Defibrotide in the Human Endotoxemia Model --- an Exploratory Trial Investigating the Effects and the Mechanisms of Defibrotide

Study Overview
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Brief Summary
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Defibrotide is an anti-inflammatory and anti-coagulatory agent approved for treatment of veno-occlusive disease. Although it has been in clinical use for almost 30 years, the exact mechanism of action has never been fully elucidated. Thus, the effects of defibrotide will be investigated in the human endotoxemia model in order to gather further information on its actions.

Detailed Description
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Defibrotide (DF) is a highly complex polydisperse mixture of single-stranded phosphodiester oligodeoxyribonucleotides derived from the controlled depolymerization of porcine intestinal mucosal DNA. The entire mode of action remains unknown. Its actions may be summarized to pro-fibrinolytic, anti-inflammatory and anti-coagulatory actions. To better define the mechanisms of Defibrotide the effects of the substance will be investigated in the well-established endotoxemia model. Sixteen healthy volunteers will be randomized to receive LPS±defibrotide/placebo and four subjects will be randomized to receive Placebo± defibrotide/placebo in a single center, randomized, double blind, placebo controlled, two-way crossover trial. Immediately after a 2h infusion of 6,25mg/kg bodyweight defibrotide or placebo a LPS bolus of 2ng/kg bodyweight will be infused. Analyses will be performed by blood sampling at pre-defined time-points.

Official Title
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Defibrotide in the Human Endotoxemia Model -- an Exploratory Trial Investigating the Effects and the Mechanisms of Defibrotide

Conditions
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Healthy Volunteers, Endotoxemia

Intervention / Treatment
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* Drug: Defibrotide
* Drug: Placebo (0.9% sodium chloride)
* Drug: Lipopolysaccharide
* Drug: Placebo (0.9% sodium chloride bolus)


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: >18 years of age <90kg body weight Normal findings in medical history and physical examination unless the investigator considers the abnormality to be clinically irrelevant Normal laboratory values unless the investigator considers abnormalities to be clinically irrelevant Ability to understand the purpose and nature of the study, as well as the associated risks Exclusion Criteria: Intake of any drugs that may interfere with the trial's endpoints or drugs (i.e. platelet inhibitors, anticoagulants, etc.) Positive results of HIV or hepatitis virology Acute illness with systemic inflammatory reactions Known allergies, hypersensitivities or intolerances to any of the used substances Acute or recent bleeding episodes, increased risk of bleeding at the discretion of the investigator Participation in an LPS trial within 6 weeks of the first study day Pregnancy or breastfeeding

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Crossover Assignment
Interventional Model Description: Subjects will be randomized to receive LPS (n=16) or placebo (n=4) first. In each group they will undergo two study periods (crossover trial): a placebo period and a defibrotide period. The placebo group (n=4) will only be analyzed descriptively.
Masking: Quadruple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: Defibrotide/LPS<br>2ng/kg lipopolysaccharide period I: 6.25mg/kg bodyweight defibrotide or placebo (0.9% sodium chloride) period II: vice versa 16 healthy volunteers will receive 2ng/kg bodyweight LPS plus Defibrotide or Placebo 4 healthy volunteers will receive 0.9% saline (NO LPS) plus Defibrotide or Placebo | Drug: Defibrotide<br>* 6.25mg/kg bodyweight over 2h infusion<br>Drug: Lipopolysaccharide<br>* bolus infusion of 2ng/kg bodyweight lps<br>|
| Placebo Comparator: Placebo/LPS<br>2ng/kg lipopolysaccharide period I: 6.25mg/kg bodyweight defibrotide or placebo (0.9% sodium chloride) period II: vice versa 16 healthy volunteers will receive 2ng/kg bodyweight LPS plus Defibrotide or Placebo 4 healthy volunteers will receive 0.9% saline (NO LPS) plus Defibrotide or Placebo | Drug: Placebo (0.9% sodium chloride)<br>* (0.9% sodium chloride) infusion over 2h infusion<br>Drug: Lipopolysaccharide<br>* bolus infusion of 2ng/kg bodyweight lps<br>|
| Other: Defibrotide/Placebo<br>Placebo (0.9% sodium chloride) period I: 6.25mg/kg bodyweight defibrotide or placebo (0.9% sodium chloride) period II: vice versa 16 healthy volunteers will receive 2ng/kg bodyweight LPS plus Defibrotide or Placebo 4 healthy volunteers will receive 0.9% saline (NO LPS) plus Defibrotide or Placebo | Drug: Defibrotide<br>* 6.25mg/kg bodyweight over 2h infusion<br>Drug: Placebo (0.9% sodium chloride bolus)<br>* (0.9% sodium chloride) bolus infusion<br>|
| Other: Placebo/Placebo<br>Placebo (0.9% sodium chloride) period I: 6.25mg/kg bodyweight defibrotide or placebo (0.9% sodium chloride) period II: vice versa 16 healthy volunteers will receive 2ng/kg bodyweight LPS plus Defibrotide or Placebo 4 healthy volunteers will receive 0.9% saline (NO LPS) plus Defibrotide or Placebo | Drug: Placebo (0.9% sodium chloride)<br>* (0.9% sodium chloride) infusion over 2h infusion<br>Drug: Placebo (0.9% sodium chloride bolus)<br>* (0.9% sodium chloride) bolus infusion<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Prothrombin Fragments f1+2 | Based on the changes from baseline an area-under the concentration-time curve will be calculated and will be compared between the placebo and the verum phase within each subject. The respective arbitrary unit therefore is fold*h. The placebo period (4 subjects who did not receive lipopolysaccharide) was only included for a descriptive comparison, but not for statistical comparison | The parameter was assessed at baseline, at 0h, 1h, 2h, 4h, 6h, 24h. | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Thrombin-Antithrombin Complexes | Based on the changes from baseline an area-under the concentration-time curve will be calculated and will be compared between the placebo and the verum phase within each subject. The respective arbitrary unit therefore is fold*h. The placebo period (4 subjects who did not receive lipopolysaccharide) was only included for a descriptive comparison, but not for statistical comparison | This parameter was assessed at baseline, at 0h, 1h, 2h, 4h, 6h, 24h and AUC was calculated based on these measurements. | 
| Plasmin-Antiplasmin Complexes | Based on the changes from baseline an area-under the concentration-time curve will be calculated and will be compared between the placebo and the verum phase within each subject. The respective arbitrary unit therefore is fold*h. The placebo period (4 subjects who did not receive lipopolysaccharide) was only included for a descriptive comparison, but not for statistical comparison | This parameter was assessed at baseline, at 0h, 1h, 2h, 4h, and 6h and AUC was calculated based on these measurements. | 
| Tumor Necrosis Factor (TNF)-Alpha | Based on the changes from baseline an area-under the concentration-time curve will be calculated and will be compared between the placebo and the verum phase within each subject. The respective arbitrary unit therefore is fold*h. | This parameter was assessed at baseline, at 0h, 1h, 2h, 4h, 6h, 24h and AUC was calculated based on these measurements. | 
| Tissue-type Plasminogen Activator | Based on the changes from baseline an area-under the concentration-time curve will be calculated and will be compared between the placebo and the verum phase within each subject. The respective arbitrary unit therefore is fold*h. | This parameter was assessed at baseline, at 0h, 1h, 2h, 4h, 6h, 24h and AUC was calculated based on these measurements. | 
| Interleukin-6 | Based on the changes from baseline an area-under the concentration-time curve will be calculated and will be compared between the placebo and the verum phase within each subject. The respective arbitrary unit therefore is fold*h. | This parameter was assessed at baseline, at 0h, 1h, 2h, 4h, 6h, 24h and AUC was calculated based on these measurements. | 
| E-Selectin | Based on the changes from baseline an area-under the concentration-time curve will be calculated and will be compared between the placebo and the verum phase within each subject. The respective arbitrary unit therefore is fold*h. | This parameter was assessed at baseline, at 0h, 1h, 2h, 4h, 6h, 24h and AUC was calculated based on these measurements. | 
| Plasminogen Activator Inhibitor 1 | Based on the changes from baseline an area-under the concentration-time curve will be calculated and will be compared between the placebo and the verum phase within each subject. The respective arbitrary unit therefore is fold*h. | This parameter was assessed at baseline, at 0h, 1h, 2h, 4h, 6h, 24h and AUC was calculated based on these measurements. | 
| Von Willebrand Factor Antigen | Based on the changes from baseline an area-under the concentration-time curve will be calculated and will be compared between the placebo and the verum phase within each subject. The quantification of von Willebrand Factor is based on reference values and results are in % of normal. The respective arbitrary unit therefore is %*h. | This parameter was assessed at baseline, at 0h, 1h, 2h, 4h, 6h, 24h and AUC was calculated based on these measurements. | 
| Clotting Time in Thromboelastometry | In this analysis, first of all a ratio of the measurement time point to the baseline was calculated. Thereafter deltas (baeline-ratio) were calculated. With the results an AUC was calculated. The respective arbitrary unit therefore is fold*h. | This parameter was assessed at baseline, at 0h, 1h, 2h, 4h, 6h, 24h and AUC was calculated based on these measurements. | 
| Maximum Lysis in Thromboelastometry | Based on the changes from baseline an area-under the concentration-time curve will be calculated and will be compared between the placebo and the verum phase within each subject. The respective arbitrary unit therefore is fold*h. | This parameter was assessed at baseline, at 0h, 1h, 2h, 4h, 6h and AUC was calculated based on these measurements. | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Coagulation, Inflammation, Defibrotide, Fibrinolysis