ABSTRACT
The possibility of NQ12 (2-chloro-3-[4-(ethylcarboxy)-phenyl]-amino-1,4-naphthoquinone) as a novel antithrombotic agent and its mode of action were investigated. The effects of NQ12 on platelet aggregation in human platelet-rich plasma in vitro, in rats ex vivo, and on murine pulmonary thrombosis in vivo, as well as the mode of antithrombotic action were examined. NQ12 potently inhibited ADP-, collagen-, epinephrine-, and calcium ionophore-induced human platelet aggregations in vitro concentration-dependently. NQ12 significantly inhibited rat platelet aggregation in an ex vivo study. NQ12 prevented murine pulmonary thrombosis in a dose-dependent manner. However, NQ12 did not affect coagulation parameters such as activated partial thromboplastin time, prothrombin time, and thrombin time. NQ12 inhibited fibrinogen binding to the platelet surface GPIIb/IIIa receptor, but failed to inhibit binding to the purified GPIIb/IIIa receptor. Thromboxane B(2) formation caused by thrombin or collagen was inhibited significantly by NQ12. The phosphoinositide breakdown induced by thrombin or collagen was inhibited concentration-dependently by NQ12. These results suggest that NQ12 may be a promising antithrombotic agent, and its antithrombotic activity may be due to antiplatelet aggregation activity, which may result from the inhibition of phosphoinositide breakdown and thromboxane A(2) formation.
Subject(s)
Fibrinolytic Agents/pharmacology , Naphthalenes/pharmacology , Platelet Aggregation Inhibitors/pharmacology , Platelet Aggregation/drug effects , Animals , Blood Coagulation/drug effects , Blood Platelets/drug effects , Blood Platelets/metabolism , Disease Models, Animal , Fibrinolytic Agents/chemical synthesis , Fibrinolytic Agents/chemistry , Fibrinolytic Agents/therapeutic use , Inositol Phosphates/metabolism , Male , Mice , Mice, Inbred ICR , Naphthalenes/chemical synthesis , Naphthalenes/chemistry , Naphthalenes/therapeutic use , Platelet Aggregation Inhibitors/chemical synthesis , Platelet Aggregation Inhibitors/chemistry , Platelet Aggregation Inhibitors/therapeutic use , Platelet Glycoprotein GPIIb-IIIa Complex/drug effects , Platelet Glycoprotein GPIIb-IIIa Complex/metabolism , Pulmonary Embolism/prevention & control , Rats , Rats, Sprague-Dawley , Thromboxane B2/metabolismABSTRACT
The effect of p6304 (2-chloro-3-(4-hexylphenyl)-amino-1,4-naphthoquinone) as a novel antithrombotic agent was investigated. NQ304 was found to inhibit platelet aggregation in human platelets in vitro and in rat ex vivo, and murine pulmonary thrombosis in vivo. NQ304 potently inhibited adenosine diphosphate (ADP), collagen, epinephrine and calcium ionophore-induced human platelet aggregation in vitro dose-dependently. In the ex vivo study, oral administration of NQ304 significantly inhibited platelet aggregation in rats. However, NQ304 was found not to affect the coagulation system, since it did not change the prothrombin time (PT), activated partial thromboplastin time (APTT) and thrombin time (TT). The agent prevented death due to pulmonary thrombosis by the platelet aggregates in mice in vivo. In the mouse tail bleeding time test, NQ304 showed a significant prolongation of the tail bleeding time in conscious mice. These results suggest that a principal antithrombotic effect of NQ304 may be due to the antiplatelet aggregation activity but not to anticoagulation activity.
Subject(s)
Fibrinolytic Agents/pharmacology , Naphthoquinones/pharmacology , Platelet Aggregation Inhibitors/pharmacology , Animals , Bleeding Time , Blood Coagulation/drug effects , Blood Platelets/drug effects , Cell Survival/drug effects , Humans , In Vitro Techniques , L-Lactate Dehydrogenase/blood , Male , Mice , Mice, Inbred ICR , Platelet Aggregation/drug effects , Rats , Rats, Sprague-DawleyABSTRACT
The antithrombotic activities and mode of action of green tea catechins (GTC) and (-)-epigallocatechin gallate (EGCG), a major compound of GTC, were investigated. Effects of GTC and EGCG on the murine pulmonary thrombosis in vivo, human platelet aggregation in vitro, and ex vivo, and coagulation parameters were examined. GTC and EGCG prevented death caused by pulmonary thrombosis in mice in vivo in a dose-dependent manner. They significantly prolonged the mouse tail bleeding time of conscious mice. They inhibited adenosine diphosphate- and collagen-induced rat platelet aggregation ex vivo in a dose-dependent manner. GTC and EGCG inhibited ADP-, collagen-, epinephrine-, and calcium ionophore A23187-induced human platelet aggregation in vitro dose dependently. However, they did not change the coagulation parameters such as activated partial thromboplastin time, prothrombin time, and thrombin time using human citrated plasma. These results suggest that GTC and EGCG have the antithrombotic activities and the modes of antithrombotic action may be due to the antiplatelet activities, but not to anticoagulation activities.
Subject(s)
Catechin/analogs & derivatives , Catechin/pharmacology , Fibrinolytic Agents/pharmacology , Platelet Aggregation Inhibitors/pharmacology , Platelet Aggregation/drug effects , Tea/chemistry , Adenosine Diphosphate/antagonists & inhibitors , Adenosine Diphosphate/pharmacology , Animals , Aspirin/pharmacology , Bleeding Time , Calcimycin/pharmacology , Calcium/blood , Collagen/antagonists & inhibitors , Collagen/pharmacology , Epinephrine/antagonists & inhibitors , Epinephrine/pharmacology , Humans , Ion Transport/drug effects , Ionophores/pharmacology , Male , Mice , Mice, Inbred ICR , Platelet Aggregation Inhibitors/isolation & purification , Pulmonary Embolism/drug therapy , Rats , Rats, Sprague-DawleyABSTRACT
The antiplatelet and antithrombotic activities of a newly synthesized NQ301, 2-chloro-3-(4-acetophenyl)-amino-1,4-naphthoquinone, were investigated on human platelet aggregation in vitro and rats ex vivo, and murine pulmonary thrombosis in vivo. NQ301 potently inhibited ADP-, collagen-, epinephrine- and calcium ionophore A23187-induced human platelet aggregation in a concentration-dependent manner in vitro. NQ301 significantly inhibited platelet aggregation in orally administered rats ex vivo. NQ301 prevented death due to pulmonary thrombosis in mice dose-dependently in vivo. NQ301 also showed significant prolongation of tail bleeding time in conscious mice. However, NQ301 did not alter such coagulation parameters as activated partial thromboplastin time, prothrombin time, and thrombin time in human plasma. These results suggest that NQ301 may be a promising antithrombotic agent, and the antithrombotic activity of NQ301 may be due to antiplatelet aggregation activity but not to in vitro anticoagulation.
