Indolequinone과 6-hydroxycinnoline 화합물의 합성 및 항진균 작용 연구

Abstract

Quinonoid 유도체들은 항진균, 항암, 항균, 항말라리아, nitric oxide synthase 저해작용 등 다양한 생리활성을 나타낸다. 본 연구에서는 quinone 유도체 중 우수한 생리활성이 예상되는 1H indole-4,7-dione, (5E,6E)-5,6-bis(benzylideneamino)-3 chlorocyclohexa-2,5-diene-1,4-dione 및 6 hydroxycinnoline, 유도체를 새로이 합성하여 그 생리활성을 검색하였다. 암세포에 대한 세포독성 작용이 우수하다고 알려 indolequinone 모핵에 당량 혹은 과량의 arylthiol을 넣고 치환 반응을 시켜 YOIBs 유도체 12개와 YOIs 유도체 2개를 합성하였다. 또한 3,6-dimethoxy-1,2-dimethylcyclohexa -1,4-diene에 oxychlorination을 하여 2,3-dichloro-5,6-dimethylcyclohexa- 2,5-diene-1,4-dione를 합성한 후 methyl cyanoacetate를 반응시켜 methyl기와 cyano acethyl 기가 치환된 새로운 indolequinone모핵인 methyl-2-(2-chloro-4,5-dimethyl-3,6-dioxocyclohexa-1,4-dienyl)-2-cyano- acetate를 합성하였다. 그 후 당량의 methylamine 또는 arylamine을 첨가하여 YOMNs 유도체 11개와 YOM-CM, YOM-CE를 합성하였다. 신약 개발의 중요한 lead compound인 isoquinolinedione과 quinolinedione을 chlorination하고 methyl cyanoacetate를 반응시킨 후 arylamines을 첨가하여 분자내 고리화 반응에 의해heterocyclic indoledione구조를 갖는 IQNs 와 YQNs를 합성하였다. 5-Methoxy-1H-indole-3-carbaldehyde를 출발 물질로 하여 새로운 모핵인 3-(hydroxymethyl)-5-methoxy-1-methyl-1H indole-4,7-dione(YID)와 4,7-dihydro-5-methoxy-1-methyl 4,7-dioxo-1H-indole-3-carbaldehyde(YIC)를 얻은 후 당량의 arylthiols을 넣고 YIDNs 유도체 12개와 YICNs 유도체 2개를 합성하였다. Cinnoline 유도체들은 tautomerization에 의해 유사 quinone구조를 갖으며 따라서 quinone 계열의 물질들과 마찬가지로 항진균, 항균 작용 때문에 주목 받고 있다. 다양한 모핵에 hydrazine hydrate를 가하여 구조 내에 질소(N)를 포함하는 6 hydroxycinnoline 유도체인 IQH, YQH, YOM-H, YOM-TCs 유도체 5개를 합성하여 그 항진균 작용을 검색하였다. 합성한 유도체 (YOIBs, YOIs, YOM-CE, YOM-CM, YOMNs, IQNs, YQNs, YICNs, YIDNs, JNN, YOM-TCs, YOM-H, IQH, YQH)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. YOIBs 와 YOIs 중 대부분의 유도체에서 C. neoformans 균주에 대해서 거의 모든 유도체가 fluorocytosine 보다 좋거나 비슷한 효과를 나타내었으며, 나머지 5개 균주에 대해서는 일부를 제외한 거의 대부분의 유도체가 fluconazole 보다 효과가 좋지 않았다. IQNs 유도체 중에서는 특히 IQN1(4-CH3), IQN2(4-F)가 C. krusei와 C. neoformans, C. tropicalis 균주에서 비교적 효과가 좋았지만 그 외 IQNs 유도체와 YQNs 유도체는 대체로 효과가 좋지 않은 것으로 나타났다. YOM-CM은 C. krusei와 C. neoformans 균주에 대해서 fluorocytosine 보다 좋은 효과를 보였으나 YOM-CE는 모든 균주에서 fluorocytosine보다는 효과가 약하였다. YOMNs 중에서 일부 유도체가 C. krusei와 C. neoformans, A. niger 균주에서 비교적 효과가 좋았으나 A. flavus에는 전혀 효과가 없었다. C. tropicalis, C. albicans 균주에서는 각각 YOMN8(4-Br), YOMN1(4-H)만이 유일하게 fluorocytosine 보다 좋은 효과를 보였다. 한편 YICNs, YIDNs, JNNs 유도체들은 모든 균주에서 대체로 효과가 좋지 않은 것으로 나타났다. 또한 6-hydroxycinnoline 유도체들에 대해서도 항진균 작용을 검색해 본 결과, YOM-H는 A. niger, YOM-TC1(4-H), YOM-TC2(4-CH3), YOM-TC3(3,4-CH3)는 C. krusei와 C. neoformans, YOM-TC4(3-F)는 C. neoformans와 A. niger, YOM-TC5(3-CH3)는 C. krusei 균주에 대해서 fluorocytosine보다 좋은 효과를 나타내었다. IQH는 C. neoformans, C. krusei와 C. tropicalis에 대해서는 fluorocytosine와 비슷하게 항진균 작용을 나타내었으나 YQH는 C. tropicalis, A. niger를 제외한 4가지 균주에서 fluconazole 보다는 효과가 좋았다.;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 biologically active compounds. Based on this consideration, indolequinone, and (5E,6E)-2-arylamino-5,6-bi (benzylideneamino)-3-chlorocyclohexa-2,5- diene-1,4-diones derivatives were synthesized and evaluated for their antifungal activites. 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 substituents could exhibit the biological activities through different action and may improve the activities. 5,6-Bis(arylthio)-1H-indole-4,7-dione (YOIBs) and 5-(arylthio)-6-chloro-1H-indole-4,7-dione (YOIs) were obtained by substitution of 1H-indole-4,7-dione (YOI-A) with the appropriate arylthiols. Methyl 4,7-dihydro-1,5,6-trimethyl-4,7-dioxo-1H-indole-3-carboxylate (YOM-CM), methyl 4,7-dihydro-5,6-dimethyl-4,7-dioxo-1-arylamino-1H- indole-3-carboxylate (YOMNs) were synthesized by reaction of YOM-BM with appropriate amines, ethyl 4,7-dihydro-1,5,6-trimethyl-4,7-dioxo-1H- indole-3-carboxylate (YOM-CE) were synthesized from YOM-BE. IQ was synthesized by treating HCl and HNO3 with isoquinoline, and IQ-A was obtained via intramolecular cyclization by treating methyl cyanoacetate. YQ and YQ-A were synthesized from quinoline via the same method. A variety of 6-hydroxycinnolines were synthesized by treating equivalent amount of hydrazine hydrate with appropriate starting compounds, including IQ-A, YQ-A, YOM-BM, YOM-Ts. The products of cyclization for each compounds are methyl 3-amino-5-chloro-6-hydroxypyrido[3,4-h]cinnoline-4-carboxylate (IQH), methyl 3-amino-5-chloro-6-hydroxypyrido [2,3-h]cinnoline-4-carboxylate (YQH), methyl 3-amino-5-chloro-6,8a-dihydro-7,8-dimethyl-6-oxocinnoline-4-carboxylate (YOM-H), methyl 5- aryltiol-3-amino-6-hydroxy-7,8-dimethylcinnoline-4-carboxylate(YOM-TCs). 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, 2-arylamino-5,6-bis(benzylidene amino)-3-chlorocyclohexa-2,5-diene-1,4-diones(JNNs), 5-arylamino-3-(hydroxymethyl)-1-methylindole-4,7-dione (YICNs) and 5-arylamino-4,7 -dihydro-1-methyl-4,7-dioxo-1H-indole-3-carbaldehyde (YIDNs) did not show antifungal activity against all tested pathgenic fungi. Among the other compounds, methyl-4,7-dihydro -5,6-dimethyl-4,7-dioxo-1-arylamino- 1H-indole-3-carboxylate (YOMNs), 6-hydroxycinnolines (YOM-H, YOM-TCs, IQH, YQH) generally showed relatively potent antifungal activities against C. tropicalis, C. krusei, C. neoformans and A. niger .compared to fluconazole and fluorocytosine.