PC12 세포에서 신경전달물질 방출을 저해하는 저분자 생리활성물질 S8877의 탐색

강주섭,지옥화,이윤식 대한암예방학회 2006 Journal of cancer prevention Vol.11 No.1

We established an in vitro experimental system using the following procedure. We first introduced tritium-labeled norepinephrine ([3H]-NE) into PC12 cells. The [3H]-NE incorporated into PC12 cells were then stimulated by a high concentration (60 mM) of K+ buffer during 12 minutes. Then, we collected 100μl supernatant and counted the amount of [3H]-NE release from PC12 cells with a scintillation counter. After screening fungal, Streptomyces spp. or bacterial product using this experimental system, we obtained S8877 from Streptomyces spp. which inhibited [3H]-NE release from PC12 cells. S8877 also inhibits the release of ATP as a neurotransmitter of PC12 cells and rat cortical neurons. The inhibitory effect was seen even when the PC12 cells were treated with low K+ buffer containing ionomycin (1μM) as an ionophore. This result suggests that the inhibitory action of S8877 on neurotransmitter release appeared after the influx of Ca2+. (Cancer Prev Res 11, 22-29, 2006)

Curcumin, indomethacin, and naproxen synergistically inhibit the growth of human colorectal cancer cells (HT-29)

강주섭,Hyun-Jin Kim,오현인,Su-Jin Lim,Jung-Wook Hong,Jin-Hee Park,Hai-Dan Yuan 대한암예방학회 2007 Journal of cancer prevention Vol.12 No.1

The aim of this study was to investigate the effect of curcumin, indomethacin, curcumin and naproxen combination synergistically on the growth of HT-29 cells. The growth of HT-29 cells were examined by MTS cell viability assay by one to four days treatment with curcumin (10~50 uM), indomethacin (250~1,000 uM), curcumin (10~50 uM) and naproxen (250~1,000 uM) to 2×104 HT-29cells/mL. The results suggested that curcumin inhibited the growth of HT-29 cells in a dose-dependent fashion. In concentration range from 500 to 1,000 uM, indomethacin was inhibitied the growth of HT-29 cells in a dose - dependent fashion. To examine inhibitory effects of curcumin and indomethacin, curcumin and naproxen combinination on the HT-29 cell lines, we treatment with 20 uM curcumin and indomethacin (500 uM, 750 uM) combinination curcumin and naproxen (750 uM, 1,000 uM) combinination to 2×104 HT-29 cells/well. The results suggested that curcumin potentiates the inhibitory effect of indomethacin on the growth of HT-29 cell lines by shifting the dose-dependent manner. (Cancer Prev Res 12, 30-34, 2007)

High-Throughput Analytic Method Development for Gabapentin in Human Plasma by LC Coupled with Tandem Mass Spectrometry for Pharmacokinetic Studies

강주섭,Hee-Kyung Seo,Song-Hee Park,Jung-Sik Lee,Su-Jin Lim,Jin-Hee Park,Min-Ho Lee,Ok-Hwa Jhee 대한암예방학회 2007 Journal of cancer prevention Vol.12 No.1

A simple, sensitive and validated liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for gabapentin (GBP) in human plasma has been developed for pharmacokinetic (PK) studies. In a single-dose PK study of GBP in normal 24 subjects were given single oral doses of 300 mg GBP after fasting 12 hours. Plasma samples were drawn predose and then serially at 13 times after dose for 36 hours and plasma GBP were analyzed by deveopled LC-MS/MS. The procedures involves a simple protein precipitation with acetonitrile (ACN) and separated by LC equipped with C18 column using ACN: 10 mM ammonium acetate (20:80, v/v, pH adjusted to 3.2 with acetic acid) as mobile phase. The GBP and α-aminocyclohexane propionic acid hydrate (IS, internal standard) were analyzed by API 2000 MS/MS in the multiple-reaction monitoring (MRM) mode. The MS/MS analysis was optimized using ESI (+) and selectivity with ionization as m/z 154.0 for GBP and m/z 126.0 for IS that fully separated during a 4.0-min run time. The accuracy and precision were met the analytical method validation criteria for concentrations over the standard ranges. The calibration curve was linear over a working range of 20(LLOQ) - 5,000 ng/ml and no interfering endogenous compounds with the analysis. This LC-MS/MS method was successfully applied for the GBP PK studies after single oral dosing of 300 mg of GBP in 24 Korean healthy subjects. (Cancer Prev Res 12, 41-50, 2007)

Effect of Korean Oriental Medicine Extract on Bone Mass as Compared with Alendronate in Ovariectomized Rats

강주섭,이주원,지옥화,Haidan Yuan,김태화,김동선,이민호,Ae-Son Om,이보배,박성국 한국식품영양과학회 2005 Journal of medicinal food Vol.8 No.3

A number of alternative medicines (AMs) have often been used as traditional therapies for various diseaseswithout scientific or clinical evidence supporting their use. The present study examined the pharmaceutical effects of an AMextract with a long history of use as a traditional medicine for various bone diseases. To evaluate it as a potential candidatefor use as an anti-osteoporotic drug, we investigated the effects of the AM extract on the progression of bone loss in ovariec-tomized (OVX) rats fed a calcium (Ca)-deficient diet for 4 or 12 weeks. We also compared the AM extract with alendronate,an anti-resorptive drug. The AM extract did not influence bone turnover as indicated by biochemical markers [i.e., de-oxypyridinoline (DPD)]. In contrast, alendronate treatment seemed to reduce bone turnover via inhibition of bone resorptionas evidenced by reduced urinary DPD concentrations accompanied by a tendency for decreased serum tartrate-resistant acidphosphatase. Administration of alendronate or AM extracts did not significantly affect bone density, although both tended toincrease bone mineral density (BMD) and bone strength of the femur. Although both treatments did not affect vertebral BMDand bone strength, histological analysis of vertebrae showed well-developed trabecular networking in OVX rats treated withalendronate or AM extract, in contrast to the thin and disconnected trabecule in OVX rats. In conclusion, the AM extract pro-duced a very weak effect on the prevention of bone loss induced by OVX and Ca deficiency in rats, but was similar to theresults observed with alendronate. Further verification is necessary to justify the use of the AM extract as a treatment for os-teoporosis.

약물대사효소계로서의 Flavin-Containing Monooxygenases(FMOs)

강주섭,이민호 한양대학교 의과대학 2001 한양의대 학술지 Vol.21 No.1

Together with the cytochrome P-450(CYPs) containing mixed function oxidases, the flavin-containing monooxygenase(FMOs) included in mammalian tissue microsomes play a major role in the initial oxidation of xenobiotics containing soft-nucleophiles, including alkaloids, pesticides and drugs to polar metabolite for easier conjugation and excretion. These include tertiary and secondary alkyl- and arylamines, many hydrazines, thiocarbamides, thioamides, sulfieds, disulfides, thiols and other soft nucleophiles. This flavinprotein that catalyzes NADPH- and oxygen-dependent oxidation of a wide range of xenobiotics and durgs is typical for the enzyces of detoxification. The FMO share a mechanism distinctly different from other oxidases bearing flavin, heme or other redox-active prosthetic groups. Several major advances has been made during the last 10 years regarding our understanding of the FMOs, including their catalytic properties, role inmetabolic transformation of xenobiotics to both less and more toxic metabolites, molecular structure, expression patterns, and phenotypes of its activity. There is no question that FMOs are fundamentally different from the enzymes of biosynthesis in that precise binding, tight fit of the xenobiotic substrate to the enzyme is not necessary for catalysis. Therefore, We try to illustrate the various aspect of FMOs characteristics such as substrate specificity, stereoselectivity, molecular characterization and regulation of expression, bioactivation of xenobiotics, contemporary issues in drug metabolism in human in this review.

Streptozotocin으로 당뇨 유발 신생 흰쥐에서 Leucocyanidins의 투여가 혈당 및 혈액학적 지수에 미치는 영향

강주섭,박진희,바규신,김신희,지옥화,이정식,박송희,서희경,임수진 대한암예방학회 2007 Journal of cancer prevention Vol.12 No.1

To investigate the effect of leucocyanidins (LCN) in streptozotocin (STZ)-treated newborn diabetic rats, we evaluated the changes of body weight, fasting blood glucose level, oral glucose tolerance test (OGTT), insulin level and biochemical parameters after the administration of LCN in STZ-treated newborn rats. In 7 days of age rats were received single intraperitoneal injection of 60 mg/kg of STZ. Animals were divided into four groups; STZ-induced DM group; LCN (LL)-treated group (30 mg/kg, i.p), LCN (HL)-treated group (60 mg/kg, i.p) at 8 weeks after the single injection of STZ (60 mg/kg, i.p.), respectively during 4 weeks.; Normal control (NC) group received vehicle and saline. Mean body weight change of DM group was retarded greatly by STZ-exposure. While, body weights of LL and HL groups were increased. Fasting blood glucose levels of LL and HL groups were reduced about 19% or 27% by LCN injection. The insulin level did not differ after treatment with LCN in experimental rats. The results of OGTT was improved in HL group compared to DM group. In LL and HL groups, blood glucose, ALT and AST and BUN levels were attenuated compared to DM group. From the results, it was suggested that LCN had a tendency to decrease on STZ-induced toxicity in terms of monitoring the blood glucose in fasting, OGTT and some biochemical parameter of the rat. (Cancer Prev Res 12, 51-57, 2007) Key Words: Streptozotocin, Leucocyanidins, Blood glucose, OGTT, Diabetes

흰쥐에서의 Dextran Sulfate Sodium-유발 궤양성 대장염에 대한 Curcumin의 효과

강주섭,원해단,지옥화,김신희,김현진,엄애선,이윤식,백승삼,김종욱 대한암예방학회 2006 Journal of cancer prevention Vol.11 No.2

Ulcerative colitis (UC), one of the inflammatory bowel diseases (IBD), tends to increase in Korea every year, but their causes and pathogenesis is still unknown. Sulfasalazine and corticosteroid are regularly used to treat IBD, but they have many side effects during long-term use and none therapeutic effect for some patients. So it is important to explore new agents that are more effective and have fewer side effects. Curcuma is the main component of turmeric, has anti-carcinogenic, anti-oxidative and anti- inflammatory activities. The aim of this study was to evaluate the effect of curcumin on UC induced by dextran sulfate sodium (2% DSS for 4 weeks) in the rat. The experiment animals were divided into four groups: control (normal), DDS-induced colitis rats, high-dose (100 mg/kg/day, po) and low dose (10 mg/kg/day, po)-treated groups that treated for 7 weeks during 4 weeks with 2% DDS and 3 weeks alone. We evaluated pathologically disease activity index (DAI) and the colon mucosa damage index (CMDI) and biochemically malondialdehyde (MDA), prostaglandin E2 (PGE2) in colon mucosa. The DAI and CDMI in DDS-induced colitis rats were more severe and the content of MDA and PGE2 level in DDS-induced rat were also higher than control and curcumin-treated rats, respectively. Curcumin may be alleviating some abnormality of disease index in a dose-depended manner. (Cancer Prevention Res 11, 137-143, 2006)

DMN-유발 간섬유화 동물에서 혈류 의존성 약물인 Verapamil의 약물동태에 대한 간질환의 영향

강주섭,박윤영,원해단,김현진,이주원,지옥화,엄애선,이상구,이윤식,신인철,이민호,홍정욱 대한암예방학회 2005 Journal of cancer prevention Vol.10 No.1

The purpose of this study was to determine the effect of hepatic disease severities on pharmacokinetics of flow-limited verapamil in dimethylnitrosamine (DMN)-induced cirrhotic rats. Hepatic cirrhotic rats (SD rats, 200~250 g) were induced by intraperitoneal injection with 1% DMN solution at a dose of 10μg/g by 3 days/week as experimental groups and same dose of saline as control for 4 weeks. The serum verapamil concentrations were quantified at zero, 10 min, 30 min, 1, 2 and 3 hrs after bolus injection of 3 mg/kg of verapamil by a column-switching HPLC method and pharmacokinetic parameters such as Co, MRT, AUC, Vdss, t1/2 (β) and CLp were determined in each group. And then some hepatic tissue was obtained and subjected to analysis of the hepatic 4-hydroxyproline content and were inspected by light microscope after hematoxylin and eosin staining. The serum concentrations and pharmacokinetic parameters such as Co, AUC, MRT and t1/2 (β) were significantly increased (p<0.01) and Vdss and CLp were significantly decreased (p<0.01) in hepatic cirrhotic rats according to DMN-treated period. The 4-hydroxyproline content was also gradually increased propertionally to DMN-treated period. The pharmacokinetic parameters of verapamil were seemed to change gradually to depend on the hepatic fibrotic severity. Therefore, we suggested that flow-limited drug as like verapamil dosage adjustment is necessary according to hepatic function severity. (Cancer Prev Res 10, 34-47, 2005)

신경전달물질 방출 저해제 S8877이 신경세포와 PC12 세포의 돌기신장에 미치는 영향

강주섭,김종욱,지옥화,엄애선,이윤식 대한암예방학회 2006 Journal of cancer prevention Vol.11 No.1

We identified S8877, a novel microbial metabolite from Streptomyces spp., to inhibit the release of neurotransmitter form rat hippocampal neuron and PC12 cells. S8877 is an inhibitor of trifiated norepinephrine ([3H]-NE) release in high K+ buffer solution containing ionomycin, indicating that S8877 inhibits neurotransmitter release after the influx of Ca2+ ions. We also examined the effect of S8877 on β-glucuronidase release from guinea pig neurophils and on the neurite extension of rat hippocampal neurons and PC12 cells. As a result, S8877 increased β-glucruonidas release: when treated with 5μg/ml of S8877, which prevented [3H]-NE release, and the inhibition of neurite extension for both rat hippocampal neurons and PC12 cells was observed. (Cancer Prev Res 11, 30-38, 2006)

Modern Methods for Analysis of Antiepileptic Drugs in the Biological Fluids for Pharmacokinetics, Bioequivalence and Therapeutic Drug Monitoring

강주섭,Yoo-Sin Park,Shin-Hee Kim,Sang-Hyun Kim,전민영 대한약리학회 2011 The Korean Journal of Physiology & Pharmacology Vol.15 No.2

Epilepsy is a chronic disease occurring in approximately 1.0% of the world’s population. About 30% of the epileptic patients treated with availably antiepileptic drugs (AEDs) continue to have seizures and are considered therapy-resistant or refractory patients. The ultimate goal for the use of AEDs is complete cessation of seizures without side effects. Because of a narrow therapeutic index of AEDs, a complete understanding of its clinical pharmacokinetics is essential for understanding of the pharmacodynamics of these drugs. These drug concentrations in biological fluids serve as surrogate markers and can be used to guide or target drug dosing. Because early studies demonstrated clinical and/or electroencephalographic correlations with serum concentrations of several AEDs, It has been almost 50 years since clinicians started using plasma concentrations of AEDs to optimize pharmacotherapy in patients with epilepsy. Therefore, validated analytical method for concentrations of AEDs in biological fluids is a necessity in order to explore pharmacokinetics, bioequivalence and TDM in various clinical situations. There are hundreds of published articles on the analysis of specific AEDs by a wide variety of analytical methods in biological samples have appears over the past decade. This review intends to provide an updated, concise overview on the modern method development for monitoring AEDs for pharmacokinetic studies, bioequivalence and therapeutic drug monitoring.