6,7-DISUBSTITUTED-5,8-QUINOLINEDIONE DERIVATIVES AS AN ANTIFUNGAL AGENT

Abstract: Novel 6,7-disubstituted-5,8-quinolinedione derivatives of general formula (I) or the process for the preparation thereof, wherein R1 is C1-C20 alkylmercapto or phenylamino substituted by up to 3 groups selected from halogen, aceto; R2 is halogen, thiocyano or C1-C20 alkylmercapto, are useful as an antifungal agent.

Description: 6,7-DISUBSTITUTED-5,8-QUINOLINEDIONE DERIVATIVES AS AN ANTIFUNGAL AGENT BACKGROUND OF THE INVENTION Field of the Invention The present invention relates to 6,7-disubstituted-5,8-quinolindion derivatives as an antifungal agent expressed in the following formula I which has excellent antifungal activity against funges infecting human and animals, l ϋ wherein Ri represents phenylamino group substituted 1 to 3 with a substituent selected from the group consisting of a halogen atom and an aceto group, or C1-C20 alkylmercapto group; and 1.1 R 2 represents thiocyano group; or G-C20 alkylmercapto group. Description of the Prior Art The recent increase of fungal infections has generated a renewed interest in antifungal drugs, including development of new antifungal agents. 20 We focused on developing 6,7-disubstituted-5,8-quinolinediones with noble mode of antifungal activities. As antimetabolites of coenzyme Q, the 6- (substituted)-7-chloro-5,8-quinolinediones inhibit mitochondrial Co-Q dependent succinoxidase and electron transport in Saccaromyces cerevisiπe , that may be correlated with antifungal activity (Arch. Biochem. Biophi/s., 191, 2,1 306-315, 1978). The 5,8-quinolindiones, as antimetabolites of coenzvme Q, inhibit cvtochrome be complex due to blockade of mitochondrial electron transport in fungi (/. Med. Client., 16, 206-209, 1973). 6/7-Substiruted-5,8- quinolinedione derivatives have also been reported fungicide in German Patent Publication No. 4, 208, 874 (1993), but no antifungal activity against human pathogenic fungi was not tested or mentioned. SUMMARY OF THE INVENTION The present invention has been completed by synthesizing 6,7- disubstituted-5,8-quinolinedion derivatives and confirming the antifungal activity thereof. Therefore, an object of the present invention is to provide 6,7- disubstituted-5,8-quinolinedion derivatives expressed in the formula 1, including its preparing method and use as an antifungal agent. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT The present invention relates to 6,7-disubstituted-5,8-quinolinedion derivatives expressed in the formula 1, wherein Ri represents phenylamino group substituted 1 to 3 with a substituent selected from the group consisting of a halogen atom and aceto group or Cj- C 2 o alkylmercapto group; and R 2 represents thiocyano group or C1-C20 alkylmercapto group. The present invention is described in more detail as set forth hereunder. Among 6,7-disubstituted-5,8-quinolinedion derivatives expressed in the formula 1, preferable compounds are the following formulas la, lb, l c and Id, wherein R ά , h and R are same or different and a hydrogen atom, a halogen atom or aceto group; and X is a halogen atom; wherein R d , Rb and R c are the same as defined for the compound la; 0 wherein R<„ Rb and R c are the same as defined for the compound la; and wherein Rd is C1-C20 alkyl group; and X is a halogen atom. ,1 The examples of 6,7-disubstituted-5,8-quinolindion derivatives of formula 1 are as follows; 6-[(N-4-chlorophenyl)amino]-7-bromo-5,8-quinolindion (compound 1), 6-[(N-3,5-dichlorophenyl)amino]-7-chloro-5,8-quinolindion (compound 2), 6-[(N-4-bromophenyl)amino]-7-bromo-5,8-quinolindion (compound 3), 6-[(N-4-acetophenyl)amino]-7-chloro-5,8-quinolindion (compound 4), 6-[(N-2,3,4-trifulorophenyl)amino]-7-methylmercapto-5,8-quinolindion (compound 5), 6-[(N-4-chlorophenyl)amino]-7-methylmercapto-5,8-quinolindion (compound 6), 6-[(N-4-iodophenyl)amino]-7-methylmercapto-5,8-quinolindion (compound 7), 6-[(N-4-iodophenyl)amino]-7-thiocyano-5,8-quinolindion (compound 8), 6-[(N-4-bromophenyl)amino]-7-thiocyano-5,8-quinolindion (compound 9), 6-[(N-4-chlorophenyl)amino]-7-thiocyano-5,8-quinolindion (compound 10), 6-[(N-4-acetophenyl)amino]-7-thiocyano-5,8-quinolindion (compound 11 ), 6-[(N-3,5-difulorophenyl)amino]-7-thiocyano-5,8-quinolindion (compound 12), 6-//-ethylmercapto-7-chloro-5,8-quinolindion (compound 13), 6-//-propylmercapto-7-chloro-5,8-quinolindion (compound 14), 6-/;-butylmercapto-7-chloro-5,8-quinolindion (compound 15), 6-/;-pentylmercapto-7-chloro-5,8-quinolindion (compound 16), 6-//-hecxylmercapto-7-chloro-5,8-quinolindion (compound 17), 6-/;-heptylmercapto-7-chloro-5,8-quinolindion (compound 18), 6-//-octylmercapto-7-chloro-5,8-quinolindion (compound 19), 6-//-nonylmercapto-7-chloro-5,8-quinolindion (compound 20). A method of preparing 6,7-disubstituted-5,8-quinolindion derivatives of formula 1 is shown in the following scheme 1, wherein R d/ Rb, R t , Rd and X are the same as defined above. A method of preparing 6,7-dihalo-5,8-quinolindione as starting material expressed in the following formula 2 in the present invention has been disclosed in /. am. Chew. Soc, 82, 1155 (1960) and Ann. tier Chein., 624, 108-119 (1959). It is prepared from 8-hydroxyquinoline through 5-nitroso-8- hydroxyquinoline and 5-amino-8-hydroxyquinoline chloride as intermediates. The detailed preparing method in scheme 1 is described hereunder. The compound of formula la is prepared by condensation reaction of 6,7-dihalo-5,8-quinolindione of formula 2 and an arylamine of formula 3. A solvent used in this reaction is C1-C3 lower alcohol such as methanol, ethanol and isopropanol, acetic acid, dimethylsulfoxide(DMSO) or dioxane, preferably ethanol. The reaction is carried at the temperature of from room temperature to 100 ° C for 2-6 hours, preferably at reflux temperature for 3-5 hours. The compound of formula lb is prepared by reacting compound la dissolved in an alcohol solvent with sodium sulfite(Na 2 S) at room temperature for 2-5 hours, preferably 2-3 hours. ft The compound of formula lc is prepared by reacting compound la with ammonium thiocyanate(NH4SCN) in acetone. Acetone as a solvent can be replaced to an alcohol such as methanol and ethanol, preferably ethanol. The compound of formula Id is prepared by reacting 6,7-dihalo-5,8- 1 quinolindione of formula 2 with alkylmercaptane(Rd-SH) at the same reaction condition of the preparing method of compound la. These compounds of formulas la, lb, lc and Id are isolated and purified by conventional procedures such as recrystallization and chromatography. 10 The following examples are intended to be illustrative of the present invention and should not be construed as limiting the scope of this invention defined by the appended claims. Example 1 : 5-nitroso-8-hydroxyquinoline hydrochloride 1.1 To a mixture of 8-hydroxyquinoline (58 g, 0.4 mol) in distilled water, concentrated hydrochloric acid (75 in?) and ice (200 g) was added NaN0 2 (30 g) in water (100 ml) at 0-4 ° C during 1 hr. The reaction mixture was kept at 0 ° C overnight and then filtered by washing with cold water to give 5-nitroso- 8-hydroxyquinoline hydrochloide (95%). 20 Example 2 : 5-amino-8-hydroxyquinoline dihydrochloride 5-nitroso-8-hydroxyquinoline chloride (40 g) was added to a mixture of water (160m?) and 5N-NaOH (260 mi) and heated to 40 TJ . Na 2 S 2 0 4 (95 g) was added to the reaction mixture with increasing temperature to 75-80 ° C . The reaction mixture was cooled to 50 ° C and 12N-HC1 (250 ml) was added to 21 it. Then the reaction mixture was cooled to 0 ° C and filtered to give 5-amino- 8-hydroxyquinoline dihydrochloride (34 g, 69%) Example 3 : 6,7-dichloro-5,8-quinolindion of formula 2 After addition of 5-amino-8-hydroxyquinoline dihydrochloride (9 g) to hydrochloric acid (81 g), the reaction mixture was heated to 60 °C and NaClO^ (4.5 g) was added. The reaction mixture was stirred for 30 min at 50-60 °C, 1 filtered and recrystallized twice with buthanol to give yellow precipitate of 6,7-dichloro-5,8-quinolindion (90%) m. p. : 221-222 ° C Example 4 : 6,7-dibromo-5,8-quinolindione of formula 2 0 After addition of 5-amino-8-hydroxyquinoline dihydrochloride (9 g) to hydrobromic acid (81 g), the reaction mixture was heated to 60 ° C and NaBrOi (7.5 g) was added. The reaction mixture was stirred for 30 min at 50-60 °C, filtered and recrystallized twice with buthanol to give yellow precipitate of 6,7-dibromo-5,8-quinolindion (67% ) ,1 m. p. : 243-245 ° C Example 5 : 6-[(N-4-chlorophenyl)amino]-7-bromo-5,8-quinolindione of formula 1 CeCl 3 (0.1 mmol) as a catalyst and 4 equivalents of 4-chloroaniline (10 0 mmol) were added to 6,7-dibromo-5,8-quinolindion (10 mmol) dissolved in ethanol (100 ml). The reaction mixture was heated at reflux for 8 hrs. After reading the completion of reaction by TLC, it was cooled, filtered, and recrvstallized with ethanol to give redish powder of 6-[(N-4- chlorophenyl)ammo]-7-bromo-5,8-qumotidione. ,1 Example 6: 6-[(N-4-chlorophenyl)amino]-7-methylmercapto-5,8- quinolidione of formula 6 Dimethylsulfate (12 mmol) was added to 6-[(N-4- chlorophenyl)amino]-7-bromo-5,8-quinolidione (10 mmol) dissolved in ethanol (100 ml). The reaction mixture was stirred for 2 hrs at room temperature The reaction mixture was cooled, filtered, and recrvstallized with ethanol to give 6-[(N-4-chlorophenyl)amino]-7-methylmercapto-5,8- quinolidione. Example 7 : 6-[(N-4-chlorophenyl)amino]-7-thiocyano-5,8-quinolidione of formula 10 1 equivalent of ammonium thiocyanate(NH4SCN) was added to 6-[(N- 4-chlorophenyl)amino]-7-bromo-5,8-quinolidione (1 g). The reaction mixture was heated at reflux for 3 hrs. After reading the completion of reaction by TLC, it was cooled, filtered, and recrystallized with ethanol to give 6-[(N-4- chlorophenyl)amino]-7-thiocyano-5,8-quinolidione. l Example 8 : 6-ethylcercapto-7-chloro-5,8-quinolindione of formula 13 CeCl 3 (0.1 mmol) as a catalyst and 1 equivalent of sodium sulfide(Na2S) were added to 6,7-dichloro-5,8-quinolindion (1 g) dissolved in ethanol (100 ml). The reaction mixture was stirred for 30 min at room 20 temperature. Λ equivalent of ethyl mercaptane was added to the reaction mixture and heated at reflux for 1 hr. The reaction mixture was cooled and filtered to give 6-ethylmercapto-7-chloro-5,8-quinolidione. Physical properties of the compounds of formula 1 prepared by the 21 above procedure are summarized in table la and table lb. Table la Table ib The compounds of 6,7-disubstituted-5,8-quinolindione derivatives of formula 1 and its pharmaceutically acceptable salts have excellent antifungal activity. A pharmaceutical composition containing the compounds as an effective ingredient is disclosed in the present invention. Said pharmaceutically acceptable salts include hydrochloric acid salt, sulfuric acid salt and bisulfite salt. A pharmaceutical composition comprises a compound of formula 1, pharmaceutically acceptable carriers, fillers, and other additives. The carrier(s) must be 'acceptable' in the sense of being compatible with other .1 ingredients of the formulation and not deleterious to the recipient thereof. The compositions of the invention include those in a foam especiallv formulated for a)oral administration such as tablets, capsules, troches, solutions, emulsions and suspensions, b)parenteral administration bv injection presented in such forms as suspension, solutions, emulsions and 10 powder for constitution with a suitable vehicle, e.g. sterile water, before use, and c)topical administration in the form of ointments, creams, gels, or lotions. Pharmaceutically acceptable carriers in use for suitable formulations are: binding agents, lubricants, diluents, solubilizers, disintegrants, stabilizers, emulsifying agents, colorants and aromatics for oral administration; 1.1 preservatives, solubilizers, and stabilizers for parenteral administration; and bases, diluents, lubricants, thickeners, and preservatives for topical administration. These pharmaceutically acceptable formulations are administered orally, parenterally by intravenous, subcutaneous and intraperitoneal injection/ infusion, or topically. In oral administration, 20 antacids can be combined in a pharmaceutical composition to prevent from degradation by gastric acid and tablets may be coated to give enteric-coated tablet. The dose varies depending on age, body weight, sex, symptom and the like of patients, and types of administration but it is usually 0.01-200 2.1 mg/kg/day, which is administered in several doses a day, preferably in a single dose or several doses. Examples for formulation comprising 6,7-disubstituted-5,8- quinolindion derivatives of formula 1 and its pharmaceutically acceptable salts as an effective ingredient are described but it should be noted that the present invention is not limited to these exemplifications. 1 Formulation Example 1 : Capsule for oral administration 100 mg of an effective compound, 10 mg of anhydrous silicic acid, 190 mg of microcrystalline cellulose, 5 mg of magnesium stearate, 60 mg of starch and sodium gluconate and 135 mg of anhydrous calcium hydrogenphosphate were mixed and stirred. The mixture was put into hard gelatin capsules to l' i give a capsule drug. Formulation 2 : Injection for parental administration 100 mg of an effective compound, 180 mg of mannitol, 26 mg of sodium hydrogenphosphate were dissolved in 2,974 mg of sterile water to 1.1 give an injection solution. Formulation 3 : Ointment for topical administration 2 g of an effective compound, 7 g of lanoline tablets and effective amount of white ointment to give 10 g of total weight. 20 Antifungal activities of the compounds of formula 1 according to the present invention are determined by the following Experimental Examples. Experimental Example 1 : Determination of Minimum Inhibitory Concentration(MIC) Values 2.1 The minimum inhibitory concentration (MIC), which indicates the lowest concentration of the active ingredient in the growth medium, was determined by solid medium dilution method. Pathogens strains used are Candida albicans ATCC10231, Candida glabrata ATCC2001, Candida tropicπlis ATCC28775, Candida parasilosis ATCC22019, Cryptocoaiss neofornians KCTC7223. 4 mg of a test compound was dissolved in 2 m l of .1 dimethylfulfoxide(DMSO) to produce solutions having a concentration of 2 mg/ml for use and serial two-fold dilution was prepared. The test medium was Sabouraud's agar. 50 ? aliquot of serial two-fold dilution was added to 1 ml of agar medium in 10 x 1.2 cm petri dishes to be 100, 50, 25, 12.5 6.3, 1.6, and 0.8 /ig/ml of test compound concentration and the slant was prepared by H) tilting the petri dishes. The MIC value without containing a test compound was determined in order to test an effect of a solvent to an antifungal activity. Hvphae with a diameter of 2 mm were inoculated with 2 x 10 5 CFU. After the medium was cultured for 3 davs at 30 ° C, the MIC values were determined. Control drugs were fluconazole and ketoconazole. The datas are shown in 11 table 2. 0 5 Table 2 As shown in table 2, the compounds of the present invention show excellent inhibitory activity against Candida albicans, C. glabrata, C. tropicalis, C. parapsilosis, Cryptococuss neoformans and the like and their activities are superior or similar to those of the control drugs, fluconazole and ketoconazole. Especially, the growth of fungus was completely inhibited against 5 types of strains when the concentration of the compound was 50 /ig/ml. Experimental Example 2 : C. albicans (1 x 10 7 ) isolated was inoculated to Sabouraud's agar, suspended in a physiological saline solution, and injected into tail vein of a rat. Strains to kill a rat within 48 hours were selected. C. albicans (2 x 10 6 which is concentration to kill a rat within 48 hours) suspended in a physiological saline solution was injected and infected into tail veins of 125 rats. Each 0 concentration (10, 2, 0.5, 0.1, 0.025 mg/kg) of the compounds of table 3 was administered to each 10 rats. After infecting with Candida strain, each compound was administered to each 10 rats. Fluconazol as a control drug was also administered with 10, 2, 0.5 and 0.1 mg/kg of concentrations. A physiological saline solution containing 0.2 % tween 20 was administered as a ,1 control group. The datas are shown in table 3. Table 3 According to the table 3, while ED50 of a control drug, fluconazole, was 0 6.00 mg/kg of the concentration to a rat infected with C. albicans, each ED50 of the compounds 1, 2, 3, and 4 was 0.06, 0.07, 0.06 and 0.04 mg/kg of the concentration. Experimental Example : Survival prolongation effect of a rat infected with C. albicans 2 x 10 4 /0.1 ml of C. albicans suspended in physiological saline was injected into 49 rats via tail vein to infect and divided to 4 groups of 7 rats. The concentration of effective dose (ED50) was injected intraperitoneally once daily for 14 days after 4 days of infection. In other words, 0.06, 0.07, 0.06 and 0.04 mg/kg of the test compounds 1, 2, 3, and 4 and 6.00 mg/kg of the control drug, ketoconazole were injected to determine the survival rate. The results were shown in Figs. 1 through 4. According to Figs. 1 through 4, the compounds of the present invention show similar inhibitory activity at lower concentration than that of ketoconazole which is the control drug. Therefore, 6,7-disubstituted-5,8- quinolindione derivatives of formula 1 have excellent inhibitory activity against various fungus. Based on foregoing, the compounds of formula 1 of the present invention are useful for the treatment of a variety of fungal infections in human and animals.

Claims: CLAIMS What is claimed is : 1. 6,7-disubstituted-5,8-quinolindione derivatives represented in the formula 1, wherein Rι represents a phenylamino group substituted 1 to 3 with a substituent selected from the group consisting of a halogen atom and an aceto group; or Cι-C 2 o alkylmercapto group; and R 2 represents a thiocyano group; or C1-C20 alkylmercapto group. 2. The 6,7-disubstituted-5,8-quinolindione derivatives according to claim 1, wherein said compounds are represented in formula la, wherein R d , Rb and R c are same or different and a hydrogen atom, a halogen atom or aceto group; and X is a halogen atom. 3. The 6,7-disubstituted-5,8-quinolindione derivatives according to claim 1, wherein said compounds are represented in formula lb, wherein R ( „ Rb and R t are the same as defined for the compound la. 4. The 6,7-disubstituted-5,8-quinolindione derivatives according to claim I, wherein said compounds are represented in formula lc, wherein Ra, Rb and R c are the same as defined for the compound la. 5. The 6,7-disubstituted-5,8-quinolindione derivatives according to claim 1, 10 wherein said compounds are represented in formula Id, wherein Rd is G-C20 alkyl group; and X is a halogen atom. 1.1 6. The 6,7-disubstituted-5,8-quinolindione derivatives according to claim 1, wherein said compound is selected from the group consisting of ; 6-[(N-4-chlorophenyl)amino]-7-bromo-5,8-quinolindion, 6-[(N-3,5-dichlorophenyl)amino]-7-chloro-5,8-quinolindion, 6-[(N-4-bromophenyl)amino]-7-bromo-5,8-quinolindion, 20 6-[(N-4-acetophenyl)amino]-7-chloro-5,8-quinolindion, 6-[(N-2,3,4-trifulorophenyl)amino]-7-methylmercapto-5,8-quinolindion, 6-[(N-4-chlorophenyl)amino]-7-methylmercapto-5,8-quinolindion , 6-[(N-4-iodophenyl)amino]-7-methylmercapto-5,8-quinolindion, 6-[(N-4-iodophenyl)amino]-7-thiocyano-5,8-quinolindion, 6-[(N-4-bromophenyl)amino]-7-thiocyano-5,8-quinolindion, ,1 6-[(N-4-chlorophenyl)amino]-7-thiocyano-5,8-quinolindion, 6-[(N-4-acetophenyl)amino]-7-thiocyano-5,8-quinolindion, 6-[(N-3,5-difulorophenyl)amino]-7-thiocyano-5,8-quinolindion, 6-//-ethylmercapto-7-chloro-5,8-quinolindion, 6-;/-propylmercapto-7-chloro-5,8-quinolindion, 6-/7-butylmercapto-7-chloro-5,8-quinolindion, 6-/;-pentylmercapto-7-chloro-5,8-quinolindion, 6-/;-hecxyl ercapto-7-chloro-5,8-quinolindion, 6-//-heptylmercapto-7-chloro-5,8-quinolindion, 6-//-octylmercapto-7-chloro-5,8-quinolindion, and 6-//-nonylmercapto-7-chloro-5,8-quinolindion. 7. A process for preparing the compound of formula la by reacting 6,7- dihalo-5,8-quinolindion represented by formula 2 and an arylamine represented by formula 3, wherein R d , R and R c are same or different and a hydrogen atom, a halogen atom or an aceto group; and X is a halogen atom. 8. The process for preparing the compound of formula la according to claim 7, wherein the solvent is selected from the group consisting of alcohol, glacial acetic acid, dimethylsulfoxide and dioxane. 9. The process for preparing the compound of formula la according to claim 8, wherein said solvent is ethanol. 10. The process for preparing the compound of formula la according to claim 7, wherein said reaction is performed at room temperature to 100 °C . 11. The process for preparing the compound of formula la according to claim 7, wherein said reaction is preformed for 2 to 6 hours. 12. A process for preparing the compound of formula lb by reacting the compound of formula la with sodium sulfide and dimethylsulfate ((CH 3 ) 2 S0 2 ), wherein Ra, Rb and R c are same or different and a hydrogen atom, a halogen atom or an aceto group; and X is a halogen atom. 13. A process for preparing the compound of formula lc by reacting the compound of formula la with ammonium thiocyanate(NH 4 SCN), wherein R a , Rb and R c are same or different and a hydrogen atom, a halogen atom or an aceto group; and X is a halogen atom. 14. The process for preparing the compound of formula lc according to claim 13, wherein said reaction is performed in acetone by heating at reflux. 15. The process for preparing the compound of formula lc according to claim 13, wherein said reaction is performed for 2 to 6 hours. 16. A process for preparing the compound of formula Id by reacting 6,7- dihalo-5,8-quinolindione of formula 2 with alkylmercaptane(Rd-SH), wherein Rd is C1-C20 alkyl group; and X is a halogen atom. 17. An antifungal agent comprising 6,7-disubstituted-5,8-quinolindione derivatives of formula 1 or its pharmaceutically acceptable salts, wherein Ri represents a phenylamino group substituted 1 to 3 with a substituent selected from the group consisting of a halogen atom and an aceto group; or C1-C20 alkylmercapto group; and R 2 represents a thiocyano group; or C1-C20 alkylmercapto group.

Inventors: RYU CHUNG KYU, RYU, CHUNG, KYU
Applicants: IL JIN COPPER FOIL CORP, RYU CHUNG KYU, IL JIN COPPER FOIL CORP., RYU, CHUNG, KYU
Application year: 2000
Main classification: C07D215
IPC codes: A61K31, A61P31, C07D215