RESUMEN
Objective: To study the influence of different amino group introductions on the antiplatelet aggregation activities of pyridazinone derivatives. Methods: The target compounds were designed and synthesized by inletting different substitution amino groups using acetyl fragment as the connecting segment. Born method was used for preliminary pharmacological test in vitro. Results: Ten target compounds were designed and synthesized; 8 of them were reported firstly and all of them were confirmed by 1HNMR spectra. The results of preliminary pharmacological test showed that all the target compounds exhibited potent antiplatelet aggregation activity. The antiaggregation activities of compounds (6f), (6g), (6h) and (6j) were stong; compounds (6g) and (6j) showed a 5-time higher activity than 6-[4-(pyridin-4-yl-amino) phenyl]-4, 5-dihydro-3 (2H)-pyridazinone (MCI-154). Conclusion: Inletting different substitution amino groups can enhance the antiplatelet aggregation activity of the compounds.
RESUMEN
<p><b>AIM</b>To develop a simple, fast and inexpensive approach as well as an instrument for detection of gene mutation.</p><p><b>METHODS</b>Pyrosequencing based on bioluminometry assay was employed to detect gene mutation. Pyrosequencing is a method of sequencing by synthesis step-by-step using four enzymes, DNA-polymerase, ATP sulfurylase, luciferase and apyrase. The signal was produced by detecting pyrophosphate released during a dNTP incorporation. For mutation detection, a DNA fragment was amplified by PCR at first, followed by a single-stranded DNA preparation. In the second step, a short primer was annealed to the position just before the mutation point. Finally, specific dNTPs were added in terms of the template sequence. The mutation species can be readily determined by the sequence.</p><p><b>RESULTS</b>A new instrument was developed for gene mutation detection by pyrosequencing. To iteratively inject small amount of each dNTP for the sequencing reaction, capillaries were used to connect dNTP reservoirs and the reaction chamber. Each dNTP was delivered by adding a gas pressure on the top of a corresponding dNTP reservoir, by which 0.2 microL of dNTP can be exactly added each time. It was theoretically proved that undesired liquid seep through the capillary did not affect the sequencing reactions in pyrosequencing. In addition, the three possible variants (wildtype, mutant and heterozygote) of a mutant point Cys275Ser in P53 gene exon 8 were determined by pyrosequencing using the instrument. A simple method was also described for rapidly distinguishing the type of a variant.</p><p><b>CONCLUSION</b>The developed method is very simple, and the corresponding instrument is inexpensive and easy to operate, which can be used to detect many types of mutation.</p>