Title of Invention

ANTITHROMBOTIC COMPOSITION AND EFFECTIVE METHOD FOR PRESERVING BLOOD EXVIVO

Abstract

ABSTRACT ANTITHROMBOTIC COMPOSITION AND EFFECTIVE METHOD FOR PRESERVING BLOOD EXVIVO The present invention relates to a composition exhibiting enhanced anti-thrombic and thrombolytic effect comprising 5mg/kg of gylcine betaine and a contrasting product selected from ioxaglate (Hexabrix®) and iopamidol. The present invention also relates to an effective method of preserving blood exvivo comprising adding/mixing glycine betaine to the blood to be preserved.

Full Text

This invention relates to the use of glycine betaine to eliminate physiopathological vascular attacks. The invention relates to the curative and preventive activity of glycine betaine in the pathogenesis of thrombo-embolic and haemostatic diseases of arterial or venous origin. Glycine betaine exhibits preventive activity while preventing the formation of thrombi and exhibits a curative activity which prevents the proliferation of thrombi while destroying them. The significance of the present invention is based on the fact that the use of glycine betaine does not result in any risk of haemorrhage or allergy in opposition to the molecules and treatments currently used. PRIOR ART Vascular thromboses are a response of the organism which is facing attack on a vessel wall and on the content of cells and plasma thereof. Thrombosis is a localised activation of coagulation with the formation of a thrombus. The interest to which this pathology has been subjected in recent years has enabled several causative factors to be identified: - The vessel, the vascular wall and the endothelial cells, - The role of elements which occur in blood - The coagulation and fibrinolysis systems and inhibitors thereof. Several types of thromboses exist which can occur in arteries, in veins, in the micro-circulation of the organs, in the cavities of the heart and at artificial surfaces in contact with blood. Vascular thromboses are a response to the attack on the vessel wall and on its content of cells and plasma. A thrombosis is an organised mass of blood elements (platelets, red corpuscles and white corpuscles), of fibrin and of other plasma proteins, which are deposited at the surface or which obstruct the lumen of the vascular system. The mechanisms of thrombosis resemble those of haemostasis, but are pathological due to their abnormal intravascular location. Thromboses and embolisms are the main reasons for clinical complications associated with cardiovascular diseases and atherosclerosis. According to Virchow, at least three types of thrombogenetic factors determine the location, the extent and the regression of a thrombosis: Haemodynamic and rheological factors; Endothelial lesion; Activation of the constituents of blood, particularly of platelets, and of coagulation which results in the formation of thrombin. Thrombo-embolic disease of arterial or venous origin remains one of the main reasons of death in developed countries. Arterial thrombosis is often due to a rupture of the atherosclerotic plaque, whereas venous thrombosis results from a deficit of a coagulation inhibitor (AT III) or from a deficit of a fibrinolysis activator (S protein and/or C protein) or more frequently from stasis. In effect, both of these result from an interaction between blood and the vascular wall, from the formation of a venous thrombosis and/or from a haemostatic anomaly. Arterial thrombosis is more often secondary to a parietal anomaly and mainly involves blood platelets. It contributes to a wide variety of clinical pictures depending on the arterial layers involved in the interruption of vascularisation. Thrombosis is mainly capable of affecting the cardiac arteries (coronary), and the arteries of the lower, cerebral or digestive organs. Thus arterial disease favours the formation of the thrombus itself which is responsible for the majority of terminal vascular occlusions. Moreover, participation of haemostasis disorders and of the thrombus formed at other vascular lesions is evident: aggravation of the lesions of the vascular wall, ischemia and problems in the micro-circulation. Three therapeutic strategies can be distinguished for the prevention of accidents associated with thromboses: Anticoagulants. These constitute the major element in the treatment of a patient exhibiting a thrombo-embolic disorder. Heparin and derivatives thereof are currently used. However, the use of heparins can give rise to two major complications, haemorrhage or thrombopenia. K anti-vitamins (KAV). Prescribed for long-term treatment, these cannot be used in an emergency and cannot be prescribed simultaneously with other antiaggregants, since they potentiate the haemorrhagic effect thereof. Platelet antiaggregants. Prescribed to prevent arterial thrombosis associated with atherosclerosis. The main inhibitors of platelet tunctioning which are currently prescribed are: aspirin, ticlopidine, dipyridamole, and certain non-steroid anti-inflammatory agents such as flurbiprofen and prostacyclin. These treatments are really effective, but have undesirable effects on patients subject to allergies or haemorrhage. Despite their efficacy, all these treatments necessitate special precautions in use, such as the administration of antidotes, overdose problems and unwanted side effects. These treatments make it necessary to monitor patients, due in particular to haemorrhage-related problems which can arise during or after medication, as well as possible incompatibility with other drugs. It was therefore of interest to identify a molecule having a high antithrombotic potential without undesirable effects. Most surprisingly, glycine betaine has been identified as possessing a high therapeutic potential for in the treatment of thromboses. Glycine betaine, or betaine of formula (CH3)3>r - (CH2) - COO", is a molecule known for its osmo-protective properties and for its cosmetic and pharmaceutical uses. Various pharmaceutical uses of betaine are known, particularly the use of betaine for the treatment of homocystinuria, which causes cardiovascular problems (L. & B. Wilken, J. Inher. Metab. Dis. 1997). Thus patients suffering from homocystinuria, which is a genetic anomaly, exhibit premature atherosclerotic and thrombo-embolic disorders (S.H. Mudd et at.. The metabolism and molecular bases of inherited disease, 1995), and of cardiovascular diseases (Mc Cully, Atherosclerosis Rev. 11, 1983). Homocystinuria is a hereditary deficiency, the homozygote form of which is rare. It is estimated that the prevalence of the homozygote form corresponds to 1 in 200 in the general population. Homocystinuria is due to elevated levels of homocysteine in the plasma of the affected patient. The administration of betaine enables the concentration of homocysteine in the blood to be reduced. Publication WO 951 157 50 proposes the use of ingredients comprising betaine in order to prevent vascular disorders in homocystinuric patients. Publication WO 98 /19690 also relates to patients suffering from an elevated homocysteine level in their blood. The use of betaine mnongst other ingredients is intended to reduce the level of homocysteine in the blood, it having been established that homocysteine is a positive factor of risk in the occurrence of cardiovascular diseases, as v^'ell as in Alzheimer's disease. Publication EP 0 347 864 describes the use of betaine together with other ingredients in order to combat the increase in sulfhydryl groups, which are due to cysteine and to homocysteine, in human plasma, and thus to inhibit the formation of atherosclerotic plaques. This anti-atherosclerotic effect is known and is extensively documented. These publications relate to the effect of betaine on the metabolism of lipids (Zapadnyuk et al. Biol. Med. 1987), and on that of cholesterol (Panteleimonovaetal., Farmakol. Toksikol, Moscow 1983). Publication WO 97 38685 describes the use of betaine and taurine for the treatment of complications resulting from ischemia in some organs. Ischemia is a localised stoppage of the bloodstream and only represents one of the pathologies due to thrombosis. Publication EP 0 781 554 comprises examples which describe experiments on enucleated hearts, i.e. on hearts which have been extracted and isolated from the vascular system. The use of betaine for its known osmo-protective and antiradical properties enables the inventors to claim a protective action thereof on the cardiac muscle. Other forms of betaine have been proposed (WO 97 / 06795), but have not hitherto equalled the potency and performance of glycine betaine. None of these publications discloses the potency of glycine betaine with respect to venous and/or arterial thrombosis, nor its anti-aggregant and anticoagulant potency. Glycine betaine, in the context of the present invention can be used for various clinical applications, such as: - Coronary thromboses and venous thromboses - Thromboses and re occlusion of the vascular system following a thrombolysis or an angioplasty - Infarct, angina pectoris, aneurysm, pulmonary embolism, phlebitis - Cerebral embolism - Post-traumatic shock, whether or not of surgical origin - Prevention of accidents of microcirculation in the following cases; haemophilia, chemotherapy, ageing, oral contraception using estrogens, obesity, tobacco addiction, prosthesis, diabetes - Prevention of the risks associated with the administration of contrasting ionic and non ionic products. APPARATUS AND METHOD A/Principle oflaser-induced thrombosis (SeifFge D. et al., 1989; Weichter W. et al., 1983) In this model, lesion of the vascular wall is induced by a laser beam. This beam causes a limited lesion of the vascular endothelium (only I to 2 cells are destroyed). This laying bare of the sub-endothelium, which is a thrombogenetic surface, results in the adherence of platelets via glycoprotein II. This adherence of platelets is followed by the activation thereof They form pseudopods and secrete the content of their granules. This activation results in the appearance of glycoproteins Ilb-IIla which are necessary for the aggregation of the platelets between them. This lesion is induced in the mesenteric microcirculation of the rat. It is immediately followed by the formation of a thrombus (in a few seconds). This thrombus, which rapidly enlarges under the influence of the flow of blood, emboHses before being formed again. By this manes, the assessment of the effect of glycine betaine was conducted pharmacologically in conjunction with the study of two active molecules used as a reference; namely acetylsalicylic acid and heparin (of low molecular weight). The assessment also involved the activity of glycine betaine in relation to the prothrombotic effects induced by contrasting products. B / Stasis-induced thrombosis A laparatomy was performed to operi Uve lower vena cava, on which a ligature was placed at To, followed by subcutaneous injection of glycine betaine at To + 2 hours, followed by withdrawal of the clot and blood samples at To + 6 hours. C/ Experimental procedure Male Wistar rats were used for these tests. They weighed between 200 and 250 grams. After an 8-day stabilisation period, the rats were subjected to fasting for 12 hours. They were than anaesthetised, glycine betaine was administered subcutaneously, and the mesentery (laser) or vena cava (stasis) was opened at the end of the experiments. EXAMPLES: Examplel: Evaluation of the number of emboli and the duration of embolisation after vascular change due to laser firings. Number of embolus or emboli Duration of embolisation (minutes) Negative control NaCl 0.9% 5.33 ±0.58 2±0 Glycine betaine 5 mg/kg 2+0 1±0 Acetylsalicylic acid 100 mg/kg i + l 0.33 ± 0.58 Heparin 2 mg/kg 2.67 + 0.52 1±0 Glycine betaine considerably reduced the number of emboli and the duration of embolisation after vascular change due to laser firings. The results demonstrate its powerful anti-thrombotic activity. Examplel: Evaluation of the bleeding time caused (E. Dejana. Bleeding time in rats. Thrombosis. Rech. 1982) Bleeding time (seconds) Negative control NaCI 0.9% 101.52 ±5.7 Glycine betaine 5 mg/kg 95 ±5 Acelylsalicylic acid 100 mg/kg 276.67 ± 20.82 Heparin 2 mg/kg 313.33 ±20 These results show that glycine betaine maintains the bleeding time which is caused within the values of the negative control. In addition to its anti-thrombotic activity, glycine betaine does not result in any risk of haemorrhage compared with the positive controls. Example 3: Evaluation of platelet aggregation after vascular change due to laser firings (Cardinal & Flower. Pharmacol. Method. 1980) Amplitude (ohms) Velocity (ohms/min) Negative control MaCl 0.9% 13±I 9±I Glycine betaine 5 mg/kg 0.66 ±1.15 1.66±1.15 Acetylsalicylic acid 100 mg/kg 2.33 ± 2.08 2±1 Heparin 2 mg/kg 4.33 ± 0.57 2.66 ± 0.50 These results demonstrate the anti-aggregation effect of glycine betaine. Example 4: Evaluation of the effect on blood cells a / Platelet count Number of platelets (10**) Negative control NaCl 0.9% 788 ±30.14 Glycine betaine 5 mg/kg 804.67 + 20.03 Acetylsalicylic acid 100 mg/kg 855.33 ±63.17 Heparin 2 mg/kg 777.33 ± 6.43 b/ White cell count Number of white cell (lO'^ Negative control NaCl 0.9% 5.03 ± 1 Glycine betaine 5 mg/kg 4.43 ± 0.32 Acetylsalicylic acid 100 mg/kg 4.33 ±1.00 Heparin 2 mg/kg 5.80 ±0.10 c/ Red cell count Number of red cell (10^^) Negative control NaCl 0.9% 6.56 + 0.15 Glycine betaine 5 mg^g 6.19 ±0.25 Acetylsalicylic acid 100 mg/kg 6.15 ±0.31 Heparin 2 mg/kg 6.20 ± 0.20 The counts of the elements occurring in the blood remained within the values of the negative control and demonstrated the innocuousness of glycine betaine Example 5: Biological Balaace a/ Quick time QT (seconds) Negative control NaCl 0.9% 17±1 Glycine betaine 5 mg/kg 16.9 ±1.05 Acetylsalicylic acid 100 mg/kg 18.33 + 2.08 Heparin 2 mg/kg 29.50 + 0.52 b / Activated cephalin time (ACT) ACT (seconds) Negative control NaCI 0.9% 20.5 ±0.5 Glycine belaine 5 mg/kg 39.9 ±1.05 Acetyisalicylic acid 100 mg/kg 27.26+1.1 Heparin 2 mg/kg 39.46 ±1.36 c/ Fibrinogen analysis Fibrinogen (g/!) Negative control NaCl 0.9% 2.45 ±0.19 Glycine betaine 5 mg/kg 1.7 ±0.1 Acetyisalicylic acid 100 mg/kg 2.19 ±0.33 Heparin 2 mg/kg 2.13 ±0.25 d/ Alpba^-antiplasmin analysis (a2AP) a2AP (%) Negative control NaCI 0.9% 30.16 ±0.85 Glycine betaine 5 mg/kg 29.7 ± 0.68 Acetyisalicylic acid 100 mg/kg 29.36 ±0.92 Heparin 2 mg/kg 29.4 ±1.01 e / Antithrombin ni analysis (AT III) AT III (%) Negative control NaCI 0.9% 86 ±3 Glycine betaine 5 mg/kg 89.5 ± 1.37 Acetylsalicylic acid 100 mg/kg 85.33 ±3,51 Heparin 2 mg/kg 77.66 ±1.52 Example 6: Evaluation of tbe activity of glycine l>etaine as a function of time Experimental groups: The product was tested at 5 mg/kg Laser-induced thrombosis Control Group I Group II Group in Group IV NaCl 0.9% The product was injected 1 hour before the experiment The product was injected 2 hours before the experiment The product was injected 3 hours before the experiment The product was injected 4 hours before the experiment a) Effect of the product tested (5 mg/ml/kg) on the bleeding time caused. Group B.T.C. (seconds) NaCi 0.9% 110±21.2 1 105 ±26.2 11 145 ± 15.52 UI 115.5 + 14.2 IV 120 ±10.13 b) Effect of the product tested (5 mg/ml/kg) on arterial thrombosis induced by laser beam Group Number of firings Number of emboli Duration of embolisation (minutes) NaCI 0.9% 2.5 ± 0.84 5.7 ±1.5 2.1 ±0.69 1 3.49+1.07 1.8 ±1.44 0.51 ±0.5 [1 3.0+1.5 1.4±1.I8 0.3 ± 0.23 III 2.5011.25 1.9910.4 1.0010.5 [V 2.7+1.0 2.2 ± 0.69 1.5 ±0.6 c) Effect of the product tested (5 mg/kg) on platelet aggregation induced ex vivo. Group Amplitude (Ohms) Velocity (ohm/minute) NaCl 0.9% 24.23 ± 0.5 14.412.3 [ 11.33 + 3.0S 8.2 + 0.2 II 13.2 ±3.5 9.3 ±1.8 III 12.7±4.1 8.7 ±1.3 IV 13 ±2.8 8.7±1.15 d) Evaluation of the effect of glycine betaine on coagulation factors after repeated administration on 5 days of treatment ACT (seconds) Quick time (seconds) Fibrinogen Untreated control 21.25 ±2.3 16.1 ±1.0 3.03 ± 0.45 Glycine betaine ;5 mg/kg/day) 39.312.3 19.8 ± 1.2 2.2 + 0.1 Example 7: Evaluation of the effect of glycine betaine on venous thrombosis induced by stasis. a) Effect of glycine betaine on clot weight Clot weight (mg) Untreated control 4.033 ±2 Glycine betaine (1 mg/kg) 3.1+0.4 Glycine betaine (2,5 mg/kg) 1.63+0.76 Glycine betaine (5 mg/kg) 0.76 ±0.4 b) Evaluation of the effect of glycine betaine on plasminogen Plasminogen % NaCl 0.9% 2.7 + 0.33 Glycine betaine (5 mg/kg) 1.66 ±0.58 Glycine betaine (2.5 mg/kg) 2 + 0.15 Glycine betaine (1 mg/kg) 2.44 ± 0.58 c) Evaluation of the effect of glycine betaine on coagulation ACT (seconds) Quick time (seconds) Fibrinogen g/1 Untreated control 30.2 ±2.7 16.1 ±1.0 3.03 + 0.45 Glycine betaine (1 mg/kg) 29.1 ±2.3 16.2 ±1.2 2.63 i 0.3 Glycine betaine (2.5 mg/kg) 31.2 ±2.6 16.6 ±0.7 2.2 ±0.17 jlycine betaine (5 mg/kg) 33.5 ± 1-9 15.6 ±0.4 2.32 ± 0.33 d) Evaluation of the effect of glycine betaine on coagulation factors Anti Xa uDits/ml Anti Ua Units/ml Glycine betaine (5 mg/kg) 0.35 ±0.15 - Glycine betaine (2.5 mg/kg) 0.14 + 0.10 - Glycine betaine (1 mg/kg) 0.0810.1 - Treatment with glycine betaine inhibits the thrombo-embolic complications which are initiated by laser firings. In fact, treatment with glycine betaine before laser firings decreases the vascular adherence of platelets and the aggregation thereof. Treatment with glycine betaine inhibits thrombo-embolic complications. In fact, treatment with glycine betaine before the induction of thrombosis exhibited a high antithrombotic potential with regard to all the parameters which come into play in the process of thrombus formation. Moreover, the results for the biological parameters demonstrate the complete innocuousness of glycine betaine, which, in contrast to the reference products used (aspirin and heparin), does not induce any bleeding effect or undesirable side effect. These features mean that glycine betaine in addition to its demonstrated efficacy; can be administered to people at risk of haemorrhage as well as to people who would be subject to risk of sensitivity or allergy if given conventional antithrombotic treatments (haemophiliac, allergic). Glycine betaine does not cause thrombopenia or haemorrhagic disorders (Examples 2 & 4). Experimental result of Example 5c shows that there is a consumption of fibrinogen. It should be noted that, under the same experimental conditions for the preservation of blood, glycine betaine appeared to possess a high anti-coagulant capacity compared with tubes containing heparin or EDTA. The effective dose of glycine betaine appeared to be between 3 and 5 mg per haemolysis tube. This experimental result demonstrates the high anticoagulant potential of glycine betaine. It can thus be claimed that glycine betaine can be used as an anticoagulant both for the treatment of the human body in vivo and for the preservation of blood ex vivo. Evaluation of the activity of glycine betaine compared with that of contrasting products In the context of our research on the anti-thrombotic effects, and in order to complement our preliminary work on the efficacy of glycine betaine, we evaluated the effect of glycine betaine on the increase of the thrombo-embolic risk associated with die use of contrasting products known for their prothrombotic capacities. The significance of this model is that it enables a direct observation to be made of the formation of a thrombus at the site of the vascular lesion. These results explain the occurrence of thrombotic occlusions during angioplasty, especially amongst patients whose endothelium is already damaged or injured. Coronary angioplasty causes a stripping of the endothelium, exposing collagen, elastin and the smooth muscle cells of the circulating blood, analogously to the experimental thrombosis model employed. Thus, there is a higher incidence of new thrombi amongst patients who have had a recent coronary thrombosis or who have an eccentric coronary plaque. The administration of contrasting products reduces the number of white corpuscles, the number of red corpuscles and the number of platelets. Contrasting products interact with leukocytes, induce the liberation of leukotrienes, increase vascular permeability and exert a chemotactic effect. Moreover, contrasting products act to control the expression of P-selectin and cause the adherence of white corpuscles to the vascular endothelium. It has been shown that the use of contrasting products is associated with the occurrence of thrombi in variable amounts depending on the product used. Two contrasting products were studied Hexabrix® (ionic) and ® lopamidol (non-ionic). Example 8; Evaluation of number of emboli and duration of embolisation after vascular changes caused by laser firings and administroo of contrasting products Number of emboli Duration of embolisation (minutes) Negative control NaCl 0.9% 5.33 ±0.58 2±0 Hexabrix* 8±1 3.67 ± 0.58 lopamidol* 11.67 + 0.50 6.33 ±0,52 Glycine betaine 5nig/kg + Hexabrix® 4±1 2 ±0 Glycine betaine 5 mg/ltg + lopamidol* 5.33 ± 0.58 2.33 ± 0.48 Example 8: Evaluation induced bleeding time (IBD) IBD (seconds) Negative control NaCl 0.9% 101.52+5.7 Hexabrix® 195 ±13.23 lopamidoP 128 ±7.64 Glycine betaine 5 mg/kg + Heiuibrix*' 150 ±5 Glycine betaine 5 mg/kg + lopamidol® 111 ± 6.60 Example 10: Evaluation of platelet aggregation after vascular change due to laser firings Amplitude (ohm) Velocity (ohm/min) NaCl 0.9% 13 ±1 9+1 Hexabrix® 6±I 5.66 + 0.57 lopamidol*- 15 ±2.64 12.33 + 0.50 Glycine betaine 5 mg/kg +^ Hexabrix® 2 + 1 5±0 Glycine betaine 5 mg/kg + [opamidol® 4.66 + 0.52 9.33 ± 0.8 Example II: Evaluation of the efiect of glycine betaine on blood cells a / Platelet count Number of platelets (10^ Negative control NaCl 0.9% 788.33 + 30.14 Hexabrix* 620+10 lopamidol* 585.67 ±23.54 Glycine betaine 5 mg/kg + Hexabrix* 669.67 ± 7.37 Glycine betaine 5 mg/kg + lopamidol* 704.33 ±92.33 b/ White cell count Number of white cells (10^ Negative control NaCI 0.9% 5.03 ± 0.20 Hexabrix* 2.96 + 0.21 lopamidol* 3.06 ± 0.35 Glycine betaine 5 mg/kg + Hexabrix* 4.20 ±0.1 Glycine betaine 5 mg/kg + lopamidol* 3.9 ±0.3 c/ Red cell count Number of red cells (10'^) Negative control NaCI 0.9% 6.56 ±0.15 Hexabrix* 5.43 + 0.47 lopamidol* 5.5 ± 0.36 Glycine betaine 5 mg/kg + Hexabrix* 6.5 ±0.15 Glycine betaine 5 mg/kg + lopamidol* 6.6 ±0.19 Example 12: Biological balaace a/ Quick time QT (seconds) Negative control NaCl 0.9% 17±i Hexabrix* 24.13 ±1 [opamidol*' 28.1 ± 0.75 Glycine betaine 5 mg/kg + Hexabrix* 16.36 ± 0.56 Glycine betaine 5 mg/kg lopamidol* 17.83 ±1.2 b/ Activated cephalin time (ACT) ACT (seconds) Negative control NaCl 0.9% 20.5 ± 0.5 Hexabrix* 49.3 ±1.85 lopamidol* 41.33 ± 0.8 Glycine betaine 5 mg/kg + Hexabrix* 25.4 ± 0.61 Glycine betaine 5 mg/kg lopamidol® 22.4 ± 0.7 c/ Fibrinogen analysis Fibrinogen (g/1) Negative control NaCl 0.9% 2.45 ±0.19 Hexabrix* 1.49 ±0.18 lopamidor 1.5 ± 0.8 Glycine betaine 5 mg/kg + Hexabrix* 1.7 + 0.09 Glycine betaine 5 mg/kg lopamidol'^ I.9±0.1 d/ Alpha^-antiplasmin analysis (a2AP) a2AP (%) Negative control NaCl 0.9% 30.16 ±0.85 Hexabrix® 23.26 ±1.06 lopamidol* 25.23 ± 0.95 Glycine betaine 5 mg/kg + Hexabrix** 25.66 ± 0.09 Glycine betaine 5 mg/kg lopamidol* 28.13 ±0.8 e/ Andthrombin III analysis (AT III) ATni(%) Negative control NaCl 0.9% 86.3 ± 3 Hexabrix® 81.63 ±0.66 lopamidol^ 70.6 ±1.51 Glycine betaine 5 mg/kg + Hexabrix^ 79.1 ± 1.05 Glycine betaine 5 mg/kg lopamidol'^ 87.26 ± 0.98 Treatment with glycine betaine inhibits the thrombo-embolic complications associated with the use of contrasting products. In fact, treatment with glycine betaine, before or during the injection of contrasting products, reduces the adherence of platelets and their aggregation at vascular level. These results demonstrate the anti-thrombotic and thrombolytic effects of glycine betaine. It catheters and endothelial lesions due to the administration procedures themselves. Glycine betaine should be notedthe contrasting products can have other side effects such as haemostasis in / catheters and endothelial lesions du remedies these undesirable effects. CONCLUSION Glycine betaine possesses the same, or even better, therapeutic features as those of the anticoagulants and anti-aggregants investigated (acetylsalicylic acid and heparin), whilst exhibiting no undesirable effects. The superior performance as regards therapeutic efficacy of glycine betaine in relation to these two molecules (acetylsalicylic acid and heparin) is an incentive for the formulation of a drug containing glycine betaine as a therapeutically active ingredient, said drug being intended for the treatment of thromboses and tlu-ombo-embolic diseases. According the results presented above, this drug also exhibits anticoagulant, anti-aggregant and fibrinolytic indications. The demonstrated innocuousness of this molecule enables long-term treatments to be considered which do not necessitate biological monitoring. Interest in the use of glycine betaine is based on the fact that it acts at several levels of haemostasis, i.e. it acts on platelet aggregation, coagulation and fibrinolysis. This activity is durable and prevents repeated administration, which constitutes a considerable improvement in relation to existing treatments. The administration of betaine does not induce any haemorrhagic risk or other side effects (e.g. heparin-induced thrombopenia), which constitutes a major advance in antithrombotic therapy. WE CLAIM : 1. A composition exhibiting enhanced anti-thrombic and tlirombolytic effect comprising 5mg/kg of gylcine betaine and a contrasting product selected from oxalate (Hexabrix®) and iopamidol 2. An effective method of preserving blood revive comprising adding/mixing glycine betaine to the blood to be preserved.

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in-pct-2001-1216-che abstract.pdf

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in-pct-2001-1216-che claims.pdf

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in-pct-2001-1216-che petition.pdf