가상 검색 및 시험관 시험을 이용한 총명탕 중 주성분들에 대한 약물작용 및 대사 예측

권영이,Kwon, Young-Ee 대한약학회 2007 약학회지 Vol.51 No.6

Chong Myung Tang is consisted of three medicinal herbs (Acori Graminei Rhizoma, Polygalae Radix and Hoelen cum Radix). It has been used as a medicine for the purpose of learning and memory improvement. In this paper, Chong Myung Tang was screened the biological activities for Alzheimer's disease. The extract (70% ethanol) of Acari Graminei Rhizoma (1 mg/ml) showed that acetylcholinesterase (AChE) and amyloid beta ($A{\beta}$) peptide aggregation inhibitory potency are 43.1% and 76.5%, respectively. The extract of Polygalae Radix showed inhibitory activity against $A{\beta}_{1-42}$ peptide aggregation (51.5%). To predict the drug-likeness, oral absorption ability; blood-brain barrier (BBB) penetraion rate, mutagenecity and carcinogenicity; in silico screening was performed against 16 compounds in the three medicinal herbs. According to the results, all compounds have appropriate chemical structures as medicines. The six compounds in Acori Graminei Rhizoma and the five compounds in Hoelen cum Radix showed excellent oral absorption rate and BBB penetration rate. The four compounds in Polygalae Radix showed excellent oral absorption rate, but their BBB penetration was presented low rate. And, the extract of Hoelen cum Radix didn't show AChE and $A{\beta}_{1-42}$ peptide aggregation inhibitory activities in vitro. Therefore, their activity in brain may be other mechanism. According to all of the results, in silico prediction technology is convenient and effective to determine biological active compounds in medicinal herbs.

베타아밀로이드응집 억제제 도출을 위한 새로운 벤질피페리디닐에터 유도체의 합성

권영이,Kwon, Young-Ee 대한약학회 2006 약학회지 Vol.50 No.5

We designed and synthesized new benzylpiperidinyl ether derivatives as beta-amyloid aggregation inhibitors for the development of novel anti-Alzheimer's disease agents. As starting material, 4-hydroxypiperidine was used. For protection of the amine group in piperidine (2), di-tert-butyl dicarbonate was reacted with 4-hydroxypiperidine in the presence of triethylamine. For introduction of benzyl group, benzylbromide was treated with compound 2 in dioxane. After deprotection of Boc group on amine in compound 3, ester (5) was synthesized by addition of ethyl-4-chlorobutyrate. The alcohol 6 was synthesized by hydride reduction of 5 using $LiAlH_4$. To obtain final products (7-14), the alcohol 6 was condensed with each of substituted benzoic acids. To screen beta-amyloid aggregation inhibition of the products, thioflavinT assay was performed using $A{\beta}1-42\;at\;37^{\circ}C$ for 26 h incubation, in vitro. From the result of screening, compound 8, 9, 11 and 12 showed effective activity about $65{\sim}85\;{\mu}M\;as\;IC_{50}$ value. Among the prepared compounds, 4-[4-(benzyloxy)piperidino]butyl-4-chlorobenzoate (8) was the most effective inhibitor having $IC_{50}\;of\;65.4{\mu}M$.

Response of Metastatic Cancer Cells to Thermal Changes in vitro

안상길,권영이,최호순,권중균,유진영,김종룡,김원규,Ahn, San-Gil,Kwon, Young-Ee,Choi, Ho-Soon,Kwon, Jung-Kyun,Yoo, Jin-Young,Kim, Jong-Ryong,Kim, Won-Kyu Korean Society of Electron Microscopy 2007 Applied microscopy Vol.37 No.4

암세포가 있는 장소의 온도를 변화시키는 것은 하나의 암 치료 방법이 될 수 있다. 명확한 기전은 아직 잘 밝혀져 있지 않지만, 고온은 미토콘드리아로 신호를 전달해서 cytochrome c를 분비시키는 세포자멸사로의 길로 유도하는 것으로 알려져 있다. 저온은 $30^{\circ}C$ 미만에서 세포자멸사를 유도하지만 심하지 않은 저온에서는($35{\sim}33^{\circ}C$ 혹은 $31{\sim}29^{\circ}C$)오히려 세포자멸사를 막는 것으로 알려져 있다. CC-t6와 GB-d1세포 주는 림프절로 전이된 사람의 담관암과 담낭암에서 확립한 것으로, 이와 같은 전이성 암세포가 온도 변화에 어떻게 반응을 하는지를 연구하기 위해 고온노출($37{\rightarrow}43^{\circ}C$)과 저온노출($37{\rightarrow}17.4^{\circ}C$)을 시행하였다. 세포의 종류나 온도 변화를 통한 스트레스의 방법과 관계없이 죽는 세포가 관찰되었으며, 고온노출이 가장 심한 영향을 주었다. 이런 죽어가는 세포는 세포자멸사가 아닌 세포괴사의 경로를 거치고 있었다. 투과전자현미경을 이용한 관찰에서 세포자멸사적인 모습은 보이지 않았고, caspase-3, -9, cytochrome c, Bax 같은 세포자멸사와 관련된 단백질의 변화도 관찰되지 않았고, 열충격단백질 70과 27도 증가하였다. 결국 CC-t6와 GB-d1 세포는 온도변화를 통한 스트레스를 주었을 경우 세포괴사로 죽음을 알 수 있었다. 온도변화를 통한 스트레스는 열충격단백질의 증가와 함께 세포괴사를 일으켰다. GB-d1과 CC-t6 세포에서 고온은 가장 심각하게 세포괴사를 일으켰으며, 저온은 초기에는 세포괴사를 유발하였으나 12시간 경과후에는 세포분열이 더욱 활발하게 일어나 세포의 생명력을 연장시켜주었다. 결국 이 실험에서는 전이성 암세포를 제거하는 방법으로는 고은이 가장 효과적이며 유용함을 알 수 있었다. Alteration of temperature is one of cancer therapies. In general, severe hyperthermia(around $43^{\circ}C$) and hypothermia(around $18^{\circ}C$) trigger apoptosis through mitochondria, though the specific mechanism is still unknown. CC-t6 and GB-d1 cell lines, which were originally derived from human cholangiocarcinoma and gall bladder cancer, were established from a metastatic lymph node. To investigate the mechanism of metastatic cancer cell response to thermal stresses, hyperthermia($37^{\circ}C{\rightarrow}43^{\circ}C$) and hypothermia($37^{\circ}C{\rightarrow}17.4^{\circ}C$) were designed. Thermal stresses did not induce apoptosis but necrotic cell death. Any alterations of caspase-3, -9, cytochrome c, Bax, and Bcl-2 were not found in both hyperthermia and hypothermia exposed fells using western blot analysis. In the transmission electron microscopy, typical necrotic, but not apoptotic, changes were observed. These results suggest that temperature changes induce cell death through necrotic pathway in metastatic cancer in vitro, and it can be one of effective anticancer methods.

고속액체크로마토그라피법을 이용한 사람 혈장 중 알리벤돌(Alibendol)의 정량 및 검증

송현호(Hyun Ho Song),유지영(Ji-young Yu),김보겸(Bo Gyeom Kim),박현주(Hyeon Ju Park),최광식(Kwang Sik Choi),권영이(Young Ee Kwon) 大韓藥學會 2010 약학회지 Vol.54 No.4

The aim of this study was to develop and validate for determination of alibendol in human plasma by HPLC method. After precipitation of 500 μl plasma samples by 50% methanol 50 μl and 60% perchloric acid 30 μl and the supernatant 50 μl was injected into HPLC. The assay was performed isocratically using 10 mM potassium phosphate (pH 3.0) and acetonitrile (80 : 20, v/v) as mobile phase. The C18 column (particle size 3.5 μm, 4.6×50 mm, Zorbax Eclipse) was used as a solid phase. The mobile phase was delivered at a flow-rate of 1.7 ml/min, detection was by ultraviolet absorption at 232nm and concentrations were calculated on the basis of peak areas. In these conditions, alibendol can be separated from ethylparaben, the internal standard, and endogenous substances. The retention times of alibendol and ethylparaben were just about 2.6 and 3.5 minutes, respectively. This rapid HPLC method was validated by examining the precision and accuracy for inter- and intra-day analysis. The standard curve was linear (R2=1.0000) over the concentration range of 0.05~20 μg/ml. The inter-day relative standard deviation (R.S.D.) and accuracy were 0.2~12.2% and 94.4~101.2% (82.7% at the lower limit of quatitation). The intra-day R.S.D. and accuracy were 0.1~11.8% and 98.8~102.5%, respectively. The method was successfully applied to the determination of alibendol in plasma for a pharmacokinetic study.

붉은 서나물 잎 (Erechitites hieracifolia Raf.)에서의 Polyphenoloxidase 활성측정 및 항산화효오 특성분석

김안근(Kim An Keun),이상은(Lee Sang Eun),김국환(Kim Kuk Hwan),권영이(Kwon Young Ee) 대한약학회 2002 약학회지 Vol.46 No.4

Polyphenoloxidase activity (PPO) in the leaves of Erechitites hieracifolia was estimated by Warburg's manometric method. The emzyme was most reactive toward chlorogenic acid followed by caffeic acid. Biethyldithiocarbamate and potassium cyanide were shown powerful inhibition rate to the polyphenoloxidase from the leaves of Erechitites hieracifolia . We confirme d antioxidant activity of the leave s ofErechitites hieracifolia by DPPH (1,1- diphenyl -2-picrylhydrazyl) radical scavenging method. Electrophorectic isoenzyme banding patterns of superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) were observed by native PAGE. The correlation of PPO and antioxidant enzymes is not investigated yet. That is need to further study.

모발생장기 유도 C3H생쥐에 있어서 미녹시딜과 생약추출 혼합 조성물의 모발 재성장 유도 효능

이계호(Kye Ho Lee),한선일(Sun Il Han),박길홍(Gil Hong Park),권영이(Young Ee Kwon) 대한약학회 2003 약학회지 Vol.47 No.1

The hair cycle consists of three phases, growth (anagen), involution (catagen) and quiescence (telogen) phases. In order to evaluate hair re-growth effect of herbal extracts mixture containing the 70% ethanol extracts of Polygoni Mulitiflori Radix, Mori Cortex Radix, Gingo Biloba Folium and Pine bud, we have examined the induction of the anagen phase and elongation of the anagen period using C3H mice. MorPhological examination was done by Hattori and Ogawa's method. Enzyme activites of γ-glutamyl transpeptidase (γ-GT) and alkaline phosphatase(ALP) was detected by Bessey-Lovry-Brock's method. Enzyme activity as a biochemical marker of cycle was investigated in the third hair cycle period of C3H mice after depilation. 3% Milnoxldll treated group and herbal extract mixture treated group were shown 3 days earlier initiation of anagen than control group. In cycling mousse skin, γ-GT activity is pronounced during anagen and greatly diminished during telogen. Herual extract mixture has shown promising hair re-growth efect on hair follicula cycles of C3H mice.

리보플라빈을 가진 수용성 백금(Ⅱ)착체의 합성과 항암활성 및 세포주기와의 관계

황규자,권영이 숙명여자대학교 약학연구소 1999 약학논문집-숙명여자대학교 Vol.16 No.-

A series of vitamine-containing Pt(II) complexes of the type [Pt(Rf)(L)], (where Rf=riboflavin, L=ethylenediamine, 1,3-propanediamine, 1,4-butanediamine) was synthesized and characterized by IR, electronic absorption, elemental analysis and FAB-Mass. The coordination sites of Rf to Pt(II) ions were determined to be N(5) and 0(6) with resultant chelate ring formation. These compouds have much better water solubility(145- 150mg/ml) than cisplatin has(lmg/ml). The anticancer activity of this Riboflavin-containing Pt(II) series was investigated by MTT assay against mouse and human leukemia cell lines in vitro. Interaction between Pt(II) complex and DNA were analyzed by flowcytometry, and showed G2/M arrest in LI210 cell line.

7원환 구조를 갖는 새로운 백금착체의 합성에 대한 연구

황규자,권영이 숙명여자대학교 약학연구소 1998 약학논문집-숙명여자대학교 Vol.14 No.-

Platinum complexes having 7 .membered ring structure with Riboflavin(RF) and Flavin mononucleotide(FMN) as leaving group were synthesized to enhance antitumor activity and solubility. Platinum complexes with RF and FMN were determined by ir spectra, nmr spectra, mass and elemental analysis.

1,4-Butanediamines를 carrier ligand로 하는 7원환 백금착체의 설계 및 합성

황규자,이근임,권영이 숙명여자대학교 약학연구소 1995 약학논문집-숙명여자대학교 Vol.11 No.-

Platinum complexes having 7-membered ring structure with N,N'-bis(2-hy-droxyethyl)butylenediamine(HeBn) and N,N'-bis(2-chloroethyl)butyelenediamine(CeBn) as carrier ligands were synthesized to enhance antitumor activity and to decrease the toxicity. Platinum complexes were prepared by reacting K_2PtI_4 with HeBn or CeBn. IR, NMR, elementary analysis, ICP-AES and titration method were applied for identification and analysis of the synthesized platinum complexes.

새로운 백금착체의 설계와 그 항암활성

황규자,이근임,권영이 숙명여자대학교 약학연구소 1996 약학논문집-숙명여자대학교 Vol.12 No.-

Pt(Ⅱ) and Pt(Ⅳ) complexes having six-membered chelate rings with N,N'-bis(2-chloroethyl)propanediamine (2-CePn) as carrier ligand have been prepared and characterized by IR and ^1H NMR. These complexes were examined for their in vitro cytotoxicity against leukemia L1210 and P388 cell lines. The cytotoxicity of platinum complexes with 1,3-propanediamine were improved by introduction of 2-chloroethyl group which nitrogen mustard, and alkylating agent, contains. The Pt(Ⅱ) complex showed more effective cytotoxicity than Pt(Ⅳ) complex.