 
 
 
 
WBC count 
(107cu mm) 
 
 
>=20 
 
 
 
 
 
 
 
 
 
 
Serum 
potassium, 
mmol/d 
 
 
>=5.0 
 
 
 
 
 
 
 
 
 
 
Serum 
sodium level, 
mmol/L 
>=145 
 
 
 
 
 
 
 
 
 
 
 
 
Serum 
bicarbonate 
level, mEq/L 
 
 
 
 
 
 
 
 
 
 
 
 
 
Bilirubin 
level, μ /L 
 
 
 
68.4-
102.5 
 
 
 
>=102.6 
 
 
 
 
 
Glasgow 
Coma Score 
 
 
 
 
 
 
 
 
 
 
 
 
 
Chronic 
diseases 
 
 
 
 
 
 
 
Metastatic 
cancer 
Hematologic 
malignancy 
 
 
AIDS 
 
Type of 
admission 
 
 
 
 
Medical 
 
Unscheduled 
surgical 
 
 
 
 
 
 
 
The SAPS II score is made up of 17 variables: 12 physiological variables, age, type of admission 
(scheduled surgical, unscheduled surgical, or medical), and three variables related to underlying 
disease: acquired immunodeficiency syndrome, metastatic cancer, and hematologic malignancy. 
Points assigned for each variable vary from 0 to 3 (for temperature) up to 0 to 26 (for Glasgow Coma 
Score).  
For the 12 physiological variables, the worst value during the first 24 hours in the ICU is taken into 
account.  
No arterial sample is necessary if the patient is not ventilated or receiving continuous positive airway 
pressure.  
For sedated patients, the Glasgow Coma Score before sedation was used. This was ascertained either 
from interviewing the physician who ordered the sedation, or by reviewing the patient's medical 
record.  
The total SAPS score is calculated by summing up the scores obtained for each one of the categories. 
Higher values are related to worse patient status.  
If at least one criterion is missing, the total SAPS II score will be missing. 
18.4 KDIGO score 
Stages will be derived based on the following values of serum creatinine: 
 


 
 
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Stage 
Serum creatinine 
1 
1.5–1.9 times baseline 
OR 
≥ 0.3 mg/dl (≥ 26.5 mmol/l) increase 
2 
2.0–2.9 times baseline 
3 
3.0 times baseline 
OR 
Increase in serum creatinine to ≥ 4.0 mg/dl (≥ 
353.6 mmol/l) 
OR 
Initiation of renal replacement therapy 
OR 
In patients < 18 years, decrease in eGFR to < 35 
ml/min per 1.73 m2 
 
For calculating baseline KDIGO scores, the simplified 'modification of diet in renal disease' (MDRD) 
formula provides a robust estimate of GFR relative to serum creatinine based on age, race and sex.  
Therefore, the recorded serum creatinine value at baseline visit will be compared using the KDIGO 
scoring table with the estimated value following this formula. The following protocol table establishes 
the estimated baseline creatinine that will be used for comparison using: 
Age (years)  
Black males  
mg/dl (umol/l)  
Other males  
mg/dL (umol/l)  
Black females  
mg/dl (umol/l)  
Other females  
mg/dl (umol/l)  
20-24  
1.5 (133)  
1.3 (115)  
1.2 (106)  
1.0 (88)  
25-29  
1.5 (133)  
1.2 (106)  
1.1 (97)  
1.0 (88)  
30-39  
1.4 (124)  
1.2 (106)  
1.1 (97)  
0.9 (80)  
40-54  
1.3 (115)  
1.1 (97)  
1.0 (88)  
0.9 (80)  
55-65  
1.3 (115)  
1.1 (97)  
1.0 (88)  
0.8 (71)  
>65  
1.2 (106)  
1.0 (88)  
0.9 (80)  
0.8 (71)  
 
 
 
 


 
  
Version: 2.0 
Version Date: 05JUL2022 
Statistical Analysis Plan: final analysis  
 
 
QMS Document Name: Statistical Analysis Plan 
Page 33 of 33 
 
Document date: 09JUL2019 
CONFIDENTIAL 
 
19 References 
1. ICH Harmonised Tripartite Guideline E9, Statistical Principles for Clinical Trials, 1998 
2. ICH Harmonised Tripartite Guideline E6(R1), Guideline for Good Clinical Practice, 1996 
3. Adaptive Designs for Clinical Trials of Drugs and Biologics, Guidance for Industry, FDA, 2019 
4. Vincent JL, Moreno R, Takala J, Willatts S, De Mendonça A, Bruining H, Reinhart CK, Suter PM, Thijs 
LG (Jul 1996). "The SOFA (Sepsis-related Organ Failure Assessment) score to describe organ 
dysfunction/failure. On behalf of the Working Group on Sepsis-Related Problems of the European 
Society of Intensive Care Medicine". Intensive Care Med. 22 (7): 707–10. doi:10.1007/bf01709751. 
PMID 8844239. 
5. Knaus WA, et al. APACHE II: a severity of disease classification system. Crit Care Med. 1985 
Oct;13(10):818-29. PubMed ID: 3928249  
6. Guidet, Bertrand, et al. "Assessment of hemodynamic efficacy and safety of 6% 
hydroxyethylstarch 130/0.4 vs. 0.9% NaCl fluid replacement in patients with severe sepsis: The 
CRYSTMAS study." Crit Care 16.3 (2012): R94. 
 
7. Inker, Lesley A., et al. "Estimating glomerular filtration rate from serum creatinine and cystatin 
C." New England Journal of Medicine 367.1 (2012): 20-29. 
 
8. Singer, Mervyn et al. “The Third International Consensus Definitions for Sepsis and Septic Shock 
(Sepsis-3).” JAMA vol. 315,8 (2016): 801-10. doi:10.1001/jama.2016.0287 
 
9. BBraun_TLFs shells Final 2.0