Porcine epidemic diarrhea virus (PEDV) replication inhibitory coumarins from Saposhnikovia divaricata
- Issued Date
- 2014
- Abstract
- Swine based agro-industries throughout the world are still in big threat of PEDV infection due to lack of efficient prophylactic defenses as well as dependable curing agents. Bioactivity-guided fractionation of methanol soluble extract from radix of Saposnikovia divaricata led to the isolation of three new coumarin glucosides, designated as divaricoumarin A (1), divaricoumarin B (2), divaricoumarin C (3), together with two known coumarins (4-5).Structures of the new isolates (1-3) were established by extensive spectroscopic analysis and absolute configurations were assigned based on ECD calculations. Among all isolates, the known angular pyranocoumarin, hyuganin C (5), revealed strong inhibitory effect on PEDV replication. Quantitative real-time PCR data showed inhibitory effect of 5 on gene responsible for PEDV vital structural protein synthesis (Nucleocapsid, GP2 spike, and GP5 membrane) in a dose-dependent manner. Also, it revealed the inhibitory effect on PEDV nucleocapsid and GP2 spike protein synthesis. The present result indicates the strong potentiality of compound 5 for curing PEDV infection via viral replication inhibition.
- Alternative Title
- 방풍으로부터 분리한 코마린계 화합물의 PEDV 복제 저해활성
- Alternative Author(s)
- Basanta Dhodary
- Department
- 일반대학원 약학과
- Advisor
- Prof. Choi Hong Seok
- Awarded Date
- 2014-08
- Table Of Contents
- Contents ………………………………………………… i
List of Schemes …………………………………………iii
List of Tables ……………………………………………iv
List of Figures …………………………………………v
List of Abbreviations …………………………………vii
Abstract …………………………………………………ix
1. Introduction ………………………………1
1.1. Porcine Epidemic Diarrhea ……………1
1.2. Porcine Epidemic Diarrhea Virus……2
1.3. Diagnostic Tools for PED……………5
1.4. Treatment Measures against PED ……6
1.5. Plant coumarins …………………………6
1.6. Saphosnikovia divaricata…………………8
2. Materials and Methods …………………10
2.1. Materials ……………………………………10
2.1.1. Plant material ………………………………10
2.1.2. Chemicals, reagents and chromatography …10
2.1.3. General experimental procedures ……………11
2.2. Methods ……………………………………………11
2.2.1. Extraction and isolation …………………………11
2.2.2. Acid hydrolysis of compound 1 ………………16
2.2.3. Computational Chemistry for ECD Calculation.16
2.2.4. Cell culture and Virus Stock ……………………16
2.2.5. Cytotoxicity assay…………………………………17
2.2.6. Cytopathic Effect (CPE) inhibition assay ……17
2.2.7. Quantitative Real-time PCR………………………18
2.2.8. Western Blot analysis……………………………18
2.2.9. Immunofluorescence assay……………………19
2.2.10. Statistical analysis………………………………20
3. Results and Discussions ………………………21
3.1. Structural determination of new compounds 1-3 …………………………21
3.1.1. Structural determination of compound 1 ……21
3.1.2. Structural determination of compound 2 ……28
3.1.3. Structural determination of compound 3 ……32
3.2. Anti-PEDV effects of isolated compounds 1-5 ………………37
3.2.1. Assessment of PEDV inhibition via CPE analysis……………37
3.2.2. Assessment of PEDV replication inhibition………………38
4. Conclusions …………………………………44
5. References …………………………………45
6. Acknowledgements ………………………52
- Degree
- Master
- Publisher
- 조선대학교 대학원
- Citation
- 도다리 바산타. (2014). Porcine epidemic diarrhea virus (PEDV) replication inhibitory coumarins from Saposhnikovia divaricata.
- Type
- Dissertation
- URI
- https://oak.chosun.ac.kr/handle/2020.oak/12223
http://chosun.dcollection.net/common/orgView/200000276199
- Appears in Collections:
- General Graduate School > 3. Theses(Master)
- Files in This Item:
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