1H-Pyrrolo[3,2-g]isoquinoline-4,9-dione과 pyrrolo[3,2-f]indazole-4,8(1H,7H)-dione 유도체의 합성 및 항진균 작용 연구

Abstract

Quinone 유도체들은 항진균, 항암, 항균, 항말라리아 등 다양한 생리활성을 나타낸다. 본 연구에서는 quinone 유도체 중 우수한 생리활성이 예상되는 다양한 5,8-isoquinolinedione, 4,7-dioxoindazole 유도체를 합성하여 항진균 작용 및 그 생리활성을 검색하였다. 5,6-Dichloro-4,7-dioxoindazole (INDC)을 합성하여 다양한 cyanoacetate 유도체를 합성하여 alkyl 2-(6-chloro-4,7-dihydro-4,7-dioxo-1H-indazol-5-yl) -2-cyanoacetate (INCs) 유도체 4개를 합성하였다. 여기에 aryl amine을 첨가하여 alkyl 6-amino-1,4,7,8-tetrahydro-4,8-dioxopyrrolo[3,2-f]indazole-5-carboxylate (INCAs) 유도체 15개를 합성하였다. 또한 5,6-dichloro-4,7-dioxoindazole (INDC)에 다양한 pyridine 유도체와 반응시켜 중간 유도체인 1-(6-chloro-4,7-dioxo-4,7-dihydro-1H-indazol-5-yl)pyridin-1-ium chloride (INPs) 유도체 2개를 합성하였다. 5,6-Dichloro-4,7-dioxoindazole (INDC)에 다양한 pyridine 유도체를 반응 시킨 후, active methylene compounds를 반응시켜 1H-indolizino[3,2-f]indazole-4,11-dione (INPAs) 유도체 8개를 합성하였다. Isoquinolinedione의 moiety를 포함하는 methyl 5,8-dihydro-1,3-dimethyl-5,8-dioxoisoquinoline-4-carboxylate (KR)을 2',5'-dihydroxyacetophenone과 silver(Ⅱ) oxide, methyl 3-aminocrotonate를 이용하여 합성하였고, 여기에 다양한 pyridine 유도체를 합성하여 1-(4-(methoxycarbonyl)-1,3-dimethyl-5,8-dioxo-5,8-dihydroisoquinolin-7-yl)pyridin-1-ium chloride (KRPs) 유도체 2개를 합성하였다. 5,8-Dihydro-1,3-dimethyl-5,8-dioxoisoquinoline-4-carboxylate (KR)에 다양한 pyridine 유도체를 반응 시킨 후 active methylene compounds를 반응시켜서 methyl 5,12-dihydro-1,3-dimethyl-5,12-dioxoindolizino[2,3-g]isoquinoline-4-carboxylate (KRPAs) 유도체 13개를 얻었다. 신약 개발의 중요한 lead compound인 isoquinolinedione을 다양한 방법으로 oxychlorination시켜 얻은 6,7-dichloroisoquinoline-5,8-dione (IQDC)에 pyridine 유도체와 반응시켜 중간 유도체인 1-(6-chloro-5,8-dioxo-5,8-dihydroisoquinolin-7-yl)pyridin-1-ium chloride (IQPs) 유도체 2개를 합성하였다. 6,7-Dichloroisoquinoline-5,8-dione에 다양한 pyridine 유도체를 반응 시킨 후 active methylene compounds를 반응시켜, indolizino[2,3-g]isoquinoline-5,12-dione (IQPAs) 유도체 11개를 얻었다. 2-((Triphenylphosphin)-ylideneamino)-2H-[1,2,3]triazolo[4,5-g]isoquinoline-4,9-dione (IQPPh)은 6,7-dichloroisoquinoline-5,8-dione (IQDC)에 sodium azide와 triphenylphosphine을 반응시켜 합성하였고, 여기에 benzaldehyde, 10% HCl 수용액, acetone을 각각 반응시켜, 2H-[1,2,3]triazolo[4,5-g]isoquinoline-4,9-dione (IQTAs) 유도체 3개를 합성하였다. 6,7-Dichloroisoquinoline-5,8-dione (IQDC)에 morpholine, thiomorpholine, piperidine을 각각 반응시켜 중간 유도체인 6-chloro-7-(n-phenyl)-isoquinoline-5,8-dione (IQMs) 유도체 3개를 합성하였고, 여기에 sodium azide를 추가로 반응시켜서 3,4-dihydro-1H-[1,4]oxazino[4',3':1,2]imidazo[4,5-g]isoquinoline-6,11-dione derivatives (IQMAs) 유도체를 3개 합성하였다. 6,7-Dichloroisoquinoline-5,8-dione (IQDC)에 다양한 cyanoacetate 유도체를 합성하여 alkyl 2-(7-chloro-5,8-dihydro-5,8-dioxoisoquinolin-6-yl)-2-cyano- acetate (IQCs) 유도체 6개를 합성하였고, 여기에 aryl amine을 반응시켜 alkyl 2-amino-4,9-dihydro-4,9-dioxo-1H-pyrrolo[3,2-g]isoquinoline-3-carboxylate (IQCAs) 유도체 15개를 합성하였다. 합성한 물질 6a-o (INCAs), 7a-b (INPs), 8a-h (INPAs), 11a-b (KRPs), 12a-m (KRPAs), 19a-b (IQPs), 20a-k (IQPAs), 21 (IQPPh), 22-24 (IQTAs), 25a-c (IQMs), 26a-c (IQMAs), 28a-o (IQCAs)의 항진균 작용을 알아보기 위하여 병원성 진균의 대표적인 지표가 되는 6가지 병원균주인 C. albicans, C. tropicalis, C. krusei, C. neoformans, A. niger 및 A. flavus 에 대한 항진균 작용을 검색하였다. 각 화합물에 대한 minimum inhibitory concentration (MIC)는 액체 배지 희석법 (two fold broth dilution method)으로 하였고 대조 약물로는 fluconazole, flucytosine을 사용하였다. 물질 6f (5-C2H5, 4-CF3), 8a (10-H), 12c (6-CH3, 8-H), 20c (6-COCH3, 8-H), 24 (IQTA3), 26c (IQMA3), 28m ((CH2)3CH3, 4-F)이 대부분의 균주에 대해 fluconazole과 flucytosine보다 높거나 비슷한 효과를 나타내어 합성된 유도체중 가장 좋은 항진균 작용을 나타내었다.;Quinones have various pharmacological activities such as antifungal, anticancer and antimalarial activities. Especially, compounds containing the heterocyclic quinone group represent an important class of biologically active compounds. Based on this consideration, 5,8-isoquinolinedione, 4,7-dioxoindazole derivatives were synthesized and evaluated for their antifungal activities. Alkyl 6-amino-1,4,7,8-tetrahydro-4,8-dioxopyrrolo[3,2-f]indazole-5-carbox-ylate (INCs) were synthesised by substitution of 5,6-dichloro-4,7-dioxoindazole (INDC) with cyanoacetate derivatives. And 6-amino-1,4,7,8-tetrahydro-4,8-dioxopyrrolo[3,2-f]indazole-5-carboxylate (INCAs) were synthesised by substitution of alkyl 6-amino-1,4,7,8-tetrahydro-4,8-dioxopyrrolo[3,2-f]indazole-5-carboxylate (INCs) with aryl amines. And intermediate compounds, 1-(6-chloro-4,7-dioxo-4,7-dihydro-1H-indazol-5-yl)pyridin-1-ium chloride (INPs) were synthesised by substitution of 5,6-dichloro-4,7-dioxoindazole (INDC) with pyridine derivatives. And 1H-indolizino[3,2-f]indazole-4,11-dione (INPAs) were synthesised by substitution of 1-(6-chloro-4,7-dioxo-4,7-dihydro-1H-indazol-5-yl)pyridin-1-ium chloride (INPs) with active methylene compounds. Methyl 5,8-dihydro-1,3-dimethyl-5,8-dioxoisoquinoline-4-carboxylate(KR), which contains isoquinoline moiety was obtained by substitution of 2',5'-dihydroxyacetophenone, silver(Ⅱ) oxide and methyl 3-aminocrotonate. And 1-(4-(methoxycarbonyl)-1,3-dimethyl-5,8-dioxo-5,8-dihydroisoquinolin-7-yl)pyridin-1-ium chloride (KRPs) were synthesised by substitution of methyl 5,8-dihydro-1,3-dimethyl-5,8-dioxoisoquinoline-4-carboxylate (KR) with pyridine derivatives. Methyl 5,12-dihydro-1,3-dimethyl-5,12-dioxoindolizino[2,3-g]isoquinoline-4-carboxylate (KRPAs) were synthesised by substitution of 1-(4-(methoxycarbonyl)-1,3-dimethyl-5,8-dioxo-5,8-dihydroisoquinolin-7-yl)pyridin-1-ium chloride (KRPs) with active methylene compounds. 6,7-Dichloroisoquinoline-5,8-dione (IQDC), which is important lead compound of for optimization and drug development was synthesised by multi-ways. And intermediate compounds, 1-(6-chloro-5,8-dioxo-5,8-dihydroisoquinolin-7-yl)pyridin-1-ium chloride (IQPs) were synthesised by substitution of 6,7-dichloroisoquinoline-5,8-dione (IQDC) with pyridine derivatives. And indolizino[2,3-g]isoquinoline-5,12-dione (IQPAs) were synthesised by substitution 1-(6-chloro-5,8-dioxo-5,8-dihydroisoquinolin-7-yl)pyridin-1-ium chloride (IQPs) of with active methylene compounds. 2-((Triphenylphosphin)-ylideneamino)-2H-[1,2,3]triazolo[4,5-g]isoquinoline-4,9-dione (IQPPh) was synthesised by substitution of 6,7-dichloroisoquinoline-5,8-dione with sodium azide and triphenylphosphine. And 2H-[1,2,3]triazolo[4,5-g]isoquinoline-4,9-dione (IQTAs) were synthesised by substitution of benzaldehyde, 10% HCl aqueous solution and acetone. Intermediate compounds, 6-chloro-7-(n-phenyl)-isoquinoline-5,8-dione (IQMs) were synthesised by substitution of 6,7-dichloroisoquinoline-5,8-dione (IQDC) with morpholine derivatives. And 3,4-dihydro-1H-[1,4]oxazino[4',3':1,2]imidazo[4,5-g]isoquinoline-6,11-dione derivatives (IQMAs) were synthesised by substitution 6-chloro-7-(n-phenyl)-isoquinoline-5,8-dione (IQMs) of with sodium azide. Alkyl 2-(7-chloro-5,8-dihydro-5,8-dioxoisoquinolin-6-yl)-2-cyanoacetate (IQCs) were synthesised by substitution of 5,6-dichloro-4,7-dioxoindazole (INDC) with cyanoacetate derivatives. And alkyl 2-amino-4,9-dihydro-4,9-dioxo-1H-pyrrolo[3,2-g]isoquinoline-3-carboxylate (IQCAs) were synthesised by substitution of alkyl 2-(7-chloro-5,8-dihydro-5,8-dioxoisoquinolin-6-yl)-2-cyanoacetate (IQCs) with aryl amines. The antifungal activities of all the synthesized compounds were evaluated using the two fold broth dilution method against C. albicans, C. tropicalis, C. neoformans, C. krusei, A. flavus and A. niger. Their MIC (minimum inhibitory concentration) values were determined and compared with positive controls, fluconazole and flucytosine. Among newly synthesized compounds, 6f (5-C2H5, 4-CF3), 8a (10-H), 12c (6-CH3, 8-H), 20c (6-COCH3, 8-H), 24 (IQTA3), 26c (IQMA3), 28m ((CH2)3CH3, 4-F) generally showed relatively potent antifungal activities compared to fluconazole and flucytosine.