DDS 개발과 약물의 생체내 동태 : 택솔의 세포내 거동 : in vitro 및 in vivo pharmacodynamics 에 미치는 영향

구효정 ( Kuh Hyo Jeong ),( M . Guillaume Wientjes ),( Jessie L . S . ) 한국약제학회 1997 과학의 달 기념 심포지움 Vol.- No.9

테오필린과 그 대사체의 HPLC 동시 정량 및 신(腎) 배설 특성

구효정,심창구,이민화,김신근,Kuh, Hyo-Jeong,Shim, Chang-Koo,Lee, Min-Hwa,Kim, Shin-Keun 한국약제학회 1991 Journal of Pharmaceutical Investigation Vol.21 No.1

A high-performance liquid chromatographic (HPLC) method was developed for the simultaneous determination of theophylline(TP) and its metabolites, 1-methyluric acid (1-MU) and 1,3-dimethyluric acid (1,3-DMU), in rat plasma and urine. An $100\;{\mu}l$ aliquot of a plasma or urine sample was mixed with $250\;{\mu}l$ of acetonitrite and vortexed. After centrifugation, $200\;{\mu}l$ (plasma) or $20\;{\mu}l$ (urine) aliquot of the supernatant was dried by $N_2$ stream and redissolved in $100\;{\mu}l$ (plasma) or $200\;{\mu}l$ (urine) of the mobile phase. A $20\;{\mu}l$ of the mobile phase solution was injected onto a $C_{18}$ reversed-phase column. The column was maintained at $45^{\circ}C$ by the aid of electric heating jacket. The mobile phase was a 3%(v/v) methanol solution in deionized water which contains sodium acetate (100 mM) and tetrabutyl ammonium hydroxide (4 mM). pH of the mobile phase was adjusted 4.5 by the addition of acetic acid. Detection limits for TP, 1-MU, and 1,3-DMU in plasma were 0.2, 0.1 and $0.1\;{\mu}/ml$, respectively and the corresponding values in urine were all $5\;{\mu}g/ml$. Inter- and intra-day variability of the assay for all compounds in the plasma samples was less than 5.5 and 3.8%, respectively. The retention times for 1-MU, 1,3-DMU, and TP were approximately 7, 8.5 and 18 min, respectively. Sample preparation procedure used in this method was simple, rapid and reproducible. Renal clearance of TP and its metabolites in rats showed plasma concentration dependency indicating renal tubular secretion and reabsorption of them.

혈장 시료 풀링을 통한 신약 후보물질의 흡수율 고효율 검색기법의 평가

이인경(In Kyong Yi),구효정(Hyo Jeong Kuh),정석재(Suk Jae Chung),이민화(Min Hwa Lee),심창구(Chang Koo Shim) 한국약제학회 2000 Journal of Pharmaceutical Investigation Vol.30 No.3

Genomics is providing targets faster than we can validate them and combinatorial chemistry is providing new chemical entities faster than we can screen them. Historically, the drug discovery cascade has been established as a sequential process initiated with a potency screening against a selected biological target. In this sequential process, pharmacokinetics was often regarded as a low-throughput activity. Typically, limited pharmacokinetics studies would be conducted prior to acceptance of a compound for safety evaluation and, as a result, compounds often failed to reach a clinical testing due to unfavorable pharmacokinetic characteristics. A new paradigm in drug discovery has emerged in which the entire sample collection is rapidly screened using robotized high-throughput assays at the outset of the program. Higher-throughput pharmacokinetics (HTPK) is being achieved through introduction of new techniques, including automation for sample preparation and new experimental approaches. A number of in vitro and in vivo methods are being developed for the HTPK. In vitro studies, in which many cell lines are used to screen absorption and metabolism, are generally faster than in vivo screening, and, in this sense, in vitro screening is often considered as a real HTPK. Despite the elegance of the in vitro models, however, in vivo screenings are always essential for the final confirmation. Among these in vivo methods, cassette dosing technique, is believed the methods that is applicable in the screening of pharmacokinetics of many compounds at a time. The widespread use of liquid chromatography (LC) interfaced to mass spectrometry (MS) or tandem mass spectrometry (MS/MS) allowed the feasibility of the cassette dosing technique. Another approach to increase the throughput of in vivo screening of pharmacokinetics is to reduce the number of sample analysis. Two common approaches are used for this purpose. First, samples from identical study designs but that contain different drug candidate can be pooled to produce single set of samples, thus, reducing sample to be analyzed. Second, for a single test compound, serial plasma samples can be pooled to produce a single composite sample for analysis. In this review, we validated the issue whether the second method can be applied to practical screening of in vivo pharmacokinetics using data from seven of our previous bioequivalence studies. For a given drug, equally spaced serial plasma samples were pooled to achieve a Pooled Concentration for the drug. An area under the plasma drug concentration-time curve (AUC) was then calculated theoretically using the pooled concentration and the predicted AUC value was statistically compared with the traditionally calculated AUC value. The comparison revealed that the sample pooling method generated reasonably accurate AUC values when compared with those obtained by the traditional approach. It is especially noteworthy that the accuracy was obtained by the analysis of only one sample instead of analyses of a number of samples that necessitates a significant man-power and time. Thus, we propose the sample pooling method as an alternative to in vivo pharmacokinetic approach in the selection potential lead(s) from combinatorial libraries.

인체폐암세포 조직배양계(histocultures)에서 티라파자민의 약력학

박종국 ( Jong Kook Park ),구효정 ( Hyo Jeong Kuh ) 한국약제학회 2006 Journal of Pharmaceutical Investigation Vol.36 No.4

인체대장암 세포에서 후성적 유전자 불활성화 저해제와 5-Fluorouracil의 병용효과분석

김미영(Mi Young Kim),손정규(Jung Kyu Son),이숙경(Suk Kyung Lee),구효정(Hyo Jeong Kuh) 大韓藥學會 2005 약학회지 Vol.49 No.6

Low sensitivity to anticancer drugs such as 5-fluorouracil (5-FU) has been associated with decreased expression of genes involved in cell proliferation, apoptosis and metastasis. Recently, it has been shown that the expression levels of some of these genes are reduced by transcription inhibition due to epigenetic silencing on CpG islands. Therefore, epigenetic therapy has been proposed, where epigenetic silencing is repressed with DNA methyltransferase (DNMT) inhibitors and histone deacetylase (HDAC) inhibitors alone or in combination with other chemotherapeutic agents. The aim of our study was to evaluate the combination effect of 5-FU and its association with the status of epigenetic silencing using methylation-specific PCR of p14ARF when given with 5-aza-2'-deoxycytidine (5-aza-dC), a DNMT inhibitor and depsipeptide, and HDAC inhibitor in DLD-1 human colorectal cancer cells. The combination of 5-aza-dC with depsipeptide showed a synergism and induced unmethylation of p14ARF. However, triplet combination of 5-aza-dC/epsipeptide and 5-FU resulted in antagonistic effects and abrogated unmethylation of p14ARF. These results suggest that unfavorable interaction of 5-aza-dC/depsipeptide with 5-FU in DLD-1 cells may be related with the failure in repression of epigenetic silencing, which warrants further investigation.

인체 대장암세포 다층배양계에서 파크리탁셀의 투과

최미선 ( Mi Sun Choi ),박종국 ( Jong Kook Park ),알압드아메드 ( Ahmed M. AL Abd ),구효정 ( Hyo Jeong Kuh ) 한국약제학회 2006 Journal of Pharmaceutical Investigation Vol.36 No.6

혈액 투석을 받는 두경부암 환자의 동시화학방사선요법에서 Cisplatin의 약력학 조사 1예

김수지(Suzy Kim),전연주(Younjoo Jeon),이호상(Ho-Sang Lee),이상훈(Sang-Hun Lee),김형욱(Hyung Wook Kim),심병용(ByoungYong Shim),구효정(Hyo-Jeong Kuh),김훈교(Hoon-Kyo Kim),박철휘(heol Whee Park) 대한두경부종양학회 2007 대한두경부 종양학회지 Vol.23 No.2

목 적 :투석을 받고 있는 진행된 두경부암 환자에서 Cisplatin동시 화학방사선 요법의 타당성과 약물역동학에 대해 연구 하였다. 방 법 :57세 말기 신장병을 앓고 있던 남자환자가 임상 병기 3기의 외이도 암을 진단 받고 종양 완전 절제술을 시행하였다. 수술 후 6개월에 환자는 국소 부위 재발하였다. 절단술과 술후 5200cGy, 26fx의 방사선 치료 시행에도 불구하고 다시 국소 부위 재발 보여 3주기동안 Cisplatin 기반의 방사선 화학 병합 요법 치료를 하였다. 총 3주간 Cisplatin 20 mg/m 2 를 투여했으며 화학 요법 1일째 30분 동안 Cispla-tin을 정주하고 30분 후 혈액투석을 4시간 동안 시행하였다. 화학요법 3일째와 5일째에도 혈액투석을 시행하였다. 환자의 혈장 시료는 Cisplatin 정주 후 특정한 시간에 채취되었다. 결 과 :Cisplatin 동시 화학방사선요법 3차 시행 후 환자의 종 괴 크기는 현저하게 감소하였다. Cisplatin의 최대 혈장 농도는 총 platinum은 0.74μg/ml였고 free platinum은 0.37μg/ml로 이었다. AUC 값은 총 platinum은 94.7μg·h/ml, free platinum은 11.3 μg·h/ml이었다. 결 론 :투석을 받는 진행된 두경부암 환자에서 Cisplatin 동시 화학방사선요법을 시행한 증례를 보고하며 이들 환자에서 정용량의 Cisplatin 동시 화학방사선요법이 감내할 만하다고 제안한다. Objectives :We study the feasibility and pharmacokinetics of cisplatin concurrent chemoradiation for ad-vanced head and neck cancer patient undergoing hemodialysis. Materials and Methods :A 57-year old male with end stage renal disease developed stage III external auditory canal cancer. Complete resection surgery was done. Postoperative 6 months, local recurrence was occurred. De-spite excision and adjuvant radiotherapy, local tumor was recurred. We decided to treat a cisplatin concurrent chemoradiotherapy. Cisplatin was administered at a dose of 20mg/m 2 for 30 min. Hemodialysis was started 30 min after completion of the cisplatin infusion and performed for 4 hours. Hemodialysis was performed on day 3 and 5 of chemotherapy. Plasma samples were collected at specified times after administration of cisplatin. Result :At the end of the third cycle of cisplatin concurrent chemoradiotherapy, the tumor size was mark-edly decreased. The maximum plasma concentrations of plasma platinum and free platinum were 0.74 and 0.37μg/ml respectively. The area under the curve of plasma platinum and free platinum were 94.7 and 11.3μg·h/ml respectively. Conclusion :We report a case of Cisplatin concurrent chemoradiation for hemodialysis patient with advanced head and neck cancer and suggest full dose cisplatin concurrent chemoradiotherpay is tolerable for these patients

Theopyylline 및 대사체의 HPLC 동시 정량 및 Rat에 있어서의 신배설 기전에 관한 연구

구효정 보건장학회 1989 보건장학회보연구논문집 Vol.11 No.-

셀레콕시브 및 그 합성유도체들의 항암활성 스크리닝

박정란,강진형,구효정,노지영,류형철,박상욱,고동현,조일환,이주영,황다니엘,김인경 한국약제학회 2003 Journal of Pharmaceutical Investigation Vol.33 No.2

Selective COX (cyclooxygenase)-2 inhibitors including celecoxib have been shown to induce apoptosis and cell cycle changes in various tumor cells. New inhibitors are recently being developed as chemomodulating agents. We evaluated celecoxib and screened 150 synthetic compounds for anti-proliferative activities in vitro. Effects of celecoxib on COX activity, cell growth, cell cycle distribution, and apoptosis induction were determined in A549 COX-2 overexpressing human non-small cell lung cancer (NSCLC) cells. The COX inhibition of celecoxib increased with concentration up to 82% at 1μM after 24 hr exposure. Forty μM and 50μM of celecoxib induce G_1 arrest, and TUNEL-positive apoptotic cells, respectively. Among 150 compounds, several compounds were selected for having greater COX-2 inhibitory activity and higher selectivity than celecoxib with growth inhibitory activity. Celecoxib showed concentration-dependent COX inhibitory activity, and ability to induce cell cycle arrest and apoptosis in human NSCLC cells in vitro. Among synthetic analogues screened, several compounds showed promising in vitro activity as COX-2 inhibitory anticancer agents, which warrant further evaluation in vitro and in vivo.

항고형암제의 활성평가를 위한 in vitro 삼차원 암세포 배양계의 확립

이상학,이주호,구효정 한국약제학회 2004 Journal of Pharmaceutical Investigation Vol.34 No.5

For the efficient determination of activity against solid tumors, an in vitro tumor model that resembles the condition of in vivo solid tumors, is required. The purpose of this study was to establish a rapid culture method and viability assay for an in vitro 3-dimensional tumor model, multicellular spheroid (MCS). Among 12 human cancer cell lines, a few cell lines including DUD-1 (human colorectal carcinoma cells) formed fully compact MCS which was adequate for in vitro viability assay. DLD-1 MCS showed steady growth reaching 700 pm diameter after 11 day culture. DUD-1 cells grown as MCS showed significant increase in G_(0)/G₁ phase compared to the monolayer cells (73.9% vs 45.7%). but necrotic regions or apoptotic cells were not observed. The cells cultured as MCS showed resistance to 5-FU (10.3 fold higher IC_(50)) compared to monolayers. however, tirapazamine (a hypotoxin) showed similar activity in both culture systems. In summary, MCS may be a valid in vitro model for activity screening of anticancer agents against human solid tumors and also exploitable for studying molecular markers of drug resistance in human solid tumors.