CN109251211A

CN109251211A - A kind of quinolonecarboxylic acid compound and its intermediate, preparation method and application

Info

Publication number: CN109251211A
Application number: CN201810757136.5A
Authority: CN
Inventors: 刘博�; 徐广宇; 喻杰; 吴玉娥; 鲁洋; 徐方方; 张钰; 韩晓东; 林冬玲; 黄韧; 陈茶; 陆金健; 张玉琴
Original assignee: Guangdong Hospital of Traditional Chinese Medicine; Guangdong Laboratory Animals Monitoring Institute
Current assignee: Guangdong Hospital of Traditional Chinese Medicine; Guangdong Laboratory Animals Monitoring Institute
Priority date: 2016-12-21
Filing date: 2016-12-21
Publication date: 2019-01-22
Also published as: CN106674254A; CN106674254B

Abstract

The invention discloses a kind of quinolonecarboxylic acid compound and its hydrochlorides, also disclose the preparation method of such compound and its hydrochloride, and, such compound and its hydrochloride are preparing the application in resisting gram-positive bacteria or anti-Gram negative bacteria drugs.Also, disclose the preparation method of the intermediate and the intermediate that are used to prepare such compound.

Description

A kind of quinolonecarboxylic acid compound and its intermediate, preparation method and application

Technical field

The present invention relates to quinolone compound, preparation method and its applications, and the invention further relates to for making The intermediate of the standby quinolone compound.

Background technique

The discovery of acidum nalidixicum in 1962 opens a fan gate for the research of quinolone.Quinolone drugs has because of it The advantages that antimicrobial spectrum compare Guang ﹑ activity is strong, is not apparent from crossing drug resistant with other antimicrobials, convenient drug administration, and obtain Pharmaceutical Chemist Blueness narrow.It was found that show very strong neurotoxicity when quinolone side chain is non-substituted piperazine, and by piperazine cycloalkanes After baseization modification, neurotoxicity is substantially reduced (such as application No. is 200310122707.1 Chinese patent applications).

However, it is extensive and unreasonable due to people's medication, cause many bacterium to produce drug resistance to such drug, such as: Gram-positive bacteria Methicillin-resistant Staphylococcus aureus/form staph, penicillin resistant streptococcus etc..Therefore, new the having of design synthesis is lived Property or the lower quinolone carboxylic acid derivatives of side effect it is extremely urgent, to cope with the following need to antibiotics It asks.

Summary of the invention

The purpose of the present invention is to provide a kind of quinolonecarboxylic acid compounds, to solve the above technical problems at least one It is a.

Another object of the present invention, which also resides in, provides the preparation method of above-mentioned quinolonecarboxylic acid compound, above-mentioned to solve At least one of technical problem.

Another object of the present invention, which also resides in, provides the application of above-mentioned quinolonecarboxylic acid compound, to solve above-mentioned technology At least one of problem.

Another object of the present invention, which also resides in, provides a kind of intermediate for being used to prepare above-mentioned quinolonecarboxylic acid compound, At least one of to solve the above technical problems.

Another object of the present invention, which also resides in, provides a kind of intermediate for being used to prepare above-mentioned quinolonecarboxylic acid compound Preparation method, at least one of to solve the above technical problems.

According to an aspect of the present invention, the present invention provides a kind of quinolonecarboxylic acid compound, be structure such as formula (I), (II), one or more of (III), (IV), (V) and (VI) compound represented, or for as formula (I), (II), (III) and one or more of the nontoxic salts that are formed of (IV) compound represented:

Wherein, the R in formula (I)₁It can be 2,4-F₂C₆H₃(2,4- difluorophenyls, structural formula are as follows:) or c-C₃H₅ (cyclopropyl, structural formula are as follows:)；R₂It can be (S/R)-CH₂OH or (S/R)-CHOHCH₃；R₃It can be Boc (tertiary butyloxycarbonyl Base, structural formula are as follows:) or H.

R in formula (II)₄It can be (S/R)-CH₂OH or (S/R)-CHOHCH₃；R₅It can be Boc (tertbutyloxycarbonyl, knot Structure formula are as follows:) or H.

R in formula (III)₆It can be H or OH.

R in formula (IV)₇It can be H or OH.

R in formula (V)₈It can be 2,4-F₂C₆H₃(2,4- difluorophenyls, structural formula are as follows:) or c-C₃H₅(cyclopropyl Base, structural formula are as follows:)；R₉It can be H or OH.

R in formula (VI)₁₀It can be 2,4-F₂C₆H₃(2,4- difluorophenyls, structural formula are as follows:) or c-C₃H₅(cyclopropyl Base, structural formula are as follows:)；R₁₁It can be H or OH.

Above-mentioned quinolonecarboxylic acid compound, can manufactured medically acceptable soluble-salt.Wherein can medically it connect The soluble-salt received include the derivative and inorganic acid such as: hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, or such as with organic acid: second Acid, trifluoroacetic acid, citric acid, maleic acid, oxalic acid, succinic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, ascorbic acid, The salt of the formation such as malic acid, methanesulfonic acid, p-methyl benzenesulfonic acid, lysine, aspartic acid.

According to another aspect of the present invention, the present invention provides the preparation methods of above-mentioned quinolonecarboxylic acid compound.

Work as R₁For 2,4-F₂C₆H₃(2,4- difluorophenyls,), R₂For (S/R)-CH₂OH, R₃For Boc (tertbutyloxycarbonyl,) when, formula (I) compound and its hydrochloride can be made by following steps:

The fluoro- 1,4- dihydro -4- Oxoquinoline-3-carboxylic acid of 1- (2,4 difluorobenzene base) -6,7,8- three is dissolved in DMSO (diformazan Base sulfoxide) in, TEA (triethanolamine) and 1- tertbutyloxycarbonyl -2- methylol piperazine is added, it is micro- under conditions of 80~100 DEG C Wave reacts 10~14h, is cooled to room temperature, and methylene chloride is added, then wash 1~3 time, dry, is recrystallized with dehydrated alcohol to obtain the final product To formula (I) compound；

Formula obtained (I) compound is added to the in the mixed solvent of HCl- ethyl alcohol, wherein the concentration of HCl is 2~8mol/ The volume ratio of L, HCl and ethyl alcohol is 1:3~3:1, and outer temperature is heated to 110~120 DEG C of 10~12h of reflux, is cooled to room temperature, and is analysed White solid out is filtered, then is washed 1~3 time with dehydrated alcohol；Obtained solid is added in dehydrated alcohol, and outer temperature is heated to 90 ~120 DEG C of 30~50min of reflux, are cooled to room temperature, and filter, and filter residue is washed again with dehydrated alcohol to get formula (I) compound Hydrochloride.

Work as R₁For 2,4-F₂C₆H₃(2,4- difluorophenyls,), R₂For (S/R)-CHOHCH₃, R₃For Boc (tertiary butyloxycarbonyl Base,) when, formula (I) compound and its hydrochloride can be made by following steps:

The fluoro- Isosorbide-5-Nitrae of 1- (2,4- difluorophenyl) -6,7,8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid is dissolved in DMSO, is added Enter N- methylmorpholine and 1- tertbutyloxycarbonyl -2- (1- ethoxy) piperazine, the microwave reaction 10 under conditions of 80~100 DEG C ~14h, is cooled to room temperature, and methylene chloride is added, then wash 1~3 time, dry, obtains formula (I) with dehydrated alcohol recrystallization Compound；

Formula obtained (I) compound is added to the in the mixed solvent of HCl- ethyl alcohol, wherein the concentration of HCl is 2~8mol/ The volume ratio of L, HCl and ethyl alcohol is 1:3~3:1, and outer temperature is heated to 110~120 DEG C of 10~12h of reflux, is cooled to room temperature, and is analysed White solid out is filtered, then is washed 1~3 time with dehydrated alcohol；Obtained solid is added in dehydrated alcohol, and outer temperature is heated to 90~ 120 DEG C or more 30~50min of reflux, are cooled to room temperature, and filter, and filter residue is washed again with dehydrated alcohol to get formula (I) compound Hydrochloride.

Work as R₁For c-C₃H₅(cyclopropyl,), R₂For (S/R)-CHOHCH₃, R₃For Boc (tertbutyloxycarbonyl,) When, formula (I) compound and its hydrochloride can be made by following steps:

By 1- cyclopropyl -6,7, the fluoro- Isosorbide-5-Nitrae of 8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid is dissolved in DMSO, be added TEA and 1- tertbutyloxycarbonyl -2- (1- ethoxy) piperazine, 10~14h of microwave reaction under conditions of 80~100 DEG C are cooled to room temperature, Methylene chloride is added, then washes 1~3 time, it is dry, formula (I) compound is obtained with dehydrated alcohol recrystallization；

Work as R₁For c-C₃H₅(cyclopropyl,), R₂For (S/R)-CH₂OH, R₃For Boc (tertbutyloxycarbonyl,) when, Formula (I) compound and its hydrochloride can be made by following steps:

By 1- cyclopropyl -6,7, the fluoro- Isosorbide-5-Nitrae of 8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid is dissolved in DMSO, be added TEA and 1- tertbutyloxycarbonyl -2- methylol piperazine, 10~14h of microwave reaction under conditions of 80~100 DEG C are cooled to room temperature, and are added Methylene chloride, then wash 1~3 time, it is dry, formula (I) compound is obtained with dehydrated alcohol recrystallization；

Work as R₄For (S/R)-CH₂OH, R₅For Boc (tertbutyloxycarbonyl,) when, formula (II) compound and its hydrochloride It can be made by following steps:

Lavo-ofloxacin carboxylic acid is dissolved in DMSO, TEA and 1- tertbutyloxycarbonyl -2- methylol piperazine is added, 80~100 20~26h of microwave reaction under conditions of DEG C is cooled to room temperature after the reaction was completed, methylene chloride is added, then wash 1~3 time, is done It is dry, it is recrystallized with dehydrated alcohol up to formula (II) compound；

Formula (II) compound is added to the in the mixed solvent of HCl- ethyl alcohol, wherein the concentration of HCl is 2~8mol/L, HCl Volume ratio with ethyl alcohol is 1:3~3:1, and outer temperature is heated to 100~120 DEG C of 10~12h of reflux, is cooled to room temperature, and white is precipitated Solid filters, and filter residue washs 1~3 time with dehydrated alcohol again, and obtained solid is added in dehydrated alcohol, and outer temperature is heated to 90~ 120 DEG C or more 30~50min of reflux, are cooled to room temperature, and filter, and filter residue washs 1~3 time with dehydrated alcohol again to get formula (II) The hydrochloride of compound.

Work as R₄For (S/R)-CHOHCH₃, R₅For Boc (tertbutyloxycarbonyl,) when, formula (II) compound and its hydrochloric acid Salt can be made by following steps:

Lavo-ofloxacin carboxylic acid is dissolved in DMSO, TEA and 11- tertbutyloxycarbonyl -2- (1- ethoxy) piperazine is added, 80 20~26h of microwave reaction is cooled to room temperature after the reaction was completed under conditions of~100 DEG C, methylene chloride is added, then wash 1~3 It is secondary, it is dry, it is recrystallized with dehydrated alcohol up to formula (II) compound；

Work as R₆When for H, formula (III) compound can be made by following steps:

Lavo-ofloxacin carboxylic acid is dissolved in DMSO, TEA and (R)-octahydro pyrrolo- [1,2-a] pyrazine is added, 80~100 20~26h of microwave reaction under conditions of DEG C is cooled to room temperature after the reaction was completed, methylene chloride is added, then wash 1~3 time, is done It is dry, it is recrystallized with dehydrated alcohol up to formula (III) compound.

Formula (III) compound is added to the in the mixed solvent of HCl- ethyl alcohol, wherein the concentration of HCl is 2~8mol/L, HCl Volume ratio with ethyl alcohol is 1:3~3:1, and outer temperature is heated to 100~120 DEG C of 10~12h of reflux, is cooled to room temperature, and white is precipitated Solid filters, and filter residue washs 1~3 time with dehydrated alcohol again, and obtained solid is added in dehydrated alcohol, and outer temperature is heated to 90~ 120 DEG C or more 30~50min of reflux, are cooled to room temperature, and filter, and filter residue washs 1~3 time with dehydrated alcohol again to get formula (III) The hydrochloride of compound.

Work as R₆When for OH, formula (III) compound can be made by following steps:

Lavo-ofloxacin carboxylic acid is dissolved in DMSO, and TEA and (S/R) -7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazine is added, 20~26h of microwave reaction is cooled to room temperature after the reaction was completed under conditions of 80~100 DEG C, methylene chloride is added, then wash 1 It is~3 times, dry, it is recrystallized with dehydrated alcohol up to formula (III) compound.

Work as R₇When for H, formula (IV) compound can be made by following steps:

Lavo-ofloxacin carboxylic acid is dissolved in DMSO, TEA and (S/R)-octahydro pyrrolo- [1,2-a] pyrazine is added, 80~ 20~26h of microwave reaction is cooled to room temperature after the reaction was completed under conditions of 100 DEG C, methylene chloride is added, then wash 1~3 It is secondary, it is dry, it is recrystallized with dehydrated alcohol up to formula (IV) compound.

Formula (IV) compound is added to the in the mixed solvent of HCl- ethyl alcohol, wherein the concentration of HCl is 2~8mol/L, HCl Volume ratio with ethyl alcohol is 1:3~3:1, and outer temperature is heated to 100~120 DEG C of 10~12h of reflux, is cooled to room temperature, and white is precipitated Solid filters, and filter residue washs 1~3 time with dehydrated alcohol again, and obtained solid is added in dehydrated alcohol, and outer temperature is heated to 90~ 140 DEG C of 30~50min of reflux, are cooled to room temperature, and filter, and filter residue washs 1~3 time with dehydrated alcohol again to get formula (IV) chemical combination The hydrochloride of object.

Work as R₇When for OH, formula (IV) compound can be made by following steps:

Lavo-ofloxacin carboxylic acid is dissolved in DMSO, and TEA and (S/R) -7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazine is added, 20~26h of microwave reaction is cooled to room temperature after the reaction was completed under conditions of 80~100 DEG C, methylene chloride is added, then wash 1 It is~3 times, dry, it is recrystallized with dehydrated alcohol up to formula (IV) compound.

Formula (IV) compound is added to the in the mixed solvent of HCl- ethyl alcohol, wherein the concentration of HCl is 2~8mol/L, HCl Volume ratio with ethyl alcohol is 1:3~3:1, and outer temperature is heated to 100~120 DEG C of 10~12h of reflux, is cooled to room temperature, and white is precipitated Solid filters, and filter residue washs 1~3 time with dehydrated alcohol again, and obtained solid is added in dehydrated alcohol, and outer temperature is heated to 90~ 120 DEG C or more 30~50min of reflux, are cooled to room temperature, and filter, and filter residue washs 1~3 time with dehydrated alcohol again to get formula (IV) The hydrochloride of compound.

Work as R₈For 2,4-F₂C₆H₃(2,4- difluorophenyls,), R₉When for H, formula (V) compound and its hydrochloride can be with It is made by following steps:

The fluoro- Isosorbide-5-Nitrae of 1- (2,4- difluorophenyl) -6,7,8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid is dissolved in DMSO, is added Enter TEA and R- octahydro pyrrolo- [1,2-a] pyrazine, 10~14h of microwave reaction under conditions of 80~100 DEG C is cooled to room Temperature is added methylene chloride, then washes 1~3 time, dry, obtains formula (V) compound with dehydrated alcohol recrystallization；

Formula obtained (V) compound is added to the in the mixed solvent of HCl- ethyl alcohol, wherein the concentration of HCl is 2~8mol/ The volume ratio of L, HCl and ethyl alcohol is 1:3~3:1, and outer temperature is heated to 90~120 DEG C of 10~12h of reflux, is cooled to room temperature, and is precipitated White solid is filtered, then is washed 1~3 time with dehydrated alcohol；Obtained solid is added in dehydrated alcohol, and outer temperature is heated to 90~ 120 DEG C or more 30~50min of reflux, are cooled to room temperature, and filter, and filter residue is washed again with dehydrated alcohol to get formula (V) compound Hydrochloride.

Work as R₈For 2,4-F₂C₆H₃(2,4- difluorophenyls,), R₉When for OH, formula (V) compound and its hydrochloride can be by Following steps are made:

The fluoro- Isosorbide-5-Nitrae of 1- (2,4- difluorophenyl) -6,7,8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid is dissolved in DMSO, is added Enter TEA and R-7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazine, 10~14h of microwave reaction under conditions of 80~100 DEG C is cooling To room temperature, methylene chloride is added, then washes 1~3 time, it is dry, formula (V) compound is obtained with dehydrated alcohol recrystallization；

Work as R₈For c-C₃H₅(cyclopropyl,), R₉When for H, formula (V) compound and its hydrochloride can be by following steps systems :

By 1- cyclopropyl -6,7, the fluoro- Isosorbide-5-Nitrae of 8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid is dissolved in DMSO, be added TEA and R- octahydro pyrrolo- [1,2-a] pyrazine, 10~14h of microwave reaction under conditions of 80~100 DEG C are cooled to room temperature, and are added two Chloromethanes, then wash 1~3 time, it is dry, formula (V) compound is obtained with dehydrated alcohol recrystallization；

Work as R₈For c-C₃H₅(cyclopropyl,), R₉When for OH, formula (V) compound and its hydrochloride can be by following steps It is made:

By 1- cyclopropyl -6,7, the fluoro- Isosorbide-5-Nitrae of 8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid is dissolved in DMSO, be added TEA and S/R-7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazine, 10~14h of microwave reaction under conditions of 80~100 DEG C are cooled to room Temperature is added methylene chloride, then washes 1~3 time, dry, obtains formula (V) compound with dehydrated alcohol recrystallization；

Formula obtained (V) compound is added to the in the mixed solvent of HCl- ethyl alcohol, wherein the concentration of HCl is 2~8mol/L, The volume ratio of HCl and ethyl alcohol is 1:3~3:1, and outer temperature is heated to 90~120 DEG C of 10~12h of reflux, is cooled to room temperature, and is precipitated white Color solid is filtered, then is washed 1~3 time with dehydrated alcohol；Obtained solid is added in dehydrated alcohol, and outer temperature is heated to 90~120 DEG C or more 30~50min of reflux, be cooled to room temperature, filter, filter residue washs again with dehydrated alcohol to get formula (V) compound Hydrochloride.

Work as R₁₀For 2,4-F₂C₆H₃(2,4- difluorophenyls,), R₁₁When for H, formula (VI) compound and its hydrochloride can be by Following steps are made:

The fluoro- Isosorbide-5-Nitrae of 1- (2,4- difluorophenyl) -6,7,8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid is dissolved in DMSO, is added Enter TEA and S/R-7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazine, 10~14h of microwave reaction, cold under conditions of 80~100 DEG C But to room temperature, methylene chloride is added, then washes 1~3 time, it is dry, formula (VI) compound is obtained with dehydrated alcohol recrystallization；

Formula obtained (VI) compound is added to the in the mixed solvent of HCl- ethyl alcohol, wherein the concentration of HCl is 2~8mol/ The volume ratio of L, HCl and ethyl alcohol is 1:3~3:1, and outer temperature is heated to 90~120 DEG C of 10~12h of reflux, is cooled to room temperature, and is precipitated White solid is filtered, then is washed 1~3 time with dehydrated alcohol；Obtained solid is added in dehydrated alcohol, and outer temperature is heated to 90~ 120 DEG C or more 30~50min of reflux, are cooled to room temperature, and filter, and filter residue is washed again with dehydrated alcohol to get formula (VI) compound Hydrochloride.

Work as R₁₀For c-C₃H₅(cyclopropyl,), R₁₁When for H, formula (VI) compound and its hydrochloride can be by following steps It is made:

By 1- cyclopropyl -6,7, the fluoro- Isosorbide-5-Nitrae of 8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid is dissolved in DMSO, be added TEA and S/R- octahydro pyrrolo- [1,2-a] pyrazine, 10~14h of microwave reaction under conditions of 80~100 DEG C are cooled to room temperature, and are added Methylene chloride, then wash 1~3 time, it is dry, formula (VI) compound is obtained with dehydrated alcohol recrystallization；

Work as R₁₀For c-C₃H₅(cyclopropyl,), R₁₁When for OH, formula (VI) compound and its hydrochloride can be by following steps It is made:

By 1- cyclopropyl -6,7, the fluoro- Isosorbide-5-Nitrae of 8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid is dissolved in DMSO, be added TEA and S/R-7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazine, 10~14h of microwave reaction under conditions of 80~100 DEG C are cooled to room Temperature is added methylene chloride, then washes 1~3 time, dry, obtains formula (VI) compound with dehydrated alcohol recrystallization；

According to another aspect of the present invention, the present invention also provides formula (I), (II), (III), (IV), (V) or (VI) institutes The compound shown is preparing the application in resisting gram-positive bacteria and anti-Gram negative bacteria drugs, the drug can be tablet, Capsule, powder needle or injection type.

According to another aspect of the present invention, the present invention also provides formula (I), (II), (III), (IV), (V) or (VI) institutes The hydrochloride of the compound shown is preparing the application in resisting gram-positive bacteria and anti-Gram negative bacteria drugs, which can To be tablet, capsule, powder needle or injection type.

According to another aspect of the present invention, the present invention also provides for synthesize formula (I), (II), (III), (IV), (V) or the intermediate of (VI) compound, structure such as formula (VII), formula (VIII) and formula (Ⅸ) are shown.

Wherein, the R in formula (VII)₁₂It can be (S/R)-CH₂OH or (S/R)-CHOHCH₃；

R in formula (VIII)₁₃It can be H or OH；

R in formula (Ⅸ)₁₄It can be H or OH；

According to another aspect of the present invention, the present invention also provides the preparation methods of above-mentioned intermediate:

Work as R₁₂For (S/R)-CH₂When OH, formula (VII) compound can be made by following steps:

(S/R)-serine is added in anhydrous methanol, under ice salt bath, SOCl is slowly added dropwise₂, after being added dropwise, it is changed to room Temperature 6~15h of reaction, reaction are finished, and reaction solution is concentrated to dryness, and gained residue is dissolved in water, and under ice salt bath, NaHCO is added₃, to Gas release is finished, and ClCH is added dropwise₂COCl-CH₂Cl₂Solution, wherein ClCH₂COCl and CH₂Cl₂Volume ratio is 1:2~1:20, drop After adding, it is changed to 6~15h of room temperature reaction, after completion of the reaction, separates organic layer, CH₂Cl₂It aqueous layer extracted 1~3 time, is associated with Machine layer, is washed with saturated sodium chloride solution, and anhydrous sodium sulfate is added to dry, filter, evaporating solvent under reduced pressure, obtains weak yellow liquid (S/ R)-N- chloracetyl serine methylester.

N- chloracetyl serine methylester is dissolved in anhydrous methanol, and TEA and benzylamine is added in ice-water bath, outside 80~120 DEG C of temperature 40~56h of the above reflux, is cooled to room temperature, and decompression abstraction solvent, residue adds CH₂Cl₂Dissolution is washed 1~3 time, and chlorine is saturated Change sodium solution washing, the dry organic layer of anhydrous sodium sulfate, decompression abstraction CH₂Cl₂Crude product is obtained, column chromatographs to obtain white solid chirality (S/R) -1- benzyl -3- methylol -2,5- piperazinedione.

LiAlH₄It is suspended in anhydrous THF, under ice bath, compound chirality 1- benzyl -3- methylol -2,5- piperazine is added portionwise Piperazine diketone, finishes, and 30~50min, argon gas protection is stirred at room temperature, and 90~140 DEG C of outer temperature is heated to reflux 5~8h, and TLC detection is anti- It should finish, cold cause room temperature, H is successively added dropwise under ice bath₂O, concentration is NaOH, H of 2mol/L₂0.5~2h is stirred at room temperature in O, takes out Filter, CH₂Cl₂Washing, merging filtrate add anhydrous sulphur sodium to dry, filter, and abstraction solvent obtains yellow liquid (S/R) 1- benzyl -3- hydroxyl Methyl piperazine.

(S/R) -1- benzyl -3- methylol piperazine is dissolved in CH₂Cl₂In, add TEA, under ice bath, is added dropwise (Boc)₂O, drop finish, room Temperature reaction 10~14h, TLC detection, reaction are finished, and successively wash one with saturated sodium bicarbonate solution, water and saturated sodium chloride solution Secondary, organic layer adds anhydrous sodium sulfate dry, and rotation removes solvent, and column chromatography obtains yellow sticky matter (S/R) -1- tertbutyloxycarbonyl -2- Methylol -4- benzyl diethylenediamine.

(S/R) -1- tertbutyloxycarbonyl -2- methylol -4- benzyl diethylenediamine is dissolved in methanol, and Pd/C or/and Pd is added (OH)₂, glacial acetic acid 1~3 is added dropwise and drips, extracts air, is passed through H₂, 10~14h, filtering are reacted at room temperature, methanol washing merges filter Liquid, decompression extraction solvent obtain faint yellow solid formula (VII) compound (S/R) -1- tertbutyloxycarbonyl -2- methylol piperazine.

Work as R₁₂For (S/R)-CHOHCH₃When, formula (VII) compound can be made by following steps:

(S/R)-threonine is added in anhydrous methanol, under ice salt bath, SOCl is slowly added dropwise₂, after being added dropwise, it is changed to room Temperature 6~15h of reaction, reaction are finished, and reaction solution is concentrated to dryness, and gained residue is dissolved in water, and under ice salt bath, NaHCO is added₃, to Gas release is finished, and ClCH is added dropwise₂COCl-CH₂Cl₂Solution, wherein ClCH₂COCl and CH₂Cl₂Volume ratio is 1:2~1:20, drop After adding, it is changed to 6~15h of room temperature reaction, after completion of the reaction, separates organic layer, CH₂Cl₂It aqueous layer extracted 1~3 time, is associated with Machine layer, saturated sodium chloride solution washing, adds anhydrous sodium sulfate to dry, filter, evaporating solvent under reduced pressure obtains weak yellow liquid (S/ R)-N- chloracetyl threonine methyl.

N- chloracetyl threonine methyl is dissolved in anhydrous methanol, and TEA and benzylamine 80~120 DEG C of temperature outside is added in ice-water bath 40~56h of the above reflux, is cooled to room temperature, and decompression abstraction solvent, residue adds CH₂Cl₂Dissolution is washed 1~3 time, and chlorine is saturated Change sodium solution washing, the dry organic layer of anhydrous sodium sulfate, decompression abstraction CH₂Cl₂Crude product is obtained, column chromatographs to obtain white solid (S/ R) -1- benzyl -3- (1- ethoxy) -2,5- piperazinedione.

LiAlH₄It is suspended in anhydrous THF, under ice bath, compound 1- benzyl -3- (1- ethoxy) -2,5- piperazine is added portionwise Piperazine diketone, finishes, and 30~50min, argon gas protection is stirred at room temperature, and 90~140 DEG C of outer temperature is heated to reflux 5~8h, and TLC detection is anti- It should finish, cold cause room temperature, H is successively added dropwise under ice bath₂O, concentration is NaOH, H of 2mol/L₂0.5~2h is stirred at room temperature in O, takes out Filter, CH₂Cl₂Washing, merging filtrate add anhydrous sulphur sodium to dry, filter, and abstraction solvent obtains yellow liquid (S/R) -1- benzyl -3- (1- ethoxy) piperazine.

(S/R) -1- benzyl -3- (1- ethoxy) piperazine is dissolved in CH₂Cl₂In, add TEA, under ice bath, is added dropwise (Boc)₂O, drop Finish, react at room temperature 10~14h, TLC detection, reaction is finished, and saturated sodium bicarbonate solution, water and saturated sodium chloride solution are successively used It washed once, organic layer adds anhydrous sodium sulfate dry, and rotation removes solvent, and column chromatography obtains yellow sticky matter (S/R) -1- tertiary butyloxycarbonyl Base -2- (1- ethoxy) -4- benzyl diethylenediamine

(S/R) -1- tertbutyloxycarbonyl -2- (1- ethoxy) -4- benzyl diethylenediamine is dissolved in methanol, and Pd/C or/and Pd is added (OH)₂, glacial acetic acid 1~3 is added dropwise and drips, extracts air, is passed through H₂, 10~14h, filtering are reacted at room temperature, methanol washing merges filter Liquid, decompression extraction solvent obtain faint yellow solid formula (VII) compound chirality (S/R) -1- tertbutyloxycarbonyl -2- (1- ethoxy) piperazine Piperazine.

Work as R₁₃And R₁₄When being H, formula (VIII) and formula (Ⅸ) compound can be made by following steps:

(S/R)-proline is added in anhydrous methanol, under ice salt bath, SOCl is slowly added dropwise₂, after being added dropwise, it is changed to room Temperature 6~15h of reaction, reaction are finished, and reaction solution is concentrated to dryness, and gained residue is dissolved in water, and under ice salt bath, NaHCO is added₃, to Gas release is finished, and ClCH is added dropwise₂COCl-CH₂Cl₂Solution, wherein ClCH₂COCl and CH₂Cl₂Volume ratio is 1:2~1:20, drop After adding, it is changed to 6~15h of room temperature reaction, after completion of the reaction, separates organic layer, CH₂Cl₂It aqueous layer extracted 1~3 time, is associated with Machine layer, saturated sodium chloride solution washing, adds anhydrous sodium sulfate to dry, filter, evaporating solvent under reduced pressure obtains yellow oil (S/ R)-N- chloracetyl proline methyl ester.

(S/R)-N- chloracetyl proline methyl ester is dissolved in anhydrous methanol, ice-water bath, be added TEA and benzylamine outside temperature 80~ 120 DEG C of 40~56h of reflux, are cooled to room temperature, and decompression abstraction solvent, residue adds CH₂Cl₂Dissolution is washed 1~3 time, and chlorine is saturated Change sodium solution washing, the dry organic layer of anhydrous sodium sulfate, decompression abstraction CH₂Cl₂Crude product is obtained, column chromatographs to obtain white solid (S/ R) -2- benzyl-hexahydropyrrolo simultaneously [1,2-a] pyrazine-Isosorbide-5-Nitrae diketone.

LiAlH4 is suspended in anhydrous THF, under ice bath, be added portionwise compound (S/R) -2- benzyl-hexahydropyrrolo simultaneously [1, 2-a] pyrazine-Isosorbide-5-Nitrae diketone, it finishes, 30~50min, argon gas protection is stirred at room temperature, 90~140 DEG C of outer temperature is heated to reflux 5~8h, TLC detects end of reaction, and H is successively added dropwise under ice bath in cold cause room temperature₂O, concentration is NaOH, H2O of 2mol/L, is stirred at room temperature 0.5~2h is filtered, CH₂Cl₂Washing, merging filtrate add anhydrous sulphur sodium to dry, filter, and abstraction solvent obtains (S/R) -2- benzyl eight Hydrogen pyrrolo- [1,2-a] pyrazine.

(S/R) -2- benzyl octahydro pyrrolo- [1,2-a] pyrazine is dissolved in methanol, and Pd/C or Pd (OH) is added₂, ice is added dropwise Acetic acid 1~3 drips, and extracts air, is passed through H₂, 10~14h is reacted at room temperature, filtering, methanol washing, merging filtrate, decompression is extracted out molten Agent obtains weak yellow liquid formula (VIII) or formula (Ⅸ) compound (S/R) octahydro pyrrolo- [1,2-a] pyrazine.Wherein from R- proline It sets out obtained formula (VIII) compound for raw material, is the obtained formula of raw material (Ⅸ) compound from S- proline.

Work as R₁₃And R₁₄When being OH, formula (VIII) and formula (Ⅸ) compound can be made by following steps:

(S/R) -4- hydroxy-proline is added in anhydrous methanol, under ice salt bath, SOCl is slowly added dropwise₂, after being added dropwise, It is changed to 6~15h of room temperature reaction, reaction is finished, and reaction solution is concentrated to dryness, and gained residue is dissolved in water, and under ice salt bath, is added NaHCO₃, finish to gas release, ClCH be added dropwise₂COCl-CH₂Cl₂Solution, wherein ClCH₂COCl and CH₂Cl₂Volume ratio is 1:2 ~1:20 after being added dropwise, is changed to 6~15h of room temperature reaction, after completion of the reaction, separates organic layer, CH₂Cl₂Aqueous layer extracted 1~3 It is secondary, merge organic layer, saturated sodium chloride solution washing adds anhydrous sodium sulfate to dry, filter, evaporating solvent under reduced pressure obtains yellow oil Shape object chirality (S/R)-N- chloracetyl -4- L-Hydroxyproline methyl ester.

(S/R)-N- chloracetyl -4- L-Hydroxyproline methyl ester is dissolved in anhydrous methanol, and ice-water bath, is added TEA and benzylamine exists 80~120 DEG C of 40~56h of reflux of outer temperature, are cooled to room temperature, and decompression abstraction solvent, residue adds CH₂Cl₂Dissolution, washing 1~3 It is secondary, saturated sodium chloride solution washing, the dry organic layer of anhydrous sodium sulfate, decompression abstraction CH₂Cl₂Crude product is obtained, column chromatographs white Color Solid Chiral (S/R) -2- benzyl -7- hydroxyl hexahydropyrrolo simultaneously [1,2-a] pyrazine-Isosorbide-5-Nitrae diketone.

LiAlH₄It is suspended in anhydrous THF, under ice bath, compound chirality (S/R) -2- benzyl -7- hydroxyl six is added portionwise Hydrogen pyrrolo- [1,2-a] pyrazine-Isosorbide-5-Nitrae diketone, finishes, and 30min, argon gas protection is stirred at room temperature, and 90~140 DEG C of outer temperature heats back 5~8h is flowed, TLC detects end of reaction, and H is successively added dropwise under ice bath in cold cause room temperature₂O、2N NaOH、H₂O, it is stirred at room temperature 0.5~ 2h is filtered, CH₂Cl₂Washing, merging filtrate add anhydrous sulphur sodium to dry, filter, and abstraction solvent obtains (S/R) -2- benzyl -7- hydroxyl Octahydro pyrrolo- [1,2-a] pyrazine.

(S/R) -2- benzyl -7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazine is dissolved in methanol, and Pd/C or Pd (OH) is added₂, Glacial acetic acid 1~3 is added dropwise to drip, extracts air, is passed through H₂, react at room temperature 10~14h, filtering, methanol washing, merging filtrate, decompression Extraction solvent obtains weak yellow liquid formula (VIII) or formula (Ⅸ) compound (S/R) -7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazine.Its In from R-4- hydroxy-proline be the obtained formula of raw material (VIII) compound, from S-4- hydroxy-proline be raw material it is obtained Formula (Ⅸ) compound.

Detailed description of the invention

Fig. 1 is the hydrogen nuclear magnetic resonance spectrogram of compound S1.

Fig. 2 is the carbon-13 nmr spectra figure of compound S1.

Fig. 3 is the hydrogen nuclear magnetic resonance spectrogram of compound S2.

Fig. 4 is the carbon-13 nmr spectra figure of compound S2.

Fig. 5 is the hydrogen nuclear magnetic resonance spectrogram of compound S3.

Fig. 6 is the carbon-13 nmr spectra figure of compound S3.

Fig. 7 is the hydrogen nuclear magnetic resonance spectrogram of compound S4.

Fig. 8 is the carbon-13 nmr spectra figure of compound S4.

Fig. 9 is the hydrogen nuclear magnetic resonance spectrogram of compound S5.

Figure 10 is the carbon-13 nmr spectra figure of compound S5.

Figure 11 is the hydrogen nuclear magnetic resonance spectrogram of compound s 6.

Figure 12 is the carbon-13 nmr spectra figure of compound s 6.

Figure 13 is the hydrogen nuclear magnetic resonance spectrogram of compound S7.

Figure 14 is the carbon-13 nmr spectra figure of compound S7.

Figure 15 is the hydrogen nuclear magnetic resonance spectrogram of compound S8.

Figure 16 is the carbon-13 nmr spectra figure of compound S8.

Figure 17 is the hydrogen nuclear magnetic resonance spectrogram of compound S9.

Figure 18 is the carbon-13 nmr spectra figure of compound S9.

Figure 19 is the hydrogen nuclear magnetic resonance spectrogram of compound S10.

Figure 20 is the carbon-13 nmr spectra figure of compound S10.

Figure 21 is the hydrogen nuclear magnetic resonance spectrogram of compound S11.

Figure 22 is the carbon-13 nmr spectra figure of compound S11.

Figure 23 is the hydrogen nuclear magnetic resonance spectrogram of compound S12.

Figure 24 is the carbon-13 nmr spectra figure of compound S12.

Figure 25 is the hydrogen nuclear magnetic resonance spectrogram of compound S1-TM.

Figure 26 is the hydrogen nuclear magnetic resonance spectrogram of compound S2-TM.

Figure 27 is the hydrogen nuclear magnetic resonance spectrogram of compound S3-TM.

Figure 28 is the hydrogen nuclear magnetic resonance spectrogram of compound S4-TM.

Figure 29 is the hydrogen nuclear magnetic resonance spectrogram of compound S5-TM.

Figure 30 is the hydrogen nuclear magnetic resonance spectrogram of compound s 6-TM.

Figure 31 is the hydrogen nuclear magnetic resonance spectrogram of compound R 1-TM.

Figure 32 is the hydrogen nuclear magnetic resonance spectrogram of compound R 2-TM.

Figure 33 is the hydrogen nuclear magnetic resonance spectrogram of compound R 3-TM.

Figure 34 is the hydrogen nuclear magnetic resonance spectrogram of compound R 4-TM.

Figure 35 is the hydrogen nuclear magnetic resonance spectrogram of compound R 5-TM.

Figure 36 is the hydrogen nuclear magnetic resonance spectrogram of compound R 6-TM.

Specific embodiment

Below by embodiment, the present invention is described in further detail.

Embodiment 1

(S) the fluoro- 2,3- dihydro -3- methyl-1 0- of -9- (4- tertbutyloxycarbonyl -3- methylol -1- piperazinyl) -7- oxo - (3S) -7H- pyrido [1,2,3-de]-Isosorbide-5-Nitrae-benzoxazine -6- carboxylic acid (for formula (II) structure, hereinafter referred to as compound S1) and The synthesis of its hydrochloride (hereinafter referred to as compound S1-TM).

Serine (10.50g, 0.10mol) is added in anhydrous methanol (200mL), under ice salt bath, SOCl is added dropwise₂ (29.01mL, 0.40mol) is changed to react at room temperature after being added dropwise, and TLC tracing detection is dense by reaction solution to after completion of the reaction It is reduced to dry, gained residue (serine methyl ester hydrochloride, hereinafter referred to as compound S1-a) directly carries out the next step.

Residue adds water (120mL), and under ice salt bath, NaHCO is added₃(19.75g, 0.24mol) finishes, drop to gas release Add ClCH₂The CH of COCl (8.0mL, 0.11mol)₂Cl₂(120mL) solution, 30min drop finish, and it is anti-to be changed to room temperature after being added dropwise It answers, TLC tracing detection to end of reaction separates organic layer, with CH₂Cl₂Multiple aqueous layer extracted merges organic layer, adds anhydrous sulphur Sour sodium dries, filters, and 30 DEG C of rotary evaporations remove solvent, and obtaining light yellow liquid, (N- chloracetyl serine methylester, hereinafter referred to as changes Close object S1-1) 11.17g.

Compound S1-1 (9.78g, 0.05mol) is dissolved in anhydrous methanol (100mL), ice-water bath sequentially adds TEA (20.6mL, 0.15mol) and benzylamine (6.7mL, 0.06mol), flow back 48h under conditions of 80 DEG C, is cooled to room temperature, and is added A large amount of white solids are precipitated in 30mL ethyl acetate, freezing, filter, then wash filter cake 3 times (each 20mL) with ethyl acetate, very White solid 4.36g is obtained after sky is dry.Mother liquor rotary evaporation removes solvent, faint yellow solid is obtained, with CH₂Cl₂(30mL) dissolution, Column chromatographs to obtain white solid 4.32g, merging be obtained white solid (1- benzyl -3- methylol -2,5- piperazinedione, hereinafter referred to as Compound S1-2) 8.68g.

By LiAlH₄(2.88g, 76mmol) is suspended in anhydrous THF (tetrahydrofuran, 60mL), under ice bath, portion-wise with caution It is added compound S1-2 (4.69g, 20mmol), finishes, 30min is stirred at room temperature, lower outer 90 DEG C of the temperature of argon gas protection is heated to reflux 4h, TLC detect end of reaction, are cooled to room temperature, H is successively added dropwise under ice bath₂O (2.8mL), NaOH (concentration 2N, 9.0mL), H₂O (2.8mL), is stirred at room temperature 1h, filters, then use CH₂Cl₂3 times (each 10mL) is washed, merging filtrate adds anhydrous sodium sulfate It dries, filters, filters and remove solid, obtain 4.06g yellow liquid (1- benzyl -3- methylol piperazine, hereinafter referred to as compound S1- 3).Refrigeration transforms into semisolid, is easy to the moisture absorption in air.

Compound S1-3 (2.06g, 10mmol) is taken to be dissolved in 15mLCH₂Cl₂In, add TEA (1.53mL, 11mmol), ice bath Under, it is added dropwise (Boc)₂O (2.40g, 11mmol), is added dropwise, and reacts at room temperature 12h, and TLC detects end of reaction, successively uses 10mL Saturated sodium bicarbonate solution, 10mL water, 10mL saturated sodium chloride solution are respectively washed 1 time, and organic layer adds anhydrous sodium sulfate dry, Rotary evaporation remove solvent, column chromatography, obtain yellow sticky matter (1- tertbutyloxycarbonyl -2- methylol -4- benzyl diethylenediamine, hereinafter referred to as Compound S1-4) 2.68g.

It takes compound S1-4 (2.00g, 6.5mmol) to be dissolved in 20mL methanol, is added Pd/C (0.20g), Pd (OH)₂ (0.10g) is added dropwise glacial acetic acid 1 and drips, extract air, be passed through H₂, react at room temperature 12h.Filtering, again with methanol wash 3 times (every time 5mL), merging filtrate is spin-dried for solvent and obtains faint yellow solid (1- tertbutyloxycarbonyl -2- methylol piperazine, hereinafter referred to as compound S1- 5)1.38g。

Lavo-ofloxacin carboxylic acid (225.0mg, 0.8mmol) is dissolved in DMSO (dimethyl sulfoxide, 5.0mL), TEA is added (triethanolamine, 0.44mL, 3.2mmol) and compound S1-5 (259.5mg, 1.2mmol), microwave is anti-under conditions of 90 DEG C It should for 24 hours.After the reaction was completed, it is cooled to room temperature, is added methylene chloride (15mL), then wash 3 times (each 15mL), it is dry, with nothing Water-ethanol recrystallizes to obtain yellow solid 105.1mg, calculate yield be 27.51%, measure mp (fusing point): 221-223 DEG C.

The raw material utilized in the above operation is Serine, acquisition be S configuration product.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy, carbon-13 nmr spectra are as shown in Figure 1 and Figure 2, and specific spectral data is as follows, thus really Fixed output quota object is compound S1.

¹H NMR (500MHz, DMSO) δ 15.17 (d, J=3.5Hz, 1H), 8.97 (d, J=1.5Hz, 1H), 7.57- 7.60 (m, 1H), 4.91 (d, J=6.5Hz, 1H), 4.68-4.72 (m, 1H), 4.56-4.58 (m, 1H), 4.34-4.39 (m, 1H), 3.99 (s, 1H), 3.81 (d, J=10.5Hz, 1H), 3.69-3.76 (m, 1H), 3.45-3.52 (m, 2H), 3.23- 3.26(m,1H),3.11-3.18(m,2H),1.42-1.46(m, 12H)；

¹³C NMR(126MHz,DMSO)δ176.36,165.99,155.59,154.26,146.17,140.70,132.04, 124.75,120.07, 106.72,103.15,78.86,68.08,57.87,54.83,52.87,49.9,49.5,28.04, 17.87；

HRMS-ESI(m/z):Calcd.For C₂₃H₂₈FN₃NaO₇(M+Na)⁺:500.18090；Found:500.17975.

Compound S1 (29.1mg, 0.061mmol) is added to HCl (concentration 3N, 2mL)-ethyl alcohol (5mL) in the mixed solvent, 95 DEG C of outer temperature is heated to reflux 10h, is cooled to room temperature, and white solid is precipitated, and filters, and filter residue washs 3 times (often with dehydrated alcohol again Secondary 1mL).Obtained solid is added in 5mL dehydrated alcohol, and 95 DEG C of outer temperature is heated to reflux 30min, is cooled to room temperature, and is filtered, filter residue Washed 3 times (each 1mL) with dehydrated alcohol again, obtain yellow solid 13.2mg, calculate yield be 52.29%, measure mp > 250℃。

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy is as shown in figure 25, and specific spectral data is as follows, thereby determines that product is compound S1- TM。

¹H NMR (400MHz, DMSO) δ 15.10 (s, 1H), 9.54 (s, 1H), 9.20 (s, 1H), 8.99 (d, J= 5.6Hz, 1H), 7.59-7.64 (m, 1H), 5.47 (s, 1H), 4.94 (s, 1H), 4.61 (d, J=8.4Hz, 1H), 4.40 (d, J =11.2Hz, 1H), 3.68 (s, 2H), 3.39-3.50 (m, 4H), 3.13 (s, 1H), 1.46 (s, 3H)；

HRMS-ESI(m/z):Calcd.For C₁₈H₂₁FN₃O₅₍M+H)⁺:378.14653；Found:378.14 553.

Embodiment 2

(S) the fluoro- 2,3- dihydro -3- methyl-1 0- of -9- (4- tertbutyloxycarbonyl -3- (1- ethoxy) -1- piperazinyl) -7- oxygen Generation-(3S) -7H- pyrido [1,2,3-de]-Isosorbide-5-Nitrae-benzoxazine -6- carboxylic acid (for formula (II) structure, hereinafter referred to as compound S2) And its synthesis of hydrochloride (hereinafter referred to as compound S2-TM).

Threonine (11.95g, 0.10mol) is added in anhydrous methanol (200mL), under ice salt bath, SOCl is added dropwise₂ (29.01mL, 0.40mol), is added dropwise, and is changed to react at room temperature, and reaction solution is concentrated by TLC tracing detection to end of reaction Dry, gained residue (threonine methyl ester hydrochloric salt, hereinafter referred to as compound S2-a) directly carries out the next step.

Gained residue adds water (120mL), and under ice salt bath, NaHCO is added₃(19.75g, 0.24mol) is discharged to gas Finish, ClCH is added dropwise₂The CH of COCl (8.0mL, 0.11mol)₂Cl₂(120mL) solution, 30min are added dropwise, and it is anti-to be changed to room temperature It answers, reaction is finished, and organic layer, CH are separated₂Cl₂Multiple aqueous layer extracted.Merge organic layer, anhydrous sodium sulfate is added to dry, filter, 30 DEG C Decompression, which filters, removes solvent, obtains faint yellow solid (N- chloracetyl threonine methyl, hereinafter referred to as compound S2-1) 17.05g.

Compound S2-1 (14.67g, 70mmol) is dissolved in anhydrous methanol (140mL), ice-water bath sequentially adds TEA (28.8mL, 0.21mol) and benzylamine (9.34mL, 85.4mmol), 80 DEG C of reflux 48h of outer temperature, is cooled to room temperature, and is precipitated a large amount of White solid filters, then washs filter cake 3 times (each 20mL) with ethyl acetate, and white solid 10.96g is obtained after vacuum drying. Mother liquor depressurizes abstraction solvent again, obtains yellow solid, and column chromatographs to obtain white solid 1.18g, and white solid (1- benzyl -3- is obtained (1- ethoxy) -2,5- piperazinedione, hereinafter referred to as compound S2-2) 12.14g.

By LiAlH₄(2.88g, 76mmol) is suspended in anhydrous THF (60mL), and under ice bath, compound is added in portion-wise with caution S2-2 (4.97g, 20mmol), finishes, and 30min is stirred at room temperature, and lower outer 90 DEG C of the temperature of argon gas protection is heated to reflux 4h, and TLC detection is anti- It should finish, be cooled to room temperature, H is successively added dropwise under ice bath₂O (2.8mL), NaOH (concentration 2N, 9.0mL), H₂O (2.8mL), room temperature 1h is stirred, is filtered, then use CH₂Cl₂3 times (each 10mL) is washed, merging filtrate adds anhydrous sodium sulfate to dry, filter, and decompression is taken out Except solvent obtains 4.63g yellow liquid (1- benzyl -3- (1- ethoxy) piperazine, hereinafter referred to as compound S2-3).

Compound S2-3 (2.20g, 10mmol) is dissolved in the CH of 15mL₂Cl₂In, it is added TEA (1.53mL, 11mmol), ice Under bath, it is added dropwise (Boc)₂O (2.40g, 11mmol) after being added dropwise, reacts at room temperature 12h, and TLC detects end of reaction, successively uses 10mL saturated sodium bicarbonate solution, 10mL water, 10mL saturated sodium chloride solution are respectively washed 1 time, and organic layer adds anhydrous sodium sulfate dry It is dry, decompression abstraction solvent, column chromatography, obtain yellow sticky matter (1- tertbutyloxycarbonyl -2- (1- ethoxy) -4- benzyl diethylenediamine, with Call compound S2-4 in the following text) 2.70g.

It takes compound S2-4 (2.15g, 6.7mmol) to be dissolved in 15mL methanol, Pd/C (0.22g) and Pd (OH) is added₂ (0.10g) is added dropwise glacial acetic acid 1 and drips, extract air, be passed through H₂, 12h, filtering are reacted at room temperature, again with methanol washs 3 times (every time 5mL), merging filtrate, decompression filter remove solvent obtain faint yellow solid (1- tertbutyloxycarbonyl -2- (1- ethoxy) piperazine, with Call compound S2-5 in the following text) 1.54g.

Referring to the synthetic method of compound S1 in embodiment 1, by compound S1-5 be substituted for compound S2-5 (276.4mg, 1.2mmol) with lefofloxacin carboxylic acid reaction.Yellow solid 78.5mg, calculate yield be 19.96%, measure mp:210-212℃。

The raw material utilized in the above operation is L-threonine, acquisition be S configuration product.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy, carbon-13 nmr spectra are as shown in Figure 3, Figure 4, and specific spectral data is as follows, thus really Fixed output quota object is compound S2.

¹H NMR (500MHz, DMSO) δ 15.12 (s, 1H), 8.98 (s, 1H), 7.58 (d, J=12.0Hz, 1H), 4.92- 4.94 (m, 1H), 4.51-4.57 (m, 2H), 4.39 (dd, J=11.4,2.1Hz, 1H), 4.23-4.27 (m, 1H), 3.85 (d, J=10.0Hz, 1H), 3.77 (s, 1H), 3.25-3.29 (m, 2H), 3.19-3.22 (m, 3H), 1.44 (d, J=7.0Hz, 3H), 1.41 (s, 9H), 1.15 (d, J=6.5Hz, 3H)；

¹³C NMR(126MHz,DMSO)δ176.77,166.37,156.44,155.39,146.61,141.66,132.22, 125.08,121.04, 107.25,103.64,78.89,68.60,63.28,57.85,55.25,51.53,50.60,28.55, 21.79,18.20；

HRMS-ESI(m/z):Calcd.For C₂₄H₃₀FN₃NaO₇(M+Na)⁺:514.19655；Found:514.19542.

Referring to the synthetic method of S1-TM in embodiment 1, compound S1 is replaced with into compound S2 (49.1mg, 0.1mmol) Reacted, obtain yellow solid 31.1mg, calculate yield be 72.50%, measure 250 DEG C of mp >.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy is as shown in figure 26, and specific spectral data is as follows, thereby determines that product is compound S2- TM。

¹H NMR (400MHz, DMSO) δ 15.11 (s, 1H), 9.21 (s, 1H), 9.00-9.05 (m, 2H), 7.62 (d, J= 12.0Hz, 2H), 5.65 (s, 1H), 4.94 (d, J=6.8Hz, 1H), 4.60 (d, J=11.6Hz, 1H), 4.41 (d, J= 11.2Hz, 1H), 3.85 (t, J=13.2Hz, 1H), 3.25-3.53 (m, 5H), 3.09 (s, 2H), 1.45 (d, J=6.8Hz, 3H), 1.18 (d, J=6.4Hz, 3H)；

HRMS-ESI(m/z):Calcd.For C₁₉H₂₃FN₃O₅(M+H)⁺:392.16217；Found:392.16138.

Embodiment 3

(S) the fluoro- 2,3- dihydro -3- methyl-1 0- of -9- (octahydro pyrrolo- [1,2-a] pyrazinyl) -7- oxo-(3S) -7H- The synthesis of pyrido [1,2,3-de]-Isosorbide-5-Nitrae-benzoxazine -6- carboxylic acid (for formula (IV) structure, hereinafter referred to as compound S3).

Proline (11.51g, 0.10mol) is added in anhydrous methanol (200mL), under ice salt bath, SOCl is added dropwise₂ (29.01mL, 0.40mol), is added dropwise, and is changed to react at room temperature, and TLC tracing detection to end of reaction, reaction solution is concentrated into Dry, gained residue (proline methyl ester hydrochloride, hereinafter referred to as compound S3-a) directly carries out the next step.

Up add water (80mL) in the resulting residue of step, under ice salt bath, NaHCO is added₃(19.74g, 0.24mol), Finish to gas release, ClCH is added dropwise₂The CH of COCl (9.50mL, 0.12mol)₂Cl₂(50mL) solution, 30min are added dropwise, change To react at room temperature 6h, organic layer is separated after completion of the reaction, then use CH₂Cl₂Aqueous layer extracted 3 times (each 20mL), merge organic layer, Washed with saturated sodium chloride solution (30mL), then plus anhydrous sodium sulfate dry, filter, vacuum distillation remove solvent, obtain yellow oil Shape object (N- chloracetyl proline methyl ester, hereinafter referred to as compound S3-1) 11.64g.

Compound S3-1 (11.34g, 55mmol) is dissolved in anhydrous methanol (120mL), ice-water bath, addition TEA (9.15ml, It 66mmol) is cooled to room temperature with benzylamine (7.07g, 66mmol), 80 DEG C of reflux 48h of outer temperature, decompression, which filters, removes solvent, filter residue Add CH₂Cl₂(100mL) dissolution, is first washed with water 2 times (each 60mL), is then washed with saturated sodium chloride solution (30mL), finally With the dry organic layer of anhydrous sodium sulfate, decompression, which filters, removes CH₂Cl₂Crude product is obtained, column chromatographs to obtain white solid (2- benzyl-six Hydrogen pyrrolo- [1,2-a] pyrazine-Isosorbide-5-Nitrae diketone, hereinafter referred to as compound S3-2) 11.15g.

By LiAlH₄(4.33g, 114mmol) is suspended in anhydrous THF (100mL), and under ice bath, compound is added portionwise After adding, 30min is stirred at room temperature in S3-2 (7.33g, 30mmol), and lower outer 90 DEG C of the temperature of argon gas protection is heated to reflux 5h, TLC detection End of reaction is cooled to room temperature, and H is successively added dropwise under ice bath₂O (8.4mL), NaOH (concentration 2N, 27.0mL), H₂O 1h is stirred at room temperature in (8.4mL), filters, filter residue CH₂Cl₂It is filtered again after washing, merging filtrate simultaneously adds anhydrous sulphur sodium dry, takes out It filters out solvent and obtains 6.42g liquid (2- benzyl octahydro pyrrolo- [1,2-a] pyrazine, hereinafter referred to as compound S3-3).

Compound S3-3 (4.33g, 20mmol) is dissolved in methanol (50mL), is added Pd/C (0.45g), Pd (OH)₂ (0.20g) is added dropwise glacial acetic acid 1 and drips, extract air, be passed through H₂, 12h, filtering are reacted at room temperature, filter residue washs 3 times (often with methanol Secondary 5mL), merging filtrate, decompression suction filtration removing solvent obtains weak yellow liquid, and (octahydro pyrrolo- [1,2-a] pyrazine, hereinafter referred to as changes Close object S3-4) 1.83g.

Referring to the synthetic method of compound S1 in embodiment 1, by compound S1-5 be substituted for compound S3-4 (151.4mg, 1.2mmol) with lefofloxacin carboxylic acid reaction, obtain yellow solid 147.7mg, calculate yield be 47.66%, survey Obtain mp:237-240 DEG C.

The raw material utilized in the above operation is L-PROLINE, acquisition be S configuration product.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy, carbon-13 nmr spectra are as shown in Figure 5, Figure 6, thereby determine that product is compound S3.

¹H NMR (500MHz, DMSO) δ 15.19 (s, 1H), 8.95 (s, 1H), 7.54 (d, J=12.5Hz, 1H), 4.91 (q, J=6.5Hz, 1H), 4.57-4.95 (m, 1H), 4.33-4.37 (m, 1H), 3.46 (d, J=10.5Hz, 1H), 3.32 (d, J=7.0Hz, 1H), 3.22 (t, J=11.0Hz, 1H), 2.97-3.03 (m, 3H), 2.25-2.30 (m, 1H), 2.00-2.14 (m, 2H), 1.63-1.78 (m, 3H), 1.45 (d, J=6.5Hz, 3H), 1.27-1.35 (m, 1H)；

¹³C NMR(126MHz,DMSO)δ176.82,166.49,155.91,146.55,140.54,132.77,125.24, 120.00,107.09, 103.69,68.53,62.87,55.68,55.30,53.59,52.37,50.04,27.39,21.22, 18.33；

HRMS-ESI(m/z):Calcd.For C₂₀H₂₃FN₃O₄(M+H)⁺:388.16726；Found:388.16545.

Embodiment 4

(S) the fluoro- 2,3- dihydro -3- methyl-1 0- of -9- (7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazinyl) -7- oxo - (3S) -7H- pyrido [1,2,3-de]-Isosorbide-5-Nitrae-benzoxazine -6- carboxylic acid (for formula (IV) structure, hereinafter referred to as compound S4) Synthesis.

4- hydroxy-proline (19.67g, 0.15mol) is added in anhydrous methanol (300mL), under ice salt bath, is added dropwise SOCl₂(43.8 mL, 0.60mol), are added dropwise, and are changed to react at room temperature, and TLC tracing detection is to after completion of the reaction by reaction solution It is concentrated to dryness, it is anti-that gained residue (4- L-Hydroxyproline methyl ester hydrochloride, hereinafter referred to as compound S4-a) directly carries out lower step It answers.

Resulting residue adds water (240mL), and under ice salt bath, NaHCO is added₃(29.61g, 0.35mol), is released to gas Put complete, dropwise addition ClCH₂The CH of COCl (12.42mL, 0.17mol)₂Cl₂(180mL) solution, 30min are added dropwise, and are changed to room temperature 6h is reacted, organic layer is separated after completion of the reaction, uses CH₂Cl₂Aqueous layer extracted 3 times (each 20mL), merges organic layer, add anhydrous sulphur Sour sodium dries, filters, vacuum distillation remove solvent, obtain yellow oil (N- chloracetyl -4- L-Hydroxyproline methyl ester, below Claim compound S4-1) 25.26g.

Compound S4-1 (22.16g, 0.10mol) is dissolved in anhydrous methanol (200mL), under ice-water bath, TEA is added (16.63mL, 0.12mol) and benzylamine (11.79g, 0.12mol), 80 DEG C of reflux 48h of outer temperature, is cooled to room temperature, and decompression filters Solvent is removed, residue adds CH₂Cl₂(100mL) makes it dissolve, and column chromatography obtains white solid (2- benzyl -7- hydroxyl hexahydro pyrrole Cough up simultaneously [1,2- α] pyrazine-Isosorbide-5-Nitrae diketone, hereinafter referred to as compound S4-2) 21.5g.

Take LiAlH₄(1.71g, 45mmol) is suspended in anhydrous THF (40mL), and under ice bath, compound S4-2 is added portionwise (3.90g, 15mmol), is stirred at room temperature 30min after adding, lower outer 90 DEG C of the temperature of argon gas protection is heated to reflux 5h, TLC detection reaction It finishes, is cooled to room temperature, H is successively added dropwise under ice bath₂O(2.1mL)、2N NaOH(9.0mL)、H₂O (2.1mL), magnetic agitation 1h is filtered, filter residue CH₂Cl₂3 times (each 10mL) is washed, merging filtrate is added anhydrous sodium sulfate and dries, filters, and suction filtration removes Solvent is gone to obtain crude product, column chromatographs to obtain 1.94g liquid, and standing solidifies to obtain faint yellow solid (2- benzyl -7- hydroxyl octahydro pyrroles And [1,2-a] pyrazine, hereinafter referred to as compound S4-3).

Compound S4-3 (1.80g, 7.74mmol) is dissolved in 15mL methanol, Pd/C (0.18g) and Pd (OH) is added₂ (0.10g) is added dropwise glacial acetic acid 1 and drips, extract air, be passed through H₂, react at room temperature 12h.Filtering, filter residue wash 3 times (often with methanol Secondary 5mL), merging filtrate, decompression filter remove solvent obtain faint yellow solid (7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazine, with Call compound S4-4 in the following text) 0.98g.

Referring to the synthetic method of compound S1 in embodiment 1, compound S1-5 is substituted for S4-4 and lefofloxacin Carboxylic acid reaction.Lavo-ofloxacin carboxylic acid (112.5mg, 0.4mmol) is dissolved in DMSO (3.0mL), TEA is added (0.22mL, 1.6mmol) and compound S4-4 (85.3mg, 0.6mmol), microwave reaction is for 24 hours, cooling under conditions of 90 DEG C To room temperature, be added methylene chloride (15mL), then wash 3 times (each 15mL), it is dry, with dehydrated alcohol recrystallize yellow is solid Body 63.5mg, yield 39.35%, measures mp:229-232 DEG C.

The raw material utilized in the above operation is L-4- hydroxy-proline, acquisition be S configuration product.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy, carbon-13 nmr spectra, mass spectrogram are as shown in Figure 7, Figure 8, and specific spectral data is as follows, Thereby determine that product is compound S4.

¹H NMR (500MHz, DMSO) δ 15.21 (s, 1H), 8.95 (s, 1H), 7.56 (d, J=12.5Hz, 1H), 4.91 (d, J=7.0Hz, 1H), 4.78 (d, J=4.5Hz, 1H), 4.57-4.59 (m, 1H), 4.35-4.37 (m, 1H), 4.22 (s, 1H), 3.44 (d, J=12.0Hz, 1H), 3.19 (t, J=11.0Hz, 1H), 2.90-2.96 (m, 2H), 2.32-2.38 (m, 2H), 2.01 (dd, J=9.0,5.5Hz, 1H), 1.56-1.60 (m, 2H), 1.45 (d, J=6.5Hz, 3H)；

¹³C NMR(126MHz,DMSO)δ176.82,166.51,155.90,146.55,140.56,132.72,125.25, 120.01, 107.09,103.70,68.52,68.02,63.42,61.07,55.49,55.30,52.16,50.14,39.05, 18.34；

HRMS-ESI(m/z):Calcd.For C₂₀H₂₃FN₃O₅(M+H)⁺:404.16218；Found:404.16134.

Embodiment 5

(S) the fluoro- 7- of -1- (2,4 difluorobenzene base) -6,8- two (4- tertbutyloxycarbonyl -3- methylol -1- piperazinyl) -1,4- Dihydro -4- Oxoquinoline-3-carboxylic acid (for formula (I) structure, hereinafter referred to as compound S5) and its hydrochloride (hereinafter referred to as compound S3- TM synthesis).

2,3,4,5- tetrafluorobenzoyl ethyl (11.89g, 45mmol) is added into triethyl orthoformate In (11.24mL, 67.5mmol) and aceticanhydride (20mL), 140 DEG C of reaction 1h are warming up to, vacuum distillation removing solvent obtains faint yellow Liquid.Gained weak yellow liquid is dissolved in methylene chloride (150mL), under protection of argon gas, is cooled with an ice bath, is added 2,4- Difluoroaniline (6.87mL, 67.5mmol) reacts at room temperature 1h, and decompression rotation obtains yellow solid except solvent, and petroleum ether recrystallizes white Color solid (2- (2,3,4,5- tetrafluoro benzoyl) -3- (2,4- difluorophenyl)-ethyl acrylate, hereinafter referred to as compound MH3- 1)15.44g。

Compound MH3-1 (14.92g, 37mmol) is dissolved in the DMF of 80mL, anhydrous K is added₂CO₃(15.34g, 111mmol), 110 DEG C of reaction 5h, TLC detections are warming up to and are cooled to room temperature after completion of the reaction, then are poured into the ice water of 200mL, It is vigorously stirred 10min, is filtered, filter residue is washed with water 3 times (each 30mL), then is dried, and is tied again with ethyl acetate/petroleum ether Brilliant white solid (the fluoro- Isosorbide-5-Nitrae of 1- (2,4- difluorophenyl) -6,7,8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid ethyl ester, below Claim compound MH3-2) 8.61g.

Compound MH3-2 (1.53g, 4.0mmol) is added to the mixed solution of 15mLHCl (concentration 3N) and 15mL acetic acid In, 100 DEG C of reflux 3h of outer temperature are cooled to room temperature, and are filtered, and filter residue is washed with water 3 times (each 5mL), are dried in vacuo to obtain white solid (the fluoro- Isosorbide-5-Nitrae of 1- (2,4- difluorophenyl) -6,7,8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid, hereinafter referred to as compound MH3-3) 1.19g measures 250 DEG C of its mp >.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, specific spectral data is as follows, thereby determines that product is compound MH3-3.

¹H NMR (400MHz, DMSO) δ 13.97 (s, 1H), 8.83 (d, J=4.9Hz, 1H), 8.24 (s, 1H), 7.97 (s,1H),7.66 (s,1H),7.36(s,1H)；

MS(ESI,m/z):378(M+H)⁺。

Compound MH3-3 (284.2mg, 0.8mmol) is dissolved in DMSO (5.0mL), addition TEA (0.2mL, 1.44mmol) it is cooled to room temperature with compound S1-5 (259.5mg, 1.2mmol), microwave reaction 12h under conditions of 90 DEG C, It is added methylene chloride (15mL), then washes 3 times (each 15mL), it is dry, white solid is recrystallized to obtain with dehydrated alcohol 243.8mg, calculate yield be 55.26%, measure mp:218-219 DEG C.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy, carbon-13 nmr spectra are as shown in Figure 9, Figure 10, and specific spectral data is as follows, thus really Fixed output quota object is compound S5.

¹H NMR (500MHz, DMSO) δ 14.44 (s, 1H), 8.70 (s, 1H), 7.94 (d, J=11.0Hz, 2H), 7.63 (t, J=8.8Hz, 1H), 7.35 (t, J=7.5Hz, 1H), 4.74 (s, 1H), 3.95 (s, 1H), 3.76 (d, J=10.0Hz, 1H), 3.58 (s, 1H), 3.40 (s, 2H), 3.19-3.25 (m, 2H), 3.05 (d, J=10.0Hz, 2H), 1.39 (s, 9H)；

¹³C NMR(126MHz,DMSO)δ176.47,165.29,163.10,157.47,155.12,154.58,152.07, 146.05,134.63, 130.54,128.62,127.91,120.32,112.88,108.56,107.55,105.48,79.48, 57.95,53.02,50.56,49.73,28.45；

HRMS-ESI(m/z):Calcd.For C₂₆H₂₅F₄N₃NaO₆(M+Na)⁺:574.15772；Found:574.15637.

Compound S5 (113.0mg, 0.2mmol) is added to HCl (concentration 3N, 4mL)-ethyl alcohol (10mL) mixed solvent In, 100 DEG C of outer temperature is heated to reflux 10h, is cooled to room temperature, and white solid is precipitated, and filters, then wash 3 times (often with dehydrated alcohol Secondary 1mL).Obtained solid is added in 10mL dehydrated alcohol, and 90 DEG C of outer temperature is heated to reflux 30min, is cooled to room temperature, and is filtered, filter Slag is washed 3 times (each 2mL) with dehydrated alcohol again, obtains white solid 75.8mg, calculate yield be 77.69%, measure mp > 250℃。

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy is as shown in figure 27, and specific spectral data is as follows, thereby determines that product is compound S3- TM。

¹H NMR (400MHz, DMSO) δ 14.34 (s, 1H), 9.27 (br, 2H), 8.72 (s, 1H), 7.97 (t, J= 11.2Hz, 2H), 7.65 (t, J=9.6Hz, 1H), 7.36 (t, J=8.2Hz, 1H), 5.43 (s, 1H), 3.60 (s, 2H), 3.39-3.45(m,3H),3.24-3.28(m,3H), 3.07(s,1H)；

HRMS-ESI(m/z):Calcd.For C₂₁H₁₈F₄N₃O₄(M+H)⁺:452.12334；Found:452.12232.

Embodiment 6:

(S) the fluoro- 7- of -1- (2,4 difluorobenzene base) -6,8- two (4- tertbutyloxycarbonyl -3- (1- ethoxy) -1- piperazinyl) - Isosorbide-5-Nitrae-dihydro -4- Oxoquinoline -3- carboxylic acid (for formula (I) structure, hereinafter referred to as compound s 6) and its hydrochloride (hereinafter referred to as chemical combination Object S4-TM) synthesis.

Referring to the synthetic method of compound S5 in embodiment 5, by TEA be substituted for N- methylmorpholine (0.21mL, 1.92mmol), by compound S1-5 be substituted for compound S2-5 (specific synthesis process is shown in embodiment 2,368.5mg, 1.6mmol) being reacted with compound MH3-3, the methylene chloride being added after reaction is 30mL, yellow solid 203.3mg is reacted to obtain, Calculate yield be 42.18%, measure mp:210-211 DEG C.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy, carbon-13 nmr spectra are as shown in Figure 11, Figure 12, and specific spectral data is as follows, thus Determine that product is compound s 6.

¹H NMR (500MHz, DMSO) δ 14.21 (s, 1H), 8.70 (s, 1H), 7.93 (d, J=11.5Hz, 2H), 7.63 (s,1H),7.35(s, 1H),4.54(s,1H),4.01(s,1H),3.74-3.81(m,2H),3.09-3.23(m,5H),1.38 (s, 9H), 1.03 (d, J=5.5Hz, 3H)；

¹³C NMR(126MHz,DMSO)δ176.46,165.27,163.16,157.52,156.27,155.30,152.06, 146.32,134.47, 130.58,128.58,127.87,120.61,112.93,108.62,107.63,105.52,79.00, 62.87,58.06,51.34,50.59,28.49, 21.61；

HRMS-ESI(m/z):Calcd.For C₂₇H₂₇F₄N₃NaO₆(M+Na)⁺:588.17337；Found:588.17 224。

Compound s 6 (187.8mg, 3.4mmol) is added to HCl (concentration 3N, 4mL)-ethyl alcohol (1mL) in the mixed solvent, 100 DEG C of outer temperature is heated to reflux 10h, is cooled to room temperature, and white solid is precipitated, and filters, then washs 3 times (every time with dehydrated alcohol 3mL).Obtained solid is added in 15mL dehydrated alcohol, and 90 DEG C of outer temperature is heated to reflux 30min, is cooled to room temperature, and is filtered, filter residue is again Washed 3 times (each 1mL) with dehydrated alcohol, obtain white solid 139.3mg, calculate yield be 81.64%, measure mp > 250 ℃。

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy is as shown in figure 28, and specific spectral data is as follows, thereby determines that product is compound S4- TM。

¹H NMR (400MHz, DMSO) δ 14.35 (s, 1H), 9.36 (s, 1H), 9.08 (s, 1H), 8.73 (d, J= 2.8Hz, 1H), 7.96 (t, J=9.2Hz, 2H), 7.65 (s, 1H), 7.36 (s, 1H), 5.63 (s, 1H), 3.80 (s, 1H), 3.43 (d, J=14.0Hz, 3H), 3.23 (d, J=13.6 Hz, 2H), 3.04 (s, 2H), 1.12 (s, 3H)；

HRMS-ESI(m/z):Calcd.For C₂₂H₂₀F₄N₃O₄(M+H)⁺:466.13899；Found:466.13788.

Embodiment 7

(S) the fluoro- 7- of -1- (2,4 difluorobenzene base) -6,8- two (octahydro pyrrolo- [1,2-a] pyrazinyl) -1,4- dihydro -4- The synthesis of Oxoquinoline-3-carboxylic acid (for formula (VI) structure, hereinafter referred to as compound S7).

Referring to the synthetic method of compound S5 in embodiment 5, compound S1-5 is substituted for compound S3-4 (specific synthesis Process is shown in embodiment 3,151.4mg, 1.2mmol) it is reacted with compound MH3-3, the methylene chloride being added after reaction is 30mL, Yellow solid 239.8mg, calculate yield be 64.96%, measure mp:213-215 DEG C.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, as shown in figs. 13 and 14, specific spectral data is as follows, thus for gained nuclear magnetic resonance spectroscopy, carbon-13 nmr spectra Determine that product is compound S7.

¹H NMR(500MHz,CDCl₃) δ 14.41 (s, 1H), 8.47 (s, 1H), 7.94 (d, J=10.0Hz, 1H), 7.46- 7.51 (m, 1H), 7.06-7.10 (m, 2H), 3.42 (t, J=10.0Hz, 1H), 3.28-3.36 (m, 1H), 3.09 (t, J= 7.5Hz, 1H), 2.96-3.03 (m, 2H), 2.33 (t, J=10.0Hz, 1H), 2.17 (dd, J=10.0,5.0Hz, 1H), 2.09(s,1H),1.68–1.87(m,3H),1.38-1.41(m, 1H)；

¹³CNMR(126MHz,CDCl₃)δ176.77,165.93,163.35,157.46,155.13,150.39,145.34, 134.87,128.68, 128.22,127.66,119.78,112.55,108.77,108.06,105.40,62.60,55.72, 53.57,52.09,50.22,27.17,20.97；

HRMS-ESI(m/z):Calcd.For C₂₃H₂₀F₄N₃O₃(M+H)⁺:462.14408；Found:462.14303.

Embodiment 8

(S) the fluoro- 7- of -1- (2,4 difluorobenzene base) -6,8- two (7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazinyl) -1,4- The synthesis of dihydro -4- Oxoquinoline-3-carboxylic acid (for formula (VI) structure, hereinafter referred to as compound S8).

Referring to the synthetic method of compound S5 in embodiment 5, compound S1-5 is substituted for compound S4-4 (specific synthesis Process is shown in embodiment 4) it is reacted with compound MH3-3.Compound MH3-3 (142.1mg, 0.4mmol) is dissolved in DMSO In (3.0mL), TEA (0.11mL, 0.8mmol) and compound S4-4 (85.3mg, 1.2mmol) is added, under conditions of 90 DEG C Microwave reaction 12h, is cooled to room temperature, and is added methylene chloride (30mL), then wash 3 times (each 15mL), dry, with anhydrous second Alcohol recrystallizes to obtain white solid 73.3mg, and yield 38.40% measures mp:199-202 DEG C.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy, carbon-13 nmr spectra are as shown in Figure 15, Figure 16, and specific spectral data is as follows, thus Determine that product is compound S8.

¹H NMR(500MHz,DMSO)δ14.49(s,1H),8.68(s,1H),7.91-7.97(m,2H),7.61-7.65 (m, 1H), 7.34 (t, J=8.0Hz, 1H), 4.77 (d, J=5.0Hz, 1H), 4.18 (s, 1H), 3.23 (d, J=12.5Hz, 1H), 3.14 (t, J=11.0Hz, 1H), 2.88 (d, J=11.0Hz, 1H), 2.82 (t, J=11.0Hz, 1H), 2.21-2.30 (m,2H),1.96-1.99(m,1H),1.52-1.58(m,2H)；

¹³C NMR(126MHz,DMSO)δ176.49,165.33,163.06,157.41,155.01,151.97,145.60, 134.62, 130.54,128.64,127.93,119.71,112.88,108.45,107.54,105.48,67.90,63.21, 60.83,55.41,51.84,50.38, 38.93；

HRMS-ESI(m/z):Calcd.For C₂₃H₂₀F₄N₃O₄(M+H)⁺:478.13960；Foun d:478.13780.

Embodiment 9

(S) the fluoro- 7- of -1- cyclopropyl -6,8- two (4- tertbutyloxycarbonyl -3- (1- ethoxy) -1- piperazinyl) -1,4- two Hydrogen -4- Oxoquinoline-3-carboxylic acid (for formula (I) structure, hereinafter referred to as compound S9) and its hydrochloride (hereinafter referred to as compound S5- TM synthesis).

Take 2,3,4,5- tetrafluorobenzoyl ethyls (3.96g, 15mmol), be added to triethyl orthoformate (3.8mL, 22.5mmol) and in the mixed liquor of aceticanhydride (6.4mL).2.5h is reacted under conditions of 140 DEG C, vacuum distillation removes solvent and obtains Weak yellow liquid.T-BuOH (15mL) is added into the weak yellow liquid, under the protection of argon gas, ice bath is cooling, and cyclopropyl is added T-BuOH (10mL) solution of amine (0.90g, 15.8mmol) reacts 30min, reacts overnight under conditions of 45 DEG C, rotation removes Solvent is gone to obtain yellow liquid, column chromatographs to obtain yellow solid (2- (2,3,4,5- tetrafluoro benzoyl) -3- cyclopropylamino acrylic acid Ethyl ester, hereinafter referred to as compound MH1-1) 4.50g.

Compound MH1-1 (2.25g, 6.8mmol) is dissolved in the t-BuOH of 10mL, be added t-BuOH (0.76g, 6.8mmol), 5h is reacted under conditions of 90 DEG C, be cooled to room temperature after completion of the reaction, yellow solid is precipitated, filtered, filter residue is used 3 times (each 20mL) is washed, 1.31g white solid (1- cyclopropyl -6,7,8- are recrystallized to obtain with Isosorbide-5-Nitrae-dioxane after drying Three fluoro- Isosorbide-5-Nitraes-dihydro -4- Oxoquinoline-3-carboxylic acid ethyl ester, hereinafter referred to as compound MH1-2).

Compound MH1-2 (1.00g, 3.2mmol) is added to the HCl (concentration 3N) of 15mL and the solution of 15mL acetic acid In, 100 degrees Celsius of reflux 3h of outer temperature are cooled to room temperature, and are filtered, and filter residue is washed with water 3 times (each 5mL), are dried in vacuo white Solid (1- cyclopropyl -6,7, the fluoro- Isosorbide-5-Nitrae of 8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid, hereinafter referred to as compound MH1-3) 0.71g, Measure mp:223-225 DEG C.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, specific spectral data is as follows, thereby determines that product is compound MH1-3.

¹H NMR (400MHz, DMSO) δ 14.28 (s, 1H), 8.72 (s, 1H), 8.14 (d, J=8.8Hz, 1H), 4.14 (s,1H), 1.20-1.40(m,4H)。

MS(ESI,m/z):306(M+Na)⁺。

By 1- cyclopropyl -6,7, the fluoro- Isosorbide-5-Nitrae of 8- tri--dihydro -4- Oxoquinoline-3-carboxylic acid (hereinafter referred to as MH1-3,283.2mg, It 0.10mmol) is dissolved in DMSO (6.0mL), TEA (0.21mL, 1.5mmol) and compound S2-5 (specific synthesis process is added See embodiment 2,345.5mg, 1.5mmol), microwave reaction 12h under conditions of 90 DEG C is cooled to room temperature, and methylene chloride is added (30mL), then wash 3 times (each 15mL), it is dry, yellow solid 270.5mg is recrystallized to obtain with dehydrated alcohol, calculates to obtain yield It is 54.81%, measures mp:200-201 DEG C.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy, carbon-13 nmr spectra are as shown in Figure 17, Figure 18, and specific spectral data is as follows, thus Determine that product is compound S9.

¹H NMR (400MHz, DMSO) δ 14.66 (s, 1H), 8.66 (s, 1H), 7.81 (d, J=12.0Hz, 1H), 4.59 (s,1H), 4.31-4.35(m,1H),4.13-4.16(m,2H),3.82-3.89(m,2H),3.41-3.47(m,2H),3.22 (d, J=7.2Hz, 2H), 1.42 (s, 9H), 1.14-1.21 (m, 7H)；

¹³C NMR(126MHz,DMSO)δ176.17,165.70,155.35,155.38,150.70,147.61,134.4, 129.29,121.22, 107.11,79.04,62.96,57.95,56.50,51.53,50.82,40.83,28.54,21.82, 19.00,8.91；

HRMS-ESI(m/z):Calcd.For C₂₄H₃₀F₂N₃O₆(M+H)⁺:494.21027；Found:494.20854.

Referring to the synthetic method of S1-TM in embodiment 1, compound S1 is replaced with into compound S9 (49.3mg, 0.1mmol) Reacted, obtain yellow solid 32.7mg, calculate yield be 76.07%, measure 250 DEG C of mp >.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy is as shown in figure 29, thereby determines that product is compound S5-TM.

¹H NMR (400MHz, DMSO) δ 14.64 (s, 1H), 9.39 (s, 1H), 9.13 (s, 1H), 8.70 (d, J= 5.2Hz 1H), 7.88 (t, J=5.8Hz, 1H), 5.69 (s, 1H), 4.13 (s, 1H), 3.88 (s, 1H), 3.56 (s, 3H), 3.32-3.42(m,2H),3.14(s,2H),1.20(m, 7H)；

HRMS-ESI(m/z):Calcd.For C₁₉H₂₂F₂N₃O₄(M+H)⁺:394.15784；Found:394.15711.

Embodiment 10

(S) the fluoro- 7- of -1- cyclopropyl -6,8- two (4- tertbutyloxycarbonyl -3- methylol -1- piperazinyl) -1,4- dihydro -4- Oxoquinoline-3-carboxylic acid (for formula (I) structure, hereinafter referred to as compound S10) and its hydrochloride (hereinafter referred to as compound s 6-TM) Synthesis.

Referring to the synthetic method of compound S9 in embodiment 9, compound S2-5 is substituted for compound S1-5 (specific synthesis Process reacts embodiment 1,324.4mg, 1.5mmol) with compound MH1-3, obtains white solid 287.1mg, calculates Yield is 59.88%, measures mp:210-212 DEG C.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy, carbon-13 nmr spectra are as shown in Figure 19, Figure 20, and specific spectral data is as follows, thus Determine that product is compound S10.

¹H NMR (500MHz, DMSO) δ 14.71 (s, 1H), 8.68 (s, 1H), 7.83 (d, J=11.0Hz, 1H), 4.79 (t, J=5.5Hz, 1H), 4.13 (d, J=5.5Hz, 1H), 4.03 (s, 1H), 3.85 (d, J=11.5Hz, 1H), 3.68- 3.72 (m, 1H), 3.56 (d, J=12.5Hz, 1H), 3.50 (s, 1H), 3.41 (d, J=11.0Hz, 1H), 3.33 (s, 1H), 3.13-3.22(m,2H),1.43(s,9H),1.17-1.21(m,4H)；

¹³C NMR(126MHz,DMSO)δ176.13,165.7,155.07,154.69,150.58,147.23,134.6, 129.27,120.82, 107.04,106.96,79.50,58.08,53.38,50.73,50.02,40.82,28.50,19.00, 8.91；

HRMS-ESI(m/z):Calcd.For C₂₃H₂₇F₂N₃NaO₆(M+Na)⁺:502.17656；Found:502.17514.

Referring to the synthetic method of S1-TM in embodiment 1, by compound S1 replace with compound S10 (48.0mg, 0.1mmol) reacted, obtain yellow solid 31.6mg, calculate yield be 76.0%, measure 250 DEG C of mp >.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy is as shown in figure 30, and specific spectral data is as follows, thereby determines that product is compound s 6- TM。

¹H NMR (400MHz, DMSO) δ 14.64 (s, 1H), 9.36 (br, 2H), 8.70 (d, J=5.2Hz, 1H), 7.88 (d, J=7.6Hz, 1H), 5.50 (d, J=4.8Hz, 1H), 4.13 (s, 1H), 3.56-3.68 (m, 6H), 3.18 (s, 1H), 1.21 (d, J=12.4Hz, 4H)；

HRMS-ESI(m/z):Calcd.For C₁₈H₂₀F₂N₃O₄₍M+H)⁺:380.14219；Fou nd:380.14139.

Embodiment 11

(S) the fluoro- 7- of -1- cyclopropyl -6,8- two (octahydro pyrrolo- [1,2-a] pyrazinyl) -1,4- dihydro -4- oxo quinoline The synthesis of quinoline -3- carboxylic acid (for formula (VI) structure, hereinafter referred to as compound S11).

Referring to the synthetic method of compound S9 in embodiment 9, compound S2-5 is substituted for compound S3-4 and compound MH1-3 reaction.Compound MH1-3 (141.0mg, 0.5mmol) is dissolved in DMSO (3.0mL), addition TEA (0.1mL, 0.75mmol) it is cooled to room temperature with compound S1-4 (94.7mg, 0.75mmol), microwave reaction 12h under conditions of 90 DEG C, It is added methylene chloride (15 mL), then washes 3 times (each 15mL), it is dry, white solid is recrystallized to obtain with dehydrated alcohol 106.1mg, yield 54.50%, measures mp:205-207 DEG C.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, gained nuclear magnetic resonance spectroscopy, carbon-13 nmr spectra are as shown in Figure 21, Figure 22, and specific spectral data is as follows, thus Determine that product is compound S11.

¹H NMR(500MHz,CDCl₃) δ 14.59 (s, 1H), 8.75 (s, 1H), 7.86 (dd, J=12.0,2.0Hz, 1H), 3.97-4.02 (m, 1H), 3.55 (d, J=11.5Hz, 1H), 3.41-3.48 (m, 2H), 3.09-3.16 (m, 3H), 2.40- 2.45(m,1H),2.15-2.25(m,2H), 1.83-1.88(m,2H),1.74-1.78(m,1H),1.42-1.50(m,1H), 1.29-1.30(m,2H),1.16(m,2H)；

¹³C NMR(126MHz,CDCl₃)δ176.58,166.49,155.22,149.45,146.69,135.03, 128.88,120.67,108.05, 107.97,62.83,56.07,53.78,52.37,50.49,40.22,27.37,21.16, 9.22；

HRMS-ESI(m/z):Calcd.For C₂₀H₂₂F₂N₃O₃₍M+H)⁺:390.16293；Found:390.1 6168.

Embodiment 12

(S) the fluoro- 7- of -1- cyclopropyl -6,8- two (7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazinyl) -1,4- dihydro -4- oxygen For the synthesis of quinoline-3-carboxylic acid (for formula (VI) structure, hereinafter referred to as compound S12).

Referring to the synthetic method of compound S9 in embodiment 9, compound S2-5 is substituted for compound S4-4 and compound MH1-3 reaction.Compound MH1-3 (141.0mg, 0.5mmol) is dissolved in DMSO (3.0mL), addition TEA (0.1mL, 0.75mmol) and compound S2-4 (106.6mg, 0.75mmol), microwave reaction 12h under conditions of 90 DEG C are cooled to room Temperature is added methylene chloride (15mL), then washes 3 times (each 15mL), dry, recrystallizes to obtain white solid with dehydrated alcohol 69.7mg, calculate yield be 34.39%, measure mp:223-226 DEG C.

Products therefrom is divided using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer Analysis, as shown in Figure 50-52, specific spectral data is as follows for gained nuclear magnetic resonance spectroscopy, carbon-13 nmr spectra, mass spectrogram, Thereby determine that product is compound S12.

¹H NMR (500MHz, DMSO) δ 14.75 (s, 1H), 8.66 (s, 1H), 7.80 (d, J=10.0Hz, 1H), 4.81 (d, J=5.0Hz, 1H), 4.23 (d, J=5.0Hz, 1H), 4.13 (d, J=5.0Hz, 1H), 3.51 (d, J=10.0Hz, 1H), 3.33-3.40 (m, 2H), 3.27 (t, J=12.5Hz, 1H), 2.95 (t, J=10.0Hz, 2H), 2.31-2.43 (m, 2H), 2.03 (dd, J=10.0,5.0Hz, 1H), 1.57-1.65 (m, 2H), 1.17-1.21 (m, 4H)；

¹³C NMR(126MHz,DMSO)δ176.10,165.71,154.88,150.44,146.69,134.57,129.30, 120.17,107.00, 106.96,67.95,63.29,60.94,55.60,51.99,50.50,40.80,39.02,8.91；

HRMS-ESI(m/z):Calcd.For C₂₀H₂₂F₂N₃O₄(M+H)⁺:406.15784；Found:406.15634.

Embodiment 13

(R) the fluoro- 2,3- dihydro -3- methyl-1 0- of -9- (4- tertbutyloxycarbonyl -3- methylol -1- piperazinyl) -7- oxo - (3S) -7H- pyrido [1,2,3-de]-Isosorbide-5-Nitrae-benzoxazine -6- carboxylic acid (for formula (II) structure, hereinafter referred to as compound R 1) and The synthesis of its hydrochloride (hereinafter referred to as compound R 1-TM).

Side chain raw material is substituted for D-Ser, other operations entirely by reference to embodiment 1, acquisition be R configuration product. Products therefrom is analyzed using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer, gained Nuclear magnetic resonance spectroscopy is as shown in figure 31, and specific spectral data is as follows, thereby determines that product is compound R 1-TM.

¹H NMR(400MHz,DMSO)δ15.11(s,1H),9.48(br,1H),9.15(s,1H),9.00(s,1H), 7.62 (d, J=11.6Hz, 1H), 5.46 (s, 1H), 4.94 (d, J=6.4Hz, 1H), 4.61 (d, J=11.2Hz, 1H), 4.40 (d, J=11.2Hz, 1H), 3.67 (s, 2H), 3.50 (s, 3H), 3.37-3.43 (m, 2H), 3.29 (s, 1H), 3.12 (s, 1H), 1.46 (d, J=6.8Hz, 3H)；

HRMS-ESI(m/z):Calcd.ForC₁₈H₂₁FN₃O₅(M+H)⁺:378.14653；Found:378.14580

Embodiment 14

(R) the fluoro- 2,3- dihydro -3- methyl-1 0- of -9- (4- tertbutyloxycarbonyl -3- (1- ethoxy) -1- piperazinyl) -7- oxygen Generation-(3S) -7H- pyrido [1,2,3-de]-Isosorbide-5-Nitrae-benzoxazine -6- carboxylic acid (for formula (II) structure, hereinafter referred to as compound R 1) And its synthesis of hydrochloride (hereinafter referred to as compound R 2-TM).

Side chain raw material is substituted for D-Thr, other operations entirely by reference to embodiment 2, acquisition be R configuration product. Products therefrom is analyzed using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer, gained Nuclear magnetic resonance spectroscopy is as shown in figure 32, and specific spectral data is as follows, thereby determines that product is compound R 2-TM.

¹H NMR (400MHz, DMSO) δ 15.12 (s, 1H), 9.29 (s, 1H), 9.02 (m, 2H), 7.62 (dd, J= 12.0Hz J=5.6 Hz, 2H), 5.66 (s, 1H), 4.94 (s, 1H), 4.60 (d, J=7.6Hz, 1H), 4.41 (d, J= 8.8Hz, 1H), 3.86 (t, J=6.4Hz, 1H), 3.37-3.52 (m, 4H), 3.35 (d, J=10.0Hz, 1H), 3.09 (s, 2H), 1.45 (d, J=5.6Hz, 3H), 1.18 (d, J=5.6Hz, 3H)；

HRMS-ESI(m/z):Calcd.ForC₁₉H₂₃FN₃O₅(M+H)⁺:392.16217；Found:392.16152

Embodiment 15

(R) the fluoro- 7- of -1- (2,4 difluorobenzene base) -6,8- two (4- tertbutyloxycarbonyl -3- methylol -1- piperazinyl) -1,4- Dihydro -4- Oxoquinoline-3-carboxylic acid (for formula (I) structure, hereinafter referred to as compound R 5) and its hydrochloride (hereinafter referred to as compound R 3- TM synthesis).

Side chain raw material is substituted for D-Ser, other operations entirely by reference to embodiment 5, acquisition be R configuration product. Products therefrom is analyzed using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer, gained Nuclear magnetic resonance spectroscopy is as shown in figure 33, and specific spectral data is as follows, thereby determines that product is compound R 3-TM.

¹H NMR(400MHz,DMSO)δ14.36(s,1H),9.50(br,2H),9.22(br,1H),8.72(s,1H), 7.92-7.99 (m, 2H), 7.64 (t, J=10.0Hz, 1H), 7.35 (t, J=8.8Hz, 1H), 5.43 (s, 1H), 3.61 (s, 2H), 3.40-3.47 (m, 3H), 3.26 (d, J=19.6Hz, 3H), 3.06 (s, 1H)；

HRMS-ESI(m/z):Calcd.ForC₂₁H₁₈F₄N₃O₄(M+H)⁺:452.12334；Found:452.12219.

Embodiment 16

(R) the fluoro- 7- of -1- (2,4 difluorobenzene base) -6,8- two (4- tertbutyloxycarbonyl -3- (1- ethoxy) -1- piperazinyl) - Isosorbide-5-Nitrae-dihydro -4- Oxoquinoline -3- carboxylic acid (for formula (I) structure, hereinafter referred to as compound R 6) and its hydrochloride (hereinafter referred to as chemical combination Object R4-TM) synthesis.

Side chain raw material is substituted for D-Thr, other operations entirely by reference to embodiment 6, acquisition be R configuration product. Products therefrom is analyzed using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer, gained Nuclear magnetic resonance spectroscopy is as shown in figure 34, and specific spectral data is as follows, thereby determines that product is compound R 4-TM.

¹H NMR (400MHz, DMSO) δ 14.35 (s, 1H), 9.21 (br, 2H), 8.73 (d, J=2.8Hz, 1H), 7.94- 8.00 (m, 2H), 7.65 (s, 1H), 7.36 (s, 1H), 5.63 (d, J=4.4Hz, 2H), 3.78 (s, 1H), 3.41 (s, 3H), 3.23 (d, J=12.8Hz, 2H), 3.05 (s, 2H), 1.11 (s, 3H)；

HRMS-ESI(m/z):Calcd.ForC₂₂H₂₀F₄N₃O₄(M+H)⁺:466.13899；Found:466.13754.

Embodiment 17

(R) the fluoro- 7- of -1- cyclopropyl -6,8- two (4- tertbutyloxycarbonyl -3- (1- ethoxy) -1- piperazinyl) -1,4- two Hydrogen -4- Oxoquinoline-3-carboxylic acid (for formula (I) structure, hereinafter referred to as compound R 9) and its hydrochloride (hereinafter referred to as compound R 5- TM synthesis).

Side chain raw material is substituted for D-Thr, other operations entirely by reference to embodiment 9, acquisition be R configuration product. Products therefrom is analyzed using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer, gained Nuclear magnetic resonance spectroscopy is as shown in figure 35, thereby determines that product is compound R 5-TM.

¹H NMR (400MHz, DMSO) δ: 9.49 (d, J=9.6Hz, 1H), 9.17 (s, 1H), 8.68 (s, 1H), 7.86 (d, J=11.6Hz, 1H), 4.13 (d, J=4.4Hz, 1H), 3.88 (t, J=6.8Hz, 1H), 3.56-3.63 (m, 3H), 3.39-3.42 (m, 2H), 3.32 (d, J=11.6Hz, 1H), 3.12 (s, 2H), 1.20 (t, 7H)；

HRMS-ESI(m/z):Calcd.ForC₁₉H₂₂F₂N₃O₄(M+H)⁺:394.15784；Found:394.15668

Embodiment 18

(R) the fluoro- 7- of -1- cyclopropyl -6,8- two (4- tertbutyloxycarbonyl -3- methylol -1- piperazinyl) -1,4- dihydro -4- Oxoquinoline-3-carboxylic acid (for formula (I) structure, hereinafter referred to as compound R 10) and its hydrochloride (hereinafter referred to as compound R 6-TM) Synthesis.

Side chain raw material is substituted for D-Ser, other operations entirely by reference to embodiment 10, acquisition be R configuration production Object.Products therefrom is analyzed using MAT-95 type high-resolution mass spectrometer and Mercury PluS400 nuclear magnetic resonance spectrometer, Gained nuclear magnetic resonance spectroscopy is as shown in figure 36, and specific spectral data is as follows, thereby determines that product is compound s 6-TM.

¹H NMR(400MHz,DMSO)δ14.64(s,1H),9.60(s,1H),9.34(s,1H)8.69(s,1H),7.87 (d, J=11.6Hz, 1H), 5.50 (s, 1H), 4.12 (s, 1H), 3.56-3.69 (m, 5H), 3.35-3.45 (m, 3H), 3.16 (s, 1H), 1.19 (d, J=9.4Hz, 4H)；

HRMS-ESI(m/z):Calcd.ForC₁₈H₂₀F₂N₃O₄(M+H)⁺:380.14219；Found:380.14121

Antibacterial activity in vitro measurement test

1, experimental strain: R- Acinetobacter baumannii 132025355, S- Acinetobacter baumannii 130781636, R- large intestine Angstrom uncommon bacterium 132076367, S- escherichia coli 131914426, R- pseudomonas aeruginosa 131374625, S- pseudomonas aeruginosa 132083811, R- Klebsiella Pneumoniae 131636980, S- Klebsiella Pneumoniae 132099499, R- staphylococcus epidermis 131001574, R- staphylococcus epidermis GZCM, S- staphylococcus epidermis 132249588, R- staphylococcus aureus 132070039, R- staphylococcus aureus BYJ, R- staphylococcus aureus SHR, S- type staphylococcus aureus 131667100, R- enterococcus faecalis 131996907, S- enterococcus faecalis 132076451, R- Streptococcusagalactiae 131021203, S- without The B-mode secondary wound of streptococcus lactis 131636909, R- staphylococcus haemolyticus 132085699, S- staphylococcus haemolyticus 120283459, R- Cold detection of Salmonella 131667947, S- enterobacter cloacae 131127315, S- Aeromonas hydrophila 131931294, the suppurative hammer of S- Bacterium 131241647.Wherein R represents drug-resistant type；S represents responsive type.

2, MUELLER-HINTON culture medium (production of OXOID company of Britain, lot number 301651).

3, untested compound: according to embodiment 1-18 preparation Compound Compound S-1, S-2, S-4, S-5, S-6, S-7, S-8、S-9、 S-10、S-11、S-12、S1-TM、S2-TM、S3-TM、S4-TM、S5-TM、S6-TM、R1-TM、R2-TM、R3- TM, R4-TM,R5-TM,R6-TM.Untested compound is diluted with phosphate buffer solution, and concentration gradient is by many experiments 0.03125μmol/L、 0.0625μmol/L、0.125μmol/L、0.25μmol/L、0.5μmol/L、1μmol/L、2μmol/L、 4μmol/L、8μmol/L、 16μmol/L、32μmol/L、64μmol/L、128μmol/L。

4, positive control:

Reference substance 1: lefofloxacin hydrochloride；

Reference substance 2: ciprofloxacin hydrochloride.

5, test method

Antibacterial activity in vitro measurement uses the MUELLER- optimum to general bacterium of world health organisation recommendations Matrix of 1.6% agar powder as drug dilution is added in HINTON culture medium.Experimental bacteria is trained overnight in M-H broth bouillon Supporting and diluting makes inoculum concentration control be 5 × 10³-5×10⁴CFU.Drug concentration takes Consecution multiple dilution method, is added to 50-60 DEG C culture medium in prepare double dish culture dishes, 128 μm of ol/L of maximum concentration, 0.03125 μm of ol/ of minimum concentration after mixing L.Experiment bacteria suspension is inoculated with instrument with bull and is inoculated in the above-mentioned culture dish containing various compounds and various concentration.By culture dish 35-37 DEG C culture 16-18 hours, visually observe experimental result, continue culture to 42-44 hours, then observe its result.Suppression The minimum concentration of system test bacterial growth is minimum inhibitory concentration (MIC).Side chain provided by the invention is chiral monosubstituted base piperazine Antibacterial activity in vitro (MIC, μm ol/L) measurement result of the Carbostyril carboxylic acid derivatives of piperazine is shown in Tables 1 and 2.

The antibacterial activity in vitro (MIC μm of ol/L) of the part of compounds of the present invention of table 1 and positive control

--- part Gram-negative bacteria

The antibacterial activity (MIC μm of ol/L) of the part of compounds of the present invention of table 2 and positive control

--- part gram-positive bacteria

The antibacterial activity in vitro (MIC μm of ol/L) of the part of compounds of the present invention of table 3 and positive control

--- part Gram-negative bacteria

The antibacterial activity in vitro (MIC μm of ol/L) of the part of compounds of the present invention of table 4 and positive control

--- part gram-positive bacteria

By Tables 1 and 2 it can be seen that the compound in the present invention is all shown to resistance in vitro in antibacterial activity test Medicine bacterium has and the comparable antibacterial activity of reference substance.Compound in the test of Gram-negative bacteria antibacterial activity, in the present invention All show that there is preferable antibacterial activity to sensitive bacteria, wherein to the suppurative of responsive type Aeromonas hydrophila and responsive type Streptococcic antibacterial activity and reference substance Ciprofloxacin (or lefofloxacin) are suitable.In gram-positive bacteria antibacterial activity In test, individual compound such as S6-TM and R1-TM show have stronger antibacterial activity to drug-resistant type staphylococcus haemolyticus, Wherein S6-TM is 128 times of reference substance Ciprofloxacin (or lefofloxacin), and R1-TM is that reference substance Ciprofloxacin is (or left Revolve Ofloxacin) 64 times.

Compare the two class compounds that side chain is enantiomer to find, side chain be the compound of R configuration and S configuration activity how much Can be variant, active difference times is differed from 2 times to 128, but irregular, and the compound of sometimes R configuration is better than the change of S configuration Object is closed, the compound of sometimes S configuration is better than the compound of R configuration.

Above-described is only some embodiments of the present invention.For those of ordinary skill in the art, not Under the premise of being detached from the invention design, various modifications and improvements can be made, these belong to protection model of the invention It encloses.

Claims

1. a kind of quinolonecarboxylic acid compound is one of structure such as formula (II), (III) and (IV) compound represented or two Kind or more, or one or more of the nontoxic salts formed for such as formula (II), (III) and (IV) compound represented:

Wherein, the R in formula (II)₄For (S/R)-CH₂OH or (S/R)-CHOHCH₃, R₅For Boc tertbutyloxycarbonyl or H；

R in formula (III)₆For OH；

R in formula (IV)₇For OH.

2. the preparation method of one kind quinolonecarboxylic acid compound described in claim 1, it is characterised in that:

Work as R₄For (S/R)-CH₂OH, R₅When for tertbutyloxycarbonyl, formula (II) compound and its hydrochloride are made by following steps:

Lavo-ofloxacin carboxylic acid is dissolved in DMSO, TEA and 1- tertbutyloxycarbonyl -2- methylol piperazine is added, at 80~100 DEG C Under the conditions of 20~26h of microwave reaction be cooled to room temperature after the reaction was completed, methylene chloride is added, then washes 1~3 time, it is dry, use Dehydrated alcohol recrystallizes up to formula (II) compound；

Formula (II) compound is added to the in the mixed solvent of HCl- ethyl alcohol, wherein the concentration of HCl is 2~8mol/L, HCl and second The volume ratio of alcohol is 1:3~3:1, and outer temperature is heated to 100~120 DEG C of 10~12h of reflux, is cooled to room temperature, and white solid is precipitated, Filter, filter residue washs 1~3 time with dehydrated alcohol again, and obtained solid is added in dehydrated alcohol, outer temperature be heated to 90~120 DEG C with 30~50min of upper reflux, is cooled to room temperature, and filters, and filter residue washs 1~3 time with dehydrated alcohol again to get formula (II) compound Hydrochloride；

Work as R₄For (S/R)-CHOHCH₃, R₅When for tertbutyloxycarbonyl, formula (II) compound and its hydrochloride are made by following steps:

Lavo-ofloxacin carboxylic acid is dissolved in DMSO, TEA and 11- tertbutyloxycarbonyl -2- (1- ethoxy) piperazine is added, 80~ 20~26h of microwave reaction is cooled to room temperature after the reaction was completed under conditions of 100 DEG C, methylene chloride is added, then wash 1~3 time, It is dry, it is recrystallized with dehydrated alcohol up to formula (II) compound；

Work as R₆When for OH, formula (III) compound is made by following steps:

Lavo-ofloxacin carboxylic acid is dissolved in DMSO, and TEA and (S/R) -7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazine is added, 80~ 20~26h of microwave reaction is cooled to room temperature after the reaction was completed under conditions of 100 DEG C, methylene chloride is added, then wash 1~3 time, It is dry, it is recrystallized with dehydrated alcohol up to formula (III) compound；

Formula (III) compound is added to the in the mixed solvent of HCl- ethyl alcohol, wherein the concentration of HCl is 2~8mol/L, HCl and second The volume ratio of alcohol is 1:3~3:1, and outer temperature is heated to 100~120 DEG C of 10~12h of reflux, is cooled to room temperature, and white solid is precipitated, Filter, filter residue washs 1~3 time with dehydrated alcohol again, and obtained solid is added in dehydrated alcohol, outer temperature be heated to 90~120 DEG C with 30~50min of upper reflux, is cooled to room temperature, and filters, and filter residue washs 1~3 time with dehydrated alcohol again to get formula (III) compound Hydrochloride；

Work as R₇When for OH, formula (IV) compound is made by following steps:

Lavo-ofloxacin carboxylic acid is dissolved in DMSO, and TEA and (S/R) -7- hydroxyl octahydro pyrrolo- [1,2-a] pyrazine is added, 80~ 20~26h of microwave reaction is cooled to room temperature after the reaction was completed under conditions of 100 DEG C, methylene chloride is added, then wash 1~3 time, It is dry, it is recrystallized with dehydrated alcohol up to formula (IV) compound；

Formula (IV) compound is added to the in the mixed solvent of HCl- ethyl alcohol, wherein the concentration of HCl is 2~8mol/L, HCl and second The volume ratio of alcohol is 1:3~3:1, and outer temperature is heated to 100~120 DEG C of 10~12h of reflux, is cooled to room temperature, and white solid is precipitated, Filter, filter residue washs 1~3 time with dehydrated alcohol again, and obtained solid is added in dehydrated alcohol, outer temperature be heated to 90~120 DEG C with 30~50min of upper reflux, is cooled to room temperature, and filters, and filter residue washs 1~3 time with dehydrated alcohol again to get formula (IV) compound Hydrochloride.

3. claim 1 Chinese style (II), (III) or (IV) compound and its hydrochloride are preparing resisting gram-positive bacteria or anti-leather Application in Lan Shi negative bacterium drug.

4. application according to claim 3, the drug is tablet, capsule, powder needle or injection type.