Screening And Activity Study Of Anti-virus Leads Against Myocarditis Virus And Ebola Virus

Although the progress of science and technology has changed our living environment and made our life more convenient and comfortable. But the global epidemics of infectious disease constantly remind us that we live in a microbial world, which includes viruses, bacteria and fungi that in an alarming rate to adapt and resist the body’s immune defense mechanism. This overshoot leads to the infectious disease outbreak and epidemic. Therefore, we must accelerate the pace of the development of new drugs, and expect to obtain the drugs with highly effective activity in the prevention and treatment of diseases that caused by pathogens infection. Drug screening is to apply the appropriate screening methods and technologies to extract the pharmacological active compounds from vast amount of compounds. Drug screening is the key to improve the drug research efficiency, accelerate research cycle, cut down research cost and reduce investment risk, which makes the new drug research and development can be sustained. Drug screening new technologies have also become the important content of pharmaceutical research. More and more drug screening technologies have been developed, such as high throughput screening technology, high content screening technology, surface plasm on resonance technology and microfluidic chip technology, etc. The main work of this thesis focuses on the high throughput screening of antiviral drugs in the laboratory.When the viruses contact the host cell surface and release nucleic acid into the cytoplasm, viruses hijack the host of every activity of protein biosynthesis system structure, protein and genome replication, as well as final assembly into progeny viruses released, these processes can be as the target for drug design.Two important study objects in this thesis are Coxsackie virus and Ebola virus, which are RNA viruses and harmful for human health. Myocarditis is a cardiac disease associated with inflammation and injury of the myocardium. Several viruses have been associated with myocarditis in humans. However, Coxsackie virus B3 is still considered the dominant etiological agent. The observed pathology in viral myocarditis is a result of cooperation or teamwork between viral processes and host immune responses at various stages of disease. Both innate and adaptive immune responses are crucial determinants of the severity of myocardial damage, and contribute to the development of chronic myocarditis and dilated cardiomyopathy following acute viral myocarditis. Coxsackie virus B subtypes of 3C protease plays a very important role in the process of Coxsackie virus invasion cell. Ebola virus is a highly virulent pathogen which can cause severe viral hemorrhagic fever in humans and non-human primates, with the case fatality rates being as high as 90%, Because of the high virulence and strong infectivity, currently, there is neither a specific treatment nor a vaccine licensed for use in human. The challenge brought by the Ebola epidemic has promoted the following research focuses:specific and sensitive detection techniques, safe and effective vaccines and drugs. Ebola virus envelope protein GP glycoprotein plays an important role in mediating the process of Ebola virus contacting cell receptors. Therefore, it is important directions of development that against CVB3-3C protease inhibitors and GP inhibitors. In this thesis, we screened 3C protease inhibitors in vitro based on fluorescence resonance energy transfer (FRET) principle and obtained seven good performance of 3C protease with non-competitive and reversible inhibition activity. And Ebola GP inhibitors were screened seven inhibitors of EBOV-GP with specific activity were acquired in vitro.More and more drug screening technologies have been developed. The traditional strategy to deal with multi-drug resistance is to increase the dosage or find another new drug to replace the old one. In this research, we screened antiviral drugs in the laboratory that focused on the high throughput screening to CVB3-3C protease and EBOV-GP. In addition, there was no cytotoxicity for these compounds on HeLa cells in the same dose range. What’s more, the compounds possess novel chemical structures compared to the reported drugs, which suggest that these compounds take effect via a novel mechanism different from the formers. And we can take further study on the compounds based on SAR to achieve better compounds, and even more effective antiviral drugs. This dissertation give us a novel strategy to get inhibitors with new mechanism of action quickly and efficiently, which can provide us novel ideas in drug discovery and next generation antibiotics development.