초록 ▼

첫째, 고기능성 개비자나무의 우수 집단 선발 및 증식기술 개발에 관한 연구는 1) 천연집단 조사 및 자원파악, 2) 부위별, 개체별, 집단별 함량변이의 정량적인 통계분석 3) 우수집단 선발 및 대량증식기술을 개발하였고,
둘째, 개비자나무를 이용한 신기능성 물질 생산기술 개발에 관한 연구는 1) 기능성 물질의 분석 및 추출, 2) 생리활성물질 분리 및 정제 공정 개발, 3) 공정 가능성 확인 및 제품화 연구를 하였다.
셋째, 개비자나무 유래 생물소재의 in vitro 생리활성 및 독성 분석에 관한 연구는 1) 목본식물 추출

첫째, 고기능성 개비자나무의 우수 집단 선발 및 증식기술 개발에 관한 연구는 1) 천연집단 조사 및 자원파악, 2) 부위별, 개체별, 집단별 함량변이의 정량적인 통계분석 3) 우수집단 선발 및 대량증식기술을 개발하였고,
둘째, 개비자나무를 이용한 신기능성 물질 생산기술 개발에 관한 연구는 1) 기능성 물질의 분석 및 추출, 2) 생리활성물질 분리 및 정제 공정 개발, 3) 공정 가능성 확인 및 제품화 연구를 하였다.
셋째, 개비자나무 유래 생물소재의 in vitro 생리활성 및 독성 분석에 관한 연구는 1) 목본식물 추출물의 항암, in vitro 활성분석, 2) 분리정제된 물질의 in vitro 항암활성 및 독성분석, 3) 항균 활성분석,
4) 특성 분석을 하였으며
넷째, 개비자나무 유래 생물소재의 in vivo 생리활성 및 독성 분석에 관한 연구는 1) in vivo 생리활성 분석 2) in vivo 급성 및 만성 독성 및 조직 검사를 하였다.

Abstract ▼

Plumyew (Cephlotaxus), known for its anti-cancer agents, consists of eight or nine evergreen shrub or tall tree species that are distributed in China and Eastern Asia. Of them Korean native plumyew (Cephlotaxus koreana Nakai) is the only species growing in South Korea. The Korean native plumyew was

Plumyew (Cephlotaxus), known for its anti-cancer agents, consists of eight or nine evergreen shrub or tall tree species that are distributed in China and Eastern Asia. Of them Korean native plumyew (Cephlotaxus koreana Nakai) is the only species growing in South Korea. The Korean native plumyew was studied in eight sites for habitat characteristics and plant samples were analyzed using HPLC (High Performance Liquid Chromatography). The analyses of plant parts showed that the HHT (homoharringtonine) contents were highest in plant needles. HHT contents among and within populations were significantly different with Mt. South Dukyu$(1048{\mu}g/g)$ having the highest and followed by Mt. Naejang$(958.7{\mu}g/g)$ and Mt. Obong$(411{\mu}g/g)$. The relationship between the environmental factors and the HHT contents was investigated that soil moisture appeared tn be the most affecting factor to the content of HHT. The study of mass propagation technique of Korean native plumyew was carried out by means of two experiments : cutting and tissue culture. It was showed that the cutting was feasible to proliferate in short term and also had good effects on root growth. Therefore, cutting propagation was more suitable and practical than tissue culture in terms of time, economics and technique.
A novel purification method was developed to guarantee high purity and yield for producing HHT from Korean native plumyew, The simple and efficient procedure for isolating and purifying HHT from Korean native plumyew biomass consisted of solvent extraction, synthetic adsorbent treatment low-pressure and middle-pressure chromatography, followed by high- pressure chromatography. A purified HHT (purity: 98%) was identified by LC/MS and NMR. The NMR spectrum of HHT was identical to that of HHT reported in the literature. The morphologies of HHT were conveniently controlled by solvent treatment and the monhologies of HHT were characterized by SEM. The residual methanol and methylene chloride were easily removed (to less than concentration limit in ICH(International Conference on Harmonisation) guidance) by spray drying. Amorphous HHT was markedly more soluble in water than crystalline HHT. and the water content of amorphous HHT was less changeable than crystalline HHT during storage. Hence, we found that this procedure is fully possible to make products as medical treatments.
In vitro cytotoxicity assay of the ethanol extract of Korean native plumyew and its active compound, HHT, was carried out using breast and blood cancer cell lines. A breast cancer cell line, SK-BR-3 was more sensitive than SCC-1395. From the cancer cell screening experiment using various blood cancer cell line, K562 cell line was selected because it had the highest concentration-inhibition dependency and sensitivity among the tested cell lines. Using the cell line, the optimal treatment conditions were experimentally obtained. When K562 was treated everyday for 9, 6, 3 days, $IC_{50}$ values of HHT were determined as 0.27, 0.37, and 1.10 mM, respectively. The efficacy of HHT was compared with that of a known anticancer drug compound for blood cancer treatment, adriamycin: the anticancer activity of HHT was comparable to adriamycin. Additionally, the ethanol extract showed no significant adverse effect in hemolysis, and no mutagenicity. The pH stability of HHT in the ethanol extract was examined: HHT was stable in wide range of neutral and acidic pH, but it was unstable in alkaline pH (pH 11). The chemical composition of the ethanol extract was analysed as 1.05% carbohydrate, 0.13% crude lipid, 0.02% crude protein, 0.06% crude fiber, 0.2% crude ash, 98.54% moisture and 5.46% HHT, and the properties of HHT for a medical use, was also determined.
In vivo physiological activity and toxicity test of the ethanol extract from Korean native plumyew was performed. From the acute toxicity test, the number of red blood cell (RBC) in the group treated with HHT was higher than the control group, but there was no significant difference in chronic toxicity test. The degree of damage on liver was serious in acute toxicity test because of the increased hepatic enzymes in chronic toxicity test. In histological analysis of damaged liver, the expansion of hepatocyte and lipid denaturation was observed in acute toxicity test, and abnormal feature of mouse liver surface was also observed in chronic toxicity test HHT did not have effects on the immunological function.

목차 Contents

• Chapter 1. Project summary ... 9
• Chapter 2. Present condition of domestic and foreign country ... 16
• Chapter 3. Contents and results of the project study ... 19
• Section 1. A study on the predominant population selection and mass propagation technique of high-function Korean native plumyew ... 19
• Section 2. A study on development of a novel substance and production method by Korean native plumyew ... 52
• Section 3. A study on the in vitro physiological activity and toxicity test of functional bio-substance derived from Korean native plumyew ... 97
• Section 4. A study on the in vivo physiological activity and toxicity test of functional bio-substance derived from Korean native plumyew ... 126
• Chapter 4. Accomplishment and contribution in research area ... 138
• Chapter 5. Plan to application of study results ... 139
• Chapter 6. References ... 140