초록 ▼

노인성 당뇨병의 약물 목표로 Sirtuin, PTP1B 및 AMPK를 활성화하는 물질의 탐색이 시도 되었으며 주관기관에서 발굴된 DDAH (dimethylarginine dimethylaminohydrolase), ANT-1 (adenine nucleotide translocase)의 promotor를 활성화하는 물질을 발굴하기 위하여 국내 및 국외의 식물추출물로부터 발굴이 시도 되었다.

3단계에서 오랜 기간 약재로서 사용되어 온 인삼으로부터 SIRTs를 활성화시키는 화합물의 발굴이 시도 되었다. SIRTs의 활성을 측

노인성 당뇨병의 약물 목표로 Sirtuin, PTP1B 및 AMPK를 활성화하는 물질의 탐색이 시도 되었으며 주관기관에서 발굴된 DDAH (dimethylarginine dimethylaminohydrolase), ANT-1 (adenine nucleotide translocase)의 promotor를 활성화하는 물질을 발굴하기 위하여 국내 및 국외의 식물추출물로부터 발굴이 시도 되었다.

3단계에서 오랜 기간 약재로서 사용되어 온 인삼으로부터 SIRTs를 활성화시키는 화합물의 발굴이 시도 되었다. SIRTs의 활성을 측정하기 위하여 SIRT1과 p53을 co-transfection 시켜서 이들의 활성을 측정하는 방법을 확립하고 추가적으로 in vitro SIRT1 deacetylation 측정법, NAD+/NADH 비율을 측정하는 방법 등을 확립하였다. 이러한 방법 등을 사용하여 인삼으로부터 3개의 신규화합물을 포함하여 5개의 화합물을 SIRT 활성화물질로 규명하였다. 신규 화합물인 파녹솔 A (Panaxol A, 화합물 1), 파녹솔 B (Panaxol B, 화합물 2), 파녹솔 C (Panaxol C, 화합물 3)과 기지물질인 Dammar-20(22),24-diene-3β,6α,12β-triol (화합물 4) 및 20(S)-Ginsenoside Rg₃ (5)로 결정되었다.

E. abyssinica으로부터 13종 화합물을 분리하고 그들의 구조를 분석하였다. 13종의 분리한 화합물 중 신규 화합물 1, 6, 9, 12 및 13의 화합물의 구조는 추가적인 핵자기공명 분석을 통하여 결정하였다.
분리된 13종의 화합물에 대한 PTP1B 에 대한 저해 활성을 측정한 결과, 화합물 6, 10 과 12 화합물을 제외하고는 14.9 ± 1.6에서 98.1 ± 11.3 μM에서 저해작용을 나타내었다. AMPK 활성화 물질로서 베트남 식물인 Morinda longissima로부터 2단계에서 특이적으로 9,10-anthraquinone의 존재가 확인되어 보고한 ML.365 물질 8, 18, 19, Modasama A 및 B외에 추가적으로 2-3차년도에 15종의 화합물을 분리정제하고 구조를 확인하였다. 화합물들의 10 μM 농도에서 p-AMPK 및 p-ACC를 활성화 작용이 관찰되었으며,상대적으로 화합물 2번의 AMPK 활성화 작용이 강하였다.

( 출처 : 요약서 3p )

Abstract ▼

Diabetes mellitus is one of the most prevalent and serious metabolic diseases and the principal cause of morbidity and mortality in the human. In diabetes, there is a failure to increase glucose uptake into peripheral tissues in response to insulin, leading to chronically elevated levels of glucose

Diabetes mellitus is one of the most prevalent and serious metabolic diseases and the principal cause of morbidity and mortality in the human. In diabetes, there is a failure to increase glucose uptake into peripheral tissues in response to insulin, leading to chronically elevated levels of glucose in the circulation. In type 2 diabetes, hyperglycemia is one of the characteristic pathogenesis, and it is clear that control of hyperglycemia can attenuate the development of chronic complications such as retinopathy and nephropathy. Therefore, therapy for type 2 diabetes depends on mainly several approaches to reduce the hyperglycemia itself. As drug targets of elderly diabetes, we has been studied on SIRT1 and AMPK activators,PTP1B inhibitors, and also DDAH and ANT-1 which were suggested by project coordinator as chronic complications in many elderly peoples.

SIRT1, a homologue of silent information regulator two (SIR2), SIRT1 plays a pivotal role in a variety of cellular processes including cellular ageing, neuronal protection, and anti-apoptosis by deacetylating non-histone proteins, such as the forkhead transcription factor(FOXO), the RelA/p65 subunit of nuclear factor-kappa B (NF-kB), and tumor suppressor p53.^4,5Once activated, SIRT1 can promote the potentiation of stress resistance and extend longevity through age-related metabolic regulation, including regulation of factors involved in cell survival, cardioprotection, neuroprotection, insulin secretion, glycolysis, and fat storage.Therefore, finding effective SIRT1 activators may be significant for combating the aging process, and for treating age-related diseases. During a search for SIRT1 activators, three new dammarane triterpenes, 6α,20(S)-dihydroxydammar-3,12-dione-24-ene (1), 6α,20(S),24(S)-trihydroxydammar-3,12-dione-25-ene (2), and 6α,20(S),25-trihydroxydammar-3,12-dione-23-ene (3), as well as two known triterpenes, dammar-20(22),24-diene-3β,6α,12β-triol (4) and 20(S)-ginsenoside Rg3(5), were isolated from P anax ginseng leaves. Compounds 1 and 3-5 showed potential as SIRT1 activators, as analyzed by in vitro enzyme-based SIRT1-NAD/NADH and SIRT1-p53 luciferase cell-based assays. They were also found to increase the level of NAD+/NADH ratio in HEK293 cells.

PTP1B plays a critical role in regulating glucose homeostasis and body weight by act ing as a key negative regulator of insulin and leptin signaling pathway, respectively. Its overexpression has been shown to inhibit the insulin receptor (IR) signaling cascade, and increased expression of PTP1B occurs in insulin-resistant states, while PTP1B knockout mice have been shown to increase insulin sensitivity and obesity resistance. Furthermore, recent evidence has shown that the leptin signaling pathway can be attenuated by PTPs, and there are many reports that PTP1B is also involved in this process. In the course of our program to search for protein tyrosine phosphatase 1B (PTPB) inhibitors, five new 5-deoxyflavonoids along with eight known derivatives were isolated from EtOAc layer of the root bark of Erythrina abyssinica. Their structures were elucidated on the basis of spectroscopic (IR, UV, MS, CD, 1D− and 2D−NMR) and physicochemical analyses. All isolates exhibited moderate inhibitory effects on the enzyme assay with IC₅₀ values ranging from 14.9 ± 1.6 to 98.1 ± 11.3 μM. Compounds with prenyl and methoxy groups in the B ring (1, 2, 4, 8, and 13) possessed strong activity (IC₅₀ 14.9 ± 1.6 to 19.2 ± 1.1 μM), while compounds (3, 5, and 9) with 2,2-dimethylpyrano ring showed less inhibitory effect (IC₅₀ 22.6 ± 2.3 to 72.9 ± 9.7 μM).

The AMP-activated protein kinase (AMPK), a heterotrimeric functional enzyme complex, plays a key role in metabolic control and regulating cellular energy homeostasis. The AMPK activation on the phosphorylation of Thr¹⁷² at the catalytic α subunit can be regulated by calmodulin-dependent protein kinase kinases (CaMKKs) and serine/threonine kinase(LKB1), which are two upstream kinases, and also can be positively influenced by the intracellular AMP/ATP ratio. The activated AMPK is able to regulate lipid homeostasis, and this function has been regarded as a promising strategy for the treatment of type 2 diabetes due to the strong association between the deregulation of the fatty acid metabolism and the development of insulin resistance. Bioassay-guided fractionation of an ethanol extract of the roots of Morinda longissima, using 2-NBDG assay in 3T3-L1 adipocyte cells, yielded two new anthraquinones, modasimas A (10) and B (20), along with eighteen known analogues (1-9 and 11-19). These anthraquinones showed the potential action as insulin mimetic to improve glucose uptake via activation of AMPK.

( 출처 : SUMMARY 15p )

목차 Contents

• 표지 ... 1제 출 문 ... 2보고서 요약서 ... 3요 약 문 ... 4SUMMARY ... 15CONTENTS ... 17목차 ... 18제1장. 연구개발 과제의 개요 ... 19제2장. 국내·외 기술개발 현황 ... 31제3장. 연구개발수행 내용 및 결과 ... 34 3.1. P anax ginseng 의 잎으로부터 신규 SIRT1 활성물질의 분리 ... 34 3.2. P anax ginseng 의 잎으로부터 신규 SIRT1 활성물질의 구조 결정 ... 35 3.3. P anax ginseng 의 잎으로부터 SIRT1 활성물질의 SIRT1 활성측정 ... 50 3.5. E rythrina abyssinica 의 뿌리로부터 신규 PTP1B 활성물질의 분리 ... 54 3.5. E rythrina abyssinica 의 뿌리로부터 신규 PTP1B 활성물질의 구조 결정 ... 55 3.6. E rythrina abyssinica 로부터 화합물의 PTP1B 저해 활성 ... 74 3.7. M orinda longissima 로부터 9,10-anthraquinones 화합물의 분리 ... 76 3.8. M orinda longissima 로부터 9,10-anthraquinones 화합물의 구조 결정 ... 77 3-9. M orinda longissima 로부터 9,10-anthraquinones 화합물의 AMPK 활성 측정 ... 81제4장. 목표달성도 및 관련분야에의 기여도 ... 83제5장. 연구개발 결과의 활용계획 ... 88제6장. 연구개발과정에서 수집한 해외과학기술정보 ... 90제7장. 연구시설 및 장비현황 ... 92제8장. 참고문헌 ... 93끝페이지 ... 96