미성숙 사과의 최종당화산물 생성 저해물질의 동정

국문 초록 (Abstract)

천연물 유래의 당뇨합병증과 관련된 AGEs 생성을 효과적으로 저해하는 활성물질을 개발하기 위하여 본 연구를 수행하였으며, 미성숙 사과 70% EtOH 추출물의 EtOAc 가용부로부터 AGEs 생성 저해활성의 IC50값이 14.7±3.9 μg/mL로 우수한 활성을 확인하였다. 활성을 나타내는 성분의 동정을 위하여 ODS gel 등을 충진제로 활용한 칼럼크로마토그래피를 수행하여 3종의 flavonoid 유도체 및 2종의 caffeoylquinic acid 유도체 화합물을 분리하였고, 각 화합물의 화학구조는 NMR 및 FABMS 스펙트럼 데이터 해석 및 표품과의 HPLC 직접 비교를 통하여 reynoutrin(1), quercitrin(2), isoquercitrin(3), chlorogenic acid methyl ester(4) 및 chlorogenic acid(5)로 동정하였다. 분리된 단일물질의 AGEs 생성 저해활성 평가를 수행한 결과 reynoutrin(1)의 IC50값이 8.6±1.4 μM로 우수한 저해활성을 확인하였다. 또한, quercitrin(2) 및 isoquercitrin(3)의 IC50값이 각각 11.6±2.2 및 81.8±9.6 μM의 활성을 나타냈으며, quercetin 3번 위치에 결합하는 당의 종류에 따라 AGEs 생성 저해활성의 다름이 시사되었다. 2종의 caffeoylquinic acid 유도체화합물인 chlorogenic acid methyl ester(4)와 chlorogenic acid(5)의 IC50값이 각각 75.5±5.3 및 18.6±2.4 μM로 우수한 AGEs 생성 저해활성이 나타났으며, 이들 활성은 methoxy의 결합 여부에 따라 활성이 다름을 확인하였다. 향후 우수한 활성을 나타낸 화합물의 동물실험을 통한 활성기작의 연구가 필요하다고 생각되며, 추가적인 연구를 통하여 기존 유효 성분보다 우수한 AGEs 생성 저해활성을 가지는 새로운 선도화합물 발굴을 위한 기초자료로 활용할 수 있으리라 생각된다.

다국어 초록 (Multilingual Abstract)

Prevention of advanced glycation end products (AGEs) formation is a promising therapeutic strategy for the regulation of diabetic complications. In an effort to discover natural anti-diabetic ingredients from natural food-stuffs, the ethyl acetate (EtOAc)-soluble portion of 70% ethanolic extract from unripe fruits of Malus pumila cv. Fuji was analyzed, and it showed significant inhibitory activity toward AGEs formation. Herein, we present the isolation and identification of three flavonoid derivatives and two caffeoylquinic acid derivatives of a previously known structure, reynoutrin (1), quercitrin (2), isoquercitrin (3), chlorogenic acid methyl ester (4), and chlorogenic acid (5), as well as their inhibitory effects against related diabetic complications using in vitro AGE formation assays. Structures of these compounds were characterized by interpretation of nuclear magnetic resonance and mass spectroscopic data. Among the isolated compounds, reynoutrin (1) and quercitrin (2) exhibited potent inhibitory activities with IC50 values of 8.6±1.4 and 11.6±2.2 μM, respectively. This result suggests that phenolic constituents isolated from M. pumila cv. Fuji might be beneficial for prevention of diabetic complications and related diseases.

목차 (Table of Contents)

• ABSTRACT
• 서론
• 재료 및 방법
• 결과 및 고찰
• 요약
• REFERENCES

참고문헌 (Reference)

1 Yue T, "Ultrasound-assisted extraction, HPLC analysis, and antioxidant activity of polyphenols from unripe apple" 35 : 2138-2145, 2012

2 Fava S, "Role of postprandial hyperglycemia in cardiovascular disease" 6 : 859-872, 2008

3 de Beck PO, "Quercitrin 3'-sulphate from leaves of Leea guinensis" 47 : 1171-1173, 1998

4 Jang DS, "Puerariafuran, a new inhibitor of advanced glycation end products (AGEs) isolated from the roots of Pueraria lobata" 54 : 1315-1317, 2006

5 Kato N, "Polyphenols from flowers of Magnolia coco and their anti-glycation effects" 81 : 1285-1288, 2017

6 Yoshioka Y, "Orally administered apple procyanidins protect against experimental inflammatory bowel disease in mice" 8 : 1802-1807, 2008

7 Edelstein D, "Mechanistic studies of advanced glycosylation end product inhibition by aminoguanidine" 41 : 26-29, 1992

8 Wang CY, "Isolation of fumaric acid, succinic acid, astragalin isoquercitrin and tiliroside from Pteridium aquilinum" 12 : 2298-2299, 1973

9 Nakashima S, "Isolation and structure elucidation of tetrameric procyanidins from unripe apples (Malus pumila cv. Fuji) by NMR spectroscopy" 83 : 144-152, 2012

10 Jung HA, "Inhibitory effects of Nelumbo nucifera leaves on rat lens aldose reductase, advanced glycation endproducts formation, and oxidative stress" 46 : 3818-3826, 2008

11 Vinson JA, "Inhibition of protein glycation and advanced glycation end products by ascorbic acid and other vitamins and nutrients" 7 : 659-663, 1996

12 Lu Y, "Identification and quantification of major polyphenols in apple pomace" 59 : 187-194, 1997

13 Wang F, "Hepatoprotective effect of apple polyphenols against concanavalin A-induced immunological liver injury in mice" 258 : 159-165, 2016

14 Jang DS, "Flavan-3-ols having a gamma-lactam from the roots of Actinidia arguta inhibit the formation of advanced glycation end products in vitro" 57 : 397-400, 2009

15 Yoo NH, "Erigeroflavanone, a flavanone derivative from the flowers of Erigeron annuus with protein glycation and aldose reductase inhibitory activity" 71 : 713-715, 2008

16 박의균, "Effects of Unripe Apple Polyphenols on the Expression of Matrix Metalloproteinase-1 and Type-1 Procollagen in Ultraviolet Irradiated Human Skin Fibroblasts" 한국응용생명화학회 57 (57): 449-455, 2014

17 Lee J, "Chemical constituents from the aerial parts of Aster koraiensis with protein glycation and aldose reductase inhibitory activities" 75 : 267-270, 2012

18 Kim T, "Chemical constituents from Ainsliaea acerifolia as potential anti-obesity agents" 16 : 146-151, 2016

19 Peyroux J, "Advanced glycation endproducts (AGEs): Pharmacological inhibition in diabetes" 54 : 405-419, 2006

20 Son DH, "5-${\alpha}$ Reductase inhibitory effect and astringent activity of green apple rind extract on human keratinocytes and fibroblast cells" 77 : 714-721, 2013

21 Jung YS, "2,3-Dimethoxy-5-methyl-1,4-benzoquinones and 2-methyl-1,4-naphthoquinones: glycation inhibitors with lipid peroxidation activity" 15 : 1125-1129, 2005