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After a 60?min control period of reproducible CFRs (time = ?60?min to 0?min), test agents (saline or Z4A5) were given i.v. as a loading bolus (2?mL) followed by a continuous infusion (60?mL) for 60?min (time = +60?min). Monitoring was continued http://www.selleckchem.com/products/pifithrin-alpha.html for 60?min after the termination of drug infusion (time = +120?min). The antithrombotic effect was quantified by comparing the number of CFRs per hour before, during and after drug infusion. Femoral arterial thrombosis was induced by applying ferric chloride during the infusion from 10?min to 50?min. Four groups were tested in this protocol. Control (physiological saline, n = 6). Low dose Z4A5 (30?��g��kg?1 + 1?��g��kg?1��min?1, n = 6). Middle dose Z4A5 (30?��g��kg?1 + 5?��g��kg?1��min?1, n = 6). High dose Z4A5 (300?��g��kg?1 + 5?��g��kg?1��min?1, n = 6). The hex-peptide Z4A5 was synthesized by solid-phase peptide synthesis methods with standard functionality�Cprotected amino acids. Compounds were purified to 97% purity by HPLC and characterized by NMR, mass spectrometry and amino-acid analysis. The chemical structure of Z4A5 is shown in Figure?2. Z4A5 was dissolved in 0.9% sodium chloride solution for injection. All values are expressed as the mean �� SEM. Data were analysed with the SPSS 11.5 software package for Windows. All variable of distributions were tested for normality with the Kolmogorov�CSmirnov test. Group differences for in vitro platelet aggregation percentage inhibition, thrombus weight and reduction in femoral artery blood flow were performed by one-way anova followed by least-significant difference (LSD) post hoc tests. The IC50s http://www.selleckchem.com/screening/pfizer-licensed-library.html for Z4A5 inhibition of ADP, AA or TH-induced platelet aggregation http://en.wikipedia.org/wiki/Temsirolimus were calculated with Probit Regression. Changes in mean arterial pressure, heart rate, CFR frequency, ex vivo platelet aggregation induced by ADP, coagulation parameters and bleeding time during the experiment were analysed by repeated measure one-way anova followed by LSD post hoc tests. A value of P