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송윤경,진선경,한의식,안미령,정주연,이이다,조일영,김동섭,지은희,박효영,오정미,신원,이선희,김인규,Song, Yun-Kyoung,Jin, Sun-Kyung,Han, Eui-Sik,Ahn, Mee-Ryung,Jung, Ju-Yeon,Lee, Rhee-Da,Cho, Il-Yong,Kim, Dong-Sub,Ji, Eun-Hee,Park, Hyo-Young,Oh, 대한약학회 2011 약학회지 Vol.55 No.4
Gastritis is the most common disease among Korean. The demand for the development of gastritis drugs has been increasing. Currently, however, there is no guideline available for the clinical evaluation of gastritis drugs worldwide. As a consequence, domestic and international pharmaceutical companies make errors in the drug development processes, and it becomes difficult for them to establish the scientific validity and objectivity of newly developed drugs. The objective of this study was to develop the Guideline for Clinical Trials Evaluation of Gastritis which can be used in improving the quality and consistency of clinical trials. First, we collected and reviewed the clinical trials on gastritis drugs that were available from Japan Pharmaceuticals and Medical Devices Agency and Korea Food and Drug Administration (KFDA), and investigated the recent research trends on clinical trials of gastritis drugs. Reviewers from KFDA and National Institute of Food and Drug Safety Evaluation and scientific experts from the pharmaceutical industries developed the guidelines through regularly scheduled meetings. Opinions and consultation from academic fields and industry experts were also obtained. This project will provide the clinical trial practitioners, investigator and reviewers the scientific and rational guidelines for performance and evaluation of clinical trials for gastritis drugs. Furthermore, we hope this guideline contributes to establishing the national competitiveness, improving the quality of clinical trial, and encouraging researches on drug development for gastritis.
천연감미료 스테비오사이드와 스테비올의 생체내, 시험관내 유전독성평가
오혜영(Hye Young Oh),한의식(Eui Sik Han),최돈웅(Don Woong Choi),김종원(Jong Won Kim),손수정(Soo Jung Son),엄미옥(Mi Ok Eom),강일현(Il Hyun Kang),강혁준(Hyuck Joon Kang),하광원(Kwang Won Ha) 대한약학회 1999 약학회지 Vol.43 No.5
The standard operation procedure of mouse lymphoma L5178Y tk+/- -3.7.2C gene mutation assay (MOLY) has been regarded as a sensitive In vitro mammalian cell gene mutation assay that is capable of detecting clastogens as well as mutagens. Using MOLY, one of natural sweetner, stevioside (5mg/ml) and its aglycon, steviol (340mcg/ml) were evaluated the mutagenicity. Stevioside and steviol did not induce mutagenicity in MOLY. On the other hand, stevioside (250mg/kg, B.W.) and steviol (200mg/kg, B.W.) were also evaluated their ability to induce mucronuclei in regenerating hepatocytes and bone marrow cells of ddY mice. From these results, stevioside and steviol did not induce any mutagenic effect both MOLY and In vivo micronucleus test.
Syrian hamster embryo 세포와 mouse embryo BalB/c 3T3 세포에서의 bisphenol A의 세포 형질전환 연구
김종원(Jong Won Kim),한의식(Eui Sik Han),박미선(Mi Sun Park),엄미옥(Mi Ok Eom),전혜승(Hye Seung Jun),민수진(Su Jin Min),김인숙(In Sook Kim),정해관(Hai Kwan Jung),심웅섭(Woong Seop Sim),오혜영(Hye Young Oh) 한국환경성돌연변이발암원학회 2001 한국환경성돌연변이·발암원학회지 Vol.21 No.1
To identify nongenotoxic carcinogen determined as negative by ICH guideline-recommended standard genotoxicity test battery; Ames test, chromosome aberrration assay, mouse lymphoma tk+/- assay, in vivo micronucleus assay, we picked bisphenol A as a model compound. In this study, we applied in vitro BalB/c 3T3 cell transformation assay and Syrian hamster embryo (SHE) cell transformation assay. Bisphenol A was treated upto 769.2 ug/ml in BalB/c 3T3 cells and upto 125 ug/ml in SHE cells. bisphenol A didn't induced morphological transformation both with one stage treatment protocol and with two stage treatement protocol. But, treated for 48 hr, Bisphenol A induced morphological transformation significantly in SHE cells.
Di(2-ethylhexyl) phthalate에 의해 유도된 DNA 손상과 소핵 형성
김종원(Jong Won Kim),한의식(Eui Sik Han),박미선(Hai Kwan Jung),엄미옥(Mi Sun Park),김인숙(Mi Ok Eom),전혜승(In Sook Kim),정해관(Hye Seung Jun),심웅섭(Woong Seop Sim),오혜영(Hye Young Oh) 한국환경성돌연변이발암원학회 2001 한국환경성돌연변이·발암원학회지 Vol.21 No.1
Di-2-ethylhexyl phthalate (DEHP) is the most commonly used phthalate ester in polyvinyl chloride formulations including food packing and storage of human blood. DEHP is a well known as non-genotoxic carcinogen and endocrine disrupting chemical (EDC). DEHP have shown all negative results in ICH-guildeline recommended standard genotoxicity test battery. In this study, to assess the clastogenic and DNA damaging effect in human-derived tissue specific cells, DEHP was treated in human derived MCF-7 cells, HepG2 cells, LNCap cells, BeWo cells, MCF-10A cells, and female peripheral blood cells using micronucleus assay and in human<br/> breast carcinoma MCF-7 cells up to 1.28×10-²M using Comet assay. The in vitro micronucleus assay is a mutagenicity test system for the detection of chemicals which induce the formation of small membrane bound DNA fragment i.e. micronuclei in the cytoplasm of interphase cells, originated from clastogenic and/or aneugenic<br/> mechanism. The single cell gel electrophoresis assay (Comet assay) is used to detect DNA strand-breaks and alkaline labile site. In our results, DEHP increased significantly and/or dose-depentently and time-dependently micronucleus frequency at the 6 and 24 hr without metabolic activation system only in MCF-7 cells. DEHP treated with 2 hrs in MCF-7 cells using Comet assay induced DNA damage dose-depentantly.
사람 암세포에서의 O6-methylguanine-DNA methyltransferase 의 발현과 알킬화 항암제에 대한 감수성
오혜영(Hye Young Oh),정해관(Hae Kwan Jung),한의식(Eui Sik Han),정성철(Sung Chul Jung),허옥순(Ok Soon Heo),손수정(Soo Jung Sohn),김영미(Young Mi Kim),홍성렬(Sung Youl Hong),이향우(Hyang Woo Lee),하광원(Kwang Won Ha) 한국응용약물학회 1995 Biomolecules & Therapeutics(구 응용약물학회지) Vol.3 No.2
Five human cancer cell lines (HeLa S3, Hep 3B, KATO III, Hs 683, HeLa MR) and one human normal cell line (WI-38) were examined cell viability, northern blot analysis, western blot analysis, and in situ hybridization for the expression of O^6-methylguanine-DNA methyltransferase (MGMT), which can repair O^6-methylguanine produced in DNA by alkylating agents. In cell viability test, the lethal sensitivities of each strain against anti-tumor drug N,N-bis(2-chloroethyl)- N-nitrosourea (BCNU) were counted, and both BCNU treated and untreated cell extracts were examined for their MGMT inducibility by RNA dot blot analysis. Cell lines did not show MGMT induction by BCNU pretreatment. The MGMT activity was assayed by measuring the ³H radioactivity transferred from the substrate DNA containing [methyl-³H]-O^6-methylguanine to acceptor molecules in the cell extracts. Extracts from the majority of tumor strains and normal cells contained substantial MGMT activity of varying degree, while the known Mer^- cell (lacked or severely depleted in MGMT activity) Hela MR, and Hs 683 (proved to be Mer^-) were much more sensitive to BCNU than the rest of tumor strains, as measured by cell viability test. Overall results above, KATO III showed the highest expression level of MGMT among the strains examined. Furthermore, with all the tumor and normal strains tested, a good correlation was observed between MGMT expression and cellular resistance to BCNU. The varying levels of expression of MGMT in human cancer cells found in this study should provide a molecular basis for MGMT expression among tumor strains from different tissue origin, the information of antitumor agents selection for chemotherapy of cancers.
OECD 급성경구투여독성시험 지침의 국내 확립 및 검증
조영래 ( Young Rae Cho ),염영나 ( Young Na Yum ),한의식 ( Eui Sik Han ),곽승준 ( Seung Jun Kwack ),김형섭 ( Hyung Sub Kim ),강미선 ( Mi Sun Kang ),이진영 ( Jin Young Lee ),오재호 ( Jae Ho Oh ),임채형 ( Chae Hyung Lim ),김대성 ( Da 한국동물실험대체법학회 2008 동물실험대체법학회지 Vol.2 No.2
As the study about the non-animal tests came to be internationally active and the interest in the animal welfare gradually became increased, OECD TG (Test Guideline) 401 that has been used since 1987 was abolished in 2002. Because TG401 is acute oral toxicity method depending on survival and death of many animals, it was heavily criticized. Therefore, the three alternative methods were developed. OECD TG420 and TG423 determine the class of chemicals according to GHS (Globally Harmonized Classification System for Chemical Substances and Mixtures) classification and OECD TG425 suggests the predicted LD50 of chemicals using AOT425 program. In this study, 10 chemicals were selected. The internationally admitted TG420, TG423 and TG425 were introduced and established through the method that these chemicals were orally administrated to SD female rats and then, the results were observed. Each chemical belonged to already known GHS class in the study using TG420 and TG423 and predicted LD50 was same or higher in the study using TG425 compared to already known LD50 value. In conclusion, the result of our study confirmed the decrease in the animal number and validated. The international harmonization of the non-animal tests will be pursued through this validation study.
이진영 ( Jin Young Lee ),강미선 ( Mi Sun Kang ),김지명 ( Ji Myoung Kim ),곽승준 ( Seung Jun Kwack ),한의식 ( Eui Sik Han ),한범석 ( Beom Seok Han ),박순희 ( Sue Nie Park ),강태석 ( Tae Seok Kang ),한순영 ( Soon Young Han ) 한국동물실험대체법학회 2010 동물실험대체법학회지 Vol.4 No.1
Repeated dose toxicity testing in rodents is used to have information on potential target organs in terms of toxicity, and the NOAEL (no-observed-adverse-effect-level). ECVAM and ICCVAM have not validated any non-animal methods for assessing chronic toxicity endpoints or repreated exposure target organ toxicity because in vivo systems often poorly resemble in vitro cell culture systems. However, recently ECVAM recommended a proposed approach for the assessment of repeated dose toxicity in vitro. We applied ECVAM`s proposal to assess target organ toxicity using liver and kidney cell lines. In order to predict liver and kidney target organ toxicities, we used three kinds of kidney toxicants(HgCl2, CH3HgCl, CdCl2) and two kinds of liver toxicants(acetaminophen, CuCl2) in human hepatoma(HepG2) cells, human renal tubular epithelial(HK-2) cells and human embryonic kidney(HEK293) cells. We performed cytotoxicity assays, mitochondria damage and calcium influx tests to predict target organ toxicity. Cell viability and cell proliferation were assessed by NRU assay and MTT assays, respectively. Mitochondria potential and cell calcium concentration([Ca2+]i) were estimated by a fluorescence microscopy using JC-1 dye and a fluorometer using Fluo-4/AM dye, respectively. The cytotoxicity of nephrotoxicants(HgCl2, CH3HgCl, CdCl2) from the result of NRU and MTT assay was higher in the kidney cells than in the liver cells. Also, the cytotoxicity of hepatotoxicants(acetaminophen, CuCl2) was higher in the liver cells than in the kidney cells. Mitochondria potential and [Ca2+]i show compatability with cytotoxicity results in the kidney cells but not in the liver cells. The results demonstrated that nephrotoxicants can predict target organ toxicity by in vitro tests like NRU, MTT assay, mitochondria potential and [Ca2+]i. However, hepatotoxicants can predict target organ toxicity by cytotoxicity assays like NRU and MTT assays but not by mitochondria potential and [Ca2+]i.
연구논문 : ECVAM 등의 국제 독성시험법 국내 확립 및 적용
이진영 ( Jin Young Lee ),강미선 ( Mi Sun Kang ),김지명 ( Ji Myoung Kim ),곽승준 ( Seung Jun Kwack ),한의식 ( Eui Sik Han ),한범석 ( Beom Seok Han ),박순희 ( Sue Nie Park ),강태석 ( Tae Seok Kang ),한순영 ( Soon Young Han ) 한국동물실험대체법학회 2010 동물실험대체법학회지 Vol.4 No.1
Repeated dose toxicity testing in rodents is used to have information on potential target organs in terms of toxicity, and the NOAEL (no-observed-adverse-effect-level). ECVAM and ICCVAM have not validated any non-animal methods for assessing chronic toxicity endpoints or repreated exposure target organ toxicity because in vivo systems often poorly resemble in vitro cell culture systems. However, recently ECVAM recommended a proposed approach for the assessment of repeated dose toxicity in vitro. We applied ECVAM`s proposal to assess target organ toxicity using liver and kidney cell lines. In order to predict liver and kidney target organ toxicities, we used three kinds of kidney toxicants(HgCl2, CH3HgCl, CdCl2) and two kinds of liver toxicants(acetaminophen, CuCl2) in human hepatoma(HepG2) cells, human renal tubular epithelial(HK-2) cells and human embryonic kidney(HEK293) cells. We performed cytotoxicity assays, mitochondria damage and calcium influx tests to predict target organ toxicity. Cell viability and cell proliferation were assessed by NRU assay and MTT assays, respectively. Mitochondria potential and cell calcium concentration([Ca2+]i) were estimated by a fluorescence microscopy using JC-1 dye and a fluorometer using Fluo-4/AM dye, respectively. The cytotoxicity of nephrotoxicants(HgCl2, CH3HgCl, CdCl2) from the result of NRU and MTT assay was higher in the kidney cells than in the liver cells. Also, the cytotoxicity of hepatotoxicants(acetaminophen, CuCl2) was higher in the liver cells than in the kidney cells. Mitochondria potential and [Ca2+]i show compatability with cytotoxicity results in the kidney cells but not in the liver cells. The results demonstrated that nephrotoxicants can predict target organ toxicity by in vitro tests like NRU, MTT assay, mitochondria potential and [Ca2+]i. However, hepatotoxicants can predict target organ toxicity by cytotoxicity assays like NRU and MTT assays but not by mitochondria potential and [Ca2+]i.