생약제제의 이화학적 품질평가에 관한 연구 갈근탕(葛根湯)의 연구

정지형,김진수,박희준,박종희,Jung, Jee-H.,Kim, Jin-Soo,Park, Hee-Juhn,Park, Jong-Hee 한국생약학회 1997 생약학회지 Vol.28 No.1

The prescription of Gal Gun Tang, which has been used for treatment of cold, fever, and muscular pain in Chinese herb medicine, is produced in the form of decoction However, the storage problem for this dosage form remains unsettled. Using HPLC and GC, we examined quantitative change of major constituents caused by time-progress and temperature-change. Nine major constituents, such as cinnamaldehyde, cinnamic acid, ephedrine, puerarin, paeoniflorin, daidzin, benzoic acid, glycyrrhizin and liquiritin, were selected as references. The content of cinnamaldehyde significantle decreased by the increase of temperature. While that of cinnamic acid increased. Benzoic acid showed the most significant change of the content in three months at $40^{\circ}C$. It is suggested that most of constituents are considerably stable when kept frozen.

인체 혈구암세포 U937에서 해양해면동물에서 추출된 Pectenotoxin-2에 의한 Apoptosis의 유발에 관한 연구

신동역,강호성,배송자,정지형,최영현,Shin, Dong Yeok,Kang, Ho Sung,Bae, Song-Ja,Jung, Jee H.,Choi, Yung Hyun 한국해양바이오학회 2006 한국해양바이오학회지 Vol.1 No.2

본 연구에서는 U937 인체 백혈병 세포의 증식에 미치는 PTX-2의 영향을 조사한 결과, PTX-2의 처리에 따라 U937 세포는 처리 농도 및 처리 시간 의존적으로 심한 형태적 변형과 함께 증식이 억제되었다. 이러한 PTX-2 처리에 의한 U937 세포의 증식억제는 apoptosis 유발과 관련이 있었으며, 이를 DAPI staining에 의한 apoptotic body 형성, flow cytometry를 이용한 sub-G1 세포 빈도의 정량적 분석을 통하여 확인하였다. 이러한 PTX-2 처리에 의한 U937 세포의 apoptosis 유발은 Bcl-2 family에 속하는 anti-apoptotic 인자인 Bcl-$X_L$의 발현 감소 및 IAPs family에 속하는 유전자들의 선택적 발현 감소와 연관성이 있음을 알 수 있었다. 이상의 결과들은 인체 암세포에서 PTX-2의 항암작용을 이해하는데 중요한 자료가 될 것이고 나아가 PTX-2을 포함한 그와 유사한 항암제 후보물질들의 연구에 있어서 기초 자료로서 사용될 수 있을 것으로 생각된다. Natural product compounds are the source of numerous therapeutic agents. The marine environment produces natural products from a variety of structural classes exhibiting activity against numerous disease targets including anticancer agents. Among these, pectenotoxin-2 (PTX-2), which was first identified as a cytotoxic entity in marine sponges, which depolymerizes actin filaments, was found to be highly effective and more potent to activate an intrinsic pathway of apoptosis in p53-deficient tumor cells compared to those with functional p53 both in vitro and in vivo. However, the anti-proliferative mechanism of the compound at non-cytotoxic concentrations has not yet been explored. In the current study, we sought to investigate anti-proliferation and apoptosis of PTX-2 against U937 human leukemic cells and its underlying molecular mechanism. Exposure of U937 cells to PTX-2 resulted in growth inhibition and induction of apoptosis in dose- and time-dependent manner as measured by MTT assay, fluorescent microscopy and flow cytometric analysis. The anti-proliferative effect of PTX-2 was associated with a marked increase in the expression of cyclin-dependent kinase p21 (WAF1/CIP1) mRNA which was tumor suppressor p53-independent. The increase in apoptosis was connected with a time-dependent down-regulation of anti-apoptotic Bcl-XL and inhibitor of apoptosis proteins (IAPs) family such as XIAP and cIAP-2. Though additional studies are needed, these findings suggested that PTX-2-induced inhibition of U937 cells was associated with the induction of apoptotic cell death and the results provided important new insights into the possible molecular mechanisms of the anti-cancer activity of PTX-2.

진세노사이드 Rh2의 방향선택적 합성

신명희(Myoung Hee Shin),정지형(Jee H. Jung),장은하(Eun Ha Chang),임광식(Kwang Sik Im) 대한약학회 2001 약학회지 Vol.45 No.4

Ginsenoside Rh2, a minor glycoside constituent of the red ginseng is known as an unique antitumor compound. Several attempts to prepare it in a large scale including semisynthesis from bettalafolientriol, an 3-epimer of 20(S)-protopanaxadiol, has been reported. We have previously reported a synthesis of ginsenoside Rh2 from 20(S)-protopanaxadiol obtained by alkaline hydrolysis of total ginsenoside. The regioselective synthesis of this compound was achieved by protection of 12-OH group.

국내산 해면 Spirastrella abata로부터 Sterol Peroxide 유도체의 분리

임광식,남경인,심정자,정지형,Im, Kwang-Sik,Nam, Kyung-In,Sim, Chung-J.,Jung, Jee-H. 한국생약학회 2000 생약학회지 Vol.31 No.4

Marine sponges are known to be a source of diverse sterols. In our study on the cytotoxic components of the marine sponge Spirastrella abata, $5{\alpha},8{\alpha}-epidioxy\;{\Delta}^6$ sterols (1-5) and $5{\alpha},8{\alpha}-epidioxy\;{\Delta}^{6,9(11)}$ sterols (6-7) were isolated. The structures were identified based on the analyses of $^1H-NMR,\;^{13)C-NMR$, and MS data. These compounds were assayed for cytotoxicity against 5 human solid tumor cell lines including A549, SK-OV- 3, SK-MEL-2, XF498, and HCT15.

해양미세조류의 라디칼소거활성 검색

최진석,이원갑,김동수,최홍대,최재수,정지형,임광식,최원철,손병화,Choi, Jin-Seok,Lee, Won-Kap,Kim, Dong-Soo,Choi, Hong-Dae,Choi, Jae-Sue,Jung, Jee H.,Im, Kwang-Sik,Choi, Won-Chul,Son, Byeng-Wha 한국생약학회 2000 생약학회지 Vol.31 No.2

In order to screen new radical scavenging principle which is expected to be antiaging drug lead, we have investigated 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of the marine microalgae, greenalgae(10 speices), diatom (10 speices) and blue-green algae (10 speices). The significant activities$(IC_{50}:\;<100\;{mu}g/ml)$ were observed in 4 species of green algae (MA002, 006, 009, 010), 1 species of diatom (MA015) and 5 species of blue-green algae (MA017, 018, 019, 024, 025). Within the scope of family tested, MA009 $(IC_{50}:\;=78\;{mu}g/ml)$, MA015 $(IC_{50}:\;=38\;{mu}g/ml)$ and MA019 $(IC_{50}:\;=41\;{mu}g/ml)$ displayed the most significant activity. Among the marine microalgae tested at family level, cyanophycean blue-green algae was shown to be the most active family on screening of new bioactive compounds.

종양억제유전자 p53 결손 인체간암세포에서 Pectenotoxin-2에 의한 Apoptosis 유도

신동역(Dong Yeok Shin),김기영(Gi Young Kim),최병태(Byung Tae Choi),강호성(Ho Sung Kang),정지형(Jee H. Jung),최영현(Yung Hyun Choi) 한국생명과학회 2007 생명과학회지 Vol.17 No.10

해양생물 유래 항암활성을 가지는 천연물의 탐색과정에서 해면동물에서 유래된 PTX-2는 p53 결손 암세포에서 세포독성 효과가 높은 것으로 보고된 바 있다. 본 연구에서는 인체 간암세포 모델을 이용하여 PTX-2의 효능을 조사한 결과는 p53 결손 Hep3B 세포에서 p53 정상 HepG2에 비하여 항암활성이 매우 높았으며, 이는 apoptosis 유발과 연관성이 있음을 확인하였다. PTX-2에 의한 Hep3B 세포의 apoptosis 유발은 DFF family의 발현 변화, pro-apoptotic Bax 및 Bcl-xS 단백질의 발현 증가, caspases (-3, -8 및 -9)의 활성화 등이 관여함을 알 수 있었다. PTX-2는 또한 Hep3B 세포에서 AKT 및 ERK1/2의 활성화를 유도하였으며, caspase-3, AKT 및 ERK1/2의 특이적 저해제에 의하여 PTX-2에 의한 세포증식 억제 효능이 유의적으로 감소되었다. 본 연구는 PTX-2에 의한 Hep3B 세포에서의 apoptosis 유도에 AKT 및 ERK1/2 신호 전달 경로가 중요한 역할을 하고 있음을 보여주는 결과이다. Through the screening of marine natural compounds that inhibit cancer cell proliferation, we previously reported that pectenotoxin-2 (PTX-2) isolated from marine sponges exhibits selective cytotoxicity against several cell lines in p53-deficient tumor cells compared to those with functional p53. However, the molecular mechanisms of its anti-proliferative action on malignant cell growth are not completely known. To further explore the mechanisms of its anti-cancer activity and to test whether the status of p53 in liver cancer cells correlates with their chemosensitivities to PTX-2, we used two well-known hepatocarcinoma cell lines, p53-deficient Hep3B and p53-wild type HepG2. We have demonstrated that PTX-2 markedly inhibits Hep3B cell growth and induces apoptosis whereas HepG2 cells are much more resistant to PTX-2 suggesting that PTX-2 seems to act by p53-independent cytotoxic mechanism. The apoptosis induced by PTX-2 in Hep3B cells was associated with the modulation of DNA fragmentation factor (DFF) family proteins, up-regulation of pro-apoptotic Bcl-2 family members such as Bax and Bcl-xS and activation of caspases (caspase-3, -8 and -9). Blockade of the caspase-3 activity by caspase-3 inhibitor, z-DEVD-fmk, prevented the PTX-2-induced growth inhibition in Hep3B cells. Moreover, treatment with PTX-2 also induced phosphorylation of AKT and extracellular-signal regulating kinase (ERK), but not c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MARK). Specific inhibitors of PI3K inhibitor (LY294002) and ERK1/2 inhibitor (PD98059) significantly blocks PTX-2-induced-anti-proliferative effects, whereas a JNK inhibitor (SP600125) and a p38 MAPK inhibitor (SB203580) have no significant effects demonstrating that the pro-apoptotic effect of PTX-2 mediated through activation of AKT and ERK signal pathway in Hep3B cells.