• Archives of Pharmacal Research
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• 제17권6호
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• pp.420-423
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• 1994

Various N'-substituted anilino-N-methyl-N'-nitrosoureas(2a-n) were easily prepared from the reaction of substituted phenylhydraines $(3, 4-CH_3, {\;} 3-, {\;} 4-OCH_3, {\;} 3-, {\;} 4-F, {\;} 3, {\;} 4-Cl, {\;} 4-Br, {\;} 2-, {\;} 3-, {\;} 4-NO_2, 4-(NO_2)_2)$ with methyl isocyanate, followed by the nitrosation with 99% HCOOH and dry sodium lnitrite powder. Surprisingly, of these series of analogus, the anilino-nitrocosureas substituted with eletron-withdrawing nitro groups (2k-a) showed significantly low $ED_{30}$ values of $1.4-3.4 {\mu}g/ml.$ In addition, none of these copounds subtituted with electron-donating groups exhibited cytotoxicities.

• 한국융합학회논문지
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• 제13권1호
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• pp.161-166
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• 2022

HIF-2α는 저산소 조건에서 활성화되는 전사인자로 암, 대사증후군, 관절염, 간염 등의 발병 기전에서 핵심 역할을 한다고 보고된 바 있다. 이에 HIF-2α 저해제를 도출하고자 기존 약리활성 구조를 도입한 N'-arylisonicotinolyhydrazide를 골격으로 설정하고 화합물 라이브러리로부터 해당 화합물들을 선택하여 HIF-2α 저해활성을 측정하였다. 이를 위해 HRE-luciferase를 HTB-94세포에 transfection하고 아데노바이러스를 이용하여 HIF-2α를 세포 내로 도입하여 luciferase reporter gene assay를 수행하였다. 2-hydroxy-1-naphthyl 기를 포함한 화합물에서 저해활성이 발견됨에 따라 이 구조를 포함하는 골격을 다시 설정하고 해당 화합물들을 선정하여 활성을 측정하였다. 그 결과 HIF-2α 저해활성과 위양성 시험을 통하여 2 종의 HIF-2α 저해제를 도출하였다. 본 연구는 생물학과 화학의 융합연구로 수행되었으며 도출된 저해제는 후속 저해제 탐색 연구와 HIF-2α의 기능 연구에 활용될 수 있고 관련 질환의 치료제 개발에도 기여 할 것으로 사료된다.

• Tuberculosis and Respiratory Diseases
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• 제43권1호
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• pp.46-53
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• 1996

연구배경: 폐포대식세포가 내독소에 노출되면 이 후 자극에 의해 종양괴사인자, 활성산소 등 독성이 강한 분비물 방출이 더욱 촉진되어 성인성호흡곤란증후군과 같은 각종 폐질환이 초래되는 중요한 기전으로 이해되고 있다. 그러나 문헌에 보고된 내독소에 의한 이런 priming효과는 다형핵백혈구, 단핵세포나 동물의 폐포대식세포를 대상으로 한 결과로서, 인체 폐포내에서의 폐포대식세포, 폐포상피세포 및 내독소의 상호작용을 밝히는 면에서는 한계가 있었다. 방법: 폐포상피세포에서 기원한 폐암세포주인 A549 24-well Linbro plate에 배양하여 세포단층을 형성한 후 기관지폐포세척술로 얻은 사람의 폐포대식세포($10^6/ml$)를 A549세포에 부착시켜 생체내와 유사한 환경을 만들어 실험을 시행하였다. 방법은 A549세포 부착 전후에 내독소(500 ng/ml)로 전처치한 다음 PMA, fMLP로 자극하여 방출된 과산화수소를 nM/well/2hrs 단위로 측정하여 대조군과 비교하였고 아울러 A549세포만종 없이 각 well에 직접 폐포대식세포단층을 형성한 후 동일하게 처치하였으며 결과는 다음과 같았다. 결과: 1) 24-well의 표면에 폐포대식세포단층을 형성한 후 내독소로 처치(n=7)한 경우, 대조군은 무자극군, PMA 자극군, fMLP자극군에서 각각 $9.48{\pm}2.44$, $10.38{\pm}2.34$, $10.28{\pm}2.33$ nM/well/2hrs의 과산화수소 분비능을 보였고 내독소 전처치군은 각각 $9.56{\pm}2.15$, $9.82{\pm}1.80$, $11.31{\pm}2.48$ nM/well/2hrs을 보여 내독소에 의한 priming효과는 없었다. 2) 폐포대식세포를 미리 내독소로 처치한 후 A549세포단층에 부착(n=7)시킨 경우에는 대조군의 과산화수소 분비능은 무자극군 $4.82{\pm}1.59$, PMA자극군 $8.31{\pm}1.67$, fMLP자극군 $7.06{\pm}1.82$ nM/well/2hrs이고 내독소 전처치군은 각각 $4.44{\pm}1.41$, $7.05{\pm}1.64$, $6.32{\pm}1.69$ nM/well/2hrs로서 내독소에 의한 priming 효과는 없었다. 3) 폐포대식세포를 A549세포단층에 부착시킨 후 내 독소로 처치(n=11)하면 대조군은 무자극군, PMA자극군, fMLP자극군에서 각각 $4.33{\pm}1.04$, $7.94{\pm}1.42$, $6.00{\pm}1.22$ nM/well/2hrs의 과산화수소를 분비하였고 내독소 전처치군은 각각 $5.43{\pm}1.19$, $7.56{\pm}1.31$, $6.38{\pm}1.19$ nM/well/2hrs를 보여 A549세포 부착후에도 내독소의 priming 효과는 관찰되지 않았으나 PMA나 fMLP로 자극하지 않은 무자극군에서 내독소 전처치군이 대조군에 비해 과산화수소 분비능의 유의한 증가를 보였다(p<0.05). 결론: 이상의 결과는 사람 폐포대식세포에서 내독소에 의한 priming효과는 없으나 폐포상피세포와의 상호작용에 의해 내독소가 폐포대식세포를 직접 자극함을 시사하는 소견이며 향후 추시가 필요할 것으로 사료된다.

• Archives of Pharmacal Research
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• 제17권3호
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• pp.135-138
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• 1994

Acyclonucleoside homologues of 1-$(\omega$-cyabnoalkyl)-and 1, 3-bis $(\omega$-cyanoalkyl) uracils were synthesized by the series of alkylation reactions of uracil with the $\omega$-chloroalkyl nitrile ${(Cl-(CH}_2)_n$-CN;n=1, 2, 3, 4) in DMSO under $50-70^circ{C}$ temperature. The 1-$(\omega$-cyanoalkyl)-and 1, 3-bis$(\omega$-cyanoalkyl) uracils were separated either by the fractional crystallization or column chromatography. The antitumor activities for these synthesized compounds were determined against four cell lines (J-82 cell, P388 cell, FM-3A cell and U-937 cell lines). These compounds failed to exhibit any significant antitumor activity.

• Biomolecules & Therapeutics
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• 제14권3호
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• pp.143-147
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• 2006

In the mouse leukemic monocyte cell line RAW 264.7, the vacuolar-type $(H^+)$-ATPase (V-ATPase) inhibitor bafilomycin $A_1$ at 10 and 100 nM decreased cell growth and survival as determined by 3-(4,5-dimethyl(thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in a concentration-dependent manner. At such concentrations, bafilomycin $A_1$ induced nitric oxide (NO) production through the expression of inducible nitric oxide synthase (iNOS). The bafilomycin $A_1$-induced NO production was inhibited by the NOS inhibitor $N^G$-monomethyl-L-arginine acetate (L-NMMA). Our findings suggest that the V-ATPase inhibitor bafilomycin $A_1$ induces NO production through the expression of iNOS protein.

• Asian-Australasian Journal of Animal Sciences
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• 제22권4호
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• pp.492-499
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• 2009

A Silkie Bantam embryo fibroblast line (named SBF59 line) was successfully established by using direct explant culture and cryopreservation techniques. Cell morphology, viability, dynamic growth and contamination were tested and the karyotype and levels of isoenzymes of lactic dehydrogenase and malic dehydrogenase were analyzed. Four kinds of fluorescent protein extrogenes, including $pEGFP-N_3$, $pECFP-N_1$, $pEYFP-N_1$ and $pDsRed1-N_1$ were transfected into the cells. The results showed that the cells were healthy and possessed a fibrous structure without a change in morphology. The average viability of the cells was 96% before freezing and 90.5% after thawing. The growth curve appeared as typical "S" shape and the cell growth passed through a detention phase, a logarithmic phase and a platform phase; the estimated population doubling time (PDT) was 38.5 h; assays for the presence of bacteria, fungi, viruses and mycoplasmas were negative; the cell line showed no cross contamination when assessed by isoenzyme analysis; the chromosome number was 2n = 78 on more than 88% of occasions; four kinds of fluorescent protein extro-genes appeared to be expressed effectively with a high transfection efficiency between 18.3% and 42.3%. The cell line met the required quality control standard. It not only preserves the genetic resources of the important Silkie Bantam at the cellular level but also provides valuable materials for genomic, post-genomic, somatic cell cloning research and other applications.

• Preventive Nutrition and Food Science
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• 제19권4호
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• pp.268-273
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• 2014

The induction of detoxification enzymes by benzyl isothiocyanate (BITC) and its synthetic N-acetyl-L-cysteine (NAC) conjugate (NAC-BITC) was examined in Hepa1c1c7 murine hepatoma cells. BITC and NAC-BITC inhibited Hepa1c1c7 cell growth in a dose-dependent manner. Cell growth was 4.5~57.2% lower in Hepa1c1c7 cells treated with $0.1{\sim}1.0{\mu}M$ BITC than in control-treated Hepa1c1c7 cells. The NAC-BITC treatment had a similar inhibitory pattern on Hepa1c1c7 cell growth; $0.5{\mu}M$ and $10{\mu}M$ NAC-BITC decreased cell growth by 13.6% and 47.4%, respectively. Treatment of Hepa1c1c7 cells with $0.1{\sim}2.0{\mu}M$ BITC also elicited a dose-response effect on the induction of quinone reductase quinone reductase (QR) activity and QR mRNA expression. Treatment with $1{\mu}M$ and $2{\mu}M$ BITC caused 1.8- and 2.8-fold inductions of QR mRNA, respectively. By comparison, treatment with $1{\mu}M$ and $2{\mu}M$ NAC-BITC caused 1.6-and 1.9-fold inductions of QR mRNA, respectively. Cytochrome P450 (CYP) 1A1 and CYP2E1 induction were lower in $0.1{\sim}2{\mu}M$ BITC-treated cells than in control-treated cells. CYP2E1 activity was 1.2-fold greater in $0.1{\mu}M$ NAC-BITC-treated cells than in control-treated cells. However, the CYP2E1 activity of cells treated with higher concentrations (i.e., $1{\sim}2{\mu}M$) of NAC-BITC was similar to the activity of control-treated cells. Considering the potential of isothiocyanatesto prevent cancer, these results provide support for the use of BITC and NAC-BITC conjugates as chemopreventive agents.

• 대한수의학회지
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• 제63권1호
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• pp.3.1-3.9
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• 2023

The efficiency of somatic cell nuclear transfer (NT) in pigs is low and requires enhancement. We identified the most efficient method for zona pellucida (ZP) removal and blastomere aggregation in pigs and investigated whether the aggregation of NT and parthenogenetic activation (PA) of blastomeres could reduce embryonic apoptosis and improve the quality of NT-derived embryos by investigating. Embryonic developmental competence after ZP removal using acid Tyrode's solution or protease (pronase E). The embryonic developmental potential of NT-derived blastomeres was also investigated using well-of-the-well or phytohemagglutinin-L. We analyzed apoptosis in aggregate-derived blastocysts. The aggregation rate of protease-treated embryos was lower than that of Tyrode's solution-treated embryos (69.2% vs. 88.3%). No significant difference was observed between phytohemagglutinin-L and well-of-the-well (35.7%-38.5%). However, 2P1N showed a higher number of blastocysts compared to 3N (73.8% vs. 24.3%) and an increased blastocyst diameter compared to the control and 1P2N (274 ㎛ vs. 230-234 ㎛). In blastomeres aggregated using phytohemagglutinin-L, the apoptotic cell ratio was significantly higher in 1P2N and 3N than in 3P (5.91%-6.46% vs. 2.94%, respectively). Our results indicate that aggregation of one NT embryo with two PA embryos improved the rate of blastocysts with increased blastocyst diameter.

• Journal of Microbiology and Biotechnology
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• 제7권5호
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• pp.329-332
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• 1997

Cell-free extracts of Streptomyces neyagawaensis SL-387 grown on a chemically defined medium supplemented with DL-3-amino-3-phenylpropionic acid (APP) produced N-acetyl-APP (Ac-APP) in the presence of APP and acetyl coenzyme A. The APP obtained by acid hydrolysis of the Ac-APP was D-configuration: $[\alpha]_D+6.5^{\circ}(H_2O)\;at\;20^{\circ}C$, optical purity 92% enantiomeric excesses (ee). These results suggest that an N-acetyltransferase exists in the cell-free extract as a novel enzyme with specificity for D-APP.

• 한국수소및신에너지학회논문집
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• 제35권2호
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• pp.162-167
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• 2024

Fossil fuels have been main energy source to people. However, enormous amount of CO₂ was emitted over the world , resulting in global climate crisis today. Recently, solid oxide electrolyzer cell (SOEC) is getting attention as an effective way for producing H₂, a clean energy resource for the future. Also, SOEC could be applicable to reverse water-gas shift reaction process due to its high-temperature operating condition. Here, SOEC system was utilized for both H₂ production and CO₂ reduction process, allowing product gas composition change by controlling operating conditions.