4,7-Dioxobenzo[b]thiophene과 Phthalazine-5,8-dione 유도체 합성 및 항진균 작용 연구

Abstract

Quinones have been reported with various pharmacological activities such as antifungal, anticancer and antimalarial activities. Especially, compounds containing the heterocyclic quinone group represent an important class of biological active compounds. Based on this consideration, methyl 5-((ethoxycarbonyl)(cyano)methyl)-6-chloro-4,7-dihydro-4,7-dioxobenzo[b]thiophene-2-carboxylate derivatives and 5,6-dichloro-2-methyl-1H-benzo[d]imidazole-4,7-dione derivatives were synthesized and evaluated for their antifungal activities. And cinnoline derivatives are widely used in pharmaceuticals and mainly patented as bactericides and fungicides. The inhibitory activity of 6-hydroxycinnolines on the antifungal properties has not been reported to the best of our knowledge. Various 6-hydroxycinnolines with different action and may improve the activities. Heterocyclic quinone compounds represent an important class of biologically active molecules. 5,8-Quinolinedione derivative, a heterocyclic quinone, inhibit cytochrome bc complex by the blockade of mitochondrial electron transport in Saccaromyces cerevisiae, which is different from commonly used antifungal drugs. There have been a few reports on phthalazine-5,8-diones exhibiting cytotoxic activity against cancer cell lines. However, the inhibitory activity of phthalazine-5,8-diones derivatives on the antifungal properties has not been reported to the best of our knowledge. Therefore, phthalazine-5,8-diones with various substituents were designed and synthesized to elucidate their contribution to the antifungal activity. Methyl 5-((ethoxycarbonyl)(cyano)methyl)-6-chloro-4,7-dihydro-4,7-dioxobenzo[b]thiophene-2-carboxylate (PRE-4) was obtained by substitution of methyl 5,6-dichloro-4,7-dihydro-4,7-dioxobenzo[b]thiophene-2-carboxylate with ethylcyanoacetate. Methyl 6-((ethoxycarbonyl)(cyano)methyl)-4,7-dihydro-4,7-dioxo-5-(arylthio)benzo[b]thiophene-2-carboxylate (PRE4-Ss) and 7-Ethyl 2-methyl 6-amino-5,8-dihydro-5-methyl-4,8-dioxo-4H-thieno[2,3-f]indole-2,7-dicarboxylate (PRE-5) were obtained by substitution of PRE-4 with the appropriate arylamines and arylthiols. 3-Ethyl 8-methyl 4-amino-6-hydroxy-5-(arylthio)thieno[3,2-h]cinnoline-3,8-dicarboxylates (PRE7-Ss) were synthesized by treating equivalent amount of hydrazine hydrate with appropriate PRE4-Ss. PYM and PYM-Ss were synthesized via the same method. 6,7-Dichlorophthalazine-5,8-dione (PP-4) and phthalazine-5,8-dione(PP-6) were synthesized by reported methods. 6-Arylaminophthalazine-5,8-diones (PP4-Ns) were synthesized by nucleophilic substitution of PP-4 with appropriate arylamines. 6,7-Bis(arylthio)phthalazine-5,8-diones (PP4-Ss) were synthesized by nucleophilic substitution on PP-4 with two equivalents of appropriate arylthiols. 6-(Phenylamino)phthalazine-5,8-diones (PP6-Ns) were synthesized by nucleophilic substitution of PP-6 with appropriate arylamines. The antifungal activities of all the compounds I have synthesized were evaluated using the two fold broth dilution method against C. albicans, C. tropicalis, C. neoformans, C. krusei, A. flavus and A. niger. Their minimum inhibitory concentration (MIC) values were determined and compared with positive controls, fluconazole and fluorocytosine. Among newly synthesized compounds, methyl 6-((ethoxycarbonyl)(cyano)methyl)-4,7-dihydro-4,7-dioxo-5-(arylthio)benzo[b]thiophene-2-carboxylate (PRE4-Ss) and 7-ethyl 2-methyl 6-amino-5,8-dihydro-5-methyl-4,8-dioxo-4H-thieno[2,3-f]indole-2,7-dicarboxylate (PRE-5) and 3-ethyl 8-methyl 4-amino-6-hydroxy-5-(arylthio)thieno [3,2-h]cinnoline-3,8-dicarboxylates (PRE7-Ss) did not show antifungal activity against all tested pathogenic fungi. Among the other compounds, 6-arylaminophthalazine-5,8-diones (PP4-Ns), 6-(phenylamino)phthalazine-5,8-diones (PP6-Ns) generally showed relatively potent antifungal activities against C. albicans, C. tropicalis, C. neoformans, C. krusei, A. flavus and A. niger compared to fluconazole and fluorocytosine.;Quinonoid 유도체들은 항진균, 항암, 항균, 항말라리아, nitric oxide synthase 저해작용 등 다양한 생리활성을 나타낸다. 본 연구에서는 quinone 유도체 중 우수한 생리활성이 예상되는 methyl 5-((ethoxycarbonyl)(cyano)methyl)-6-chloro-4,7-dihydro-4,7-dioxobenzo[b]thiophene-2-carboxylate, 3-ethyl 8-methyl 4-amino-6-hydroxy-5-(arylthio)thieno[3,2-h]cinnoline-3,8-dicarboxylate, 6,7-dichlorophthalazine-5,8-dione, phthalazine-5,8-dione 유도체를 새로이 합성하여 항진균 작용 등 그 생리활성을 검색하였다. Methyl 5-((ethoxycarbonyl)(cyano)methyl)-6-chloro-4,7-dihydro-4,7-dioxobenzo[b]thiophene-2-carboxylate 유도체들은 2,5-dimethoxybenzaldehyde을 nitration 시킨 후 methylthioglycolate와 반응시킨 후 HCl과 HNO3를 이용해서 산화 및 염소화시켜 Methyl 5,6-dichloro-4,7-dihydro-4,7-dioxobenzo[b]thio phene-2-carboxylate 를 합성하였고, 그 후 ethylcyanoacetate와 반응시켜 Methyl 5-((ethoxycarbonyl)(cyano)methyl)-6-chloro-4,7-dihydro-4,7-dioxobenzo[b]thiophene-2-carboxylate를 합성하였고, 다양한 amine과 arylthiol을 반응시켜, 7-ethyl 2-methyl 6-amino-5,8-dihydro-4,8-dioxo-5-phenyl-4H-thieno[2,3-f]indole-2,7-dicarboxylate (PRE-5,6) 유도체 2개와 methyl 6-((ethoxycarbonyl)(cyano)methyl)-4,7-dihydro-4,7-dioxo-5-(arylthio)benzo[b]thiophene-2-carboxylate (PRE4-Ss) 유도체 4개를 얻었다. 3-Ethyl 8-methyl 4-amino-6-hydroxy-5-(arylthio)thieno[3,2-h]cinnoline-3,8-dicarboxylate 유도체들은 methyl 6-((ethoxycarbonyl)(cyano)methyl)-4,7-dihydro-4,7-dioxo-5-(arylthio)benzo[b]thiophene-2-carboxylate (PRE4-Ss) 유도체에 hydrazine hydrate를 반응시켜 3-ethyl 8-methyl 4-amino-6-hydroxy-5-(arylthio)thieno[3,2-h]cinnoline-3,8-dicarboxylate (PRE7-Ss)유도체 4개를 합성하였다. 이렇게 얻은 cinnoline 유도체들은 tautomerization에 의해 유사 quinone구조를 갖으며 따라서 quinone 계열의 물질들과 마찬가지로 항진균, 항균 작용 때문에 주목 받고 있다. PRE4-Ss 유도체에 hydrazine hydrate를 가하여 구조 내에 질소(N)를 포함하는 유도체인 PRE7-Ss 합성하여 그 항진균 작용을 검색하였다. Phthalazine-5,8-dione 유도체들은 phthalazine을 nitration시킨 후 amination, diamination을 거쳐, 산화와 염소화를 하여 6,7-dichlorophthalazine-5,8-dione(PP-4)를 합성하였다. 얻어진 6,7-dichlorophthalazine-5,8-dione에 3당량의 arylamine 또는 2당량의 arylthiol을 가하여 6-arylamino-7-chlorophthalazine-5,8-diones (PP4-Ns) 유도체 12개와 6,7-bis(arylthio)phthalazine-5,8-dione (PP4-Ss) 유도체 10개를 합성하여 항진균 작용을 검색하였다. 합성한 유도체 (PRE5, PRE6, PRE7, PRE4-Ss, PRE7-Ss, PP4-Ns, PP4-Ss, PP6-Ns)들에 대해서 6가지 병원균주인 C. albicans, C. tropicalis, C. krusei, C. neoformans, A. niger, A. flavus 에 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, fluorocytosine을 사용하였다.