克林沙星氨基衍生物及其可药用盐在制备抗结核药物中的应用转让专利
申请号 : CN201310389329.7
文献号 : CN103405435B
文献日 : 2014-12-10
发明人 : 杨大成 , 任正红 , 雷皇书 , 范莉 , 杨艳 , 韩海燕 , 邹艳冶 , 何志琴 , 赵雪晶
申请人 : 西南大学
摘要 :
本发明公开了克林沙星氨基衍生物及其可药用盐在制备抗结核药物中的应用，克林沙星氨基衍生物的结构通式中，R为-(CH2)nNR1R2、-CH(CH2CH2XCH3)NH2、2-吡咯烷基或-OR3；n为0或1；R1和R2独立地为氢、甲基、2-羟基乙基、(R)-1-乙基-2-羟基乙基、2-氨基乙基、3-(二甲基氨基)丙基、羟基、氨基、甲氨基、甲氧基或硫脲基；X为S或SO2；R3为甲基或异丁基；这些化合物对结核分枝杆菌标准敏感株、临床分离敏感株和临床分离耐药株都具有一定的抑制效果，其中部分化合物的抗结核活性强于氧氟沙星和克林沙星而略弱于莫西沙星，从而给抗结核药物提供了一个新的研究方向，有助于结核病的临床治疗。
权利要求 :
1.克林沙星氨基衍生物及其可药用盐在制备抗结核药物中的应用，所述克林沙星氨基衍生物具有以下结构通式：
1 2 3
式中，R为-(CH2)nNRR、-CH(CH2CH2XCH3)NH2、2-吡咯烷基或-OR ；
1 2
n为0或1；R 和R 独立地为氢、甲基、2-羟基乙基、(R)-1-乙基-2-羟基乙基、2-氨基乙基、3-(二甲基氨基)丙基、羟基、氨基、甲氨基、甲氧基或硫脲基；
X为S或SO2；
3
R 为甲基或异丁基。
2.根据权利要求1所述的克林沙星氨基衍生物及其可药用盐在制备抗结核药物中的
1 2
应用，其特征在于，n为0，R 和R 独立地为氢、甲基、2-羟基乙基、羟基、氨基、甲氨基或甲
1 2
氧基；n为1，R 和R 独立地为氢、甲基、2-羟基乙基、(R)-1-乙基-2-羟基乙基、2-氨基乙基、3-(二甲基氨基)丙基、羟基、氨基或硫脲基。
3.根据权利要求2所述的克林沙星氨基衍生物及其可药用盐在制备抗结核药物中的
1 2 1
应用，其特征在于，R为-(CH2)nNRR、-CH(CH2CH2XCH3)NH2、2-吡咯烷基或-OCH3；n为0，R 和
2 1 2
R 独立地为氢、甲基、羟基或氨基；n为1，R 和R 独立地为氢、甲基、2-羟基乙基、(R)-1-乙基-2-羟基乙基、2-氨基乙基、羟基、氨基或硫脲基；X为S或SO2。
4.根据权利要求3所述的克林沙星氨基衍生物及其可药用盐在制备抗结核药物中的
1 2 1 2 1 2
应用，其特征在于，R为-CH2NRR ；R 为氢，R 为硫脲基；R 为甲基或2-羟基乙基，R 为2-羟基乙基。
说明书 :
克林沙星氨基衍生物及其可药用盐在制备抗结核药物中的
应用
技术领域
[0001] 本发明属于化学领域，涉及化合物在制药领域的新用途。
背景技术
[0002] 由分支杆菌引起的结核病是一种高度传染的慢性疾病，自古以来其人群感染率一直居高不下。我国是世界上22个结核病高负担国家之一，患者人数仅次于印度居全球第二位。部分地区结核病疫情回升的原因是多方面的，有流动人口骤增、耐药结核菌的蔓延、结核菌与艾滋病病毒的双重感染等客观原因，更主要的是，普遍对结核病健康教育重视不够，结核病防治的相关知识不普及等。目前，普通结核病的一线治疗药物，如利福平(RFP)、异烟肼(INH)、吡嗪酰胺(PZA)、乙胺丁醇(EMB)和链霉素(SM)，虽然能够有效治疗普通结核病，但是伴随着治疗依从性差、周期长、有一定的毒副作用等缺点；而耐多药结核病(MDR-TB)和广泛耐药结核病(XDR TB)使结核病治疗问题越来越复杂。在过去的近50年时间里，没有一个新的预防和治疗结核病的药物推向市场。寻找或合成安全有效、治疗周期短、与抗HIV药物相容的抗分支杆菌化合物迫在眉睫。
[0003] 克林沙星(CF)曾经是第四代喹诺酮类抗菌新品种中具有“超广谱”性质的明星分子。大量体外和体内研究发现，CF不仅具有超强的抑制杀灭作用，同时显示生物利用度高、组织穿透力强、安全性和耐受性好、血清半衰期长及药代动力学性质优良等药学性质。研发单位曾将克林沙星推向三期临床，然而克林沙星固有的光毒性、心脏毒性及低血糖发生率等不良反应，妨碍了其上市申请。此外，实验过程中发现CF原药溶解性较小，水溶液中的稳定性不够，可能也是影响其成药的重要原因。
[0004] 克林沙星的衍生，最简单可行的策略就是保留克林沙星的母体结构、将其非药效基团进行结构修饰。而众多研究表明，喹诺酮的7位取代基能够很大程度上影响DNA拓扑异构酶合成及细胞通透性，并最终影响其生物活性、抗菌谱、溶解度和药代动力学。发明人对克林沙星7位吡咯环上3-氨基进行修饰衍生，合成了200多个克林沙星氨基衍生物，包括在氨基上引入五元或六元杂环、脲和胺等，从中获得了毒性降低、活性增强、溶解性提高和分子稳定性增加的抗菌活性分子(中国专利申请201010523646x,201210483417.9,201210086238.1,201310349071.8)。但这些衍生物除抗菌活性外，是否还具有新的药理活性，目前尚不清楚，有待进一步研究。
发明内容
[0005] 有鉴于此，本发明的目的在于选取部分抑菌活性较好的克林沙星氨基衍生物进行抗结核活性筛选，期望给抗结核病药物一个新方向。
[0006] 经研究，本发明提供如下技术方案：
[0007] 1.克林沙星氨基衍生物在制备抗结核病药物中的应用，所述克林沙星氨基衍生物具有以下结构通式：
[0008]
[0009] 式中，R为-(CH2)nNR1R2、-CH(CH2CH2XCH3)NH2、2-吡咯烷基或-OR3；
[0010] n为0或1；R1和R2独立地为氢、甲基、2-羟基乙基、(R)-1-乙基-2-羟基乙基、2-氨基乙基、3-(二甲基氨基)丙基、羟基、氨基、甲氨基、甲氧基或硫脲基；
[0011] X为S或SO2；
[0012] R3为甲基或异丁基。
[0013] 进一步，n为0，R1和R2独立地为氢、甲基、2-羟基乙基、羟基、氨基、甲氨基或甲氧1 2
基；n为1，R 和R 独立地为氢、甲基、2-羟基乙基、(R)-1-乙基-2-羟基乙基、2-氨基乙基、
3-(二甲基氨基)丙基、羟基、氨基或硫脲基。
基；n为1，R 和R 独立地为氢、甲基、2-羟基乙基、(R)-1-乙基-2-羟基乙基、2-氨基乙基、
3-(二甲基氨基)丙基、羟基、氨基或硫脲基。
[0014] 进一步，R为-(CH2)nNR1R2、-CH(CH2CH2XCH3)NH2、2-吡咯烷基或-OCH3；n为0，R1和2 1 2
R 独立地为氢、甲基、羟基或氨基；n为1，R 和R 独立地为氢、甲基、2-羟基乙基、(R)-1-乙基-2-羟基乙基、2-氨基乙基、羟基、氨基或硫脲基；X为S或SO2。
R 独立地为氢、甲基、羟基或氨基；n为1，R 和R 独立地为氢、甲基、2-羟基乙基、(R)-1-乙基-2-羟基乙基、2-氨基乙基、羟基、氨基或硫脲基；X为S或SO2。
[0015] 进一步，R为-CH2NR1R2；R1为氢，R2为硫脲基；R1为甲基或2-羟基乙基，R2为2-羟基乙基。
[0016] 本发明的有益效果在于：本发明从部分抑菌活性较好的克林沙星氨基衍生物中筛选出了具有抗结核活性的化合物，这些化合物对结核分枝杆菌标准敏感株、临床分离敏感株和临床分离耐药株都具有一定的抑制效果，其中部分化合物的抗结核活性强于氧氟沙星和克林沙星而略弱于莫西沙星，从而给抗结核药物提供了一个新的研究方向，有助于结核病的临床治疗。
具体实施方式
[0017] 为了使本发明的目的、技术方案和有益效果更加清楚，下面对本发明的优选实施例进行详细的描述。
[0018] 实施例1、化合物TM1的合成
[0019]
[0020] 向反应瓶中加入克林沙星(CF)、K2CO3及适量二氯甲烷(DCM)，在冰浴下滴加氯乙酰氯的DCM溶液，CF、K2CO3和氯乙酰氯的投料摩尔比为10:25:15～25，滴毕，冰浴下搅拌反应，TLC(薄层色谱法)监测反应进程。反应完毕，得到黄绿色浑浊液，抽滤，滤饼用DCM洗涤，洗液与滤液合并，旋蒸至干，得中间体IM1。
[0021] 向反应瓶中加入胺组分、K2CO3及催化量的KI和适量溶剂，室温搅拌30分钟后加入中间体IM1，IM1、K2CO3与胺组分的投料摩尔比为1:3:2，30-45℃控温反应，TLC监测反应进程。反应完毕，根据具体情况分别按以下几种方式进行后处理：(1)若产物溶于有机溶剂，加水，用2N HCl调pH至6-7，用DCM萃取，收集有机相，旋蒸得粗品，用二氯甲烷-石油醚(DCM-PE)重结晶；(2)若产物溶于水，调pH至中性，静置分层，收集水相，继续调pH至4～5，冷藏析晶，抽滤，滤饼用90%乙醇重结晶；(3)若反应体系有固体产生，抽滤，滤饼加水，用
2N HCl调pH至4～5，冷藏析晶；(4)若产物既溶于有机溶剂，又溶于水，先旋蒸除去有机相，再用2N HCl调pH至4～5，冷藏析晶，即得化合物TM1。合成结果见表1。
2N HCl调pH至4～5，冷藏析晶；(4)若产物既溶于有机溶剂，又溶于水，先旋蒸除去有机相，再用2N HCl调pH至4～5，冷藏析晶，即得化合物TM1。合成结果见表1。
[0022] 表1化合物TM1合成结果
[0023]
[0024]
[0025] TM1-17-(3-(2-( 羟 胺 基 ) 乙 酰 胺 基 )-1- 吡 咯 烷 基 )-1- 环 丙1
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：浅黄色固体,m.p.223.2～224.1℃;H NMR(600MHz,DMSO-d6)δ:14.50(s,1H,H-11),8.82(s,1H,H-2),7.91(d,1H,J=12.0Hz,H-5),4.41-4.36(m,1H,H-12),3.80-3.58(m,5H,H-16、H-20and H-17),3.38-3.33(m,4H,H-14and
13
H-15),1.21-1.17(m,2H,H-13),1.01-0.98(m,2H,H-13); C NMR(151MHz,DMSO-d6)δ:176.07,166.73,165.06,(156.30,154.64),152.79,(143.59,143.50),137.95,(122.83,122.78),(119.48,119.45),(110.63,110.47),107.63,62.64,51.05,50.66,45.97,41.94,41.56,10.84(2×C); (c=1.5mg/mL,DMF).
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：浅黄色固体,m.p.223.2～224.1℃;H NMR(600MHz,DMSO-d6)δ:14.50(s,1H,H-11),8.82(s,1H,H-2),7.91(d,1H,J=12.0Hz,H-5),4.41-4.36(m,1H,H-12),3.80-3.58(m,5H,H-16、H-20and H-17),3.38-3.33(m,4H,H-14and
13
H-15),1.21-1.17(m,2H,H-13),1.01-0.98(m,2H,H-13); C NMR(151MHz,DMSO-d6)δ:176.07,166.73,165.06,(156.30,154.64),152.79,(143.59,143.50),137.95,(122.83,122.78),(119.48,119.45),(110.63,110.47),107.63,62.64,51.05,50.66,45.97,41.94,41.56,10.84(2×C); (c=1.5mg/mL,DMF).
[0026] TM1-27-(3-(2-((R)-1-羟基丁醇-2-胺基)乙酰胺基)-1-吡咯烷基)-1-环丙基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：浅黄色固体,m.p.233.7～1
235.5 ℃ ;H NMR(300MHz,DMSO-d6)δ:14.52(s,1H,H-11),8.86(s,1H,H-2),8.68(s,1H,CONH),7.99(d,1H,J=11.4Hz,H-5),5.41(s,1H,H-24),4.43-4.39(m,1H,H-12),4.17(s,2H,H-20),3.73-3.60(m,6H,H-16、H-17、H-14and H-23),3.39-3.34(m,3H,H-14and H-15),3.08-3.06(m,1H,H-22),1.69-1.53(m,2H,CH3CH2),1.20-1.18(m,2H,H-13),1.01-0.91(
13
m,5H,H-13and CH3CH2); C NMR(75MHz,DMSO-d6)δ:176.17,165.14,164.35,(157.30,15
3.97),152.92,(143.43,143.24),137.92,(123.20,123.09),119.88,(110.81,110.50),
107.68,60.60,58.22,50.74,50.38,,45.11,44.70,42.37,41.65,20.28,10.92,9.99;HR +
MS:C23H28ClFN4O5[M+H] 计算值为495.1805,实测值为495.1809; (c=1.5mg/mL,H2O).
235.5 ℃ ;H NMR(300MHz,DMSO-d6)δ:14.52(s,1H,H-11),8.86(s,1H,H-2),8.68(s,1H,CONH),7.99(d,1H,J=11.4Hz,H-5),5.41(s,1H,H-24),4.43-4.39(m,1H,H-12),4.17(s,2H,H-20),3.73-3.60(m,6H,H-16、H-17、H-14and H-23),3.39-3.34(m,3H,H-14and H-15),3.08-3.06(m,1H,H-22),1.69-1.53(m,2H,CH3CH2),1.20-1.18(m,2H,H-13),1.01-0.91(
13
m,5H,H-13and CH3CH2); C NMR(75MHz,DMSO-d6)δ:176.17,165.14,164.35,(157.30,15
3.97),152.92,(143.43,143.24),137.92,(123.20,123.09),119.88,(110.81,110.50),
107.68,60.60,58.22,50.74,50.38,,45.11,44.70,42.37,41.65,20.28,10.92,9.99;HR +
MS:C23H28ClFN4O5[M+H] 计算值为495.1805,实测值为495.1809; (c=1.5mg/mL,H2O).
[0027] TM1-37-(3-(2-(羟 乙 基 胺 基) 乙 酰 胺 基 )-1-吡 咯 烷 基 )-1-环 丙1
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：淡黄色固体,m.p.239.5-240.2℃;H NMR(300MHz,DMSO-d6)δ:14.50(s,1H,H-11),8.93(s,1H,H-18),8.84(s,1H,H-2),7.97(d,
2H,J=12Hz,H-5),5.26(s,1H,-OH),4.33-4.26(m,1H,H-12),4.15(s,2H,H-20),3.71-3.68(m,3H,H-16and H-17),3.58-3.55(m,2H,H-23),3.38-3.31(m,4H,H-14and H-15),3.01-2.
99(m,2H,H-22),2.87-2.78(m,2H,H-21),1.21-1.18(m,2H,H-13),1.03-0.99(m,2H,H-13)
13
; C NMR(75MHz,DMSO-d6)δ:176.52,165.46,164.43,(157.42,154.27),153.27,143.61,1
38.27,123.56,120.34,(111.15,110.85),108.07,50.97,49.16,48.31,45.54,42.56,42.+
02,30.41,29.37,11.56,11.37;HR MS:C21H24ClFN4O5[M+Na] 计算值为489.1311,实测值为
489.1315; (c=1.0mg/mL,CHCl3).
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：淡黄色固体,m.p.239.5-240.2℃;H NMR(300MHz,DMSO-d6)δ:14.50(s,1H,H-11),8.93(s,1H,H-18),8.84(s,1H,H-2),7.97(d,
2H,J=12Hz,H-5),5.26(s,1H,-OH),4.33-4.26(m,1H,H-12),4.15(s,2H,H-20),3.71-3.68(m,3H,H-16and H-17),3.58-3.55(m,2H,H-23),3.38-3.31(m,4H,H-14and H-15),3.01-2.
99(m,2H,H-22),2.87-2.78(m,2H,H-21),1.21-1.18(m,2H,H-13),1.03-0.99(m,2H,H-13)
13
; C NMR(75MHz,DMSO-d6)δ:176.52,165.46,164.43,(157.42,154.27),153.27,143.61,1
38.27,123.56,120.34,(111.15,110.85),108.07,50.97,49.16,48.31,45.54,42.56,42.+
02,30.41,29.37,11.56,11.37;HR MS:C21H24ClFN4O5[M+Na] 计算值为489.1311,实测值为
489.1315; (c=1.0mg/mL,CHCl3).
[0028] TM1-47-(3-(2-(N- 甲 基 - 羟 乙 基 胺 基 ) 乙 酰 胺 基 )-1- 吡 咯 烷基)-1-环丙基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：淡黄色固体,m.1
p.130.7-131.4 ℃ ;H NMR(300MHz,DMSO-d6)δ:14.55(s,1H,H-11),9.59(s,1H,H-18),
8.88(s,1H,H-2),8.00(d,2H,J=12.0Hz,H-5),5.40(s,1H,-OH),4.50-4.39(m,3H,H-12and H-20),3.88-3.66(m,4H,H-16、H-17and H-14),3.56-3.53(m,2H,H-22),3.42-3.39(m,3H,H-14and H-15),3.25-3.18(m,2H,H-21),2.92(s,3H,CH3),1.22-1.19(m,2H,H-13
13
),1.02-0.93(m,2H,H-13); C NMR(75MHz,DMSO)δ:176.52,165.46,164.43,(157.43,1
54.58),153.27,(143.61,143.45),138.27,123.56,120.34,(111.15,110.85),108.07,
62.86,61.19,58.34,50.97,49.72,48.31,45.54,42.56,42.02,30.41,11.53,11.34;HR +
MS:C22H26ClFN4O5[M+Na] 计算值为503.1468,实测值为503.1465; (c=1.0mg/mL,CHCl3).
p.130.7-131.4 ℃ ;H NMR(300MHz,DMSO-d6)δ:14.55(s,1H,H-11),9.59(s,1H,H-18),
8.88(s,1H,H-2),8.00(d,2H,J=12.0Hz,H-5),5.40(s,1H,-OH),4.50-4.39(m,3H,H-12and H-20),3.88-3.66(m,4H,H-16、H-17and H-14),3.56-3.53(m,2H,H-22),3.42-3.39(m,3H,H-14and H-15),3.25-3.18(m,2H,H-21),2.92(s,3H,CH3),1.22-1.19(m,2H,H-13
13
),1.02-0.93(m,2H,H-13); C NMR(75MHz,DMSO)δ:176.52,165.46,164.43,(157.43,1
54.58),153.27,(143.61,143.45),138.27,123.56,120.34,(111.15,110.85),108.07,
62.86,61.19,58.34,50.97,49.72,48.31,45.54,42.56,42.02,30.41,11.53,11.34;HR +
MS:C22H26ClFN4O5[M+Na] 计算值为503.1468,实测值为503.1465; (c=1.0mg/mL,CHCl3).
[0029] TM1-57-(3-(2-(二 羟乙 基 胺 基)乙 酰 胺 基)-1-吡 咯 烷基 )-1-环 丙1
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：淡黄色固体,m.p.242.3-242.8℃;H NMR(300MHz,DMSO-d6)δ:14.48(s,1H,H-11),9.64(s,1H,H-18),8.84(s,1H,H-2),7.9
8(d,2H,J=12Hz,H-5),5.21(s,2H,OH),4.56-4.40(m,1H,H-12),4.16(s,2H,H-20),3.
82-3.74(m,3H,H-16and H-17),3.64(t,4H,J=6.0Hz,H-22),3.59-3.48(m,6H,H-14an
13
d H-23),3.00-2.98(m,2H,H-15),1.22-1.19(m,2H,H-13),1.04-1.00(m,2H,H-13);
C NMR(75MHz,DMSO-d6)δ:176.04,168.24,164.98,(156.32,154.66),152.82,(143.2
9,143.20),137.82,(123.02,122.97),(119.71,119.68),(110.62,110.46),107.58,5
5.12(2×C),54.69(2×C),50.65,50.45,49.79,45.48,41.77,41.46,10.73(2×C);HR +
MS:C23H28ClFN4O6[M+Na] 计算值为533.1574,实测值为533.1576; (c=1.0mg/mL,CHCl3).
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：淡黄色固体,m.p.242.3-242.8℃;H NMR(300MHz,DMSO-d6)δ:14.48(s,1H,H-11),9.64(s,1H,H-18),8.84(s,1H,H-2),7.9
8(d,2H,J=12Hz,H-5),5.21(s,2H,OH),4.56-4.40(m,1H,H-12),4.16(s,2H,H-20),3.
82-3.74(m,3H,H-16and H-17),3.64(t,4H,J=6.0Hz,H-22),3.59-3.48(m,6H,H-14an
13
d H-23),3.00-2.98(m,2H,H-15),1.22-1.19(m,2H,H-13),1.04-1.00(m,2H,H-13);
C NMR(75MHz,DMSO-d6)δ:176.04,168.24,164.98,(156.32,154.66),152.82,(143.2
9,143.20),137.82,(123.02,122.97),(119.71,119.68),(110.62,110.46),107.58,5
5.12(2×C),54.69(2×C),50.65,50.45,49.79,45.48,41.77,41.46,10.73(2×C);HR +
MS:C23H28ClFN4O6[M+Na] 计算值为533.1574,实测值为533.1576; (c=1.0mg/mL,CHCl3).
[0030] TM1-67-(3-(2-(2-氨 乙 基 胺 基)乙 酰 胺 基)-1-吡 咯 烷 基)-1-环 丙基-6-氟-8-氯-1,4-二氢 -4-氧代-3-喹啉 羧酸：浅黄色固 体,m.p.241.2～1
242.3℃;H NMR(300MHz,DMSO-d6)δ:14.50(s,1H,H-11),8.85(s,1H,H-2),7.97(d,1H,J=
11.7Hz,H-5),4.43-4.39(m,1H,H-12),4.27(s,2H,H-20),3.72-3.58(m,4H,H-16、H-17and H-14),3.40-3.27(m,7H,H-14、H-22、H-23and H-15),1.22-1.19(m,2H,H-13),1.02-0.99(m
13
,2H,H-13); C NMR(75MHz,DMSO-d6)δ:176.09,165.08,163.93,(157.26,153.94),152.86,(143.42,143.22),137.89,(123.13,123.03),(119.93,119.86),(110.75,110.44),107.6
4,50.89,50.60,47.23,45.12,44.16,42.21,41.64,35.00,10.92(2×C);HRMS:C21H25ClFN5O+
4[M+H] 计算值为466.1652,实测值为466.1658; (c=1.5mg/mL,H2O).
242.3℃;H NMR(300MHz,DMSO-d6)δ:14.50(s,1H,H-11),8.85(s,1H,H-2),7.97(d,1H,J=
11.7Hz,H-5),4.43-4.39(m,1H,H-12),4.27(s,2H,H-20),3.72-3.58(m,4H,H-16、H-17and H-14),3.40-3.27(m,7H,H-14、H-22、H-23and H-15),1.22-1.19(m,2H,H-13),1.02-0.99(m
13
,2H,H-13); C NMR(75MHz,DMSO-d6)δ:176.09,165.08,163.93,(157.26,153.94),152.86,(143.42,143.22),137.89,(123.13,123.03),(119.93,119.86),(110.75,110.44),107.6
4,50.89,50.60,47.23,45.12,44.16,42.21,41.64,35.00,10.92(2×C);HRMS:C21H25ClFN5O+
4[M+H] 计算值为466.1652,实测值为466.1658; (c=1.5mg/mL,H2O).
[0031] TM1-77-(3-(2-((3-二甲氨基丙基)胺基)乙酰胺基)-1-吡咯烷基)-1-环丙基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：浅黄色固体,m.p.181.8～183.6℃;1H NMR(300MHz,DMSO-d6)δ:14.47(s,1H,H-11),8.84(s,1H,H-2),8.64(s,1H,H-18),7.94(d,1H,J=11.7Hz,H-5),4.44-4.39(m,1H,H-12),4.22(s,2H,H-20),3.71-3.61(m,4H,H-16、H-17and H-14),3.42-3.33(m,3H,H-14andH-15),3.21-3.07(m,4H,H-22and H-24),2.89(d,1H,J=7.5Hz,H-21),2.76(s,6H,CH3),2.22-2.18(m,2H,H-23),1.24-1.19(m,2H,H-13),1.04-1.01(m,2H,H-13);13C NMR(75MHz,DMSO-d6)δ:176.03,165.06,164.05,(157.20,15
3.88),152.80,(143.41,143.22),137.86,(123.07,122.96),119.80,(110.71,110.40),1
07.61,53.37,51.45,50.58,46.86,45.15,44.18,41.91(2×C),41.65,36.11,20.43,10.9+
3(2×C);HR MS:C24H31ClFN5O4[M+H] 计算值为508.2121,实测值为508.2129;
(c=1.5mg/mL,H2O).
3.88),152.80,(143.41,143.22),137.86,(123.07,122.96),119.80,(110.71,110.40),1
07.61,53.37,51.45,50.58,46.86,45.15,44.18,41.91(2×C),41.65,36.11,20.43,10.9+
3(2×C);HR MS:C24H31ClFN5O4[M+H] 计算值为508.2121,实测值为508.2129;
(c=1.5mg/mL,H2O).
[0032] TM1-87-(3-(2-(肼基)乙酰胺基)-1-吡咯烷基)-1-环丙基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：黄色固体,m.p.164.6～166.2℃;1H NMR(600MHz,DMSO-d6)δ:14.51(s,1H,H-11),8.84(s,1H,H-2),7.94(d,1H,J=12.0Hz,H-5),4.43-4.39(m,1H,H-12),3.96(s,2H,H-20),3.73-3.64(m,3H,H-16and H-17),3.58-3.56(m,2H,H-22),3.38-3.33(m,4H,H-14and H-15),1.21-1.19(m,2H,H-13),1.02-0.97(m,2H,H-13);13C NMR(151MHz,DMSO-d6)δ:176.13,165.12(2×C),(156.84,154.34),152.92,(143.43,143.
29),137.93,(123.13,123.05),(119.87,119.82),(110.76,110.53),107.67,50.78,50.62,49.45,45.01,42.03,41.64,10.91(2×C);HR MS:C28H35ClFN5O6[M+H]+计算值为438.1339,实测值为438.1340; (c=1.5mg/mL,DMF).
29),137.93,(123.13,123.05),(119.87,119.82),(110.76,110.53),107.67,50.78,50.62,49.45,45.01,42.03,41.64,10.91(2×C);HR MS:C28H35ClFN5O6[M+H]+计算值为438.1339,实测值为438.1340; (c=1.5mg/mL,DMF).
[0033] TM1-97-(3-(2-(1-甲 基 肼 基 ) 乙 酰 胺 基 )-1- 吡 咯 烷 基)-1- 环 丙1
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：黄色固体,m.p.157.1～158.7℃;H NMR(300MHz,DMSO-d6)δ:14.52(s,1H,H-11),8.86(s,1H,H-2),7.97(d,1H,J=11.7Hz,H-5),4.43-4.39(m,1H,H-12),3.94(s,2H,H-20),3.68-3.61(m,4H,H-16、H-17and H-14),3.38-3.33(m,3H,H-14and H-15),3.47(s,3H,CH3),1.22-1.19(m,2H,H-13),1.02-0.
13
99(m,2H,H-13); C NMR(75MHz,DMSO-d6)δ:176.09,166.40,165.08,(157.23,153.91),15
2.84,(143.45,143.26),137.91,(123.08,122.97),119.76,(110.74,110.44),107.65,56.+
97,50.84,50.57,45.51,43.35,41.90,41.63,10.90(2×C);HR MS:C20H23ClFN5O4[M+H] 计算值为452.1495,实测值为452.1504; (c=1.0mg/mL,DMF).
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：黄色固体,m.p.157.1～158.7℃;H NMR(300MHz,DMSO-d6)δ:14.52(s,1H,H-11),8.86(s,1H,H-2),7.97(d,1H,J=11.7Hz,H-5),4.43-4.39(m,1H,H-12),3.94(s,2H,H-20),3.68-3.61(m,4H,H-16、H-17and H-14),3.38-3.33(m,3H,H-14and H-15),3.47(s,3H,CH3),1.22-1.19(m,2H,H-13),1.02-0.
13
99(m,2H,H-13); C NMR(75MHz,DMSO-d6)δ:176.09,166.40,165.08,(157.23,153.91),15
2.84,(143.45,143.26),137.91,(123.08,122.97),119.76,(110.74,110.44),107.65,56.+
97,50.84,50.57,45.51,43.35,41.90,41.63,10.90(2×C);HR MS:C20H23ClFN5O4[M+H] 计算值为452.1495,实测值为452.1504; (c=1.0mg/mL,DMF).
[0034] TM1-107-(3-(2-(硫代 脲基胺 基)乙酰胺 基)-1-吡咯 烷基)-1-环 丙1
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：土黄色固体,m.p.287.5～289.2℃;H NMR(300MHz,DMSO-d6)δ:8.86(s,1H,H-2),8.01(d,1H,J=11.7Hz,H-5),6.01(s,2H,H-20),
4.43-4.38(m,1H,H-12),3.56-3.50(m,5H,H-20、H-16and H-17),3.27-3.20(m,4H,H-14and H-17),1.20-1.18(m,2H,H-13),1.03-1.00(m,2H,H-13); (c=1.5mg/mL,H2O).[0035] 实施例2、化合物TM2的合成
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：土黄色固体,m.p.287.5～289.2℃;H NMR(300MHz,DMSO-d6)δ:8.86(s,1H,H-2),8.01(d,1H,J=11.7Hz,H-5),6.01(s,2H,H-20),
4.43-4.38(m,1H,H-12),3.56-3.50(m,5H,H-20、H-16and H-17),3.27-3.20(m,4H,H-14and H-17),1.20-1.18(m,2H,H-13),1.03-1.00(m,2H,H-13); (c=1.5mg/mL,H2O).[0035] 实施例2、化合物TM2的合成
[0036]
[0037] 向反应瓶中加入CF和适量氯仿，搅拌均匀后，缓慢滴加二(三氯甲基)碳酸酯(BTC)的氯仿溶液，滴加完毕，冰浴下剧烈搅拌反应7小时，再缓慢滴加三乙胺(TEA)，CF、BTC和TEA的投料摩尔比为17:6:10，滴加完毕，室温下反应，TLC监测反应进程。反应完毕，抽滤，滤饼用DCM洗涤，洗液与滤液合并，加水，用2N HCl调pH至4～5，再用DCM萃取2次，收集有机相，用饱和食盐水洗涤2次，无水硫酸钠干燥，过滤，滤液旋蒸浓缩，快速柱层析法纯化，得中间体IM2。
[0038] 向反应瓶中加入氯仿、胺组分和K2CO3，室温搅拌0.5小时后，加入中间体IM2，胺组分、K2CO3和IM2的投料摩尔比为1.5:2:1，控温反应，TLC监测反应进程。反应完毕，根据具体情况分别按以下几种方式进行后处理：(1)对于水溶性较差的产物，抽滤，滤饼用DCM洗涤后，加水，调pH至弱酸性，用DCM萃取，收集有机相，用DCM-PE(石油醚)重结晶；(2)对于水溶性较好的产物，调pH至弱酸性，加入固体食盐至饱和，再用DCM萃取，收集有机相，用DCM-PE(石油醚)重结晶；或者，调pH至弱酸性，加入固体食盐至饱和，置冰箱中冷藏析晶，次日抽滤，即得化合物TM2。合成结果见表2。
[0039] 表2化合物TM2合成结果
[0040]
[0041] TM2-17-(3-脲-1-吡咯烷基)-1-环丙基-6-氟-8-氯-1,4-二氢-4-氧1
代-3-喹啉羧酸：m.p.253.7-254.9 ℃ H NMR(600MHz,DMSO-d6)δ:14.34(s,1H,H-11),
8.83(s,1H,H-2),7.92(d,1H,J=12Hz,H-5),6.12(s,2H,H-20),5.76(m,1H,H-18),4.41-4.38(m,1H,H-12),3.48-3.43(m,3H,H-14、H-15and H-17),3.30-3.27(m,4H,H-14、H-15、
13
H-16and H-17),1.22-1.19(m,2H,H-13),1.01-0.99(m,2H,H-13); C NMR(151MHz,DMSO-d6)δ:(176.55,176.53),(165.51,158.56),156.83,155.16,155.22,(144.12,144.02),1
38.38,(123.28,123.23),(120.03,120.00),(111.06,110.91),108.06,51.21,51.18,44.63(2×C),42.02,11.27(2×C);HR MS C18H18ClFN4NaO4计算值为431.0893,实测值为
431.0896; (c=1mg/mL,DMF).
代-3-喹啉羧酸：m.p.253.7-254.9 ℃ H NMR(600MHz,DMSO-d6)δ:14.34(s,1H,H-11),
8.83(s,1H,H-2),7.92(d,1H,J=12Hz,H-5),6.12(s,2H,H-20),5.76(m,1H,H-18),4.41-4.38(m,1H,H-12),3.48-3.43(m,3H,H-14、H-15and H-17),3.30-3.27(m,4H,H-14、H-15、
13
H-16and H-17),1.22-1.19(m,2H,H-13),1.01-0.99(m,2H,H-13); C NMR(151MHz,DMSO-d6)δ:(176.55,176.53),(165.51,158.56),156.83,155.16,155.22,(144.12,144.02),1
38.38,(123.28,123.23),(120.03,120.00),(111.06,110.91),108.06,51.21,51.18,44.63(2×C),42.02,11.27(2×C);HR MS C18H18ClFN4NaO4计算值为431.0893,实测值为
431.0896; (c=1mg/mL,DMF).
[0042] TM2-27-(3-(3-甲基脲)-1-吡咯烷基)-1-环丙基-6-氟-8-氯-1,4-二氢-4-氧1
代-3-喹啉羧酸：m.p.212.9-215.8℃;H NMR(300MHz,CDCl3)δ:14.26(s,1H,H-11),8.
93(s,1H,H-2),8.05(d,1H,J=9Hz,H-5),5.30(s,1H,H-18),4.61-4.59(m,1H,H-20),4.40-
4.33(m,1H,H-12),3.93-3.85(m,4H,H-14、H-15、H-16and H-17),3.43-3.42(m,3H,H-14、
13
H-15and H-17),3.37-3.28(d,3H,J=9Hz,H-21),1.36-0.96(m,4H,H-13); C NMR(75MHz,CDCl3)δ:176.63,165.82,158.05,154.72,148.58,(143.52,143.26),137.7
2,(124.63,124.48),(120.74,120.59),(112.12,111.81),108.53,50.93,50.44,49.11,46.62,41.35,35.76,11.43(2×C);HR MS:C19H21ClFN4O4计算值为423.1230,实测值为
423.1236; (c=1mg/mL,DCM).
代-3-喹啉羧酸：m.p.212.9-215.8℃;H NMR(300MHz,CDCl3)δ:14.26(s,1H,H-11),8.
93(s,1H,H-2),8.05(d,1H,J=9Hz,H-5),5.30(s,1H,H-18),4.61-4.59(m,1H,H-20),4.40-
4.33(m,1H,H-12),3.93-3.85(m,4H,H-14、H-15、H-16and H-17),3.43-3.42(m,3H,H-14、
13
H-15and H-17),3.37-3.28(d,3H,J=9Hz,H-21),1.36-0.96(m,4H,H-13); C NMR(75MHz,CDCl3)δ:176.63,165.82,158.05,154.72,148.58,(143.52,143.26),137.7
2,(124.63,124.48),(120.74,120.59),(112.12,111.81),108.53,50.93,50.44,49.11,46.62,41.35,35.76,11.43(2×C);HR MS:C19H21ClFN4O4计算值为423.1230,实测值为
423.1236; (c=1mg/mL,DCM).
[0043] TM2-37-(3-肼基甲酰胺-1-吡咯烷基)-1-环丙基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸：m.p.221.2.1-223.7℃;1H NMR(300MHz,DMSO-d6)δ:14.52(s,1H,H-11),8.83(s,1H,H-2),7.94(d,1H,J=12Hz,H-5),5.75(m,1H,H-18),4.42-4.34(m,1H,H-20),3.4
8-3.43(m,4H,H-12、H-14、H-15and H-17),3.35-3.30(m,4H,H-14、H-15、H-16and H-17),1.
18-0.98(m,4H,H-13);13CNMR(75MHz,DMSO-d6)δ:(176.65,176.62),165.84,157.88,154.5
3,151.97,(144.21,144.01),137.87,(123.78,123.67),(120.05,119.99),(111.80,111.4
8),108.43,51.18,51.04,44.48(2×C),41.98,11.25(2×C);HR MS:C18H20ClFN5O4计算值为
424.1182,实测值为424.1185; (c=1mg/mL,DMF).
8-3.43(m,4H,H-12、H-14、H-15and H-17),3.35-3.30(m,4H,H-14、H-15、H-16and H-17),1.
18-0.98(m,4H,H-13);13CNMR(75MHz,DMSO-d6)δ:(176.65,176.62),165.84,157.88,154.5
3,151.97,(144.21,144.01),137.87,(123.78,123.67),(120.05,119.99),(111.80,111.4
8),108.43,51.18,51.04,44.48(2×C),41.98,11.25(2×C);HR MS:C18H20ClFN5O4计算值为
424.1182,实测值为424.1185; (c=1mg/mL,DMF).
[0044] TM2-47-(3-(1-甲基肼基甲酰胺-1-吡咯烷基)-1-环丙基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸盐酸盐：m.p.266.7-269.3℃,1H NMR(600MHz,DMSO-d6)δ:14.34(s,1H,H-11),10.15(s,3H,H-22and H-23),8.86(s,1H,H-2),7.98(d,1H,J=12Hz,H-5),5.76(s,1H,H-18),4.43-4.40(m,1H,H-12),3.39-3.36(m,3H,H-14、H-15and H-17),3.37-3.34(m,4H,H-14、H-15、H-16and H-17),3.07(s,3H,H-21),1.22-1.19(m,2H,H-13),1.02-0.99(m,2H,H-13);13C NMR(151MHz,DMSO-d6)δ:176.60,165.51,161.67,(1
56.91,155.23),153.39,(143.72,143.62),138.34,(123.72,123.67),(120.44,120.41),(111.20,111.04),108.14,50.92,50.89,46.32(2×C),42.02,41.36,11.29(2×C);HR MS C19H22ClFN5O4计算值为438.1339,实测值为438.1336; (c=1mg/mL,DMF).
56.91,155.23),153.39,(143.72,143.62),138.34,(123.72,123.67),(120.44,120.41),(111.20,111.04),108.14,50.92,50.89,46.32(2×C),42.02,41.36,11.29(2×C);HR MS C19H22ClFN5O4计算值为438.1339,实测值为438.1336; (c=1mg/mL,DMF).
[0045] TM2-57-(3-(3-羟基脲)-1-吡咯烷基)-1-环丙基-6-氟-8-氯-1,4-二1
氢-4-氧 代 -3-喹 啉 羧 酸：m.p.219.3-222.0 ℃ ,H NMR(600MHz,DMSO-d6)δ:14.34(s,1H,H-11),9.20(s,1H,H-21),8.83(s,1H,H-2),8.13(s,1H,H-20),7.92(d,1H,J=12Hz,H-5),5.76(s,1H,H-18),4.40-4.38(m,1H,H-12),3.48-3.44(m,3H,H-14、H-15and H-17),3.30-3.27(m,4H,H-14、H-15、H-16and H-17),1.22-1.19(m,2H,H-13),1.02-0.99
13
(m,2H,H-13); C NMR(151MHz,DMSO-d6)δ:176.53,165.52,160.13,(156.84,155.18),15
3.25,(143.99,143.90),138.35,(123.42,123.37),(120.17,120.14),(111.09,110.93),108.12,51.07,51.04,44.40(2×C),42.00,11.27(2×C);HR MS C18H19ClFN4O5计算值为
425.1022,实测值为425.1027; (c=1mg/mL,DMF).
氢-4-氧 代 -3-喹 啉 羧 酸：m.p.219.3-222.0 ℃ ,H NMR(600MHz,DMSO-d6)δ:14.34(s,1H,H-11),9.20(s,1H,H-21),8.83(s,1H,H-2),8.13(s,1H,H-20),7.92(d,1H,J=12Hz,H-5),5.76(s,1H,H-18),4.40-4.38(m,1H,H-12),3.48-3.44(m,3H,H-14、H-15and H-17),3.30-3.27(m,4H,H-14、H-15、H-16and H-17),1.22-1.19(m,2H,H-13),1.02-0.99
13
(m,2H,H-13); C NMR(151MHz,DMSO-d6)δ:176.53,165.52,160.13,(156.84,155.18),15
3.25,(143.99,143.90),138.35,(123.42,123.37),(120.17,120.14),(111.09,110.93),108.12,51.07,51.04,44.40(2×C),42.00,11.27(2×C);HR MS C18H19ClFN4O5计算值为
425.1022,实测值为425.1027; (c=1mg/mL,DMF).
[0046] TM2-67-(3-(3-甲氧基脲)-1-吡咯烷基)-1-环丙基-6-氟-8-氯-1,4-二1
氢-4-氧代-3-喹啉羧酸：m.p.252.0-257.0℃,H NMR(600MHz,CDCl3)δ:14.32(s,1H,H-
11),8.92(s,1H,H-2),8.06(d,1H,J=12Hz,H-5),5.30(s,1H,H-18),4.36-4.34(m,1H,H-12),3.78(s,3H,H-21),3.66-3.64(m,3H,H-14、H-15and H-17),3.41-3.39(m,4H,H-14、H-15、
13
H-16and H-17),1.34-1.32(m,2H,H-13),0.99-0.97(m,2H,H-13); C NMR(151MHz,CDCl3)δ:176.77,165.93,157.18,155.50,152.16,(143.89,143.80),137.85,(124.30,
124.24),(120.41,120.39),(112.06,111.91),108.74,64.29,50.90,47.33,44.4
9,41.30,29.70,11.49(2×C);HR MS C19H21ClFN4O5计 算 值 为 439.1179, 实 测 值 为
439.1175; (c=1mg/mL,DMF).
氢-4-氧代-3-喹啉羧酸：m.p.252.0-257.0℃,H NMR(600MHz,CDCl3)δ:14.32(s,1H,H-
11),8.92(s,1H,H-2),8.06(d,1H,J=12Hz,H-5),5.30(s,1H,H-18),4.36-4.34(m,1H,H-12),3.78(s,3H,H-21),3.66-3.64(m,3H,H-14、H-15and H-17),3.41-3.39(m,4H,H-14、H-15、
13
H-16and H-17),1.34-1.32(m,2H,H-13),0.99-0.97(m,2H,H-13); C NMR(151MHz,CDCl3)δ:176.77,165.93,157.18,155.50,152.16,(143.89,143.80),137.85,(124.30,
124.24),(120.41,120.39),(112.06,111.91),108.74,64.29,50.90,47.33,44.4
9,41.30,29.70,11.49(2×C);HR MS C19H21ClFN4O5计 算 值 为 439.1179, 实 测 值 为
439.1175; (c=1mg/mL,DMF).
[0047] TM2-77-(3-(3,3- 二 (2- 羟 乙 基 ) 脲 )-1- 吡 咯 烷 基 )-1- 环 丙1
基 -6- 氟 -8- 氯 -1,4- 二 氢 -4- 氧 代 -3- 喹 啉 羧 酸：m.p.218.1-219.4 ℃ ;H NMR(300MHz,DMSO-d6)δ:14.53(s,1H,H-11),8.82(s,1H,H-2),7.93(d,1H,J=12Hz,H-5),6.63(t,2H,J=6Hz,H-22),4.73(m,1H,H-18),4.39-4.37(m,1H,H-12),3.53-3.48(m,4H,
13
H-21),3.33-3.25(m,11H,H-14、H-15、H-16、H-17and H-20),1.21-0.98(m,4H,H-13); C NMR(75MHz,DMSO-d6)δ:(176.41,176.37),165.43,164.41,157.50,(154.17,153.04),(144.17,143.97),(138.35,138.33),(123.03,122.93),(119.75,119.68),(111.02,110.70),107.98,59.16,51.03,50.99,50.47,47.9(2×C)6,43.44,11.26(2×C);HR MS:C22H26ClFN4NaO6计算值为519.1417,实测值为519.1419; (c=1mg/mL,DCM).[0048] 实施例3、化合物TM3的合成
基 -6- 氟 -8- 氯 -1,4- 二 氢 -4- 氧 代 -3- 喹 啉 羧 酸：m.p.218.1-219.4 ℃ ;H NMR(300MHz,DMSO-d6)δ:14.53(s,1H,H-11),8.82(s,1H,H-2),7.93(d,1H,J=12Hz,H-5),6.63(t,2H,J=6Hz,H-22),4.73(m,1H,H-18),4.39-4.37(m,1H,H-12),3.53-3.48(m,4H,
13
H-21),3.33-3.25(m,11H,H-14、H-15、H-16、H-17and H-20),1.21-0.98(m,4H,H-13); C NMR(75MHz,DMSO-d6)δ:(176.41,176.37),165.43,164.41,157.50,(154.17,153.04),(144.17,143.97),(138.35,138.33),(123.03,122.93),(119.75,119.68),(111.02,110.70),107.98,59.16,51.03,50.99,50.47,47.9(2×C)6,43.44,11.26(2×C);HR MS:C22H26ClFN4NaO6计算值为519.1417,实测值为519.1419; (c=1mg/mL,DCM).[0048] 实施例3、化合物TM3的合成
[0049]
[0050] 向反应瓶中加入氨基被叔丁氧羰基(Boc)保护的氨基酸Boc-AA-OH 12mmol和DCM20mL，搅拌1min，在冰浴下加入1-羟基苯并三氮唑(HOBt)12mmol和二环己基碳化二亚胺(DCC)15mmol，搅拌1min，再滴加TEA 1mL，冰浴下搅拌约30min，至原料全部生成活化酯后，抽滤，滤饼用DCM洗涤，洗液和滤液合并，加入克林沙星10mmol，常温搅拌反应，TLC监测反应进程。反应完毕，抽滤，滤饼用DCM洗涤，洗液和滤液合并，分别用10%(w/w)柠檬酸溶液、
0.5mol/L NaHCO3溶液、饱和NaCl溶液洗涤，收集有机相，无水Na2SO4干燥，旋蒸，柱层析纯化，得中间体IM3。
0.5mol/L NaHCO3溶液、饱和NaCl溶液洗涤，收集有机相，无水Na2SO4干燥，旋蒸，柱层析纯化，得中间体IM3。
[0051] 向反应瓶中加入中间体IM3和适量DCM，室温搅拌溶解，冰浴下滴加TFA(VTFA:VDCM=1:3)，室温搅拌反应，TLC监测反应进程。反应完毕，在通风橱中抽TFA30min，然后尽量旋蒸除去TFA，再把反应液慢慢加入到冰冻乙醚中以析出产品，离心，固体用冷冻乙醚(15mL×2)洗涤，干燥，溶于少量乙醇中，加入10%(w/w)Na2CO3的冰水溶液，搅拌，静置，过滤得固体，再溶于无水乙醇中，加入HCl，搅拌，静置，抽滤，洗涤，干燥，即得化合物TM4。合成结果见表3。
[0052] 表3化合物TM3合成结果
[0053]
[0054] TM3-17-(3-(2-氨基乙酰氨基)-1-吡咯烷基)-1-环丙基-6-氟-8-氯-1,4-二1
氢-4-氧代-3-喹啉羧酸盐酸盐：m.p.223.1-225.6℃;H NMR(300MHz,DMSO-d6)δ:14.48(s,1H,H-11),8.85(s,1H,H-2),8.08(s,3H,H-21),7.97(d,1H,J=11.7Hz,H-5),4.34-4.45(m,1H,H-12),3.96(s,2H,H-20),3.49-3.79(m,5H,H-14,H-16and H-17),1.71-2.14(m,2H,H-
13
15),0.97-1.22(m,4H,H-13); CNMR(75MHz,DMSO-d6)δ:11.28,39.64,39.81,40.18,40.46,42.01,51.02,108.13,111.12,120.38,123.42,133.68,138.25,143.81,153.27,165.41,1+
67.63,176.54;HR MS:C19H20ClFN4O4[M+H] 计算值为423.1230,测定值为423.1232.[0055] TM3-27-(3-(2- 吡 咯 烷 基 甲 酰 氨 基 )-1- 吡 咯 烷 基 )-1- 环 丙
1
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸盐酸盐：m.p.196.8-199.8℃;H NMR(300MHz,DMSO-d6)δ:14.47(s,1H,H-11),8.85(s,1H,H-2),8.14(s,2H,H-24,),7.97(d,1H,J=11.7Hz,H-5),4.32-4.50(m,2H,H-12and H-20),3.60-3.80(m,6H,H-14,H-17and H-23),3.38-3.53(m,1H,H-16),2.04-2.17(m,2H,H-21),1.68-2.00(m,2H,H-15),1.33-1.
13
35(m,2H,H-22),0.88-1.31(m,4H,H-13); C NMR(75MHz,DMSO-d6)δ:11.27,24.25,28.76,33.54,39.64,40.47,41.88,46.72,51.34,62.04,108.09,111.20,123.64,128.36,133.+
80,138.28,143.70,153.28,165.48,168.69,176.64;HR MS:C22H24ClFN4O4[M+H] 计算值为
463.1543,测定值为463.1540.
氢-4-氧代-3-喹啉羧酸盐酸盐：m.p.223.1-225.6℃;H NMR(300MHz,DMSO-d6)δ:14.48(s,1H,H-11),8.85(s,1H,H-2),8.08(s,3H,H-21),7.97(d,1H,J=11.7Hz,H-5),4.34-4.45(m,1H,H-12),3.96(s,2H,H-20),3.49-3.79(m,5H,H-14,H-16and H-17),1.71-2.14(m,2H,H-
13
15),0.97-1.22(m,4H,H-13); CNMR(75MHz,DMSO-d6)δ:11.28,39.64,39.81,40.18,40.46,42.01,51.02,108.13,111.12,120.38,123.42,133.68,138.25,143.81,153.27,165.41,1+
67.63,176.54;HR MS:C19H20ClFN4O4[M+H] 计算值为423.1230,测定值为423.1232.[0055] TM3-27-(3-(2- 吡 咯 烷 基 甲 酰 氨 基 )-1- 吡 咯 烷 基 )-1- 环 丙
1
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸盐酸盐：m.p.196.8-199.8℃;H NMR(300MHz,DMSO-d6)δ:14.47(s,1H,H-11),8.85(s,1H,H-2),8.14(s,2H,H-24,),7.97(d,1H,J=11.7Hz,H-5),4.32-4.50(m,2H,H-12and H-20),3.60-3.80(m,6H,H-14,H-17and H-23),3.38-3.53(m,1H,H-16),2.04-2.17(m,2H,H-21),1.68-2.00(m,2H,H-15),1.33-1.
13
35(m,2H,H-22),0.88-1.31(m,4H,H-13); C NMR(75MHz,DMSO-d6)δ:11.27,24.25,28.76,33.54,39.64,40.47,41.88,46.72,51.34,62.04,108.09,111.20,123.64,128.36,133.+
80,138.28,143.70,153.28,165.48,168.69,176.64;HR MS:C22H24ClFN4O4[M+H] 计算值为
463.1543,测定值为463.1540.
[0056] TM3-37-(3-(2-氨基-4-(甲 硫基)丁 酰氨 基)-1-吡咯 烷基)-1-环 丙1
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸盐酸盐：m.p.244.7-247.6℃;H NMR(300MHz,DMSO-d6)δ:14.27(s,1H,H-11),8.92(s,1H,H-2),8.76(s,3H,H-24),8.0
6(d,1H,J=11.1Hz,H-5),5.47(t,1H,J=9Hz,H-20),4.31-4.39(m,1H,H-12),4.03(t,2H,J=9Hz,H-22),3.52-3.92(m,4H,H-14and H-17),3.44-3.53(m,1H,H-16),2.43(s,3H
13
,H-23),2.17-2.21(m,2H,H-21),1.68-2.03(m,2H,H-15),0.91-1.36(m,4H,H-13); C NMR(75MHz,DMSO-d6)δ:11.41,15.19,28.39,33.89,34.39,41.29,43.10,48.82,49.20,51.13,107.96,111.19,123.24,125.48,134.66,138.42,143.79,153.26,165.47,167.63,176.54.
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸盐酸盐：m.p.244.7-247.6℃;H NMR(300MHz,DMSO-d6)δ:14.27(s,1H,H-11),8.92(s,1H,H-2),8.76(s,3H,H-24),8.0
6(d,1H,J=11.1Hz,H-5),5.47(t,1H,J=9Hz,H-20),4.31-4.39(m,1H,H-12),4.03(t,2H,J=9Hz,H-22),3.52-3.92(m,4H,H-14and H-17),3.44-3.53(m,1H,H-16),2.43(s,3H
13
,H-23),2.17-2.21(m,2H,H-21),1.68-2.03(m,2H,H-15),0.91-1.36(m,4H,H-13); C NMR(75MHz,DMSO-d6)δ:11.41,15.19,28.39,33.89,34.39,41.29,43.10,48.82,49.20,51.13,107.96,111.19,123.24,125.48,134.66,138.42,143.79,153.26,165.47,167.63,176.54.
[0057] 实施例4、化合物TM4的合成
[0058]
[0059] 向反应瓶中依次加入Boc-Met-OH、HOBt和适量THF，搅拌使溶解，冰浴冷却，加入DCC和二异丙基乙二胺(DIPEA)，冰浴搅拌30分钟，加入CF并补加适量THF，CF、Boc-Met-OH、HOBt、DCC和DIPEA的投料摩尔比为10:14:12-15:12-15:15-20，室温搅拌反应，TLC监测反应进程。反应完毕，将反应液于4℃静置过夜，抽滤，滤饼用DCM洗涤，洗液与滤液合并，减压蒸馏除去溶剂，残余物用DCM溶解后，依次用0.5mol/L NaHCO3溶液、10%(w/w)柠檬酸溶液和饱和食盐水洗涤，收集有机相，无水Na2SO4干燥，过滤，滤液减压蒸馏除去溶剂，用DCM-石油醚混合溶剂重结晶，得中间体IM4-1。
[0060] 向反应瓶中加入中间体IM4-11mmol，再逐步加入DCM4mL，冰浴冷却，加入间氯过氧苯甲酸(mCPBA)2.1mmol，0℃搅拌反应，TLC监测反应进程。反应完毕，如得淡黄色浑浊液，则补加适量DCM使固体完全溶解；如得澄清液体，则不需补加DCM；然后依次用5%(w/w)Na2CO3溶液和饱和食盐水洗涤，无水Na2SO4干燥，过滤，滤液减压蒸馏除去溶剂，用DCM-PE混合溶剂重结晶，得中间体IM4-2。
[0061] 向反应瓶中加入中间体IM4-21mmol和DCM3～6mL，搅拌溶解，冰浴冷却，缓慢滴加TFA1～3mL，冰浴搅拌反应，TLC监测反应进程。反应完毕，减压蒸馏除去TFA，残液慢慢加入冰冻乙醚中，搅拌，冰箱中冷藏析晶，吸弃乙醚，固体用冷冻乙醚洗涤，干燥，室温下缓慢滴加饱和K2CO3溶液15～25mL(以原料完全溶解为准)，用稀盐酸调节pH至3～4，如有固体析出，则抽滤，滤饼用少量水洗涤，得粗品；如无固体析出，则向溶液中加入固体食盐至达到饱和，再置冰箱中冷藏析晶，抽滤，滤饼用少量水洗涤，得粗品。将粗品用92%(v/v)乙醇分散，抽滤，干燥，得化合物TM4。
[0062] TM47-(3-(2-氨基-4- 甲磺 酰基)丁 酰氨 基)-1-吡 咯烷 基)-1-环 丙1
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸盐酸盐:m.p.222.9-225.5℃;H NMR(300MHz,DMSO-d6)δ:14.35(s,1H,H-11),8.85(s,1H,H-2),8.55(s,3H,H-24),7.98(d,1H,J=11.4Hz,H-5),4.60-4.63(m,1H,H-16),4.40-4.43(m,1H,H-12),3.64-3.81(m,4H,H-14and H-17),3.56(m,1H,H-20),3.36-3.37(m,2H,H-22),3.33-3.35(m,2H,H-21),3.0
13
5(s,3H,H-23),2.19-2.21(m,2H,H-15),0.98-1.21(m,4H,H-13); C NMR(75MHz,DMSO-d6)δ:11.32(2×C),24.31,40.72,42.01,42.99,46.24,48.48,49.58,50.81,51.26,107.
96,110.85,120.13,123.27,138.23,143.58,143.77,153.11,165.38,166.99,176.47;
+
HR MS:C22H27ClFN4O6S[M+H] 计算值为529.1318,实测值为529.1314; (c=1mg/mL,H2O).
基-6-氟-8-氯-1,4-二氢-4-氧代-3-喹啉羧酸盐酸盐:m.p.222.9-225.5℃;H NMR(300MHz,DMSO-d6)δ:14.35(s,1H,H-11),8.85(s,1H,H-2),8.55(s,3H,H-24),7.98(d,1H,J=11.4Hz,H-5),4.60-4.63(m,1H,H-16),4.40-4.43(m,1H,H-12),3.64-3.81(m,4H,H-14and H-17),3.56(m,1H,H-20),3.36-3.37(m,2H,H-22),3.33-3.35(m,2H,H-21),3.0
13
5(s,3H,H-23),2.19-2.21(m,2H,H-15),0.98-1.21(m,4H,H-13); C NMR(75MHz,DMSO-d6)δ:11.32(2×C),24.31,40.72,42.01,42.99,46.24,48.48,49.58,50.81,51.26,107.
96,110.85,120.13,123.27,138.23,143.58,143.77,153.11,165.38,166.99,176.47;
+
HR MS:C22H27ClFN4O6S[M+H] 计算值为529.1318,实测值为529.1314; (c=1mg/mL,H2O).
[0063] 实施例5、化合物TM5的合成
[0064]
[0065] 向反应瓶中加入5mL醇和足量钠片，室温搅拌，待无气泡产生时，取出多余钠片，根据醇钠的状态加入适量相应醇和化合物A1mmol，继续室温搅拌反应，TLC监测反应进程。反应完毕，用3N HCl调pH=4～5，加入10mL DCM萃取，有机相用水(2×10mL)洗涤，无水Na2SO4干燥。对于沸点较低的醇，旋蒸得粗品；对于沸点高、水溶性不好的醇，旋蒸浓缩除去DCM，加入乙醚，往往有大量固体析出，抽滤，滤饼用乙醚洗涤，得粗品。粗品用柱层析纯化，即得化合物TM5。
[0066] 表4化合物TM5合成结果
[0067]
[0068] TM5-17-(3-((甲氧羰基)氨基)-1-吡咯烷基)-1-环丙基-6-氟-8-氯-1,4-二1
氢-4-氧代-3-喹啉羧酸：m.p.205.6-208.2℃,H NMR(600MHz,CDCl3)δ:14.34(s,1H,H-
11),8.92(s,1H,H-2),8.05(d,1H,J=12Hz,H-5),5.30(s,1H,H-18),4.38-4.34(m,1H,H-12),3.76(s,3H,H-20),3.69-3.67(m,3H,H-14、H-15and H-17),3.34-3.32(m,4H,H-14、H-15、
13
H-16and H-17),1.33-1.25(m,2H,H-13),1.00-0.97(m,2H,H-13); C NMR(151MHz,CDCl3)δ:(176.78,176.75),165.93,157.59,(155.94,155.07),152.07,(144.21,144.07),137.
86,(124.10,124.02),(120.24,120.19),(112.01,111.17),108.68,52.84,51.04,51.00,-
44.39,41.27,29.68,11.45(2×C);HR MS calcd for C19H19ClFN3O5[M-H]422.0925,found422.0928; (c=1mg/mL,DCM).
氢-4-氧代-3-喹啉羧酸：m.p.205.6-208.2℃,H NMR(600MHz,CDCl3)δ:14.34(s,1H,H-
11),8.92(s,1H,H-2),8.05(d,1H,J=12Hz,H-5),5.30(s,1H,H-18),4.38-4.34(m,1H,H-12),3.76(s,3H,H-20),3.69-3.67(m,3H,H-14、H-15and H-17),3.34-3.32(m,4H,H-14、H-15、
13
H-16and H-17),1.33-1.25(m,2H,H-13),1.00-0.97(m,2H,H-13); C NMR(151MHz,CDCl3)δ:(176.78,176.75),165.93,157.59,(155.94,155.07),152.07,(144.21,144.07),137.
86,(124.10,124.02),(120.24,120.19),(112.01,111.17),108.68,52.84,51.04,51.00,-
44.39,41.27,29.68,11.45(2×C);HR MS calcd for C19H19ClFN3O5[M-H]422.0925,found422.0928; (c=1mg/mL,DCM).
[0069] TM5-27-(3-((异丙氧羰基)氨基)-1-吡咯烷基)-1-环丙基-6-氟-8-氯-1,4-二1
氢-4-氧代-3-喹啉羧酸：m.p.191.8-193.3℃,H NMR(600MHz,CDCl3)δ:14.34(s,1H,H-
11),8.91(s,1H,H-2),8.04(d,1H,J=12Hz,H-5),5.30(s,1H,H-18),5.01-4.95(m,1H,H-20),4.37-4.35(m,1H,H-12),3.69-3.67(m,3H,H-14、H-15and H-17),3.36-3.34(m,4H,H-14、H-15、H-16and H-17),1.34-1.30(m,2H,H-13),1.29(s,3H,H-21),1.28(s,3H,H-21),0.99-
13
0.96(m,2H,H-13); C NMR(151MHz,CDCl3)δ:(176.77,176.75),165.95,157.56,(155.19,
155.04),152.03,(144.30,144.16),137.87,(124.00,123.96),(120.11,120.06),(111.99,111.76),108.67,69.03,51.09,51.05,44.28,44.19,41.26,22.24(2×C),11.44(2×C);H-
R MS calcd for C21H23ClFN3O5[M-H]450.1238,found450.1378; (c=1mg/mL,DCM).[0070] 实施例6、合成化合物对结核分枝杆菌的最低抑菌浓度(MIC)测定
氢-4-氧代-3-喹啉羧酸：m.p.191.8-193.3℃,H NMR(600MHz,CDCl3)δ:14.34(s,1H,H-
11),8.91(s,1H,H-2),8.04(d,1H,J=12Hz,H-5),5.30(s,1H,H-18),5.01-4.95(m,1H,H-20),4.37-4.35(m,1H,H-12),3.69-3.67(m,3H,H-14、H-15and H-17),3.36-3.34(m,4H,H-14、H-15、H-16and H-17),1.34-1.30(m,2H,H-13),1.29(s,3H,H-21),1.28(s,3H,H-21),0.99-
13
0.96(m,2H,H-13); C NMR(151MHz,CDCl3)δ:(176.77,176.75),165.95,157.56,(155.19,
155.04),152.03,(144.30,144.16),137.87,(124.00,123.96),(120.11,120.06),(111.99,111.76),108.67,69.03,51.09,51.05,44.28,44.19,41.26,22.24(2×C),11.44(2×C);H-
R MS calcd for C21H23ClFN3O5[M-H]450.1238,found450.1378; (c=1mg/mL,DCM).[0070] 实施例6、合成化合物对结核分枝杆菌的最低抑菌浓度(MIC)测定
[0071] 挑取在改良罗氏培养基中传代2周后生长良好的菌株适量，用磨菌器研磨成-3菌悬液，用比浊法调节浓度为1mg/mL，再用通用增菌培养基逐级稀释至1×10 mg/mL及-4
5×10 mg/mL的工作浓度。将待测化合物溶于二甲亚砜(DMSO)中制成5mg/mL的储备液，-20℃保存；临用前用通用增菌培养基稀释为42μg/mL的基础液，再将基础液用通用增菌培养基稀释至不同的实验浓度(4、2、1、0.5、0.25、0.125、0.0625、0.0313、0.0156μg/mL)。在96孔板中分别按10μL/孔加入不同浓度的待测化合物，以氧氟沙星和莫西沙星为阳性药物对照，生长对照孔加入通用增菌培养基，再将不同浓度的菌悬液分别以200μL/孔接种到各孔中，同一块板各孔中的菌种类及菌浓度相同，并设1块复板，然后用胶带将96孔板盖密封，37℃孵育。孵育约7天后，即生长对照孔有大量菌生长且阳性药物抑菌梯度明显时，每孔加入20μL Alamar blue溶液与50μL无菌5%Tween80溶液的混合液，37℃孵育
24小时，记录各孔颜色：蓝色孔为无生长；粉红色孔为有生长；紫红色孔再继续于37℃培养
24小时，不变为粉红色，且相邻的蓝色孔仍为蓝色，记录为有生长。MIC结果见表5和表6。
5×10 mg/mL的工作浓度。将待测化合物溶于二甲亚砜(DMSO)中制成5mg/mL的储备液，-20℃保存；临用前用通用增菌培养基稀释为42μg/mL的基础液，再将基础液用通用增菌培养基稀释至不同的实验浓度(4、2、1、0.5、0.25、0.125、0.0625、0.0313、0.0156μg/mL)。在96孔板中分别按10μL/孔加入不同浓度的待测化合物，以氧氟沙星和莫西沙星为阳性药物对照，生长对照孔加入通用增菌培养基，再将不同浓度的菌悬液分别以200μL/孔接种到各孔中，同一块板各孔中的菌种类及菌浓度相同，并设1块复板，然后用胶带将96孔板盖密封，37℃孵育。孵育约7天后，即生长对照孔有大量菌生长且阳性药物抑菌梯度明显时，每孔加入20μL Alamar blue溶液与50μL无菌5%Tween80溶液的混合液，37℃孵育
24小时，记录各孔颜色：蓝色孔为无生长；粉红色孔为有生长；紫红色孔再继续于37℃培养
24小时，不变为粉红色，且相邻的蓝色孔仍为蓝色，记录为有生长。MIC结果见表5和表6。
[0072] 表5合成化合物对结核分枝杆菌标准敏感株的MIC(μg/mL)
[0073]
[0074] 注：“－”表示未测定。
[0075] 由表5可以看出，整体上，受试化合物对结核分枝杆菌标准敏感株都具有一定的抑制效果，其中，化合物TM1-4和TM1-10的抑菌活性最强，与克林沙星相当，强于氧氟沙星而略弱于莫西沙星；TM1-5的抑菌活性与氧氟沙星相当；TM1-3和TM1-9的抑菌活性略弱于氧氟沙星。此外，化合物的抑菌效果基本不会随着菌株浓度增加而变化。
[0076] 表6部分合成化合物对结核分枝杆菌不同临床分离株的MIC(μg/mL)[0077]
[0078]
[0079] 注：“－”表示未测定。
[0080] 由表6可以看出，整体上，受试化合物对结核分枝杆菌临床分离敏感株和临床分离耐药株都具有一定的抑制效果，其中，化合物TM1-4的抑菌活性最强，对临床分离敏感株的抑菌活性强于氧氟沙星而略弱于莫西沙星，并较克林沙星高2～4倍，对临床分离耐药株1288号也表现出较强的抑菌活性；TM1-5和TM1-10的抑菌活性次之。
[0081] 最后说明的是，以上优选实施例仅用以说明本发明的技术方案而非限制，尽管通过上述优选实施例已经对本发明进行了详细的描述，但本领域技术人员应当理解，可以在