초록 ▼

해양생물로부터 분리되어 구조 규명된 화합물, 그 합성유도체 및 해양생물 추출물 등 800여종의 시료에 대한 간암, 위암, 대장암, 폐암, 유방암, 혈액암 세포주, pacritaxel 내성 폐암세포주에 대해 성장 억제활성을 평가하여 고역가 hit 화합물 61종을 도출하였음. 특히, halichondramide 유도체, wondonin 유도체, psammaplin A 유도체, F452-R (Asperphenin), Salternamide A 등의 Hit compounds를 선정하여 세포내 신호전달체계 조절 평가 및 동물실험을 진행하여

해양생물로부터 분리되어 구조 규명된 화합물, 그 합성유도체 및 해양생물 추출물 등 800여종의 시료에 대한 간암, 위암, 대장암, 폐암, 유방암, 혈액암 세포주, pacritaxel 내성 폐암세포주에 대해 성장 억제활성을 평가하여 고역가 hit 화합물 61종을 도출하였음. 특히, halichondramide 유도체, wondonin 유도체, psammaplin A 유도체, F452-R (Asperphenin), Salternamide A 등의 Hit compounds를 선정하여 세포내 신호전달체계 조절 평가 및 동물실험을 진행하여 항암 효능을 평가하였음. 또한, 확보된 1,200여종의 시료에 대한 세균, 항생제 내성균 MRSA, 진균, 항균표적효소 (SrtA, ICL, Na⁺ / K⁺ ATPase, Actin) 저해 활성을 평가하여 고역가 hit 화합물 65종을 도출하였음. 특히 psammaplin A 합성유도체 4종은 세균 및 MRSA에 대해 vancomycin 등 기존의 항생물질과 동정도의 강한 저해활성을 나타내었음. 구조-활성 평가를 완료한 결과 및 물질 다량확보 결과를 토대로 2 종류의 물질에 대해 in vivo 감염동물모델에서 활성을 확인한 결과 10 mg/kg 농도에서 매우 우수한 생존율을 보였음. 또한, 우수한 활성물질 및 그의 유도체에 대해서는 HPLC 분석법을 확립하고 validation한 후, 기본적인 물성평가와 더불어 in vivo pharmacokinetic study를 진행하였음.

Abstract ▼

Ⅳ. Results of the Study
1. 339 kinds of Marine natural products and synthetic derivitavies and 507 kinds of marine organism extracts, 846 kinds in total, were screene using SRB and MTT assay on stomach, lung, colon, blood, breast, liver, and pacritaxel-resistant lung(A549-Pa) cancer cells to eval

Ⅳ. Results of the Study
1. 339 kinds of Marine natural products and synthetic derivitavies and 507 kinds of marine organism extracts, 846 kinds in total, were screene using SRB and MTT assay on stomach, lung, colon, blood, breast, liver, and pacritaxel-resistant lung(A549-Pa) cancer cells to evaluate the cancer cell growth inhibition. From the screening, derivatives of halichondramide, wondonin, and psammaplin A, F452-R(Asperphenin), Salternamide A, etc were selected as Hit compounds. Their anti-cancer abilities were tested by cell signaling system evaluation and animal experiment.
2. The anti-angiogenesis activity and its mechanism of were identified after the treatment of 6'-debromohamacanthin A, separated from marine organism Sponge Spongosorites sp., on HUVEC endothelial cells and endothelium induced from mouse embryonic stem cells. Also the anti-angiogensis ability of Wondonin derivatives which is selected as lead compound was confirmed using various in vitro experiments to figure out the intracellular mechanism and in vivo experiments in zebrafish animal model. Such research verified superior ability of wondonin derivatives to inhibit the new blood vessel formation.
3. SP0505-C, D, VJP258-A, D, E, F were selected because of their anti-proliferative ability, and how they suppress the cell growth was examined. The pathway of SP0505-C to induce apoptosis in lung cancer cell line(A549), colorectal cancer cell line(HCT116), and hepatic cancer cell line(SK-HEP-1) was identified. VJP258-A showed anti-proliferative ability in lung cancer cell line(A549), hepatic cancer cell line(SK-HEP-1) via G2/M arrest. However, VJP258-A suppressed the cell growth in pacritaxel-resistant lung cancer cell line(A549-Pa) via apopotosis. VJP259-F displayed anti-proliferative ability by causing G2/M arrest. Especially 19Z-Halichondramide separated from sponge Chondrosia corticata suppress lung cancer growth by controlling the mTOR signaling system and blocking the G2/M cell cycle progression.
4. The compounds that have ability to control the transcriptional activity of TCF/LEF transcription factor, which binds with β-catenin, were selected from the screening. From the research, 03CH5, SP0505, and several compounds affiliated with VJP258 are found to be active in controlling the luciferase activity. The research was conducted to demonstrate the anti-cancer activity and the mechanism of Saltermamide A in colorectal cancer. The result showed Saltermamide A inhibits HIF-1α, an important factor of angiogenesis, and is related to various cell signaling pathways (PI3K/Akt, MAPKs, p53).
5. Psammaplin A derivative was chosen to be a lead compound so the compound was synthesized to investigate the anti-cancer ability. The experiments in the cellular level showed prominent anti-cancer ability. When the compound was tested to check the in vivo effectiveness using an animal model, psammaplin A derivative showed apparent tumor suppressing property in xenograft model (When it was injected at 30mg/kg, the tumor growth was suppressed more than 47% compared to control). The anit-cancer ability of F452-R was proved by animal testing (When it was injected at 8mg/kg, the tumor growth was suppressed more than 67%). The detailed mechanism was investigated. Another research was conducted to demonstrate anti-angiogenesis effect of salternamide A by inhibiting HIF-1α, an important factor for angiogenesis.
6. About 1,200 kinds of marine natural products, extracts, and their synthetic derivatives were evaluated for their inhibitory activities against several representative pathogenic microorganisms including G( ) and G(-) bacteria, MRSA, fungi and antimicrobial target enzymes (SrtA, ICL, Na/ K -ATPase and actin). From the screening, 65 compounds were selected as potent bioactive (hit) compounds. Among them, psammaplin A synthetic derivatives (PSA 3041, 3031, 3051, 3261, 3101, 3271) showed potent inhibitory activity against MRSA, Several active compounds such as bis(indole) alkaloids, beta-carboline alkaloids and halichondramides exhibited potent antibacterial and antifungal activities.
7. Psammaplin A derivative was chosen to be a lead compound so the compound was synthesized to investigate the antibacterial activity toward MRSA. The experiments in the cellular level showed excellent anti-MRSA activity. Especially, two psammaplin A synthetic derivatives (PSA3261 and PSA3271) showed notable antibacterial and anti-MRSA activities. When the compound was tested to check the in vivo effectiveness using an animal model, these showed apparent antibacterial activity in mouse model (Balb/c female mouse) infected with Staphylococcus aureus Newman strain (When it was injected at 10 mg/kg, the survival rate was 100% compared to control). These two compounds are selected as lead compounds for the next study.
8. In addition, various compounds were identified as potent enzyme inhibitors against SrtA (aptamines, psammaplin A synthetic derivatives), ICL (cardiolide E, bahamaolide), Na/ K-ATPase (PSA3105, 416H, SNU607-B, SD9-A) and actin depolymerization (halichondramides). From the screening, several bioactive compounds were selected for the further study.
9. After the development of the HPLC methods for halichondramide and psammaplin A, their physico-chemical properties, i.e. solubility, lipophilicity, and permeability were evaluated using the analytical methods. The methods for the determination of psammaplin A and its derivatives in rat plasma were validated in terms of selectivity, linearity, precision, accuracy, stability, extraction recovery, and matrix effect according to the FDA guideline. The methods were then applied to the pharmacokinetic studies of psammaplin A and its derivatives in rats and the pharmacokinetic parameters, including total area under the plasma concentration– time curve (AUC), terminal half-life (t_1/2), time-averaged total body clearance (CL), apparent volume of distribution at steady state (V_ss),and mean residence time (MRT) were calculated. The pharmacokinetic superiority of the synthetic derivatives over the original psammaplin A were proved by conducting the statistical analyses of the pharmacokinetic parameters.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 보고서 요약서 ... 3
• 요약문 ... 4
• SUMMARY ... 9
• CONTENTS ... 15
• 목차 ... 16
• List of Figures ... 17
• List of Tables ... 22
• 제 1 장 연구개발과제의 개요 ... 25
• 제 1 절 목 적 ... 25
• 제 2 절 필요성 ... 26
• 제 2 장 국내외 기술개발 현황 ... 30
• 제 1 절 국외 현황 ... 30
• 제 2 절 국내 현황 ... 31
• 제 3 장 연구개발수행 내용 및 결과 ... 35
• 제 1 절 연구 목표 및 평가의 착안점 ... 35
• 제 2 절 연구내용 및 방법 ... 38
• 제 3 절 연구 결과 ... 55
• 제 3 절 연구 성과 ... 268
• 제 4 장 목표달성도 및 관련분야에의 기여도 ... 297
• 제 1 절 목표달성도 ... 297
• 제 2 절 관련분야에의 기여도 ... 303
• 제 5 장 연구개발결과의 활용계획 ... 304
• 제 1 절 추가연구의 필요성, 타 연구에의 응용 ... 304
• 제 2 절 연구개발결과의 활용방안 및 기대효과 ... 305
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 308
• 제 7 장 연구시설ㆍ장비 현황 ... 311
• 제 8 장 참고문헌 ... 312
• 끝페이지 ... 315