Anti-influenza Activity Study Of Neuraminidase Inhibitors

The influenza virus belongs to the Orthomyxoviridae family.It is divided into two categories of human and animal.And the human influenza has three subtypes(A,B and C).Influenza A virus is the most frequent and harmful in this three subtypes.It had broken several times of influenza in the worldwide.This research aimed at establishing an efficient and stable detection system to test the neuraminidase inhibitory activity of anti-flu drugs.Then we used this detection system to test the neuraminidase inhibitory activity of many sialic acid analogues and cyclohexene analogues.This study is divided into three parts,the first part is the preparation of MUNANA.Using MUNANA to detect neuraminidase inhibitory activity is the most common method.But this substrate is very expensive,if we use it to do massive screening,the cost will be too high.Thus,we designed route to use cheap raw materials,through carboxyl methyl esterification,hydroxyl acetylation,terminal of chloride,phenolic glycosides,deacetylation and de methyl esterification reaction,to produce MUNANA,make massive screening possible.The second part is the preparation of influenza virus.Like the substrate,the massive screening will also cost much neuraminidase,which provide from influenza virus.So,to establish a stable and productive system to prepare influenza virus is necessary.After several experiments,we decided to use SPF embryonated eggs to prepare influenza virus.And we successfully prepared three strains of influenza virus(A/Hunan/SWL1331/2014 H1NlM A/Chiken/Beijing/AT609/2014 H9N2 and A/Puerto Rico/8/1934 HlNl).The last part is to use the substrate and influenza virus that we prepare to test the neuraminidase inhibitory activity.After a large number of experiments,we optimized the conditions(incubation temperature,pH,Ca2+ concentration,etc.)of this detection system.And we tested the neuraminidase inhibitory activities of many sialic acid analogues and cyclohexene analogues by using this improved detection system.In summary,a stable and rapid detection system for testing the neuraminidase inhibitory activity was established,which could be applied for massive drug screening.