6-Hydroxybenzocinnoline 화합물과 1,3,4-trisubstituted pyrazole 화합물의 합성 및 항진균 작용 연구

Abstract

Pyazole derivatives have been reported with various pharmcological activities such as antianxiety, anticancer, antifungal and antibacterial activities. Among the pyrazole derivatives, 1,3,4-trisubstituted pyazole derivatives were synthesized and tested to elucidate their biological effects. (1,3-Disubstituted-1H-pyrazol-4-yl)methanols 4a-d (JERs) were obtained by oxidation of 1,3-disubstituted-1H-pyrazole-4-carbaldehydes 3a-d(JEAs) with LiAlH4. 2-(1,3-Disubstituted-1H-pyrazol-4-yl)-1H-benzoimidazoles, 2-(1-disubstituted-1H-pyrazol-4-yl)-benzothiazoles 5a-1(JEs) derivatives were synthesized by reaction JEAs with the appropriate benzene-1,2-di-amines or 2-aminobenzenethiols. [(1,3-Disubstituted-1H-pyrazol-4-yl)methylene]benzohydrazides 6a-d (JEBHs) were obtained by the substitution of JEAs with benzoic hydrazide. 1-[2-(1,3-Disubstituted-1H-pyrazol-4-yl)-5-phenyl-[1,3,4]-oxadiazol-3-yl]-ethanones 7a-d (JECYs) were synthesized by cyclization of JEBHs with Ac₂O. 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. 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, 2,3-dichloro-naphthalene-1,4-dione (8) derivatives, indolequinones, 6-methyl-2H-chromene-2,5,8-trione (23) (COU) derivatives were synthesized and evaluated for their antifungal activites. Methyl 2-(2-chloro-1,4-dihydro-1,4-dioxonaphthalen-3-yl)-2-cyanoacetate (9) (JQM) was obtained via intramolecular cyclization by treating 2,3-di-chloronaphthalene-1,4-dione (8) with methyl cyanoacetate. Ethyl 2-(2-chloro-1,4-dihydro-1,4-dioxonaphthalen-3-yl)-2-cyanoacetate (14) (JQE) was synthesized from 2,3-dichloronaphthalene-1,4-dione (8) via the same method. 2-[2-Arylthio-1,4-dihydro-1,4-dioxo-naphthalen-3-yl]-2-cyanoacetate 10a-d (JQMSs) and ethyl 2-(2-arylthio-1,4-dihydro-1,4-dioxonaphthalen-3-yl)-2-cyanoacetate 15a-f (JQESs) were obtained by the substitution of JQM and JQE with appropriate arylthiols. A variety of 6-hydroxycinnolines were synthesized by treating equivalent amount of hydrazine hydrate with appropriate starting compounds, including (9) (JQM), 10a-d (JQMSs) and (14) (JQE). The products of cyclization for each compounds are methyl 3-amino-6-hydroxy-5-(phenyl-thio)benzo[h]cinnoline-4-carboxylate (11) (JQMCY1), 5-arylthio-3-amino-6-hydroxy-benzo[h]cinnoline-4-carboxylate 12a-c (JQMCY2-4) and ethyl 3-amino-5-chloro-4a,10b-dihydro-6-hydroxybenzo[h]cinnoline-4-carboxylate (16) (JQECY). 2-(3-Chloro-4-hydroxy-1-oxonaphthalen-2(1H)-ylidene)malononitrile (17) (JQN) was prepared by treating malononitrile with 2,3-dichloronaphthalene-1,4-dione (8). 2-[1,4-Dihydro-1,4-dioxo-2-(phenylthio)naphthalen-3-yl] malononitrile (18) (JQNS) and 2-amino-4,9-dihydro-4,9-dioxo-1-substituted-1H-benzo[f]indole-3-carbonitrile 19a-e (JQNNs) were obtatined by sub-stitution of arylthiols or arylamines in the presence of CeCl₃. A new compound 6-methyl-2H-chromene-2,5,8-trione (23) (COU) was prepared by treating 6-methylcoumarin (20) with nitration, reduction and oxidation by Fremy's salt. 6-Methyl-7-arylthio-2H-chromene-2,5,8-triones 24a-j (COSs) derivatives were obtained by sustitution of (23) (COU) with the appopriate arylthiols. The antifungal activities of all the compounds 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, (15g) (JQES7) showed relatively potent antifungal activites compared to fluorocytosine against C. albicans, C .tropicalis and especially C. krusei. (15d) (JQES4) showed similar antifungal activity with fluorocytosine against A. niger. (12d) (JQMSCY4) showed potent antifungal activities compared to fluconazole and fluorocytosine against C. albicans, C. tropicalis and A. niger. 19a-e (JQNNs) were generally showed relatively potent antifungal activities compared to fluconazole and fluorocytosine against C. krusei. Also, (15g) (JQES7) and (19b) (JQNN2) showed potent antifungal activities compared to fluconazole and fluorocytosine against all 6 fungi. But 10a-d (JQMSs) and 24a-j (COSs) did not show antifungal activity against all tested pathogenic fungi.;가. 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 항진균 작용 및 생리활성을 검색하였다. 1,3,4-Trisubstituted pyazole 유도체들은 두 종류의 acetophenone (4-Br, I)에 phenylhydrazine 또는 2-hydrazinopyridin을 반응하여 1-phenyl-2-(1-phenyl -ethylidene)hydrazines 2a-b와 1-(1-phenylethylidene)-2-(pyridin-2-yl) hydrazines 2c-d를 합성한 후 Vilsmeier-Haack complex (POCl₃/DMF)를 이용하여 네 종류의 1,3-disubstituted-1H-pyrazole-4-carbaldehyde 3a-d (JEAs)를 합성하였다. 각각의 JEAs를 LiAlH_(4)로 환원하여 (1,3-disubstituted-1H-pyrazol-4-yl) methanol 4a-d (JERs) 유도체 4개를 얻었다. 각각의 1,3-disubstituted-1H-pyrazole-4-carbaldehyde 3a-d (JEAs)에 당량의 2-amino-benzenethiol, 2-amino-4-chloro-benzenethiol 또는 benzene-1,2- diamine, 4,5-di-chloro-benzene-1,2-diamine를 반응하여 2-(1,3-disubstituted- 1H-pyrazol-4-yl)-1H-benzoimidazole, 2-(1,3-disubstituted-1H-pyrazol-4-yl) benzothiazole 5a-l (JEs) 유도체 12개를 얻었다. 마지막으로 각각의 1,3-disubstituted-1H-pyrazole-4-carbaldehydes 3a-d (JEAs)에 당량의 benzoic hydrazide를 넣어 [(1,3-disubstituted-1H-pyrazol-4-yl) methylene]-benzohydrazide 6a-d (JEBHs) 유도체 4개를 합성한 후 acetic anhydride로 cyclization하여 1-[2-(1,3-disubstituted-1H-pyrazol-4-yl)-5-phenyl-[1,3,4]-oxadiazol-3-yl]-ethanone 7a-d (JECYs) 유도체 2개를 얻었다. 나. Cinnoline계 유도체 화합물의 합성과 그 항진균 작용의 검색 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 JQMCYs, JQECY 유도체 5개를 합성하여 그 항진균 작용을 검색하였다. 모핵 Methyl 2-(2-chloro-1,4-dihydro-1,4-dioxonaphthalen-3-yl)-2-cyano-acetate (9) (JQM)와 ethyl 2-(2-chloro-1,4-dihydro-1,4-dioxonaphthalen-3-yl)-2-cyanoacetate (14) (JQE)에 당량의 arylthiols와 반응하여 각각 2-[2-arylthio-1,4-dihydro-1,4-dioxonaphthalen-3-yl]-2-cyanoacetate 10a-d (JQMSs) 유도체 4개, ethyl 2-(2-arylthio-1,4-dihydro-1,4-dioxo-naphthalen-3-yl)-2-cyanoacetate 15a-f (JQESs) 유도체 7개, 그리고 benzo[h]cinnoline 계의 새로운 모핵 6-hydroxy-cinnoline 유도체인 methyl 5-arylthio-3-amino-6-hydroxy-benzo[h]cinnoline-4-carboxylate 12a-c(JQMSCYs) 유도체 3개를 합성하였다. 다. 항진균성 benzo[f]indole-4,9-dione 유도체 화합물의 설계 합성 항암, 정균작용이 우수하다고 예상되는 heterocyclic indoledione인 benzo[f]indole-4,9-dione 유도체 중 새로운 모핵인 2-(3-chloro-4-hydroxy-1-oxonaphthalen-2(1H)-ylidene)malono-nitrile (17) (JQN)을 합성한 후, 여기에 당량의 benzenethiol을 반응시켜 (18) JQNS 유도체 1개를, 4당량의 arylamines 또는 aliphatic amines와 반응 시켜 분자내 고리화 반응을 시켜 다양한 indole 유도체인 2-amino-4,9-dihydro-4,9-dioxo-1-substituted-1H-benzo[f]indole-3-carbonitrile 19a-e (JQNNs) 유도체 5개를 합성하였다. 라. 항진균성 6-methyl-2H-chromene-2,5,8-trione 유도체 화합물의 설계 합성 항진균 작용이 있는 heterocyclic quinones 물질의 생리활성을 연구하기 위하여, 새로운 quinone 모핵인 6-methyl-2H-chromene-2,5,8-trione (23) COU을 합성하여 1/4 당량에 해당하는 arylthiols을 반응시켜 6-methyl-7-arylthio-2H-chromene-2,5,8-triones 24a-j (COSs) 유도체 10개를 합성하여 항진균 작용을 검색하였다. 합성한 유도체 (JQM, JQMS, JQMCYs, JQE, JQECY, JQESs, JQE7, JQNN, JQNS, COSs)들에 대해서 6가지 병원균주인 C. albicans, C. tropicalis, C. krusei, C. neoformans, A. niger, A. flvus에 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. 2-[2-(2-Hydroxyethylthio)-1,4-dihydro-1,4-dioxo-naphthalen-3-yl]-2-cyanoacetate 15g (JQES7)은 6가지 균주 모두에 대해서 대조약물인 fluconazole보다 항진균 효과가 좋았고, 특히, C. krusei 균주에 대해 대조약물인 fluconazole, fluorocytosine 보다 MIC 농도 3.2로 뛰어난 항진균 효과를 보였지만, JQES7을 제외한 그 외 ethyl 2-[2-arylthio-1,4-dihydro-1,4-dioxo-naphthalen-3-yl]-2-cyanoacetate1 5a-f (JQESs) 유도체들은 대체로 효과가 좋지 않았다. 또한, 6-hydroxycinnoline 유도체들에 대해서도 항진균 작용을 검색해 본 결과, methyl 5-(p-tolylthio)-3-amino-6-hydroxybenzo[h]cinnoline-4-carboxylate 12c (JQMSCY4)는 C. albicans, C. tropicalis, A. niger 균주에 대해서 MIC 농도 0.8과 1.6으로 대조약물들 보다 뛰어난 항진균 효과를 나타내었고, C. krusei, A. flavus에 대해서는 대조약물들과 효과가 비슷하거나 좋았지만, JQMSCY4를 제외한 다른 cinnoline 유도체들 methyl 3-amino-5-chloro-6-hydroxybenzo[h]cinnoline-4-carboxylate (11) (JQMSCY1), 5-arylthio-3-amino-6-hydroxybenzo[h]cinnoline-4-carboxylate 12a-c (JQMSCY2-4), ethyl 3-amino-5-chloro-4a,10b-dihydro-6-hydroxybenzo[h]cinnoline-4-carbo-xylate (16) (JQECY)은 효과가 좋지 않았다. 2-[1,4-Dihydro-1,4-dioxo-2-(phenylthio)naphthalen-3-yl]malononitrile (18) (JQNS)와 대부분의 2-amino-4,9-dihydro-4,9-dioxo-1-substituted-1H-benzo[f]indole-3-carbonitrile 19a-e (JQNNs)유도체들은 C. krusei 균주에 대해서 MIC 농도 6.3 이하로 대조약물인 fluorocytosine 보다 훨씬 좋은 항진균 효과를 보이고, fluconazole과는 비슷하거나 더욱 강력한 효과를 보였다. 특히, 2-amino-4,9-dihydro-1-methyl-4,9-dioxo-1H-benzo[f]indole-3-carbonitrile 19a (JQNN1), 2-amino-1-ethyl-4,9-dihydro-4,9-dioxo-1H-benzo[f]indole-3-carbonitrile 19b (JQNN2)는 6가지 균주에 대해 fluconazole 보다 효과가 우수했다. 한편, 2-[2-arylthio-1,4-dihydro-1,4-dioxonaphthalen-3-yl]-2-cyanoace-tate 10a-d (JQMSs)와 6-methyl-7-arylthio-2H-chromene-2,5,8-triones 24a-j (COSs) 유도체들은 모든 균주에서 대체로 효과가 좋지 않은 것으로 나타났다.