QUINOLONECARBOXYLIC ACID DERIVATIVES

Abstract: Quinolonecarboxylic acid derivatives of the following formula, wherein R, R¹ and R² are each independently hydrogen atom or lower alkyl group and Y is hydrogen atom or halogen atom; the hydrates and pharmaceutically acceptable salts thereof are useful as antibacterial agents.

Description: Quinolonecarboxylic acid derivatives Priority: November 13, 1984, Japan, Sho 59-239124 (P) Detailed description of the invention: This invention is concerned with certain novel useful quinolonecarboxylic acid derivatives, having antibacterial activities, with a process for their preparation, and with compositions containing them. This invention provides compounds of the formula (I) EMI1.1 wherein R, R1 and R2 are each independently hydrogen atom or lower alkyl group and Y is hydrogen atom or halogen atom; the hydrates and pharmaceutically acceptable salts thereof. Since nalidixic acid which has been employed for treatment of urinary tract infections by gram-negative bacteria, was introduced in 1963, intensive work has been carried out on the further development of quinolonecarboxylic acid analogue. Thus, recently a remarkable antibacterial activity against not only gram-negative but also gram-positive bacteria occurs for some compounds (e.g. norfloxacin). However their activity against gram-positive bacteria is fairly less than that against gram-negative bacteria. Just recently, the drugs which have relatively strong activity against gram-positive bacteria has been developed, but shown to possess weaker activity against gram-negative bacteria than that of the prior compounds (e.g. norfloxacin, ciprofloxacin). As a result of the investigation, the present inventors have now unexpectedly found that new derivatives of quinolonecarboxylic acid represented by the formula (I) have excitingly potential activity against gram-positive bacteria without decrease of activity against gram-negative bacteria in comparison with that of any prior analogue and therefore are superior to commercial preparations and investigational drugs in the in vitro and in vivo antibacterial activity against -both gram-negative and gram-positive bacteria. Especially, the 8-chloro and 8-bromo compounds according to the present invention are valuable in human and veterinary medicine because of their greater activity and broader spectrum against both aerobic and anaerobic bacteria. While the compounds of the present invention show such -. strong activities against bacteria, their toxicity against mammalian cells is very weak. The present compounds are well absorbed and distributed into the tissue when administered orally to animals. The present compounds, therefore, are active at low doses against both gram-positive and gram-negative bacteria and thus constitute valuable agents for the treatment of infectious human, animal or plant diseases. The compounds of the formula (I) are synthesized by reacting a compound of the formula (11), EMI3.1 wherein X is halogen atom, R and Y are the same as defined above, with a compound of the formula (III), EMI3.2 wherein R1 and R2 are the same as defined above. The reaction is preferably carried out by heating the two reactants in a solvent such as water, alcohols, acetonitrile, dimethylformamide (DMF), dimethyl sulfoxide (DMSO), hexamethylphosphoric triamide, pyridine, picoline and the like or in absence of the solvent with, if necessary, an acid acceptor such as an inorganic or organic acceptor, e.g., alkali metal carbonate such as potassium carbonate or tert-amine, such as triethylamine, diazabicyclo base at room temperature to 200'C, preferably room temperature to 160'C, more preferably room temperature to 1200C for 1 to several hours. It is desirable that a slight excess (1 to 5 moles) of the secondary amine of the formula (III) is used per mole of the compound of the formula (II) and a solvent is used such that the mixture remains homogeneous after dissolution of the compound of the formula (II) (2 to l0-fold volume per volume of the compound of the formula (III)). When R in the formula (II) is lower alkyl group, the reaction product (carboxylic ester) is hydrolyzed to the corresponding carboxylic acid by the usual manner. The hydrolysis is carried out by treating the compound of formula (I2R is lower alkyl group) with alkali metal hydroxide solution such as sodium hydroxide, potassium hydroxide, or mineral acid such as hydrochloric acid, sulfuric acid in water, aqueous alcohols or an appropriate solvent. Furthermore, the compounds of the formulaI) I) can be converted, if desired, to the pharmaceutically acceptable salts by treatment with acid or alkali. The acid may be organic or inorganic acids such as, for example, hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid,-acetic acid, oxalic acid and lactic acid. The alkali salts may be, for example. sodium, potassium, magnesium, calcium, aluminum, cerium, chromium, cobalt, copper, iron, zinc, platinum and silver salts. The compound of the formula (I), the hydrates and salts thereof may be used as medicines in the conventional form of pharmaceutical preparations, which may be, for example, tablets; capsules, powder, ointments, supositories, injections or eye drops, suitable for peroral, parenteral, enteral or local administration. The following examples will further illustrate the invention without, however, limiting it thereto. Example 1. 7-t3-Aminomethyl-l-pyrrolidinyl)-l-cyclopropyl-6,8- difluoro-1,4-dihydro-4-oxo-3-quinolinecarboxylic acid hydrochloride A mixutre of ethyl l-cyclopropyl-6,7,8-trifluoro-1,4- dihydro-4-oxo-3-quinolinecarboxylate (0.3 g), 1,8-diazabicyclo (5,4,0)-7-undecene (DBU, 0.15 g) and 3-aminomethylpyrrolidine (0.14 g) in acetonitrile (5 ml) was refluxed under stirring for 3.5 hours. After cooling, the mixture was poured into ice-water (50 ml) and extracted three times with chloroform (50 ml). The extracts were washed with water, dried over anhydrous sodium sulfate and concentrated to give the residue, ethyl 7-(3-amino methyl-l-pyrrolidinyl)-l-cyclopropyl-6.8-difluoro-4-oXo-3- quinolinecarboxylate. To the residue was added 1sodium hydroxide solution (6 ml) and the mixture was stirred at 100 C for an hour. After cooling, the mixture was neutralized by adding acetic-acid and concentrated. Purification of the residue by silica gel column chromatography (CHCl3 s MeOH t con.NH4 oH=1O : 3 : 3) gave the title compound (54 mg) as pale yellow crystals, mp 248-250"C, after recrystallization from acetone-HCl (1 %). Analysis (%) for C18H19F2N303.HCl.1/3 H2O; Calcd. (Found): C, 53.27 (53.40); H, 5.13 (4.98); N, 10.35 (10.35). Example 2. 1-Cyclopropyl-7-(3-ethylaminomethyl-1-pyrrolidinyl)- 6,8-difluoro-1,4,dihydro-4-oxo-3-quinolinecarboxylic acid hydrochloride A mixture of ethyl l-cyclopropyl-6,7,8-trifluoro-1,4- dihydro-4-oxo-3-quinolinecarboxylate (0.3 g), DBU (0.15 g) and 3 ethylaminornethylpyrrolidine (0.13 g) in acetonitrile (5 ml) was refluxed under stirring for 3.5 hours. After cooling, the reaction mixture was poured into ice-water (25 ml) and extracted with chloroform (50 ml). The extract was washed with water, dried over anhydrous sodium sulfate and concentrated. To the residue, ethyl cyclopropyl-7-(3-ethylaminomethyl-1-pyrrolidinyl) 6, 8-difluoro-4-oxo-3-quinolinecarboxylate, was added 1N-sodium hydroxide solution (6 ml) and the mixture was stirred at 70 to 80'C for an hour. After cooling, the mixture was neutralized by adding acetic acid and concentrated. The residue, after purified by silica gel column chromatography (CHC13 t MeOH s con.NH4OH=10 s 10 s 3), was dissolved in methanol containinhydrochloric acid. The solution was concentrated to dryness and the solid was recrystallized from methanol to give the title compound (0.11 g), mp 258-260 C. Analysis (%)for C20H23F2N3Oc.HCl; Calcd. (Found): C, 56.14 (55.75) H, 5.65 (5.57) N, 9.82 (9.81). Example 3. 3. 1-Cyclopropyl-6,7,8-trifluoro-1,4-dihydro-4-oXo-3- quinolinecarboxylic acid A mixture of ethyl 1-cyclopropyl-6,7,8-trifluoro-1,4dihydro-4-oxo-3-quinolinecarboxylate (3.0 g), acetic acid (20 ml), sulfuric acid (2.5 ml) and water (15 ml) was refluxed under stirring for an hour. After cooling, the reaction mixture was poured into ice-water. The precipitate was filtered, washed sufficently with water and dried in vacuum to give the title compound (2.59 g) as colorless needles, mp 231-232"C. Analysis (%) for C13H8F3NO3; Calcd. (Found): C, 55.13 (55.11); H, 2.85 (2.61); N, 4.95 (4.79). Example 4. 7-(3-Aminomethyl-l-pyrrolidinyl)-l-cyclopropyl-6,8 difluoro-1,4-dihydro-4-oxo-3-quinolinecarboxylic acid A mixture of l-cyclopropyl-6,7,8-trifluoro-1,4-dihydro-4- oxo-3-quinolinecarboxylic acid (1.2 g), DBU (0.15 g) and 3aminomethylpyrrolidine (0.45 g) in anhydrous acetonitrile (10 ml) was stirred under reflux for 1.5 hours and then at room temperature for 9.5 hours. The precipitate formed was filtered and recrystallized from dichloromethane-methanol (1:1). The title compound (0.88 g), mp 235-236-C, was obtained as colorless prisms. Analysis (%) for C18H19F2N303; Calcd. (Fond): C, 59.50 (59.45); H, 5.27 (5.17) N, 11.56 (11.53). Example 5. 1-Cyclopropyl-7-(3-ethylaminomethyl-1-pyrrolidinyl) 6, 8-difluoro-1 , 4-dihydro-4-oxo-3-quinolinecarboxylic acid hydrochloride. A mixture of l-cyclopropyl-6,7,8-trifluoro-1,4-dihydro-4- oxo-3-quinolinecarboxylic acid (1.2 g), DBU (0.65 g) and 3ethylaminomethylpyrrolidine (0.57 g) in anhydrous acetonitrile (10 ml) was stirred under reflux for 1.5 hours and then at room temperature for 9.5 hours. The precipitate formed was filtered and dissolved in methanol containing hydrochloric acid. The solution was concentrated and the residue was recrystallized from water-ethanol to give the title compound (0.7 g) as pale yellow prisms, mp 256-258.5-C. Analysis (%) for C20H23 F2N303.HCl; Calcd. (Found); C, 56.14 (56.04); H, 5.65 (5.64); N, 9,82 (9,74). Example 6. 7-(3-Aminomethyl-l-pyrrolidinyl)-l-cyclopropyl-6 fluoro-1 , 4-dihydro-4-oxo-3-quinolinecarboxylic acid A mixture of 7-chloro-1-cyclopropyl-6-fluoro-1, 4-dihydro-4oxo-3-quinolinecarboxylic acid (0.5 g) and 3-aminomethylpyrrolidine (0.55 g) in B-picoline (18 ml) was refluxed for 3 hours under stirring. After cooling, to the mixture was added concentrated aqueous ammonia. The resulting mixture was concentrated to give the residue, to which acetonitrile (50 ml) was added. The precipitate was filtered, washed successively with ethanol-ether (its) and ether and recrystallized from methanol to give the title compound (0.12 g) as pale yellow prisms, mp 241-246-C. Analysis (%) for C18H20FN303 Calcd. (Found): C, 62.60 (62.30); H, 5.84 (5.71); N, 12.17 (12.11). Example 7. 1-Cyclopropyl-7-(3-ethylaminomethyl-1-pyrrolidinyl) 6-fluoro-1,4-dihydro-4-oxo-3-quinolinecarboxylic acid hydrochloride A mixture of 7-chloro-1-cyclopropyl-6-fluoro-1,4-dihydro-4- oxo-3-quinolinecarboxylic acid (0.6 g) and 3-ethylaminomethylpyrrolidine (0.81 g) in B-picoline (15 ml) was refluxed for 2 hours under stirring. After cooling, concentrated aqueous ammonia (20 ml) was added to the reaction mixture. The mixture was concentrated and the residue was dissolved in a mixture of concentrated hydrochloric acid and methynol. Removal of the solvent left a solid which was recrystallized from watermethanol-ethyl acetate (1 s 1 : 1) to give the title compound (125 mg) as colorless prisms, mp 282-285iC (decompd.). Analysis (%)for C20H24FN3O3.HCl, Calcd. (Found)&num; C, 58.10 (58.18); H, 6.19 (6.38); N, 10.16 (10.20). Example 8. 1-Cyclopropyl-6,8-difluoro-1,4-dihydro-7-(3-methyl aminomethyl-l-pyrrolidinyl)-4-oxo-3-quinoline- carboxylic acid hydrochloride A mixture of l-cyclopropyl-6,7,8-trifluoro-1,4-dihydro-4- oxo-3-quinolinecarboxylic acid (0.6 g), 3-methylaminomethylpyrrolidine (0.28 g) and DBU (0.33 g) in acetonitrile (5 ml) was stirred under reflux for an hour and then at room temperature for 8 hours. The reaction mixture was treated by the same way as described in Example 5. The title compound was obtained as pale yellow prisms (0.46 g), mp 269-273-C (decompds) after recrystallization from methanol. Analysis (t) for C19H21F2N3O3.HCl, Calcd. (Found): C, 55.14 (54.91); H, 5.36 (5.31); N, 10.15 (10.10). Example 9. 1-Cyclopropyl-6-fluoro-1,4-dihydro-7-(3-methylamino methyl-1-pyrrolidinyl ) -4-oxo-3-quinolinecarboxylic acid hydrochloride A mixture of 7-chloro-1-cyclopropyl-6-fluoro-1,4-dihydro-4- oxo-3-quinolinecarboxylic acid (0.6 g) and 3-methylaminomethylpyrrolidine (0.75 g) in B-picoline (4 ml) was stirred under reflux for 50 minutes and then at room temperature for 3 hours. The reaction mixture was treated by the same way as shown in Example 7. The title compound was obtained as pale yellow prisms (290 mg), mp 265 C (decompd.) after recrystallization from water. Analysis (%) for C19H22FN3O3'HCl1/5 H2O, Calcd. (Found): C, 57.13 (57.04); H, 5.90 (5.83); N, 10.52 (10.36). Example 10. 1-Cyclopropyl-6,8-difluoro-1,4-dihydro-4-oxo-7-(3-n propylaminomethyl-1-pyrrolidinyl) -3-quinoline- carboxylic acid hydrochloride A solution of ethyl l-cyclopropyl-6,7,8-trifluoro-1,4- dihydro-4-oxo-3-quinolinecarboxylate (0.3 g) and 3-n-propylaminomethylpyrrolidine (0.55 g) in DMF (10 ml) was stirred at 80-90-C for 2 hours. After cooling, the mixture was poured into icewater, alkalized by adding aqueous potassium carbonate solution and extracted with chloroform. The chloroform extract was dried over anhydrous sodium sulfate and then concentrated. Then residue, ethyl l-cyclopropyl-6,8-difluoro-1,4-dihydro-4-oxo-7-(3- n-propylaminomethyl-l-pyrrolidinyl)-3-quinolinecarboxylate in aqueous 1N sodium hydroxide solution (10 ml) was heated to reflux for an hour and then neutralized by adding acetic acid to give a crystalline precipitate which was collected by filtration and recrystallized from ethanol-water-acetonitrile. The product was dissolved in methanol containing hydrochloric acid and the solution was concentrated. The resulting residue was recrystallized from methanol to give the title compound (47 mg) as fine brown crystals, mp 270-278-C (decompd.). Analysis (%) for C21H25F2N3O3.HCl.4/5 H20, Calcd. (Found): C, 55.28 (55.13); H, 6.10 (5.76)t N, 9.21 (9.13). Example 11. 1-Cylopropyl-6,8-difluoro-1,4-dihydro-7-(3-iso propylaminomethyl-l-pyrrolidinyl)-4-oxo-3-quinoline carboxylic acid hydrochloride A mixture of ethyl 1-cyclopropyl-6,7,8-trifluoro-1,4- dihydro-4-oxo-3-quinolinecarboxylate (0.15 g) and 3-isopropylaminomethylpyrrolidine (0.27 g) in DMF (10 ml) was stirred at 80 900C for 2 hours. After cooling, the mixture was diluted with ice-water, alkalized by adding aqueous potassium carbonate solution and extracted with chloroform. The chloroform extract was dried over anhydrous sodium sulfate and then concentrated. The resulting residue, ethyl 1-cyclopropyl-6,8-difluoro-1,4- dihydro-7- ( 3-isopropylaminomethyl-1-pyrrolidinyl ) -4-oxo-3- quinolinecarboxylate in aqueous lN-sodium hydroxide solution was heated to reflux for an hour. The mixture was neutralized by adding acetic acid and concentrated. The resisue was purified by silica gel column chromatography (CHCl3 s MeOH s con.NH4OH=10 10 s 3). The product was dissoluved in methanol containing hydrochloric acid. Removal of the solvent left the title compound which was recrystallized from methanol to give fine brown crystals (15 mg), mp 265-271C (decompd.). Analysis (8) fbr C21H25F2N303.HCl.1/2 H2O, Calcd. (Found): C, 55.94 (56.04); H, 6.04 (5.85); N, 9.32 (9.31). Example 12. 8-Chloro-l-cyclopropyl-6-fluoro-1,4-dihydro-7-(3 methylaminomethyl-l-pyrrolidinyl)-4-oxo-3-quinoline carboxylic acid hydrochloride A mixture of 8-chloro-1-cyclopropyl-6,7-difloro-1,4- dihydro-4-oxo-3-quinolinecarboxylic acid (0.4 g), 3-methylamino- methylpyrrolidine (0,17 g) and DBU (0.2 g) in acetonitrile (4 ml) was refluxed for 2 hours and allowed to stand overnight during which time a crystal separated. The crystal, free form of the title compound, was collected by filtration and weighted 0.5 g. The product was dissolved in a mixture of concentrated hydrochloric acid and methanol. Removal of the solvent left the title compound which was recrystallized from ethanol to give yellow prisms (0.3 g), mp 206-210'C (decompd.). Analysis (*) for C19H21ClFN3O3.HCl1/3 H20, Calcd. (Found): C, 52.30 (52.05); H, 5.24 (5.03); N, 9.63 (9.45). In this example, the starting material is also novel and it is synthesized by following process. Reference example 1. N-(3-Chloro-4-fiuorophenyl)acetamide To 3-chloro-4-fluoroaniline (100 g) was slowly added acetic anhydride (200 ml). After allowed to stand for 30 minutes, the reaction mixture was poured into water (1 litter). The resulting precipitate was collected by filtration and recrystallized from aqueous ethanol to give the title compound (119.4 g), mp 118 1l90C. Reference example 2. N-(3-Chloro-4-fluoro-6-nitrophenyl) acetamide To a solution of N-(3-chloro-4-fluorophenyl)acetamide (55 g) in concentrated sulfuric acid (165 ml) was added dropwise concentrated nitric acid (d 1.42, 154 ml) at 5-10'C during an hour with stirring in an ice-salt bath. After stirring for an hour at the same temperature, the reaction mixture was poured into ice water. The resulting precipitate was collected by filtration, suf ficiently washed with water and recrystallized from acetonitrile to give the title compound (48.9 g) as yellow needles, mp 114 1150C. Analysis (8) for C8H6Cl FN2O35 Calcd. (Found): C, 41.31 (41.48); H, 2.60 (2.52) N, 12.04 (12.13). Reference example 3. 3-Chloro-4-fluoro-6-nitroaniline A solution of N-( 3-chloro-4-fluoro-6-nitrophenyl)acetamide (30 g) in concentrated hydrochloric acid (50 ml) and ethanol (200 ml) was refluxed for 2.5 hours. To the reaction mixture was added ice water (300 ml) and the resulting precipitate was collected by filtration, washed with water and dried to give the title compound (24.9 g) as yellow needles, mp 149.5-150 C. Analysis (%) for C6H4ClFN202, Calcd. (Found): C, 37.82 (37.85); H, 2.11 (2.03); N, 14.70 (14.80). Reference example 4. 2,3-Dichloro-4-fluoro-6-nitroaniline Into a solution of 3-chloro-4-fluoro-6-nitroaniline (14.3 g) in acetic acid (150 ml) was bubbled chlorine gas at 18-20-C during 70 minutes. The reaction mixture was poured into ice water (300 ml) and the resulting precipitate was collected by filtration, washed with water and recrystallized from ethanol to give the title compound (14.33 g) as yellow needles, mp 161-C. Analysis (%) for C6H3C12FN202, Calcd. (Found) sC, 32.03 (32.17); H, 1.34 (1.26); N, 12.45 (12.65). Reference example 5. 2,3,4-Trichloro-5-fluoronitrobenzene To a mixture of anhydrous cupric chloride (13.58 g) and tert-butylnitrite (12.4 g) in anhydrous acetonitrile (100 ml) was added portionwise 2,3-dichloro-4-fluoro-6-nitroaniline (18.05 g at 60-62-C during 30 minutes. After stirring for 30 minutes at 60-65*C, the reaction mixture was poured into chilled 10% diluted hydrochloric acid (300 ml) and extradited with benzene. The organic layer was successively washed with diluted hydrochloric acid and water, dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by distillation to give the title compound(17.26 g), bp 137-142'C/27 mmHg. NMR (d in CDC13), 7.65 (d, J=7.5 Hz) Reference example 6. 3-Chloro-2,4,5-trifluoronitrobenzene To a suspension of potassium fluoride (64.9 g) in anhydrous dimethyl sulfoxide (230 ml) was added 2,3,4-trichloro-5-fluoronitrobenzene (54.4 g) at 140'C and stirred at the same temperature for 10 minutes. The reaction mixture was poured into ice water (700 ml) and extracted with petroleum ether. The organic layer was successively washed with water, aqueous potassium carbonate solution and then water, dried over anhydrous sodium sulfate and concentrated. The resulting residue was distilled to give the title compound (9.7 g), bp 95-108-C/30mmHg. NMR (d in CDC13), 7.94 (ddd, Jn6.7, 7.6, 9.0 Hz) Reference example 7. 3-Chloro-2-cyclopropylamino-4 , 5-difluoro- nitrobenzene A solution of cyclopropylamine (2.8 g) and triethylamine (5.1 g) in anhydrous toluene (20 ml) was added dropwise to a solution of 3-chloro-2,4,5-trifluoronitrobenzene (9.7 g) in anhydrous toluene (30 ml) at 3-50C during 40 minutes with stirring. After stirring for 3 hours at the same temperature, the reaction mixture was poured into ice water (150 ml) and extracted with dichloromethane. The organic layer was washed with water, dried over anhydrous sodium sulfate and then concentrated. The resulting residue was purified by silica gel chromatography using n-hexane-dichloromethane as an eluent to give the title compound (4.4 g) as red oil. NMR (d in CDC13), 0.5-1.0 (4H, m, EMI15.1 3.0-3.2 (1H, m, EMI15.2 7.19 (1H, s, NH), 7.85 (1H, dd, J=8.2, 9.9 Hz, 5-H). Reference example 8. N-( 2-Chloro-3, 4-difluoro-6-nitrophenyl)-N- cyclopropylacetamide To 3-chloro-2-cyclopropylamino-4 ,5-difluoronitrobenzene (4.4 g) was added acetic anhydride (15 ml) with one portion and the mixture was stirred for 30 minutes at room temperature, and then poured into ice water (100 ml). Excessive acetic anhydride was decomposed by potassium carbonate powder, and then the mixture was allowed to stand overnight at 5-C. The resulting precipitate was collected by filtration and recrystallized from ethyl acetate-n-hexane toigive the title compound (2.7 g), mp 98-99.5-C. Analysis (%) for C11H9ClF2N2O35 Calcd. (Found): C, 45.46 (45.56); H, 3.12 (3.00); N, 9.64 (9.69). Reference example 9. N-(2-Chloro-3,4-difluorophenyl)-N-cyclo propylacetamide A mixture of N-( 2-chloro-3 , 4-difluoro-6-nitrophenyl)-N- cyclopropylacetamide (2.7 g) and 10% palladium-charcoal (0.5 g) in ethanol (50 ml) was hydrogenated for 40 minutes at 2-3'C under atmospheric pressure in an ice bath. The catalyst was filtered off and the filtrate was concentrated. Then, the resulting crystalline residue was dried in vacuo at room temperature for 10 hours. A solution of the residue in anhydrous dimethylformamide (15 ml) was added dropwise to a solution of tert-butyl nitrite (1.72 g) in anhydrous dimethylformamide (10 ml) at 50-52*C during 13 minutes. The reaction mixture was stirred at the same temperature for 5 minutes, then poured into ice water and extracted with ether. The organic layer was successively washed with water, dilute hydrochloric acid and water, dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by silica'gel chromatography using n-hexane-ethyl acetate as an eluent and recrystallized from petroleum ether to give the title campound (0.44 g), mp 60.5-61.5-C. Analysis (*) for CllHloClF2NO, Calcd. (Found): C, 53.78 (53.87); H, 4.10 (4.02); N, 5.70 (5.78). Reference example 10. N-Cyclopropyl-2-chloro-3. 4-difluoroaniline A solution of N-(2-chloro-3,4-difluorophenyl9-Ncyclopropyl- acetamide (0.44 g) in 20% diluted hydrochloridfecid (7 ml) was heated at 80-100 C for 6 hours with stirring, and then cooled. The reaction mixture was poured into ice water, alkalized with aqueous sodium hydroxide and extracted with ether. The ether layer was washed with water, dried over anhydrous sodium sulfate and then concentrated. The resulting residue was purified by silica gel chromatography using petroleum ether as an eluent to give the title compound (100 mg) as orange oil. Reference example 11. Ethyl 8-chloro-1-cyclopropyl-6,7-difluoro 1,4-dihydro-4-oxo-3-quinolinecarboxylate A mixture of N-cyclopropyl-2-chloro-3,4-difluoroaniline (100 mg) and diethyl ethoxymethylenemalonate (100 mg) was stirred for 10.5 hours at 100-135-C with removal of generated ethanol by flowing nitrogen gas, and then cooled. Polyphosphoric acid (1 g) was mixed with this, and the mixture was stirred for 3 hours at 125-135-C. After cooling, the reaction mixture poured into ice water, extracted with chloroform. The organic layer was successively washed with aqueous potassium carbonate and water, dried over anhydrous sodium sulfate and then concentrated. The resulting residue was purified with silica gel thin layer chromatography using ether as an eluent to give the title compound (11 mg) as colorless needles, mp 160-162.5-C. NMR (d in CDC13), 1.0-1.5 (4H, m, EMI17.1 1.40 (3H, t, J=7.0 Hz, ¯CH3), 4.1-4.4 (1H, m, EMI17.2 4.38 (2H, q, J=7.O Hz, -CH2 CH3), 8.22 (1H, dd, Jx8.8, 9.7 Hz, 5-H), 8.66 (1H, s, 2-H) Reference example 12. 2,3,4-Trichloro-5-fluoroaniline To a suspension of iron powder (54.6 g) in water (60 ml), with vigorous stirring at 50-6OC, was slowlydedded concentrated hydrochloric acid (6.7 ml). After ethanol (150 ml) was mixed, 2,3,4-trichloro-5-fluoronitrobenzene (75.1 g) was added portionwise to the suspension at 60-70-C during an hour. After stirring for an hour at 80 C. the hot reaction mixture was filtered and the insoluble material was successively washed with hot ethanol (100 ml) and benzene (300 ml). The filtrate and washings were combined and mixed with ice water. The resulting organic layer was collected and the water layer was extracted with benzene (200 ml). The organic layers were washed with water, dried over anhydrous sodium sulfate and then concentrated. The resulting residue was recrystallized from n-hexane to give the title compound (58.6 g) as light brown needles, mp 118-120*C. Reference example 13. 2,3,4-Trichloro-5-fluorobenzonitrile To a suspension of 2,3,4-trichloro-5-fluoroaniline (43.8 g) in concentrated hydrochloric acid (300 ml) with vigorous stirring was added sodium nitrite (21.1 g) in water (50 ml) at -2"'0C for 20 minutes. After stirred for 30 minutes, the mixture was poured into ice water (300 ml) containing sodium tetrafluoroborate (67.2 g), stirred vigorously and then allowed to stand for 30 minutes in an ice bath. The resulting precipitate was collected by filtration and successively washed with chilled water and ether. This faint yellow precipitate was added portionwise during 30 minutes to a solution of cuprous cyanide (36.5 g), potassium cyanide (53.0 g) and sodium carbonate (11.1 g) in water (300 ml) with vigorous stirring at room temperature. After the mixture was stirred for 30 minutes, benzene (300 ml) was added to the suspension and then the mixture was stirred forl15 minutes. The insoluble material was collected by filtration, and washed with benzene (150 ml). The filtrate and washings were combined and successively washed with 20% aqueous potassium cyanide and water, dried over anhydrous sodium sulfate and then concentrated. The resulting residue was recrystallized from n-hexane to give the title compound (27 g) as light brown needles, mp 97-99-C. Reference example 14. 3-Chloro-2 ,4 ,5-trifluorobenzonitrile To a solution of potassium fluoride (31.7 g) in dimethyl sulfoxide (100 ml) with stirring at 130C was added 2,3,4-tri chloro-5-fluorobenzonitrile (15 g) and then the mixture was stirred for 1.5 hours at 1400C. After cooling, the reaction mixture was poured into ice water (300 ml) and extracted with dichloromethane. The organic layer was washed with water, dried over anhydrous sodium sulfate and concentrated to give the title compound (11.9 g) as pale brown oil. Reference example 15. 3-Chloro-2,4,5-trifluorobenzamide A solution of 3-chloro-2,4,5-trifluorobenzonitrile (11.9 g) in 30% hydrogen bromide-acetic acid (150 ml) was refluxed for 80 minutes, poured into ice water (350 ml) and extracted with ether. The ether layer was successively washed with 1N-potassium hydroxide solution and water, dried over anhydrous sodium sulfate and concentrated. The residue was purified by silica gel chromatography eluting with n-hexane-ethyl acetate to give the title compound (3.97 g), mp 110-113.5-C. Reference example 16. 3-Chloro-2,4,5-trifluorobenzoic acid A mixture of 3-chloro-2,4,5-trifluorobenzamide (3.97 g) and 18N-sulfuric acid (20 ml) was stirred at 125-5'C for 9 hours, and then poured into ice water (10C ml). After standing overnight, the resulting precipitate was collected by filtration. The mother liquor was extracted with ether, and the ether layer was dried over anhydrous sodium sulfate, then concentrated and mixed to the precipitate. A solution of the above precipitate and residue in dichloromethane (150 ml) was filtered through a celite pad and the filtrate was concentrated to give the title compound (2.38 g), mp 115-116.5iC. Analysis (%) for C7H2ClF302, Calcd. (Found): C, 39.93 (40.18); H, 0.96 (0.80). Reference example 17. 3-Chloro-25455-trifluorobenzoyl chloride A solution of the 3-chloro-2,4,5-trifluorobenzoic acid (2.38 g) in thionyl chloride (10 ml) was reluxed for 2.5 hours, and then concentrated. The resulting residue was purified by distillation in nitrogen atomosphere to give the title compound (1.99 g), bp 88 C/19 mmHg. Reference example 18. Diethyl 3-chloro-2,4,5-trifluorobenzoyl malonate Magnesium turnings (0.22 g) and carbon tetrachloride (0.1 ml) was added to absolute ethanol (1.5 ml). To the stirring suspension was added dropwise a solution of diethyl malonate (1.4 g) and absolute ethanol (2 ml) in toluene (6 ml) during 28 minutes at 47-60*C. The mixture was stirred for 80 minutes, and then cooled in an acetone-dry ice bath. A solution of 3-chloro2,4,5-trifluorobenzoyl chloride (1.99 g) in anhydrous toluene (2 ml) was added dropwise to the resulting solution at -12"'-80C during 13 minutes. The mixture was stirred for 2 hours at -10#- 5µC allowed to stand overnight at room temperature, and then mixed with ice water (6 ml) containing concentrated sulfuric acid (0.4 ml). The resulting organic layer was collected and the water layer was extracted with toluene. The combined organic layer was washed with water, dried over anhydrous sodium sulfate and then concentrated to give the title compound (3.05 g) as pale yellow oil. Reference example 19. Ethyl 3-chloro-2, 4, 5-trifluorobenzoyl- acetate To an emulsion of diethyl 3-chloro-2,4,5-trifluorobenzoyl: malonate (3.05 g) in water (4 ml) was added p-toluenesulfonic acid (4 mg) and refluxed for 4 hours with vigorous stirring. After cooling, the reaction mixture was extracted with dichloromethane. The organic layer was washed with water, dried over anhydrous sodium sulfate and concentrated. The residue was recrystallized from ether-n-hexane to give the title compound (1.22 g), mp 80-83eC. Analysis (%) for CllH8ClF303, Calcd. (Found): C, 47.08 (46.96); H, 2.87 (2.77). Reference example 20. Ethyl 2-(3-chloro-2,4,5-trifluorobenzoyl) 3 -ethoxyacrylate A mixture of ethyl 3-chloro-2,4,5-trifluorobenzoylacetate (1.22 g), ethyl orthoformate (0.97 g) and acetic anhydride (1.12 g) was stirred at 118-143"C for 3 hours and then concentrated to give the title compound (1.4 g) as yellow oil. Reference example 21. Ethyl 2-(3-chloro-2,4,5-trifluorobenzoyl) 3 -cyclopropylaminoacrylate To a solution of ethyl 2-(3-chloro-2,4,5-trifluorobenzoyl)- 3-ethoxyacrylate (1.4 g) in absolute ethanol (3 ml) was added a solution of cyclopropylamine (0.26 g) in absolute ethanol (2 ml) at 5-10'C during 15 minutes. The mixture was allowed to stand at 50C for 1.5 hours and then stirred at room temperature for an hour. The resulting precipitate was collected by filtration. The filtrate was concentrated, mixed with the precipitate and then recrystallized from petroleum ether to give the title compound (1.09 g), mp 84-85.50C. Analysis (%) for C15Hl3ClF3NO3, Calcd. (Found): C, 51.81 - (51.76); H, 3.77 (3.74); N, 4.03 (4.03). Reference example 22. Ethyl 8-chloro-1-cyclopropyl-6,7-difluoro- 1,4-dihydro-4-oxo-3-quinolinecabylate To a solution of ethyl 2-(3-chloro-2,4,5-trifluorobenzoyl)3-cyclopropylaminoacrylate (1.09 g) in anhydrous dimethylformamide (5 ml) was added sodium fluoride (0.21 g). The mixture was stirred at 130-156'C for 3.5 hours, and then poured into ice water (50 ml) and the resulting precipitate was collcted by filtration, washed with water and recrystallized from ethyl acetate to give the title compound (0.96 g), mp 158-159-C. Analysis (%) for C C15Hl2ClF2N03, Calcd. (Found): C, 54.98 (54.96); H, 3.69 (3.57); N, 4.27 (4.25). Reference example 23. 8-Chloro-1-cyclopropyl-6,7-difluoro-1,4- dihydro-4-oxo-3-quinolinecarboxylic acid A mixture of ethyl 8-chloro-1-cyclopropyl-6,7-difluoro-1,4dihydro-4-oxo-3-quinolinecarboxylate (0.24 g), acetic acid (2 ml), water (1.5 ml) and concentrated sulfuricecid (0.25 ml) was refluxed for an hour, and then poured into ice water. The resulting precipitate was collected by filtration and successively washed with water and ether to give the title compound (0.17 g), mp 194-195iC. Analysis (t) for C13H8ClF2N03, Calcd. (Found): C, 52.11 (52.00); H, 2.69 (2.53) N, 4.67 (4.64). Example 13. 8-Chloro-l-cyclorpropyl-6-fluoro-1,4-dihydro-7-(3 ethylaminomethyl-l-pyrrolidinyl)-4-oxo-3-quinoline carboxylic acid A mixture of 8-chloro-1-cyclopropyl-,67-difluoro-1,4- dihydro-4-oxo-3-quinolinecarboxylic acid (0.4 g), 3-ethylaminomethylpyrrolidine (0.19 g) and DBU (0.2 g) in acetonitrile (4 ml) was heated to reflux for 2 hours and then allowed to stand overnight during which time a crystal separated. The crystal, title compound, was filtered and recrystallized from chloroformmethanol giving colorless prisms (0.39 g), mp 250-252iC (decompd.). Analysis (%) for C20H23ClFN303*1/2 H2O, Calcd. (Found): C, 57.62 (57.48); H, 5.80 (5.52); N, 10.08 (10.07). Example 14. 8-Chloro-l-cyclopropyl-6-fluoro-1,4-dihydro-7- (3-aminomethyl-1-pyrrolidinyl)-4-oxo-3-quinoline carboxylic acid hydrochloride A mixture of 8-chloro-1-cyclopropyl-6,7-difluoro-1,4- dihydro-4-oxo-3-quinolinecarboxylic acid (0.35 g), 3-aminomethylpyrrolidine (0.13 g), and DBU (0.19 g) in acetonitrile (4 ml) was refluxed for 2 hours and then allowed to stand overnight. A solid separated was collected by filtration and recrystallized from chloroform-methanol-n-hexane giving the crystalline product which was further purified by silica gel column chromatography using chloroform-methanol-concentrated aqueous ammonia (20 : 6 1) as eluent to obtain the free form (188 mg) of the title compound. The compound was dissolved in a mixture of methanol and concentrated to dryness. The residue was successively recrystallized from ethanol-acetonitrile and ethanol to give the title compound (14 mg) as pale yellow prisms, mp 238-247"C (decompd.). Analysis (%) for C18H19C1FN303.HCl, Calcd. (Found): C, 51.94 (51.63); H, 4.84 (5.01); N, 10.09 (9.85). Example 15. 7-(3-Aminomethyl-l-pyrrolidinyl)-8-bromo-1-cyclo propyl-6-fluoro-1,4-dihydro-4-oxo-3-quinoline- carboxylic acid A mixture of 8-bromo-1-cyclopropyl-6, 7-difluoro-1, 4- dihydro-4-oxo-3-quinolinecarboxylic acid (200 mg), 3-aminomethylpyrrolidine (60 mg) and DBU (90 mg) in acetonitrile (3 ml) was stirred for an hour under refluxing and then for further 4 hours at room temperature. The crystals which separated were collected and recrystallized from dichloromethane to give the title compound (40 mg) as pale yellow prisms, mp 222-230-C (decompd.). Analysis (%) for C18HlgBrFN303.H2O, Calcd. (Found): C, 48.80 (48.80); H, 4.79 (4.74); N, 9.50 (9.45). In this example, the starting material is also novel and it is synthesized by following process. Reference example 24. 2-Bromo-3-chloro-4-fluoro-6-nitroaniline Into a solution of 3-chloro-4-fluoro-6-nitroaniline (200,3 g) in acetic acid (1.5 litter) was added bromine (339 g) during a period of 80 minutes at 50'C under stirring and stirred for further 2 hours. The reaction mixture was poured into ice water (3 litter) and the resulting precipitate was collected by filtration, washed with water and added to a mixture of concentrated hydrochloric acid (300 ml) and ethanol (1.2 litter). The mixture was refluxed for 8.5 hrs. After cooling, the precipitate was collected by filtration and washed with water and dried. The title compound thus obtained weighed 235.6 g as yellow needles, mp 146-147iC. Reference example 25. 3-Bromo-2,4-dichloro-5-fluoronitrobenzene To a mixture of anhydrous cupric chloride (147 g) and 2 bromo-3-chloro-4-fluoro-6-nitroanilins (235.6 g) in anhydrous acetonitrile (1.5 litter) was added tert-butylnitrite (135.2 g) at 600C during 70 minutes. The reaction mixture was poured into ice-chilled diluted hydrochloric acid (1.5 litter) and extracted with benzene. The organic layer was successively washed with ice-chilled diluted hydrochloric acid and water saturated with sodium chloride, dried over anhydrous sodium sulfate and concentrated. The resulting residue was purified by distillation to give the title compound(218.8 g), bp 78-1170C/2 mmHg. The oil was crystallized from methanol to give yellow prisms, mp 65.5 67.5C. Reference example 26. 3-Bromo-2,4-dichloro-5-fluoroaniline To a suspension of iron powder (135.4 g) in water (140 ml), with vigorous stirring at 50-600C, was slowly added concentrated hydrochloric acid (18 ml). After ethanol (350 ml) was mixed, 3bromo-2,4-dichloro-5-fluoronitrobenzene (218.8 g) was added portionwise to the suspension at 52-76-C during an hour. After stirring for 75 minutes at the same temperature, the hot reaction mixture was filtered after adding benzene (500 ml) and the insoluble material was successively washed with hot ethanol (100 ml) and benzene (200 ml). The filtrate and washings were combined. The organic layers were washed with water saturated with sodium chloride, dried over anhydrous sodium sulfate and then concentrated. The resulting residue was recrystallized from ethanol-water to give the title compound (141.6 g) as light brown needles, mp 126-129.5-C. Reference example 27. 3-Bromo-2,4-dichloro-5-fluorobenzonitrile To a suspension of 3-bromo-2,4-dichloro-5-fluoroaniline (141.6 g) in concentrated hydrochloric acid (900 ml) with vigorous stirring was added sodium nitrite (56.6 g) in water (120 ml) at -2 s0iC for 40 minutes. After stirred for 30 minutes, the mixture was poured into ice water (700 ml) containing sodium * tetrafluoroborate (180 g), stirred vigorously for 20 minutes and then allowed to stand for 15 minutes in an ice bath. The resulting precipitate was collected by filtration and washed with chilled water. The wet crude tetrafluoroborate thus obtained weighed 270.8 g. The borate was added portionwise during 45 minutes to a solution of cuprous cyanide (98 g), potassium cyanide (142.4 g) and sodium carbonate (29 g) in water (800 ml) with vigorous stirring at 9-10'C. After the mixture was stirred for 2 hours at room temperature, benzene (700 ml) and potassium cyanide (71 g) were added to the suspension and then the mixture was stirred for 30 minutes. The insoluble material was collected by filtration, and washed with benzene (300 ml x 2). The filtrate and washings were combined and washed five times with water saturated with sodium chloride, -dried over anhydrous sodium sulfate and then concentrated. The resulting residue was recrystallized from ethanol to give the title compound (75.5 g) as red brown prisms, mp 110.5-112.5eC. Reference example 28. 3-Bromo-2, 4 ,5-trifluorobenzonitrile To a solution of potassium fluoride (123 g) in dimethyl sulfoxide (400 ml) with stirring at 1330C was added 3-bromo-2,4 dichloro-5-fluorobenzonitrile (68.4 g) and then the mixture was stirred for 5 hours and 20 minutes at 1300C. After cooling, the reaction mixture was poured into ice water (1 litter) and extracted with benzene. The organic layer was washed with water saturated with sodium chloride, dried over anhydrous sodium sulfate and distilled to give the title compound (15.7 g) as colorless oil, bp 82.5-C/13 mmHg - 80.O'C/lmmHg. Reference example 29. 3-Bromo-2,4,5-trifluorobenzoic acid A mixture of 3-bromo-2,4,5-trifluorobenzonitrile (13.9 g) in concentrated sulfuric acid (8 ml) was heated for 20 minutes on an oil bath (100-C), poured into ice water (350 ml). The resulting precipitate was collected by filtration and washed with water. The filtrate and washings were extracted 3 times with dichloromethane. The dichloromethane layer was washed with water saturated with sodium chloride, dried over anhydrous sodium sulfate and concentrated to give the residue. The combined mixture of the precipitate obtained previously and the residue was purified by silica gel chromatography eluting with dichloromethane + dichloromethane z methanol (10 s 1) to give 3-bromo2,4,5-trifluorobenzamide (8.7 g). A mixture of 3-bromo-2,4,5-trifluorobenzamide (8.7 g) and 18N-sulfuric acid (50 ml) was stirred at 100'C for 4 hours, and then poured into ice water (200 ml). The resulting precipitate was collected by filtration and recrystallized from dichloromethane-n-hexane to give the title compound (6.9 g), mp 125 127-C. Reference example 30. 3-Bromo-2,4,5-trifluorobenzoyl chloride A solution of the 3-bromo-2,4,5-trifluorobenzoic acid (2.5 g) in thionyl chloride (10 ml) was refluxed for 2.5 hours, and then concentrated. The resulting residue was purified by distil lation through Widmer fractionating column to give the title compound (2.3 g), bp 98-102iC/18 mmHg. Reference example 31. Diethyl 3-bromo-2,4,5-trifluorobenzoyl malonate Magnesium turnings (0.22 g) and carbon tetrachloride (0.1 ml) was added to absolute ethanol (1.5 ml). To the stirring suspension was added dropwise a solution of diethyl malonate (1.4 g) and absolute ethanol (2 ml) in toluene (6 ml) during 25 minutes at 50-600C. The mixture was stirred for 40 minutes, and then cooled. A solution of 3-bromo-2,4,5-trifluorobenzoyl chloride (2.27 g) in anhydrous toluene (3 ml) was added dropwise to the solution at -8"'-4.50C during 28 minutes. The mixture was stirred for 2 hours and then mixed with ice-chilled diluted sulfuric acid. The resulting organic layer wascollected and the water layer was extracted with toluene (6 ml x 4). The combined organic layer was washed with water, dried over anhydrous sodium sulfate and then concentrated to give the title compound (3.25 g) as pale yellow oil. Reference example 32. Ethyl 3-bromo-2,4,5-trifluorobenzoyl acetate To an emulsion of diethyl 3-bromo-2,4,5-trifluorobenzoylmalonate (3.25 g) in water (4 ml) was added p-toluenesulfonic acid (4 mg) and refluxed for 3 hours with vigorous stirring. After cooling, the reaction mixture was extracted with dichloro methane (8 ml x 4). The organic layer was washed with water saturated with sodium chloride, dried over anhydrous sodium sulfate and concentrated. The residue was recrystallized from dichloromethane-n-hexane to give the title compound (1.51 g), mp 85-88-C. Reference example 33. Ethyl 2-(3-bromo-2,4,5-trifluorobenzoyl) 3-ethoxyacrylate A mixture of ethyl 3-bromo-2,4,5-trifluorobenzoylacetate (1.5 g), ethyl orthoformate (1.0 g) and acetic anhydride (1.2 g) was stirred at 1300C for 4.5 hours and then concentrated to give the title compound (1.75 g) as yellow oil. Reference example 34. Ethyl 2-(3-bromo-2,4,5-trifluorobenzoyl) 3-cyclopropylaminoacrylate To a solution of ethyl 2-(3-bromo-2,4,5-trifluorobenzoyl)-3ethoxyacrylate (1.75 g) in absolute ethanol (5 ml) was added a solution of cyclopropylamine (0.32 g) in absolute ethanol (2 ml) under ice-cooling during 30 minutes. The mixture was stirred at 5-200C for 2.5 hours and concentrated. The residue was recrystallized from petroleum ether to give the title compound (1.36 g), mp 74-76'C. Reference example 35. Ethyl 8-bromo-1-cycl6propyl-6,7-difluoro- 1,4-dihydro-4-oxo-3-quinolinecarboxylate To a solution of ethyl 2-(3-bromo-2,4,5-trifluorobenzoyl)-3cyclopropylaminoacrylate (1.35 g) in anhydrous dimethylformamide (5 ml) was added sodium fluoride (0.23 g). The mixture was stirred at 97-108iC for 7.5 hours, and then poured into ice water (50 ml) and the resulting precipitate was collected by fil tration, washed with water and recrystallized from dichloromethane-n-hexane to give the title compound (1.05 g), mp 163.5 1680C as colorless prisms. Reference example 36. 8-Bromo-l-cyclopropyl-6,7-difluoro-1,4- dihydro-4-oxo-3-quinolinecarboxylic acid A mixture of ethyl 8-bromo-1-cyclopropyl-6,7-difluoro-1,4- dihydro-4-oxo-3-quinolinecarboxylate (1.0 g), acetic acid (4 ml), water (3 ml) and concentrated sulfuric acid (0.5 ml) was heated on an oil bath (90-100'C) for an hour under stirring, then for an hour at room temperature and poured into ice water (20 ml). The resulting precipitate was collected by filtration and washed with water to give the title compound (0.82 g), mp 224225.50C. Example 16. 8-Bromo-l-cyclopropyl-6-fluoro-1,4-dihydro-7-(3 methylaminome thyl-l-pyrrolidinyl ) -4-oxo-3-quinoline- carboxylic acid A mixture of 8-bromo-1-cyclopropyl-6,7-dtiuoro-1,4-dihydro- 4-oxo-3-quinolinecarboxylic acid (200 mg), 3-methylaminomethylpyrrolidine (FO mg) and DBU (100 mg) in acetonitrile (3 ml) was stirred for an hour under refluxing and then for further 3 hours at room temperature. The crystals which separated were collected and recrystallized from chloroform-methanol-ammonia to give the title compound (160 mg) as pale yellow prisms, mp 242.5-2460C (decompd.). Analysis (z) for C19H21BrFN3O3, Calcd. (Found): C, 52.07 (52.28); H, 4.83 (4.85); N, 9.59 (9.61). Example 17. 8-Bromo-l-cyclopropyl-7-(3-ethylaminomethyl-1 pyrrodinyl)-6-fluoro-1,4-dihydro-1,4-dihydro-4-oxo-3 quinolinecarboxylic acid A mixture of 8-bromo-1-cyclopropyl-6,7-difluoro-1,4-dihydro- 4-oxo-3-quinolinecarboxylic acid (280 mg), 3-ethylaminomethylpyrrolidine (90 mg) and DBU (90 mg) in acetonitrile (3 ml) was stirred for an hour under refluxing and then for further 3 hours at room temperature. The crystals which separated were collected and recrystallized from chloroform-methanol-ammonia to give the title compound (150 mg) as colorless prisms, mp 258-260'C (decompd.) Analysis (8) for C20H23BrFN303, Calcd. (Found): C, 53.11 (53.54); H, 5.12 (5.11); N, 9.29 (9.43). Example 18. Ethyl 8-chloro-1-cyclopropyl-7-(3-ethylaminoethyl- l-pyrrolidinyl)-6-fluoro-1,4-dihydro-4-oxo-3- quinolinecarboxylate A mixture of ethyl 8-chloeo-1-cyclopropyl-6,7-difluoro-1,4- dihydro-4-oxo-3-quinolinecarboxylate (500 mg),--acetonitrile (5 ml), 3-ethylaminomethyipyrrolidine (296 mg) and DBU (233 mg) was refluxed for 5 hours and then concentrated to give the residue, to which ice water (20 ml) was added. The mixture was extracted with chloroform. The extract was washed with water, dried over anhydrous sodium sulfate and concentrated to give the title compound (800 mg) as brown oil. IR (cm ): 3010, 1720, 1610, 1450, 1315. NMR (d in CDC13)s 0.9-1.3 (4H, m, EMI31.1 1.12 (3H, t, -N CH2CH3), 1.39 (3H, t, -O-CH2CH3), 1.5-1.9 (1H, m, EMI31.2 2.0-2.2 (1H, m, EMI31.3 2.2-2.55 (1H, m, EMI31.4 2.68 (4H, m, EMI31.5 3.3-3.7 (4H, m, EMI32.1 4.17 (1H, m, EMI32.2 4.37 (2H, q, CH2CH3), 7.93 (1H, d, J=13.6 Hz, 5-H), 8.60 (1H, s, 2-H). Example 19. 8-Chloro-l-cyclopropyl t 3-ethylaminomethyl-1 pyrrolidinyl)-6-fluoro-1,4-dihydro-4-oxo-3 quinolinecarboxylic acid A mixture of ethyl 8-chloro-1-cyclopropyl-7-(3-ethylamino- methyl-1-pyrrolidiyl)-6-fluoro-1,4-dihydro-4-oxo-3-quinolinecarboxylate (800 mg) and 1N sodium hydroxide (8 ml) was refluxed for 1 1/3 hours and concentrated. The residue was purified by silica gel column chromatography (CHCl3 t MeOH - 4:1) to give yellow crystalline powder. The powder was suspended in acetonitrile, then collected by filtration and recrystallized from chloroform-methanol to give the title compound (130 mg) as yellow prisms, mp 248-251'C (decompd.). Analysis (%) for C20H23ClFN3O3-1/4 H2O, Calcd.(Found): C, 58.25 (58.46); H, 5.74 (5.65); N, 10.19 (10.26). Example 20. 8 -Chloro-1 -cyclopropyl- 6 -fluoro-A -dihydro-7 - (3 - dimethyldaminomethylaminomethyl-1-pyrrolidinyl)-4-oxo-3 quinolinecarboxylic acid A mixture of 8-chloro-1-cyclopropyl-6,7-difluoro-1,4- dihydro-4-oxo-3-quinolinecarboxylic acid (500 mg), 3-dimethylaminomethylpyrrolidine (330 mg), DBU (250 mg) and acetonitrile (5 ml) was refluxed for an hour under stirring, then allowed to stand for overnight and concentrated. To the resulting residue was added methanol and the mixture was filtered to collect the crystalline product which was recrystallized from chloroformmethanol to give the title compound (430 mg) as yellow prisms, mp 175-176-C. Analysis (%) for C20H23C1FN303, Calcd. (Found): C, 58.90 (59.02), H, 5.68 (5.70) N, 10.30 Example 21. 1-Cyclopropyl-6,8-difluoro-1,4-dihydro-7-(3- dimethylaminomethyl-l-pyrrolidinyl)-4-oxo-3- quinolinecarboxylic acid A mixture of 1-cyclopropyl-6,7,8-trifluoro-1,4-dihydro-4- oxo-3-quinolinecarboxylic acid (500 mg), 3-dimethylaminomethylpyrrolidine (340 mg), DBU (270 mg) and acetonitrile (5 ml) was refluxed for an hour under stirring, allowed to stand for overnight and filtered to collect the crystalline product which was washed with acetonitrile and ether successively to give the title compound (420 mg) as pale yellow needles, mp 178-181'C. Analysis (8) for C20H23F2N3035 Calcd. (Found): C, 61.37 (61.03); H, 5.92 (5.95); N, 10.74 (10.76). Experiment-l In vitro antibacterial activity The in vitro antibacterial activity against gram-positive and gram-negative bacteria has been investivated. The minimum inhibitory concentration (MIC) was determined in accordance with the method recommended by Japan Society of Chemotherapy. The result of the test was shown in Table 1 and 2. In these experiments, the 8-chloro or 8-bromo compounds described in Example 12 to 20 have antibacterial properties which are even more marked than those of their 8-hydrogen or 8-fluoro homologues, as shown in Table 1 and 2, against aerobic grampositive bacteria and anaerobic bacteria. Table 1-1 In vitro antibacterial activity standart strain) EMI34.1 <tb> <SEP> HIC( g/ml) <tb> <SEP> Organism <SEP> (106 <SEP> cells/ml) <SEP> Gram <tb> <SEP> Exp1 <SEP> Exp2 <SEP> Exp4 <SEP> Exp6 <SEP> Exp7 <tb> Bacillussubtills <SEP> PC:219 <SEP> 0,025 <SEP> 0,0125 <SEP> 0,025 <SEP> O,05 <SEP> 0,025 <tb> Staphilococcus <SEP> aureus <SEP> 209 <SEP> P <SEP> + <SEP> 0.05 <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <tb> S. <SEP> aureus <SEP> Smith <SEP> ¯¯¯¯¯¯¯¯¯¯¯¯¯ <SEP> <SEP> + <SEP> 0.05 <SEP> 0.025 <SEP> 0.05 <SEP> 0.20 <SEP> 0.10 <tb> S. <SEP> aureus <SEP> IID <SEP> 670 <SEP> (Terajima) <SEP> t <SEP> =.05 <SEP> 0.025 <SEP> 0.025 <SEP> 0.05 <SEP> 0.10 <tb> S. <SEP> epidermidin <SEP> IID <SEP> 866 <SEP> + <SEP> 0.05 <SEP> 0.05 <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <tb> Streptococcus <SEP> pyogenes <SEP> S-8 <SEP> ¯ <SEP> t <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.10 <SEP> 0.39 <tb> S. <SEP> pyogenes <SEP> IID <SEP> 692 <SEP> t <SEP> t <SEP> 0.05 <SEP> 0.20 <SEP> 0.05 <SEP> 0.20 <SEP> 0.39 <tb> S. <SEP> pneumoniae <SEP> llD <SEP> 552 <SEP> + <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.10 <SEP> 0.39 <tb> S. <SEP> faecalis <SEP> IID <SEP> 682 <SEP> t <SEP> 0.05 <SEP> 0.20 <SEP> 0.05 <SEP> 0.39 <SEP> 0.39 <tb> Eschericbia <SEP> coli <SEP> NIHJ <SEP> JC-2 <SEP> - <SEP> 0.025 <SEP> 0.025 <SEP> 0.0125 <SEP> 0.05 <SEP> 0.05 <tb> E. <SEP> coli <SEP> ATCC <SEP> 10536 <SEP> - <SEP> 0.02 <SEP> 0.025 <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <tb> E. <SEP> coli <SEP> Ml. <SEP> 4707 <SEP> - <SEP> 0.02- <SEP> 0.025 <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <tb> Proteus <SEP> vulqaris <SEP> IFO <SEP> 3167 <SEP> - <SEP> 0.025 <SEP> 0.025 <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <tb> P. <SEP> mirabilis <SEP> llD <SEP> 994 <SEP> - <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <tb> Morqanella <SEP> morganii <SEP> IID <SEP> 602 <SEP> - <SEP> 0.10 <SEP> 0.10 <SEP> 0.10 <SEP> 0.39 <SEP> 0.39 <tb> Enterobacter <SEP> cloacae <SEP> IID <SEP> 977 <SEP> - <SEP> 0.10 <SEP> 0.10 <SEP> 0.05 <SEP> 0.20 <SEP> 0.39 <tb> Citrobacter <SEP> freundii <SEP> IID <SEP> 976 <SEP> - <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.10 <tb> Klebsiella <SEP> pneumoniae <SEP> KY(GN)6645 <SEP> - <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.10 <tb> K. <SEP> pneumoniae <SEP> 1-220S <SEP> - <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.20 <SEP> 0.20 <tb> Salmonella <SEP> entritidis <SEP> IID <SEP> 604 <SEP> - <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> <SEP> 0.20 <SEP> 0.20 <tb> Shigella <SEP> sonnei <SEP> llD <SEP> 969 <SEP> - <SEP> 0.025 <SEP> 0.025 <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <tb> Yersinia <SEP> enterocolitica <SEP> IID <SEP> 981 <SEP> - <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.20 <SEP> 0.39 <tb> Serratia <SEP> marcescens <SEP> IID <SEP> 618 <SEP> - <SEP> 0.20 <SEP> 0.10 <SEP> 0.10 <SEP> 0.39 <SEP> 0.39 <tb> S. <SEP> marcescens <SEP> GN <SEP> 7577 <SEP> - <SEP> 0.7F <SEP> 1.56 <SEP> 0.78 <SEP> 3.13 <SEP> 6.25 <tb> Pseudomonas <SEP> aeruginosa <SEP> V-1 <SEP> - <SEP> 0.35 <SEP> 0.78 <SEP> - <SEP> 0.78 <SEP> 1.56 <tb> P. <SEP> aeruginosa <SEP> IFO <SEP> 12689 <SEP> - <SEP> 0.35 <SEP> 1.56 <SEP> 0.39 <SEP> 0.78 <SEP> 3.13 <tb> P. <SEP> aeruginosa <SEP> IID <SEP> 1210 <SEP> - <SEP> 0.78 <SEP> 3.13 <SEP> 0.78 <SEP> 3.13 <SEP> 3.13 <tb> Acinetobacter <SEP> anitratus <SEP> IID <SEP> 876 <SEP> - <SEP> 0.10 <SEP> 0.05 <SEP> 0.05 <SEP> 0.78 <SEP> 0.78 <tb> Alcaligenes <SEP> faecalis <SEP> 0104002 <SEP> - <SEP> 0.39 <SEP> 0.10 <SEP> 0.78 <SEP> 1.56 <SEP> - <SEP> 1.56 <tb> bacteriodes <SEP> fragilis <SEP> GM <SEP> 7000 <SEP> - <SEP> 1,56 <SEP> - <SEP> 6.25 <SEP> 25 <tb> B. <SEP> fragilis <SEP> 0558 <SEP> - <SEP> 0.39 <SEP> 0.78 <SEP> 0.78 <SEP> 6.25 <SEP> 25 <tb> 8. <SEP> fragilis <SEP> 25285 <SEP> - <SEP> 0.39 <SEP> 0.78 <SEP> 0.78 <SEP> 12.5 <SEP> 25 <tb> Fusobacterium <SEP> varium <SEP> KYA <SEP> 8501 <SEP> - <SEP> 0.78 <SEP> 1.56 <SEP> 1.56 <SEP> 25 <SEP> 12.5 <tb> Clostridium <SEP> perfringens <SEP> KYA13123 <SEP> + <SEP> 0.39 <SEP> 0.39 <SEP> - <SEP> 3.13 <SEP> 0.20 <tb> C. <SEP> ramosum <SEP> + <SEP> 0.78 <SEP> 0.78 <SEP> 0.78 <SEP> 12.5 <SEP> 6.25 <tb> C. <SEP> difficile <SEP> I-E <SEP> t <SEP> 1.56 <SEP> 0.78 <SEP> - <SEP> - <SEP> 12.5 <tb> Table 1-2 In vitro antibacterial activity (standard strain) EMI35.1 <tb> <SEP> MIC(pg/ml) <tb> <SEP> Organism <SEP> (106 <SEP> cells/ml) <SEP> Gram <tb> <SEP> Exp.8 <SEP> Exp.9 <SEP> Exp.10 <SEP> Exp.ll <SEP> Exp.12 <tb> Bacillus <SEP> subtilis <SEP> PCI <SEP> 219 <SEP> + <SEP> 0.0125 <SEP> 0.05 <SEP> 0.0125 <SEP> 0.025 <SEP> 0.0125 <tb> Staphylococcus <SEP> aureus <SEP> 209 <SEP> P <SEP> + <SEP> 0.025 <SEP> 0.20 <SEP> 0.05 <SEP> 0.05 <SEP> 0.0125 <tb> S. <SEP> aureus <SEP> Smith <SEP> + <SEP> 0.025 <SEP> 0.39 <SEP> 0.05 <SEP> 0.05 <SEP> 0.025 <tb> S. <SEP> aureus <SEP> IID <SEP> 670 <SEP> (Terajima) <SEP> + <SEP> 0.025 <SEP> 0.20 <SEP> 0.05 <SEP> 0.05 <SEP> 0.025 <tb> S. <SEP> epidermidis <SEP> IID <SEP> 866 <SEP> + <SEP> 0.025 <SEP> 0.10 <SEP> 0.10 <SEP> 0.05 <SEP> 0.025 <tb> Streptococcus <SEP> pyogenes <SEP> S-8 <SEP> + <SEP> 0.05 <SEP> 0.20 <SEP> 0.39 <SEP> 0.20 <SEP> 0.05 <tb> S. <SEP> pyogenes <SEP> IID <SEP> 692 <SEP> + <SEP> 0.05 <SEP> 0.20 <SEP> 0.39 <SEP> 0.20 <SEP> 0.05 <tb> S. <SEP> pneumoniae <SEP> llD <SEP> 552 <SEP> + <SEP> 0.05 <SEP> 0.39 <SEP> 0.39 <SEP> 0.39 <SEP> 0.05 <tb> S. <SEP> faecalis <SEP> IID <SEP> 682 <SEP> t <SEP> <SEP> 0.10 <SEP> 0.39 <SEP> 0.39 <SEP> 0.39 <SEP> 0.05 <tb> Escherichia <SEP> coli <SEP> Mill <SEP> JC-2 <SEP> - <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.025 <tb> E. <SEP> coli <SEP> ATCC <SEP> 10536 <SEP> - <SEP> 0.025 <SEP> 0.10 <SEP> 0.10 <SEP> 0.05 <SEP> 0.025 <tb> E. <SEP> coli <SEP> ML <SEP> 4707 <SEP> - <SEP> 0.025 <SEP> 0.10 <SEP> 0.20 <SEP> 0.05 <SEP> 0.025 <tb> Proteus <SEP> vulgarin <SEP> IFO <SEP> 3167 <SEP> - <SEP> 0.025 <SEP> 0.10 <SEP> 0.025 <SEP> 0.05 <SEP> 0.025 <tb> P. <SEP> mirabilis <SEP> llD <SEP> 994 <SEP> - <SEP> 0.05 <SEP> 0.39 <SEP> 0.025 <SEP> 0.10 <SEP> 0.025 <tb> Morganella <SEP> mop <SEP> anti <SEP> lID <SEP> 602 <SEP> - <SEP> 0.20 <SEP> 0.78 <SEP> 0.025 <SEP> 0.39 <SEP> 0.10 <tb> Enterobacter <SEP> cloacae <SEP> IID <SEP> 977 <SEP> - <SEP> 0.10 <SEP> 0.39 <SEP> 0.20 <SEP> 0.39 <SEP> 0.05 <tb> Citrobacter <SEP> freundii <SEP> llD <SEP> 976 <SEP> - <SEP> 0.025 <SEP> 0.20 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <tb> Klebsiella <SEP> pneumoniae <SEP> KY(GN)6445 <SEP> - <SEP> 0.025 <SEP> 0.10 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <tb> K. <SEP> pneumoniae <SEP> 1-220S <SEP> - <SEP> 0.10 <SEP> 0.39 <SEP> 0.20 <SEP> 0.20 <SEP> 0.05 <tb> Salmonella <SEP> entritidis <SEP> llD <SEP> 604 <SEP> - <SEP> 0.05 <SEP> 0.20 <SEP> 0.10 <SEP> 0.20 <SEP> 0.05 <tb> Shigella <SEP> sonnei <SEP> IID <SEP> 969 <SEP> - <SEP> 0.0125 <SEP> 0.10 <SEP> 0.025 <SEP> 0.05 <SEP> ' <SEP> 0.025 <tb> Yersinia <SEP> enterocolitica <SEP> IID <SEP> 981 <SEP> - <SEP> 0.10 <SEP> 0.39 <SEP> 0.10 <SEP> 0.20 <SEP> 0.05 <tb> Sorratia <SEP> marcescens <SEP> IID <SEP> 618 <SEP> 0.20 <SEP> 0.39 <SEP> 0.20 <SEP> 0.39 <SEP> 0.10 <tb> S. <SEP> marcescens <SEP> GN <SEP> 7577 <SEP> - <SEP> 1.56 <SEP> 6.25 <SEP> 3.13 <SEP> 3.13 <SEP> 0.78 <tb> Pseudomonas <SEP> aeruginosa <SEP> V-1 <SEP> - <SEP> 0.78 <SEP> 1.56 <SEP> 1.56 <SEP> 1.56 <SEP> 0.39 <tb> P. <SEP> aeruginosa <SEP> IFO <SEP> 12689 <SEP> - <SEP> 1.56 <SEP> 3.13 <SEP> 3.13 <SEP> 3.13 <SEP> 0.78 <tb> P. <SEP> aeruginosa <SEP> IID <SEP> 1210 <SEP> - <SEP> 1.56 <SEP> 3.13 <SEP> 3.13 <SEP> 3.13 <SEP> 1.56 <tb> Acinetobacter <SEP> anitratus <SEP> llD <SEP> 876 <SEP> - <SEP> 0.10 <SEP> 0.78 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <tb> Alcaligenes <SEP> faecalis <SEP> 0104002 <SEP> - <SEP> 0.39 <SEP> 1.56 <SEP> 0.78 <SEP> 1.56 <SEP> 0.39 <tb> Becteroides <SEP> fragilis <SEP> GM <SEP> 7000 <SEP> - <SEP> 0.78 <SEP> 25 <SEP> 1.56 <SEP> 1.56 <SEP> 0.20 <tb> B. <SEP> fragilis <SEP> 0558 <SEP> - <SEP> 0.39 <SEP> 12.5 <SEP> 0.78 <SEP> 0.78 <SEP> 0.10 <tb> B. <SEP> fragilis <SEP> 25285 <SEP> - <SEP> 0.39 <SEP> 25 <SEP> 0.78 <SEP> 0.78 <SEP> 0.10 <tb> Fusobacterium <SEP> varium <SEP> KYA <SEP> 8501 <SEP> - <SEP> 0.78 <SEP> 25 <SEP> 6.25 <SEP> 3.13 <SEP> 0.39 <tb> Clostridium <SEP> Perfringens <SEP> KYA <SEP> 13123 <SEP> + <SEP> 0.39 <SEP> 1.56 <SEP> 0.39 <SEP> 0.20 <SEP> 0.10 <tb> C. <SEP> ramosum <SEP> t <SEP> 0.78 <SEP> 12.5 <SEP> 1.56 <SEP> 0.78 <SEP> 0.39 <tb> C. <SEP> difficile <SEP> I-E <SEP> + <SEP> 1.56 <SEP> 12.5 <SEP> 6.25 <SEP> 3.13 <SEP> - <SEP> <tb> Table 1-3 In vitro antibacterial activity (standard strain) EMI36.1 <tb> <SEP> MIC <SEP> ( g/ml) <tb> <SEP> Organism <SEP> (106 <SEP> cells/ml) <SEP> Gram <tb> <SEP> Exp.13 <SEP> Exp.14 <SEP> Exp.15 <SEP> Exp.16 <SEP> Exp.17 <tb> Bacillus <SEP> subtilis <SEP> PCI <SEP> 219 <SEP> + <SEP> 0.0125 <SEP> <SEP> û.0125 <SEP> 0.0125 <SEP> 0.0125 <SEP> 0.0125 <SEP> <tb> Staphylococcus <SEP> aureus <SEP> 209 <SEP> P <SEP> + <SEP> 0.0125 <SEP> 0.0125 <SEP> 0.0125 <SEP> 0.0125 <SEP> 0.0125 <tb> S. <SEP> aureus <SEP> Smith <SEP> t <SEP> <SEP> 0.025 <SEP> 0.0125 <SEP> 0.0125 <SEP> 0.0125 <SEP> 0.0125 <tb> S. <SEP> aureus <SEP> llD <SEP> 670 <SEP> (Terajima) <SEP> t <SEP> 0.025 <SEP> 0.0125 <SEP> 0.0125 <SEP> 0.025 <SEP> 0.025 <tb> s.epidermin <SEP> llp886 <SEP> + <SEP> 0.125 <SEP> 0.0125 <SEP> 0,0125 <SEP> 0,025 <tb> Streptococcus <SEP> pygenes <SEP> S-8 <SEP> + <SEP> 0.5 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <tb> S. <SEP> pyoqenes <SEP> IID <SEP> 692 <SEP> t <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <tb> S. <SEP> pneumoniae <SEP> IID <SEP> 552 <SEP> + <SEP> 0.05 <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <tb> S. <SEP> faecalis <SEP> llD <SEP> 682 <SEP> + <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.05 <tb> Escherichie <SEP> col2 <SEP> NIBJ <SEP> JC-2 <SEP> - <SEP> 0.0125 <SEP> 0.012560.0063 <SEP> 0.0125 <SEP> 00125 <tb> E. <SEP> coli <SEP> ATCC <SEP> 10536 <SEP> - <SEP> 0.025 <SEP> 0.0125 <SEP> 0.0125 <SEP> 0.025 <SEP> 0.025 <tb> E. <SEP> coli <SEP> ML <SEP> 4707 <SEP> - <SEP> 0.025 <SEP> 0.0125 <SEP> 0.025 <SEP> 0.025 <SEP> 0.025 <tb> Proteus <SEP> vulgeris <SEP> IFO <SEP> 3167 <SEP> - <SEP> 0.025 <SEP> 0.025 <SEP> 0.0125 <SEP> 0.025 <SEP> 0.025 <tb> P. <SEP> mirabilis <SEP> IID <SEP> 994 <SEP> - <SEP> 0.05 <SEP> 0.025 <SEP> 0.0125 <SEP> 0.025 <SEP> 0.025 <tb> Morganella <SEP> organic <SEP> IID <SEP> 602 <SEP> - <SEP> 0.10 <SEP> 0.05 <SEP> 0.10 <SEP> 0.10 <SEP> 0.20 <tb> Enterobacter <SEP> cloacae <SEP> IID <SEP> 977 <SEP> - <SEP> 0.10 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.10 <tb> Citrobacter <SEP> freundii <SEP> IID <SEP> 976 <SEP> - <SEP> 0.05 <SEP> 0.025 <SEP> 0.025 <SEP> 0.105 <SEP> 0.05 <tb> Klebsiella <SEP> pneumoniae <SEP> KY(GN)6445 <SEP> - <SEP> 0.05 <SEP> 0.025 <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <tb> K <SEP> , <SEP> neumoniee <SEP> 3-2208 <SEP> - <SEP> 0.18 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.10 <tb> Salmonella <SEP> entritidis <SEP> IID <SEP> 604 <SEP> - <SEP> 0.05 <SEP> 0.025 <SEP> 0.025 <SEP> 0.05 <SEP> 0.10 <tb> Shiella <SEP> sonnei <SEP> IID <SEP> 969 <SEP> - <SEP> 0.025 <SEP> 0.0125 <SEP> 0.0125 <SEP> 0.025 <SEP> 0.025 <tb> Yersinia <SEP> enterocolitics <SEP> llD <SEP> 981 <SEP> - <SEP> 0.10 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.10 <tb> Serratia <SEP> marcescens <SEP> llD <SEP> 618 <SEP> - <SEP> 0.10 <SEP> 0.05 <SEP> 0.10 <SEP> 0.10 <SEP> 0.10 <tb> S. <SEP> marcescens <SEP> GN <SEP> 7577 <SEP> - <SEP> 0.78 <SEP> 0.39 <SEP> 1 <SEP> 0.78 <SEP> 0.78 <SEP> 0.78 <tb> Pseudomoncs <SEP> seruginosa <SEP> V-1- <SEP> - <SEP> o.78 <SEP> 0.20 <SEP> 0.10 <SEP> 1.20 <SEP> 0.35 <tb> P. <SEP> aerugínosa <SEP> IFO <SEP> 12689 <SEP> - <SEP> 1.56 <SEP> 0.39 <SEP> ' <SEP> 0.78 <SEP> 1.56 <SEP> 3.13 <tb> P. <SEP> aeruginosa <SEP> IID <SEP> 1210 <SEP> - <SEP> 1.56 <SEP> 0.39 <SEP> 0.78 <SEP> 1.56 <SEP> 1.56 <tb> Acinetobacter <SEP> anitratus <SEP> lld <SEP> 876 <SEP> - <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <tb> Alcaligenes <SEP> faecalin <SEP> 0104002 <SEP> - <SEP> 0.39 <SEP> 0.20 <SEP> 0.20 <SEP> 0.39 <SEP> 0.39 <tb> Bacteroides <SEP> fragilis <SEP> GM <SEP> 7000 <SEP> - <SEP> 0.10 <SEP> 0.10 <SEP> 0.10 <SEP> 0.20 <SEP> 0.10 <tb> B. <SEP> fragilis <SEP> 0558 <SEP> - <SEP> 0.10 <SEP> 0.05 <SEP> 10.05 <SEP> 0.10 <SEP> < 0.05 <tb> B. <SEP> fragilis <SEP> 25285 <SEP> - <SEP> 0.05 <SEP> 0.05 <SEP> 10.05 <SEP> 0.10 <SEP> < 0.05 <tb> Fusobacterium <SEP> varium <SEP> KYA <SEP> 8501 <SEP> - <SEP> - <SEP> 0.39 <SEP> 0.39 <SEP> 0.39 <SEP> 0.39 <SEP> 0.39 <tb> Clostridium <SEP> perfringens <SEP> KYA <SEP> 13123 <SEP> + <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.10 <SEP> 0.10 <tb> C. <SEP> ramosum <SEP> t <SEP> 0.20 <SEP> 0.39 <SEP> 0.39 <SEP> 0.39 <SEP> 0.39 <tb> C. <SEP> difficile <SEP> I-E <SEP> + <SEP> - <SEP> - <SEP> - <SEP> - <SEP> - <SEP> <tb> Table 1-4 In vitro antibacterial activity (standart strain) EMI37.1 <tb> <SEP> MIC <SEP> ( g/ml) <tb> <SEP> Organism <SEP> (106 <SEP> cells/ml) <SEP> Gram <tb> <SEP> Exp.20 <SEP> Exp.21 <SEP> Ref.l <SEP> CFLX <SEP> NFLX <tb> Bacillus <SEP> subtilis <SEP> PCI <SEP> 219 <SEP> + <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <tb> Staphylococcus <SEP> aureus <SEP> 209 <SEP> P <SEP> + <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <SEP> 0.20 <SEP> 0.78 <tb> S. <SEP> aureus <SEP> Smith <SEP> t <SEP> <SEP> 0.025 <SEP> 0.05 <SEP> 0.10 <SEP> 0.39 <SEP> 1.56 <tb> S. <SEP> aureus <SEP> lID <SEP> 670 <SEP> Tetrajima <SEP> + <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.78 <tb> S <SEP> Epdermidin <SEP> llD <SEP> + <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <tb> Streptococcus <SEP> pyogenes <SEP> S-8 <SEP> t <SEP> <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.78 <SEP> 1.56 <tb> S <SEP> pyogenes <SEP> llD <SEP> 692 <SEP> + <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <SEP> 0.78 <SEP> 3.13 <tb> S. <SEP> opeu@omnse <SEP> llD <SEP> 552 <SEP> t <SEP> <SEP> 0.05 <SEP> 0.20 <SEP> 0.20 <SEP> 0.78 <SEP> 3.13 <tb> S. <SEP> faecalis <SEP> IID <SEP> 682 <SEP> + <SEP> 0.10 <SEP> 0.20 <SEP> 0.20 <SEP> 0.78 <SEP> 1.56 <tb> Escherichis <SEP> coli <SEP> NIHJ <SEP> JC-2 <SEP> - <SEP> 0.025 <SEP> 0.05 <SEP> 0.10 <SEP> 0.0125 <SEP> 0.025 <tb> E. <SEP> coli <SEP> ATCC <SEP> 10536 <SEP> - <SEP> 0.025 <SEP> 0.05 <SEP> 0.10 <SEP> 0.025 <SEP> 0.05 <tb> E. <SEP> coli <SEP> ML <SEP> 4707 <SEP> - <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.0125 <SEP> 0.05 <tb> Proteus <SEP> vulqaris <SEP> IFO <SEP> 3167 <SEP> - <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.0125 <SEP> 0.025 <tb> P. <SEP> mirabilin <SEP> IID <SEP> 994 <SEP> - <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.025 <SEP> 0.05 <tb> Morganella <SEP> mop <SEP> anti <SEP> llD <SEP> 602 <SEP> - <SEP> 0.20 <SEP> 0.20 <SEP> 0.78 <SEP> 0.05 <SEP> 0.05 <tb> Enterobacter <SEP> cloacas <SEP> IID <SEP> 977 <SEP> - <SEP> 0.10 <SEP> 0.20 <SEP> 0.20 <SEP> 0.05 <SEP> 0.10 <tb> Citrobacter <SEP> freundii <SEP> IID <SEP> 976 <SEP> - <SEP> 0.05 <SEP> 0.05 <SEP> 0.20 <SEP> 0.025 <SEP> 0.05 <tb> Klebssielle <SEP> pneumonim <SEP> KY(GN)6645 <SEP> - <SEP> 0,05 <SEP> 0.20 <SEP> 0.025 <SEP> 0.05 <tb> K. <SEP> pneuoniae <SEP> 1-2205 <SEP> - <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <SEP> 0.05 <SEP> 0.20 <tb> Salmonella <SEP> entritidis <SEP> IID <SEP> 604 <SEP> - <SEP> 0.10 <SEP> 0.10 <SEP> 0.39 <SEP> 0.05 <SEP> 0.10 <tb> Shigelle <SEP> sonnei <SEP> IID <SEP> 969 <SEP> - <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.0125 <SEP> 0.05 <tb> Yersinia <SEP> enterocolitica <SEP> IID <SEP> 981 <SEP> - <SEP> 0.10 <SEP> 0.10 <SEP> 0.39 <SEP> 0.05 <SEP> 0.10 <tb> Serratia <SEP> marcescens <SEP> IID <SEP> 618 <SEP> - <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <SEP> 0.05 <SEP> 0.05 <tb> S. <SEP> macar*cens <SEP> GN <SEP> 7577 <SEP> - <SEP> 1.56 <SEP> 1.56 <SEP> 6.25 <SEP> 0.78 <SEP> 1.56 <tb> Pseudosotias <SEP> aeruginosa <SEP> V-1 <SEP> - <SEP> 0.39 <SEP> 0.39 <SEP> 6.25 <SEP> 0.39 <SEP> 0.78 <tb> P. <SEP> aeru:inosa <SEP> IFO <SEP> 12689 <SEP> - <SEP> 3.13 <SEP> 3.13 <SEP> 6.25 <SEP> 0.39 <SEP> 0.78 <tb> P. <SEP> serugina <SEP> IID <SEP> 1210 <SEP> - <SEP> 1.56 <SEP> 3.13 <SEP> 12.5 <SEP> 1.56 <SEP> 3.13 <tb> Acinetobacter <SEP> anitratus <SEP> lID <SEP> 876 <SEP> - <SEP> 0.05 <SEP> 0.05 <SEP> 0.78 <SEP> 0.39 <SEP> 3.13 <tb> Alcaligenes <SEP> faecalia <SEP> 0104002 <SEP> - <SEP> 0.39 <SEP> 0.78 <SEP> 12.5 <SEP> 0.39 <SEP> 3.13 <tb> Bacteroides <SEP> fragilis <SEP> GM <SEP> 7000 <SEP> - <SEP> 0.10 <SEP> 0.39 <SEP> 12.5 <SEP> 3.13 <SEP> 25 <tb> B, <SEP> fragilis <SEP> 0558 <SEP> - <SEP> 0.05 <SEP> 0.20 <SEP> 0,25 <SEP> 3.23 <SEP> 25 <tb> B. <SEP> fra <SEP> ilis <SEP> 25285 <SEP> - <SEP> =0.05 <SEP> 0.20 <SEP> 6.25 <SEP> 3.13 <SEP> 25 <tb> Fusobacterium <SEP> varium <SEP> KYA8501 <SEP> - <SEP> 0.78 <SEP> 3.13 <SEP> 6.25 <SEP> 17.5 <SEP> 100 <tb> Clostridium <SEP> perfringens <SEP> KYA <SEP> 13123 <SEP> + <SEP> 0.10 <SEP> 0.20 <SEP> 1.56 <SEP> 0.39 <SEP> 1.56 <tb> C. <SEP> ramosum <SEP> + <SEP> 0.39 <SEP> 0.78 <SEP> 6.25 <SEP> 3.13 <SEP> 50 <tb> c.difficile <SEP> I-E <SEP> + <SEP> - <SEP> - <SEP> - <SEP> 12.5 <SEP> 50 <tb> Ref.1; 1-Ethyl-7-(3-ethylaminoethyl-1-pyrrlidinyl)-6,8-difluoro-1,4- dihydro-4 oxo-3-quinolinecarboxylic acid CFI.X: Ciprofloxacin NFLX: Norfioxacin Table 2-1 In vitro antibacterial activity(1) (clinical isolates) EMI38.1 Organism <SEP> Mic( g/ml) <tb> 106 <SEP> cells/ml) <SEP> Gram <SEP> Exp.1 <SEP> Exp.2 <SEP> Exp.4 <SEP> Exp.b <SEP> Exp.7 <SEP> Exp.8 <SEP> Exp.9 <SEP> Exp.10 <SEP> Exp.11 <SEP> Exp.12 <SEP> Exp.13 <SEP> Exp.14 <SEP> Exp.15 <SEP> Exp.16 <tb> S.pneumoniae <SEP> 15 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.20 <SEP> 0.39 <SEP> 0.20 <SEP> 0.39 <SEP> 0.39 <SEP> 0.20 <SEP> 0.05 <SEP> 0.025 <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <tb> S.pneumoniae <SEP> 24 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> - <SEP> - <SEP> 0.05 <SEP> - <SEP> - <SEP> - <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> - <SEP> S.pneumoniae <SEP> 28 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> - <SEP> - <SEP> 0.05 <SEP> - <SEP> - <SEP> - <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> - <SEP> S.pneumoniae <SEP> 2054 <SEP> + <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.39 <SEP> 0.39 <SEP> 0.05 <SEP> 0.78 <SEP> 0.78 <SEP> 0.20 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <tb> S.pneumoniae <SEP> 2950 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.20 <SEP> 0.39 <SEP> 0.10 <SEP> 0.78 <SEP> 0.39 <SEP> 0.20 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <tb> S.pneumoniae <SEP> 3227 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.20 <SEP> 0.39 <SEP> - <SEP> 0.20 <SEP> 0.39 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <tb> S.pyogenes <SEP> 3130 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.20 <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <tb> S.pyogenes <SEP> 3102 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.20 <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.20 <SEP> 0.05 <SEP> 0.05 <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <tb> S.pyogenes <SEP> 3107 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.20 <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.20 <SEP> 0.05 <SEP> 0.05 <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <tb> S.pyogenes <SEP> 4340 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.05 <SEP> 0.20 <SEP> 0.05 <SEP> 0.20 <SEP> 0.20 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <tb> S.pyogenes <SEP> 4372 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.05 <SEP> 0.20 <SEP> 0.39 <SEP> 0.20 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <tb> S.agalactiae <SEP> 4394 <SEP> 0.10 <SEP> 0.20 <SEP> 0.10 <SEP> 0.39 <SEP> 0.39 <SEP> 0.10 <SEP> 0.78 <SEP> 0.78 <SEP> 0.39 <SEP> 0.10 <SEP> 0.10 <SEP> 0.10 <SEP> 0.20 <SEP> 0.10 <tb> S.agalactiae <SEP> 4049 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.39 <SEP> 0.78 <SEP> 0.20 <SEP> 0.78 <SEP> 0.78 <SEP> 0.39 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <tb> S.agalactiae <SEP> 4342 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.20 <SEP> 0.39 <SEP> 0.20 <SEP> 0.39 <SEP> 0.20 <SEP> 0.20 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <tb> S.agalactiae <SEP> 4470 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.20 <SEP> 0.39 <SEP> - <SEP> 0.39 <SEP> 0.39 <SEP> 0.20 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <tb> S.agalactiae <SEP> 4368 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.20 <SEP> 0.39 <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <SEP> 0.20 <SEP> 0.05 <SEP> 0.025 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <tb> S.agalactiae <SEP> 4468 <SEP> 0.05 <SEP> 0.10 <SEP> 0.05 <SEP> 0.20 <SEP> 0.39 <SEP> 0.10 <SEP> 0.39 <SEP> 0.39 <SEP> 0.20 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <tb> S.fecalis <SEP> 49 <SEP> 0.10 <SEP> 0.20 <SEP> 0.10 <SEP> 0.39 <SEP> 0.78 <SEP> 0.10 <SEP> 0.39 <SEP> 0.39 <SEP> 0.20 <SEP> 0.05 <SEP> 0.05 <SEP> 0.05 <SEP> 0.20 <SEP> 0.10 <tb> S.fecalis <SEP> 214 <SEP> 0.10 <SEP> 0.20 <SEP> 0.10 <SEP> 0.39 <SEP> 0.78 <SEP> 0.05 <SEP> 0.39 <SEP> 0.39 <SEP> 0.39 <SEP> 0.10 <SEP> 0.10 <SEP> 0.05 <SEP> 0.20 <SEP> 0.10 <tb> S.fecalis <SEP> 401 <SEP> 0.10 <SEP> 0.20 <SEP> 0.10 <SEP> 0.39 <SEP> 0.78 <SEP> 0.10 <SEP> 0.78 <SEP> 0.78 <SEP> 0.39 <SEP> 0.10 <SEP> 0.10 <SEP> 0.05 <SEP> 0.20 <SEP> 0.10 <tb> S.fecalis <SEP> 402 <SEP> 0.10 <SEP> 0.20 <SEP> 0.10 <SEP> 0.39 <SEP> 0.78 <SEP> 0.20 <SEP> 0.78 <SEP> 0.39 <SEP> 0.39 <SEP> 0.10 <SEP> 0.10 <SEP> 0.05 <SEP> 0.20 <SEP> 0.10 <tb> Table 2-2 In vitro antibacterial activity (2) (clinical isolates) EMI39.1 <tb> <SEP> Organism <SEP> MIC( g/ml) <tb> <SEP> (106 <SEP> cells/ml <SEP> Gram <SEP> Exp.17 <SEP> Exp.20 <SEP> Exp.21 <SEP> Ref.1 <SEP> CFLX <SEP> NFLX <tb> S. <SEP> pneumoniae <SEP> 15 <SEP> O.05 <SEP> O.05 <SEP> <SEP> O.20 <SEP> <SEP> O.20 <SEP> <SEP> O.78 <SEP> <SEP> 3.13 <tb> S. <SEP> pneumoniae <SEP> 24 <SEP> - <SEP> - <SEP> - <SEP> - <SEP> 1.56 <SEP> 12.5 <tb> S. <SEP> pneumoniae <SEP> 28 <SEP> - <SEP> - <SEP> - <SEP> - <SEP> 0.78 <SEP> 6.25 <SEP> <tb> S. <SEP> pneumoniae <SEP> 2054 <SEP> + <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <SEP> 1.56 <SEP> 6.25 <tb> S. <SEP> pneumoniae <SEP> 2950 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.39 <SEP> 1.56 <SEP> 12.5 <tb> S. <SEP> pneumoniae <SEP> 3227 <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <SEP> 1.56 <SEP> 12.5 <tb> S. <SEP> pyogenes <SEP> 3130 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <SEP> 1.56 <tb> S. <SEP> pyogenes <SEP> 3102 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <SEP> 1.56 <tb> S. <SEP> pyogenes <SEP> 3107 <SEP> + <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.10 <SEP> 0.39 <SEP> 1.56 <tb> S.pyogenes <SEP> 4340 <SEP> 0.05 <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.78 <SEP> 1.56 <tb> S.pyogenes <SEP> 4372 <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.20 <SEP> 0.78 <SEP> 1.56 <tb> S.agalactiae <SEP> 4394 <SEP> 0.10 <SEP> 0.20 <SEP> 0.20 <SEP> 0.78 <SEP> 3.13 <SEP> 6.25 <tb> S.agalactiae <SEP> 4049 <SEP> 0.05 <SEP> 0.15 <SEP> 0.20 <SEP> 0.39 <SEP> 0.78 <SEP> 6.25 <tb> S.agalactiae <SEP> 4342 <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <SEP> 0.78 <SEP> 6.25 <tb> S. <SEP> agalactiae <SEP> 4470 <SEP> + <SEP> 0.05 <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <SEP> 0.78 <SEP> 6.25 <tb> S. <SEP> agalactiae <SEP> 4368 <SEP> 0.05 <SEP> 0.05 <SEP> 0.20 <SEP> 0.20 <SEP> 0.78 <SEP> 3.13 <tb> S. <SEP> agalactiae <SEP> 4468 <SEP> 0.05 <SEP> 0.10 <SEP> 0.120 <SEP> 0.39 <SEP> 0.78 <SEP> 3.13 <tb> S. <SEP> faecalis <SEP> 49 <SEP> 0.10 <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <SEP> 1.56 <SEP> 3.13 <tb> S. <SEP> faecalis <SEP> 214 <SEP> 0.10 <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <SEP> 1.56 <SEP> 3.13 <tb> S. <SEP> faecalis <SEP> 401 <SEP> + <SEP> 0.10 <SEP> 0.10 <SEP> 0.20 <SEP> 0.39 <SEP> 1.56 <SEP> 6.25 <tb> S. <SEP> faecalis <SEP> 402 <SEP> 0.10 <SEP> 0.10 <SEP> 0.20 <SEP> 0.78 <SEP> 1.56 <SEP> 6.25 <tb> Experiment-2 In vivo antibacterial activity against systemic infection in mice (ICR-S) The infections were produced by intraperitoneal injection of a suspension of the bacterial culture corresponding to the germ studied. The products were administered orally at an hour after the infection. The 50% effective dose (ED50) which protects 50 % of the animals from death caused by the infection was determined from the relation between dosage and survival rate. The efficacy of the compound of this invention was shown in Table 3 together with that of the known compound. The present compounds were the most effective against grampositive bacterial infections and, amongest these, those of Example 2, 13, 17 and 21 had also greater activity against gramnegative bacterial infections than the reference compounds. Table 3-1 In vivo antibacterial activity against systemic infection in mice (ICR-S) EMI40.1 <tb> <SEP> MIC <SEP> ED50 <SEP> (mg/kg) <SEP> against <SEP> S.aureus <SEP> Smith <tb> Compound <SEP> (Ug/ml) <SEP> Challenge <SEP> dose <SEP> (cfu/mouse) <tb> <SEP> 7 <SEP> x <SEP> i05 <SEP> 5.5 <SEP> x <SEP> i05 <SEP> <tb> Example <SEP> 1 <SEP> 0.05 <SEP> 0.9 <tb> Example <SEP> 2 <SEP> 0.025 <SEP> 0.97 <tb> Example <SEP> 4 <SEP> 0.05 <SEP> 1.57 <tb> <SEP> CFLX <SEP> i <SEP> 0.20 <SEP> 12.3 <SEP> 25.5 <tb> (n=5) Table 3-2 In vivo antibacterial activity against systemic infection in mice (ICR-S) ED50 (mg/kg) EMI41.1 <tb> Compound <SEP> S. <SEP> aureus <SEP> S. <SEP> pneumoniae <SEP> E. <SEP> coli <tb> <SEP> Smitha) <SEP> S-4288b) <SEP> ML4707c) <tb> Example <SEP> 2 <SEP> 0.9 <SEP> 4.6 <SEP> 0.8 <tb> <SEP> 4 <SEP> 1.8 <SEP> 13.8 <SEP> 3.0 <tb> <SEP> 8 <SEP> 1.1 <SEP> 16.1 <SEP> 1.9 <tb> <SEP> 12 <SEP> 0.5 <SEP> 6.5 <SEP> 1.9 <tb> <SEP> 13 <SEP> 0.6 <SEP> 2.0 <SEP> 0.7 <tb> <SEP> 14 <SEP> - <SEP> 10.0 <SEP> 5.4 <tb> <SEP> 15 <SEP> - <SEP> - <SEP> 7.1 <tb> <SEP> 16 <SEP> - <SEP> - <SEP> 1.9 <tb> <SEP> 17 <SEP> - <SEP> - <SEP> 0.9 <tb> <SEP> 20 <SEP> - <SEP> - <SEP> 2.1 <tb> <SEP> 21 <SEP> - <SEP> - <SEP> 1.2 <tb> <SEP> Ref.1 <SEP> 5.0 <SEP> 62.2 <SEP> > 10 <tb> <SEP> CF <SEP> > 25 <SEP> > 100 <SEP> 1.8 <tb> a) Challenge dose s 6.0 x 105 cfu/mouse b) Challenge dose s 7.5 x 104 - 4.8 x 106 cfu/mouse c) Challenge dose : 2.7 x 106 - 8.8 x 106 cfu/mouse Experiment-3 In vitro antibacterial activity against Mycoplasma The antimicrobial activity of the present compounds has been investigated in vitro on M. pneumoniae Mac. Table 4 gives the minimum inhibitory concentrations (MIC) of the present compounds together with those of the reference compounds. The present compounds were the most active. Table 4 In vitro antibacterial activity against Mycoplasma (M. pneumoniae Mac.) EMI42.1 <tb> Compound <SEP> MIC* <SEP> ( <SEP> pg/ml <SEP> ) <SEP> <tb> Example <SEP> 2 <SEP> 0.05 <SEP> <tb> <SEP> 4 <SEP> 0.025 <tb> <SEP> 8 <SEP> 0.05 <tb> <SEP> 12 <SEP> 0.05 <tb> <SEP> 13 <SEP> 0.05 <tb> <SEP> Ref.l <SEP> 0.39 <tb> <SEP> CFLX <SEP> 0.78 <tb> <SEP> NFLX <SEP> 3.13 <tb> <SEP> TC <SEP> 0.78 <tb> TC : Tetracycline *The MIC was defined as the concentration of agent at which there was no visible growth after incubation for 5 days. Experiment-4 Absorption, distribution and excretion The blood, organs, bile and urine of the fasted rats, which had received the dose of 10 mg/kg of the test compound by oral administration, were sampled over a period of time and the pro portion of active compound was determined bacteriologically by the cup method using E. coli NIHJ JC-2. The results were shown in Table 5 and 6. All those results make it possible to guarantee a greater therapeutic action of the present compound than that of the reference compound because of higher concentration of active compound in comparition with the reference compound. Table 5 Concentration in organs ( g/ml or g) EMI44.1 <SEP> Time <SEP> after <SEP> administration <SEP> (hr) <tb> Organ <SEP> Compound <tb> <SEP> 1/2 <SEP> 1 <SEP> 2 <SEP> 4 <SEP> 6 <SEP> 8 <tb> Serum <SEP> Example <SEP> 2 <SEP> 1.47 <SEP> # <SEP> 0.19 <SEP> 1.30 <SEP> # <SEP> 0.20 <SEP> 0.90 <SEP> # <SEP> 0.19 <SEP> 0.42 <SEP> # <SEP> 0.15 <SEP> 0.26 <SEP> # <SEP> 0.09 <SEP> 0.12 <SEP> # <SEP> 0.04 <tb> <SEP> 13 <SEP> 1.72 <SEP> # <SEP> 0.37 <SEP> 1.12 <SEP> # <SEP> 0.30 <SEP> 0.57 <SEP> # <SEP> 0.08 <SEP> 0.23 <SEP> # <SEP> 0.04 <SEP> 0.10 <SEP> # <SEP> 0.01 <SEP> 0.14 <SEP> # <SEP> 0.08 <tb> <SEP> CFLX <SEP> 0.47 <SEP> # <SEP> 0.28 <SEP> 1.38 <SEP> # <SEP> 0.11 <SEP> 0.25 <SEP> # <SEP> 0.05 <SEP> 0.09 <SEP> # <SEP> 0.03 <SEP> 0.05 <SEP> # <SEP> 0.01 <SEP> 0.03 <SEP> # <SEP> 0.01 <tb> Lung <SEP> Example <SEP> 2 <SEP> 3.84 <SEP> 0.42 <SEP> 2.94 <SEP> # <SEP> 0.22 <SEP> 1.92 <SEP> # <SEP> 0.33 <SEP> 1.12 <SEP> # <SEP> 0.31 <SEP> 0.78 <SEP> # <SEP> 0.19 <SEP> 0.38 <SEP> # <SEP> 0.08 <tb> <SEP> 13 <SEP> 3.17 <SEP> # <SEP> 0.71 <SEP> 2.66 <SEP> # <SEP> 0.42 <SEP> 1.54 <SEP> # <SEP> 0.21 <SEP> 0.70 <SEP> # <SEP> 0.08 <SEP> 0.42 <SEP> # <SEP> 0.05 <SEP> 0.45 <SEP> # <SEP> 0.15 <tb> <SEP> CFLX <SEP> 0.86 <SEP> # <SEP> 0.58 <SEP> 0.60 <SEP> # <SEP> 0.19 <SEP> 0.29 <SEP> # <SEP> 0.02 <SEP> 0.14 <SEP> # <SEP> 0.02 <SEP> 0.07 <SEP> # <SEP> 0.03 <SEP> 0.06 <SEP> # <SEP> 0.01 <tb> Liver <SEP> Example <SEP> 2 <SEP> 9.48 <SEP> # <SEP> 0.44 <SEP> 6.24 <SEP> # <SEP> 0.53 <SEP> 4.32 <SEP> # <SEP> 0.43 <SEP> 2.36 <SEP> # <SEP> 0.48 <SEP> 1.62 <SEP> # <SEP> 0.29 <SEP> 0.82 <SEP> # <SEP> 0.19 <tb> <SEP> 13 <SEP> 1.79 <SEP> # <SEP> 0.91 <SEP> 5.71 <SEP> # <SEP> 0.63 <SEP> 3.14 <SEP> # <SEP> 0.36 <SEP> 1.34 <SEP> # <SEP> 0.12 <SEP> 0.86 <SEP> # <SEP> 0.15 <SEP> 1.13 <SEP> # <SEP> 0.52 <tb> <SEP> CFLX <SEP> 5.17 <SEP> # <SEP> 2.59 <SEP> 3.28 <SEP> # <SEP> 0.98 <SEP> 1.49 <SEP> # <SEP> 0.37 <SEP> 0.39 <SEP> # <SEP> 0.05 <SEP> 0.21 <SEP> # <SEP> 0.11 <SEP> 0.15 <SEP> # <SEP> 0.05 <tb> Kidney <SEP> Example <SEP> 2 <SEP> 8.50 <SEP> # <SEP> 0.31 <SEP> 6.72 <SEP> # <SEP> 0.54 <SEP> 4.66 <SEP> # <SEP> 0.30 <SEP> 2.62 <SEP> # <SEP> 0.59 <SEP> 1.86 <SEP> # <SEP> 0.11 <SEP> 0.98 <SEP> # <SEP> 0.22 <tb> <SEP> 13 <SEP> 5.63 <SEP> # <SEP> 0.64 <SEP> 4.70 <SEP> # <SEP> 0.60 <SEP> 2.81 <SEP> # <SEP> 0.25 <SEP> 1.47 <SEP> # <SEP> 0.25 <SEP> 0.84 <SEP> # <SEP> 0.09 <SEP> 0.89 <SEP> # <SEP> 0.40 <tb> <SEP> CFLX <SEP> 3.58 <SEP> # <SEP> 2.61 <SEP> 2.05 <SEP> # <SEP> 0.64 <SEP> 1.12 <SEP> # <SEP> 0.34 <SEP> 0.21 <SEP> # <SEP> 0.12 <SEP> 0.10 <SEP> # <SEP> 0.04 <SEP> 0.06 <SEP> # <SEP> 0.02 <tb> Mean -S.d.(n=5) Table 6 Urinary and bile excretion EMI45.1 <SEP> cocentration( g/ml) <tb> Sample <SEP> compound <tb> <SEP> 0-3 <SEP> 3-6 <SEP> 6-24 <SEP> hr <SEP> 0-3 <SEP> 0-6 <SEP> 0-24 <SEP> hr <tb> Urine <SEP> Example <SEP> 2 <SEP> 44.2 <SEP> # <SEP> 27.9 <SEP> 114.8 <SEP> # <SEP> 61.0 <SEP> 22.1 <SEP> # <SEP> 9.8 <SEP> 3.5 <SEP> # <SEP> 2.0 <SEP> 6.7 <SEP> # <SEP> 4.3 <SEP> 13.5 <SEP> # <SEP> 4.6 <tb> <SEP> 13 <SEP> 36.1 <SEP> # <SEP> 9.2 <SEP> 39.4 <SEP> # <SEP> 13.3 <SEP> 7.6 <SEP> # <SEP> 3.6 <SEP> 2.7 <SEP> # <SEP> 2.1 <SEP> 4.8 <SEP> # <SEP> 1.8 <SEP> 7.4 <SEP> # <SEP> 1.5 <tb> <SEP> CFLX <SEP> 101 <SEP> # <SEP> 36.9 <SEP> 57. <SEP> # <SEP> 19. <SEP> 12. <SEP> # <SEP> 6 <SEP> 3.9 <SEP> # <SEP> 2.9 <SEP> 6.6 <SEP> # <SEP> 2.2 <SEP> 8.9 <SEP> # <SEP> 1'.8 <tb> Blile <SEP> Example <SEP> 2 <SEP> 17.1 <SEP> # <SEP> 10.0 <SEP> 16.1 <SEP> # <SEP> 6.2 <SEP> 10.1 <SEP> # <SEP> 2.3 <SEP> 2.4 <SEP> # <SEP> 1.6 <SEP> 4.3 <SEP> # <SEP> 2.1 <SEP> 9.7 <SEP> # <SEP> 3.1 <tb> <SEP> 13 <SEP> 36.4 <SEP> # <SEP> 9.9 <SEP> 19.7 <SEP> # <SEP> 8.0 <SEP> 11.3 <SEP> # <SEP> 8.3 <SEP> 4.9 <SEP> # <SEP> 1.5 <SEP> 6.9 <SEP> # <SEP> 1.4 <SEP> 12.9 <SEP> # <SEP> 4.9 <tb> <SEP> CFLX <SEP> 2 <SEP> 11.1 <SEP> # <SEP> 4.4 <SEP> 5.9 <SEP> # <SEP> 2.6 <SEP> 1.5 <SEP> # <SEP> 0.6 <SEP> 1.3 <SEP> # <SEP> 0.7 <SEP> 1.9 <SEP> # <SEP> 0.5 <SEP> 2.5 <SEP> # <SEP> 0.8 <tb> Mean # S.D.(n=5l)

Claims: What is claimed is 1. A compound of the formula (I) EMI46.1 wherein R, R1 and R2 are each independentlyhydrogen atom or lower alkyl group and Y is hydrogen atom or halogen atom; the hydrates or the pharmaceutically acceptable acid addition or alkali salts thereof. 2. A process for the preparation of a compound of the forrula (I) EMI46.2 wherein R, R1 and R2 are each independently hydrogen atom or lower alkyl group and Y is hydrogen atom or halogen atom; the hydrates or the pharmaceutically acceptable acid addition or alkali salts thereof, which comprises condensing a compound of the formula (II) EMI46.3 wherein R is hydrogen atom or lower alkyl group, X is halogen atom and Y is hydrogen atom or halogen atom; with a secondary amine of the formula (III) EMI47.1 wherein R1 and R2 are each independently hydrogen atom or lower alkyl group. 3. A process for the preparation of a compound of the formula (I) EMI47.2 wherein R is a hydrogen atom and R1 and R2 are each independently hydrogen atom or lower alkyl group and Y is hydrogen atom or halogen atom; the hydrates or the pharmaceutically acceptable acid addition or alkali salts thereof, which comprises hydrolyzing a compound of the formula (IV) EMI47.3 wherein R1 and R2 are each independently hydrogen atom or lower alkyl group, Y is hydrogen atom or halogen atom and A is lower alkyl group. 4. An antibacterial pharmaceutical composition comprising at least one compound according to claim 1 and an inert phar maceutically acceptable carrier.

Inventors: HIRAI, KEIJI, IRIKURA, TSUTOMU, ISHIZAKI, TAKAYOSHI, SUZUE, SEIGO
Applicants: KYORIN PHARMACEUTICAL CO., LTD.
Application year: 1985
Main classification: C07D401
IPC codes: C07D401, C07C63, A61P31, A61K31, C07D215