CN115466229A - 一种对酰胺基苯磺酰基类化合物及其应用 - Google Patents
一种对酰胺基苯磺酰基类化合物及其应用 Download PDFInfo
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- C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
- C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
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Abstract
本发明提供一种对酰胺基苯磺酰基类化合物及其应用，所述对酰胺基苯磺酰基类化合物具有如下式I所示结构。本发明提供的对酰胺基苯磺酰基类化合物，对PHGDH表现出优异的抑制活性，对PHGDH基因扩增或者PHGDH高表达的细胞株具有较强的增殖抑制活性；可作为PHGDH抑制剂，用于制备抗肿瘤药物。
Description
技术领域
本发明属于生物医药技术领域，具体涉及一种对酰胺基苯磺酰基类化合物及其应用。
背景技术
肿瘤的难治愈性很大程度上是由于肿瘤细胞内基因突变和代谢通路改变的复杂性，探索肿瘤细胞中普遍存在但又异于正常细胞的生物学特性，并针对该特性进行特异性干预，是提高肿瘤治疗效果的关键。早在上世纪二十年代，肿瘤细胞的代谢改变就引起了德国生理学家Otto Warburg的注意，即著名的“Warburg效应”，也称为“有氧糖酵解”。
随着现代分子生物学技术的发展，脂肪酸代谢、谷氨酰胺代谢、丝氨酸代谢、一碳单位代谢、胆碱代谢等诸多代谢通路的改变，也被概括到“Warburg效应”中来。越来越多的证据表明，肿瘤相关基因(致癌基因的激活或抑癌基因的丢失、失活)、细胞微环境(低氧状态)、转录因子、非编码RNA等，以及代谢酶自身的突变或代谢调控蛋白的活性变化，均可导致肿瘤细胞的代谢重编程，使肿瘤细胞具有特征性的代谢模式。在临床上，基于肿瘤细胞具有较高的葡萄糖摄取能力而开发的氟18-脱氧葡萄糖PET/CT(18F-FDG PET/CT)技术可以检测大多数原发和转移的上皮性肿瘤，成为高效特异的肿瘤诊断、分期和疗效监测的手段。因此，调控肿瘤代谢对于肿瘤的早期诊断、干预和治疗具有重要的意义。
肿瘤细胞的有氧糖酵解是代谢异常的重要标志之一，所产生的中间体可以被整合到各种代谢途径中以产生核苷酸、脂质、氨基酸和NADPH等，为合成生命物质提供重要原料。其中丝氨酸合成途径(Serine Synthetic Pathway，SSP)作为糖酵解途径的一个重要分支，控制着糖酵解中间体转化为丝氨酸及其下游物质的通量。在特定压力下，肿瘤细胞不得不通过这条途径为增殖提供原料和能量。该途径主要包括三个连续的酶催化反应，具体为：在辅因子NAD+的协同作用下，磷酸甘油酸脱氢酶(PHGDH)将10％左右的糖酵解中间体3-磷酸甘油酸(3-PG)氧化为3-磷酸丙酮酸(3P-pyruvate)，然后在磷酸丝氨酸转氨酶1(PSAT1)的作用下转变为3-磷酸丝氨酸(3P-serine)，最后经过磷酸丝氨酸磷酸酶(PSPH)转化为丝氨酸(Serine)。丝氨酸又可以进一步进行转化，为多种生物合成途径提供了前体原料，比如：丝氨酸可以被丝氨酸羟甲基转移酶(SHMT)转化为甘氨酸，进而有利于核苷酸和谷胱甘肽的合成；丝氨酸的积累能够变构激活丙酮酸激酶同工酶M2(PKM2)，进而加速糖酵解通量，促进肿瘤的发生和发展。
磷酸甘油酸脱氢酶(PHGDH)作为丝氨酸合成途径第一步的关键限速酶，控制着丝氨酸的从头合成通路。多项研究表明，在一些肿瘤细胞中(尤其是雌激素受体缺乏的乳腺癌细胞、非小细胞肺癌细胞和黑色素瘤细胞)，糖酵解途径的部分中间产物进入丝氨酸合成途径，PHGDH酶活性增强、基因出现扩增且蛋白存在高表达，参与调控肿瘤细胞的增殖、凋亡及侵袭等过程。沉默PHGDH基因或者抑制PHGDH酶活力后，丝氨酸从头合成途径流量减少，可以大幅度降低这些癌细胞在体内和体外的增殖、促进细胞凋亡、抑制肿瘤细胞侵袭，而对正常表达PHGDH的肿瘤细胞并不产生影响。也有研究表明，PHGDH通过控制细胞内线粒体的氧化还原稳态，在继发性(复发性或转移性)肿瘤的形成中发挥重要作用，对晚期癌症的靶向治疗具有潜在意义。此外，采用抑制剂或者siRNA干扰PHGDH可以克服厄洛替尼(erlotinib)治疗肺癌所产生的耐药，主要是由于抑制PHGDH能引起耐药细胞(PC9和HCC827)中DNA损伤标志物γH2AX显著上升，降低还原型谷胱甘肽(GSH)和氧化型谷胱甘肽(GSSG)的比值，从而揭示PHGDH是克服肺癌EGFR-TKIs耐药的潜在靶标。因此，在肿瘤代谢重新成为研究热点的今天，PHGDH是抗肿瘤研发策略中重要的新靶点，其抑制剂不仅可以通过全新的作用机制进行肿瘤的治疗，还将有可能解决当前肿瘤治疗所产生的耐药问题，具有重要的研究价值。
目前尚没有PHGDH抑制剂进入临床研究。其主要原因如下：(1)虽然上世纪八十年Snell教授等人便发现大鼠肝癌细胞中PHGDH酶活力是正常肝组织中的1.7-10.6倍，但是一直没有引起人们的重视。直到2011年多篇功能基因组学的Nature和Cell杂志将PHGDH重新拉回到人们的视野中，其小分子抑制剂于2014年被首次报道，因此该靶点的抑制剂历经时间较短，数量和结构类型均较少；(2)已报道的抑制剂体外活性较差，酶和细胞活性均处于微摩尔级别；(3)部分化合物药代动力学性质差，影响了化合物进行体内生物学研究。
因此，寻找结构新颖、活性强、药代动力学性质好的PHGDH抑制剂是急需解决的关键科学问题，具有重要的研究价值。
发明内容
针对现有技术的不足，本发明的目的在于提供一种对酰胺基苯磺酰基类化合物及其应用。本发明提供的对酰胺基苯磺酰基类化合物可用于制备PHGDH抑制剂。
为达到此发明目的，本发明采用以下技术方案：
第一方面，本发明提供一种对酰胺基苯磺酰基类化合物，所述对酰胺基苯磺酰基类化合物具有如下式I所示结构：
其中，R1选自卤素、取代或未取代的C6-C20(例如可以是C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)芳基、取代或未取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)杂芳基、取代或未取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基、取代或未取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷氧基、取代或未取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷基羧基、取代或未取代的C3-C20(例如可以是C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)环烷基、取代或未取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)杂环烷基、取代或未取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)烷基芳基、取代或未取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)烷氧基芳基中的任意一种；
R2选自取代或未取代的C6-C20(例如可以是C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)芳基、取代或未取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)杂芳基、取代或未取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基、取代或未取代的C3-C20(例如可以是C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)环烷基、取代或未取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)杂环烷基中的任意一种；
n选自0或1。
在本发明中，所述取代的取代基各自独立地选自卤素、羧基、羟基、酰胺基、未取代或Ra取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基、未取代或Ra取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷氧基、未取代或Ra取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷基羟基、未取代或Ra取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷基羰基、未取代或Ra取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷基酰胺基、未取代或Ra取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷氧基酰胺基、NRN1RN2、未取代或Ra取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷基羧基、未取代或Ra取代的C3-C20(例如可以是C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)环烷基、未取代或Ra取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)杂环烷基、未取代或Ra取代的C6-C20(例如可以是C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)芳基、未取代或Ra取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)杂芳基中的任意一种；
RN1、RN2各自独立地选自氢、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基中的任意一种；
Ra选自卤素、羟基、硝基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷氧基或C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)卤代烷基中的任意一种。
在本发明中，所述对酰胺基苯磺酰基类化合物选自如下式II-式VI所示结构中的任意一种：
其中，R11选自卤素、取代或未取代的C4-C20(例如可以是C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)含氮杂环烷基中的任意一种；
R12选自取代或未取代的C3-C20(例如可以是C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)环烷基、取代或未取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)杂环烷基、取代或未取代的C6-C20(例如可以是C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)芳基、取代或未取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)杂芳基、取代或未取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基、取代或未取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷氧基、取代或未取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷基羧基中的任意一种；
R2选自取代或未取代的C6-C20(例如可以是C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)芳基、取代或未取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)杂芳基中的任意一种。
p选自0-5的整数(例如可以是0、1、2、3、4、5)；X选自CH2、O或NH中的任意一种；L1选自单键、取代或未取代的C6-C20(例如可以是C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)芳基、取代或未取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)杂芳基中的任意一种；
q选自0-5的整数(例如可以是0、1、2、3、4、5)；Y选自CH2、羰基、O、NH、
中的任意一种；L2选自单键、取代或未取代的C6-C20(例如可以是C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)芳基、取代或未取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)杂芳基中的任意一种；
表示基团的连接位点。
在本发明中，所述R2选自取代的C2-C20(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)杂芳基中的任意一种；
所述取代的C2-C20杂芳基的取代基选自未取代或Rb取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基、未取代或Rb取代的C6-C20(例如可以是C6、C7、C8、C9、C10、C11、C12、C13、C14、C15、C16、C17、C18、C19、C20等)芳基中的任意一种；
所述Rb选自卤素、羟基、硝基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷氧基或C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)卤代烷基中的任意一种。
在本发明中，所述R2选自取代的噻唑基、取代的噻吩基、取代的三氮唑中的任意一种；
所述取代的取代基各自独立地选自未取代或Rc取代的苯基、未取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基；
所述Rc选自卤素、羟基、硝基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷氧基或C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)卤代烷基中的任意一种。
在本发明中，所述R11选自卤素、未取代或Rd取代的C4-C12(例如可以是C4、C5、C6、C7、C8、C9、C10、C11、C12等)含氮杂环烷基中的任意一种；
所述Rd选自羧基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷氧基中的任意一种。
在本发明中，所述R12选自未取代或Re取代的C2-C12(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12等)杂环烷基、未取代或Re取代的C3-C12(例如可以是C3、C4、C5、C6、C7、C8、C9、C10、C11、C12等)环烷基、未取代或Re取代的C6-C12(例如可以是C6、C7、C8、C9、C10、C11、C12等)芳基、未取代或Re取代的C2-C12(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12等)杂芳基、未取代或Re取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基、未取代或Re取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷氧基、未取代或Re取代的C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷基羧基中的任意一种；
所述Re选自羟基、羧基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷氧基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷基羟基、C2-C12(例如可以是C2、C3、C4、C5、C6、C7、C8、C9、C10、C11、C12等)杂环烷基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷基羰基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷基酰胺基、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)烷基羧基、NRN3RN4中的任意一种；
所述RN3、RN4各自独立地选自氢、C1-C10(例如可以是C1、C2、C3、C4、C5、C6、C7、C8、C9、C10等)直链或支链烷基中的任意一种。
在本发明中，所述L1、L2各自独立地选自单键、取代或未取代的C6-C12(例如可以是C6、C7、C8、C9、C10、C11、C12等)芳基、取代或未取代的C5-C12(例如可以是C5、C6、C7、C8、C9、C10、C11、C12等)杂芳基中的任意一种。
在本发明中，所述对酰胺基苯磺酰基类化合物具有如下A2、B1-B25、C1-C6、D1-D23、E1-E5、F1-F7所示结构中的任意一种：
第二方面，本发明提供一种根据第一方面所述的对酰胺基苯磺酰基类化合物在制备磷酸甘油酸脱氢酶抑制剂中的应用。
第三方面，本发明提一种根据第一方面所述的对酰胺基苯磺酰基类化合物在制备抗肿瘤药物中的应用。
本发明优选的化合物D8对PHGDH的酶抑制活性IC50为2.8±0.1μM，对多种PHGDH基因扩增或者PHGDH高表达的细胞株具有较强的增殖抑制活性，IC50值在3.5μM至21.6μM之间。MST实验表明，D8与PHGDH蛋白具有较强的结合力，Kd值为2.76μM；D8可以抑制MDA-MB-468细胞内丝氨酸的从头合成；流式细胞实验表明，化合物D8可以阻滞细胞周期G1期；分子动力学实验及氨基酸突变实验共同揭示了D8在PHGDH NAD+结合口袋的作用模式；药代动力学实验表明，化合物D8具有优秀的口服生物利用度(F％＝82.0％)；体内抗肿瘤实验表明，化合物D8在PC9移植瘤小鼠模型中具有很好的抗肿瘤活性。
相对于现有技术，本发明具有以下有益效果：
本发明提供了一种对酰胺基苯磺酰基类化合物，对PHGDH表现出优异的抑制活性，对PHGDH基因扩增或者PHGDH高表达的细胞株具有较强的增殖抑制活性；可作为PHGDH抑制剂，用于制备抗肿瘤药物。
附图说明
图1为D8和NAD+竞争测试的Lineweaver-Burk图。
图2为D8和3-PG竞争测试的Lineweaver-Burk图。
图3为D8在cis-PHGDH模型中的结合模式图。
图4为D8在trans-PHGDH模型中的结合模式图。
图5为D8抑制MDA-MB-468细胞中13C6-葡萄糖合成13C3-丝氨酸测试结果图。
图6为化合物D8对癌细胞系的抗增殖活性测试结果图。
图7为静脉注射和口服D8后小鼠血浆中药物浓度分布。
图8为肿瘤体积的变化曲线图。
图9为处理第31天肿瘤的重量。
图10为小鼠体重增长曲线图。
具体实施方式
下面通过具体实施方式来进一步说明本发明的技术方案。本领域技术人员应该明了，所述实施例仅仅是帮助理解本发明，不应视为对本发明的具体限制。
以下实施例和测试例中英文缩写的解释如下所示：
PHGDH，磷酸甘油酸脱氢酶；SSP，丝氨酸合成途径；3-PG，3-磷酸甘油酸；3-PPyr，3-磷酸羟基丙酮酸；NAD，烟酰胺腺嘌呤二核苷酸；PK，药代动力学；MST，微量热泳动；TGI，肿瘤生长抑制；TNBC，三阴性乳腺癌；AUC，曲线下的平均面积；MD，分子动力学。
(一)化合物A1，化合物B1-B25和合成
将市售4-(氯磺酰基)苯甲酸(化合物1)与4-苯基噻唑-2-胺(化合物2)在三乙胺存在下反应，生成化合物A1。在室温下使用三乙胺作为碱，用不同的胺衍生物与A1反应，生成化合物B1-B3、B5-B12、B14-B21、B24-B25和化合物3-6。化合物3-6在1M NaOH水溶液和CH3OH溶液中水解，生成B4、B13、B22和B23。对于化合物B16的合成，通过两步连续反应获得关键中间体9。首先，以四氢呋喃作为溶剂，在吡啶存在下，用市售4,6-二甲基嘧啶-2-胺(化合物7)和4-(氯磺酰基)苯甲酸甲酯(化合物8)反应，生成相应的酯；随后酯在氢氧化钠水溶液中水解生成中间体(化合物9)，中间体(化合物9)与相应的胺反应生成化合物B16。
合成反应式如下所示：
化合物A1、B1-B3、B5-B12、B14-B21、B24和B25的合成：
化合物B4、B13、B22和B23的合成：
化合物B16的合成：
反应条件：(a)Et3N,THF,reflux,4h；(b)Et3N,THF,rt,2h；(c)1M NaOH(aq.),CH3OH,rt,2h；(d)Pyridine,THF,rt,4h；(e)1M NaOH(aq.),CH3OH,rt,2h；(f)HATU,DIPEA,THF,rt,6h.
(二)化合物C1-C6和化合物D1-D23的合成
化合物C1-C6和化合物D1-D23的合成步骤同化合物B16的合成，合成反应式如下所示：
反应条件：(a)Et3N,THF,reflux,4h；(b)1M NaOH(aq.),CH3OH,rt,2h；(c)HATU,DIPEA,THF,rt,4h.
制备例1
化合物9的合成：在室温下，向4,6-二甲基嘧啶-2-胺(化合物7,123mg,1.0mmol)的THF(8mL)溶液中加入4-(氯磺酰基)苯甲酸甲酯(化合物8,235mg,1.0mmol)和吡啶(243μL，3.0mmol)，搅拌约4小时后，减压除去溶剂。将所得残余物溶解在CH3OH(3mL)和NaOH(1mol/L)水溶液(3mL)中，再搅拌2小时。然后减压除去溶剂，用HCl(1mmol/L)水溶液将残余物酸化至pH＝2或更低。溶液用EtOAc(3×20mL)萃取，合并的有机物用无水Na2SO4干燥并真空浓缩。通过硅胶色谱法(DCM/CH3OH＝10/1，v/v)纯化得到的残余物，得到化合物9，白色固体184mg，产率60％。
1H NMR(600MHz,DMSO-d6)δ8.07(m,4H),6.75(s,1H),2.25(s,6H).ESI-MS:calcdfor[M+H]+m/z 308.1,found:308.2.
制备例2
化合物11的合成步骤同化合物9，以化合物8和甲基-3-氨基苯乙酮(化合物10)制备得到。白色固体，产率72％。1H NMR(600MHz,DMSO-d6)δ10.75(s,1H),8.08(d,J＝8.4Hz,2H),7.89(d,J＝8.4Hz,2H),7.68(d,J＝7.7Hz,1H),7.65(d,J＝1.7Hz,1H),7.42(t,J＝7.9Hz,1H),7.37(dd,J＝8.1,1.2Hz,1H),2.51(s,3H).ESI-MS:calcd for[M-H]-m/z318.0,found:318.1.
实施例1
化合物A1的合成：以THF(20mL)作溶剂，将4-(氯磺酰基)苯甲酸(化合物1，1.14g，5.16mmol)、4-苯基噻唑-2-胺(化合物2，1.0g，5.67mmol)和Et3N(786μL，5.67mmol)混合，回流反应4h。减压浓缩移除溶剂后，添加EtOAc(100mL)和水(20mL)。分液，水层用EtOAc(3×20mL)萃取，合并有机层，用无水Na2SO4干燥并真空浓缩。通过硅胶柱层析(PE/EA＝10/1，v/v)纯化粗化合物，得到淡黄色固体A1(879mg，45％产率)。
1H NMR(600MHz,DMSO-d6)δ12.84(s,1H),8.07(d,J＝8.3Hz,2H),7.97(d,J＝7.3Hz,2H),7.74(d,J＝8.3Hz,2H),7.71(s,1H),7.46(t,J＝7.7Hz,2H),7.34(t,J＝7.3Hz,1H).13CNMR(150MHz,DMSO-d6)δ165.47,158.96,152.41,149.58,134.82,132.37,129.22,128.44,128.29,126.25,126.10,109.05.
实施例2
化合物B1的合成：在室温下，向吗啉(87.6μL，1.0mmol)的THF(8mL)溶液中加入化合物A1(379mg，1.0mmol)和Et3N(278μL，2.0mmol)。将混合物搅拌反应4小时并通过TLC监测。减压浓缩除去溶剂后，加入EtOAc(100mL)和水(20mL)。有机层用无水Na2SO4干燥并真空浓缩。通过硅胶柱色谱法(PE/EA＝4/1，v/v)纯化粗化合物，得到化合物B1(365mg，85％产率)，得到白色固体B1。
1H NMR(600MHz,DMSO-d6)δ13.09(s,1H),8.36(d,J＝8.4Hz,2H),7.97(d,J＝7.2Hz,2H),7.91(d,J＝8.4Hz,2H),7.76(s,1H),7.46(t,J＝7.7Hz,2H),7.35(t,J＝7.3Hz,1H),3.68–3.62(m,4H),2.97–2.91(m,4H).13C NMR(150MHz,DMSO-d6)δ169.75,163.52,157.60,148.69,137.25,135.67,133.65,128.76,128.19,127.31,127.22,125.20,108.32,64.69,45.29.HRMS(ESI):[M+H]+calcd for C20H20N3O4S2 +430.0890,found 430.0894.HPLCpurity,99％.
实施例3
化合物B2的合成步骤同化合物B1，由化合物A1和硫代吗啉制备得到。白色固体，产率90％。1H NMR(600MHz,DMSO-d6)δ13.07(s,1H),8.34(d,J＝8.4Hz,2H),7.97(d,J＝7.4Hz,2H),7.92(d,J＝8.4Hz,2H),7.76(s,1H),7.46(t,J＝7.7Hz,2H),7.35(t,J＝7.3Hz,1H),3.30-3.25(m,4H),2.71–2.67(m,4H).13C NMR(150MHz,DMSO-d6)δ164.59,158.68,149.76,140.03,136.60,134.72,129.90,129.25,128.38,127.86,126.27,109.38,48.27,26.89.HRMS(ESI):[M+Na]+calcd for C20H19N3O3S3Na+468.0481,found 468.0476.HPLCpurity,99％.
实施例4
化合物B3的合成步骤同化合物B1，由化合物A1和硫代吗啉-1,1-二氧化物制备得到，白色固体，产率87％。1H NMR(600MHz,DMSO-d6)δ13.08(s,1H),8.36(d,J＝8.3Hz,2H),8.00(d,J＝8.3Hz,2H),7.96(d,J＝7.7Hz,2H),7.76(s,1H),7.46(t,J＝7.6Hz,2H),7.35(t,J＝7.4Hz,1H),3.56-3.48(s,4H),3.33–3.28(m,4H).13C NMR(150MHz,DMSO-d6)δ164.55,158.67,149.76,139.79,137.01,134.71,130.08,129.26,128.39,127.92,126.27,109.41,50.63,45.57.HRMS(ESI):[M+H]+calcd for C20H20N3O5S3 +478.0560,found478.0557.HPLC purity,99％.
实施例5
化合物B5的合成步骤同化合物B1，由化合物A1和4-哌啶乙醇制备得到，白色固体，产率75％。1H NMR(600MHz,CDCl3)δ10.90(s,1H),7.95(d,J＝8.3Hz,2H),7.73(t,J＝8.7Hz,4H),7.35(t,J＝7.6Hz,2H),7.28(d,J＝7.4Hz,1H),7.24(s,1H),3.78(d,J＝11.7Hz,2H),3.67(t,J＝6.3Hz,2H),2.27-2.20(m,2H),1.76(d,J＝12.7Hz,2H),1.50(q,J＝6.4Hz,2H),1.44–1.37(m,1H),1.36–1.28(m,2H).13C NMR(150MHz,CDCl3)δ163.67,158.46,150.33,140.26,135.64,133.87,128.85,128.31,128.09,128.02,126.06,108.55,60.02,46.38,38.61,31.66,31.40.HRMS(ESI):[M+H]+calcd for C23H26N3O4S2 +472.1359,found472.1360.HPLC purity,99％.
实施例6
化合物B6的合成步骤同化合物B1，由化合物A1和环己胺制备得到，白色固体，产率83％。1H NMR(600MHz,DMSO-d6)δ13.02(s,1H),8.28(d,J＝8.4Hz,2H),7.99–7.95(m,4H),7.88(d,J＝7.4Hz,1H),7.74(s,1H),7.46(t,J＝7.7Hz,2H),7.35(t,J＝7.3Hz,1H),3.07–2.95(m,1H),1.58(d,J＝8.2Hz,4H),1.45(d,J＝12.1Hz,1H),1.20–1.10(m,4H),1.07-0.98(m,1H).13C NMR(150MHz,DMSO-d6)δ164.78,158.71,149.72,146.14,135.67,134.74,129.62,129.25,128.36,126.82,126.25,109.31,52.67,33.73,25.28,24.81.HRMS(ESI):[M+H]+calcd forC22H24N3O3S2 +442.1254,found 442.1261.HPLC purity,97％.
实施例7
化合物B7的合成步骤同化合物B1，由化合物A1和4-氨基四氢吡喃制备得到，白色固体，产率30％，1H NMR(600MHz,DMSO-d6)δ13.01(s,1H),8.28(d,J＝6.7Hz,2H),8.08–7.91(m,5H),7.75(s,1H),7.45(t,J＝5.6Hz,2H),7.35(t,J＝8.2Hz,1H),3.72(d,J＝9.2Hz,2H),3.29–3.18(m,3H),1.53(d,J＝10.4Hz,2H),1.37(d,J＝9.3Hz,2H).13C NMR(150MHz,DMSO-d6)δ164.75,158.70,149.74,145.95,135.82,134.74,129.69,129.26,128.38,126.87,126.26,109.34,65.99,49.86,33.80.HRMS(ESI):[M+H]+calcd for C21H22N3O4S2 +444.1046,found 444.1049.HPLC purity,96％.
实施例8
化合物B8的合成步骤同化合物B1，由化合物A1和1-甲基哌啶-4-胺制备得到，白色固体，产率75％。1H NMR(600MHz,DMSO-d6)δ8.29(d,J＝8.4Hz,2H),8.01(d,J＝7.0Hz,1H),7.97(dd,J＝7.6,6.3Hz,4H),7.74(s,1H),7.46(t,J＝7.7Hz,2H),7.35(t,J＝7.4Hz,1H),3.09–3.01(m,1H),2.74(d,J＝10.2Hz,2H),2.20(s,3H),2.14–2.02(m,2H),1.59(d,J＝10.8Hz,2H),1.51–1.41(m,2H).13C NMR(150MHz,DMSO-d6)δ164.89,159.09,149.63,145.76,136.03,134.77,129.65,129.24,128.33,126.85,126.25,109.22,54.13,46.03,32.63.HRMS(ESI):[M+H]+calcd for C22H25N4O3S2 +457.1363,found 457.1360.HPLCpurity,98％.
实施例9
化合物B9的合成步骤同化合物B1，由化合物A1和1-乙酰哌啶-4-胺制备得到，白色固体，产率73％。1H NMR(600MHz,DMSO-d6)δ13.02(s,1H),8.29(d,J＝8.4Hz,2H),8.04(d,J＝7.3Hz,1H),7.98(dd,J＝13.8,7.9Hz,4H),7.75(s,1H),7.46(t,J＝7.7Hz,2H),7.35(t,J＝7.3Hz,1H),4.07(d,J＝13.2Hz,1H),3.65(d,J＝13.9Hz,1H),3.32–3.25(m,1H),3.08–2.99(m,1H),2.71–2.62(m,1H),1.94(s,3H),1.67–1.53(m,2H),1.29(td,J＝14.5,4.0Hz,1H),1.19(td,J＝14.8,4.2Hz,1H).13C NMR(150MHz,DMSO-d6)δ168.37,164.74,158.73,149.73,145.80,135.85,134.74,129.71,129.25,128.37,126.87,126.26,109.33,50.58,44.51,40.52,33.36,32.51,21.65.HRMS(ESI):[M+H]+calcd for C23H25N4O4S2 +485.1312,found 485.1314.HPLC purity,99％.
实施例10
化合物B10的合成步骤同化合物B1，由化合物A1和4-甲基-1-哌嗪乙胺制备得到，白色固体，产率90％。1H NMR(600MHz,CDCl3)δ7.98(d,J＝7.4Hz,2H),7.89(d,J＝7.6Hz,2H),7.72(d,J＝7.0Hz,2H),7.35(t,J＝6.8Hz,2H),7.29(d,J＝7.0Hz,1H),7.23(s,1H),3.51–3.49(m,1H),3.02–2.97(m,2H),2.45–2.25(m,12H).13C NMR(150MHz,DMSO-d6)δ163.74,157.91,148.57,143.41,134.87,133.67,128.51,128.17,127.26,125.99,125.18,108.19,56.22,53.82,51.73,44.92,39.55.HRMS(ESI):[M+H]+calcd for C23H28N5O3S2 +486.1628,found486.1642.HPLC purity,98％.
实施例11
化合物B11的制备步骤同化合物B1，由化合物A1和4-(2-氨基乙基)吗啉制备得到，白色固体，产率79％。1H NMR(600MHz,CDCl3)δ10.57(s,1H),8.01(d,J＝7.8Hz,2H),7.93(d,J＝7.6Hz,2H),7.75(d,J＝7.1Hz,2H),7.37(t,J＝7.0Hz,2H),7.30(t,J＝6.9Hz,1H),7.24(s,1H),3.65–3.58(m,4H),3.04(t,J＝5.2Hz,2H),2.42(t,J＝5.2Hz,2H),2.33–2.25(m,4H).13CNMR(150MHz,CDCl3)δ163.32,158.07,150.34,143.67,135.70,133.92,128.85,128.31,128.21,127.57,126.07,108.55,66.75,56.26,52.96,38.94.HRMS(ESI):[M+H]+calcd for C22H25N4O4S2 +473.1312,found 473.1314.HPLC purity,99％.
实施例12
化合物B12的合成步骤同化合物B1，由化合物A1和2-甲氧基乙胺制备得到，白色固体，产率90％。1H NMR(600MHz,CDCl3)δ11.22(s,1H),7.91(d,J＝7.4Hz,2H),7.81(d,J＝7.5Hz,2H),7.68(d,J＝6.9Hz,2H),7.31(t,J＝6.8Hz,2H),7.26(d,J＝7.8Hz,2H),5.17(s,1H),3.42–3.37(m,2H),3.25(s,3H),3.15–3.10(m,2H).13C NMR(150MHz,CDCl3)δ163.77,158.75,150.32,143.66,135.63,133.85,128.81,128.29,128.26,127.38,126.10,108.59,70.41,58.81,42.97.HRMS(ESI):[M+H]+calcd for C19H20N3O4S2 +418.0890,found418.0893.HPLCpurity,99％.
实施例13
化合物B14的合成步骤同化合物B1，由化合物A1和N,N-二甲基乙二胺制备得到，白色固体，产率79％。1H NMR(600MHz,DMSO-d6)δ12.95(s,1H),8.29(d,J＝8.4Hz,2H),7.96(dd,J＝8.1,2.5Hz,4H),7.74(s,1H),7.46(t,J＝7.7Hz,2H),7.35(t,J＝7.3Hz,1H),2.90(t,J＝6.8Hz,2H),2.27(t,J＝6.7Hz,2H),2.07(s,6H).13C NMR(150MHz,DMSO-d6)δ163.75,157.88,148.57,143.41,134.87,133.68,128.50,128.16,127.26,125.98,125.18,108.19,57.49,44.37,40.05.HRMS(ESI):[M+H]+calcd for C20H23N4O3S2 +431.1206,found431.1212.HPLC purity,98％.
实施例14
化合物B15的合成步骤同化合物B1，由化合物A1和苯胺制备得到，白色固体，产率80％。1H NMR(600MHz,DMSO-d6)δ12.98(s,1H),10.48(s,1H),8.22(d,J＝8.4Hz,2H),7.95(d,J＝7.4Hz,2H),7.90(d,J＝8.5Hz,2H),7.74(s,1H),7.45(t,J＝7.7Hz,2H),7.34(t,J＝7.3Hz,1H),7.26(t,J＝7.9Hz,2H),7.12(d,J＝7.7Hz,2H),7.06(t,J＝7.4Hz,1H).13C NMR(150MHz,DMSO-d6)δ164.61,158.64,149.71,143.17,137.77,136.40,134.71,129.76,129.68,129.24,128.36,127.29,126.24,124.91,120.80,109.33.HRMS(ESI):[M+H]+calcdfor C22H18N3O3S2 +436.0784,found 436.0792.HPLC purity,97％.
实施例15
化合物B17的合成步骤同化合物B1，由化合物A1和2H-吡唑-3-基胺制备得到，白色固体，产率73％。1H NMR(600MHz,DMSO-d6)δ13.05(s,1H),8.28(d,J＝8.4Hz,2H),8.08(d,J＝2.9Hz,1H),7.98(d,J＝8.4Hz,2H),7.95(d,J＝7.6Hz,2H),7.75(s,1H),7.45(t,J＝7.6Hz,2H),7.35(t,J＝7.3Hz,1H),5.92(d,J＝2.9Hz,1H).13C NMR(150MHz,DMSO-d6)δ164.40,161.53,158.58,149.75,140.21,137.55,135.29,134.69,129.86,129.25,128.38,127.89,126.25,109.42,102.94.HRMS(ESI):[M+H]+calcd for C19H16N5O3S2 +426.0689,found 426.0698.HPLCpurity,97％.
实施例16
化合物B18的合成步骤同化合物B1，由化合物A1和3-氨基苯甲醇制备得到，白色固体，产率85％。1H NMR(600MHz,DMSO-d6)δ12.99(s,1H),10.47(s,1H),8.23(d,J＝8.4Hz,2H),7.95(d,J＝7.5Hz,2H),7.92(d,J＝8.4Hz,2H),7.73(s,1H),7.45(t,J＝7.7Hz,2H),7.34(t,J＝7.3Hz,1H),7.20(t,J＝7.8Hz,1H),7.13(s,1H),7.03–6.97(m,2H),5.22(t,J＝5.7Hz,1H),4.41(d,J＝5.6Hz,2H).13C NMR(150MHz,DMSO-d6)δ164.64,158.64,149.71,144.41,143.30,137.70,136.36,134.70,129.67,129.38,129.24,128.36,127.28,126.25,122.80,118.81,118.61,109.32,62.94.HRMS(ESI):[M+H]+calcd for C23H20N3O4S2 +466.0890,found 466.0890.HPLCpurity,97％.
实施例17
化合物B19的合成步骤同化合物B1，由化合物A1和2-(3-氨基苯基)乙醇制备得到，白色固体，产率45％。1H NMR(600MHz,DMSO-d6)δ12.99(s,1H),10.43(s,1H),8.22(d,J＝8.4Hz,2H),7.95(d,J＝7.5Hz,2H),7.91(d,J＝8.4Hz,2H),7.73(s,1H),7.45(t,J＝7.7Hz,2H),7.35(t,J＝7.3Hz,1H),7.15(t,J＝7.8Hz,1H),6.99(s,1H),6.96-6.90(m,2H),4.67(t,J＝5.2Hz,1H),3.53-3.49(m,2H),2.62(t,J＝7.0Hz,2H).13C NMR(150MHz,DMSO-d6)δ164.63,158.63,149.71,143.23,141.27,137.59,136.33,134.69,129.63,129.46,129.25,128.38,127.33,126.24,125.50,121.21,118.24,109.32,62.39,39.34.HRMS(ESI):[M+H]+calcd for C24H22N3O4S2 +480.1046,found 480.1054.HPLC purity,95％.
实施例18
化合物B20的合成步骤同化合物B1，由化合物A1和3-(4-吗啉基)苯胺制备得到，白色固体，产率70％。1H NMR(600MHz,DMSO-d6)δ12.97(s,1H),10.33(s,1H),8.22(d,J＝8.5Hz,2H),7.95(d,J＝7.2Hz,2H),7.91(d,J＝8.5Hz,2H),7.73(s,1H),7.45(t,J＝7.7Hz,2H),7.34(t,J＝7.4Hz,1H),7.07(t,J＝8.1Hz,1H),6.66(d,J＝1.9Hz,1H),6.64(dd,J＝8.3,2.0Hz,1H),6.56(dd,J＝7.9,1.2Hz,1H),3.72–3.68(m,4H),3.02–2.98(m,4H).13C NMR(150MHz,DMSO-d6)δ151.09,148.62,142.19,137.50,135.29,133.63,129.05,128.57,128.16,127.28,126.28,125.16,110.62,110.16,108.25,105.97,65.34,47.48.HRMS(ESI):[M+H]+calcd forC26H25N4O4S2 +521.1312,found 521.1319.HPLC purity,97％.
实施例19
化合物B21的合成步骤同化合物B1，由化合物A1和3-(4-甲基哌嗪-1-基)苯胺制备得到，白色固体，产率85％。1H NMR(600MHz,DMSO-d6)δ12.95(s,1H),10.32(s,1H),8.23(d,J＝8.4Hz,2H),7.95(d,J＝7.5Hz,2H),7.91(d,J＝8.4Hz,2H),7.73(s,1H),7.45(t,J＝7.7Hz,2H),7.34(t,J＝7.3Hz,1H),7.05(t,J＝8.1Hz,1H),6.65(s,1H),6.63(d,J＝8.3Hz,1H),6.53(d,J＝7.8Hz,1H),3.07–2.99(m,4H),2.44–2.38(m,4H),2.20(s,3H).13C NMR(150MHz,DMSO-d6)δ163.61,157.73,150.96,148.56,142.22,137.47,135.35,133.64,128.99,128.54,128.15,127.26,126.27,125.17,110.90,109.91,108.20,106.37,53.75,47.11,45.02.HRMS(ESI):[M+H]+calcd for C27H28N5O3S2 +534.1628,found 534.1635.HPLCpurity,99％.
实施例20
化合物B24的合成步骤同化合物B1，由化合物A1和3-氨基苯乙酮制备得到，白色固体，产率65％。1H NMR(600MHz,DMSO-d6)δ12.99(s,1H),10.75(s,1H),8.23(d,J＝8.4Hz,2H),7.94(t,J＝9.1Hz,4H),7.73(s,1H),7.70–7.66(m,2H),7.48-7.42(m,3H),7.39(d,J＝8.3Hz,1H),7.34(t,J＝7.3Hz,1H),2.51(s,3H).13C NMR(150MHz,DMSO-d6)δ197.74,164.56,158.60,149.72,142.90,138.31,138.17,136.57,134.71,130.33,129.80,129.24,128.37,127.30,126.24,125.11,124.93,119.34,109.35,27.19.HRMS(ESI):[M+H]+calcdfor C24H20N3O4S2 +478.0890,found 478.0896.HPLC purity,98％.
实施例21
化合物B25的合成步骤同化合物B1的合成，由化合物A1和3-氨基-N-甲基苯甲酰胺制备得到，白色固体，产率80％。1H NMR(600MHz,DMSO-d6)δ12.98(s,1H),10.64(s,1H),8.44–8.40(m,1H),8.23(d,J＝8.4Hz,2H),7.95(d,J＝7.5Hz,2H),7.91(d,J＝8.4Hz,2H),7.73(s,1H),7.63(s,1H),7.50(d,J＝7.7Hz,1H),7.45(t,J＝7.7Hz,2H),7.34(t,J＝7.9Hz,2H),7.28–7.24(m,1H),2.74(d,J＝4.5Hz,3H).13C NMR(150MHz,DMSO-d6)δ166.43,164.57,158.61,149.71,143.03,138.00,136.48,136.21,134.71,129.74,129.71,129.24,128.36,127.27,126.24,123.09,120.01,109.34,26.74.HRMS(ESI):[M+H]+calcd forC24H21N4O4S2 +493.0999,found493.1008.HPLC purity,97％.
实施例22
化合物B4的合成：在室温下，向甲基哌啶-4-羧酸盐(135μL，1.0mmol)的THF(8mL)溶液中加入化合物A1(379mg，1.0mmol)和Et3N(278μL，2.0mmol)，搅拌约4小时后，减压除去溶剂。将所得残余物溶解在CH3OH(3mL)和NaOH(1mol/L)水溶液(3mL)中并再搅拌2小时。然后减压除去溶剂，用HCl(1mol/L)水溶液将残余物酸化至pH＝2或更低。溶液用EtOAc(3×20mL)萃取，合并的有机物用无水Na2SO4干燥并真空浓缩。通过硅胶色谱法(DCM/CH3OH＝10/1，v/v)纯化得到的残余物，得到呈白色固体状的所需化合物B4(396mg，产率84.0％)。
1H NMR(600MHz,DMSO-d6)δ13.08(s,1H),12.37(s,1H),8.35(d,J＝8.4Hz,2H),7.97(d,J＝7.5Hz,2H),7.91(d,J＝8.3Hz,2H),7.76(s,1H),7.46(t,J＝7.7Hz,2H),7.35(t,J＝7.3Hz,1H),3.56-3.50(m,2H),2.51-2.47(m,2H),2.36–2.27(m,1H),1.94–1.85(m,2H),1.61-1.52(m,2H).13C NMR(150MHz,DMSO-d6)δ175.63,164.63,139.44,136.47,134.69,129.79,129.26,128.38,128.07,126.27,109.38,45.66,39.17,27.63.HRMS(ESI):[M+H]+calcd forC22H22N3O5S2 +472.0995,found 472.0995.HPLC purity,97％.
实施例23
化合物B13的合成步骤同化合物B4的合成，以化合物A1和甘氨酸甲酯盐酸盐制备得到。白色固体，产率75％。1H NMR(600MHz,DMSO-d6)δ13.03(s,1H),12.76(s,1H),8.28(d,J＝8.4Hz,3H),7.96(t,J＝8.7Hz,4H),7.75(s,1H),7.46(t,J＝7.7Hz,2H),7.35(t,J＝7.3Hz,1H),3.66(d,J＝3.6Hz,2H).13C NMR(150MHz,DMSO-d6)δ170.68,164.72,158.80,149.67,144.59,135.79,134.72,129.48,129.25,128.37,127.12,126.26,109.32,44.39.HRMS(ESI):[M+H]+calcd for C18H16N3O5S2 +418.0526,found 418.0522.HPLCpurity,98％.
实施例24
化合物B22的合成步骤同化合物B4的合成，以化合物A1和3-氨基苯甲酸甲酯制备得到。白色固体，产率95％。1H NMR(600MHz,DMSO-d6)δ13.11(s,1H),12.99(s,1H),10.75(s,1H),8.23(d,J＝8.4Hz,2H),7.95(d,J＝7.4Hz,2H),7.91(d,J＝8.4Hz,2H),7.73(d,J＝8.6Hz,2H),7.63(d,J＝6.6Hz,1H),7.45(t,J＝7.7Hz,2H),7.42–7.38(m,2H),7.35(t,J＝7.3Hz,1H).13CNMR(150MHz,DMSO-d6)δ167.14,142.89,138.14,132.33,130.16,129.79,129.25,128.37,127.25,126.24,125.66,124.75,121.21,109.34.HRMS(ESI):[M+H]+calcdfor C23H18N3O5S2 +480.0682,found 480.0689.HPLC purity,97％.
实施例25
化合物B23的合成步骤同化合物B4的合成，以化合物A1和3-氨基苯乙酸甲酯制备得到。白色固体，产率78％。1H NMR(600MHz,DMSO-d6)δ12.97(s,1H),10.51(s,1H),8.21(d,J＝8.4Hz,2H),7.95(d,J＝7.4Hz,2H),7.92(d,J＝8.4Hz,2H),7.73(s,1H),7.45(t,J＝7.7Hz,2H),7.35(t,J＝7.3Hz,1H),7.19(t,J＝7.9Hz,1H),7.08(s,1H),7.00(d,J＝8.2Hz,1H),6.95(d,J＝7.6Hz,1H),3.49(s,2H).13C NMR(150MHz,DMSO-d6)δ172.85,164.67,158.69,149.64,143.15,137.72,136.65,134.65,129.65,129.60,129.26,128.38,127.32,126.24,125.95,121.47,118.81,109.31,41.06.HRMS(ESI):[M+H]+calcd for C24H20N3O5S2 +494.0839,found 494.0844.HPLC purity,97％.
实施例26
化合物B16的合成：在室温下向HATU(684mg,1.8mmol)、DIPEA(523μL,3.0mmol)的THF(1mL)溶液中加入化合物9(368mg,1.2mmol)。然后加入2-氨基-4-苯基噻唑(176mg，1.0mmol)，搅拌约6小时。除去溶剂，残余物用乙酸乙酯溶解，用水和饱和氯化铵水溶液洗涤。将有机相用无水Na2SO4干燥，减压浓缩，并通过快速柱色谱法纯化以获得呈白色固体状的化合物B16(158mg，产率34％)。
1H NMR(600MHz,CDCl3)δ8.22(d,J＝7.9Hz,2H),7.99(d,J＝7.9Hz,2H),7.75(d,J＝7.1Hz,2H),7.36(t,J＝7.0Hz,2H),7.29(d,J＝7.0Hz,1H),7.21(s,1H),6.65(s,1H),2.34(s,6H).13C NMR(150MHz,CDCl3)δ168.36,163.44,158.00,155.67,150.33,143.45,135.98,133.99,129.44,128.81,128.25,127.45,126.08,115.19,108.48,23.57.HRMS(ESI):[M+H]+calcd for C22H20N5O3S2 +466.1002,found 466.1004.HPLC purity,98％.
实施例27
化合物C1的合成步骤同化合物B16，以化合物11和4-苯基噻吩-2-胺制备得到，白色固体，产率52％。1H NMR(600MHz,DMSO-d6)δ11.85(s,1H),10.73(s,1H),8.12(d,J＝8.4Hz,2H),7.95(d,J＝8.4Hz,2H),7.68(d,J＝7.1Hz,2H),7.63(d,J＝7.6Hz,2H),7.45–7.37(m,5H),7.29(t,J＝7.3Hz,1H),7.25(d,J＝1.4Hz,1H),2.51(s,3H).13C NMR(150MHz,DMSO-d6)δ197.75,162.68,142.37,140.85,138.35,138.15,137.55,135.72,130.31,129.41,129.21,127.57,127.45,126.16,125.10,124.92,119.31,113.65,111.50,27.19.HRMS(ESI):[M-H]-calcd for C25H19N2O4S2 -475.0792,found 475.0797.HPLCpurity,95％.
实施例28
化合物C2的合成步骤同化合物B16，以化合物11和5-甲基-4-苯基噻唑-2-胺制备得到，白色固体，产率40％。1H NMR(600MHz,DMSO-d6)δ12.85(s,1H),10.75(s,1H),8.21(d,J＝8.5Hz,2H),7.92(d,J＝8.5Hz,2H),7.70–7.66(m,4H),7.47(t,J＝7.7Hz,2H),7.43(t,J＝7.8Hz,1H),7.41–7.34(m,2H),2.51(s,3H),2.50(s,3H).13C NMR(150MHz,DMSO-d6)δ196.66,141.72,137.23,137.08,129.25,128.66,127.79,127.47,126.78,126.22,124.03,123.84,118.25,26.11,11.23.HRMS(ESI):[M-H]-calcd for C25H20N3O4S-490.0901,found490.0910.HPLCpurity,96％.
实施例29
化合物C3的合成步骤同化合物B16，以化合物11和4-苯基-1H-咪唑-2-胺制备得到，白色固体，产率36％。1H NMR(600MHz,DMSO-d6)δ12.06(s,1H),11.95(s,1H),10.72(s,1H),8.18(d,J＝8.3Hz,2H),7.90(d,J＝8.4Hz,2H),7.76(d,J＝7.6Hz,2H),7.67(dd,J＝6.5,5.1Hz,2H),7.43(t,J＝7.8Hz,1H),7.39(d,J＝8.3Hz,2H),7.36(t,J＝7.7Hz,2H),7.20(t,J＝7.3Hz,1H),2.51(s,3H).13C NMR(150MHz,DMSO-d6)δ197.75,138.38,138.15,130.30,129.48,129.02,127.21,125.05,124.89,124.55,119.29,27.19.HRMS(ESI):[M-H]-calcd forC24H19N4O4S-459.1132,found 459.1135.HPLC purity,97％.
实施例30
化合物C4的合成步骤同化合物B16，以化合物11和1-甲基-3-苯基-1H-吡唑-5-胺制备得到，白色固体，产率64％。1H NMR(600MHz,DMSO-d6)δ10.76(s,1H),10.64(s,1H),8.12(d,J＝8.3Hz,2H),7.95(d,J＝8.4Hz,2H),7.78(d,J＝7.3Hz,2H),7.71–7.65(m,2H),7.46–7.38(m,4H),7.30(t,J＝7.4Hz,1H),6.72(s,1H),3.75(s,3H),2.52(s,3H).13C NMR(150MHz,DMSO-d6)δ197.79,164.97,148.69,142.64,138.42,138.16,137.84,137.59,133.75,130.33,129.51,129.12,127.96,127.37,125.24,125.05,124.81,119.19,98.07,36.41,27.21.HRMS(ESI):[M-H]-calcd for C25H21N4O4S-473.1289,found 473.1291.HPLCpurity,96％.
实施例31
化合物C5的合成步骤同化合物B16，以化合物11和5-苯基噻唑-2-胺制备得到，白色固体，产率47％。1H NMR(600MHz,DMSO-d6)δ12.93(s,1H),10.75(s,1H),8.21(d,J＝8.4Hz,2H),7.99(s,1H),7.93(d,J＝8.5Hz,2H),7.69–7.65(m,4H),7.44(t,J＝7.8Hz,3H),7.41–7.38(m,1H),7.33(t,J＝7.4Hz,1H),2.51(s,3H).13C NMR(150MHz,DMSO-d6)δ197.75,142.84,138.31,138.16,131.85,130.33,129.79,129.71,129.40,128.20,127.32,126.24,125.11,124.91,119.31,27.20.HRMS(ESI):[M-H]-calcd for C24H18N3O4S2 -476.0744,found476.0764.HPLCpurity,95％.
实施例32
化合物C6的合成步骤同化合物B16，以化合物11和5-苯基-4H-1,2,4-三唑-3-胺制备得到，白色固体，产率45％。1H NMR(600MHz,DMSO-d6)δ10.79(s,1H),8.25–8.22(m,2H),7.97–7.94(m,2H),7.92–7.89(m,3H),7.70(dd,J＝7.1,1.4Hz,2H),7.49–7.40(m,5H),2.52(s,3H).13C NMR(150MHz,DMSO-d6)δ197.72,166.85,160.40,159.29,142.85,138.32,138.17,136.76,132.02,130.70,130.37,130.33,129.16,126.97,126.74,125.13,125.01,119.40,27.19.HRMS(ESI):[M-H]-calcd for C23H18N5O4S-460.1085,found 460.1088.HPLCpurity,96％.
实施例33
化合物D1的合成步骤同化合物B16，以化合物11和4-(2-氟苯基)噻唑-2-胺制备得到，白色固体，产率57％。1H NMR(600MHz,DMSO-d6)δ13.03(s,1H),10.76(s,1H),8.23(d,J＝8.4Hz,2H),8.09(dd,J＝8.1,6.8Hz,1H),7.94(d,J＝8.4Hz,2H),7.70–7.66(m,2H),7.63(d,J＝2.1Hz,1H),7.44(t,J＝7.8Hz,1H),7.42–7.36(m,2H),7.36–7.30(m,2H),2.52(s,3H).13CNMR(150MHz,DMSO-d6)δ197.74,164.66,160.85,159.20,158.14,143.36,142.92,138.31,138.16,136.51,130.33,130.05,129.83,129.78,127.31,125.26,125.11,124.92,122.30,119.32,116.76,116.61,113.76,27.20.HRMS(ESI):[M-H]-calcd forC24H17FN3O4S2 -494.0650,found494.0656.HPLC purity,97％.
实施例34
化合物D2的合成步骤同化合物B16，以化合物11和4-(2-氯苯基)噻唑-2-胺制备得到，白色固体，产率38％。1H NMR(600MHz,Acetone-d6)δ8.29(d,J＝8.2Hz,2H),8.01(d,J＝8.4Hz,2H),7.92(d,J＝7.7Hz,1H),7.84(s,1H),7.74(d,J＝7.7Hz,1H),7.71(s,1H),7.54–7.49(m,2H),7.47–7.40(m,2H),7.36(t,J＝7.6Hz,1H),2.53(s,3H).13C NMR(150MHz,Acetone-d6)δ198.20,165.64,158.87,148.25,144.88,140.12,139.73,138.25,135.13,133.31,133.15,132.22,131.48,130.90,130.70,129.21,128.84,126.76,126.55,121.76,115.12,27.62.HRMS(ESI):[M-H]-calcd for C24H19ClN3O4S2 -512.0511,found512.0506.HPLC purity,99％.
实施例35
化合物D3的合成步骤同化合物B16，以化合物11和4-(邻甲苯基)噻唑-2-胺制备得到，白色固体，产率59％。1H NMR(600MHz,DMSO-d6)δ12.93(s,1H),10.75(s,1H),8.21(d,J＝8.5Hz,2H),7.92(d,J＝8.5Hz,2H),7.70–7.65(m,2H),7.64–7.58(m,1H),7.43(t,J＝7.8Hz,1H),7.41-7.35(m,2H),7.31–7.24(m,3H),2.51(s,3H),2.44(s,3H).13C NMR(150MHz,DMSO-d6)δ197.74,142.85,138.33,138.16,135.92,131.27,130.33,129.88,129.78,128.30,127.31,126.30,125.11,124.91,119.32,112.11,27.19,21.44.HRMS(ESI):[M-H]-calcd forC25H20N3O4S2 -490.0901,found 490.0916.HPLC purity,100％.
实施例36
化合物D4的合成步骤同化合物B16，以化合物11和4-(2-(三氟甲基)苯基)噻唑-2-胺制备得到，白色固体，产率49％。1H NMR(600MHz,DMSO-d6)δ12.99(s,1H),10.75(s,1H),8.21(d,J＝8.5Hz,2H),7.92(d,J＝8.5Hz,2H),7.85(d,J＝7.9Hz,1H),7.75(t,J＝7.5Hz,1H),7.70–7.62(m,4H),7.43(t,J＝7.8Hz,1H),7.39(dd,J＝8.1,1.0Hz,1H),7.35(s,1H),2.51(s,3H).13C NMR(150MHz,DMSO-d6)δ197.73,164.67,157.98,147.17,142.90,138.31,138.16,136.53,134.86,132.77,132.61,130.33,129.80,129.22,127.58,127.39,127.29,126.75,125.45,125.10,124.90,123.63,119.31,113.28,27.19.HRMS(ESI):[M-H]-calcdfor C25H17F3N3O4S2 -544.0618,found 544.0628.HPLC purity,99％.
实施例37
化合物D5的合成步骤同化合物B16，以化合物11和4-(2-甲氧基苯基)噻唑-2-胺制备得到，白色固体，产率65％。1H NMR(600MHz,DMSO-d6)δ12.91(s,1H),10.76(s,1H),8.23(d,J＝8.3Hz,2H),8.13(d,J＝7.3Hz,1H),7.94(d,J＝8.3Hz,2H),7.75(s,1H),7.69(d,J＝6.9Hz,2H),7.44(t,J＝8.0Hz,1H),7.40(d,J＝8.0Hz,1H),7.33(t,J＝7.6Hz,1H),7.14(d,J＝8.3Hz,1H),7.05(t,J＝7.5Hz,1H),3.93(s,3H),2.52(s,3H).13C NMR(150MHz,DMSO-d6)δ197.73,157.11,142.84,138.32,138.17,130.33,129.78,129.45,129.39,127.29,125.10,124.92,120.91,119.33,112.95,112.16,55.94,27.19.HRMS(ESI):[M-H]-calcdfor C25H20N3O5S2 -506.0850,found 506.0864.HPLC purity,99％.
实施例38
化合物D6的合成步骤同化合物B16，以化合物11和2-(2-氨基噻唑-4-基)苯酚制备得到，白色固体，产率18％。1H NMR(600MHz,Acetone-d6)δ8.27(d,J＝8.4Hz,2H),8.02(d,J＝8.3Hz,2H),7.86–7.80(m,2H),7.75(d,J＝7.6Hz,1H),7.68(s,1H),7.50(dd,J＝8.1,1.1Hz,1H),7.45(t,J＝7.9Hz,1H),7.21(t,J＝7.2Hz,1H),6.92–6.86(m,2H),2.53(s,3H).13C NMR(150MHz,Acetone-d6)δ198.20,157.68,149.85,145.06,140.12,139.69,131.49,130.64,129.34,128.27,126.81,126.60,121.78,121.05,119.20,109.09,27.62.HRMS(ESI):[M+H]+calcd forC24H21N4O4S2 +493.0999,found 493.1008.HRMS(ESI):[M+H]+calcdfor C24H20N3O5S2 +494.0839,found 494.0848.HPLC purity,99％.
实施例39
化合物D7的合成步骤同化合物B16，以化合物11和4-(2-硝基苯基)噻唑-2-胺制备得到，白色固体，产率31％。1H NMR(600MHz,Acetone-d6)δ8.26(d,J＝8.4Hz,2H),7.99(d,J＝8.5Hz,2H),7.87–7.81(m,3H),7.76–7.71(m,2H),7.65–7.60(m,1H),7.52(s,1H),7.51–7.48(m,1H),7.44(t,J＝7.9Hz,1H),2.52(s,3H).13C NMR(150MHz,Acetone-d6)δ198.19,165.76,159.82,151.11,147.79,144.88,140.11,139.71,138.14,133.94,132.64,131.46,130.90,130.70,130.51,129.19,126.81,126.55,125.68,121.83,113.93,27.61.HRMS(ESI):[M+Na]+calcd forC24H18N4NaO6S2 +545.0560,found 545.0573.HPLC purity,97％.
实施例40
化合物D8的合成步骤同化合物B16，以化合物11和4-(3-氟苯基)噻唑-2-胺制备得到，白色固体，产率63％。1H NMR(600MHz,DMSO-d6)δ12.98(s,1H),10.73(s,1H),8.23(d,J＝8.5Hz,2H),7.93(d,J＝8.5Hz,2H),7.86(s,1H),7.80(d,J＝7.8Hz,1H),7.76–7.73(m,1H),7.70–7.67(m,2H),7.52–7.47(m,1H),7.44(t,J＝7.8Hz,1H),7.41–7.38(m,1H),7.18(td,J＝8.5,2.4Hz,1H),3.34(s,3H).13C NMR(150MHz,DMSO-d6)δ197.74,164.65,163.87,162.26,158.75,148.37,142.93,138.30,138.16,136.48,131.34,130.33,129.82,127.31,125.11,124.93,122.27,119.32,114.97,112.70,110.73,27.20.HRMS(ESI):[M+H]+calcdfor C24H19FN3O4S2 +496.0796,found 496.0798.HPLC purity,100％.
实施例41
化合物D9的合成步骤同化合物B16，以化合物11和4-(3-氯苯基)噻唑-2-胺制备得到，白色固体，产率29％。1H NMR(600MHz,Acetone-d6)δ8.30(d,J＝8.4Hz,2H),8.02(d,J＝8.4Hz,2H),7.97(s,1H),7.88(d,J＝7.5Hz,1H),7.85(s,1H),7.74(d,J＝7.6Hz,1H),7.69(d,J＝2.0Hz,1H),7.53–7.49(m,1H),7.47–7.41(m,2H),7.35(d,J＝7.8Hz,1H),2.53(s,3H).13CNMR(150MHz,Acetone-d6)δ198.22,165.64,159.92,150.09,144.90,140.10,139.72,138.38,138.17,135.96,132.14,131.48,130.71,129.35,129.20,127.59,126.77,126.55,125.96,121.76,111.35,27.62.HRMS(ESI):[M+Na]+calcd for C24H18ClN3NaO4S2 +534.0319,found 534.0323.HPLC purity,98％.
实施例42
化合物D10的合成步骤同化合物B16，以化合物11和4-(3-溴苯基)噻唑-2-胺制备得到，白色固体，产率61％。1H NMR(600MHz,Acetone-d6)δ8.31(d,J＝8.3Hz,2H),8.14(s,1H),8.02(d,J＝8.3Hz,2H),7.94(d,J＝7.8Hz,1H),7.84(s,1H),7.74(d,J＝7.6Hz,1H),7.71(s,1H),7.51(d,J＝7.9Hz,2H),7.45(t,J＝7.9Hz,1H),7.39(t,J＝7.9Hz,1H),2.53(s,3H).13CNMR(150MHz,Acetone-d6)δ198.19,165.64,159.94,149.97,144.92,140.12,139.73,138.64,138.18,132.43,132.33,131.48,130.72,130.58,129.22,126.77,126.56,126.36,124.17,121.77,111.36,27.62.HRMS(ESI):[M+H]+calcd for C24H19BrN3O4S2 +555.9995,found 555.9999.HPLCpurity,99％.
实施例43
化合物D11的合成步骤同化合物B16，以化合物11和4-(间甲苯基)噻唑-2-胺制备得到，白色固体，产率69％。1H NMR(600MHz,DMSO-d6)δ12.96(s,1H),10.75(s,1H),8.23(d,J＝8.5Hz,2H),7.92(d,J＝8.5Hz,2H),7.78(s,1H),7.73(d,J＝7.8Hz,1H),7.71–7.66(m,3H),7.44(t,J＝7.8Hz,1H),7.41–7.38(m,1H),7.33(t,J＝7.6Hz,1H),7.16(d,J＝7.5Hz,1H),2.51(s,3H),2.36(s,3H).13C NMR(150MHz,DMSO-d6)δ197.73,164.52,158.50,149.81,142.89,138.31,138.16,136.56,134.62,130.33,129.79,129.14,129.00,127.30,126.90,125.11,124.93,123.41,119.33,109.14,27.19,21.59.HRMS(ESI):[M-H]-calcd forC25H20N3O4S2 -490.0901,found 490.0917.HPLC purity,99％.
实施例44
化合物D12的合成步骤同化合物B16，以化合物11和4-(3-(三氟甲基)苯基)噻唑-2-胺制备得到，白色固体，产率53％。1H NMR(600MHz,DMSO-d6)δ13.01(s,1H),10.75(s,1H),8.30(s,1H),8.27-8.21(m,3H),7.99(s,1H),7.93(d,J＝8.5Hz,2H),7.73–7.65(m,4H),7.44(t,J＝7.8Hz,1H),7.41–7.38(m,1H),2.51(s,3H).13C NMR(150MHz,DMSO-d6)δ197.73,164.64,158.97,147.93,142.97,138.30,138.16,136.42,135.60,130.46,130.33,130.22,130.01,129.96,129.82,127.32,125.62,125.12,124.94,124.80,123.81,122.65,119.33,111.16,27.19.HRMS(ESI):[M-H]-calcd for C25H17F3N3O4S2 -544.0618,found544.0632.HPLC purity,97％.
实施例45
化合物D13的合成步骤同化合物B16，以化合物11和4-(3-甲氧基苯基)噻唑-2-胺制备得到，白色固体，产率63％。1H NMR(600MHz,DMSO-d6)δ12.98(s,1H),10.76(s,1H),8.24(d,J＝8.5Hz,2H),7.94(d,J＝8.5Hz,2H),7.77(s,1H),7.69(t,J＝4.2Hz,2H),7.52(dd,J＝8.3,5.0Hz,2H),7.44(t,J＝7.8Hz,1H),7.40(dd,J＝5.8,4.1Hz,1H),7.36(t,J＝7.9Hz,1H),6.92(dd,J＝8.1,2.2Hz,1H),3.82(s,3H),2.52(s,3H).13C NMR(150MHz,DMSO-d6)δ196.66,163.50,159.03,157.41,148.49,141.82,137.24,137.09,135.48,135.01,129.26,128.74,126.23,124.04,123.86,118.26,117.55,113.12,110.38,108.64,54.53,26.12.HRMS(ESI):[M-H]-calcdfor C25H20N3O5S2 -506.0850,found 506.0865.HPLC purity,99％.
实施例46
化合物D14的合成步骤同化合物B16，以化合物11和3-(2-氨基噻唑-4-基)苯酚制备得到，白色固体，产率27％。1H NMR(600MHz,Acetone-d6)δ8.32(d,J＝8.1Hz,2H),8.04(d,J＝8.4Hz,2H),7.86(s,1H),7.82–7.73(m,3H),7.54–7.50(m,1H),7.48–7.42(m,2H),7.19(dd,J＝8.0,2.2Hz,1H),7.04(s,1H),2.54(s,3H).13C NMR(150MHz,Acetone-d6)δ198.22,169.93,165.23,153.03,151.47,145.85,140.12,139.66,138.74,135.43,132.46,131.50,131.25,129.32,126.78,126.62,125.14,122.17,121.78,120.89,104.48,27.63.HRMS(ESI):[M+H]+calcd forC24H20N3O5S2 +494.0839,found 494.0845.HPLC purity,95％.
实施例47
化合物D15的合成步骤同化合物B16，以化合物11和4-(3-硝基苯基)噻唑-2-胺制备得到，白色固体，产率22％。1H NMR(600MHz,Acetone-d6)δ8.78(s,1H),8.36(d,J＝7.3Hz,1H),8.19(d,J＝8.1Hz,1H),8.03(d,J＝8.4Hz,2H),7.88–7.72(m,2H),7.76–7.71(m,2H),7.52(dd,J＝8.1,1.2Hz,1H),7.45(t,J＝7.9Hz,1H),2.53(s,3H).13C NMR(150MHz,Acetone-d6)δ196.40,163.95,158.50,148.88,147.46,143.19,138.32,137.94,136.29,131.58,130.09,129.69,128.96,127.44,125.00,124.77,122.25,120.46,119.99,110.66,25.82.HRMS(ESI):[M+Na]+calcd for C24H18N4NaO6S2 +545.0560,found 545.0568.HPLCpurity,99％.
实施例48
化合物D16的合成步骤同化合物B16，以化合物11和4-(4-氟苯基)噻唑-2-胺制备得到，白色固体，产率53％。1H NMR(600MHz,DMSO-d6)δ13.00(s,1H),10.76(s,1H),8.25–8.21(m,2H),8.01–7.96(m,2H),7.96–7.93(m,2H),7.72(s,1H),7.70–7.67(m,2H),7.44(t,J＝7.8Hz,1H),7.42–7.39(m,1H),7.32–7.26(m,2H),2.52(s,3H).13C NMR(150MHz,DMSO-d6)δ196.66,163.49,162.03,160.41,157.65,147.62,141.83,137.24,137.09,135.47,129.26,128.72,127.21,127.16,126.24,124.04,123.85,118.25,115.11,114.97,108.04,26.12.HRMS(ESI):[M-H]-calcd for C24H17FN3O4S2 -494.0650,found 494.0667.HPLCpurity,97％.
实施例49
化合物D17的合成步骤同化合物B16，以化合物11和4-(4-氯苯基)噻唑-2-胺制备得到，白色固体，产率49％。1H NMR(600MHz,DMSO-d6)δ12.98(s,1H),10.74(s,1H),8.22(d,J＝8.4Hz,2H),7.96(d,J＝8.5Hz,2H),7.93(d,J＝8.4Hz,2H),7.79(s,1H),7.70–7.66(m,2H),7.51(d,J＝8.5Hz,2H),7.43(t,J＝7.8Hz,1H),7.39(d,J＝8.3Hz,1H),2.51(s,3H).13CNMR(150MHz,DMSO-d6)δ197.75,164.61,158.82,148.47,142.97,138.34,138.18,136.51,133.58,132.82,130.32,129.78,129.27,127.96,127.31,125.08,124.95,119.36,110.06,27.18.HRMS(ESI):[M+H]+calcd for C24H19ClN3O4S2 +512.0500,found 512.0507.HPLCpurity,99％.
实施例50
化合物D18的合成步骤同化合物B16，以化合物11和4-(4-溴苯基)噻唑-2-胺制备得到，白色固体，产率49％。1H NMR(600MHz,DMSO-d6)δ12.99(s,1H),10.74(s,1H),8.23(d,J＝8.5Hz,2H),7.93(d,J＝8.5Hz,2H),7.90(d,J＝8.5Hz,2H),7.80(s,1H),7.68(d,J＝8.4Hz,2H),7.65(d,J＝8.5Hz,2H),7.44(t,J＝7.8Hz,1H),7.39(d,J＝8.3Hz,1H),2.51(s,3H).13CNMR(150MHz,DMSO-d6)δ197.72,164.59,158.82,148.52,142.97,138.32,138.19,136.50,133.93,132.18,130.32,129.78,128.26,127.31,125.08,124.94,121.43,119.37,110.15,27.18.HRMS(ESI):[M+H]+calcd for C24H19BrN3O4S2 +555.9995,found556.0005.HPLC purity,96％.
实施例51
化合物D19的合成步骤同化合物B16，以化合物11和4-(对甲苯基)噻唑-2-胺制备得到，白色固体，产率52％。1H NMR(600MHz,DMSO-d6)δ12.94(s,1H),10.73(s,1H),8.22(d,J＝8.4Hz,2H),7.92(d,J＝8.4Hz,2H),7.83(d,J＝8.0Hz,2H),7.70–7.66(m,2H),7.64(s,1H),7.43(t,J＝7.8Hz,1H),7.39(d,J＝8.4Hz,1H),7.25(d,J＝8.0Hz,2H),2.51(s,3H),2.33(s,3H).13C NMR(150MHz,DMSO-d6)δ197.72,164.50,158.48,149.81,142.90,138.32,138.18,137.67,136.58,132.07,130.32,129.79,129.77,127.29,126.19,125.08,124.94,119.37,108.46,27.18,21.28.HRMS(ESI):[M+H]+calcd for C25H22N3O4S2 +492.1046,found492.1057.HPLC purity,99％.
实施例52
化合物D20的合成步骤同化合物B16，以化合物11和4-(4-(三氟甲基)苯基)噻唑-2-胺制备得到，白色固体，产率26％。1H NMR(600MHz,Acetone-d6)δ8.32–8.28(m,2H),8.16(d,J＝8.1Hz,2H),8.02(d,J＝8.5Hz,2H),7.84(t,J＝1.9Hz,1H),7.81(s,1H),7.78(d,J＝8.3Hz,2H),7.76–7.74(m,1H),7.53–7.49(m,1H),7.45(t,J＝7.9Hz,1H),2.53(s,3H).13CNMR(150MHz,Acetone-d6)δ198.21,165.77,155.01,150.13,144.90,140.09,138.18,131.50,130.75,129.22,128.16,127.40,126.76,126.57,121.73,112.46,27.63.HRMS(ESI):[M+H]+calcd forC25H19F3N3O4S2 +546.0764,found 546.0773.HPLC purity,99％.
实施例53
化合物D21的合成步骤同化合物B16，以化合物11和4-(4-异丙基苯基)噻唑-2-胺制备得到，白色固体，产率63％。1H NMR(600MHz,DMSO-d6)δ12.96(s,1H),10.73(s,1H),8.22(d,J＝8.4Hz,2H),7.92(d,J＝8.4Hz,2H),7.86(d,J＝8.1Hz,2H),7.70–7.63(t,J＝12.7Hz,3H),7.43(t,J＝7.8Hz,1H),7.39(d,J＝8.5Hz,1H),7.31(d,J＝8.1Hz,2H),2.96–2.87(m,1H),1.23(d,J＝6.9Hz,6H).13C NMR(150MHz,DMSO-d6)δ197.73,164.51,158.50,149.83,148.60,142.91,138.34,138.18,136.59,132.44,130.31,129.78,127.28,127.13,126.26,125.07,124.94,119.37,108.53,33.66,27.18,24.27.HRMS(ESI):[M+H]+calcdfor C27H26N3O4S2 +520.1359,found 520.1364.HPLC purity,99％.
实施例54
化合物D22的合成步骤同化合物B16，以化合物11和4-(2,4-二氟苯基)噻唑-2-胺制备得到，白色固体，产率28％。1H NMR(600MHz,DMSO-d6)δ13.00(s,1H),10.74(s,1H),8.23(d,J＝8.4Hz,2H),8.13–8.07(m,1H),7.93(d,J＝8.4Hz,2H),7.68(d,J＝8.5Hz,2H),7.60(d,J＝2.1Hz,1H),7.47–7.37(m,3H),7.23(td,J＝8.5,2.3Hz,1H),2.51(s,3H).13C NMR(150MHz,DMSO-d6)δ197.73,164.67,161.21,159.18,158.27,142.98,142.59,138.32,138.19,136.49,131.03,130.32,129.81,127.30,125.08,124.95,119.37,113.26,113.17,112.56,112.42,105.17,27.18.HRMS(ESI):[M+H]+calcd for C24H18F2N3O4S2 +514.0701,found 514.0712.HPLC purity,99％.
实施例55
化合物D23的合成步骤同化合物B16，以化合物11和4-(3,4-二氟苯基)噻唑-2-胺制备得到，白色固体，产率44％。1H NMR(600MHz,Acetone-d6)δ8.29(d,J＝8.4Hz,2H),8.01(d,J＝8.4Hz,2H),7.89–7.82(m,2H),7.81–7.76(m,1H),7.74(d,J＝7.7Hz,1H),7.65(s,1H),7.53–7.49(m,1H),7.45(t,J＝7.9Hz,1H),7.38(dd,J＝18.9,8.5Hz,1H),2.53(s,3H).13C NMR(150MHz,Acetone-d6)δ198.22,165.73,160.05,152.92,152.38,151.29,150.75,149.52,144.93,140.12,139.75,138.21,134.10,131.49,130.72,129.21,126.78,126.56,124.20,121.77,119.45,116.60,27.62.HRMS(ESI):[M+H]+calcd for C24H18F2N3O4S2 +514.0701,514.0711.HPLC purity,96％.
测试例1
PHGDH抑制活性测试
以CBR-5884和BI-03-002作为阳性对照。
测试方法：将待测化合物粉末高速离心，12000rpm，2min。加入DMSO配成10mM母液暂时储存于-20℃冰箱。配制PHGDH酶反应缓冲液(200mM Tris，pH 8.1，400mM KCl和10mMEDTA)。NAD+配成24mM母液，3-PG配成100mM母液,心肌黄酶配成100unit/mL母液储存于-20℃冰箱待用。刃天青(Resazurin)配制成10mM母液储存于4℃冰箱待用(使用前稀释200倍)。之后该检测方法中所有化合物的稀释皆用PHGDH酶反应缓冲液。
待测化合物可按需先在96孔板中按梯度稀释好所需浓度。根据需要的体积配制PHGDH酶反应液(16ng/μL)，底物混合液(1mM 3-PG，50μM NAD+)。向384孔白色微孔反应板中，加入5μL含PHGDH酶缓冲液，2.5μL化合物，2.5μL的底物混合液，37℃条件下避光孵育60min。实验设置不加酶和不加抑制剂补充并相应体积酶反应液的孔作为Blank和Control。之后每孔加入5μL含有Diaphroase(0.25μM)和Resazurin(0.001unit/μL)的混合液，室温避光孵育10min。使用多功能酶标仪MD
在544nm激发波长和590nm发射波长下测量荧光信号。
测试结果如下表1所示：
表1
由表1数据可知，本申请提供的化合物对PHGDH具有抑制作用，通过A2与B1-B25的对比可知，用仲胺基取代(例如吗啉(B1)、硫代吗啉(B2)、1,1-二氧代-4-硫代吗啉(B3))导致抑制活性降低，IC50值超过20μM。然而，在哌啶对位引入羧基(B4)或乙氧基(B5)导致抑制活性略有增加，IC50值分别为12.96μM和12.21μM。对于具有非平面环的仲胺取代基，环己胺(B6)、4-氨基四氢吡喃(B7)和1-甲基哌啶-4-胺(B8)导致PHGDH抑制活性降低，而1-乙酰哌啶-四胺(B9)导致活性增加(IC50＝6.81±0.46μM)。然而，增加取代基的长度或柔韧性生成化合物B10-B14，其只能维持抑制活性，IC50值在10.85μM到17.84μM之间。对于芳胺的取代基，苯胺(B15)和4,6-二甲基嘧啶-2-胺(B16)分别提高了抑制活性，IC50值分别为6.50±0.80μM和3.19±1.00μM。2-氨基吡唑取代基(B17)导致抑制活性降低。在苯基的间位引入不同的取代基，例如羟甲基(B18)、羟乙基(B19)、吗啉(B20)、N-甲基哌嗪(B21)、羧基(B22)、羧甲基(B23)、乙酰基(B24)和N-甲基乙酰胺(B25)，其中化合物B18、B19、B20、B21、B23、B24的抑制活性与化合物B15相似。化合物B22和B25的抑制活性略有增加，IC50值分别为3.59±0.79μM和4.96±0.63μM。
通过比较化合物B24与C1-C6可以看出，4-苯基噻唑对于PHGDH抑制活性具有很大的影响。通过比较化合物B24与D1-D23可以看出在化合物B24的苯基上邻位、间位或对位引入不同取代基，可以看出2-氟(D1)、2-甲氧基(D5)、3-氟(D8)、3-甲基(D11)、3-甲氧基(D13)和4-氟(D16)的引入维持PHGDH抑制活性，IC50值范围为2.2μM至6.7μM。然而，其他化合物导致抑制活性显着降低。当同时引入两个氟原子时，与化合物D1、D8或D16相比，活性没有进一步增加(D22，IC50＝6.7±1.6μM；D23，IC50＝17.2±3.9μM)。
测试例2
抗MDA-MB-468和MDA-MB-231细胞增殖活性测试
MDA-MB-468为PHGDH基因扩增细胞系，MDA-MB-231为PHGDH低表达细胞系。
以BI-03-002、BI-01-013、CBR5884作为阳性对照。
测试方法：将培养皿中的细胞弃去上清培液，加入1mL PBS润洗后加胰酶于培养箱中消化，在显微镜下观察到大部分细胞被消化下来时加入相同体积的细胞培养基终止并用移液枪轻轻吹打成单个细胞，转入离心管中离心，1000rpm，离心5min。弃去上清后加入1-2mL培养基重悬，取100μL计数。根据所需种的密度于96孔板，每孔加入100μL培养基，单独设一个孔只加100μL培养基不种细胞作为Blank孔。第二天在96孔板中用无血清的培养基配制所需浓度的化合物，取10μL加入细胞中。对照组根据加药组中DMSO最高浓度配制含DMSO的培液取10μL加入细胞，Blank孔补充10μL培液。最外圈的孔由于边缘效应较大不用于实验。加药处理的96孔板放回细胞培养箱中继续培养，三天后收板，每孔加入10μL CCK8染液，用酶标仪在波长450nm处读板。4h内可多次读板，对照组的读值落在0.8～1.2范围内较为准确。实验设置2复孔。所有值扣除Blank后计算细胞存活率/抑制率：
存活率(％)＝【OD(给药孔)/OD(对照孔)】×100％
抑制率(％)＝100-存活率(％)
其中，SI＝对MDA-MB-231的IC50值/对MDA-MB-468的IC50值。
测试结果如下表2所示：
表2
如表2所示，多个化合物(B19、B21、B24、D5、D8、D11、D13)对PHGDH依赖性癌细胞(MDA-MB-468)的生长具有显着的抑制活性，并对人正常细胞MCF10A具有较好的选择性。
测试例3
体外代谢稳定性测试
测试方法：参照文献Gao,D.D.；Dou,H.X.；Su,H.X.；Zhang,M.M.；Wang,T.；Liu,Q.F.；Cai,H.Y.；Ding,H.P.；Yang,Z.；Zhu,W.L.；Xu,Y.C.；Wang,H.Y.；Li,Y.X.From Hit toLead:Structure-based Discovery of Naphthalene-1-sulfonamide Derivatives asPotent and SelectiveInhibitors of Fatty Acid Binding Protein4.Eur.J.Med.Chem.2018,154,44–59.公开的方法进行测试。
测试结果如下表3所示：
表3
如表3所示，化合物D8在所有三个物种中都显示出比其它三个化合物更好的肝微粒体稳定性。
测试例4
D8对PHGDH的抑制动力学测试
测试方法：
抑制动力学测试方法：设置不同浓度梯度的NAD+或3-PG，往384孔白色微孔反应板中加入5μL，酶与另一个底物混合按以上浓度配在同一管中，往384白色微孔反应板中加入5μL,最后Diaphroase和Resazurin的混合液，立即上机检测，此反应设置每2min测一次荧光值。最后需根据反应时间与荧光值计算反应速度，作酶反应速率与底物浓度的双倒数图。
为了表征PHGDH抑制剂D8的抑制机制，使用不同浓度的D8、3-PG和NAD+进行了竞争测试。Lineweaver-Burk图1表明，D8以与NAD+竞争的作用模式抑制PHGDH，NAD+是PHGDH的辅助因子。Lineweaver-Burk图2表明，D8对底物3-PG为非竞争模式。
测试例5
分子动力学模拟
测试方法：采用薛定谔Maestro 11.4软件中的Glide模块首先进行分子对接。(1)蛋白及格点文件准备：PHGDH蛋白的晶体复合物结构从Protein Data Bank上下载，PDB编号为2G76。使用Protein Preparation Wizard模块对蛋白进行加氢、添加缺失的侧链、调整蛋白质子化状态，删除B链、D-苹果酸及水分子，随后进行能量优化(OPLS2005力场，RMSD为
)。以NAD+为中心用Receptor Grid Generation模块制作格点文件，盒子大小均设置为
(2)化合物准备：将化合物通过LigPrep模块进行处理，输出相应的3D结构。(3)分子对接：分子对接在Ligand docking模块进行，采用超高精度模式(extraprecision mode)进行，选取打分最优的构象。(4)分子动力学模拟：采用SWISS-MODEL对人全长PHGDH进行同源建模，然后将上述对接最优构象在全长PHGDH蛋白结构中进行100ns的分子动力学模拟。
如图3和图4所示，100ns MD模拟后的最终模型。D8在cis-PHGDH和trans-PHGDH模型中是一致的。在20ns后，D8酰胺键的NH与D175大部分时间都存在氢键相互作用，距离为
图3中的d1和图4中的d2均表示化合物D8和D175氢键的距离。同时化合物D8中的磺酰基与G157存在弱相互作用。
测试例6
D8抑制MDA-MB-468细胞中丝氨酸合成测试
由于PHGDH是SSP中的一种重要酶，因此该蛋白质的抑制或缺乏将限制葡萄糖从头合成丝氨酸。
测试方法：取对数生长期的MDA-MB-468细胞适量种于六孔板中，每孔2mL DMEM/F-12培养基。24h后更换新鲜培养基并加入化合物GDD-04-35或DMSO预处理1h，随后用无菌的PBS溶液洗涤3次，吸净残留的液体后加入1mL含有15mM C13葡萄糖标记的三无DMEM培养基(补充1mM丙酮酸钠、4mM谷氨酰胺)同时补充相应浓度的化合物，放回培养箱培养2h后收集细胞代谢物。提前配置及预冷裂解液(甲醇:乙腈:水＝4:4:2，加内标1:150)，于-80℃冰箱预冷至少30min。每孔加入500μL裂解液，轻轻摇晃匀，使裂解液铺满孔底，于-80℃放置30min后在冰上用细胞刮板(碧云天)将细胞刮下收集于1.5mL EP管并用预冷的250μL裂解液润洗，合并洗液。Vortex 3次，每次30s，中间将样品放置冰上，保证样品的质量。随后4oC，14000g，离心15min，取上清500μL，放置于冷冻挥干机中挥干样品。将挥干的样品取出，每个样品加入40μL甲氧氨吡啶溶液复溶，涡旋3次，在37℃恒温摇床反应1h后加入衍生化试剂MtBSTFA 30μL，涡旋2次后用离心机轻甩下壁上的液体，在55℃恒温煮样器上反应1.5h。最后，2000rcf离心3min，取55μL上清加入进样瓶中，用GC-MS上级检测标记的丝氨酸比例。
测试结果如图5所示，采用t-Test检验，***p≤0.001；D8在10μM时显着降低了由13C-葡萄糖合成的丝氨酸，结果优于CBR-5884；表明MDA-MB-468细胞的SSP可以被化合物D8抑制。
测试例7
化合物D8对癌细胞系的抗增殖活性测试
PHGDH抑制剂可抑制PHGDH基因扩增(MDA-MB-468和BT-20)或PHGDH过表达(PC9、H1975、HCT116和HCC827)的细胞系增殖。因此，选择这些细胞系进行细胞增殖评估以证明D8的抗肿瘤活性与PHGDH有关。同时，使用PHGDH低表达细胞系(H23、Panc-1、MDA-MB-231、ZR-75-1和MCF-7)进行比较。
测试方法：将癌细胞置于含有10％FBS的适当培养基中的96孔板中过夜。之后，用一系列稀释的化合物处理细胞72小时。使用CCK-8细胞活力测定法(Dojindo，Japan)评估抗增殖活性。使用酶标仪测定450nm处的吸光度。
图6结果表明，D8对PHGDH基因扩增或过表达的细胞系具有显着敏感性，IC50值范围为3.5μM至21.6μM。然而，D8对PHGDH非依赖性细胞系的抗增殖活性降低，IC50值范围为28.6μM至39.2μM。这些抗增殖数据初步证明D8是一种潜在的PHGDH选择性抑制剂。
测试例8
药代动力学测试
由于化合物D8表现出优异的酶和细胞活性，因此使用口服(p.o.)和静脉内(i.v.)给药在ICR小鼠上评估其药代动力学特性。
测试方法：将六周大的ICR雄性小鼠圈养在18至29℃和30-70％相对湿度的12小时光照/12小时黑暗循环中。将小鼠随机分为三组，每组6只，体重21-25克。将化合物D8溶解在含DMSO/0.5％HPMC(5/95，v/v/)的水中，以3mg/kg的剂量口服给药；收集七个时间点(0.25、0.5、1.0、2.0、4.0、8.0和24小时)的血样。此外将化合物D8溶解在DMSO/EtOH/PEG300/0.9％NaCl(5/5/40/50,v/v/v/v)中，以1mg/kg的剂量静脉内给药；收集七个时间点(0.05、0.25、0.75、2.0、4.0、8.0和24小时)的血样。使用AQUITY UPLC分析D8的血浆浓度。
化合物D8的药代动力学测试结果如下表4所示，其中，AUC表示平均曲线下面积，T1/2表示平均半衰期，Tmax表示达峰时间，CL_obs表示清除率，Cmax表示最大血药浓度，F表示生物利用度。
表4
测试结果如表4和图7所示，对于1mg/kg剂量的静脉内给药，D8的平均半衰期为4.94小时，平均曲线下面积(AUC)值为38358h*ng/mL。以3mg/kg剂量口服D8后，它表现出最佳的平均半衰期(T1/2＝4.74h)、高药物暴露量(AUC last＝94386h*ng/mL)和良好的生物利用度(F％＝82.0％)。
测试例9
体内抗肿瘤测试
测试方法：将PC-9细胞悬浮在无血清培养基中，并将0.2mL含有5×106细胞注射到BALB/c裸鼠的右侧。用数字卡尺监测每只小鼠的肿瘤生长，并用方程计算：肿瘤体积＝0.5×长×宽2。当每只小鼠肿瘤体积达到50-100mm3时，将荷瘤裸鼠随机分为三个不同的治疗组(空白对照组0mg/kg，12.5mg/kg和25mg/kg；每组n＝6)。实验组每天腹腔注射一次化合物D8，连续31天。每周单独测量两次肿瘤大小。所有实验均根据动物伦理指南进行，并经上海药物研究所动物伦理和使用委员会批准。
如图8所示，one way ANOVA检验，*p<0.05，**p<0.01，D8表现出体内抗肿瘤作用并显着延缓肿瘤生长，TGI在12.5mg/kg时为41.4％，在25mg/kg时为68.9％。
如图9所示，剂量为25mg/kg的D8可显著减轻小鼠的肿瘤重量。如图10所示，D8在12.5mg/kg和25mg/kg两种剂量下均不会导致小鼠体重下降，这表明化合物D8具有安全性。
申请人声明，本发明通过上述实施例来说明本发明的工艺方法，但本发明并不局限于上述工艺步骤，即不意味着本发明必须依赖上述工艺步骤才能实施。所属技术领域的技术人员应该明了，对本发明的任何改进，对本发明所选用原料的等效替换及辅助成分的添加、具体方式的选择等，均落在本发明的保护范围和公开范围之内。
Claims (10)
1.一种对酰胺基苯磺酰基类化合物，其特征在于，所述对酰胺基苯磺酰基类化合物具有如下式I所示结构：
其中，R1选自卤素、取代或未取代的C6-C20芳基、取代或未取代的C2-C20杂芳基、取代或未取代的C1-C10直链或支链烷基、取代或未取代的C1-C10烷氧基、取代或未取代的C1-C10烷基羧基、取代或未取代的C3-C20环烷基、取代或未取代的C2-C20杂环烷基、取代或未取代的C2-C20烷基芳基、取代或未取代的C2-C20烷氧基芳基中的任意一种；
R2选自取代或未取代的C6-C20芳基、取代或未取代的C2-C20杂芳基、取代或未取代的C1-C10直链或支链烷基、取代或未取代的C3-C20环烷基、取代或未取代的C2-C20杂环烷基中的任意一种；
n选自0或1。
2.根据权利要求1所述对酰胺基苯磺酰基类化合物，其特征在于，所述取代的取代基各自独立地选自卤素、羧基、羟基、酰胺基、未取代或Ra取代的C1-C10直链或支链烷基、未取代或Ra取代的C1-C10烷氧基、未取代或Ra取代的C1-C10烷基羟基、未取代或Ra取代的C1-C10烷基羰基、未取代或Ra取代的C1-C10烷基酰胺基、未取代或Ra取代的C1-C10烷氧基酰胺基、NRN1RN2、未取代或Ra取代的C1-C10烷基羧基、未取代或Ra取代的C3-C20环烷基、未取代或Ra取代的C2-C20杂环烷基、未取代或Ra取代的C6-C20芳基、未取代或Ra取代的C2-C20杂芳基中的任意一种；
RN1、RN2各自独立地选自氢、C1-C10直链或支链烷基中的任意一种；
Ra选自卤素、羟基、硝基、C1-C10直链或支链烷基、C1-C10烷氧基或C1-C10卤代烷基中的任意一种。
3.根据权利要求1或2所述对酰胺基苯磺酰基类化合物，其特征在于，所述对酰胺基苯磺酰基类化合物选自如下式II-式VI所示结构中的任意一种：
其中，R11选自卤素、取代或未取代的C4-C20含氮杂环烷基中的任意一种；
R12选自取代或未取代的C3-C20环烷基、取代或未取代的C2-C20杂环烷基、取代或未取代的C6-C20芳基、取代或未取代的C2-C20杂芳基、取代或未取代的C1-C10直链或支链烷基、取代或未取代的C1-C10烷氧基、取代或未取代的C1-C10烷基羧基中的任意一种；
R2选自取代或未取代的C6-C20芳基、取代或未取代的C2-C20杂芳基中的任意一种；
p选自0-5的整数；X选自CH2、O或NH中的任意一种；L1选自单键、取代或未取代的C6-C20芳基、取代或未取代的C2-C20杂芳基中的任意一种；
q选自0-5的整数；Y选自CH2、羰基、O、NH、
中的任意一种；L2选自单键、取代或未取代的C6-C20芳基、取代或未取代的C2-C20杂芳基中的任意一种；
表示基团的连接位点。
4.根据权利要求1-3任一项所述对酰胺基苯磺酰基类化合物，其特征在于，所述R2选自取代的C2-C20杂芳基中的任意一种；
所述取代的C2-C20杂芳基的取代基选自未取代或Rb取代的C1-C10直链或支链烷基、未取代或Rb取代的C6-C20芳基中的任意一种；
所述Rb选自卤素、羟基、硝基、C1-C10直链或支链烷基、C1-C10烷氧基或C1-C10卤代烷基中的任意一种。
5.根据权利要求1-4任一项所述对酰胺基苯磺酰基类化合物，其特征在于，所述R2选自取代的噻唑基、取代的噻吩基、取代的三氮唑中的任意一种；
所述取代的取代基各自独立地选自未取代或Rc取代的苯基、未取代的C1-C10直链或支链烷基；
所述Rc选自卤素、羟基、硝基、C1-C10直链或支链烷基、C1-C10烷氧基或C1-C10卤代烷基中的任意一种。
6.根据权利要求3所述对酰胺基苯磺酰基类化合物，其特征在于，所述R11选自卤素、未取代或Rd取代的C4-C12含氮杂环烷基中的任意一种；
所述Rd选自羧基、C1-C10直链或支链烷基、C1-C10烷氧基中的任意一种；
优选地，所述R12选自未取代或Re取代的C2-C12杂环烷基、未取代或Re取代的C3-C12环烷基、未取代或Re取代的C6-C12芳基、未取代或Re取代的C2-C12杂芳基、未取代或Re取代的C1-C10直链或支链烷基、未取代或Re取代的C1-C10烷氧基、未取代或Re取代的C1-C10烷基羧基中的任意一种；
所述Re选自羟基、羧基、C1-C10直链或支链烷基、C1-C10烷氧基、C1-C10烷基羟基、C2-C12杂环烷基、C1-C10烷基羰基、C1-C10烷基酰胺基、C1-C10烷基羧基、NRN3RN4中的任意一种；
所述RN3、RN4各自独立地选自氢、C1-C10直链或支链烷基中的任意一种。
7.根据权利要求3所述对酰胺基苯磺酰基类化合物，其特征在于，所述L1、L2各自独立地选自单键、取代或未取代的C6-C12芳基、取代或未取代的C5-C12杂芳基中的任意一种。
8.根据权利要求1-7任一项所述对酰胺基苯磺酰基类化合物，其特征在于，所述对酰胺基苯磺酰基类化合物具有如下A2、B1-B25、C1-C6、D1-D23、E1-E5、F1-F7所示结构中的任意一种：
9.一种根据权利要求1-8任一项所述的对酰胺基苯磺酰基类化合物在制备磷酸甘油酸脱氢酶抑制剂中的应用。
10.一种根据权利要求1-8任一项所述的对酰胺基苯磺酰基类化合物在制备抗肿瘤药物中的应用。
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