fondaparinux and Inflammation

Heparin has been used as an anticoagulant for decades. Recently, attention has been drawn to the non-anticoagulant activities of heparin. Experimentally and clinically those non-anticoagulant properties of heparin inhibit inflammation and metastatic spread of tumor cells. On the molecular level, heparin inhibits the function, expression and/or synthesis of adhesion molecules, cytokines, angiogenic factors and complement. However, despite a similar anticoagulant activity, those non-anticoagulant effects of heparin differ greatly among the different heparin preparations. The same holds true for the most common adverse events of heparin treatment. The incidence of immune mediated heparin-induced thrombocytopenia and cutaneous hypersensitivity responses is greatly, but not exclusively, influenced by the heparin preparation used. As the structure-function relationship of the anti-inflammatory and anti-metastatic effects of heparins are understood in more detail, and as the risk profile of different heparin preparations regarding the induction of adverse events have been identified, we propose to use the different heparin preparations according to the individual needs of each patient.

Topics: Angiogenesis Inhibitors; Anti-Inflammatory Agents, Non-Steroidal; Anticoagulants; Cell Adhesion Molecules; Clinical Trials as Topic; Complement Activation; Cytokines; Fondaparinux; Glucans; Heparin; Humans; Inflammation; Neoplasm Metastasis; Polysaccharides

Surface compatibility with blood is critical both for scientific investigations on hemostasis and clinical applications. Regarding in vitro and ex vivo investigations, minimal alteration in physiological hemostasis is of particular importance to draw reliable conclusions on the human coagulation system. At the same time, artificial coagulation activation must be avoided, which is relevant for the patient, for example to prevent stent graft occlusion. The aim was to evaluate the advantages and disadvantages of antithrombotic and antifouling surface coatings in the context of their suitability for ex vivo incubation and the study of coagulation properties.. We investigated the impact of different protocols for surface coating of synthetic material and different anticoagulants on hemostasis and platelet activation in ex vivo human whole blood. Blood samples from healthy donors were incubated in coated microtubes on a rotating wheel at 37°C. Two protocols for surface coating were analyzed for hemostatic parameters and metabolic status, a heparin-based coating (CHC, Corline Heparin Conjugate) without further anticoagulation and a passivating coating (MPC, 2-methacryloyloxethyl phosphorylcholine) with added anticoagulants (enoxaparin, ENOX; or fondaparinux, FPX). Employing the MPC-based coating, the anticoagulants enoxaparin and fondaparinux were compared regarding their differential effects on plasmatic coagulation by thrombelastometry and on platelet activation by flowcytometry and platelet function assays.. Using the CHC coating, significant coagulation cascade activation was observed, whereas parameters remained mostly unchanged with MPC-based protocols. Extended incubation caused significantly elevated levels of the soluble membrane attack complex. Neither ENOX nor FPX caused a relevant impairment of platelet function or activation capacity and thrombelastometric parameters remained unchanged with both protocols. For translational purposes, we additionally modeled endotoxemia with the MPC-based protocols by incubating with lipopolysaccharide plus/minus thrombin. While coagulation parameters remained unchanged, elevated Interleukin 8 and Matrix Metalloproteinase 9 demonstrated preserved immune cell responsiveness.. The MPC-based protocols demonstrated better hemocompatibility compared to CHC, and ENOX and FPX proved useful for additional anticoagulation. Furthermore, this simple-to-use whole blood model may be useful for experimental analyses of the early coagulatory and immunological response without decalcification.

Topics: Anticoagulants; Enoxaparin; Fondaparinux; Hemostasis; Heparin; Humans; Inflammation

Antithrombin (AT) is a plasma serpin inhibitor that regulates the proteolytic activity of procoagulant proteases of the clotting cascade. In addition to its anticoagulant activity, AT also possesses potent anti-inflammatory properties.. The objective of this study was to investigate the anti-inflammatory activity of wild-type AT (AT-WT) and a reactive centre loop mutant of AT (AT-RCL) which is not capable of inhibiting thrombin.. The cardioprotective activities of AT-WT and AT-RCL were monitored in a mouse model of ischemia/reperfusion (I/R) injury in which the left anterior descending coronary artery was occluded and then released.. We demonstrate that AT markedly reduces myocardial infarct size by a mechanism that is independent of its anticoagulant activity. Thus, AT-RCL attenuated myocardial infarct size to the same extent as AT-WT in this acute injury model. Further studies revealed that AT binds to vascular heparan sulfate proteoglycans via its heparin-binding domain to exert its protective activity as evidenced by the therapeutic AT-binding pentasaccharide (fondaparinux) abrogating the cardioprotective activity of AT and a heparin-site mutant of AT exhibiting no cardioprotective property. We further demonstrate that AT up-regulates the production of prostacyclin in myocardial tissues and inhibits expression of pro-inflammatory cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-6 in vivo by attenuating ischemia/reperfusion-induced JNK and NF-κB signaling pathways.. The present results suggest that both AT and the non-anticoagulant AT-RCL, through their anti-inflammatory signaling effects, elicit potent cardioprotective responses. Thus, AT may have therapeutic potential for treating cardiac I/R injury.

Topics: Animals; Anti-Inflammatory Agents; Antithrombins; Cardiotonic Agents; Fondaparinux; Heparin; Inflammation; Interleukin-6; Ischemia; Leukocytes; Male; MAP Kinase Kinase 4; Mice; Mice, Inbred C57BL; Mutation; Myocardial Ischemia; NF-kappa B; Polysaccharides; Reperfusion Injury; Signal Transduction; Thrombin; Troponin I; Tumor Necrosis Factor-alpha

Topics: Aged; Arginine; Female; Fondaparinux; Heparin; Humans; Immunoenzyme Techniques; Inflammation; Pipecolic Acids; Platelet Factor 4; Polysaccharides; Sulfonamides; Syndrome; Thrombocytopenia; Treatment Outcome

The time course of inflammatory reaction markers in the blood of patients with unstable angina was studied during therapy including arixtra. Plasma concentration of monocytic chemotaxic protein-1 (MCP-1) decreased on days 2 and 3 in patients receiving arixtra and a trend to an increase in MCP-1 concentration was observed on day 7 after the drug was discontinued. After 1 month, MCP-1 level decreased in all patients. The concentration of highly sensitive C-reactive protein also decreased 1 month after the disease onset; no changes in the concentrations of IL-8 and IL-2 receptor α-subunit were detected during these periods. It seems that arixtra is characterized by an anti-inflammatory effect manifesting by reduction of plasma chemokine MCP-1 concentration.

Topics: Aged; Angina, Unstable; Biomarkers; C-Reactive Protein; Chemokine CCL2; Female; Fondaparinux; Humans; Inflammation; Interleukin-2 Receptor alpha Subunit; Interleukin-6; Interleukin-8; Male; Middle Aged; Polysaccharides

Fondaparinux is a synthetic pentasaccharide that selectively inhibits factor Xa (FXa) in an antithrombin-dependent fashion. This newly developed anticoagulant is used in the prevention and treatment of venous thromboembolism. Recently, we showed that fondaparinux reduces inflammation and protects the kidney from ischemia-reperfusion (I/R) injury. However, the relative contributions of the anticoagulant and anti-inflammatory activities of fondaparinux to the observed protection is unknown. To address this, we chemically modified fondaparinux to abolish its affinity for antithrombin and analyzed the effect of this non-anticoagulant (NAC)-pentasaccharide on binding of U937 cells to P-selectin in vitro and on inflammation in a murine model of kidney I/R injury. NAC-pentasaccharide was as effective as fondaparinux at inhibiting the binding of U937 cells to P-selectin. In addition, NAC-pentasaccharide significantly reduced IL-6 and MIP-2 expression and injury in the kidney I/R model. These findings indicate that the anti-inflammatory activity of fondaparinux can be dissociated from its anticoagulant activity and that NAC-pentasaccharide is protective in kidney I/R injury.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Anticoagulants; Blood Coagulation; Cell Adhesion; Chemokine CCL2; Chemokine CXCL2; Chemokines; Creatinine; Disease Models, Animal; Factor Xa Inhibitors; Fondaparinux; Humans; Inflammation; Interleukin-6; Kidney; Kidney Tubules; Macrophages; Mice; Neutrophil Infiltration; Oligosaccharides; P-Selectin; Partial Thromboplastin Time; Polysaccharides; Reperfusion Injury; U937 Cells

Ischemia-reperfusion (I/R) injury is associated with activation of coagulation and inflammation. Interestingly, various anticoagulants have been shown to reduce both coagulation and inflammation in animal models of kidney I/R injury. Fondaparinux is a synthetic pentasaccharide that selectively inhibits factor Xa (FXa) in the coagulation cascade. The aim of this study was to investigate the effect of fondaparinux in a lethal murine model of kidney I/R injury. A murine model of kidney I/R was established. In this model, we measured activation of the coagulation cascade and induction of inflammation. Administration of fondaparinux to I/R-injured mice reduced fibrin deposition in the kidney, reduced serum creatinine levels and increased survival from 0 to 44% compared with saline-treated control mice. Fondaparinux also reduced interleukin-6 and macrophage inflammatory protein-2 expression and decreased neutrophil accumulation in the injured kidneys. Finally, we showed that fondaparinux reduced thioglycollate-induced recruitment of neutrophils into the peritoneum and inhibited the binding of U937 cells to P-selectin in vitro. Our data suggest that fondaparinux reduces kidney I/R injury primarily by inhibiting the recruitment of neutrophils.

Topics: Animals; Blood Coagulation; Cell Movement; Chemokine CXCL2; Creatine; Drug Evaluation, Preclinical; Fibrin; Fondaparinux; Inflammation; Interleukin-6; Kidney; Mice; Models, Animal; Monokines; Neutrophils; Polysaccharides; Reperfusion Injury; Survival Rate