含布洛芬-香豆素骨架化合物及其应用发明专利

专利名称：含布洛芬-香豆素骨架化合物及其应用

专利类型：发明专利

专利申请号：CN202211173554.2

专利申请（专利权）人：湖北医药学院
权利人地址：湖北省十堰市茅箭区人民南路30号

专利发明（设计）人：胡扬根,汤晓雨,王天帅,马俊凯

专利摘要：本发明公开了一类含布洛芬‑香豆素骨架化合物，其结构式如下：#imgabs0#此类化合物基于布洛芬和不同取代基的香豆素为药效骨架，以双酰肼、酰腙、噁二唑为键桥键接制得，制备方法简单，收率均在80％以上，初步抗肿瘤活性试验表明，该类化合物对HepG2肝癌细胞和Hela宫颈癌细胞的抑制活性表现较突出，化合物9e、9i、10d、10i、10e、10k对HepG2肝癌细胞和Hela宫颈癌细胞的活性均优于常用抗肿瘤药物吉非替尼，其中10e对HepG2肝癌细胞和Hela宫颈癌细胞的活性最优，镇痛试验表明该类化合物的镇痛作用优于布洛芬或与布洛芬相当。

主权利要求：
1.结构式如下表所示的含布洛芬‑香豆素骨架化合物在制备抗肿瘤药物中的应用；所述肿瘤类型为肝癌；
2.结构式如下表所示的含布洛芬‑香豆素骨架化合物在制备抗肿瘤药物中的应用；所述肿瘤类型为宫颈癌； 说明书 : 含布洛芬‑香豆素骨架化合物及其应用技术领域[0001] 本发明属于药物化学技术领域，具体涉及一种含布洛芬‑香豆素药效骨架化合物及其制备方法和应用。背景技术[0002] 肿瘤和疼痛对人类的健康和生存构成威胁，也大大降低生活质量，是世界面临的最重要的社会问题之一，尤其是在癌症的中期和晚期发展阶段。近年来，虽然临床上基于“一药一病”，对待肿瘤患者是同时使用化疗和止痛两种药物，即运用“鸡尾酒疗法”，取得了一定的疗效，但易产生耐药性、不同药物之间相互作用的不确定性等不足之处。因此，对于癌症和疼痛，研发结构多样、高效低毒的新型抗肿瘤和镇痛双重功效的药物势在必行。[0003] 在肿瘤发生、发展中炎症反应发挥着重要的作用，环氧合酶(COX)是影响炎症反应的重要靶点之一。布洛芬是一种常见的COX抑制剂，在临床上可用于治疗多种疾病，因其优秀的活性以及较高的安全性而被广泛应用。由于COX‑2在肿瘤细胞中大量表达，而布洛芬具有抑制COX‑2的作用。近年来，研究者们对布洛芬的分子结构进行修饰，所得到的化合物表现出一定的抗肿瘤活性。BaharehShokri实验组将布洛芬与已知的靶向多肽序列NGR偶联，其中NGR肽可以选择性地与肿瘤血管中过度表达的氨肽酶N靶向结合而发挥作用。抗肿瘤实验结果显示布洛芬的NGR偶联形式对SKOV‑3肿瘤细胞有较好的抑制作用。 的研究将α‑硫辛酸(ALA)与布洛芬通过共价键连接合成了一系列衍生物，并评估新合成的布洛芬衍生物对人类胶质母细胞瘤细胞的细胞毒性。结果显示，该类衍生物均能能够有效抑制人类胶质母细胞瘤细胞。[0004] 香豆素类化合物是一类重要的具有吡喃酮类含氧杂环化合物，具有高生物利用度和低毒性天然活性的先导结构骨架。基于其优秀的药理活性，一些香豆素的衍生物已被批准用于临床治疗。[0005] 化合物的拼合是指将两个或者多个药效基团利用化学键进行连接，进而得到新型的化合物分子。得到的新分子通常会继承参与拼合药效基团的药理活性，并同时降低药效基团的毒副作用。两种药效基团的拼合方式主要有“重叠式”和“链接式”两种类型。其中较常见的“链接式”主要是指将两种药效基团通过合适的连接基团杂交在一起，使用不同的连接基团进行拼合可以构成不同的连接策略。[0006] 目前未见与本发明相关化合物的报道。发明内容[0007] 本发明提供一类具有较好的抗肿瘤活性的含布洛芬‑香豆素骨架化合物，本发明利用香豆素天然的苯并吡喃酮环结构，利用双酰肼、噁二唑、酰腙键具有较大的偶极矩和氢键结合能力，将香豆素和布洛芬进行键接，形成了能与生物体内的多种酶及受体结合的含布洛芬‑香豆素骨架化合物。[0008] 本发明含布洛芬‑香豆素骨架化合物结构式如下：[0009][0010] 其中：Y选自 (双酰肼)、 (酰腙)、 (噁二唑)；[0011] R1选自H、卤素；[0012] R2选自H、卤素、CH3、OH、NO2、含有1‑3个碳原子的脂肪直链烷烃基、含有4‑7个碳原子的直链烷氧基、含有5‑6个碳原子的环烷氧基、1‑3个碳原子直链烷氧基取代的1‑3个碳原子的直链烷氧基，含有2‑4个碳原子的脂肪直链氨基、含有4‑6个碳原子的脂肪支链氨基；[0013] R3选自含有1‑3个碳原子的脂肪直链烷烃基、含有4‑7个碳原子的直链烷氧基、含有5‑6个碳原子的环烷氧基、1‑3个碳原子直链烷氧基取代的1‑3个碳原子的直链烷氧基，含有2‑4个碳原子的脂肪直链氨基、含有4‑6个碳原子的脂肪支链氨基；[0014] R4选自H、卤素。[0015] 本发明含布洛芬‑香豆素骨架化合物选自如下化合物：[0016][0017][0018][0019][0020] 上述含有布洛芬‑香豆素骨架化合物通过以下合成路线制得：化合物Ⅰ‑9a～Ⅰ‑9o的合成路线按照a‑b‑c合成步骤进行，化合物Ⅰ‑10a～Ⅰ‑10n的合成路线按照a‑e‑f合成步骤进行，化合物Ⅰ‑11a～Ⅰ‑11i的合成路线按照a‑b‑c‑d合成步骤进行；[0021][0022] (1)化合物Ⅰ‑9a～Ⅰ‑9o在溶剂存在、碱性条件下制备得到；[0023] 其中碱选用二甲基胺、二乙基胺、二丙基胺、吗啉中的一种；溶剂是二氯甲烷、甲醇、乙醇、乙腈、四氢呋喃、二氧六环中的一种或两种；[0024] (2)化合物Ⅰ‑10a～Ⅰ‑10n在酸性条件下制备得到；[0025] 其中酸选用醋酸、柠檬酸、苹果酸、酒石酸、乙酸、丁二酸和草酸中的一种；[0026] (3)化合物Ⅰ‑11a～Ⅰ‑11i由化合物Ⅰ‑9a～Ⅰ‑9o在脱水剂催化作用下发生脱水环化反应引入噁二唑，然后缚酸剂处理得到；[0027] 其中脱水剂有三氯化铝、三氯氧磷、五氧化二磷中的一种；缚酸剂为碳酸钾、碳酸氢钾、碳酸钠、碳酸氢钠、氢氧化钾、氢氧化钠、三乙胺、吡啶中的一种。[0028] 本发明另一目的是将上述含布洛芬‑香豆素骨架化合物应用在制备抗肿瘤药物中。[0029] 本发明另一目的是将上述含布洛芬‑香豆素骨架化合物应用在制备镇痛药物中。[0030] 本发明优点及技术效果：[0031] 以环氧化酶(COX)抑制剂布洛芬作为母体，根据药物的拼合原理，通过双酰肼、噁二唑、酰腙键与不同取代基香豆素骨架进行连接，合成一种新型的含有香豆素‑布洛芬骨架的化合物，制备方法简单，收率较高，收率80％以上；并对其抗肿瘤活性进行初步研究，体外活性试验表明，本发明提供的含有布洛芬‑香豆素骨架化合物有明显的抗肿瘤活性；镇痛试验表明该类化合物的镇痛作用优于布洛芬或与布洛芬相当，因此本发明化合物作为新型抗肿瘤和镇痛药物具有潜在的应用价值，对于新型抗肿瘤药物和镇痛药物的开发具有重要意义。具体实施方式[0032] 下面通过实施例对本发明作进一步详细说明，但本发明保护范围不局限于所述内容，实施例中方法如无特殊说明均为常规方法，使用的试剂，如无特殊说明均是常规市售试剂或按常规方法配制的试剂；[0033] 实施例1：化合物9a的制备[0034] 称取2.06g(10mmol)布洛芬酰肼、2.09g(10mmol)香豆素甲酰氯置于反应器中，然后加入20mL干燥的CH2Cl2作为溶剂，在冰浴条件下，边搅拌边缓慢滴入无水(C2H5)3N，直至体系的pH变为弱碱性，TLC法检测反应至原料全部消失，反应液减压浓缩除去溶剂，向浓缩液中加入60mL饱和食盐水，然后再加入60mL乙酸乙酯萃取，收集乙酸乙酯相，无水硫酸钠干燥，再浓缩，用硅胶(200‑300目)柱层析，石油醚‑乙酸乙酯混合液(体积比1:2)洗脱，收集洗脱液，浓缩、干燥得到目标化合物9a，产率：92％，白色固体。[0035] 1HNMR(400MHz,DMSO‑d6)δ:11.07(s,1H,NH),10.73(s,1H,NH),8.90(s,1H,H‑4coumarin),8.04‑7.11(m,8H,ArH),3.84‑3.83(m,1H,CH),2.42(d,J＝8.0Hz,2H,CH2),131.85‑1.80(m,1H,CH),1.41(d,J＝4.0Hz,3H,CH3),0.87(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,DMSO‑d6)δ:170.69,159.87,158.14,153.89,147.79,139.49,138.58,134.37,130.31,128.81,127.08,125.23,118.29,118.02,116.23,44.23,42.19,29.81,22.18,+ +18.42；HRMS(ESI)m/z:Anal.CalcdforC23H25N2O4([M+H]):393.1814,found:393.1833(M ,100)；Anal.RP‑UPLCtR＝24.588min,purity97.48％,UV330nm.[0036] 实施例2：化合物9b的制备[0037] 化合物9b的制备方法同实施例1，不同在于香豆素为6‑甲基香豆素甲酰氯，得到目标化合物9b，产率：96％，白色固体。[0038] 1HNMR(400MHz,CDCl3)δ:11.18(d,J＝8.0Hz,1H,NH),9.04(d,J＝8.0Hz,1H,NH),8.73(s,1H,H‑4coumarin),7.49‑7.10(m,7H,ArH),3.77‑3.72(m,1H,CH),2.45‑2.41(m,4H,1×CH1×CH3),1.87‑1.81(m,1H,CH),1.58(d,J＝8.0Hz,3H,CH3),0.89(d,J＝8.0Hz,136H,2×CH3)；CNMR(100MHz,CDCl3)δ:170.64,160.70,157.77,152.72,148.69,140.99,137.44,135.83,135.39,129.66,129.44,127.39,118.07,116.66,116.57,45.05,44.60,+30.18,22.42,20.77,18.55；HRMS(ESI)m/z:Anal.CalcdforC24H27N2O4([M+H] ):+407.1971,found:407.1970(M ,100)；Anal.RP‑UPLCtR＝25.447min,purity97.19％,UV330nm.[0039] 实施例3：化合物9c的制备[0040] 化合物9c的制备方法同实施例1，不同在于香豆素为7‑羟基香豆素甲酰氯，得到目标化合物9c，产率：82％，黄色固体。[0041] 1HNMR(400MHz,CDCl3)δ:11.02(s,1H,NH),10.63(s,1H,NH),8.82(s,1H,H‑4coumarin),7.96‑6.82(m,7H,ArH),3.82(t,J＝8.0Hz,1H,CH),2.41(m,2H,1×CH2),1.84‑131.79(m,1H,CH),1.39(d,J＝8.0Hz,3H,CH3),0.86(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,DMSO‑d6)δ:170.58,164.50,162.28,160.65,158.67,156.45,148.46,139.45,138.60,132.09,128.78,127.07,114.69,110.84,101.90,44.23,42.16,29.16,22.16,+ +18.41；HRMS(ESI)m/z:Anal.CalcdforC23H25N2O5([M+H]):409.1763,found:409.1764(M ,100)；Anal.RP‑UPLCtR＝22.876min,purity96.05％,UV330nm.[0042] 实施例4：化合物9d的制备[0043] 化合物9d的制备方法同实施例1，不同在于香豆素为6‑氯香豆素甲酰氯，得到目标化合物9d，产率：92％，白色固体。[0044] 1HNMR(400MHz,CDCl3)δ:11.05(d,J＝4.0Hz,1H,NH),8.74(s,1H,NH),8.40(d,J＝8.0Hz,1H,H‑4coumarin),7.65‑7.13(m,7H,ArH),3.71(q,J＝8.0Hz,1H,CH),2.46(d,J＝8.0Hz,2H,CH2),1.89‑1.82(m,1H,CH),1.61‑1.58(m,3H,CH3),0.91(d,J＝8.0Hz,6H,2×13CH3)；CNMR(100MHz,CDCl3)δ:170.53,159.93,157.07,152.81,147.47,141.26,137.07,134.51,130.90,129.82,128.82,127.40,119.24,118.36,117.95,45.04,44.88,30.19,+22.42,18.44；HRMS(ESI)m/z:Anal.CalcdforC23H24ClN2O4([M+H]):427.1425,found:+427.1424(M ,100)；Anal.RP‑UPLCtR＝25.773min,purity96.37％,UV330nm.[0045] 实施例5：化合物9e的制备[0046] 化合物9e的制备方法同实施例1，不同在于香豆素为6‑氟香豆素甲酰氯，得到目标化合物9e，产率：90％，白色固体。[0047] 1HNMR(400MHz,DMSO‑d6)δ:11.07(s,1H,NH),10.73(s,1H,NH),8.85(s,1H,H‑4coumarin),7.92‑7.09(m,7H,ArH),3.81(q,J＝8.0Hz,1H,CH),2.41(d,J＝8.0Hz,2H,13CH2),1.84‑1.77(m,1H,CH),1.38(d,J＝8.0Hz,3H,CH3),0.84(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,DMSO‑d6)δ:170.68,159.57,157.89,150.32,146.69,139.49,138.55,128.81,127.07,119.19,118.39,118.30,115.26,115.01,99.49,44.22,42.18,29.61,22.17,+18.42；HRMS(ESI)m/z:Anal.CalcdforC23H24FN2O4([M+H]):411.1720,found:411.1722(M+,100)；Anal.RP‑UPLCtR＝24.828min,purity98.92％,UV330nm.[0048] 实施例6：化合物9f的制备[0049] 化合物9f的制备方法同实施例1，不同在于香豆素为7‑丁氧基香豆素甲酰氯，得到目标化合物9f，产率：91％，白色固体。[0050] 1HNMR(400MHz,DMSO‑d6)δ:11.04(d,J＝8.0Hz,1H,NH),10.65(d,J＝4.0Hz,1H,NH),8.87(s,1H,H‑4coumarin),7.94‑7.05(m,7H,ArH),4.14(t,J＝8.0Hz,1H,CH),3.82(q,J＝8.0Hz,2H,CH2),2.42(d,J＝8.0Hz,2H,CH2),1.83‑1.71(m,3H,1×CH1×CH2),1.50‑1.43(m,2H,CH2),1.37(d,J＝8.0Hz,3H,CH3),0.95(t,J＝8.0Hz,3H,CH3)0.87(d,J＝8.0Hz,136H,2×CH3)；CNMR(100MHz,DMSO‑d6)δ:171.09,164.03,160.95,158.97,156.77,148.75,139.97,132.16,129.31,127.57,114.60,114.04,112.41,101.22,68.90,44.72,42.66,30.87,30.11,22.66,19.11,18.92,14.11；HRMS(ESI)m/z:Anal.CalcdforC27H33N2O5([M++ +H] ):465.2389,found:465.2393(M ,100)；Anal.RP‑UPLCtR＝28.293min,purity99.50％,UV330nm.[0051] 实施例7：化合物9g的制备[0052] 化合物9g的制备方法同实施例1，不同在于香豆素为6‑硝基香豆素甲酰氯，得到目标化合物9g，产率：84％，黄色固体。[0053] 1HNMR(400MHz,DMSO‑d6)δ:11.02(s,1H,NH),10.63(s,1H,NH),9.01‑7.10(m,8H,ArH),3.80(q,J＝8.0Hz,1H,CH),2.41(d,J＝8.0Hz,2H,CH2),1.84‑1.79(m,1H,CH),1.3913(d,J＝4.0Hz,CH3),0.86(d,J＝4.0Hz,6H,2×CH3)；CNMR(100MHz,DMSO‑d6)δ:170.89,158.78,157.91,157.30,146.41,143.94,139.51,138.54,128.82,128.31,127.08,126.00,120.44,118.68,117.84,44.22,42.24,29.61,22.16,18.41；HRMS(ESI)m/z:Anal.Calcd+ +forC23H24N3O6([M+H] ):438.1665,found438.1665(M ,100)；Anal.RP‑UPLCtR＝24.938min,purity99.64％,UV330nm.[0054] 实施例8：化合物9h的制备[0055] 化合物9h的制备方法同实施例1，不同在于香豆素为6,8‑二氯香豆素甲酰氯，得到目标化合物9h，产率：86％，黄色固体。[0056] 1HNMR(400MHz,DMSO‑d6)δ:11.05(s,1H,NH),10.64(s,1H,NH),8.83(s,1H,H‑4coumarin),8.15‑7.10(m,6H,ArH),3.80(q,J＝8.0Hz,1H,CH),2.42(d,J＝8.0Hz,2H,13CH2),1.85‑1.80(m,1H,CH),1.40(d,J＝8.0Hz,3H,CH3),0.87(d,J＝4.0Hz,6H,2×CH3)；CNMR(100MHz,DMSO‑d6)δ:170.82,158.47,157.73,148.32,146.10,139.50,138.52,132.95,128.81,128.74,128.20,127.08120.88,120.74,120.31,44.22,42.21,29.61,22.16,+18.41；HRMS(ESI)m/z:Anal.CalcdforC23H23Cl2N2O4([M+H]):461.1035,found:461.1034+(M ,100)；Anal.RP‑UPLCtR＝26.959min,purity99.15％,UV330nm.[0057] 实施例9：化合物9i的制备[0058] 化合物9i的制备方法同实施例1，不同在于香豆素为6,8‑二溴香豆素甲酰氯，得到目标化合物9i，产率：90％，白色固体。[0059] 1HNMR(400MHz,CDCl3)δ:10.09(s,1H,NH),8.69(s,1H,H‑4coumarin),8.27(s,1H,NH),8.02‑7.14(m,6H,ArH),3.70(q,J＝8.0Hz,1H,CH),2.46(d,J＝8.0Hz,2H,CH2),131.89‑1.82(m,1H,CH),1.60(d,J＝4.0Hz,3H,CH3),0.91(d,J＝4.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:170.63,158.98,156.70,150.28,147.13,141.32,139.84,136.99,131.09,129.86,127.42,120.45,118.62,118.06,111.49,45.04,44.96,30.19,22.42,+18.40；HRMS(ESI)m/z:Anal.CalcdforC23H23Br2N2O4([M+H]):549.0025,found:549.0024+(M ,100),UV330nm.[0060] 实施例10：化合物9j的制备[0061] 化合物9j的制备方法同实施例1，不同在于香豆素为7‑戊氧基香豆素甲酰氯，得到目标化合物9j，产率：94％，白色固体。[0062] 1HNMR(400MHz,CDCl3)δ:11.01(d,J＝4.0Hz,1H,NH),8.74(s,1H,H‑4coumarin),8.23(d,J＝8.0Hz,1H,NH),7.56‑6.84(m,7H,ArH),4.05(t,J＝8.0Hz,2H,CH2),3.70(q,J＝8.0Hz,1H,CH),2.46(d,J＝8.0Hz,2H,CH2),1.89‑1.80(m,3H,1×CH1×CH2),1.59(d,J＝138.0Hz,3H,CH3),1.47‑1.37(m,4H,2×CH2),0.96‑0.90(m,9H,3×CH3)；CNMR(100MHz,CDCl3)δ:170.45,164.97,158.38,156.92,148.69,141.18,137.20,130.99,129.80,127.42,114.70,112.76,111.97,110.86,69.10,45.05,44.95,30.18,28.56,28.05,22.42,+22.40,18.46,14.00；HRMS(ESI)m/z:Anal.CalcdforC28H35N2O5([M+H]):479.2546,+found:479.2543(M ,100)；Anal.RP‑UPLCtR＝29.468min,purity96.68％,UV330nm.[0063] 实施例11：化合物9k的制备[0064] 化合物9k的制备方法同实施例1，不同在于香豆素为7‑己氧基香豆素甲酰氯，得到目标化合物9k，产率：88％，白色固体。[0065] 1HNMR(400MHz,CDCl3)δ:11.02(d,J＝8.0Hz,1H,NH),8.74(s,1H,H‑4coumarin),8.38(d,J＝8.0Hz,1H,NH),7.55‑6.84(m,7H,ArH),4.05(t,J＝8.0Hz,2H,CH2),3.70(q,J＝8.0Hz,1H,CH),2.46(d,J＝8.0Hz,2H,CH2),1.89‑1.79(m,3H,1×CH1×CH2),1.63‑1.58(m,4H,2×CH2),1.49‑1.44(m,4H,2×CH2),1.36‑1.34(m,3H,CH3),0.93‑0.90(m,9H,3×CH3)；13CNMR(100MHz,CDCl3)δ:170.47,164.96,161.01,158.37,156.91,148.68,141.14,137.25,130.99,129.77,127.41,114.70,112.76,111.96,100.86,69.12,45.05,44.88,31.49,30.18,28.83,25.60,22.57,22.43,18.48,14.03；HRMS(ESI)m/z:Anal.Calcdfor+ +C29H37N2O5([M+H]):493.2702,found:493.2707(M ,100)；Anal.RP‑UPLCtR＝30.513min,purity99.92％,UV330nm.[0066] 实施例12：化合物9l的制备[0067] 化合物9l的制备方法同实施例1，不同在于香豆素为7‑环己氧基香豆素甲酰氯，得到目标化合物9，产率：86％，白色固体。[0068] 1HNMR(400MHz,CDCl3)δ:11.04(s,1H,NH),8.72(s,1H,H‑4coumarin),8.46(s,1H,NH),7.54‑6.84(m,7H,ArH),4.39‑4.34(m,1H,CH),3.71(q,J＝8.0Hz,1H,CH),2.45(d,J＝8.0Hz,2H,CH2),2.02‑1.66and1.46‑1.32(m,10H,5×CH21×CH3),1.59(d,J＝8.0Hz,133H,CH3),0.91(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:170.47,163.86,161.05,158.41,156.95,148.64,141.11,137.29,131.06,129.75,127.40,115.40,112.54,111.76,101.68,76.64,45.05,44.85,31.37,30.19,25.35,23.54,22.43,18.49；HRMS(ESI)m/z:+ +Anal.CalcdforC29H35N2O5([M+H]):491.2546,found:491.2549(M ,100)；Anal.RP‑UPLCtR＝29.335min,purity99.65％,UV330nm.[0069] 实施例13：化合物9m的制备[0070] 化合物9m的制备方法同实施例1，不同在于香豆素为7‑甲氧基乙氧基香豆素甲酰氯，得到目标化合物9m，产率：93％，白色固体。[0071] 1HNMR(400MHz,CDCl3)δ:11.01(d,J＝8.0Hz,1H,NH),8.83(s,1H,H‑4coumarin),8.49(d,J＝4.0Hz,1H,NH),7.56‑6.89(m,7H,ArH),4.21(t,J＝8.0Hz,1H,CH),3.71(q,J＝8.0Hz,H,CH),3.46(s,3H,CH3),2.45(d,J＝8.0Hz,2H,CH2),1.89‑1.83(m,1H,CH),1.59(d,J13＝4.0Hz,3H,CH3),0.91(d,J＝4.0Hz,3H,CH3)；CNMR(100MHz,CDCl3)δ:170.53,164.49,160.91,158.28,156.74,148.60,141.11,137.30,131.03,129.74,127.41,114.69,113.14,112.29,101.10,70.51,68.27,59.32,45.05,44.84,30.17,22.42,18.49；HRMS(ESI)m/z:+ +Anal.CalcdforC26H31N2O6([M+H]):467.2182；found:467.2182(M ,100)；Anal.RP‑UPLCtR＝24.565min,purity99.35％,UV330nm.[0072] 实施例14：化合物9n的制备[0073] 化合物9n的制备方法同实施例1，不同在于香豆素为6‑乙基香豆素甲酰氯，得到目标化合物9n，产率：94％，白色固体。[0074] 1HNMR(400MHz,CDCl3)δ:11.18(d,J＝8.0Hz,1H,NH),9.04(d,J＝8.0Hz,1H,NH),8.73(s,1H,H‑4coumarin),7.49‑7.10(m,7H,ArH),3.77‑3.72(m,1H,CH),2.7‑2.9(d,2H,CH2),2.45‑2.41(m,4H,1×CH1×CH3),1.87‑1.81(m,1H,CH),1.58(d,J＝8.0Hz,3H,CH3),130.89(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:170.64,160.70,157.77,152.72,148.69,140.99,137.44,135.83,135.39,129.66,129.44,127.39,118.07,116.66,116.57,45.05,44.60,30.18,22.42,20.77,18.55；HRMS(ESI)m/z:Anal.CalcdforC24H27N2O4([M++ +H] ):421.1971,found:420.1970(M ,100)；Anal.RP‑UPLCtR＝25.447min,purity97.19％,UV330nm.[0075] 实施例15：化合物9o的制备[0076] 化合物9o的制备方法同实施例1，不同在于香豆素为7‑环戊氧基香豆素甲酰氯，得到目标化合物9o，产率：89％，白色固体。[0077] 1HNMR(400MHz,CDCl3)δ:11.04(s,1H,NH),8.72(s,1H,H‑4coumarin),8.46(s,1H,NH),7.54‑6.84(m,7H,ArH),4.39‑4.34(m,1H,CH),3.71(q,J＝8.0Hz,1H,CH),2.45(d,J＝8.0Hz,2H,CH2),2.02‑1.66and1.46‑1.32(m,11H,4×CH21×CH3),1.59(d,J＝8.0Hz,133H,CH3),0.91(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:170.47,163.86,161.05,158.41,156.95,148.64,141.11,137.29,131.06,129.75,127.40,115.40,112.54,111.76,101.68,76.64,45.05,44.85,31.37,30.19,25.35,23.54,22.43,18.49；HRMS(ESI)m/z:+ +Anal.CalcdforC29H35N2O5([M+H]):479.1639,found:479.1639(M ,100)；Anal.RP‑UPLCtR＝29.335min,purity99.65％,UV330nm.[0078] 实施例16：化合物10a的制备[0079] 称取2.06g(10mmol)布洛芬酰肼，2.06g(10mmol)3‑乙酰基‑6‑氟香豆素置于反应器中，加入20mLEtOH作为溶剂，滴加数滴冰醋酸(HAc)作为催化剂，在60℃加热条件下回流搅拌反应，TLC法检测反应至原料全部消失，反应液减压浓缩除去溶剂，向浓缩液中加入60mL饱和碳酸氢钠水溶液，然后再加入60mL乙酸乙酯萃取，收集乙酸乙酯相，无水硫酸钠干燥，浓缩，用硅胶(200‑300目)柱层析洗脱，洗脱剂为石油醚和乙酸乙酯的混合液(石油醚:乙酸乙酯体积比＝1:3)，得到目标化合物10a，产率：89％，黄色固体。[0080] 1HNMR(400MHz,CDCl3)δ:8.67and8.34(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio33/67),8.08and7.57(s,1H,H‑4coumarin),7.33‑7.12(m,7H,ArH),4.56‑4.51and3.78‑3.76(m,1H,CH),2.48(d,J＝8.0Hz,2H,CH2),2.17and1.90(s,3H,CH3),1.89‑1.84(m,1H,CH),1.66and1.51(d,J＝138.0Hz,3H,CH3),0.91(d,J＝4.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:176.32,160.09,157.65,150.18,140.29,138.94,130.12,129.33,127.85,127.52,199.83,118.23,118.15,113.66,113.42,45.10,42.09,30.24,22.44,18.83,14.21；HRMS(ESI)m/z:Anal.Calcdfor+ +C24H26FN2O3([M+H]):409.1927,found:409.1931(M ,100)；Anal.RP‑UPLCtR(keto)＝25.147min,tR(enol)＝26.788min,purity99.62％,UV330nm.[0081] 实施例17：化合物10b的制备[0082] 制备工艺同实施例16，不同在于香豆素为3‑乙酰基‑7‑丁氧基香豆素，得到目标化合物10b，产率：95％，黄色固体。[0083] 1HNMR(400MHz,CDCl3)δ:8.71and8.36(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio35/65),8.09and7.64(s,1H,H‑4coumarin),7.41‑7.11(m,7H,ArH),4.59‑4.57and3.77‑3.76(m,1H,CH),4.06‑3.99(m,2H,CH2),2.49‑2.46(m,2H,CH2),2.17and1.91(s,3H,CH3),1.90‑1.80(m,3H,1×CH1×CH2),1.66and1.51(d,J＝4.0Hz,3H,CH3),1.57‑1.53and1.50‑1.47(m,2H,CH2),1.02‑130.97(m,3H,CH3),0.90(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:176.36,162.98,160.37,156.03,145.22,141.66,140.13,139.02,130.44,129.43,129.25,127.58,113.60,112.54,100.91,68.51,45.12,41.88,30.98,30.24,22.48,22.31,19.18,18.76,13.80；+ +HRMS(ESI)m/z:Anal.CalcdforC28H35N2O4([M+H]):463.2597,found:463.2598(M ,100)；Anal.RP‑UPLCtR(keto)＝28.818min,tR(enol)＝30.558min,purity99.97％,UV330nm.[0084] 实施例18：化合物10c的制备[0085] 制备工艺同实施例16，不同在于香豆素为3‑乙酰基‑7‑戊氧基香豆素，得到目标化合物10c，产率：96％，白色固体。[0086] 1HNMR(400MHz,CDCl3)δ:8.57and8.30(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio33/67),8.10and7.64(s,1H,H‑4coumarin),7.42‑6.77(m,7H,ArH),4.58‑4.56and3.77‑3.75(m,1H,CH),4.05‑3.99(m,2H,CH2),2.50‑2.46(m,2H,CH2),2.16and1.90(s,3H,CH3),1.88‑1.82(m,3H,1×CH1×CH2),1.66and1.51(d,J＝8.0Hz,3H,CH3),1.47‑1.38(m,4H,2×CH2),0.97‑0.94(m,3H,13CH3),0.90(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:176.26,163.00,160.37,156.04,141.69,140.15,139.00,130.07,129.44,129.27,127.58,122.90,113.62,112.54,100.91,68.82,45.12,41.90,30.24,28.66,28.10,22.42,22.31,18.75,14.26,14.00；HRMS+ +(ESI)m/z:Anal.CalcdforC29H37N2O4([M+H]):477.2753,found:477.2752(M ,100)；Anal.RP‑UPLCtR(keto)＝29.992min,tR(enol)＝31.758min,purity99.30％,UV330nm.[0087] 实施例19：化合物10d的制备[0088] 制备工艺同实施例16，不同在于香豆素为3‑乙酰基‑7‑(2‑甲氧基)乙氧基香豆素，得到目标化合物10d，产率：94％，白色固体。[0089] 1HNMR(400MHz,CDCl3)δ:8.63and8.35(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio34/66),8.09and7.64(s,1H,H‑4coumarin),7.42‑6.81(m,7H,ArH),4.58‑4.56and3.79‑3.78(m,1H,CH),4.21‑4.17(m,2H,CH2),3.81‑3.79(m,2H,CH2)2.50‑2.46(m,2H,CH2),2.17and1.91(s,3H,CH3),1.88‑131.85(m,1H,CH),1.66and1.51(d,J＝4.0Hz,3H,CH3),0.90(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:176.33,162.54,155.90,145.09,141.56,140.15,139.00,130.06,129.48,129.27,127.58,123.27,113.63,112.89,101.12,70.64,68.02,59.32,45.12,+41.89,30.23,22.45,18.75,14.29；HRMS(ESI)m/z:Anal.CalcdforC27H33N2O5([M+H]):+465.2389,found:465.2389(M ,100)；Anal.RP‑UPLCtR(keto)＝24.668min,tR(enol)＝26.070min,purity98.75％,UV330nm.[0090] 实施例20：化合物10e的制备[0091] 制备工艺同实施例16，不同在于香豆素为3‑乙酰基‑6,8‑二溴香豆素，得到目标化合物10e，产率：89％，白色固体。[0092] 1HNMR(400MHz,CDCl3)δ:8.92and8.43(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio30/70),8.00and7.89(s,1H,H‑4coumarin),7.59‑7.11(m,7H,ArH),4.53‑4.48and3.78‑3.76(m,1H,CH),2.49(d,J＝8.0Hz,2H,CH2),2.18and1.90(s,3H,CH3),1.89‑1.85(m,1H,CH),1.66and1.50(d,J＝134.0Hz,3H,CH3),0.91‑0.90(m,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:176.45,158.29,149.79,143.68,140.22,139.49,139.02,137.51,130.12,129.90,129.36,127.47,121.36,117.14,111.09,45.11,42.23,30.24,22.44,18.89,14.21；HRMS(ESI)m/z:Anal.Calcdfor+ +C24H25Br2N2O3([M+H] ):547.0232,found:547.0235(M ,100)；Anal.RP‑UPLCtR＝30.123min,purity99.54％,UV330nm.[0093] 实施例21：化合物10f的制备[0094] 制备工艺同实施例16，不同在于香豆素为3‑乙酰基‑7‑羟基香豆素，得到目标化合物10f，产率：87％，黄色固体。[0095] 1HNMR(400MHz,CDCl3)δ:9.77(s,1H,Ar‑OH),8.70and8.53(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio20/80),7.63and7.31(s,1H,H‑4coumarin),7.29‑6.96(m,7H,ArH),3.85‑3.80and3.76‑3.71(m,1H,CH),2.49(d,J＝8.0Hz,2H,CH2),2.16and1.92(s,3H,CH3),1.87‑1.84(m,H,CH),1.68and1.50(d,13J＝8.0Hz,3H,CH3),0.89(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:172.54,161.90,160.42,155.36,152.20,142.57,141.79,136.76,130.13,129.73,127.58,120.58,114.34,110.93,103.46,45.68,44.98,30.23,22.31,18.04,14.69；HRMS(ESI)m/z:+ +Anal.CalcdforC24H27N2O4([M+H]):407.1971,found:407.1970(M ,100)；Anal.RP‑UPLCtR(keto)＝22.742min,tR(enol)＝23.904min,purity99.40％,UV330nm.[0096] 实施例22：化合物10g的制备[0097] 制备工艺同实施例16，不同在于香豆素为3‑乙酰基‑7‑二乙基氨基香豆素，得到目标化合物10g，产率：97％，黄色固体。[0098] 1HNMR(400MHz,CDCl3)δ:8.77and8.41(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio37/63),8.04and7.62(s,1H,H‑4coumarin),7.31‑6.43(m,7H,ArH),4.63‑4.58and3.78‑3.76(m,1H,CH),3.47‑3.38(m,4H,2×CH2),2.46(d,J＝8.0Hz,2H,CH2),2.18and1.94(s,3H,CH3),1.88‑1.84(m,1H,CH),1.65and1.51(d,J＝4.0Hz,3H,CH3),1.27‑1.19(m,6H,2×CH3),0.90(d,J＝8.0Hz,6H,2×13CH3)；CNMR(100MHz,CDCl3)δ:176.36,161.15,156.96,151.22,142.09,140.01,139.14,129.94,129.67,129.20,127.63,118.98,109.29,108.51,97.00,45.13,44.93,41.77,+30.21,22.46,22.31,14.40,12.48；HRMS(ESI)m/z:Anal.CalcdforC28H36N3O3([M+H]):+462.2757,found:462.2760(M ,100)；Anal.RP‑UPLCtR(keto)＝27.011min,tR(enol)＝28.871min,purity99.24％,UV400nm.[0099] 实施例23：化合物10h的制备[0100] 制备工艺同实施例16，不同在于香豆素为3‑乙酰基‑6‑甲基香豆素，得到目标化合物10h，产率：94％，白色固体。[0101] 1HNMR(400MHz,CDCl3)δ:8.81and8.37(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio32/68),8.07and7.61(s,1H,H‑4coumarin),7.38‑7.12(m,7H,ArH),4.60‑4.54and3.78‑3.76(m,1H,CH),2.49‑2.47(m,2H,CH2),2.45‑2.39(m,3H,CH3),2.19and1.91(s,3H,CH3),1.88‑1.83(m,1H,CH),1.6613and1.51(d,J＝8.0Hz,3H,CH3),0.91(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:178.44,160.18,152.19,145.00,141.44,140.13,138.98,134.43,133.35,130.08,129.29,127.59,.126.66,118.82,116.31,45.14,41.92,30.25,22.48,20.83,18.79,14.40；HRMS+ +(ESI)m/z:Anal.CalcdforC25H29N2O3([M+H]):405.2178,found:405.2181(M ,100)；Anal.RP‑UPLCtR(keto)＝25.734min,tR(enol)＝27.580min,purity98.55％,UV330nm.[0102] 实施例24：化合物10i的制备[0103] 制备工艺同实施例16，不同在于香豆素为3‑乙酰基‑6,8‑二氯香豆素，得到目标化合物10i，产率：89％，白色固体。[0104] 1HNMR(400MHz,CDCl3)δ:8.66and8.35(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio34/66),8.04and7.61(s,1H,H‑4coumarin),7.47‑7.12(m,7H,ArH),4.53‑4.48and3.78‑3.76(m,1H,CH),2.49(d,J＝4.0Hz,2H,CH2),2.17and1.90(s,3H,CH3),1.90‑1.87(m,1H,CH),1.66and1.51(d,J＝138.0Hz,3H,CH3),0.91‑0.90(d,J＝4.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:176.24,148.27,139.60,132.22,132.06,130.16,129.91,129.37,128.50,127.47,126.50,126.15,122.53,120.89,104.48,45.09,42.22,30.25,22.45,18.89,14.05；HRMS(ESI)m/z:+ +Anal.CalcdforC24H25Cl2N2O3([M+H]):459.1242,found:459.1246(M ,100)；Anal.RP‑UPLCtR(keto)＝27.339min,tR(enol)＝29.455min,purity99.55％,UV330nm.[0105] 实施例25：化合物10j的制备[0106] 制备工艺同实施例16，不同在于香豆素为3‑乙酰基香豆素，得到目标化合物10j，产率：95％，白色固体。[0107] 1HNMR(400MHz,CDCl3)δ:8.85and8.33(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio34/66),8.14and7.68(s,1H,H‑4coumarin),7.59‑7.12(m,8H,ArH),4.57‑4.55and3.78‑3.76(m,1H,CH),2.50‑2.47(m,2H,CH2),2.18and1.91(s,3H,CH3),1.89‑1.83(m,1H,CH),1.66and1.51(d,J＝8.0Hz,133H,CH3),0.91(d,J＝4.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:176.31,159.95,154.03,144.68,141.41,140.22,138.94,132.29,130.10,129.30,128.47,127.58,124.76,119.08,116.63,45.12,41.98,30.26,22.46,22.32,18.79；HRMS(ESI)m/z:Anal.Calcdfor+ +C24H27N2O3([M+H]):391.2022,found:391.2025(M ,100)；Anal.RP‑UPLCtR(keto)＝24.710min,tR(enol)＝26.277min,purity99.55％,UV330nm.[0108] 实施例26：化合物10k的制备[0109] 制备工艺同实施例16，不同在于香豆素为3‑乙酰基‑6‑氯香豆素，得到目标化合物10k，产率：93％，白色固体。[0110] 1HNMR(400MHz,CDCl3)δ:8.65and8.42(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio29/71),8.33and8.05(s,1H,H‑4coumarin),7.64‑7.13(m,7H,ArH),4.55‑4.50and3.78‑3.76(m,1H,CH),2.49(d,J＝8.0Hz,2H,CH2),2.17and1.90(s,3H,CH3),1.87‑1.84(m,1H,CH),1.66and1.51(d,J＝138.0Hz,3H,CH3),0.91(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:176.28,159.33,152.34,144.08,140.04,138.96,134.28,132.13,130.00,129.35,127.81,127.62,127.50,120.10,118.20,118.02,30.25,22.45,18.85,17.99,14.18；HRMS(ESI)m/z:Anal.Calcd+ +forC24H26ClN2O3([M+H]):425.1632,found:425.1636(M ,100)；Anal.RP‑UPLCtR(keto)＝19.105min,tR(enol)＝28.065min,purity96.19％,UV330nm.[0111] 实施例27：化合物10l的制备[0112] 制备工艺同实施例16，不同在于香豆素为3‑乙酰基‑7‑丙基氨基香豆素，得到目标化合物10l，产率：94％，黄色固体。[0113] 1HNMR(400MHz,CDCl3)δ:8.77and8.41(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio37/63),8.04and7.62(s,1H,H‑4coumarin),7.31‑6.43(m,7H,ArH),4.63‑4.58and3.78‑3.76(m,1H,CH),3.47‑3.38(m,4H,2×CH2),2.46(d,J＝8.0Hz,2H,CH2),2.18and1.94(s,3H,CH3),1.88‑1.84(m,1H,CH),131.64(d,J＝4.0Hz,3H,CH3),1.27‑1.19(m,6H,2×CH3),0.90(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:176.36,161.15,156.96,151.22,142.09,140.01,139.14,129.94,129.67,129.20,127.63,118.98,109.29,108.51,97.00,45.13,44.93,41.77,30.21,+22.46,22.31,14.40,12.48；HRMS(ESI)m/z:Anal.CalcdforC28H36N3O3([M+H] ):+447.2757,found:447.2760(M ,100)；Anal.RP‑UPLCtR(keto)＝27.011min,tR(enol)＝28.871min,purity99.24％,UV400nm.[0114] 实施例28：化合物10m的制备[0115] 制备工艺同实施例16，不同在于香豆素为3‑乙酰基‑7‑二丙基氨基香豆素，得到目标化合物10m，产率：90％，黄色固体。[0116] 1HNMR(400MHz,CDCl3)δ:8.77and8.41(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio37/63),8.04and7.62(s,1H,H‑4coumarin),7.31‑6.43(m,7H,ArH),4.63‑4.58and3.78‑3.76(m,1H,CH),3.47‑3.38(m,4H,2×CH2),2.87‑3.08(m,4H,2×CH2),2.46(d,J＝8.0Hz,2H,CH2),2.18and1.94(s,3H,CH3),1.88‑1.84(m,1H,CH),1.65and1.51(d,J＝4.0Hz,3H,CH3),1.27‑1.19(m,6H,2×13CH3),0.90(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:176.36,161.15,156.96,151.22,142.09,140.01,139.14,129.94,129.67,129.20,127.63,118.98,109.29,108.51,97.00,45.13,44.93,41.77,30.21,22.46,22.31,14.40,12.48；HRMS(ESI)m/z:Anal.Calcd+ +forC28H36N3O3([M+H]):490.1548,found:490.1548(M ,100)；Anal.RP‑UPLCtR(keto)＝27.011min,tR(enol)＝28.871min,purity99.24％,UV400nm.[0117] 实施例29：化合物10n的制备[0118] 制备工艺同实施例16，不同在于香豆素为3‑乙酰基‑7‑乙基氨基香豆素，得到目标化合物10n，产率：92％，黄色固体。[0119] 1HNMR(400MHz,CDCl3)δ:8.77and8.41(s,1H,NHandOH,keto‑enoltautomerism,keto(‑CONH‑)/enol(‑HOC＝N‑)ratio37/63),8.04and7.62(s,1H,H‑4coumarin),7.31‑6.43(m,7H,ArH),4.63‑4.58and3.78‑3.76(m,1H,CH),3.47‑3.38(m,4H,2×CH2),2.46(d,J＝8.0Hz,2H,CH2),2.18and1.94(s,3H,CH3),1.88‑1.84(m,1H,CH),131.64(d,J＝4.0Hz,3H,CH3),1.27‑1.19(m,6H,2×CH3),0.90(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:176.36,161.15,156.96,151.22,142.09,140.01,139.14,129.94,129.67,129.20,127.63,118.98,109.29,108.51,97.00,45.13,44.93,41.77,30.21,+22.46,22.31,14.40,12.48；HRMS(ESI)m/z:Anal.CalcdforC28H36N3O3([M+H] ):+434.2399,found:433.2365(M ,100)；Anal.RP‑UPLCtR(keto)＝27.011min,tR(enol)＝28.871min,purity99.24％,UV400nm.[0120] 实施例30：化合物11a的制备[0121] 称取4.10g(10mmol)实施例5得到的化合物9e，加入5mL三氯氧磷，在80℃加热条件下回流搅拌反应6h，TLC法检测反应至原料全部消失，冷却至室温，依次加入饱和食盐水、碳酸氢钠饱和水溶液搅拌5min，然后加入50mL乙酸乙酯萃取，收集乙酸乙酯相，无水硫酸钠干燥，在浓缩，用柱层析洗脱，柱层析硅胶为200‑300目，洗脱剂为石油醚和乙酸乙酯的混合液(石油醚:乙酸乙酯＝1:3)，得到目标化合物11a，产率：90％，白色固体。[0122] 1HNMR(400MHz,CDCl3)δ:8.48(s,H,H‑4coumarin),7.39‑7.11(m,7H,ArH),4.46(q,J＝8.0Hz,H,CH),2.45(d,J＝4.0Hz,2H,CH2),1.87‑1.86(m,1H,CH),1.82(d,J＝8.0Hz,133H,CH3),0.89(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:170.28,160.70,155.89,150.75,143.68,141.18,137.14,.129.68,127.10,121.69,121.45,118.67,118.59,114.31,114.07,45.02,37.18,30.17,22.38,19.70；HRMS(ESI)m/z:Anal.Calcdfor+ +C23H22FN2O3([M+H]):393.1614,found:393.1613(M ,100)；Anal.RP‑UPLCtR＝26.713min,purity99.27％,UV330nm.[0123] 实施例31：化合物11b的制备[0124] 采用4.27g(10mmol)实施例4制得的化合物9d为原料，反应过程同实施例30，得到目标化合物11b，产率：93％，白色固体。[0125] 1HNMR(400MHz,CDCl3)δ:8.48(s,H,H‑4coumarin),7.60‑7.11(m,7H,ArH),4.46(q,J＝8.0Hz,H,CH),2.45(d,J＝4.0Hz,2H,CH2),1.87‑1.84(m,1H,CH),1.82(d,J＝8.0Hz,133H,CH3),0.89(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:170.28,160.63,155.86,152.89,143.36,141.19,137.11,133.89,130.56,129.68,128.18,127.10,118.96,118.40,113.98,45.02,37.18,30.17,22.39,19.89；HRMS(ESI)m/z:Anal.CalcdforC23H22ClN2O3+ +([M+H]):409.1319,found:409.1322(M ,100)；Anal.RP‑UPLCtR＝27.959min,purity99.20％,UV330nm.[0126] 实施例32：化合物11c的制备[0127] 采用4.61g(10mmol)实施例8得到的化合物9h为原料，反应过程同实施例30，得到目标化合物11c，产率：86％，白色固体。[0128] 1HNMR(400MHz,CDCl3)δ:8.45(s,H,H‑4coumarin),7.69‑7.12(m,6H,ArH),4.46(q,J＝8.0Hz,H,CH),2.45(d,J＝4.0Hz,2H,CH2),1.87‑1.86(m,1H,CH),1.82(d,J＝8.0Hz,133H,CH3),0.89(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:170.52,160.29,154.54,148.83,142.90,141.24,137.01,133.73,130.38,129.69,127.10,126.70,123.01,119.75,114.71,45.02,37.19,30.17,22.37,19.66；HRMS(ESI)m/z:Anal.CalcdforC23H21Cl2N2O3+ +([M+H]):443.0929,found:443.0933(M ,100)；Anal.RP‑UPLCtR＝29.108min,purity97.29％,UV330nm.[0129] 实施例33：化合物11d的制备[0130] 采用5.50g(10mmol)实施例9得到的化合物9i为原料，反应过程同实施例30，得到目标化合物11d，产率：90％，白色固体。[0131] 1HNMR(400MHz,CDCl3)δ:8.42(s,H,H‑4coumarin),7.98‑7.11(m,6H,ArH),4.46(q,J＝8.0Hz,H,CH),2.45(d,J＝4.0Hz,2H,CH2),1.87‑1.84(m,1H,CH),1.82(d,J＝8.0Hz,133H,CH3),0.89(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:170.49,160.25,154.83,150.36,142.81,141.24,139.27,137.02,130.44,129.69,127.11,120.19,117.68,114.61,111.54,45.02,37.19,30.17,22.38,19.55；HRMS(ESI)m/z:Anal.CalcdforC23H21Br2N2O3+ +([M+H]):530.9919,found:530.9921(M ,100)；Anal.RP‑UPLCtR＝29.544min,purity98.94％,UV330nm.[0132] 实施例34：化合物11e的制备[0133] 采用4.64g(10mmol)实施例6得到的化合物9f为原料，反应过程同实施例30，得到目标化合物11e，产率：91％，白色固体。[0134] 1HNMR(400MHz,CDCl3)δ:8.45(s,H,H‑4coumarin),7.50‑6.83(m,7H,ArH),4.45(q,J＝8.0Hz,H,CH),4.06(t,J＝8.0Hz,2H,CH2),2.44(d,J＝8.0Hz,2H,CH2),1.87‑1.84and1.80‑1.78(m,1H,CH),1.81(d,J＝8.0Hz,3H,CH3),1.56‑1.47(m,2H,CH2),0.99(t,J＝138.0Hz,3H,CH3),0.89(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:169.67,164.47,161.45,156.89,156.79,144.88,141.04,137.40,130.24,129.62,127.10,114.20,111.61,108.76,101.11,68.73,45.03,37.17,30.92,30.15,22.38,19.76,19.15,13.75；HRMS(ESI)+ +m/z:Anal.CalcdforC27H31N2O4([M+H]):447.2284,found:447.2288(M ,100)；Anal.RP‑UPLCtR＝30.316min,purity98.05％,UV330nm.[0135] 实施例35：化合物11f的制备[0136] 采用4.06g(10mmol)实施例2得到的化合物9b为原料，反应过程同实施例30，得到目标化合物11f，产率：92％，白色固体。[0137] 1HNMR(400MHz,CDCl3)δ:8.47(s,H,H‑4coumarin),7.45‑7.11(m,7H,ArH),4.46(q,J＝8.0Hz,H,CH),2.45‑2.43(m,5H,1×CH21×CH3),1.87‑1.84(m,1H,CH),1.82(d,J＝134.0Hz,3H,CH3),0.89(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:170.01,161.15,156.58,152.78,144.89,141.11,137.27,135.20,135.04,129.65,128.82,127.12,117.78,116.66,112.65,45.03,37.17,30.17,22.39,20.75,19.74；HRMS(ESI)m/z:Anal.Calcdfor+ +C24H25N2O3([M+H]):389.1865,found:389.1867(M ,100)；Anal.RP‑UPLCtR＝27.415min,purity99.83％,UV330nm.[0138] 实施例36：化合物11g的制备[0139] 采用3.92g(10mmol)实施例1得到的化合物9a为原料，反应过程同实施例30，得到目标化合物11g，产率：87％，白色固体。[0140] 1HNMR(400MHz,CDCl3)δ:8.53(s,H,H‑4coumarin),7.66‑7.11(m,8H,ArH),4.46(q,J＝8.0Hz,H,CH),2.45(d,J＝4.0Hz,2H,CH2),1.87‑1.84(m,1H,CH),1.82(d,J＝8.0Hz,133H,CH3),0.89(d,J＝8.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:170.06,161.01,156.29,154.60,144.81,141.12,137.26,134.02,129.66,129.19,127.10,125.16,118.03,116.95,112.88,45.03,37.18,30.15,22.38,19.73；HRMS(ESI)m/z:Anal.CalcdforC23H23N2O3([M++ +H] ):375.1709,found:375.1709(M ,100)；Anal.RP‑UPLCtR＝26.348min,purity99.49％,UV330nm.[0141] 实施例37：化合物11h的制备[0142] 采用4.78g(10mmol)实施例10得到的化合物9j为原料，反应过程同实施例30，得到目标化合物11，产率：86％，黄色固体。[0143] 1HNMR(400MHz,CDCl3)δ:8.44(s,H,H‑4coumarin),7.50‑6.82(m,7H,ArH),4.45(q,J＝8.0Hz,H,CH),4.06(t,J＝8.0Hz,2H,CH2),2.44(d,J＝8.0Hz,2H,CH2),1.92‑1.83(m,3H,1×CH1×CH2),1.81(d,J＝8.0Hz,3H,CH3),1.49‑1.37(m,4H,2×CH2),0.94(t,J＝138.0Hz,3H,CH3),0.89(d,J＝4.0Hz,6H,2×CH3)；CNMR(100MHz,CDCl3)δ:169.60,164.46,161.42,156.84,156.76,144.87,141.01,137.41,130.29,129.61,127.10,114.18,111.58,108.67,101.9,69.03,45.02,37.14,30.15,28.58,28.07,22.39,19.77,13.99；HRMS(ESI)+ +m/z:Anal.CalcdforC28H33N2O4([M+H]):461.2440,found:461.2443(M ,100)；Anal.RP‑UPLCtR＝31.467min,purity99.89％,UV330nm.[0144] 实施例38：化合物11i的制备[0145] 采用4.92g(10mmol)实施例11得到的化合物9k为原料，反应过程同实施例30，得到目标化合物11i，产率：94％，白色固体。[0146] 1HNMR(400MHz,CDCl3)δ:8.45(s,H,H‑4coumarin),7.50‑6.82(m,7H,ArH),4.44(q,J＝8.0Hz,H,CH),4.05(t,J＝8.0Hz,2H,CH2),2.44(d,J＝8.0Hz,2H,CH2),1.87‑1.75(m,6H,1×CH1×CH21×CH3),1.49‑1.44(m,2H,CH2),1.37‑1.35(m,4H,2×CH2),0.93‑0.8813(m,9H,3×CH3)；CNMR(100MHz,CDCl3)δ:169.64,164.46,161.44,156.87,156.76,144.86,141.02,137.41,130.25,129.61,127.10,114.19,111.60,108.72,101.11,69.05,45.03,37.16,31.49,30.14,28.86,25.59,22.55,22.38,19.77,13.99；HRMS(ESI)m/z:+ +Anal.CalcdforC29H35N2O4([M+H]):475.2597,found:475.2592(M ,100)；Anal.RP‑UPLCtR＝32.963min,purity99.35％,UV330nm.[0147] 实施例39：CCK‑8法检测上述实施例制得的含有布洛芬‑香豆素骨架化合物的抗肿瘤活性[0148] 取对数生长期的细胞，弃去旧的培养基，用PBS清洗三次以洗净培养基及细胞代谢废物，然后加入1mL0.25％胰蛋白酶，37℃下消化1‑2min，消化完毕，加入1‑2mL完全培养基终止消化，并用移液枪轻轻将贴壁细胞吹打下来，转移至15mL离心管内，1000r/min离心5分钟，离心完毕弃去上清，加入1‑2mL完全培养基，通过移液枪吹打使细胞悬浮，吸取10μL细胞悬液于细胞计数板上，盖上盖玻片，在显微镜下进行计数。在96孔板中心55个孔中接种上一步制备好的细胞悬液(100μL/孔)，同时设置5个孔只添加培养基作为空白组，在37℃、5％CO2培养箱中培养24h，之后吸去培养基，加入PBS清洗，加入含有5种不同浓度受试化合物的培养基，每种浓度设立5个复孔，设置5个接种过细胞的复孔只加培养基作为对照组；加完药后培养48h，弃去孔内培养基，向每个孔中加入含有10μL增强型CCK‑8溶液的培养基，转入培养箱中在37℃、5％CO2条件下孵育4h；使用酶标仪测量其在450nm处的吸光度(A)，重复测量三次取其平均值。[0149] 某种浓度下受试化合物的抑制率＝[1‑(A药物‑A空白)/(A对照‑A空白)]×100％；将浓度和抑制率数据导入SPSS软件中，即可算出该药物的IC50值；[0150] 通过CCK‑8法检测了化合物对HepG2细胞以及Hela细胞的增殖抑制作用，检测结果见表1、表2、表3：[0151] 表1化合物(9a‑9m)对HepG2和Hela的增殖抑制活性[0152][0153][0154] 表2化合物(10a‑10k)对HepG2和Hela的增殖抑制活性[0155][0156][0157] 表3化合物(11a‑11i)对HepG2和Hela的增殖抑制活性[0158][0159] 从表中可以看出：以吉非替尼为阳性对照，本发明含布洛芬‑香豆素骨架化合物对HepG2肝癌细胞和Hela宫颈癌细胞都有潜在的抗肿瘤活性，9e、9g、9i、10d、10e、10f、10h、10i、10k、11g活性较好，其中10e最为优异，抑制两种癌细胞的活性均优于布洛芬和常用抗肿瘤药物吉非替尼。[0160] 实施例40：醋酸扭体法检测上述实施例制得的含布洛芬‑香豆素骨架化合物的镇痛活性[0161] 取24只小鼠，随机分成阴性对照组、阳性对照组、给药组，每组8只，给药组按照小鼠体重2mg/10g给药剂量对小鼠灌胃化合物溶液(4mg/mL)，阳性对照组按照小鼠体重1mg/10g的给药剂量对小鼠灌胃布洛芬溶液(4mg/mL)，阴性对照组小鼠灌胃等量生理盐水，1.5h后腹腔注射0.7％冰醋酸0.1mL/10g，观察记录小鼠在注射冰醋酸后20min内的扭体次数，平行做三次，计算镇痛率；[0162] 镇痛率(％)＝(阴性对照组小鼠扭体反应次数‑给药组小鼠扭体反应次数)/阴性对照组小鼠扭体反应次数×100％，结果见表4；[0163] 表4化合物镇痛率[0164] 化合物 标准差 t检验 镇痛率9a 5.9462 0.01103015 36.25％9d 6.8661 0.00066495 80.00％9f 6.3189 0.03007355 31.25％9g 5.2576 0.0367161 26.25％9h 8.2980 0.76018587 5.00％9i 9.5160 0.01226017 61.88％9j 4.2426 0.00018658 60.00％9m 8.0133 0.08670649 28.75％10a 8.6879 0.00710610 57.71％10b 6.1818 0.39207902 11.25％10e 5.8172 0.00011615 84.38％10g 6.3310 0.00727993 57.30％10h 6.4904 0.02088597 34.38％11b 6.0223 0.35814179 ‑11.88％11e 5.6315 0.07987136 22.50％11f 4.9785 0.02013302 28.75％11h 8.3313 0.01145093 56.88％生理盐水 3.7033 ‑‑ ‑‑‑7布洛芬 3.2514 1.26574×10 85.00％[0165] 从表中数据可以看出：本发明化合物均具有镇痛活性，其中10e和9d的镇痛效果较好与布洛芬镇痛效果相当。

专利地区：湖北

专利申请日期：2022-09-26

专利公开日期：2024-11-19

专利公告号：CN115368333B