The Effects of DLBS1033 on Haemostasis Parameters in Healthy Volunteers

Study Overview
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Brief Summary
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DLBS1033 is bioactive protein fraction which extracted from Lumbricus rubellus earthworm. This earthworm comes from Pengalengan, West Java, Indonesia. DLBS1033 possesses 8 major proteins with molecular weight below 100 kDa, so its named as Lumbricus Low Molecular weight Proteins (LLP). This enzym can be transported to the bloodstream via intestinal epitel. Structure of DLBS1033 looks like lumbrokinase. Lumbrokinase is enzym that consist of 6 isoenzyme serine protease. As a drug that consists of serin protease enzym, suspected that the mechanism of action of DLBS1033 similar with lumbrokinase, especially as plasminogen activator in fibrinolytic system. In vitro study by Trisina et al showed that DLBS1033 has fibrinogenolytic activities on fibrinogen α, beta, and gamma chain, decreasing platelet aggregation and clotting time was prolonged. Until now, the mechanism of action and effects of DLBS1033 on human fibrinolytic and coagulation system still unknown. Therefore, the aim of this clinical trial is to evaluate the effects of DLBS1033 on human fibrinolytic and coagulation system on healthy subjects.

Detailed Description
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Fibrinolysis is a process to lyse clot formed by thrombin. It starts with activating plasminogen to plasmin by the endogenous tissue plasminogen activator (tPA) and urokinase plasminogen activator (uPA). These activator are found in the endothelium, granulocytes and monocytes. Plasminogen activator inhibitor-1 (PAI-1) is the major inhibitor of tPA and uPA) and α2-antiplasmin is major inhibitor of plasmin. Plasmin wil degrade soluble fibrinogen and fibrin to produce fibrinogen degradation products and fibrin degradation products, respectively. Lumbrokinase has been investigated in animal studies. Kim et al was conducting on rats by administered freeze-dried powder of Lumbricus rubellus earthworm orally. This study showed that earthworm powder is valuable for the prevention and/or treatment of thrombotic conditions. Similar conclusion also shown on study by Lee et al, which compared antithrombotic and fibrinolytic activities of lumbrokinase SPP-501 (extracted from Eisenia andrei earthworm) with urokinase and t-PA in a thrombosis model of rat vena. The results of this study were decreasing of thrombus weight, shortening of euglobulin lysis time (ELT), and reducing platelet aggregation of rat which given SPP-501. Conclusion from several clinical trial similar with several studies on animal. From study which was conducted by Jin et al on 51 cerebral infarct subjects which were given 3 times 400 mg lumbrokinase (n = 31) or control (n = 20) for 28 days. In the treatment group, kaolin partial thromboplastin time (KPTT) was prolonged, t-PA activity and D-dimer level increase, and fibrinogen decreased significantly. It is concluded that mechanism of lumbrokinase as oral antithrombotic and fibrinolytic are by inhibition of coagulation intrinsic pathway and activate fibrinolytic pathway by increasing t-PA activity. Fibrinolytic activity was also concluded by Rey, who was conducted study on 28 diabetic foot ulcer subjects, which were given 3 times 500 mg lumbrokinase per day (n = 14) for 7 days, or placebo. On this study, in the treatment group mean of D-dimer was increased. From many clinical trials, it is concluded that effects of lumbrokinase have seen after several days. It is different with intravenous fibrinolytic enzym, such as streptokinase and t-PA, which effects have seen soon after it was used. Therefore oral lumbrokinase could not replace the function of intravenous fibrinolytic enzym,that was used on acute thrombosis. Based on antithrombotic and fibrinolytic activity, lumbrokinase might used as secondary prevention after acute thrombosis, such as myocard infarct and stroke. Some researchers have started clinical trials about that hypothesis. Pilot study by Kasim et al was used a lumbrokinase in 10 patient with stable angina pectoris. This study showed that 70% of total sample receiving lumbrokinase had a significant decrease in summed stress score of perfusion imaging and better perfusion in viable myocardium after 30 days of lumbrokinase treatment.

Official Title
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The Effects of DLBS1033 on Haemostasis Parameters in Healthy Volunteers

Conditions
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Healthy

Intervention / Treatment
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* Drug: DLBS1033
* Drug: placebo


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Male 18-55 years old body mass index between 18-25 kg/square metres normal physical examination Patient still have the ability to undergo examinations and give written informed consent Plasmin-antiplasmin complex (PAP complex) level between 0-514 ng/ml Platelet aggregation (ADP 10 uM) > 49% Exclusion Criteria: Patient with cardiovascular disease, hypertension, diabetes mellitus, and dyslipidemia Creatinin serum more than 1,5 x upper limit normal SGOT and SGPT more than 3 x upper limit normal Blood pressure ≥ 140/90 mmHg Fasting blood glucose > 126 mg/dL Alcohol patients Took any medications (including traditional medicine, supplement and vitamin) in 1 week before the study) Patient has bleeding history which unclear etiology Hemoglobin < 10 g/dL Thrombocyte count < 100.000/uL Heavy smoker (Bringman Index > 600)

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 55 Years

Sexes Eligible for Study
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Male

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: Treatment
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Placebo Comparator: placebo<br>The comparison drug is placebo, which is made with same size and shape with study drug. DLBS1033 and placebo is produced by PT Dexa Medica. Placebo will taken 3 times 1 tablet per day for 7 days. Wash out period before enter another arm is 7 days. | Drug: DLBS1033<br>* The study drug is enteric coated DLBS1033, which contain 490 mg bioactive protein fraction.<br>Drug: placebo<br>* Placebo is made with same size and shape with study drug.<br>|
| Experimental: DLBS1033<br>The study drug is enteric coated DLBS1033, which contain 490 mg bioactive protein fraction. The drug will taken 3 times 490 mg per day for 7 days. Wash out period before enter another arm is 7 days. | Drug: DLBS1033<br>* The study drug is enteric coated DLBS1033, which contain 490 mg bioactive protein fraction.<br>Drug: placebo<br>* Placebo is made with same size and shape with study drug.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| plasmin-antiplasmin complex (PAP complex) | changes in plasmin-antiplasmin complex | one week | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| euglobulin clot lysis time (ECLT) | changes in euglobulin clot lysis time | one week | 
| prothrombin time (PT) | changes in prothrombin time | one week | 
| fibrinogen | changes in fibrinogen | one week | 
| activated partial thromboplastin time (aPTT) | changes in activated partial thromboplastin time | one week | 
| platelet aggregation | changes in platelet aggregation | one week | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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lumbrokinase, fibrinolytic, plasmin-antiplasmin complex, euglobulin clot lysis time, platelet aggregation