Optimization of ferric chloride induced thrombosis model in rats: effect of anti-platelet and anti-coagulant drugs.

Numerous studies on various animal species, with variability in the site of application and the concentration of ferric chloride (FeCl3) to induce intravascular thrombosis has prompted us to undertake the present study to obtain a threshold concentration of FeCl3 and validate this model by clinically used anti-thrombotic drugs. A small piece of filter paper, soaked in FeCl3 solution (10, 20, 40, 60 or 80%, w/v), was topically applied on the carotid artery of SD rats to measure the time till occlusion (TTO), to ascertain 20% as an optimal FeCl3 concentration. Anti-platelet drugs, aspirin (30 mg/kg), ticlopidine (200 mg/kg) or clopidogrel (30 mg/kg), administered by oral route for 3 days (once daily) or 4h (single dose) prior to the induction of thrombosis, showed the rank-order: clopidogrel>ticlopidine>aspirin based on TTO by 20% FeCl3. Anti-coagulant drug, heparin (10 U/kg, 30 U/kg and 100 U/kg, i.v) increased TTO in a dose dependent manner (18+/-1.7, 22+/-0.9 and 27+/-3.9 min respectively), while warfarin treated rats at 0.1 mg/kg or 0.3 mg/kg, (po for 5 days), exhibited TTO augmentation to 85+/-11.8 and 120+/-0 min respectively. The results obtained indicate that among the drugs tested warfarin and clopidogrel were most efficacious in this model, while rank order of the other anti-thrombotic drugs were heparin>ticlopidine>aspirin. The present study thus provides a simple, reproducible and well controlled methodology to induce thrombosis in rats by the topical application of 20% FeCl3 to assess the efficacy of new anti-thrombotic agents.

Profile and prevalence of clopidogrel resistance in patients of acute coronary syndrome.

UNLABELLED: Recently reports of a variable platelet response to aspirin and potential resistance to therapy have emerged with thienopyridines group of drugs. However the data available on clopidogrel resistance is scarce. The present study was initiated to prospectively evaluate the prevalence of clopidogrel resistance in patients of acute coronary syndrome(ACS) presently on dual anti platelet therapy by using an established method of optical platelet aggregation. We studied 39 patients of ACS, who were on clopidogrel 300 mg bolus followed by 75 mg per day for 3 days along with aspirin 325 mg per day. Fasting blood samples were assessed using optical platelet aggregation (Chronolog Corp, USA). Clopidogrel resistance was defined as <10% decrease from baseline in platelet aggregation. Clopidogrel semi-responders were defined as 10-29% ( <30%) decrease from baseline in platelet aggregation. Clopidogrel non-responders were defined as a composite of resistant and semi-responders. A baseline mean platelet aggregation obtained from 18 healthy subjects was 63.8 +/- 14.75% with 5 mu and 68.8 +/-13.91% with 10 mu of Adenosine Diphosphate. Hence, the definition of clopidogrel resistance was set as aggregation of >57% with 5 mu ADP and >61.9% with 10 mu ADP (< 10% decrease from baseline). The definition of clopidogrel semi-responder was set as aggregation of >or=45% with 5 mu ADP and >or=48% with 10 mu ADP (10-29% decrease from baseline). The mean platelet aggregation with 5 mu and 10 mu of Adenosine Diphosphate in the patient group was 30.77 +/- 17.19% and 35.71 +/- 17.0% respectively. Based on these criteria, 2.54% patients were found to be clopidogrel resistant, 12.7% were clopidogrel semi-responders and 84.7% were clopidogrel responders. On comparison of clopidogrel responders with non-responders, females ( p=0.07) and patients with higher serum triglyceride levels (p=0.08), had a trend to be more inclined towards clopidogrel non-responders. All other parameters tested namely age, smoking, diabetes, hypertension, obesity, cholesterol, hemoglobin, platelet count, ejection fraction and concurrent drug intake did not show any statistically significant difference among the groups. CONCLUSIONS: This study shows that clopidogrel resistant and clopidogrel semi-responders do occur in Indian patients with ACS and there are no reliable clinical predictors for this condition. The diagnosis therefore relies primarily on laboratory tests.

Baylis-Hillman reaction assisted parallel synthesis of 3,5-disubstituted isoxazoles and their in vivo bioevaluation as antithrombotic agents.

The solution-phase parallel synthesis involving reactions of Baylis-Hillman products of 3-substituted-5-isoxazolecarbaldehydes with nucleophiles and their in vivo antithrombotic evaluations are described along with the results of in vitro platelet aggregation inhibition assay of a few compounds. Results of the detailed evaluation of one of the compounds as an inhibitor of platelet aggregation are also presented.