Coumarins as porcine epidemic diarrhea virus (PEDV) inhibitors from Saposhnikovia divaricata
- Issued Date
- 2013
- Abstract
- PEDV 바이러스에 의한 돼지에서 일어나는 질병은 돼지를 기르는 축산에서 설사를 야기하는 질병을 일으켜, 축산업의 생산성에 심각한 손실을 야기한다. PEDV 바이러스를 저해하는 화합물을 발굴하기 위하여 국내 자생식물 추출물에 대한 탐색을 실시하여, 후보식물로 방풍(Saposhnikovia divaricata, Apiaceae)을 선정하였다.
방풍을 메탄올로 추출한 뒤 농축하여 용매분획을 실시하여 활성을 측정한 결과 PEDV에 대한 저해활성을 보여주는 물질이 EtoAc 층에 있음을 확인하였다. 이후, EtoAc 분획물을 PEDV에 대한 저해활성을 측정하며, 활성물질을 추적하는 방법으로 크로마토 그래피 기법을 사용하여 화합물을 분리하였다. 분리한 화합물 (1-10)의 화학적 구조를 NMR 방법 등을 이용하여 구조 결정하였다. 이후 분리한 화합물 각각에 대하여, 바이러스를 숙주세포에 감염시키고 바이러스에 의한 세포의 사멸사 저해정도를 측정하는 방법인 cytophatic effect (CPE) 실험을 실시하여 각 화합물의 항 바이러스 활성을 측정하였다. 분리된 화합물 중 화합물 2는 화합물은 VERO 세포에 PEDV 유도된 cytopathic 효과에 대해 상당한 저해 활성을 가지고 있는 것으로 밝혔다. 분리된 다른 화합물들인 화합물 1, 3-5번 화합물도 PEDV에 대한 저해활성을 갖고 있지만 높은 농도에서는 세포독성을 갖음을 확인하였다.|The epidemic of PED is grave threat to the swine based agro-economy world-wide and the prophylactic defenses against it are not dependable while there are no agents to treat the infection. This scenario demands for the research to find the solution to this problem. To find the safe, effective and efficient treatment for this virus infection, a plant, Saposhnikovia divaricata (Apiaceae) was chosen, after screening many plant samples. This plant is also a very popular constituent of formulations in Traditional Chinese Medicine. The EtOAc extract from radix of this plant was subjected to bioactivity guided successive chromatographic fractionation, using CPE inhibition based antiviral assay. This led to the isolation of ten coumarin type compounds (1 – 10). Among these, 2 and 6 were found to have significant inhibitory activity (EC50 : 5.91 and 15.26 µM respectively) with good safety index (>3.2) against PEDV induced cytopathic effect in Vero cells. Compound 1, 3, 4 and 5 however showed activity at remarkably small concentration of but they also showed increased degree of cytotoxicity, while remaining coumarins didn’t have significant CPE inhibitory activity
- Alternative Title
- 방풍으로부터 PEDV 바이러스 저해물질로서 코마린 화합물의분리 및 활성 규명
- Alternative Author(s)
- 샤르마 고빈다
- Affiliation
- 조선대학교 대학원
- Department
- 일반대학원 약학과
- Advisor
- 최홍석
- Awarded Date
- 2013-08
- Table Of Contents
- Contents.....................................i
List of schemes........................... iv
List of tables...............................v
List of figures........................................vi
List of abbreviations........................viii
Abstract................................x
1. Introduction............................1
1.1. Porcine epidemic diarrhea (PED)...............................1
1.2. The virus...............................................2
1.2.1. Coronavirus................................3
1.2.2. Evolution......................6
1.2.3. Porcine epidemic diarrhea virus (PEDV).........7
1.3. Vaccines.......................................7
1.4. Saposhnikovia divaricata (Apiaceae)............8
2. Material and Methods..........................12
2.1. Materials.............................................12
2.1.1. Plant material.............12
2.1.2. Chemicals, reagents and chromatography........12
2.2. Methods............................13
2.2.1. Extraction and isolation.......13
2.2.2. Viruses and cell.........16
2.2.3. Cytopathic effect (CPE) inhibition......16
2.2.3.1. Cytopathic effect........16
2.2.3.2. CPE inhibition assay...........17
2.2.3.3. TCID50 (50 % Tissue culture infectious dose).....18
2.2.4. MTT assay.......18
2.2.5. Statistical analysis............19
3. Results and Discussions.....20
3.1. Isolation of compounds from radix of S. divaricata............................20
3.2. Structures of isolated compounds.......20
3.3. Structure determination of isolated compounds............21
3.3.1. Structure determination of compound 1......21
3.3.2. Structure determination of compound 2.....27
3.3.3. Structure determination of compound 3...29
3.3.4. Structure determination of compound 4....31
3.3.5. Structure determination of compound 5...33
3.3.6. Structure determination of compound 6....35
3.3.7. Structure determination of compound 7......36
3.3.8. Structure determination of compound 8...37
3.3.9. Structure determination of compound 9...39
3.3.10. Structure determination of compound 10..40
3.4. Effect of isolated compounds on PED virus induced CPE...............................................42
3.4.1. Determination of TCID50 of stock PED virus suspension....................................42
3.4.2. Assessment of PEDV induced CPE inhibitory activities of isolated compounds…......43
4. Conclusions............48
5. References..........49
6. Acknowledgements..................58
- Degree
- Master
- Publisher
- 조선대학교 대학원
- Citation
- Govinda Sharma. (2013). Coumarins as porcine epidemic diarrhea virus (PEDV) inhibitors from Saposhnikovia divaricata.
- Type
- Dissertation
- URI
- https://oak.chosun.ac.kr/handle/2020.oak/9857
http://chosun.dcollection.net/common/orgView/200000263903
- Appears in Collections:
- General Graduate School > 3. Theses(Master)
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