Design and Synthesis of Human Norovirus Inhibitors Containing Stilbenoids = 항노로바이러스 제제로서 Stilbene 유도체의 발견 및 개발

다국어 초록 (Multilingual Abstract)

The Norovirus (NV) is a major cause of viral gastrointestinal infections. Human norovirus, belongs to the genus norovirus, is highly contagious. So far, there are no specific anti-NV drugs or vaccines to treat NV infections. Progress in the region has been largely hampered because the immune response to human NV is not well understood. Therefore, it is urgent to design and develop HNV treatments. The first total synthesis of gramistilbenoids A-C has been reported in our lab. We screened the chemicals for anti-norovirus activity using qPCR assay to HG23 cell line. The structure-activity relationship of gramistilbenoids was investigated. A series of stilbene analogs were synthesized starting from vinyl stilbene compound. Among these newly synthesized compounds, several amide derivatives of vinyl stilbenes exhibited potent anti-NV activity with EC50 values ranging from 1 to 2 μM. A trans-vinyl stilbenoid with an appended substituted piperazine amide, exhibited potent anti-NV activity and also displayed favorable metabolic stability.

목차 (Table of Contents)

• 1. INTRODUCTION 1
• 1.1 Genome and Structure of Norovirus 4
• 1.2 The Norovirus Life Cycle 7
• 1.3 Model Systems for the Study of Norovirus Gene Expression and Replication 10
• 1.3.1 Surrogate Viruses 10
• 1.3.2 In Vitro Cell Cultured Model 10
• 1.3.3 Animal Model 11
• 1.4 Strategies and Targets for Anti-Norovirus Drug Discovery 14
• 1.4.1 Vaccines 14
• 1.4.2 Targeting Viral Entry (Glycan Binding Site as a Potential Target) 15
• 1.4.3 Non-Structural Proteins as Targets 16
• a) HNV Protease 17
• b) HNV RdRp 19
• 1.4.4 Interferons and Targeting Host Factors as Antiviral Approach 24
• 1.4.5 Miscellaneous Compounds 25
• 1.4.6 Anti-Norovirus Drugs in the Clinical Development Stage 26
• 2. RESULTS AND DISCUSSIONS 29
• 2.1 Synthesis 29
• 2.1.1 Modification of Hydroxyethyl Moiety of Compound 8 29
• 2.1.2 Synthesis of Stilbenes 66-72 30
• 2.1.3 Synthesis of Vinyl Stilbenes 76a-e and Its Ester Analogues 77a-c 31
• 2.1.4 Synthesis of Amide Analogues 78a-m 32
• 2.1.5 Synthesis of cis-Analogue 83 33
• 2.1.6 Synthesis of 1,2-Diarylethane Analogue 87 36
• 2.2 Biological Evaluation 37
• 2.2.1 Structure-Activity Relationship (SAR) Study 37
• 2.2.2 Calculated Physiochemical Properties and Preliminary ADME Data of Compounds 76b, 78e, 78f and 78k 49
• 2.2.3 PAMPA and Liver Microsomal Stability Assay 50
• 3. EXPERIMENTAL SECTION 52
• 3.1 Chemistry 52
• 3.1.1 General Experimental Procedures 52
• 3.1.2 General Procedure for Arbuzov Reaction (65 and 74a-d) 56
• 3.1.3 General Procedure for HWE Olefination (66-68) 56
• 3.1.4 General Procedure for Stille Coupling (69-71) 59
• 3.1.5 Synthesis of Compounds 75a-e (HWE Olefination) 63
• 3.1.6 Synthesis of Compounds 76a-d (Stille Coupling) 66
• 3.1.7 General Procedure for O-alkylation (77a-c) 70
• 3.1.8 General Procedure for Amide Coupling (78a-m) 73
• 3.2 Biology 88
• 3.2.1 Measurement of Cytotoxicity 88
• 3.2.2 Quantitative Real-Time-RT-PCR (qRT-PCR) Analysis 88
• 3.2.3 Western blot analysis 89
• 3.2.4 HNV RdRp assay 89
• 3.2.5 Microsomal Stability 90
• 4. CONCLUSION 92
• REFERENCES 94