CN112110898A - 辛弗林磺酰化衍生物及其中间体、制备方法和应用 - Google Patents
辛弗林磺酰化衍生物及其中间体、制备方法和应用 Download PDFInfo
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Abstract
本发明公开了辛弗林磺酰化衍生物及其中间体、制备方法和应用,属于药物合成技术领域。辛弗林磺酰化衍生物的结构式如下。体外抗细菌活性测试结果表明,所有化合物均具有抗细菌活性,且对金黄色葡萄球菌显示强的抑制活性,远强于临床药物;同时,制得的中间体对金黄色葡萄球菌的MIC值为0.4μg/mL,强于临床药物。抗柑橘溃疡病菌活性测试结果表明,绝大多数目标化合物对柑橘溃疡病的抑制活性强于辛弗林,多数目标化合物对柑橘溃疡病的抑制活性强于对照药物诺氟沙星。这些结果表明辛弗林磺酰化衍生物及其中间体在抗细菌、抗柑橘溃疡病菌领域具有潜在的应用前景。
Description
技术领域
本发明涉及药物合成技术领域,具体涉及辛弗林磺酰化衍生物及其中间体、制备方法和应用。
背景技术
辛弗林(synephrine),是一种存在于芸香科柑橘属植物中的生物碱,其结构与肾上腺素及麻黄碱相似且具有类似的生物活性。辛弗林主要表现为肾上腺素α受体兴奋剂作用,对心脏β受体也有一定的兴奋作用,能促进血管收缩、提高心血输出量、升高血压、扩张支气管和气管;能促进新陈代谢、燃烧脂肪,从而具有减肥功效。中医临床上,辛弗林主要用于治疗支气管哮喘及低血压、虚脱、休克、体位性低血压,以及治疗食积不化、化痰除痞、胃下垂等病症;辛弗林注射液也用于抢救各种休克、心力衰竭病人以及治疗胃和十二指肠溃疡等病症;在中轻度抑郁症治疗、血糖调节等方面,辛弗林也取得了良好疗效。由于毒副作用,麻黄碱近年来受到一些国家禁用,但辛弗林副作用小且可用作麻黄碱的替代品,因此市场前景看好。受限于辛弗林的提取效率不高、反应位点复杂难控,目前对辛弗林的研究以化学合成方法、提取方法改进及其分析测试方面为主,对辛弗林衍生物的合成及生物活性研究较少,影响了辛弗林的发展前景。我国作为柑橘生产大国,每年产出的柑橘皮渣超过1000万吨,含有辛弗林5.5万吨以上,但绝大部分资源被浪费,因此研究开发以辛弗林为母核的衍生物及其生物活性,具有重要的理论意义和显著的经济价值。
致病菌的扩散及其耐药性的增强严重威胁着人类的健康和生命,抗菌药物已作为常规用药广泛用于艾滋病、器官移植以及慢性消耗性疾病(如癌症、糖尿病、尿毒症等)的治疗,虽然目前临床上使用的抗菌药剂(如酮康唑、阿米卡星、庆大霉素、活力康唑、伊曲康唑、特比萘芬、二性霉素、氟康唑等)对皮肤及浅表部位感染的疗效较好,但这些抗菌药物的蓄积毒性较强,常常引起肝肾损伤、消化道刺激、头晕、过敏等,所以寻找作用机理独特的新型抗菌药物成为当今药物研发的热点之一。
柑橘溃疡病分布广泛,可危害几十种芸香科植物,是影响世界柑橘生产的重大检疫性病害。其危害从柑橘叶、枝以及柑橘果实均有涉及,典型的症状是形成溃疡斑,不及时治疗,病害加重,除了引起植被生长外,还严重危害柑橘生产及经济效益。柑橘溃疡病菌系分化复杂、发病率高、传播快、寄主范围广,所以如何防治柑橘溃疡病一直是一个世界性难题,目前尚无一种方法可以根治。生产时常用波尔多液等含有金属铜离子的混合液体进行杀菌,需多次大量喷洒使用,既可能加速耐药性的产生,还会对土壤、其他益生菌产生毒害。开发新型抗柑橘溃疡病药物迫在眉睫。
发明内容
有鉴于此,本发明的目的在于提供辛弗林磺酰化衍生物及其中间体、制备方法和应用。
经研究,本发明提供以下技术方案:
1、式Ⅰ所示的辛弗林磺酰化衍生物,
R1、R2、R3和R4独立地选自为H、卤素或C1-C3烷基;
R5、R6、R7独立地选自为H或CR8,R8为卤素;
R9为硝基或OCR10;R10为卤素。
R1、R2、R3和R4独立地选自为H、卤素或甲基;
R5、R6、R7独立地选自为H或CR8,R8为F3、Cl3或Br3;
R9为硝基或OCR10;R10为F3。
R5、R6、R7独立地选自为H或CR8,R8为F3、Cl3或Br3;
R9为硝基或OCR10;R10为F3。
优选的,所述式Ⅰ所示的辛弗林磺酰化衍生物为以下化合物中的任一种:
2、上述辛弗林磺酰化衍生物的制备方法,包括以下步骤:
将辛弗林进行氨基保护,制得中间体IMa;
将中间体IMa与linker试剂反应,制得中间体IMb;
将中间体IMb与克林沙星偶联,制得中间体IMc;
将中间体IMc脱除氨基保护基,制得中间体IMd;
将中间体IMd与芳香磺酰氯偶联,制得辛弗林磺酰化衍生物;
IMa、IMb和IMc中B为叔丁氧羰基(Boc)、芴甲氧羰基(Fmoc)或苄氧羰基(Cbz或Z);
IMb中R11为卤素;
式中,L和R的定义与上述的辛弗林磺酰化衍生物结构式中的L和R的定义相同。
优选的,包括以下步骤:
将辛弗林在碱作用下与Boc2O反应,制得中间体IMa;所述有机溶剂为Boc2O;
将中间体IMa和与linker试剂在二氯甲烷和碱作用下偶联,制得中间体IMb;所述碱为碳酸钠、碳酸钾、碳酸氢钠或碳酸氢钾;
将中间体IMb与克林沙星在有机溶剂和碱作用下偶联,制得中间体IMc;所述有机溶剂为二甲基亚砜或N,N-二甲基甲酰胺;所述碱为碳酸钠、碳酸钾、碳酸氢钠或碳酸氢钾;
将中间体IMc在酸性有机试剂作用下脱除氨基保护基,制得中间体IMd;所述酸性有机试剂为乙酸乙酯-氯化氢、乙醚-氯化氢、三氟醋酸或三氟醋酸与有机溶剂的组合物;
将中间体IMd与芳香磺酰氯(RSO2Cl)在有机溶剂、碱性作用下偶联,制得辛弗林磺酰化衍生物;所述有机溶剂为二氯甲烷,所述碱为碳酸钠、碳酸钾、碳酸氢钠或碳酸氢钾。
5、上述辛弗林磺酰化衍生物在抗细菌药物中的应用。
优选的,所述辛弗林磺酰化在抗金黄色葡萄球菌药物中的应用。
优选的,所述辛弗林磺酰化衍生物在抗藤黄微球菌药物中的应用。
6、上述辛弗林磺酰化衍生物在抗柑橘溃疡病菌药物中的应用。
除另有说明外,本发明中的术语“消旋体”是指由等量对映体构成的光学不活性的有机物。“立体异构体”是指原子组成及键接相同而原子在三维空间排列上不同的分子。“药学上可接受的盐”可以是酸性盐,也可以是碱性盐,例如无机酸盐、有机酸盐、无机碱盐或有机碱盐。
术语“C1-C3烷基”指具有1-3个碳原子的直链或支链饱和一价烃基,例如甲基、乙基、丙基和异丙基。
术语“卤素”指F、Cl、Br和I。
本发明的有益效果在于:
1)本发明提供的辛弗林磺酰化衍生物,以辛弗林为母核,对其氨基和酚羟基进行合理修饰,构建了一类结构新颖的辛弗林磺酰化衍生物,产物的化学结构经1H NMR,13C NMR和HR MS确认;
2)化合物对6种细菌的体外抑制活性测试结果表明,所有分子整体对金黄色葡萄球菌保留了高活性,其MIC值在0.2~32μg/mL,有11个分子MIC≤2μg/mL,有8个分子MIC≤0.8μg/mL,远强于临床药物诺氟沙星32μg/mL,与洛美沙星0.8μg/mL、万古霉素1.6μg/mL、环丙沙星2μg/mL、依诺沙星1.6μg/mL活性相当;TM9-17对藤黄微球菌最优抑制浓度为16μg/mL,活性是诺氟沙星的16倍、依诺沙星的4倍,从而证明了辛弗林磺酰化衍生物在抗细菌领域具有潜在的应用前景;
3)本发明提供的辛弗林磺酰化衍生物的制备方法中,制得的中间体IMc与IMd对金黄色葡萄球菌的MIC值为0.4μg/mL,是诺氟沙星的1/80、万古霉素及依诺沙星的1/4、环丙沙星的1/5、洛美沙星的1/2。同时,中间体IMc与IMd还具有广谱性,其中IMd对沙门氏菌的MIC值为2μg/mL,是诺氟沙星的1/32,是依诺沙星的1/8,是环丙沙星、头孢噻吩及洛美沙星的1/4,证明了中间体IMc与IMd在抗细菌领域具有潜在的应用前景;
4)化合物抗柑橘溃疡病菌测试结果表明,所测试绝大部分辛弗林磺酰化衍生物的活性都强于辛弗林。在1.6μg/mL测试浓度下,14个测试分子中,TM9-18的活性强于阳性对照药物诺氟沙星,TM9-1的活性和阳性对照药物诺氟沙星相当,还有9个分子的活性达到阳性对照药物诺氟沙星的80%。在0.64μg/mL浓度下,14个测试分子中,有9个分子的活性强于诺氟沙星。这些结果表明,所测试的绝大多数辛弗林磺酰化衍生物的抗柑橘溃疡病的活性强于辛弗林,多数辛弗林磺酰化衍生物的抗柑橘溃疡病的活性强于阳性对照药物诺氟沙星。从而证明了不少辛弗林磺酰化衍生物在抗柑橘溃疡病药物的开发中具有很好的前景。
具体实施方式
下面结合本发明实施例,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
一、主要试剂和仪器
辛弗林;4,6-二氯嘧啶、Boc2O(AR);乙酸酐(AR);N,N-二异丙基乙胺(DIPEA,AR);碳酸钾(AR)、4-甲基苯磺酰氯(AR)、对氟苯磺酰氯、对氯苯磺酰氯、2,4,6-三甲基苯磺酰氯、苯磺酰氯、邻氟苯磺酰氯、对三氟甲基苯磺酰氯、吡啶-3-磺酰氯、2-(三氟甲氧基)苯磺酰氯、邻氯苯磺酰氯、2,6-二氟苯磺酰氯、3-氟苯磺酰氯(≥97%),邻溴苯磺酰氯(≥98%)、2-(三氟甲基)苯磺酰氯、邻硝基苯磺酰氯、3-溴苯磺酰氯(≥98%),2-噻吩磺酰氯、3-(三氟甲基)苯磺酰氯(≥98%),其余试剂均为市售化学纯或分析纯产品。
核磁共振仪(AV-600,TMS为内标);高分辨质谱仪(HR ESI,Q TOF,);熔点测定仪(X-6);自动旋光仪(WZZ-2S);紫外分析仪(ZF-1);旋转蒸发仪(RE-2000)。
二、辛弗林磺酰胺类衍生物的制备方法
1、中间体IMa-1的合成
向反应瓶中依次加入原料辛弗林10mmol、饱和碳酸钠溶液3mL,室温下搅拌。移至冰水浴中,搅拌半小时,缓慢滴加入Boc2O 12mmol。滴毕,搅拌半小时撤去冰水浴,遮光持续搅拌反应。TLC监测下适当补加饱和碳酸钠溶液至反应完全。反应结束后,加入冰冷饱和NaCl溶液搅拌,2N稀盐酸调节pH为6-7,搅拌均匀后移入冰柜冷却20min,抽滤,滤饼用石油醚分散提纯后,真空干燥,得IMa-1(白色固体)5.419g,收率为101.2%。
2、中间体IMb-1的合成
向反应瓶中依次加入IMa-1 10mmol、DCM 15mL、碳酸钾15mmol,室温下搅拌,补加DMF 10滴。室温搅拌半小时,加入4,6-二氯嘧啶12mmol,遮光室温下搅拌反应,TLC监测反应进程至反应结束。在冰水浴下向体系加入15mL H2O和30mL DCM,2N稀盐酸调节pH至2-3,用饱和氯化钠溶液洗涤,收集有机相,无水Na2SO4干燥0.5h,减压旋蒸除去溶剂,用少许DCM恰好溶解,加入大量石油醚搅拌,冰柜冷藏1h,旋蒸,加入溶剂30mL(PE/DCM=10/1,v/v)搅拌2h,抽滤,TLC监测,滤液含4,6-二氯嘧啶,滤饼经过反复分散洗涤后为纯品,真空干燥,得IMb-1(白色固体粉末)6.771g,收率为89.2%。
3、中间体IMc-1的合成
向反应瓶中依次加入克林沙星(CLX,5.0mmol)、盐酸、DMSO(8mL),搅拌溶解,加入碾碎烘干的K2CO3(15.0mmol),室温下搅拌0.5h,加入IMb(6.0mmol),加入KI(0.2mmol),45℃下搅拌,TLC监测至反应结束。冷却至室温,依次加入冰冷饱和NaCl溶液30mL和DCM50mL,搅拌,2N稀盐酸调节pH至2-3,分液,TLC监测下适当反萃,用饱和氯化钠溶液(30mL×3)洗涤,无水硫酸钠干燥2h,旋蒸除去溶剂,得到粗产品,柱层析纯化(DCM/CH3OH=80/1~40/1,v/v),收集洗脱液,减压蒸干;TLC检查纯度,真空干燥,低温保存,得IMc-1(黄色固体)4.569g,收率为65.8%。
4、辛弗林磺酰胺类衍生物TM9以及中间体IMd-1的合成
于100mL圆底烧瓶中依次加入IMc-1、DCM(4mL),搅拌,冰浴冷却下滴加入乙酸乙酯-氯化氢溶液。TLC监测,适量补加乙酸乙酯-氯化氢溶液直到反应完全。旋蒸除去溶剂,得到IMd。加入DCM(4mL)、甲醇4滴,搅拌溶解,将其四等分至4个100mL反应瓶中,四个瓶子均再加入碾细的干燥K2CO3,移至-5℃恒温低温反应器中搅拌30min,加入芳香磺酰氯,TLC监测反应进程,当IMd-1基本耗尽时,加入冰冷饱和碳酸钠溶液,淬灭停止反应。移至室温下,依次加入10mL冰冷饱和Na2CO3溶液和45mL DCM,搅拌,分液,TLC监测下,饱和氯化钠30mL×3洗涤,无水硫酸钠干燥2h,旋蒸除去溶剂,柱层析纯化(DCM/CH3OH=100/1~50/1,v/v),收集洗脱液,减压蒸干;TLC检查纯度,真空干燥,低温保存,得目标化合物TM9,实验条件及结果如表1所示。
表1 TM9系列目标化合物合成的条件及结果
注:此步骤收率为脱Boc和磺酰化两步的总收率。
5、产物结构表征数据如下:
目标分子的波谱数据表征
8-Chloro-1-cyclopropyl-6-fluoro-7-(3-((6-(4-((R)-1-hydroxy-2-((N-methyl-4-(trifluoromethyl)phenyl)sulfonamido)ethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-4-oxo-1,4-dihydroquinol ine-3-carboxylic acid(TM9-1),yellow oil;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ14.52(s,1H,H-1),8.85(s,1H,H-2),8.24(s,1H,H-3),7.97(d,J=8.6Hz,5H,H-4and H-5),7.39(d,J=8.5Hz,2H,H-6),7.12(d,J=8.4Hz,2H,H-7),6.36(s,1H,H-9),5.65(d,J=4.4Hz,1H,H-8),4.79(s,1H,H-10),4.40(brs,1H,H-11),3.81(s,4H,H-12,H-13 and H-14),3.40(s,4H,H-15 and H-16),2.81(s,3H,H-17),1.95(d,J=48.0Hz,1H,H-18),1.25–1.15(m,3H,H-18,H-19 and H-20),1.00(q,J=6.8Hz,2H,H-19 and H-20).13C NMR(151MHz,DMSO-d6)δ176.67,170.40,165.55,164.48,158.03,153.31,152.55,144.01,142.04,140.09,138.47,128.52,127.89,127.02,124.97,121.59,116.04,111.05,87.29,71.29,57.49,50.86,45.02,42.01,36.50,14.46,11.28.HR MS calcd for C37H33ClF4N6O7S[M+H]+817.1829,found817.1830.
8-Chloro-1-cyclopropyl-7-(3-((6-(4-((R)-2-((N,4-dimethylphenyl)sulfonamido)-1-hydroxy-ethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carbo-xylic acid(TM9-2),yellow oil;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ14.53(s,1H,H-1),8.85(s,1H,H-2),8.24(s,1H,H-3),7.97(d,J=11.7Hz,1H,H-4),7.63(d,J=8.0Hz,2H,H-5),7.40(dd,J=14.5,8.2Hz,4H,H-6 and H-7),7.11(d,J=8.3Hz,2H,H-8),6.35(s,1H,H-9),5.61(d,J=4.3Hz,1H,H-10),4.78(s,1H,H-11),4.40(brs,1H,H-12),3.81(s,4H,H-13),3.40(s,4H,H-14),3.08(d,J=5.8Hz,2H,H-15),2.72(s,3H,H-16),2.39(s,3H,H-17),1.18(d,J=6.3Hz,2H,H-18),1.00(s,2H,H-18).13C NMR(151MHz,DMSO-d6)δ176.66,170.42,165.54,164.47,158.03,156.91,155.39,153.42,153.30,152.78,152.49,149.67,143.62,140.29,138.47,134.92,130.29,127.89,127.56,123.42,121.57,120.17,111.20,111.04,108.23,87.27,71.47,57.64,50.86,45.01,42.02,36.69,21.43,11.29.HR MS calcd forC37H36ClFN6O7S[M+H]+763.2112,found 763.2113.
8-Chloro-1-cyclopropyl-6-fluoro-7-(3-((6-(4-((R)-2-((4-fluoro-N-methylphenyl)sulfonamido)-1-hydroxyethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-4-oxo-1,4-dihydroquinoline-3-carb oxylic acid(TM9-3),yellowsolid,m.p.>250℃;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ8.63(s,1H,H-1),8.23(s,1H,H-2),7.82(dd,J=12.2,5.9Hz,3H,H-3),7.48–7.36(m,4H,H-4),7.12(d,J=8.3Hz,2H,H-5),6.35(s,1H,H-6),4.78(t,J=6.0Hz,1H,H-7),4.27–4.21(m,1H,H-8),3.75(s,4H,H-9),3.44(s,4H,H-9),3.15–3.08(m,2H,H-10),2.76(s,3H,H-11),1.13(d,J=6.3Hz,2H,H-12),0.92(s,2H,H-12).13C NMR(151MHz,DMSO-d6)δ171.48,170.35,165.73,164.79,164.46,164.02,157.99,155.11,153.20,152.52,142.27,142.15,140.20,137.53,134.35,130.62,130.55,127.89,126.64,126.60,121.58,120.53,120.47,117.05,116.90,111.79,110.24,87.25,71.36,57.55,51.90,50.74,45.06,36.57,11.18.HR MScalcd for C37H35ClF2N6O7S[M+H]+781.2017,found 781.2025.
8-Chloro-7-(3-((6-(4-((R)-2-((4-chloro-N-methylphenyl)sulfonamido)-1-hydroxyethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-ca-rbo xylic acid(TM9-4),yellow oil;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ14.52(s,1H,H-1),8.85(s,1H,H-2),8.24(s,1H,H-3),7.96(d,J=11.6Hz,1H,H-4),7.77(d,J=8.0Hz,2H,H-5),7.67(d,J=8.0Hz,2H,H-6),7.39(d,J=7.9Hz,2H,H-7),7.12(d,J=7.9Hz,2H,H-8),6.36(s,1H,H-9),5.64(d,J=3.7Hz,1H,H-10),4.79(d,J=4.2Hz,1H,H-11),4.40(brs,1H,H-12),3.81(s,4H,H-13),3.40(s,4H,H-13),3.14(d,J=6.2Hz,2H,H-14),2.77(s,3H,H-15),1.19(d,J=6.3Hz,2H,H-16),1.01(s,2H,H-16).13C NMR(151MHz,DMSO-d6)δ176.66,170.40,165.54,164.47,158.02,156.91,155.24,153.29,152.52,143.95,143.86,140.17,138.46,138.21,136.85,129.96,129.47,127.89,123.56,121.58,120.17,111.20,111.04,108.22,87.29,71.33,57.53,50.87,45.01,42.02,40.49,40.35,40.21,40.07,39.93,39.79,39.65,36.55,11.30.HR MS calcd for C36H33Cl2FN6O7S[M+H]+783.1565,found 783.1580.
8-Chloro-1-cyclopropyl-6-fluoro-7-(3-((6-(4-((R)-1-hydroxy-2-((N,2,4,6-tetramethylphenyl)sulfonamido)ethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid(TM9-5),yellowsolid,m.p.150.6-151.4℃;(c=1mg/mL,CH2Cl2).
1H NMR(600MHz,DMSO-d6)δ14.53(s,1H,H-1),8.86(s,1H,H-2),8.24(s,1H,H-3),7.98(d,J=11.7Hz,1H,H-4),7.25(d,J=8.3Hz,2H,H-5),7.10–6.99(m,4H,H-6),6.38(s,1H,H-7),5.56(d,J=4.2Hz,1H,H-8),4.71(d,J=4.6Hz,1H,H-9),4.41(s,1H,H-10),3.82(s,4H,H-11),3.40(s,4H,H-11),3.22(d,J=5.8Hz,2H,H-12),2.75(s,3H,H-13),2.45(s,6H,H-15),2.26(s,3H,H-14),1.20(d,J=6.4Hz,2H,H-16),1.01(s,2H,H-16).13C NMR(151MHz,DMSO-d6)δ176.66,174.30,170.33,165.54,164.48,158.01,156.65,155.86,155.11,153.30,152.51,143.95,142.63,140.35,139.98,138.45,132.82,132.23,127.64,123.42,121.53,120.33,111.05,110.78,108.23,87.35,71.36,56.29,50.86,45.02,42.03,34.69,22.75,20.88,11.30.HR MS calcd for C39H40ClFN6O7S[M+H]+791.2425,found 791.2438.
8-Chloro-1-cyclopropyl-6-fluoro-7-(3-((6-(4-((R)-1-hydroxy-2-(N-methylphenylsulfonamido)ethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid(TM9-6),yellow oil;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ14.56(s,1H,H-1),8.86(s,1H,H-2),8.25(s,1H,H-3),7.99(d,J=11.7Hz,1H,H-4),7.81–7.60(m,5H,H-5),7.40(d,J=8.3Hz,2H,H-6),7.12(d,J=8.3Hz,2H,H-7),6.36(s,1H,H-8),4.79(s,1H,H-9),4.41(brs,1H,H-10),3.82(s,4H,H-11),3.41(s,4H,H-11),3.12(d,J=6.3Hz,2H,H-12),2.75(s,3H,H-13),1.20(d,J=5.4Hz,2H,H-14),1.01(s,2H,H-14).13C NMR(151MHz,DMSO-d6)δ176.64,170.27,165.52,164.40,157.89,153.28,152.46,152.10,143.93,143.83,140.28,140.19,138.45,137.83,137.51,132.70,129.84,128.53,127.90,127.48,121.58,111.19,111.04,110.47,108.21,87.23,71.42,50.85,45.04,42.02,36.87,36.68,11.29.HR MS calcd for C36H34ClFN6O7S[M+H]+749.1955,found 749.1955.
8-Chloro-1-cyclopropyl-6-fluoro-7-(3-((6-(4-((R)-2-((2-fluoro-N-methylphenyl)sulfonamido)-1-hydroxyethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid(TM9-7),yellowsolid,m.p.130.5-131.3℃;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ14.53(s,1H,H-1),8.86(s,1H,H-2),8.25(s,1H,H-3),7.98(d,J=11.7Hz,1H,H-4),7.81(t,J=7.5Hz,1H,H-5),7.73(dd,J=12.6,6.7Hz,1H,H-6),7.49–7.44(m,1H,H-7),7.40(dd,J=16.3,8.0Hz,3H,H-8 and H-9),7.12(d,J=8.3Hz,2H,H-10),6.35(s,1H,H-11),5.76(s,1H,H-12),4.80(dd,J=10.7,5.9Hz,1H,H-13),4.41(brs,1H,H-14),3.79(d,J=32.6Hz,4H,H-15),3.41(s,4H,H-15),3.26(d,J=6.2Hz,2H,H-16),2.85(s,3H,H-17),1.20(d,J=6.7Hz,2H,H-18),1.01(s,2H,H-18).13C NMR(151MHz,DMSO-d6)δ176.66,170.41,165.54,164.46,159.40,158.04,157.73,155.23,153.29,152.51,143.85,140.19,138.45,136.02,131.06,127.81,126.17,125.52,123.43,121.62,120.33,118.03,117.88,111.19,111.04,108.21,87.25,71.59,57.37,55.36,50.83,45.00,42.02,36.32,11.29.HR MScalcd for C36H33ClF2N6O7S[M+H]+767.1861,found 767.1866.
7-(3-((6-(4-((R)-2-((2-Bromo-N-methylphenyl)sulfonamido)-1-hydroxyethyl)phenoxy)pyrimi-din-4-yl)amino)pyrrolidin-1-yl)-8-chloro-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-car boxylic acid(TM9-8),yellow oil;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ14.54(s,1H,H-1),8.86(s,1H,H-2),8.25(s,1H,H-3),7.98(t,J=10.1Hz,2H,H-4),7.87(d,J=7.7Hz,1H,H-5),7.57(td,J=16.8,7.5,1.3Hz,2H,H-6),7.39(d,J=8.4Hz,2H,H-7),7.12(d,J=8.5Hz,2H,H-8),6.36(s,1H,H-9),5.67(s,1H,H-10),4.82(s,1H,H-11),4.41(brs,1H,H-12),3.82(s,4H,H-13),3.43(dd,J=14.3,8.9Hz,6H,H-13),2.89(s,3H,H-14),1.20(d,J=6.7Hz,2H,H-15),1.01(t,J=7.8Hz,2H,H-15).13C NMR(151MHz,DMSO-d6)δ176.68,170.41,165.57,164.47,158.04,153.33,152.37,140.26,138.85,138.53,136.22,134.53,131.67,128.68,127.82,123.15,121.63,119.77,111.13,108.32,87.26,71.98,57.98,50.83,45.01,42.03,36.48,11.29.HR MS calcd for C36H33BrClFN6O7S[M+H]+827.1060,found827.1067.
8-Chloro-1-cyclopropyl-6-fluoro-7-(3-((6-(4-((R)-1-hydroxy-2-((N-methyl-2-(trifluoromethyl)phenyl)sulfonamido)ethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-4-oxo-1,4-dihydro-quinoline-3-carboxylic acid(TM9-9),yellow oil;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ14.54(s,1H,H-1),8.86(s,1H,H-2),8.24(s,1H,H-3),7.99(dd,J=21.9,9.9Hz,3H,H-4),7.86(dt,J=16.2,8.1Hz,2H,H-5 and H-6),7.40(d,J=8.3Hz,2H,H-7),7.12(d,J=8.4Hz,2H,H-8),6.38(s,1H,H-9),5.71(d,J=4.4Hz,1H,H-10),4.85(s,1H,H-11),4.41(brs,1H,H-12),3.79(d,J=33.4Hz,4H,H-13),3.41(s,6H,H-13),2.96(s,3H,H-14),1.20(d,J=6.8Hz,2H,H-15),1.02(s,2H,H-15).13C NMR(151MHz,DMSO-d6)δ176.67,170.36,165.54,164.47,158.02,154.83,153.31,152.55,140.11,138.46,133.92,133.44,130.07,129.18,127.78,126.52,124.06,123.13,121.64,119.70,111.19,108.22,87.30,71.93,57.82,55.35,50.85,45.01,42.03,36.77,31.41,28.94,27.39,26.19,22.93,22.51,18.82,14.39,11.29.HR MS calcd for C37H33ClF4N6O7S[M+H]+817.1829,found 817.1831.
8-Chloro-1-cyclopropyl-6-fluoro-7-(3-((6-(4-((R)-1-hydroxy-2-((N-methyl-2-nitrophenyl)sulfonamido)ethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-4-oxo-1,4-dihydroquino-line-3-carboxylic acid(TM9-10),yellowsolid,m.p.143.0-143.5℃;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ14.54(s,1H,H-1),8.85(s,1H,H-2),8.24(s,1H,H-3),7.97(dd,J=14.5,6.4Hz,3H,H-4),7.87(t,J=7.1Hz,1H,H-6),7.82(dd,J=13.6,6.1Hz,1H,H-5),7.40(d,J=8.4Hz,2H,H-7),7.12(d,J=8.4Hz,2H,H-8),6.35(s,1H,H-9),5.70(s,1H,H-10),4.82(s,1H,H-11),4.40(brs,1H,H-12),3.78(d,J=28.1Hz,4H,H-13),3.40(s,6H,H-13),2.93(s,3H,H-14),1.19(d,J=6.6Hz,2H,H-15),1.00(s,2H,H-15).13C NMR(151MHz,DMSO-d6)δ176.63,170.39,165.53,164.45,158.03,156.89,153.28,152.53,148.30,143.84,140.02,138.44,134.80,132.82,131.24,130.30,127.83,124.66,121.62,111.18,111.02,108.21,87.24,71.52,57.49,50.85,50.76,45.00,42.03,36.50,11.30.HR MS calcd for C36H33ClFN7O9S[M+H]+794.1806,found 794.1815.
8-Chloro-1-cyclopropyl-6-fluoro-7-(3-((6-(4-((R)-1-hydroxy-2-((N-methyl-2-(trifluoromethox y)phenyl)sulfonamido)ethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-4-oxo-1,4-dihydro quinoline-3-carboxylic acid(TM9-11),yellow solid,m.p.142.9-144.1℃;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ14.54(s,1H,H-1),8.85(s,1H,H-2),8.24(s,1H,H-3),8.02–7.91(m,2H,H-4),7.82–7.77(m,1H,H-5),7.64–7.54(m,2H,H-6),7.37(d,J=8.5Hz,2H,H-7),7.11(d,J=8.5Hz,2H,H-8),6.34(s,1H,H-9),5.65(d,J=4.2Hz,1H,H-10),4.79(dd,J=11.5,4.6Hz,1H,H-11),4.40(brs,1H,H-12),3.81(s,4H,H-13),3.40(s,4H,H-13),3.30–3.24(m,2H,H-16),2.86(s,3H,H-14),1.18(q,J=6.8Hz,2H,H-15),1.00(q,J=6.7Hz,2H,H-15).13C NMR(151MHz,DMSO-d6)δ176.65,170.40,165.52,164.45,158.03,156.64,155.23,153.29,152.49,145.41,143.93,140.16,138.45,135.53,131.65,128.08,127.76,123.42,121.59,120.16,119.70,111.18,111.03,108.21,99.99,87.23,71.67,57.42,50.82,45.00,42.01,36.35,11.28.HR MS calcd for C37H33ClF4N6O8S[M+H]+833.1778,found 833.1787.
8-Chloro-7-(3-((6-(4-((R)-2-((2-chloro-N-methylphenyl)sulfonamido)-1-hydroxyethyl)phen-oxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid(TM9-12),yellow oil;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ14.53(s,1H,H-1),8.85(s,1H,H-2),8.24(s,1H,H-3),8.03–7.91(m,2H,H-4),7.69-7.63(m,2H,H-5),7.54(dd,J=10.9,4.3Hz,1H,H-6),7.37(d,J=8.5Hz,2H,H-7),7.11(d,J=8.5Hz,2H,H-8),6.35(s,1H,H-9),5.66(s,1H,H-10),4.80(t,J=6.2Hz,1H,H-11),4.40(brs,1H,H-12),3.81(s,4H,H-13),3.44–3.36(m,6H,H-13),2.88(s,3H,H-14),1.19(d,J=6.7Hz,2H,H-15),1.00(s,2H,H-15).13C NMR(151MHz,DMSO-d6)δ176.65,170.40,165.53,164.45,158.03,153.29,152.49,144.35,143.71,140.22,138.46,137.09,134.59,132.65,131.63,131.19,128.20,127.81,123.69,121.60,120.04,111.19,108.22,87.24,71.88,57.76,50.82,45.01,42.02,36.43,11.29.HR MS calcd for C36H33Cl2FN6O7S[M+H]+783.1565,found 783.1577.
8-Chloro-1-cyclopropyl-7-(3-((6-(4-((R)-2-((2,6-difluoro-N-methylphenyl)sulfonamido)-1-hyd roxyethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid(TM9-13),yellow solid,m.p.130.5-131.5℃;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ14.54(s,1H,H-1),8.85(s,1H,H-2),8.24(s,1H,H-3),7.98(d,J=11.7Hz,1H,H-4),7.76–7.70(m,1H,H-5),7.38(d,J=8.4Hz,2H,H-6),7.30(t,J=9.2Hz,2H,H-7),7.12(d,J=8.4Hz,2H,H-8),6.35(s,1H,H-9),5.67(d,J=4.4Hz,1H,H-10),4.82(dd,J=10.9,6.0Hz,1H,H-11),4.41(brs,1H,H-12),3.81(s,4H,H-13),3.40(s,4H,H-13),3.30(d,J=6.3Hz,2H,H-16),2.91(s,3H,H-14),1.19(q,J=6.4Hz,2H,H-15),1.00(t,J=7.6Hz,2H,H-15).13C NMR(151MHz,DMSO-d6)δ176.65,170.39,165.52,164.45,160.09,158.39,158.03,156.90,155.23,153.28,152.52,143.94,140.08,138.45,135.97,127.77,123.55,121.62,120.20,116.32,114.12,113.97,111.19,111.03,108.21,87.24,71.40,57.25,50.82,45.01,42.02,36.09,11.29.HR MS calcd for C36H32ClF3N6O7S[M+H]+785.1767,found785.1765.
8-Chloro-1-cyclopropyl-6-fluoro-7-(3-((6-(4-((R)-2-((3-fluoro-N-methylphenyl)sulfonamido)-1-hydroxyethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid(TM9-14),yellowsolid,m.p.131.2-132.9℃;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ8.85(s,1H,H-2),8.24(s,1H,H-3),7.98(d,J=11.7Hz,1H,H-4),7.67(td,J=8.1,5.6Hz,1H,H-5),7.60(dd,J=10.4,5.1Hz,2H,H-6),7.58–7.53(m,1H,H-7),7.40(d,J=8.5Hz,2H,H-8),7.12(d,J=8.5Hz,2H,H-9),6.36(s,1H,H-10),4.81–4.75(m,1H,H-11),4.40(brs,1H,H-12),3.81(s,5H,H-13),3.40(s,4H,H-13),3.21–3.13(m,2H,H-14),2.79(s,3H,H-15),1.19(d,J=6.7Hz,2H,H-16),1.00(d,J=2.0Hz,2H,H-16).13C NMR(151MHz,DMSO-d6)δ176.65,170.34,165.52,164.43,163.16,161.51,157.96,156.64,155.23,153.28,152.48,143.93,140.15,138.45,132.21,132.16,127.91,123.78,121.56,120.48,120.34,114.62,114.46,111.19,111.03,108.22,87.25,71.36,57.50,50.82,45.02,42.01,36.61,22.46,13.90,11.29.HR MS calcd for C36H33ClF2N6O7S[M+H]+767.1861,found 767.1873.
7-(3-((6-(4-((R)-2-((3-Bromo-N-methylphenyl)sulfonamido)-1-hydroxyethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-8-chloro-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydroquinoline-3-carboxylic acid(TM9-15),yellow oil;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ8.85(s,1H,H-2),8.24(s,1H,H-3),7.98(s,1H,H-4),7.90(d,J=7.7Hz,2H,H-5),7.77(d,J=7.8Hz,1H,H-6),7.57(t,J=7.8Hz,1H,H-7),7.40(d,J=8.3Hz,2H,H-8),7.12(d,J=8.3Hz,2H,H-9),6.35(s,1H,H-10),4.80–4.76(m,1H,H-11),4.40(brs,1H,H-12),3.81(s,5H,H-13),3.40(s,4H,H-13),3.18(dd,J=13.7,5.8Hz,2H,H-14),2.78(s,3H,H-15),1.18(d,J=6.8Hz,2H,H-16),1.00(s,2H,H-16).13C NMR(151MHz,DMSO-d6)δ176.64,170.36,165.52,164.43,157.97,156.89,155.23,153.28,152.48,143.93,143.86,143.83,140.14,138.45,136.15,132.10,129.72,127.92,126.58,123.50,122.78,121.56,120.19,111.19,111.15,111.03,109.05,108.22,87.23,71.32,57.44,50.85,46.34,45.01,42.02,36.57,11.30.HR MScalcd for C36H33BrClFN6O7S[M+H]+827.1060,found 827.1068.
8-Chloro-1-cyclopropyl-6-fluoro-7-(3-((6-(4-((R)-1-hydroxy-2-((N-methyl-3-(trifluoromethyl)phenyl)sulfonamido)ethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-4-oxo-1,4–dihydroquino line-3-carboxylic acid(TM9-16),yellow oil;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ8.85(s,1H,H-2),8.24(s,1H,H-3),8.09(d,J=7.8Hz,2H,H-4),8.04–7.93(m,2H,H-5),7.87(t,J=7.8Hz,1H,H-6),7.39(d,J=8.5Hz,2H,H-8),7.12(d,J=8.5Hz,2H,H-9),6.35(s,1H,H-10),5.76(s,1H,H-7),4.79(dd,J=7.4,5.1Hz,1H,H-11),4.41(s,1H,H-12),3.81(s,4H,H-13),3.39–3.16(m,6H,H-13 and H-14),2.80(s,3H,H-15),1.18(d,J=6.9Hz,2H,H-16),1.00(s,2H,H-16).13C NMR(151MHz,DMSO-d6)δ176.66,170.38,165.54,164.45,158.01,153.31,152.51,143.99,140.07,139.53,138.16,131.60,131.52,130.00,127.90,123.90,121.57,120.32,119.68,111.19,108.22,87.25,71.20,57.37,55.41,55.13,50.85,45.01,42.01,36.42,31.11,14.79,11.28.HR MS calcd for C37H33ClF4N6O7S[M+H]+817.1829,found817.1826.
8-Chloro-1-cyclopropyl-6-fluoro-7-(3-((6-(4-((R)-1-hydroxy-2-(N-methylpyridine-3-sulfonami do)ethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid(TM9-17),yellow solid,m.p.140.5-141.6℃;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ14.54(s,1H,H-1),8.94(s,1H,H-2),8.88–8.82(m,2H,H-3),8.24(s,1H,H-4),8.19–8.16(m,1H,H-5),7.98(d,J=11.7Hz,1H,H-6),7.65(dd,J=8.0,4.8Hz,1H,H-7),7.40(d,J=8.5Hz,2H,H-8),7.12(d,J=8.4Hz,2H,H-9),6.36(s,1H,H-10),5.63(d,J=4.3Hz,1H,H-11),4.79(dd,J=7.9,4.1Hz,1H,H-12),4.41(brs,1H,H-13),3.81(s,4H,H-14),3.40(s,4H,H-14),3.24–3.15(m,2H,H-15),2.82(s,3H,H-16),1.19(d,J=6.7Hz,2H,H-17),1.00(d,J=1.9Hz,2H,H-17).13C NMR(151MHz,DMSO-d6)δ176.65,170.39,165.54,164.47,158.03,153.78,153.30,152.53,147.83,140.09,138.47,135.58,134.72,127.89,124.88,123.51,121.59,120.21,111.20,111.05,108.22,87.28,71.22,57.40,50.86,45.01,42.03,36.41,11.30.HR MS calcd for C35H33ClFN7O7S[M+H]+750.1907,found 750.1904.
8-Chloro-1-cyclopropyl-6-fluoro-7-(3-((6-(4-((R)-1-hydroxy-2-(N-methylthiophene-2-sulfon-amido)ethyl)phenoxy)pyrimidin-4-yl)amino)pyrrolidin-1-yl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid(TM9-18),yellow oil;(c=1mg/mL,CH2Cl2).1H NMR(600MHz,DMSO-d6)δ8.86(s,1H,H-2),8.26(s,1H,H-3),7.99(dd,J=11.4,6.4Hz,2H,H-4),7.65(dd,J=3.7,1.1Hz,1H,H-5),7.41(d,J=8.5Hz,2H,H-6),7.25(dd,J=4.9,3.8Hz,1H,H-7),7.13(d,J=8.5Hz,2H,H-8),6.36(s,1H,H-9),5.76(s,1H,H-10),4.81(t,J=6.3Hz,1H,H-11),4.41(brs,1H,H-12),3.82(s,4H,H-13),3.41(s,4H,H-13),3.14–3.07(m,2H,H-14),2.79(s,3H,H-15),1.20(q,J=6.7Hz,2H,H-16),1.01(t,J=7.8Hz,2H,H-16).13C NMR(151MHz,DMSO-d6)δ176.64,170.27,165.52,164.41,157.90,156.89,155.23,153.28,152.47,143.93,143.84,140.19,138.45,137.53,133.54,132.69,128.53,127.90,123.56,121.57,120.20,111.19,108.21,87.24,71.42,57.81,55.35,50.85,50.82,45.04,42.02,36.87,11.30.HR MS calcd for C34H32ClFN6O7S2[M+H]+755.1519,found 755.1520.
三、辛弗林磺酰胺类衍生物的活性检测
1、体外抗细菌活性测定
采用微量肉汤稀释法,测定化合物抑制金黄色葡萄球菌(Staphyloccocus aureusATCC 25129)、藤黄微球菌(Micrococcus luteus)、大肠杆菌(Escherichia coli ATCC25922)、鲍曼不动杆菌(Acinetobacter baumannii ATCC 19606)、沙门氏菌(SalmonellaEnteritidis ATCC 13076)和铜绿假单胞菌(Pseudomonas aeruginosa ATCC 27853)的活性(MIC值)。
(1)样品溶液的制备
用万分之一的电子天平在干燥室内精确称取样品3.2mg于2mL PE管中,移液枪向PE管中加入1mL DMSO,溶解为澄清透明液体,配制成3.2mg/mL的溶液,封口膜封口后,冰柜避光保存。溶液用DMSO溶解,部分难溶的化合物使用DMSO/吐温-80=200/1(v/v)以增加溶解度。
(2)待测液的配制
根据待测物的效果或是含量及所需要的体积,计算出待测物所需的量,准确称取,用适宜的溶剂及稀释剂将待测物稀释至所需的浓度。
浓度配制:母液3.2mg/mL=3200μg/mL,再吸取320μL储备液,用培养基稀释至1mL,其稀释液后浓度为1024μg/mL,即为待测液A。
(3)菌悬液的制备
接种保存的菌株于普通液体培养基中,置于37℃恒温摇床活化培养24小时。活化后用培养基稀释成105CFU/mL的菌悬液备用。
(4)加样操作
初筛:无菌条件下,在96孔板每个孔加入培养基50μL。随后在第一排的第一孔、第二孔加配好的待测液A(浓度为1024μg/mL)50μL,经过此二倍稀释后,首孔浓度为512μg/mL,第一孔中加入待测液后用移液枪充分吹打(至少4次以上)使待测物与培养基充分混匀,然后吸取50μL加入第二排,再充分吹打使之与培养基充分混匀,照此重复直至第八排,吸取50μL弃去;此时每列待测物浓度从高至低(从上至下)依次为512,256,128,64,32,16,8,4(单位:μg/mL)。再在96孔板每孔中加入稀释好的菌液50μL,此时每孔待测物浓度即为最终待测物浓度从高至低(从上至下)依次为256,128,64,32,16,8,4,2(单位:μg/mL)。培养:将接种好的96孔板放入37℃恒温培养箱培养16-20h,然后观察和记录结果。结果判定:培养完成后,将96孔板从恒温箱中取出,观察孔内细菌生长情况。在判定结果之前,要确定空白无药对照孔的细菌正常生长、阴性对照孔无细菌生长时结果才有意义。将肉眼观察没有细菌生长的孔中的药物浓度作为该药物对该细菌的MIC。如果出现跳孔现象或两孔结果不一,则需重复试验进行验证。
辛弗林磺酰胺类衍生物对金黄色葡萄球菌、大肠杆菌、沙门氏菌、鲍曼不动杆菌、藤黄微球菌、铜绿假单胞菌的MIC值的测定结果如表2所示。测试均设有空白对照、阴性对照、阳性对照。
表2化合物对六种病菌的抑制活性(MIC,μg/mL)
从表2中分析可知:TM9系列分子整体对金黄色葡萄球菌保留了高活性,其MIC值在0.2~32μg/mL,有11个分子MIC≤2μg/mL,占总分子数目的61%;有8个分子MIC≤0.8μg/mL,占总分子数目的44%,抑制活性强于或相当于临床药物万古霉素、诺氟沙星、环丙沙星、依诺沙星和洛美沙星。TM9系列分子对藤黄微球菌最优MIC为16μg/mL,活性强于诺氟沙星、依诺沙星;TM9-17与环丙沙星、洛美沙星的MIC值相同,但其分子量更大、摩尔浓度更小,等摩尔浓度下其活性应强于环丙沙星、洛美沙星、依诺沙星和万古霉素。此外,中间体IMc、IMd对金黄色葡萄球菌的MIC值为0.4μg/mL,活性强于诺氟沙星、环丙沙星、依诺沙星和万古霉素。IMd对沙门氏菌MIC为2μg/mL,其值与克林沙星者相同,因此其活性超过了母核克林沙星。中间体IMd对藤黄微球菌的MIC值为1.6μg/mL,对大肠杆菌的MIC为8μg/mL,IMd显示出广谱抗菌性质。这些结果证明辛弗林磺酰化衍生物及其中间体在抗细菌领域具有潜在的应用前景。
2、抗柑橘真菌病菌生物活性测定
(1)待测物母液及稀释液的配制
用适宜的溶剂及稀释剂将待测物母液稀释至所需的浓度。样品质量为1.0mg,先配成待测物母液1.0mg/1mL=1.0mg/mL;每种待测物设置2个稀释浓度,0.001mg/mL(即稀释1000倍,1μg/mL)和0.004mg/mL(即稀释250倍,4μg/mL)。
(2)操作
待测物培养基配制:①稀释1000倍的待测物培养基配制:取5μL浓度为1μg/mL待测物稀释液与5mL热PDA培养基在10mL离心管中充分混匀;②稀释250倍的药剂培养基配制:取20μL浓度为4μg/mL待测物稀释液与4980μL热PDA培养基在10mL离心管中充分混匀。
对照组:以不加待测物的PDA培养基和加入咪鲜胺的培养基(稀释1000和稀释250倍)作为对照,分别为空白对照和阳性对照。
接菌:将配置好的待测物培养基倒入24孔板内,每株菌每种待测物每个浓度倒一个孔。挑取28℃培养7d的菌株的菌丝,接种于每孔。
培养:将24孔板放于28℃、光照16h的培养箱内培养48h。
测量:运用十字交叉法测量菌落直径。
计算:抑制率%=(CK菌落直径值-测量菌落直径值)×100%/CK菌落直径值。
筛选:将不同待测物抑制率同咪鲜胺的抑制率进行比较,获得初筛结果。
所有化合物对柑橘胶孢炭疽病菌菌株Co.3、柑橘褐斑病菌菌株Al.6和柑橘溃疡病菌活性初筛测试结果,如表3所示。
表3化合物对柑橘真菌的抑制活性
3、抗柑橘溃疡病菌生物活性测定
测定方法:称取1mg样品于50μL DMSO中溶解,用超纯水定容,获得不同浓度样品母液。取10μL母液于1mL超纯水(0.02%吐温)中作为样品溶液a,然后采用倍比稀释法依次配制不同浓度的样品溶液b,c,d等。
将已在PDA培养基上培养3d的溃疡病菌用5mL LB液体培养基洗下,加至195mL LB液体培养基中,振荡混匀备用。在各2mL离心管中分别加入450μL柑桔溃疡病菌菌液和上述各不同浓度(a~d)样品溶液50μL,使得各混合菌液中样品最终浓度分别为A(1.6μg/mL)、B(0.64μg/mL)、C(0.5μg/mL)、D(0.1μg/mL),28℃、200r·min-1恒温振荡培养14h后测定OD600下各混合菌液OD值并计算抑制率(抑制率%=(OD空白-OD样品)/OD空白×100%)。每个样品每个浓度三次重复。结果如表4所示。
表4部分化合物对柑橘溃疡病菌的抑制活性
表4显示,所测试绝大部分辛弗林磺酰化衍生物的活性都强于辛弗林。在1.6μg/mL测试浓度下,14个测试分子中,TM9-17的活性强于阳性对照药物诺氟沙星,TM9-1的活性和阳性对照药物诺氟沙星相当,还有9个分子的活性达到阳性对照药物诺氟沙星的80%。0.64μg/mL浓度下,14个测试分子中,有9个分子的活性强于诺氟沙星。这些结果表明,所测试的绝大多数辛弗林磺酰化衍生物的抗柑橘溃疡病的活性强于辛弗林,多数辛弗林磺酰化衍生物的抗柑橘溃疡病的活性强于阳性对照药物诺氟沙星。这些结果显示,不少辛弗林磺酰化衍生物在抗柑橘溃疡病药物的开发中具有很好的前景。
以上所述实施例仅是为充分说明本发明而所举的较佳的实施例,本发明的保护范围不限于此。本技术领域的技术人员在本发明基础上所作的等同替代或变换,均在本发明的保护范围之内。本发明的保护范围以权利要求书为准。
Claims (10)
6.根据权利要求5所述辛弗林磺酰化衍生物的制备方法,其特征在于,包括以下步骤:
将辛弗林在碱作用下与Boc2O反应,制得中间体IMa;所述碱为碳酸钠、碳酸氢钠、氢氧化钠、碳酸钾、碳酸氢钾、氢氧化钾;
将中间体IMa与linker试剂在二氯甲烷中、碱作用下偶联,制得中间体IMb;所述碱为碳酸钠、碳酸钾、碳酸氢钠或碳酸氢钾;
将中间体IMb与克林沙星在有机溶剂中、碱作用下偶联,制得中间体IMc;所述有机溶剂为二甲基亚砜或N,N-二甲基甲酰胺;所述碱为碳酸钠、碳酸钾、碳酸氢钠或碳酸氢钾;
将中间体IMc在酸性有机试剂作用下脱除氨基保护基,制得中间体IMd;所述酸性有机试剂为乙酸乙酯-氯化氢,乙醚-氯化氢、三氟醋酸或三氟醋酸与有机溶剂的组合物;
将中间体IMd与芳香磺酰氯(RSO2Cl)在有机溶剂中、碱性作用下偶联,制得辛弗林磺酰化衍生物;所述有机溶剂为二氯甲烷、二甲基亚砜或N,N-二甲基甲酰胺,所述碱为碳酸钠、碳酸钾、碳酸氢钠或碳酸氢钾。
9.如权利要求1至权利要求4任一所述辛弗林磺酰化衍生物在抗细菌药物中的应用。
10.如权利要求1至权利要求4任一所述辛弗林磺酰化衍生物在抗柑橘溃疡病菌药物中的应用。
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Cited By (1)
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---|---|---|---|---|
CN114478418A (zh) * | 2022-01-28 | 2022-05-13 | 西南大学 | 辛弗林砜基四氮唑磺酰胺类衍生物、中间体及其用途 |
Citations (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005070940A2 (en) * | 2004-01-13 | 2005-08-04 | Cumbre Pharmaceuticals Inc. | Rifamycin derivatives effective against drug-resistant microbes |
EP1970377A1 (en) * | 2005-12-09 | 2008-09-17 | Meiji Seika Kaisha Ltd. | Lincomycin derivative and antibacterial agent containing the same as active ingredient |
EP2166015A1 (en) * | 2007-05-31 | 2010-03-24 | Meiji Seika Kaisha Ltd. | Lincosamide derivative, and antibacterial agent comprising the same as active ingredient |
CN102993170A (zh) * | 2012-11-23 | 2013-03-27 | 西南大学 | 克林沙星衍生物及其制备方法和用途 |
CN103396396A (zh) * | 2013-08-12 | 2013-11-20 | 西南大学 | 胺基烷酰克林沙星及其应用 |
CN106278912A (zh) * | 2016-08-03 | 2017-01-04 | 绵阳迪澳药业有限公司 | 利用结晶法制备高纯度辛弗林的生产工艺 |
US20170008847A1 (en) * | 2014-02-18 | 2017-01-12 | Nissan Chemical Industries, Ltd. | Alkynyl pyridine-substituted amide compound and pesticide |
CN107827815A (zh) * | 2017-11-10 | 2018-03-23 | 西南大学 | 氟喹诺酮类氨基衍生物及其防治柑橘病害的用途 |
CN109096278A (zh) * | 2018-09-26 | 2018-12-28 | 西南大学 | 氟喹诺酮-氮唑杂合衍生物、制备方法及其用途 |
CN109942546A (zh) * | 2019-04-16 | 2019-06-28 | 西南大学 | 喹诺酮嘧啶类化合物及其制备方法和应用 |
WO2019185551A1 (en) * | 2018-03-25 | 2019-10-03 | Snipr Biome Aps. | Treating & preventing microbial infections |
CN110759893A (zh) * | 2018-07-26 | 2020-02-07 | 南开大学 | Hyrtinadine生物碱衍生物及其制备和在抗植物病毒和病菌中的应用 |
CN111087390A (zh) * | 2017-11-10 | 2020-05-01 | 西南大学 | 氟喹诺酮类胺基衍生物及其用途 |
-
2020
- 2020-09-25 CN CN202011025692.7A patent/CN112110898B/zh active Active
Patent Citations (13)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2005070940A2 (en) * | 2004-01-13 | 2005-08-04 | Cumbre Pharmaceuticals Inc. | Rifamycin derivatives effective against drug-resistant microbes |
EP1970377A1 (en) * | 2005-12-09 | 2008-09-17 | Meiji Seika Kaisha Ltd. | Lincomycin derivative and antibacterial agent containing the same as active ingredient |
EP2166015A1 (en) * | 2007-05-31 | 2010-03-24 | Meiji Seika Kaisha Ltd. | Lincosamide derivative, and antibacterial agent comprising the same as active ingredient |
CN102993170A (zh) * | 2012-11-23 | 2013-03-27 | 西南大学 | 克林沙星衍生物及其制备方法和用途 |
CN103396396A (zh) * | 2013-08-12 | 2013-11-20 | 西南大学 | 胺基烷酰克林沙星及其应用 |
US20170008847A1 (en) * | 2014-02-18 | 2017-01-12 | Nissan Chemical Industries, Ltd. | Alkynyl pyridine-substituted amide compound and pesticide |
CN106278912A (zh) * | 2016-08-03 | 2017-01-04 | 绵阳迪澳药业有限公司 | 利用结晶法制备高纯度辛弗林的生产工艺 |
CN107827815A (zh) * | 2017-11-10 | 2018-03-23 | 西南大学 | 氟喹诺酮类氨基衍生物及其防治柑橘病害的用途 |
CN111087390A (zh) * | 2017-11-10 | 2020-05-01 | 西南大学 | 氟喹诺酮类胺基衍生物及其用途 |
WO2019185551A1 (en) * | 2018-03-25 | 2019-10-03 | Snipr Biome Aps. | Treating & preventing microbial infections |
CN110759893A (zh) * | 2018-07-26 | 2020-02-07 | 南开大学 | Hyrtinadine生物碱衍生物及其制备和在抗植物病毒和病菌中的应用 |
CN109096278A (zh) * | 2018-09-26 | 2018-12-28 | 西南大学 | 氟喹诺酮-氮唑杂合衍生物、制备方法及其用途 |
CN109942546A (zh) * | 2019-04-16 | 2019-06-28 | 西南大学 | 喹诺酮嘧啶类化合物及其制备方法和应用 |
Non-Patent Citations (6)
Title |
---|
BERRIN OZCELIK,等: "Cytotoxicity, antiviral and antimicrobial activities of alkaloids,flavonoids, and phenolic acids", 《PHARMACEUTICAL BIOLOGY》 * |
张文娟,等: "柑橘属中生物碱辛弗林生理活性的研究进展", 《食品工业科技》 * |
杜彩云,等主编: "《有机化学》", 31 December 2015, 武汉大学出版社 * |
童明庆,等: "克林沙星在体内外的抗菌活性", 《中国医院药学杂志》 * |
罗鹏: "以辛弗林为母核的新型化合物设计、合成及生物活性研究", 《中国优秀硕士学位论文全文数据库(电子期刊)工程科技I辑》 * |
韩海燕,等: "克林沙星酰胺衍生物的简易合成及其体内外生物活性", 《中国科学:化学》 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN114478418A (zh) * | 2022-01-28 | 2022-05-13 | 西南大学 | 辛弗林砜基四氮唑磺酰胺类衍生物、中间体及其用途 |
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