인체 간 microsome에서 pentoxifylline 대사체 M-1의 시험관내 대사

신혜순(Hea Soon Shin) 대한약학회 1999 약학회지 Vol.43 No.6

The metabolism and pharmacokinetics of M-1, which is metabolite of pentoxifylline, have been studied in human liver microsomes. Biphasic kinetics was observed from the Eadie-Hofstee plot for the formation of both metabolites of M-1. For the kinetics of pentoxifylline, mean values of Vmax1 and Vmax2 were 1.648 and 5.622nmol/min/mg protein, and the estimated values of Km1 and Km2 were 0.180 and 4.829mM, respectively. For M-3, mean values of Vmax1 and Vmax2 were 0.062 and 0.491nmol/min/mg protein, and estimated values of Km1 and Km2 were 0.025 and 1.216mM. The formations of pentoxifylline and M-3 from M-1 were identified by using several selective inhibitors of cytochrome P450 isoformes at 0.05-5mM concentratibn of M-1 in human liver microsomes. For the analysis of low (0.05mM) concentration of M-1, where the affinity was expected as low, indicated that CYP1A2 and CYP3A4 were major P450 isoforms responsible for pentoxifylline and M-3 formation. CYP3A4 and CYP2A6 appeared to be P450 isoforms responsible for M-3 formation at high (5mM) concentration of M-1.

퇴행성 뇌질환 치료제 Tacrine 유도체의 프로스타글란딘 생합성 억제효과

신혜순(Hea Soon Shin) 대한약학회 2005 약학회지 Vol.49 No.1

Tacrine analogues for degenerative brain disease treatments have been designed. A series of diazaanthrine derivatives as novel analogues of tacrine has been prepared through the alkyl substitution and the ring expansion. They were expected to retain anti-inflammatory activity by inhibition of prostaglandin production with reduction of side effect as the selective prostaglandin synthase inhibitor. Prostaglandin synthase expression is associated with the deposition of beta-amyloid protein in neuritic plaques in brain inflammation. Therefore selective prostaglandin synthase blockade is important for the prevention and treatment of alzheimer's disease. To evaluate inhibitory effect of prostaglandin synthase, synthetic tacrine derivatives were screened with accumulation of prostaglandin biosynthesis by lipopolysaccharide in aspirin-treated murine macrophage cell. Most of synthetic compounds have shown significant prostaglandin synthase activities in vitro screening with 84.3~33.6% inhibition of the prostaglandin E₂ production at 10 ㎍/ml.

Benzothiazine-3-carboxamide 유도체의 합성과 CGX-2 저해효과

신혜순(Hea Soon Shin),최희전(Hee Jeon Choi),권순경(Soon Kyoung Kwon) 대한약학회 2002 약학회지 Vol.46 No.6

In tai s study, newly de signed COX-2 inhibitors , synthetic derivatives of benzothiazine-3-carboxamide, were screened in vitro for selectivity of COX-1 and COX-2 inhibition properties. 7-Bromo-1,2-benzoisothiazine derivatives were obtained from 4-bromotoluene over the chlorosulfonation, amination and oxidation. And benzothiazine ring was synthesized through Gabriel-Colmann rearrangement reaction. To evaluate inhibitory erect of COX-2, synthetic derivatives of benzothiazine-3- carboxamide were tested with accumulation of prostaglandin by lipopolysaccharide in aspirin-treated murine macropharge cell. Some of the synthesized lead compounds have potentially shown the structure-activity relationship for selectivity of COX-2 inhibition activity.

Pyrimidine Nucleoside 유도체들의 합성 및 약물학적 효능 검색

신혜순(Hea Soon Shin),이희주(Hee Joo Lee) 한국응용약물학회 1995 Biomolecules & Therapeutics(구 응용약물학회지) Vol.3 No.3

Several acyclonucleoside analogues of pyrimidine base and N¹-derivatives of 5-fluorouracil have been synthesized and evaluated for their biological effects. When tested with in vitro Lekemia L1210 cells, the 5-fluorouracil derivatives exhibited slightly higher antitumor activity than the parent 5-fluorouracil. When tested against Herpes Simplex Virus type 1 and type 2 cultured in the Vero cell, the 5-fluorouracil derivatives showed weak antiviral activity.

Cyclooxygenase-2 저해제로서의 benzothiazine 유도체 합성과 항염작용 평가

신혜순(Hea Soon Shin),박명숙(Myung Sook Park),권순경(Soon Kyoung Kwon) 대한약학회 2000 약학회지 Vol.44 No.3

The antiinflammatory mechanism of NSAIDs is attributed to the reduction of prostaglandin synthesis by the direct inhibition of cyclooxygenase. Inhibition of prostaglandin production in organs such as stomach and kidney can result in gastric lesions, nephrotoxicity and increased bleeding. In this study, newly designed COX-2 inhibitors, synthesized 1,2-benzothiazine derivatives, were screened in vitro for selectivity of COX-1 and COX-2 inhibition properties. Lead compounds in the structure-activity relationship were studied to synthesize new highly selective COX-2 inhibitors. To determine inhibitory effect of COX-2, synthesized 1,2-benzothiazine derivatives were screened with accumulation of prostaglandin by lipopolysaccharide (LPS) in aspirin-treated macrophages and murine macropharge cell. Some of synthesized 1,2-benzothiazine derivatives were shown to be effective as selective COX-2 inhibitory activity. Others exhibited a preferential inhibition of COX-2, although some COX-1 inhibitory activity was still present. As a conclusion, simple monomer derivatives were more active than dimer derivatives. Substitution of halogen (Br, Cl) on the benzothiazine nucleus slightly enhanced inhibition activity.

Glucuronidation of Hydroxylated Polychlorinated Biphenyl by Channel Catfish Liver

Hea-Soon Shin(신혜순) 환경독성보건학회 2008 환경독성보건학회지 Vol.23 No.3

Hydroxylated polychlorinated biphenyl (OH-PCBs)는 PCB의 CYP-dependent oxidation의 대사물로서 잠재적 독성이 강하고 지질친화성을 가지며 생물군에 지속적인 축적성을 나타낸다. OH-PCBs의 해독화 효능을 조사하기 위하여 channel catfish 간에서 glucuronidation을 통한 해독작용 가능성을 평가하고 biphenyl 구조에 다양한 염소치환의 구조적 차이점에 따른 영향을 비교 분석하여 보았다. Kinetic parameters에서 Km과 V<SUB>max</SUB>는 192~871 μM, 869~1774 pmol/min/mg으로써 4′-OH-PCB35와 4′-OHPCB69이 가장 높은 속도의 glucuronidation을 나타냈으며, 구조적 차이점에서 phenolic group에 한 개의 염소치환이 존재할 경우보다 두 개의 염소치환이 존재할 경우에 OH-PCBs (p<0.001)의 glucuronidation에 대한 V<SUB>max</SUB>를 현저하게 낮추는 결과를 보였다.

Probiotics with Antimicrobial Activity against Multidrug Resistant Pseudomonas aeruginosa and Acinetobacter baumannii

이도경,김민지,강주연,박재은,신혜순,하남주,Lee, Do Kyung,Kim, Min Ji,Kang, Joo Yeon,Park, Jae Eun,Shin, Hea Soon,Ha, Nam Joo The Microbiological Society of Korea 2013 미생물학회지 Vol.49 No.3

녹농균과 아시네토박터 바우마니는 병원에서 중요한 기회감염 균주이며 대부분의 항생제에 내성이다. 다제내성 녹농균(MDRPA)과 아시네토박터 바우마니(MDRAB)는 심각한 원내 감염을 일으키고 메티실린 내성 황색포도상구균(MRSA) 보다 치료가 어렵다. 비피도박테리아는 많은 유익한 프로바이틱스 중 하나로 그들의 항균활성에 대한 많은 연구가 이루어져 왔다. 본 연구에서는 한국인으로부터 분리한 비피도박테리움 속 균주들의 MDRPA와 MDRAB에 대한 항균활성을 조사하였다. 시프로플록사신, 토브라마이신, 겐타마이신, 메로페넴과 세프타지딤에 내성을 보이는 MDRPA와 MDRAB에 대한 비피도박테리움 속 균주들의 항균활성은 흡광도를 이용한 액체배지희석법에 의해 측정되었다. 모든 비피도박테리아 분리균주(비피도박테리움 어돌레센티스 9균주, 비피도박테리움 롱검 3균주와 비피도박테리움 슈도카테눌라툼 2균주) 중 비피도박테리움 슈도카테눌라툼 SPM1309의 배양 상등액은 MDRPA와 MDRAB에 대한 강한 증식 억제 효과를 보여주었다. 배양 시간 동안 혼합액의 탁도는 변화되지 않았으며, 이 억제 효과는 정균 작용이었다. 게다가 항균활성은 분자량 10 kDa 미만의 분획물에서 나타났으며, $70^{\circ}C$에서 10분간 열처리 한 후에서 항균활성이 유지되었기 때문에 활성물질은 열에 안정하였다. 본 연구결과는 MDRPA와 MDRAB 감염증의 대체치료법을 위한 비피도박테리움 슈토카테눌라툼 SPM1309의 잠재적인 가능성을 보여준다. Pseudomonas aeruginosa and Acinetobacter baumannii are significant opportunistic pathogens in hospitals and are resistant to most antibiotics. Multidrug-resistant P. aeruginosa (MDRPA) and A. baumannii (MDRAB) cause severe human nosocomial infections and are more difficult to treat than methicillin-resistant Staphylococcus aureus (MRSA). Bifidobacteria are among of the most beneficial probiotics and have been widely studied for their antimicrobial activities. The present study explored the antimicrobial activity of Bifidobacterium sp. isolated from healthy Koreans against MDRPA and MDRAB. The antimicrobial activity of the isolates against MDRPA and MDRAB, which are resistant to ciprofloxacin, tobramycin, gentamicin, meropenem, and ceftazidime, was determined by modified broth microdilution methods using absorbance. Among all tested bifidobacteria isolates (nine B. adolescentis, three B. longum, and two B. pseudocatenulatum), the culture supernatant of B. pseudocatenulatum SPM1309 showed a strong growth inhibitory effect against MDRPA and MDRAB. No change in the turbidity of the mixture was observed during incubation, and its inhibitory effect occurred through bacteriostastic action. Moreover, the antibacterial activity was observed in the fraction with molecular weights <10 kDa of bifidobacteria culture supernatant, and the active fraction was heat-stable because it maintained its activity when heated at $70^{\circ}C$ for 10 min. The results suggest that this Bifidobacterium strain could have potential applications for alternative therapy in MDRPA and MDRAB infections.

2010년도 국내 임상에서 분리한 다제내성 녹농균의 유전자형 조사

김민지,차민경,이도경,강주연,박재은,김영희,박일호,신혜순,하남주,Kim, Min Ji,Cha, Min Kyeong,Lee, Do Kyung,Kang, Ju Yeon,Park, Jae Eun,Kim, Young Hee,Park, Il Ho,Shin, Hea Soon,Ha, Nam Joo The Microbiological Society of Korea 2012 미생물학회지 Vol.48 No.4

녹농균은 특히 면역이 저하된 환자에게서 심각한 감염을 일으키는 그람음성의 기회감염 균주이다. 또한 carbapenem 내성 metallo-${\beta}$-lactamases (MBL)를 가진 녹농균이 한국에서 증가되는 추세로 보고되고 있다. 따라서 본 실험에서는 2차 병원인 삼육 서울 병원에서 수집된 총 92종의 임상 녹농균의 다재내성 수준을 분석하였다. 항생제에 대한 감수성은 최소억제농도(MIC) 분석에 의해 결정되었고, inhibitor-potentiated disk diffusion(IPD) 분석은 MBL 검출을 위해 수행되었다. RAPD-PCR은 임상환자에서 분리한 녹농균 계통의 유전적 유형의 특징을 밝히기 위해 사용되었다. 그 결과 임상에서 분리된 녹농균의 40.2%는 ceftazidime에 내성을, 58.7%는 meropenem에 내성을, 56.5%는 gentamicin에 내성을, 46.7%는 tobramycin에 내성을, 62.0%는 ciprofloxacin에 내성을 그리고 97.8%는 chloramphenicol에 내성을 보였다. IPD 분석에 의해 29종의 다재내성 균주로 관찰 되었고, RAPD 분석에 의해 19종은 IPM-1 유전자형을, 2종은 VIM-2 유전자형을 만들었다. MBL 유전자 검출 시험을 통해 19종의 IMP-1 생성 녹농균 중에서 16종이 유사한 유전자형을 보였고, 3종은 다른 유전자형이 관찰되었다. 임상에서 분리한 IMP-1 생성 다재내성 녹농균의 비율은 꾸준히 증가하고 있다. 이번 연구는 2010년 국내 임상에서 분리한 녹농균의 항생제 다제내성 패턴과 유전자형에 대한 정보를 제공한다. Pseudomonas aeruginosa is an opportunistic Gram-negative bacterium that causes serious infection, particularly in immunocompromised patients. Also, P. aeruginosa possessing carbapenem-resistant metallo-${\beta}$-lactamases (MBL) has been reported with increasing frequency in Korea. We therefore analyzed the level of multidrug-resistant clinical P. aeruginosa isolated from a secondary hospital in Korea in 2010. A total of 92 isolates of P. aeruginosa were collected from Sahmyook Medical Center in 2010. Susceptibility to antimicrobial agents was determined by analysis of the minimum inhibitory concentration test; the inhibitor-potentiated disk diffusion (IPD) test was performed for MBL detection. RAPD-PCR was used for genotyping to rapidly characterize P. aeruginosa strains isolated from clinical patients. The percentages of non-susceptible isolates were as follows: 40.2% to ceftazidime, 58.7% to meropenem, 56.5% to gentamicin, 46.7% to tobramycin, 62.0% to ciprofloxacin and 97.8% to chloramphenicol. The 29 multidrug-resistant strains were screened by the IPD test: of the 21 PCR-positive isolates, 19 were IPM-1 producers and 2 were VIM-2 producers. Among the 19 IMP-1-producing P. aeruginosa isolates, 16 isolates showed similar patterns, and three different banding patterns were observed. The proportion of IMP-1-producing multidrug-resistant P. aeruginosa from clinical isolates steadily increased in this secondary hospital in Korea in 2010. This study provides information about the antimicrobial-resistant patterns and genotype of multidrug-resistant P. aeruginosa isolated from clinical isolates in Korea, 2010.

Dual Coating Improves the Survival of Probiotic Bifidobacterium Strains during Exposure to Simulated Gastro-Intestinal Conditions

강주연,이도경,박재은,김민지,이중수,서재구,정명준,신혜순,하남주,Kang, Joo Yeon,Lee, Do Kyung,Park, Jae Eun,Kim, Min Ji,Lee, Joong-Su,Seo, Jae-Gu,Chung, Myung Jun,Shin, Hea Soon,Ha, Nam Joo The Microbiological Society of Korea 2013 미생물학회지 Vol.49 No.3

프로바이오틱 박테리아는 면역력 활성 조절, 콜레스테롤 수치 억제, 유당내성 강화, 항종양 활성 등의 다양한 생리활성 기능으로 건강 증진 효과가 있는 것으로 보고되고 있다. 프로바이오틱 박테리아는 일단 섭취하게 되면 위장관을 통과하는 동안 산도가 낮거나 단백질분해 효소가 많은 열악한 환경에서 생존해야 하며 프로바이오틱 효과를 발휘하기 위해 증식해야 한다. 이중 코팅 기술은 펩타이드와 다당류의 이중코팅으로 섭취된 프로바이오틱 박테리아를 열악한 조건으로부터 보호하기 위해 개발되었다. 본 연구에서는 이중코팅 된 4종의 비피도박테리움 혼합물의 생존 안정성을 평가하기 위해 코팅이 되지 않은 비피도박테리움 혼합물과 담즙, 산 저항성 및 열 안정성을 비교 평가하였다. 이중 코팅 된 균주와 코팅이 되지 않은 균주를 산과 담즙 조건 및 $40^{\circ}C$에 노출 시킨 후 한천배지에 배양하여 생존생육 세포수를 측정하였으며, BacLigtht 키트를 이용하여 염색 한 후 유세포 분석기를 이용하여 생균과 사균의 세포수를 평가하였다. 이중코팅 된 균주 혼합물의 경우 코팅이 되지 않은 균주 혼합물 보다 산, 담즙 내성이 더 높았으며, 열 안전성 또한 코팅 되지 않은 균주 혼합물보다 높은 것으로 나타났다. 이 같은 결과들로 이중코팅 기술은 프로바이오틱 박테리아의 안정성 및 섭취 후 위장관 트랙을 통과하는 동안 균주의 생존률을 향상시킬 수 있음을 확인하였다. Probiotics have been reported to benefit human health by modulating immunity, lowering cholesterol, improving lactose tolerance, and preventing some cancer. Once ingested, probiotic microorganisms have to survive harsh conditions such as low pH, protease-rich condition, and bile salts during their passage through the gastro-intestinal (GI) tract colonize and proliferate to exert their probiotic effects. The dual coating technology, by which the bacteria are doubly coated with peptides and polysaccharides in consecutive order, was developed to protect the ingested bacteria from the harsh conditions. The aim of the study was to evaluate the viable stability of a doubly coated blend of four species of Bifidobacterium by comparing its bile/acid resistance and heat viability in vitro with that of the non-coated blend. After challenges with acid, bile salts, heat, and viable cell counts (VVCs) of the dual coated and non-coated blend were determined by cultivation on agar plates or flow cytometric measurement after being stain with the BacLigtht kit$^{TM}$. The results showed that the dual coated blend was much higher resistant to the acidic or bile salt condition than the non-coated blend and heat viability was also higher, indicating that the dual coating can improve the survival of probiotic bacteria during their transit through the GI tract after consumption.

Anti-inflammatory and Anti-bacterial Effects of Hyrtios sp. and Haliclona sp.

Sang Hwo Choi(최상화),Su Im Koh(고수임),Seung A Choi(최승아),Hea Soon Shin(신혜순) 환경독성보건학회 2016 한국독성학회 심포지움 및 학술발표회 Vol.2016 No.6