dmp-728 and Thrombosis

Blood platelets play essential roles in normal coagulation and in coronary atherosclerotic disease and its complications. Various antiplatelet therapies, including aspirin, ticlopidine, and dipyridamole, have been developed for use in patients with known coronary artery artery disease to prevent ischemic complications. More recently a more complete anti-aggregation effect has been accomplished by the use of monoclonal antibodies and specific peptide and nonpeptide compounds that bind to the platelet GP IIb/IIIa surface receptor. This receptor becomes activated by platelet stimulation and binds fibrinogen molecules between platelets in the aggregation process. These new antiplatelet drugs are now being evaluated in clinical trials in patients undergoing balloon coronary angioplasty, in whom fewer ischemic events occur when the receptor blocker is used intravenously than with standard therapy, and in patients with stable and unstable angina. Excessive bleeding is an important problem with these agents, and efforts must be made to eliminate this side effect.

Topics: Abciximab; Amino Acid Sequence; Animals; Antibodies, Monoclonal; Arteriosclerosis; Aspirin; Disintegrins; Fibrinolytic Agents; Humans; Immunoglobulin Fab Fragments; Mesylates; Molecular Sequence Data; Myocardial Ischemia; Peptides; Peptides, Cyclic; Platelet Activation; Platelet Adhesiveness; Platelet Aggregation; Platelet Aggregation Inhibitors; Platelet Glycoprotein GPIIb-IIIa Complex; Thrombosis

Either venous or arterial thrombosis is a potentially life-threatening event and existing diagnostic modalities are inadequate to diagnose and to determine the morphology of the evolving thrombus. Thus development of a noninvasive imaging agent that can detect clot location remains a critical and unmet need in nuclear diagnostic medicine. The present study was undertaken to determine the potential of platelet GPIIb/IIIa receptors compared with direct thrombin inhibitors, in the detection of venous and arterial clots.. Initially, the validity of exploiting the degree and extent of specific uptake and retention of a potent GPIIb/IIIa receptor antagonist in venous and in arterial thrombus was confirmed in vitro in artificially created arterial- or venous-type clots, using the radiolabeled antagonist, 3H-DMP728. This was followed by comparing the in-vivo clot/blood distribution of various technetium-99m (99mTc)-labeled, DMP728-derived, GPIIb/IIIa receptor antagonists and of thrombin inhibitors, over time, in mixed arterial/venous or venous clots in arteriovenous shunt and in venous clot models in dogs. In addition, we performed noninvasive single-photon emission tomographic imaging of the venous clot in a deep vein thrombosis model in dogs.. Our data confirmed that potency for the platelet GPIIb/IIIa receptors was maintained after radiolabeling of the parent active GPIIb/IIIa receptor antagonists. DMP728 demonstrated a relatively greater affinity for activated than for unactivated human platelets, which might be essential for attaining an optimal thrombus/blood (target/background) distribution ratio and the optimal detection of small clots (i.e. greater sensitivity).. These data suggest a potential utility of 99mTc-GPIIb/IIIa receptor antagonists, but not of direct thrombin inhibitors, in the diagnosis of venous clots in deep vein thrombosis, pulmonary embolism and arterial thromboembolic disorders including stroke and coronary and peripheral artery thrombotic disorders.

Topics: Analysis of Variance; Animals; Diagnosis, Differential; Disease Models, Animal; Dogs; Humans; Mesylates; Peptides, Cyclic; Platelet Aggregation Inhibitors; Platelet Glycoprotein GPIIb-IIIa Complex; Radionuclide Imaging; Sensitivity and Specificity; Technetium; Thrombophlebitis; Thrombosis

The platelet glycoprotein IIb/IIIa receptor (GPIIb/IIIa, fibrinogen receptor) represents the final common pathway for platelet aggregation. Inhibition of GPIIb/IIIa with antibodies or peptides containing the RGD sequence has been reported to prevent arterial thrombosis. We examined DMP 728 [(cyclic[D-2-amino-butyryl-N2-methyl-L-arginyl-glycyl-L-aspartyl-3-(a min o- methyl-benzoic acid], methanesulfonic acid salt], a cyclic peptidomimetic, GPIIb/IIIa receptor antagonist, for prevention of thrombosis and rethrombosis in a canine model of carotid artery thrombosis. Dogs were anesthetized, and both carotid arteries were instrumented with an electrode, a flow probe, and a stenosis. A 300-microA current was applied to the intimal surface in the right carotid artery (RCA, control) through the electrode; time to occlusive thrombus formation and thrombus mass was noted. The RCA served as the control vessel; the left carotid artery (LCA) served as the test vessel after DMP 728 administration (0.1 or 1. mg/kg, intravenously, i.v.). As compared with controls, occlusive thrombus formation was reduced by both doses of DMP 728 (control 100% n = 12; 0.1 mg/kg i.v. 17%, p < 0.05, n = 6; 1.0 mg/kg i.v. 0%, p < 0.05, n = 6), time to occlusion was increased (p < 0.05), and thrombus weight was reduced (p < 0.05). Ex vivo platelet aggregation was inhibited in all groups. In a second group of animals, a carotid artery thrombus was formed and lysed with anisoylated plasminogen activator complex (APSAC; 0.05 U/kg intraarterially, i.a.) with or without DMP 728.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Carotid Artery Thrombosis; Disease Models, Animal; Dogs; Male; Mesylates; Peptides, Cyclic; Platelet Aggregation Inhibitors; Platelet Membrane Glycoproteins; Recurrence; Thrombosis

This study was undertaken to determine the antithrombotic and thrombolytic efficacy of DMP728 alone and in conjunction with thrombolytic agents.. Coronary artery reocclusion continues to be a significant clinical problem after thrombolytic therapy or balloon angioplasty, with incidence rates of 5-20% regardless of thrombolytic intervention. To date, no adjunctive therapy has been shown to eliminate the incidence of rethrombosis after thrombolysis. DMP728, a novel small-molecular-weight platelet GPIIb-IIIa receptor antagonist, has been shown to prevent rethrombosis after thrombolysis in various arterial thrombosis models in dogs. It might therefore have potential utility in optimizing the clinical outcome of currently available thrombolytic agents. The present investigation was designed to examine the thrombolytic potential of DMP728 alone and in conjunction with different thrombolytic agents.. The deaggregatory effect of DMP728 in reversing human platelet aggregation after initiation of platelet aggregation by 10 mumol/l adenosine 5'-diphosphate was determined using light-transmittance aggregometry. The in-vitro efficacy of DMP728, alone and in combination with thrombolytic drugs, in dispersing a preformed platelet-rich clot was determined using a clot-dispersion assay. In addition, the in-vivo thrombolytic effects of DMP728, alone and in conjunction with streptokinase, were examined in an electrolytically induced femoral artery thrombosis model in dogs.. DMP728 had concentration-dependent deaggregatory and thrombolytic effects in reversing aggregates and in dispersing a preformed platelet-rich thrombus in vitro. Furthermore, it exhibited significant potentiation (P < 0.01) when combined with different thrombolytic drugs such as streptokinase, tissue-type plasminogen activator (t-PA) and urokinase in lysing platelet-rich thrombus. DMP728 had significant in-vivo thrombolytic effects along with synergy in fully restoring arterial flow upon its concomitant use with subeffective to ineffective doses of streptokinase in an electrolytically induced femoral artery thrombosis model in dogs. It also reduced the time to reperfusion and prevented the incidence of rethrombosis after streptokinase.. These findings suggest the potential utility and benefits of DMP728, not only in preventing arterial thrombosis but also in optimizing the efficacy of thrombolytic drugs.

Topics: Adenosine Diphosphate; Animals; Arterial Occlusive Diseases; Clinical Protocols; Dogs; Drug Therapy, Combination; Female; Femoral Artery; Fibrinolysis; Hemodynamics; Humans; Male; Mesylates; Models, Cardiovascular; Peptides, Cyclic; Platelet Aggregation; Platelet Aggregation Inhibitors; Protein Binding; Recurrence; Streptokinase; Thrombosis

1. We studied DMP728, a non-peptide glycoprotein (GP) IIb/IIIa receptor antagonist, for prevention of coronary artery thrombosis or rethrombosis in a chronic canine model subjected to arterial injury. 2. In protocol I, DMP728 (1.0 mg kg-1, i.v., n = 8) or saline (n = 8) was administered and a 150 microA anodal current was applied to the intimal surface of the left circumflex coronary artery (LCX). Dogs were monitored for 6 h and again on each of 5 subsequent days. 3. Ex vivo platelet aggregation was inhibited but returned to baseline 1 day after drug administration. Thrombus weight was reduced (saline, 20.7 +/- 5.0 mg; DMP728 1.7 +/- 0.4 mg; P < 0.05), as was infarct size [saline, 27.5 +/- 4.3; DMP728, 1.6 +/- 0.7 (per cent left ventricle); P < 0.05]. All control animals died by day 3, while all but one of the treated dogs survived the entire protocol (P < 0.05). 4. In protocol II, an LCX thrombus was induced and thrombolytic therapy was initiated 30 min later. DMP728 (1.0 mg kg-1, i.v., n = 8) or saline (n = 8) was administered 5 min after recombinant tissue-type plasminogen activator infusion had begun. The incidence of reocclusion was reduced by DMP728 (saline, 4/8; DMP728, 1/8). One day after thrombolysis, 7/8 DMP728-treated animals were alive compared with 1/8 in the control group (P = 0.01). 5. DMP728 inhibited ex vivo platelet aggregation, prevented primary and secondary occlusive thrombus formation, reduced thrombus weight and infarct size and increased survival in a chronic canine model of coronary artery thrombus formation. DMP728 is an effective anti-platelet intervention when used as the singular adjunctive agent in association with thrombolytic therapy.

Topics: Amino Acid Sequence; Animals; Coronary Vessels; Dogs; Fibrinolytic Agents; Male; Mesylates; Molecular Sequence Data; Myocardial Infarction; Peptides, Cyclic; Platelet Aggregation; Platelet Aggregation Inhibitors; Platelet Membrane Glycoproteins; Recombinant Proteins; Recurrence; Thrombosis; Tissue Plasminogen Activator

Currently used antiplatelet drugs, including aspirin, ticlopidine, and others, are effective against certain but not all of the many endogenous platelet activators. Because of their limited efficacy, a significant number of serious thromboembolic complications still occur, highlighting the need for a more effective therapy. Thus, we have identified a systemically active peptide analogue (DMP 728) of the arginine-glycine-aspartic acid (RGD) recognition sequence that mediates the binding of ligands such as fibrinogen to the platelet glycoprotein (GP) IIb/IIIa receptors. The goals of the present study were to determine the antiplatelet and antithrombotic efficacies of DMP 728 in various arterial thrombosis models.. DMP 728 demonstrated antiplatelet efficacy in vitro in inhibiting ADP-induced human platelet aggregation (IC50, 46 +/- 2 nmol/L) and fibrinogen binding to human platelets (IC50, 2.3 +/- 0.8 nmol/L) or purified human GPIIb/IIIa receptors (IC50, 0.6 +/- 0.1 nmol/L). DMP 728 demonstrated high affinity and specificity for human platelet GPIIb/IIIa over other adhesion molecules. In anesthetized mongrel dogs, DMP 728 at 0.001 to 1.0 mg/kg IV produced dose-dependent antiplatelet effects in inhibiting ex vivo platelet aggregation induced by ADP and in prolonging template bleeding time. DMP 728 effects on bleeding time prolongation were more rapidly reversible than those on platelet aggregation inhibition. A maximal antiplatelet effect for DMP 728 was demonstrated at 0.01 mg/kg IV bolus. The antithrombotic efficacy of DMP 728 was examined in vitro and in vivo after IV administration at different doses in various models of arterial thrombosis. In the coronary artery Folts model in dogs, DMP 728 demonstrated maximal antithrombotic efficacy at 0.01 mg/kg IV bolus with an ED50 of 0.005 mg/kg IV bolus in inhibiting cyclic flow reductions. Additionally, DMP 728 demonstrated 100% prevention of primary thrombosis and rethrombosis (P < .01) after treatment with different thrombolytics, including tissue plasminogen activator and streptokinase, in an electrolytically induced femoral artery thrombosis model in dogs.. Acute intravenous DMP 728 administration (0.001 to 1.0 mg/kg) has dose-dependent antiplatelet and antithrombotic effects in different arterial thrombosis models. These data suggest that DMP 728, a low-molecular-weight GPIIb/IIIa receptor antagonist, may have therapeutic potential as an effective antithrombotic agent in coronary and peripheral artery thromboembolic disorders.

Topics: Angina, Unstable; Animals; Blood Cell Count; Blood Platelets; Coronary Thrombosis; Dogs; Dose-Response Relationship, Drug; Female; Femoral Artery; Fibrinolytic Agents; Humans; In Vitro Techniques; Male; Mesylates; Peptides, Cyclic; Platelet Aggregation Inhibitors; Platelet Membrane Glycoproteins; Rabbits; Thrombosis