Quinolone compounds and antiviral application thereof

Abstract

The invention relates to the field of medicines, and particularly discloses quinolone compounds and antiviral application thereof. The quinolone compound provided by the invention has a structural formula as shown in a general formula 1. The quinolone compound disclosed by the invention has better inhibition on DNA (deoxyribonucleic acid) such as PRV (porcine reproductive and respiratory syndrome virus) and ASFV (anaerobic swine fever virus), has excellent ASFV and PRV resisting activity, and can be used for preparing ASFV and PRV resisting drugs. # imgabs0 #

Description

Technical Field

[0001] The present application relates to the field of medicine, specifically to a class of novel quinolone compounds and their antiviral uses, and more specifically to the application of a class of novel quinolone compounds in the field of anti-pseudorabies virus and African swine fever virus. Background Art

[0002] Pseudorabies virus (PRV) is a DNA virus that mainly infects pigs as its natural host. It can cause an acute and severe infectious disease characterized by fever and encephalomyelitis. The mortality rate of piglets infected with PRV is extremely high, up to 100%. African swine fever virus (ASFV) is an ancient virus that was first discovered in Kenya, Africa as early as 1921, and has a history of about 100 years. African swine fever is an acute, febrile, highly contagious animal disease of domestic pigs and wild boars caused by African swine fever virus. Pigs of all breeds and ages can be infected, with an incidence and mortality rate of up to 100%. The World Organization for Animal Health lists it as a notifiable animal disease, and China also lists it as a Class I animal disease. ASFV can enter the pig's body through the mouth and upper respiratory system, infect the nasopharynx or tonsils, and the virus quickly spreads to the mandibular lymph nodes and spreads throughout the body through lymph and blood. During a strong infection, cells change very quickly, and the cells die before showing an obvious stimulation response. When the virus is attenuated, the stimulus response is easily observed, the nucleus becomes larger, and mitosis generally occurs. The morbidity rate is usually between 40% and 85%, and the mortality rate varies depending on the infected strain. The mortality rate of highly pathogenic strains can be as high as 90%-100%; the mortality rate of moderately pathogenic strains in adult animals is between 20%-40%, and the mortality rate in young animals is between 70%-80%; the mortality rate of low pathogenic strains is between 10%-30%. At present, there are no reports of highly active drugs against DNA viruses such as PRV and ASFV, which cannot meet the needs of pig production. Summary of the invention

[0003] The purpose of the present invention is to provide a class of quinolone compounds and their antiviral applications in view of the deficiencies of the prior art. The quinolone compounds provided by the present invention have good inhibition on DNA such as PRV and ASFV, and can be used to prepare anti-ASFV and PRV virus drugs.

[0004] The present invention provides a quinolone compound described in general formula I, wherein the general formula I is:

[0005]

[0006] Wherein R1 is selected from Wherein Z is selected from an alkyl chain consisting of 1 to 4 carbon atoms, -NH-, -N(CH3)-, -CO-, -SO2-, and Y is selected from The above aromatic ring structures (including five-membered aromatic rings and six-membered aromatic rings) and the above aromatic ring structures substituted by -OH, halogen, CH3, CF3, NH2, alkoxy and other groups;

[0007] X is selected from N, O, when X is O, R2 is absent, when X is N, R2 is selected from H, alkyl, substituted alkyl, trifluoromethyl;

[0008] R3 is selected from alkyl, cycloalkyl, H, phenyl, benzyl, substituted phenyl substituted by halogen, alkyl, alkoxy, hydroxyl or the like, substituted benzyl substituted by halogen, alkyl, alkoxy, hydroxyl or the like;

[0009] R4 and R5 are independently selected from H, halogen, alkyl, substituted alkyl, and trifluoromethyl.

[0010] According to the above scheme, in a preferred case, in the compound described by general formula I, R1 is selected from Wherein: E is selected from an alkyl chain consisting of 1-2 carbon atoms, -NH-, -N(CH3)-, -CO-, -SO2-, G is selected from C, N, R is selected from Cl, F, Br, OH, CH3, CF 3， NH2 and other groups are substituted or polysubstituted; X is selected from N, O, when X is O, R2 does not exist; when X is N, R2 is selected from H, methyl, ethyl, trifluoromethyl; R3 is selected from methyl, ethyl, cyclopropyl, H, phenyl, benzyl, substituted benzyl substituted by halogen, alkyl, alkoxy, hydroxyl, etc., substituted phenyl substituted by halogen, alkyl, alkoxy, hydroxyl, etc. R4, R5 are independently selected from H, F.

[0011] According to the above scheme, under more preferred conditions, in the compound described by general formula I, R1 is selected from Wherein R is amino, hydroxyl, halogen, H, alkyl; X is selected from N, O, when X is O, R2 is absent; when X is N, R2 is selected from H, methyl, ethyl; R3 is selected from methyl, ethyl, cyclopropyl, H, phenyl, benzyl, Substituted benzyl substituted by halogen, alkyl, alkoxy, hydroxyl or the like; substituted phenyl substituted by halogen, alkyl, alkoxy, hydroxyl or the like; R4 is H and R5 is F.

[0012] According to the above scheme, in a particularly preferred case, the compound described by general formula I is selected from at least one of the target compounds in Table 2.

[0013] Table 2

[0014]

[0015]

[0016]

[0017]

[0018] In the present invention, the halogen includes fluorine, chlorine, bromine and iodine.

[0019] In the present invention, the alkyl group includes a straight-chain alkyl group having 1 to 18 carbon atoms and a branched-chain alkyl group having 3 to 18 carbon atoms.

[0020] In the present invention, the alkoxy group includes a straight chain alkoxy group having 1 to 18 carbon atoms and a branched chain alkoxy group having 3 to 18 carbon atoms.

[0021] In the present invention, the substituted alkyl group refers to an alkyl group in which the hydrogen on the carbon atom can be replaced by a halogen.

[0022] The present invention also provides a method for preparing the compound of the above general formula I. If X is N and R2 is not H, the reaction synthesis route is:

[0023]

[0024] Specifically, raw materials Under the action of triethylamine, toluene is used as solvent to react with N, N-dimethylaminoacrylate at 100-120°C for 2-5h to obtain intermediate product I; then it is neutralized with acetic acid and reacted with Amine exchange is performed to obtain intermediate product II, which is then reacted at 100-120°C for a period of time, such as 8-12 hours, under the action of potassium carbonate, and then post-treated (concentrated and washed with water and isopropanol and dried respectively) to obtain intermediate product Ш; intermediate product Ш is reacted at 100-120°C under the action of boric acid, acetic anhydride and zinc chloride, and washed with water and dried to obtain intermediate product IV; intermediate product IV is reacted with dimethyl sulfoxide in an organic solvent The intermediate product V is obtained by reacting at 100-120°C, and then hydrolyzing under alkaline conditions, specifically, reacting under the action of 6% sodium hydroxide solution, adjusting the pH to neutral with glacial acetic acid, washing with water and drying to obtain the intermediate product VI; using dichloromethane as solvent and N,N-dimethylformamide as catalyst, the intermediate product VI is refluxed with thionyl chloride to obtain the intermediate product VII; finally, under the action of triethylamine, reacting with The reaction is carried out in a solvent of dichloromethane for a period of time, such as 3-5 hours, to obtain the final product.

[0025] When X is N and R2 is H, the reaction synthesis route is:

[0026]

[0027] Consistent with the previous steps, the intermediate product The intermediate product IV reacts with piperazine at 100-120° C. in a solvent such as dimethyl sulfoxide to obtain an intermediate product V, and then undergoes a hydrolysis reaction under alkaline conditions. Specifically, the intermediate product IV can be reacted under the action of a 6% sodium hydroxide solution, and the pH is adjusted to neutral with glacial acetic acid, and then washed with water and dried to obtain an intermediate product VI; the intermediate product VI reacts with 9-fluorenylmethyl chloroformate in a solvent of equal proportions of tetrahydrofuran and water under the action of sodium bicarbonate at room temperature for a period of time such as 8-10 hours, and then washed with water and dried to obtain an intermediate product VII; using dichloromethane as a solvent and N,N-dimethylformamide as a catalyst, the intermediate product VII is refluxed with thionyl chloride to obtain an intermediate product VIII; and then, under the action of triethylamine, the intermediate product VII is reacted with thionyl chloride to obtain an intermediate product VIII. The reaction is carried out in a solvent of dichloromethane for a period of time, such as 3-5 hours, and the intermediate product IX is obtained by purification by column chromatography; finally, dichloromethane is used as a solvent and the reaction is carried out under the action of 4-methylpiperidine to obtain the final product.

[0028] When X is O, the reaction synthesis route is:

[0029]

[0030] The same steps as the above reaction are used to obtain intermediate product III, intermediate product III is deesterified in an alcoholic alkaline solution to obtain intermediate product IV, intermediate product IV is reacted with morpholine at 130-150°C in an organic solvent such as dimethyl sulfoxide to obtain intermediate product V; using dichloromethane as solvent and N,N-dimethylformamide as catalyst, intermediate product V is refluxed with thionyl chloride to obtain intermediate product VI; finally, under the action of triethylamine, The reaction is carried out in a solvent of dichloromethane for a period of time, such as 3-5 hours, to obtain the final product.

[0031] It is particularly noted that in some cases, commercial intermediates are available. In the implementation process, the synthesis can be started from the commercial intermediates.

[0032] The present invention also provides the use of the quinolone compounds described in general formula I in the preparation of drugs against DNA viruses such as PRV and / or ASFV.

[0033] According to the above scheme, the above specific application method is: the quinolone compound described in the above general formula I is used as an active ingredient against DNA viruses such as ASFV and PRV, and the content of the active ingredient is 0.1-100% based on the total weight of the antiviral drug.

[0034] According to the above scheme, the specific application method is: use the quinolone compound described in the above general formula I as an active ingredient against ASFV, PRV and other DNA viruses, and add veterinary acceptable carriers and surfactants as needed to prepare hydrates, powders, emulsions, suspensions and granules.

[0035] The third aspect of the present invention provides an anti-PRV and / or ASFV DNA virus drug, comprising an anti-PRV and / or ASFV effective amount of a quinolone compound active ingredient and a pharmaceutically acceptable excipient.

[0036] According to the above scheme, the pharmaceutically acceptable excipient is selected from one or more of solvents, dispersants, diluents, fillers, wetting agents, binders, disintegrants, lubricants, preservatives, suspending agents, emulsifiers, excipients, flavoring agents and carriers.

[0037] According to the above scheme, the dosage form of the drug includes but is not limited to tablets, capsules, granules, pills, liquid preparations, decoctions, suppositories, gels, aerosols or patches.

[0038] Beneficial effects of the present invention:

[0039] The present invention provides a novel quinolone compound represented by general formula I. The quinolone compound represented by general formula I has good inhibition on DNA of PRV, ASFV, etc., can be used for preparing anti-ASFV and / or PRV virus drugs, and has excellent anti-ASFV and PRV activity. DETAILED DESCRIPTION

[0040] Example 1

[0041] C1 (1-cyclopropyl-N-(3,4-dichlorobenzyl)-6-fluoro-4-oxo-7-(piperazin-1-yl)-1,4-dihydroquinoline-3-carboxamide):

[0042]

[0043] NaHCO3 (0.42 g, 5 mmol) was added to a mixture of ciprofloxacin (1.657 g, 5 mmol) in THF-H2O (1: 1). The mixture was stirred for 30 minutes. The reaction mixture was cooled to 0-5 ° C, and 9-fluorenylmethyl chloroformate (1.29 g, 5 mmol) was added to the mixture. The mixture was stirred overnight at ambient temperature. The progress of the reaction was monitored by TLC. The solid was filtered, the mixture was washed with water and dried to obtain the product 7-(4-(((9H-fluorene-9-yl)methoxy)carbonyl)piperazine-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid.

[0044] Dissolve the product 7-(4-(((9H-fluorene-9-yl)methoxy)carbonyl)piperazine-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid (2.7 g, 5 mmol) obtained in the first step in dichloromethane, cool to 0°C, add 1-2 drops of N,N-dimethylformamide, and react for 5 minutes. Add thionyl chloride (1.48 g, 12.5 mmol), heat to 70°C, reflux and stir for 3 hours. After the residual thionyl chloride is removed by rotary evaporation, add dichloromethane to dissolve, cool to 0°C, add triethylamine (0.6 g, 6 mmol) and 3,4-dichlorobenzylamine (1 g, 6 mmol), and react for 3-5 hours. Column chromatography was performed with V(PE):V(EA)=7:3 to obtain the compound (9H-fluoren-9-yl)methyl 4-(1-cyclopropyl-3-((3,4-dichlorobenzyl)carbamoyl)-6-fluoro-4-oxo-1,4-dihydroquinolin-7-yl)piperazine-1-carboxylate.

[0045] The product of the previous step was dissolved in 20% 4-methylpiperidine / dichloromethane and reacted for 1 hour. The progress of the reaction was monitored by TLC, and column chromatography was performed with V(Me2CO):V(DCM)=1:49 to obtain pure compound C1.

[0046] 1 H NMR (400MHz, DMSO-d6) δ10.37(t,J=6.1Hz,1H),8.63(s,1H),7.82(d,J=13.5Hz,1H),7.68–7.53(m,2H),7.46(d,J=7.5Hz,1H),7.32(dd,J=8.3,2. 1Hz,1H),4.53(d,J=6.1Hz,2H),3.73(tt,J=7.2,4.0Hz,1H),3.21–3.13( m,4H),3.00–2.80(m,4H),1.35–1.24(m,2H),1.10(m,J=5.2,4.7Hz,2H).

[0047] MS m / z(ESI):489.125[M+H] + .

[0048] Example 2

[0049] C2 (1-cyclopropyl-6-fluoro-N-(4-methylbenzyl)-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate and 4-methylbenzylamine with reference to C1.

[0050]

[0051] 1 H NMR (400MHz, DMSO-d6) δ10.26(t,J=5.9Hz,1H),8.64(s,1H),7.80(d,J=13.6Hz,1H),7.45(d,J=7.5Hz,1H),7.21(d,J=8.1Hz,2H),7.14(d,J=8.0 Hz,2H),4.49(d,J=5.8Hz,2H),3.77–3.68(m,1H),3.17(t,J=4.9Hz,4H), 2.95–2.83(m,4H),2.28(s,3H),1.32–1.25(m,2H),1.13–1.05(m,2H).MS m / z(ESI):435.219[M+H] + .

[0052] Example 3

[0053] C3 (N-(4-chlorobenzyl)-1-cyclopropyl-6-fluoro-4-oxo-7-(piperazin-1-yl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate and 4-chlorobenzylamine with reference to C1.

[0054]

[0055] 1 H NMR (400MHz, DMSO-d6) δ10.34(t,J=6.0Hz,1H),8.64(s,1H),7.82(d,J=13.6Hz,1H),7.46(d,J=7.5Hz,1H),7.44–7.30(m,4H),4.53(d ,J=6.0Hz,2H),3.73(tt,J=7.2,4.0Hz,1H),3.19(m,J=4.9Hz,4H),3.00–2.84(m,4H),1.33–1.25(m,2H),1.10(q,J=7.1,5.8Hz,2H).MS m / z(ESI):455.164[M+H] +

[0056] Example 4

[0057] C4 (N-(3-chlorobenzyl)-1-cyclopropyl-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate, and 3-chlorobenzylamine with reference to C1.

[0058]

[0059] 1H NMR (400MHz, DMSO-d6) δ10.34(t,J=5.5Hz,1H),8.64(s,1H),7.82(d,J=13.6Hz,1H),7.46(d,J=7.4Hz,1H),7.40–7.33(m,2H),7.30(d,J=12.0Hz ,2H),4.55(d,J=6.0Hz,2H),3.73(tt,J=7.9,7.4,4.0Hz,1H),3.19(t,J =4.9Hz,4H),2.95–2.84(m,4H),1.33–1.25(m,2H),1.14–1.07(m,2H).MS m / z(ESI):455.164[M+H] +

[0060] Example 5

[0061] C5 (1-cyclopropyl-6-fluoro-4-oxo-7-(1-piperazinyl)-N-(pyridin-2-ylmethyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate and 2-aminomethylpyridine with reference to C1.

[0062]

[0063] 1 H NMR (400MHz, DMSO-d6) δ10.50(t,J=5.7Hz,1H),8.63(s,1H),8.55(ddd,J=4.8,1.9,1.0Hz, 1H),7.83(d,J=13.5Hz,1H),7.77(td,J=7.6,1.8Hz,1H),7.46(d,J=7.5Hz,1H),7.35(dd,J =7.8,1.0Hz,1H),7.28(ddd,J=7.5,4.9,1.2Hz,1H),4.65(d,J=5.6Hz,2H),3.73(tt,J=7.1 ,4.1Hz,1H),3.19–3.16(m,4H),2.95–2.85(m,4H),1.33–1.25(m,2H),1.13–1.07(m,2H).MS m / z(ESI):422.198[M+H] +

[0064] Example 6

[0065] C6 (N-(4-chloro-2-fluorobenzyl)-1-cyclopropyl-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate, and 4-chloro-2-fluorobenzylamine with reference to C1.

[0066]

[0067] 1 H NMR (400MHz, DMSO-d6) δ10.33(t,J=6.0Hz,1H),8.62(s,1H),7.83(d,J=13.4Hz,1H),7.47(d,J=7.4Hz,1H),7.45–7.35(m,2H),7.27(dd,J=8.3 ,2.1Hz,1H),4.56(d,J=6.1Hz,2H),3.74(tt,J=7.2,4.1Hz,1H),3.30–3 .22(m,4H),3.08–2.95(m,4H),1.33–1.25(m,2H),1.15–1.05(m,2H).MS m / z(ESI):473.153[M+H] +

[0068] Example 7

[0069] C7 (1-cyclopropyl-6-fluoro-4-oxo-7-(1-piperazinyl)-N-(4-(trifluoromethyl)benzyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate, and 4-(trifluoromethyl)benzylamine with reference to C1.

[0070]

[0071] 1 H NMR (400MHz, DMSO-d6) δ10.42(t,J=6.1Hz,1H),8.64(s,1H),7.82(d,J=13.6Hz,1H),7.70(d,J=8.1Hz,2H),7.54(d,J=8.0Hz,2H),7.46(d,J= 7.5Hz,1H),4.64(d,J=6.0Hz,2H),3.74(tt,J=7.3,4.0Hz,1H),3.21–3.16(m,4H),2.94–2.88(m,4H),1.33–1.25(m,2H),1.13–1.07(m,2H).MS m / z(ESI):489.190[M+H] +

[0072] Example 8

[0073] C8 (N-(5-chloro-2-fluorobenzyl)-1-cyclopropyl-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate, and 5-chloro-2-fluorobenzylamine with reference to C1.

[0074]

[0075] 1 H NMR (400MHz, DMSO-d6) δ10.36(t,J=6.0Hz,1H),8.62(s,1H),7.84(d,J=13.6Hz,1H),7.46(d,J=7.5Hz,1H),7.41–7.35(m,2H),7.27( dd,J=9.6Hz,1H),4.56(d,J=5.9Hz,2H),3.76(tt,1H),3.23–3.16(m,4H),2.94–2.87(m,4H),1.32–1.24(m,2H),1.15–1.07(m,2H).MS m / z(ESI):473.155[M+H] +

[0076] Example 9

[0077] C9 (N-(2-amino-5-chlorobenzyl)-1-cyclopropyl-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate, and 2-amino-5-chlorobenzylamine with reference to C1.

[0078]

[0079] 1 H NMR (400MHz, DMSO-d6) δ10.25(t,J=6.2Hz,1H),8.65(s,1H),7.82(d,J=13.6Hz,1 H),7.46(d,J=7.4Hz,1H),7.09(d,J=2.6Hz,1H),6.97(dd,J=8.5,2.6Hz,1H),6.63 (d,J=8.5Hz,1H),5.33(s,2H),4.38(d,J=6.2Hz,2H),3.73(tt,J=7.0,3.3Hz,1H), 3.21(t,J=4.9Hz,4H),3.01–2.84(m,4H),1.33–1.25(m,2H),1.14–1.07(m,2H).MS m / z(ESI):470.175[M+H] +

[0080] Example 10

[0081] C10 (N-(3-chloro-2-fluorobenzyl)-1-cyclopropyl-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate, and 3-chloro-2-fluorobenzylamine with reference to C1.

[0082]

[0083] 1 H NMR (400MHz, DMSO-d6) δ10.36(t,J=6.1Hz,1H),8.57(s,1H),7.74(d,J=13.4Hz,1H),7.53–7.27(m,3H),7.18(t,J=7.9Hz,1 H),4.60(d,J=5.9Hz,2H),3.77–3.59(m,1H),3.22–3.09(m,4H),2.97–2.76(m,4H),1.33–1.18(m,2H),1.14–0.98(m,2H).MS m / z(ESI):473.155[M+H] +

[0084] Embodiment 11

[0085] C11 (N'-(3-chloro-4-fluorophenyl)-1-cyclopropyl-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxylic acid hydrazide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate, and 3-chloro-4-fluorophenylhydrazine hydrochloride with reference to C1.

[0086]

[0087] 1 H NMR (400MHz, DMSO-d6) δ11.23(d,J=3.1Hz,1H),8.62(s,1H),8.16(d,J=3.1Hz,1H),7.92( d,J=13.5Hz,1H),7.51(d,J=7.5Hz,1H),7.19(t,J=9.1Hz,1H),6.80(dd,J=6.4,2.7Hz,1H ),6.71(ddd,J=9.0,4.0,2.8Hz,1H),3.76(tt,J=7.2,4.0Hz,1H),3.37–3.24(m,4H),3.11 –3.02(m,4H),1.29(dd,J=7.5,5.5Hz,2H),1.19–1.01(m,2H).MSm / z(ESI): 474.148[M+H] +

[0088] Example 12

[0089] C12 (1-cyclopropyl-N-(2,5-dichlorobenzyl)-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate, and 2,5-dichlorobenzylamine with reference to C1.

[0090]

[0091] 1 H NMR (400MHz, DMSO-d6) δ10.41(t,J=6.0Hz,1H),8.63(s,1H),7.85(d,J=13.6Hz,1H),7.56–7.43(m,2H),7.38(d,J=7.4Hz,2H),4 .59(d,J=6.0Hz,2H),3.74(tt,J=7.3,4.0Hz,1H),3.25–3.17(m,4H),2.99–2.85(m,4H),1.33–1.25(m,2H),1.15–1.08(m,2H).MS m / z(ESI):489.125[M+H] +

[0092] Example 13

[0093] C13 (N-((4-chloropyridin-2-yl)methyl)-1-cyclopropyl-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized with reference to C1 from ciprofloxacin, 9-fluorenylmethyl chloroformate and 4-chloro-2-pyridinemethylamine.

[0094]

[0095] 1 H NMR (400MHz, DMSO-d6) δ10.51(t,J=5.8Hz,1H),8.64(s,1H),8.52(d,J=6.1Hz,1H),7.87(d,J=13.5Hz,1H),7.48(d,J=7.5Hz,1H),7.46–7.3 8(m,2H),4.66(d,J=5.7Hz,2H),3.75(tt,J=7.2,4.0Hz,1H),3.28–3.17(m,4H),3.04–2.86(m,4H),1.33–1.25(m,2H),1.16–1.07(m,2H).MS m / z(ESI):456.159[M+H] +

[0096] Embodiment 14

[0097] C14 (N-(4-chlorophenethyl)-1-cyclopropyl-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized with reference to C1 from ciprofloxacin, 9-fluorenylmethyl chloroformate and 4-chlorophenethylamine.

[0098]

[0099] 1 H NMR (400MHz, DMSO-d6) δ9.93(t,J=5.7Hz,1H),8.62(s,1H),7.82(d,J=13.6Hz,1H),7.46(d,J=7.5Hz,1H),7.37–7.26(m,4H),3.73(tt,J=7 .2,4.0Hz,1H),3.57(q,J=6.8Hz,2H),3.23–3.16(m,4H),2.97–2.89(m,4H),2.84(t,J=7.1Hz,2H),1.33–1.26(m,2H),1.13–1.05(m,2H).MS m / z(ESI):469.180[M+H] +

[0100] Embodiment 15

[0101] C15 (1-cyclopropyl-6-fluoro-N-(4-fluorophenethyl)-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized with ciprofloxacin, 9-fluorenylmethyl chloroformate and 4-fluorophenethylamine according to C1.

[0102]

[0103] 1 H NMR(400MHz,DMSO-d6)δ9.93(t,J=5.7Hz,1H),8.62(s,1H),7.82(d,J=13.6 Hz,1H),7.46(d,J=7.5Hz,1H),7.32–7.26(m,2H),7.17–7.04(m,2H),3.73( tt,J=7.2,4.0Hz,1H),3.56(q,J=6.8Hz,2H),3.21(t,J=5.0Hz,4H),2.98–2 .90(m,4H),2.83(t,J=7.1Hz,2H),1.33–1.26(m,2H),1.13–1.06(m,2H).MS m / z(ESI):453.209[M+H] +

[0104] Example 16

[0105] C16 (1-cyclopropyl-6-fluoro-N-(4-methylphenethyl)-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized with ciprofloxacin, 9-fluorenylmethyl chloroformate and 4-methylphenethylamine according to C1.

[0106]

[0107] 1 H NMR(400MHz,DMSO-d6)δ9.97(t,J=5.7Hz,1H),8.67(s,1H),7.86(d,J=13.6 Hz,1H),7.50(d,J=7.5Hz,1H),7.17(q,J=8.0Hz,4H),3.77(tt,J=7.2,4.0Hz ,1H),3.59(q,J=6.8Hz,2H),3.25(dd,J=6.3,3.5Hz,4H),3.01–2.93(m,4H), 2.84(t,J=7.2Hz,2H),2.32(s,3H),1.38–1.31(m,2H),1.18–1.11(m,2H).MS m / z(ESI):449.236[M+H] +

[0108] Embodiment 17

[0109] C17 (N-(3-chlorophenethyl)-1-cyclopropyl-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized with reference to C1 from ciprofloxacin, 9-fluorenylmethyl chloroformate and 3-chlorophenethylamine.

[0110]

[0111] 1 H NMR (400MHz, DMSO-d6) δ9.94(t,J=5.8Hz,1H),8.62(s,1H),7.83(d,J=13.6Hz,1 H),7.47(d,J=7.5Hz,1H),7.34(t,J=2.0Hz,1H),7.32(d,J=7.6Hz,1H),7.29–7.2 1(m,2H),3.74(tt,J=7.2,4.0Hz,1H),3.58(q,J=6.8Hz,2H),3.24–3.19(m,4H),2 .98–2.91(m,4H),2.86(t,J=7.1Hz,2H),1.34–1.26(m,2H),1.13–1.05(m,2H).MS m / z(ESI):469.179[M+H] +

[0112] Embodiment 18

[0113] C18 (N-(3-bromophenethyl)-1-cyclopropyl-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate, and 3-fluorophenethylamine with reference to C1.

[0114]

[0115] 1 H NMR(400MHz,DMSO-d6)δ9.95(t,J=5.8Hz,1H),8.60(s,1H),7.77(d,1H),7.4 3(d,J=7.4Hz,1H),7.38–7.28(m,1H),7.16–7.08(m,2H),7.08–6.97(m,1H), 3.72(tt,J=7.2,4.0Hz,1H),3.57(q,J=6.7Hz,2H),3.22–3.13(m,4H),2.96– 2.88(m,4H),2.86(t,J=7.1Hz,2H),1.33–1.24(m,2H),1.13–1.04(m,2H).MS m / z(ESI):453.211[M+H] +

[0116] Embodiment 19

[0117] C19 (N-(3-bromophenethyl)-1-cyclopropyl-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate, and 3-bromophenethylamine with reference to C1.

[0118]

[0119] 1 H NMR (400MHz, DMSO-d6) δ9.93(t,J=5.8Hz,1H),8.58(s,1H),7.77(d,J=13.6Hz,1H),7.47(s,1H),7.44–7.35(m,2H),7.29–7.21(m,2H),3.70(tt ,J=7.3,4.1Hz,1H),3.56(q,J=6.8Hz,2H),3.23–3.15(m,4H),2.96–2.8 9(m,4H),2.84(t,J=7.2Hz,2H),1.32–1.25(m,2H),1.12–1.03(m,2H).MS m / z(ESI):513.129[M+H]+

[0120] Embodiment 20

[0121] C20 (1-cyclopropyl-6-fluoro-N-(2-fluorophenethyl)-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate, and 2-fluorophenethylamine with reference to C1.

[0122]

[0123] 1 H NMR(400MHz,DMSO-d6)δ9.95(t,J=5.8Hz,1H),8.61(s,1H),7.81(d,J=13.6Hz, 1H),7.45(d,J=7.4Hz,1H),7.38–7.31(m,1H),7.31–7.22(m,1H),7.19–7.09(m ,2H),3.73(tt,J=7.3,4.0Hz,1H),3.57(q,J=6.8Hz,2H),3.25–3.16(m,4H),3. 01–2.91(m,4H),2.88(t,J=7.1Hz,2H),1.33–1.25(m,2H),1.14–1.04(m,2H).MS m / z(ESI):453.208[M+H] +

[0124] Embodiment 21

[0125] C21 (1-cyclopropyl-N-(3,5-dichlorobenzyl)-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from ciprofloxacin, 9-fluorenylmethyl chloroformate, and 3,5-dichlorobenzylamine with reference to C1.

[0126]

[0127] 1 H NMR (400MHz, DMSO-d6) δ10.42(t,J=6.2Hz,1H),8.69(s,1H),7.88(d,J=13.6Hz,1H),7.51(t,J=3.4Hz,2H),7.42(d,J=1.9Hz,2H), 4.60(d,J=6.1Hz,2H),3.78(tt,J=7.4,4.1Hz,1H),3.30–3.18(m,4H),3.04–2.92(m,4H),1.38–1.30(m,2H),1.20–1.12(m,2H).MS m / z(ESI):489.124[M+H]+

[0128] Embodiment 22

[0129] C22 (1-cyclopropyl-N-(3,5-difluorobenzyl)-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized with reference to C1 from ciprofloxacin, 9-fluorenylmethyl chloroformate and 3,5-difluorobenzylamine.

[0130]

[0131] 1 H NMR (400MHz, DMSO-d6) δ10.38(t,J=6.2Hz,1H),8.64(s,1H),7.85(d,J=13.6Hz,1H),7.48(d,J=7.5Hz,1H),7.11(tt,J=9.4,2.4Hz,1H),7.06– 6.99(m,2H),4.56(d,J=6.1Hz,2H),3.74(tt,J=7.1,4.1Hz,1H),3.25–3 .15(m,4H),3.07–2.78(m,4H),1.33–1.25(m,2H),1.16–1.07(m,2H).MS m / z(ESI):457.183[M+H] +

[0132] Embodiment 23

[0133] C23 (1-cyclopropyl-N-(3,4-dichlorobenzyl)-6-fluoro-7-morpholine-4-oxo-1,4-dihydroquinoline-3-carboxamide):

[0134]

[0135] Dissolve cyclopropanecarboxylic acid (1.4 g, 5 mmol) in dimethyl sulfoxide and add morpholine (2.17 g, 25 mmol). Heat to 140°C and react for 1 hour. Cool the solution and concentrate it. -4 The residue was diluted with 1 M HCl and the mixture was extracted with chloroform (3 × 30 ml). -4 The combined organic extracts were washed with 4 M HCl and brine, the filtered mixture was dried and concentrated, and the product, 1-cyclopropyl-6-fluoro-7-morpholinyl-4-oxo-1,4-dihydroquinoline-3-carbonyl chloride, was purified by column chromatography.

[0136] The product of the previous step (1.6 g, 5 mmol) was dissolved in dichloromethane, cooled to 0°C, 1-2 drops of N,N-dimethylformamide were added, and the reaction was continued for 5 minutes. Sulfonyl chloride (1.48 g, 12.5 mmol) was added, and the temperature was raised to 70°C and refluxed with stirring for 3 hours. The residual thionyl chloride was removed by rotary evaporation, and dichloromethane was added to dissolve, and triethylamine (0.6 g, 6 mmol) and 3,4-dichlorobenzylamine (1 g, 6 mmol) were added to cool to 0°C, and the reaction was continued for 3-5 hours. Compound C23 was obtained by column chromatography with V(PE):V(EA)=7:3.

[0137] 1H NMR (400MHz, Chloroform-d) δ10.48(t,J=6.1Hz,1H),8.84(s,1H),8.03(d,J=13.3Hz,1H),7.45(d,J=2.1Hz,1H),7.35(dd,J=18.4,7.6Hz,2H),7.21(d d,J=8.3,2.1Hz,1H),4.60(d,J=6.0Hz,2H),3.98–3.88(m,4H),3.49(tt,J= 7.2,4.0Hz,1H),3.32–3.24(m,4H),1.39–1.31(m,2H),1.22–1.15(m,2H).MS m / z(ESI):490.108[M+H] +

[0138] Embodiment 24

[0139] C24 (N-(3-chlorobenzyl)-1-cyclopropyl-6-fluoro-7-morpholine-4-oxo-1,4-dihydroquinoline-3-carboxamide): synthesized from cyclopropylcarboxylic acid, morpholine and 3-chlorobenzylamine with reference to C23.

[0140]

[0141] 1 H NMR (400MHz, Chloroform-d) δ10.45 (t, J=6.0Hz, 1H), 8.85 (s, 1H), 8.03 (d, J=

[0142] 13.3Hz,1H),7.38–7.29(m,2H),7.29–7.16(m,3H),4.63(d,J=6.0Hz,2H),3.97–3.89(m,4H),3.54 –3.43(m,1H),3.33–3.20(m,4H),1.47–1.28(m,2H),1.22–1.13(m,2H).MSm / z(ESI): 456.147[M+H] +

[0143] Embodiment 25

[0144] C25 (N-(5-chloro-2-fluorobenzyl)-1-cyclopropyl-6-fluoro-7-morpholine-4-oxo-1,4-dihydroquinoline-3-carboxamide): synthesized from cyclopropylcarboxylic acid, morpholine and 5-chloro-2-fluorobenzylamine with reference to C23.

[0145]

[0146] 1 H NMR (400MHz, DMSO-d6) δ10.33(t,J=6.0Hz,1H),8.64(s,1H),7.86(d,J=13.6Hz,1H),7.50(d,J=7.5Hz,1H),7.38(dd,J=7.5,5.0Hz,2H),7.31– 7.21(m,1H),4.57(d,J=6.0Hz,2H),3.86–3.77(m,4H),3.73(tt,J=7.2, 4.0Hz,1H),3.30–3.22(m,4H),1.34–1.26(m,2H),1.19–1.06(m,2H).MS m / z(ESI):474.137[M+H] +

[0147] Embodiment 26

[0148] C26 (N-((4-chloropyridin-2-yl)methyl)-1-cyclopropyl-6-fluoro-7-morpholine-4-oxo-1,4-dihydroquinoline-3-carboxamide): synthesized from cyclopropylcarboxylic acid, morpholine and 4-chloro-2-pyridinemethylamine with reference to C23.

[0149]

[0150] 1 H NMR (400MHz, Chloroform-d) δ10.68(t,J=5.8Hz,1H),8.85(s,1H),8.49(d,J=5.3Hz,1H),8.07(d,J=13.3Hz,1H),7.39–7.30(m,2H),7.18(dd ,J=5.3,2.0Hz,1H),4.80(d,J=5.8Hz,2H),3.97–3.89(m,4H),3.55–3.44(m,1H),3.37–3.25(m,4H),1.45–1.25(m,2H),1.29–1.14(m,2H).MS m / z(ESI):457.142[M+H] +

[0151] Embodiment 27

[0152] C27 (N-(3-chlorobenzyl)-1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxamide): synthesized from cyclopropylcarboxylic acid, N-ethylpiperazine, and 3-chlorobenzylamine with reference to C23. Special note: X is N and R2 is ethyl, so no group protection is required, so the synthesis method is consistent with C23.

[0153]

[0154] 1 H NMR (400MHz, DMSO-d6) δ10.36(t,J=6.2Hz,1H),8.64(s,1H),7.83(d,J=13.5H z,1H),7.48(d,J=7.5Hz,1H),7.40–7.33(m,2H),7.33–7.27(m,2H),4.55(d,J =6.0Hz,2H),3.76–3.69(m,1H),3.30–3.24(m,4H),2.62–2.54(m,4H),2.41(q ,J=7.2Hz,2H),1.33–1.25(m,2H),1.14–1.08(m,2H),1.05(t,J=7.2Hz,3H).MS m / z(ESI):483.192[M+H] +

[0155] Embodiment 28

[0156] C28 (N-(3,4-dichlorobenzyl)-6-fluoro-4-oxo-1-phenyl-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide):

[0157] Dissolve N, N-dimethylaminoacrylate (0.71g, 5mmol) in toluene, then add triethylamine (0.5g, 5mmol) and stir to mix evenly; Take 2,4-dichloro-5-fluorobenzoyl chloride (1.1g, 5mmol), dissolve it in 3ml toluene, then slowly drip the dissolved substance into the reaction bottle, after the dripping is completed, increase the temperature to 110℃, and stir to react for 2-5 hours. After cooling to room temperature, add acetic acid (0.36g, 6mmol) for acidification, drip aniline (0.46g, 5mmol) and continue stirring at room temperature overnight. Monitor the progress of the reaction by TLC, add pure water to transfer the mixture to a separatory funnel, separate the water layer, wash once with distilled water, and extract with toluene. Combine the organic layers into the reaction bottle, add potassium carbonate (0.7g, 5mmol), adjust the temperature to 110℃ and react for 8-12 hours. Toluene was evaporated under reduced pressure to precipitate a brown solid, which was washed three times with water and isopropanol respectively and dried to obtain a solid product, ethyl 7-chloro-6-fluoro-4-oxo-1-phenyl-1,4-dihydroquinoline-3-carboxylate.

[0158] Add boric acid (0.46 g, 7.5 mmol), 3 ml of acetic anhydride and a catalytic amount of zinc chloride (7 mg, 0.05 mmol) to a flask, stir and heat to dissolve, slowly heat to 110 ° C, and keep warm for 30 minutes. Add the product of the previous step (1.73 g, 5 mmol) and keep warm for 2 hours. Monitor the progress of the reaction by TLC, cool, add water for suction filtration, wash three times with distilled water and dry to obtain a solid product 8-chloro-9-fluoro-4-oxo-6-phenyl-4,6-dihydro-2H-1l3,2l4-[1,3,2]dioxaboryl[5,6-c]quinoline-2,2-diyl diacetate.

[0159] The solid product (2.2 g, 5 mmol) from the previous step was dissolved in dimethyl sulfoxide, piperazine (2.1 g, 25 mmol) was added, and refluxed at 110°C for 2 hours. 15 ml of 6% sodium hydroxide solution was added, and the reflux reaction was continued for 2 hours. The reaction solution was adjusted to pH 7 with glacial acetic acid, and refrigerated overnight to precipitate. After suction filtration, the filter cake was placed in a vacuum dryer to dry to obtain a solid product 6-fluoro-4-oxo-1-phenyl-7-(piperazine-1-yl)-1,4-dihydroquinoline-3-carboxylic acid.

[0160] NaHCO3 (0.42 g, 5 mmol) was added to a mixture of the solid product tris (1.83 g, 5 mmol) in THF-H2O (1: 1) from the previous step. The mixture was stirred for 30 minutes. The reaction mixture was cooled to 0-5 ° C, and 9-fluorenylmethyl chloroformate (1.29 g, 5 mmol) was added to the mixture. The mixture was stirred overnight at ambient temperature. The progress of the reaction was monitored by TLC. The solid was filtered, the mixture was washed with water and dried to obtain a solid product 7-(4-(((9H-fluorene-9-yl)methoxy)carbonyl)piperazine-1-yl)-6-fluoro-4-oxo-1-phenyl-1,4-dihydroquinoline-3-carboxylic acid.

[0161] The obtained solid product 7-(4-(tert-butyloxycarbonyl)piperazine-1-yl)-6-fluoro-4-oxo-1-phenyl-1,4-dihydroquinoline-3-carboxylic acid (2.95 g, 5 mmol) was dissolved in dichloromethane, cooled to 0°C, 1-2 drops of N,N-dimethylformamide were added, and the reaction was continued for 5 minutes. Sulfonyl chloride (1.48 g, 12.5 mmol) was added, and the temperature was raised to 70°C and refluxed for 3 hours. After the residual thionyl chloride was removed by rotary evaporation, dichloromethane was added to dissolve, and triethylamine (0.6 g, 6 mmol) and 3,4-dichlorobenzylamine (1 g, 6 mmol) were added to cool to 0°C, and the reaction was continued for 3-5 hours. Column chromatography was performed with V(PE):V(EA)=7:3 to obtain (9H-fluoren-9-yl)methyl 4-(3-((3,4-dichlorobenzyl)carbamoyl)-6-fluoro-4-oxo-1-phenyl-1,4-dihydroquinolin-7-yl)piperazine-1-carboxylate.

[0162] The product was dissolved in 20% 4-methylpiperidine / dichloromethane and reacted for 1 hour. The progress of the reaction was monitored by TLC, and column chromatography was performed with V(Me2CO):V(DCM)=1:49 to obtain C28.

[0163] 1 H NMR (400MHz, Chloroform-d) δ10.48(t,J=6.1Hz,1H),8.84(s,1H),8.03(d,J=13.3Hz,1H),7.45(d,J=2.1Hz,1H),7.35(dd,J=18.4,7.6Hz,2H),7.21(d d,J=8.3,2.1Hz,1H),4.60(d,J=6.0Hz,2H),3.98–3.88(m,4H),3.49(tt,J= 7.2,4.0Hz,1H),3.32–3.24(m,4H),1.39–1.31(m,2H),1.22–1.15(m,2H).MS m / z(ESI):525.123[M+H] +

[0164] Embodiment 29

[0165] C29 (N-(3-chlorobenzyl)-6-fluoro-4-oxo-1-phenyl-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from N,N-dimethylaminoacrylate, 2,4-dichloro-5-fluorobenzoyl chloride, aniline, piperazine, 9-fluorenylmethyl chloroformate, and 3-chlorobenzylamine with reference to C28.

[0166]

[0167] 1 H NMR (400MHz, DMSO-d6) δ10.37(t,J=6.1Hz,1H),8.52(s,1H),7.91(d,J=13.5Hz,1H),7.73–7.66(m,5H), 7.40–7.29(m,4H),6.36(d,J=7.4Hz,1H),4.57(d,J=6.1Hz,2H),2.93–2.82(m,4H),2.81–2.71(m,4H).MS m / z(ESI): 491.162[M+H] +

[0168] Embodiment 30

[0169] C30 (N-(5-chloro-2-fluorobenzyl)-6-fluoro-4-oxo-1-phenyl-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from N,N-dimethylaminoacrylate, 2,4-dichloro-5-fluorobenzoyl chloride, aniline, piperazine, 9-fluorenylmethyl chloroformate, and 5-chloro-2-fluorobenzylamine with reference to C28.

[0170]

[0171] 1 H NMR (400MHz, DMSO-d6) δ10.37(t,J=6.0Hz,1H),8.51(s,1H),7.93(d,J=13.4Hz,1H),7.74–7.61(m,5H),7.45–7.3 4(m,2H),7.34–7.20(m,1H),6.36(d,J=7.3Hz,1H),4.59(d,J=6.0Hz,2H),2.93–2.83(m,4H),2.81–2.70(m,4H).MS m / z(ESI):509.152[M+H] +

[0172] Embodiment 31

[0173] C31 (N-((4-chloropyridin-2-yl)methyl)-6-fluoro-4-oxo-1-phenyl-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from N,N-dimethylaminoacrylate, 2,4-dichloro-5-fluorobenzoyl chloride, aniline, piperazine, 9-fluorenylmethyl chloroformate, and 4-chloro-2-pyridinemethylamine with reference to C28.

[0174]

[0175] 1 H NMR (400MHz, DMSO-d6) δ10.53(t,J=5.8Hz,1H),8.59–8.46(m,2H),7.93(d,J=13.4Hz,1H),7.76–7.58(m,5H ),7.56–7.38(m,2H),6.36(d,J=7.3Hz,1H),4.68(d,J=5.7Hz,2H),2.92–2.85(m,4H),2.81–2.75(m,4H).MS m / z(ESI): m / z=492.157[M+H] +

[0176] Embodiment 32

[0177] C32 (1-benzyl-N-(5-chloro-2-fluorobenzyl)-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from N,N-dimethylaminoacrylate, 2,4-dichloro-5-fluorobenzoyl chloride, benzylamine, piperazine, 9-fluorenylmethyl chloroformate, and 5-chloro-2-fluorobenzylamine with reference to C28.

[0178]

[0179] 1 H NMR (400MHz, DMSO-d6) δ10.42(t,J=6.1Hz,1H),9.04(s,1H),7.85(d,J=13.5Hz,1H),7.45–7.35(m,4H),7.33–7 .25(m,4H),6.98(d,J=7.2Hz,1H),5.76(s,2H),4.59(d,J=6.1Hz,2H),3.07–2.95(m,4H),2.87–2.77(m,4H).MS m / z(ESI):523.16822[M+H] +

[0180] Embodiment 33

[0181] C33 (N-(3-chlorobenzyl)-1-ethyl-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from N,N-dimethylaminoacrylate, 2,4-dichloro-5-fluorobenzoyl chloride, ethylamine, piperazine, 9-fluorenylmethyl chloroformate, and 3-chlorobenzylamine with reference to C28.

[0182]

[0183] 1 H NMR (400MHz, DMSO-d6) δ10.42(t,J=6.2Hz,1H),8.80(s,1H),7.87(d,J=13.6Hz,1H),7.42–7.27(m,4H),7.08(d,J=7. 3Hz,1H),4.56(d,J=6.0Hz,2H),4.50(q,J=7.2Hz,2H),3.25–3.16(m,4H),2.96–2.87(m,4H),1.40(t,J=7.1Hz,3H).MS m / z(ESI):443.162[M+H] +

[0184] Embodiment 34

[0185] C34 (N'-(3-chlorophenyl)-1-ethyl-6-fluoro-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carbohydrazide): synthesized from N,N-dimethylaminoacrylate, 2,4-dichloro-5-fluorobenzoyl chloride, ethylamine, piperazine, 9-fluorenylmethyl chloroformate, and 3-chlorophenylhydrazine hydrochloride with reference to C28.

[0186]

[0187] 1 H NMR (400MHz, DMSO-d6) δ11.38(d,J=2.8Hz,1H),8.85(s,1H),8.30(d,J=2.8Hz,1H),7.98(d,J=13.7Hz,1H),7.22(t,J=7.8Hz,1H) ,7.15(d,J=7.3Hz,1H),6.83–6.71(m,3H),4.58(q,J=7.1Hz,2H),3.30–3.21(m,4H),2.99–2.91(m,4H),1.47(t,J=7.1Hz,3H).MS m / z(ESI):444.157[M+H] +

[0188] Embodiment 35

[0189] C35 (1-ethyl-6-fluoro-N-(4-hydroxyphenethyl)-4-oxo-7-(1-piperazinyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from N,N-dimethylaminoacrylate, 2,4-dichloro-5-fluorobenzoyl chloride, ethylamine, piperazine, 9-fluorenylmethyl chloroformate, and 4-hydroxyphenethylamine with reference to C28.

[0190]

[0191] 1 H NMR (400MHz, DMSO-d6) δ9.97(t,J=5.7Hz,1H),9.13(s,1H),8.75(s,1H),7.85(d,J=13.6Hz,1H),7.14–6.91(m,3H),6.78–6.55(m,2H ),4.48(q,J=7.1Hz,2H),3.50(q,J=6.9Hz,2H),3.23–3.13(m,4H),2.97–2.84(m,4H),2.71(t,J=7.2Hz,2H),1.38(t,J=7.1Hz,3H).MS m / z(ESI):439.212[M+H] +

[0192] Embodiment 36

[0193] C36 (N-(5-chloro-2-fluorobenzyl)-6-fluoro-1-(4-fluorophenyl)-4-oxo-7-(piperazin-1-yl)-1,4-dihydroquinoline-3-carboxamide): synthesized from N,N-dimethylaminoacrylate, 2,4-dichloro-5-fluorobenzoyl chloride, 4-fluoroaniline, piperazine, 9-fluorenylmethyl chloroformate, and 5-chloro-2-fluorobenzylamine with reference to C28.

[0194]

[0195] 1 H NMR (400MHz, DMSO-d6) δ10.36(t,J=6.1Hz,1H),8.50(s,1H),7.91(d,J=13.4Hz,1H),7.83–7.71(m,2H),7.60–7.48(m,2H),7.43–7. 35(m,2H),7.35–7.22(m,1H),6.32(d,J=7.3Hz,1H),4.59(d,J=6.0Hz,2H),2.92(m,J=6.5,3.4Hz,4H),2.81(m,J=6.4,3.5Hz,4H).MS m / z(ESI): m / z=527.148[M+H] +

[0196] Embodiment 37

[0197] C37 (N-(5-chloro-2-fluorobenzyl)-6-fluoro-1-(2-fluorophenyl)-4-oxo-7-(piperazin-1-yl)-1,4-dihydroquinoline-3-carboxamide): synthesized from N,N-dimethylaminoacrylate, 2,4-dichloro-5-fluorobenzoyl chloride, o-fluoroaniline, piperazine, 9-fluorenylmethyl chloroformate, and 5-chloro-2-fluorobenzylamine with reference to C28.

[0198]

[0199] 1 H NMR (400MHz, DMSO-d6) δ10.29(t,J=6.1Hz,1H),8.58(s,1H),7.93(d,J=13.4H z,1H),7.84(td,J=7.8,1.9Hz,1H),7.82–7.72(m,1H),7.64(ddd,J=9.9,8.3,1 .4Hz,1H),7.53(td,J=7.7,1.4Hz,1H),7.46–7.35(m,2H),7.33–7.21(m,1H), 6.30–6.22(m,1H),4.65–4.52(m,2H),2.97–2.86(m,4H),2.83–2.70(m,4H).MS m / z(ESI): m / z=527.145[M+H] +

[0200] Embodiment 38

[0201] C38 (N-(5-chloro-2-fluorobenzyl)-6-fluoro-4-oxo-7-(piperazin-1-yl)-1-(p-tolyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from N,N-dimethylaminoacrylate, 2,4-dichloro-5-fluorobenzoyl chloride, p-toluidine, piperazine, 9-fluorenylmethyl chloroformate, and 5-chloro-2-fluorobenzylamine with reference to C28.

[0202]

[0203] 1H NMR (400MHz, DMSO-d6) δ10.37(t,J=6.1Hz,1H),8.47(s,1H),7.91(d,J=13.4Hz,1H),7.56(d,J=8.3Hz,2H),7.48(d,J=8.5Hz,2H),7.4 3–7.35(m,2H),7.35–7.22(m,1H),6.39(d,J=7.4Hz,1H),4.58(d,J=6.0Hz,2H),2.95–2.83(m,4H),2.83–2.71(m,4H),2.46(s,3H).MS m / z(ESI): m / z=523.173[M+H] +

[0204] Embodiment 39

[0205] C39 (N-(5-chloro-2-fluorobenzyl)-6-fluoro-1-(4-methoxyphenyl)-4-oxo-7-(piperazin-1-yl)-1,4-dihydroquinoline-3-carboxamide): synthesized from N,N-dimethylaminoacrylate, 2,4-dichloro-5-fluorobenzoyl chloride, p-aminoanisole, piperazine, 9-fluorenylmethyl chloroformate, and 5-chloro-2-fluorobenzylamine with reference to C28.

[0206]

[0207] 1 H NMR (400MHz, DMSO-d6) δ10.37(t,J=6.1Hz,1H),8.47(s,1H),7.91(d,J=13.4Hz,1H),7.65–7.56(m,2H),7.39(dd,J=7.3,4.9Hz,2H),7. 32–7.24(m,1H),7.24–7.15(m,2H),6.39(d,J=7.4Hz,1H),4.58(d,J=6.0Hz,2H),3.88(s,3H),2.96–2.86(m,4H),2.86–2.76(m,4H).MS m / z(ESI): m / z=539.165[M+H] +

[0208] Embodiment 40

[0209] C40 (N-(5-chloro-2-fluorobenzyl)-6-fluoro-4-oxo-7-(piperazin-1-yl)-1-(pyridin-2-ylmethyl)-1,4-dihydroquinoline-3-carboxamide): synthesized from N,N-dimethylaminoacrylate, 2,4-dichloro-5-fluorobenzoyl chloride, 2-aminomethylpyridine, piperazine, 9-fluorenylmethyl chloroformate, and 5-chloro-2-fluorobenzylamine with reference to C28.

[0210]

[0211] 1 H NMR (400MHz, DMSO-d6) δ10.42(t,J=6.1Hz,1H),9.03(s,1H),8.50(dd,J=4.9,2.2Hz,1H),7.89–7.78(m,2H),7.53(d,J=7.8Hz,1 H),7.47–7.37(m,2H),7.37–7.24(m,2H),7.01(d,J=7.3Hz,1H),4.60(d,J=6.0Hz,2H),3.05–2.93(m,4H),2.87–2.74(m,4H).MS m / z(ESI): m / z=524.166[M+H] +

[0212] Embodiment 41

[0213] Taking compound C1 as an example, the anti-PRV activity experiment is as follows:

[0214] The porcine kidney cell line PK15 cells were maintained in our laboratory and were cultured with fresh Dulbecco's modified eagle medium (DMEM) (No. 11965092, Gibco TM ), 10% fetal bovine serum (FBS) (No.: P30-3306, PAN TM ) and 1% penicillin-streptomycin solution (100X, No. BL505A, Biosharp TM ) were cultured at 37°C.

[0215] PK15 cells with 80% confluence were cultured in 96-well plates. After 24 hours, the culture medium was discarded, and the cells were rinsed 3 times with PBS. 1 mL of fresh DMEM containing 2% FBS and 1% penicillin-streptomycin solution was added and cultured at 37°C for 4 hours. Then, the cells were treated with different concentrations (1 μM, 0.5 μM, 0.25 μM, 0.1 μM, 0.05 μM, 0.01 μM, 0.005 μM, 0.001 μM) of C1 compound for 2 hours. The cells were then infected with PRV (MOI = 1), and total DNA was extracted with phenol-chloroform (Solabio Biotech Co., Ltd.) after 48 hours. The target gene was amplified by SYBR green-based quantitative PCR (qPCR) using 2×Taq Master Mix (Vazyme, Nanjing, China). The Ct value of each well was determined using a real-time fluorescence quantitative PCR instrument.

[0216] Virus control wells: The difference from the drug-treated wells is that no test solution is added.

[0217] The effect of the drug on the virus replication copy number in each test well was calculated using the standard curve: Y=-3.4395X+43.179 (R2=0.999), where x is lg (virus copy number) and y is the Ct value.

[0218] Table 3 lists the primers used in this study. The inhibition rate calculation formula for the experimental group is

[0219]

[0220] EC was obtained by nonlinear regression fitting of the curve using the logistic equation in Origin software. 50 .

[0221] Table 3

[0222]

[0223] Embodiment 42

[0224] Taking compound C1 as an example, the anti-ASFV activity experiment is as follows:

[0225] Vero cells were cultured at 5×10 4 Cells / well (volume 100 μL) were inoculated in a 96-well cell culture plate. When the cells adhered and the confluence rate exceeded 90%, they were replaced with DMEM maintenance medium (DMEM medium, 2% serum, penicillin 100U / mL, streptomycin 100μg / mL). ASFV virus of 1MOI (multiplicity of infection) was added to the infection group. At the same time, different final concentrations (1μM, 0.5μM, 0.25μM, 0.1μM, 0.05μM, 0.01μM, 0.005μM, 0.001μM) of C1 compounds were added and cultured in a 37°C cell culture incubator for 48h. Take out the culture plate and freeze and thaw once at -20°C, blow each well and draw 200μL to add to the virus nucleic acid purification kit machine to automatically extract DNA. The Ct value of each well was determined using a real-time fluorescence quantitative PCR instrument.

[0226] Virus control wells: The difference from the drug-treated wells is that no test solution is added.

[0227] Through the standard curve: y = -3.7897x + 47.997 (R 2 =0.9959), x is lg (virus copy number), y is Ct value, and the effect of drug on virus replication copy number in each test well is calculated.

[0228] Table 4 lists the primers used in this study.

[0229] Virus inhibition rate (%) = (virus copy number in virus control group - virus copy number in drug administration group) / virus copy number in virus control group × 100%

[0230] EC was obtained by nonlinear regression fitting of the curve using the logistic equation in Origin software. 50

[0231] Table 4

[0232]

[0233] Embodiment 43

[0234] Anti-DNA virus activity of compounds

[0235] Table 5 Anti-DNA virus activity of compounds

[0236]

[0237]

[0238] A:<1nM

[0239] B:1-10nM

[0240] C:10-100nM

[0241] D:100-1000nM

[0242] E>1uM

[0243] ND:not determined

[0244] It should be understood that those skilled in the art can make improvements or changes based on the above description, and all these improvements and changes should fall within the scope of protection of the appended claims of the present invention.

Claims

1. The quinolone compound described in general formula I is characterized in that: The general formula I is: Wherein R1 is selected from Wherein Z is selected from an alkyl chain consisting of 1 to 4 carbon atoms, -NH-, -N(CH3)-, -CO-, -SO2-, and Y is selected from Aromatic ring structures and the above aromatic ring structures substituted with -OH, halogen, CH3, CF3, NH2, and alkoxy groups; X is selected from N, O, when X is O, R2 is absent, when X is N, R2 is selected from H, alkyl, substituted alkyl, trifluoromethyl; R3 is selected from alkyl, cycloalkyl, H, phenyl, benzyl, substituted phenyl substituted by halogen, alkyl, alkoxy or hydroxyl groups, substituted benzyl substituted by halogen, alkyl, alkoxy or hydroxyl groups; R4 and R5 are independently selected from H, halogen, alkyl, substituted alkyl, and trifluoromethyl.

2. The quinolone compound according to claim 1, characterized in that: In the compound described in the general formula I, R1 is selected from Wherein: E is selected from an alkyl chain consisting of 1-2 carbon atoms, -NH-, -N(CH3)-, -CO-, -SO2-, G is selected from C, N, R is selected from Cl, F, Br, OH, CH3, CF 3， NH2 group is monosubstituted or polysubstituted; X is selected from N, O, when X is O, R2 is absent; when X is N, R2 is selected from H, methyl, ethyl, trifluoromethyl; R3 is selected from methyl, ethyl, cyclopropyl, H, phenyl, benzyl, substituted benzyl substituted by halogen, alkyl, alkoxy, hydroxyl group, substituted phenyl substituted by halogen, alkyl, alkoxy, hydroxyl group. R4, R5 are independently selected from H, F.

3. The quinolone compound according to claim 1, characterized in that: In the compound described in the general formula I, R1 is selected from Wherein R is amino, hydroxyl, halogen, H, alkyl; X is selected from N, O, when X is O, R2 is absent; when X is N, R2 is selected from H, methyl, ethyl; R3 is selected from methyl, ethyl, cyclopropyl, H, phenyl, benzyl, substituted benzyl substituted by halogen, alkyl, alkoxy or hydroxyl groups; substituted phenyl substituted by halogen, alkyl, alkoxy or hydroxyl groups; R4 is H and R5 is F.

4. The quinolone compound according to claim 1, characterized in that: The compound of formula I is selected from at least one of the target compounds in Table 2: Table 2:

5. The quinolone compound according to claim 1, characterized in that: The halogen includes fluorine, chlorine, bromine and iodine, the alkyl includes a straight-chain alkyl with a carbon number of 1-18 and a branched-chain alkyl with a carbon number of 3-18, the alkoxy includes a straight-chain alkoxy with a carbon number of 1-18 and a branched-chain alkoxy with a carbon number of 3-18, and the substituted alkyl refers to a halogen replacing the hydrogen on the carbon atom in the alkyl.

6. Use of the quinolone compound according to any one of claims 1 to 5 in the preparation of anti-PRV and / or ASFV DNA virus drugs.

7. The use according to claim 6, characterized in that: The specific application method is: using the quinolone compound described in claim 1 as an active ingredient against ASFV and / or PRV DNA virus, and the content of the active ingredient is 0.1-100% based on the total weight of the antiviral drug.

8. The use according to claim 6, characterized in that: The specific application method is: using the quinolone compound described in claim 1 as an active ingredient against ASFV and / or PRV DNA virus, and adding veterinary acceptable carriers and surfactants as needed to prepare a hydrate, powder, emulsion, suspension or granule dosage form.

9. An anti-PRV and / or ASFV DNA virus drug, characterized in that: It comprises an effective amount of the quinolone compound active ingredient according to claim 1 for resisting PRV and / or ASFV and a pharmaceutically acceptable excipient.

10. The anti-PRV and / or ASFV DNA virus drug according to claim 9, characterized in that: The dosage form of the drug includes, but is not limited to, tablets, capsules, granules, pills, liquid preparations, decoctions, suppositories, gels, aerosols or patches.

Images