Quinoxaline-5,8-dione 화합물과 1,3,4-trisubstituted pyrazole 화합물의 합성 및 생리활성 연구

Abstract

Quinones have been reported with various pharmcological activities such as antifungal, anticancer and antimalarial activities. Especially, compounds containing the heterocyclic quinone group represent an important class of biologically active molecules. However, the inhibitory activity of quinone classes on the proliferation of SMCs has not been reported to the best of our knowledge. Therefore, I synthesized and tested various quinone derivatives to elucidate their contribution to the antiproliferative effects on PDGF-stimulated SMC proliferation. Among the quinones tested, 6-arylamino-quinoxaline-5,8-dione derivatives (QNXs, J1s, J3s) showed the potent antiproliferative activity on the SMCs. 6-Arylamino-7-chloro-2,3-di(pyridin-2-yl)quinoxaline-5,8-dione (J1s), 6,7- bis-arylthio-2,3-di(pyridin-2-yl)quinoxaline-5,8-dione (J2s), 6-Arylamino- 2,3-di(thiophen-2-yl)quinoxaline-5,8-dione (J3s) and 6-arylthio-2,3-di(thio- phen-2-yl)quinoxaline-5,8-dione (J4s) were obtained by the substitution of quinoxaline-5,8-dione with the appropriate arylamines and arylthiols. The most active potential among the series (QNXs, J1s, J2s, J3s, J4s) was found for the compounds (QNXs) which showed half maximal inhibition of the PDGF-stimulated proliferation of the RAoSMCs tested at the level of 0.4-0.5 M. The results indicate that the 6-arylamino-2,3- dimethyl-quinoxaline-5,8-dione (QNXs) are a promising lead for the development as inhibitors of the SMC proliferations. Pyazole derivatives have been reported with various pharmcological activities such as antianxiety, anticancer, antifungal and antibacterial activities. Among the pyrazole derivatives, I synthesized and tested 1,3,4-trisubstituted pyazoles derivatives to elucidate their biological effects. (1,3-Disubstituted-1H-pyrazol-4-yl)methanols (OCOHs) were obtained by oxidation of 1,3-Disubstituted-1H-pyrazole-4-carbaldehydes (OCs) with LiAlH4. 2-(1,3-Disubstituted-1H-pyrazol-4-yl)-1H-benzoimidazoles, 2-(1,3- disubstituted-1H-pyrazol-4-yl)-benzothiazoles (OCAs, OCNs) were synthe- sized by reaction OCs with the appropriate benzene-1,2-diamines or 2- aminobenzenethiols. [(1,3-Disubstituted-1H-pyrazol-4-yl)methylene]benzo- hydrazides (OCHYs) were obtained by the substitution of OCs with benzoic hydrazide. 1-[2-(1,3-Disubstituted-1H-pyrazol-4-yl)-5-phenyl- [1,3,4]oxadiazol-3-yl]-ethanones (OCCYs) were synthesized by cyclization OCHYs with Ac2O. The antifungal activities of OCAs and OCNs were evaluated using the two fold broth dilution method against C. albicans, C. tropicalis, C. krusei and A. niger. The minimum inhibitory concentration (MIC) values of test compounds were determined, and compared with positive controls, fluconazole and fluorocytosine. Among newly synthesized compounds, 2-[3-(4-chlorophenyl)-1-phenyl-1H-pyrazol-4-yl]-1H-benzo[d]imidazole (OCA2-3), 2-[3-(4-fluorophenyl)-1-phenyl-1H-pyrazol-4-yl]-1H-benzo[d] imidazole (OCA3-2) generally showed relatively potent antifungal activities compared to fluconazole and flucytosine against C. albicans, C. tropicalis, and C. krusei.;가. Quinoxaline-5,8-dione 화합물 합성 Quinone 유도체들은 항진균, 항암, 항균, 항말라리아, nitric oxide synthase 저해작용 등 다양한 생리활성을 나타낸다. 본 연구에서는 quinone 유도체 중 우수한 생리활성이 예상되는 quinoxaline-5,8-dione 유도체를 새로이 합성하여 그 생리활성을 검색하였다. 동맥경화와 혈관재협착의 원인이 되는 혈관평활근세포 (SMCs)의 증식 억제 작용을 갖는 quinone체들은 아직까지 알려져 있지 않다. 따라서 quinoxaline-5,8-dione 유도체에 대한 SMCs의 증식을 저해하는 작용을 검색하였으며 항진균작용도 연구하였다. 2,3-Di(pyridin-2-yl)quinoxaline-5,8-dione 유도체들은 1,4-dimethoxyben- zene을 니트로화한 후 Pt₂O을 촉매로 수소 환원시키고 2,2'-pyridil과 반응시켜 5,8-dimethoxy-2,3-di(pyridin-2-yl)quinoxaline을 합성하였다. 그 후 HCl과 HNO3을 사용하여 quinone form으로 산화 및 염소화시켜 6,7-dichloro-2,3- di(pyridin-2-yl)quinoxaline-5,8-dione을 합성 후 다양한 arylamine과 arylthiol을 반응시켜 6-arylamino-7-chloro-2,3-di(pyridin-2-yl)quinoxaline-5,8-dione (J1s) 유도체 11개와 6,7-bis-arylthio-2,3-di(pyridin-2-yl)quinoxaline-5,8- dione (J2s)유도체 8개를 얻었다. 2,3-Di(thiophen-2-yl)quinoxaline-5,8-dione 유도체들은 위와 마찬가지로 1,4-dimethoxybenzene을 니트로화한 후 Pt₂O을 촉매로 수소 환원시키고 2,2'-thenil과 반응시켜 5,8-dimethoxy-2,3-di(thiophen-2-yl)-5,8-dihydro- quinoxaline을 합성하였다. 그 후 CAN을 사용하여 quinone form으로 산화하여 2,3-di(thiophen-2-yl)quinoxaline-5,8-dione을 합성 후 다양한 arylamine과 arylthiol을 반응시켜 6-arylamino-2,3-di(thiophen-2-yl)quinoxaline-5,8-dione (J3s) 유도체 4개와 6-arylthio-2,3-di(thiophen-2-yl)quinoxaline-5,8-dione (J4s) 유도체 2개를 얻었다. 나. 1,3,4-Trisubstituted pyrazole 화합물 합성 항불안, 항암, 항진균, 살충작용과 항균작용 등 다양한 생리 활성을 나타내는 pyrazole 유도체 중 우수한 생리활성이 예상되는 1,3,4-trisubstituted pyarzole 유도체를 합성하여 그 생리활성을 검색하였다. 1,3,4-Trisubstituted pyazole 유도체들은 세 종류의 acetophenone (4-H, Cl, F)에 phenylhydrazine 또는 2-hydrazinopyridin을 반응하여 1-phenyl-2-(1- phenylethylidene)hydrazines와 1-(1-phenylethylidene)-2-(pyridin-2-yl)hyd- razines를 합성한 후 Vilsmeier-Haack complex (POCI₃/DMF)를 이용하여 여섯 종류의 1,3-disubstituted-1H-pyrazole-4-carbaldehyde (OCs)를 합성하였다. 각각의 OCs를LiAIH₄로 환원하여 (1,3-disubstituted-1H-pyrazol-4-yl) methanol (OCOHs) 유도체 5개를 얻었다. 각각의 1,3-disubstituted-1H-pyrazole-4-carbaldehyde (OCs)에 당량의 2-aminobenzenethiol, 2-amino-4-chloro-benzenethiol 또는 benzene-1,2- diamine, 4,5-dichloro-benzene-1,2-diamine를 반응하여 2-(1,3-disubstituted- 1H-pyrazol-4-yl)-1H-benzoimidazole, 2-(1,3-disubstituted-1H-pyrazol-4-yl) -benzothiazole (OCAs와 OCNs) 유도체 17개를 얻었다. 마지막으로 각각의 1,3-disubstituted-1H-pyrazole-4-carbaldehydes (OCs)에 당량의 benzoic hydrazide를 넣어 [(1,3-disubstituted-1H-pyrazol-4-yl) methylene]benzohydrazide (OCHYs) 유도체 6개를 합성한 후 acetic anhydride로 cyclization하여 1-[2-(1,3-Disubstituted-1H-pyrazol-4-yl)-5- phenyl-[1,3,4]-oxadiazol-3-yl]-ethanone (OCCYs) 유도체 3개를 얻었다. 다. 생리활성 평가 합성된 유도체 (J1s, J2s, J3s, J4s, QNXs, OCAs, OCNs)에 대해서는 SMCs의 증식 저해 작용을 검색하였다. WST colorimeric assay에 의해 각 화합물의 IC50 값을 기준물질인 mycophenolic acid (MPA)와 비교하여 결정하였다. SMCs의 증식 저해작용을 검색한 결과 대부분의 quinoxaline-5,8-dione 유도체들은 대체로 좋은 효과를 나타냈다. 특히 QNXs와 J1s, J3s의 대부분의 물질들이 우수한 활성을 보여주었으며 이들 물질의 활성은 MPA보다 우수하거나 비슷하였다. 반대로 물질 J2s, J4s와 OCAs, OCNs는 우수한 활성을 나타내지 않았다. 합성한 유도체 (J1s, J2s, OCAs, OCNs)들에 대해서는 4가지 병원균주인 Candida albicans, C. tropicalis, C. krusei, Aspergillus niger에 대해서 항진균 작용을 검색하였다. 각 화합물에 대한 MIC (minimum inhibitory concentration)는 액체 배지 희석법 (two fold broth dilution method)으로 하였고 대조 약물로는 fluconazole, flucytosine을 사용하였다. 대부분의 유도체들이 C. krusei에서는 비교적 효과가 좋았지만, 다른 균주에는 대체로 효과가 좋지 않은 것으로 나타났다. Pyrazole 모핵의 4번 위치에 benzoimidazole이 치환된 물질인 OCA2-3, OCA3-3는 C. krusei 뿐만 아니라 C. albicans과 C. tropicalis 균주에서도 모두 fluconazole과 flucytosine 보다 뛰어난 항진균 효과를 보였다.