一种喹唑啉酮与α，β-不饱和酮轭合衍生物及其制备方法和应用转让专利
申请号 : CN201710658952.6
文献号 : CN107522700B
文献日 : 2020-03-20
发明人 : 黄志纾 , 谭嘉恒 , 王玉青 , 王晨曦
申请人 : 中山大学
摘要 :
本发明公开了一种喹唑啉酮与α，β‑不饱和酮轭合衍生物，所述喹唑啉酮与α，β‑不饱和酮轭合衍生物的结构式如式(I)或式(II)所示：其中，R1为氢、胺基、取代胺基、五元或六元杂环基；R2为氢、苯基、取代苯基、苯并杂环化合物；取代基选自C1‑5烷基、C1‑8烷氧基、卤基、苯基、胺基、五元或六元杂环基。本发明提供的喹唑啉酮与α,β‑不饱和酮轭合衍生物可以有效抑制多种肿瘤细胞的生长，并能够显著抑制原癌基因c‑MYC的转录和表达，在制备抗肿瘤药物上有着广阔的应用空间。
权利要求 :
1.一种喹唑啉酮与α，β-不饱和酮轭合衍生物，其特征在于，所述喹唑啉酮与α，β-不饱和酮轭合衍生物的结构式如式(I)所示：其中，R1为NH(CH2)nR3，
R3为N,N-二乙基甲胺基或吗啉基；n为1～5中任意一个整数；
R2为取代苯基或1,3-苯并二噁茂，取代苯基中取代基为氟、吗啉基、叔丁基或甲氧基。
2.一种权利要求1中所述的喹唑啉酮与α,β-不饱和酮轭合衍生物的制备方法，其特征在于，包括如下步骤：S1.2-氨基-4,5-二氟苯甲酸与2-吡咯烷酮反应得到化合物S2.化合物 与胺类化合物反应得到中间体
S3.化合物 与酮类化合物反应得到中间体
S4.将S2中所得化合物与S3中所得化合物反应，得到所述喹唑啉酮与α,β-不饱和酮轭合衍生物。
3.权利要求1所述的喹唑啉酮与α,β-不饱和酮轭合衍生物在制备抗癌药物中的应用。
4.根据权利要求3所述的应用，其特征在于，所述喹唑啉酮与α,β-不饱和酮轭合衍生物是作为原癌基因c-MYC转录调控阻断剂应用于抗癌药物的制备。
5.根据权利要求3所述的应用，其特征在于，所述抗癌为抗卵巢癌、宫颈癌、乳腺癌、肺腺癌、结肠癌、肝癌、白血病、小细胞肺癌、皮肤癌、上皮细胞癌、前列腺癌、非小细胞肺癌、鼻咽癌、恶性胶质瘤、淋巴瘤或黑色素瘤中的一种或多种。
说明书 :
一种喹唑啉酮与α，β-不饱和酮轭合衍生物及其制备方法和
应用
技术领域
[0001] 本发明属于药物化学技术领域，更具体地，涉及一种喹唑啉酮与α，β-不饱和酮轭合衍生物及其制备方法和应用。
背景技术
[0002] 恶性肿瘤是危害人类健康的一大类疾病，据世界卫生组织(WHO)统计，全世界每年有800万人死于癌症，而中国每年有接近200万人死于癌症。近年来抗恶性肿瘤药物的研发取得重大突破，许多新型药物进入临床研究，但仍存在诸多挑战。高毒副作用、耐药等问题仍然是临床肿瘤药物治疗面临的主要障碍。
[0003] c-MYC作为一个重要的原癌基因，调控着细胞增殖、分化、代谢、凋亡等生命过程。研究发现，c-MYC在多种恶性肿瘤中都有异常的表达，比如宫颈癌、乳腺癌、结肠癌、小细胞肺癌、恶性胶质瘤、黑色素瘤、骨肉瘤、骨髓行白血病等。大部分肿瘤细胞中高表达的c-MYC得益于基因的转录激活，c-MYC启动子核酸敏感元件NHEⅢ序列可形成G-四链体结构，并作为转录沉默子抑制c-MYC的转录。因此，靶向c-MYCG-四链体的配体的开发成为抗肿瘤药物研发的热点。
[0004] 喹唑啉基团在药物化学领域被广泛研究。喹唑啉类生物碱存在于多种植物和动物界中，而喹唑啉衍生物具有多种生物活性，包括抗肿瘤、抗菌、抗炎、降血压等功能。过去15年来，喹唑啉类衍生物在抗肿瘤方面取得重大进展，大量喹唑啉类化合物被报道，其中几个喹唑啉类化合物被FDA批准上市。
[0005] 查尔酮类化合物广泛存在于自然界中，其结构中含有α,β-不饱和酮、芳环，使其具有作为氢键受体、亲电性、亲酯性且整个分子表现出适当柔性，能与不同的受体结合，表现出多种生物活性。查尔酮类化合物对肿瘤发生、发展的多个阶段均有显著作用，包括抑制肿瘤细胞增殖、诱导肿瘤细胞分化和凋亡以及抑制肿瘤的发生和转移等。
发明内容
[0006] 本发明的目的之一在于提供一种喹唑啉酮与α，β-不饱和酮轭合衍生物。
[0007] 本发明的另一目的在于提供上述衍生物的制备方法。
[0008] 本发明的再一目的在于提供上述衍生物在作为原癌基因c-MYC转录调控阻断剂制备抗癌药物中的应用。
[0009] 本发明在喹唑啉酮结构基础上，与α,β-不饱和酮结构轭合得到新型的喹唑啉酮与α,β-不饱和酮轭合衍生物，该类衍生物具有抑制原癌基因c-myc转录和翻译的抗肿瘤作用。
[0010] 进一步地，本发明所提供的该类喹唑啉酮与α,β-不饱和酮轭合衍生物能够抑制原癌基因c-myc转录和翻译，抑制多株肿瘤细胞的增殖，具有广泛的抗肿瘤效果。
[0011] 本发明通过以下技术方案实现上述目的：
[0012] 本发明提供一种喹唑啉酮与α，β-不饱和酮轭合衍生物，所述喹唑啉酮与α，β-不饱和酮轭合衍生物的结构式如式(I)或式(II)所示：
[0013]
[0014] 其中，R1为氢、胺基、取代胺基、五元或六元杂环基；R2为氢、苯基、取代苯基、苯并杂环化合物；
[0015] 取代基选自C1-5烷基、C1-8烷氧基、卤基、苯基、胺基、五元或六元杂环基。
[0016] 优选地，R1为NH(CH2)n R3，R3为胺基、取代胺基、五元或六元杂环基，n为1～5中任意一个整数，取代胺基中的取代基选自C1-5烷基、C1-8烷氧基、五元或六元杂环基。
[0017] 优选地，R3为胺基、N,N-二乙基甲胺基、N,N-二甲基甲胺基或吗啉基。
[0018] 优选地，R2为苯基、取代苯基、苯并五元或苯并六元杂环化合物，取代苯基中的取代基选自卤基、C1-5烷氧基、五元或六元含氮杂环基。
[0019] 优选地，R2为苯基、取代苯基或1,3-苯并二噁茂，取代苯基中取代基为卤基、吗啉基、C1-5烷基或C1-5烷氧基。
[0020] 优选地，取代苯基中取代基为氟基、吗啉基、叔丁基或甲氧基。
[0021] 本发明同时提供所述的喹唑啉酮与α,β-不饱和酮轭合衍生物的制备方法，包括如下步骤：
[0022] S1.2-氨基-4,5-二氟苯甲酸与2-吡咯烷酮反应得到化合物
[0023] S2.化合物 与胺类化合物NH-R1反应得到中间体
[0024] S3.化合物 与酮类化合物反应得到中间体
[0025] S4.将S2中所得化合物与S3中所得化合物反应，得到所述衍生物。
[0026] 本发明同时提供所述的喹唑啉酮与α,β-不饱和酮轭合衍生物在制备抗癌药物中的应用。
[0027] 进一步地，所述喹唑啉酮与α,β-不饱和酮轭合衍生物是作为原癌基因c-MYC转录调控阻断剂应用于抗癌药物的制备。
[0028] 进一步地，所述抗癌为抗卵巢癌、宫颈癌、乳腺癌、肺腺癌、结肠癌、肝癌、白血病、小细胞肺癌、皮肤癌、上皮细胞癌、前列腺癌、非小细胞肺癌、鼻咽癌、恶性胶质瘤、淋巴瘤或黑色素瘤中的一种或多种。
[0029] 进一步地，本发明所述药物还包括和其药学上可接受的盐、载体、立体异构体或其前药分子。
[0030] 进一步地，本发明所述药物可以为注射剂、片剂、丸剂、胶囊剂、悬浮剂或乳剂。
[0031] 将本发明所提供的衍生物应用于抑制癌细胞实验，发现其具备显著的抑制效果，尤其是化合物3a、3c、3h、3i的抑制癌细胞株增殖效果非常显著。
[0032] 与现有技术相比，本发明具有以下优点：
[0033] 本发明涉及具有通式(I)或(II)结构特征的喹唑啉酮与α,β-不饱和酮轭合衍生物可以有效抑制多种肿瘤细胞的生长，并能够显著抑制原癌基因c-MYC的转录和表达，在制备抗肿瘤药物上有着广阔的应用空间。
附图说明
[0034] 图1为本发明提供的喹唑啉酮与α,β-不饱和酮轭合衍生物对c-MYC转录水平的影响图。
[0035] 图2为本发明提供的喹唑啉酮与α,β-不饱和酮轭合衍生物对C-MYC翻译水平的影响图。
具体实施方式
[0036] 下面结合具体实施例和附图进一步详细说明本发明。除非特别说明，本发明采用的试剂、设备和方法为本技术领域常规市购的试剂、设备和常规使用的方法。
[0037] 其中，本发明实施例14～33所述衍生物合成路线如下：
[0038]
[0039] 更具体地，本发明实施例14～33所述衍生物合成路线如下：
[0040] 2-氨基-4,5-二氟苯甲酸与2-吡咯烷酮反应得到化合物2 化合物2与不同的胺反应得到中间体3 和4
[0041] 化合物 与酮类化合物反应得到中间体ab c
d e
f g
h i
和j
d e
f g
h i
和j
[0042] 中间体3、4再与中间体a、b、c、d、e、f、g、h、i、j，溶于DMF后，在三甲基氯硅烷的作用下发生取代反应，得到最终化合物。在得到中间体
[0043] 中间体a、b、c、d、e、f、g、h、i、j的制备方法，使用对二苯甲醛或间二苯甲醛与苯乙酮类原料，在以NaOH为催化剂，以乙醇和水为反应溶剂的作用下发生反应，对二苯甲醛或间二苯甲醛与苯乙酮类原料反应的摩尔比为1：0.5～1，反应先置于冰浴中搅拌反应2h，再于室温搅拌反应2～6h。
[0044] 进一步地，终产物的制备方法，所述S4步骤中中间体3、4再与含取代基的苯甲醛中间体a、b、c、d、e、f、g、h、i、j反应的摩尔比为1：(1.05～3.0)，反应温度为50～120℃，反应时间为12～48小时。
[0045] 实施例1：化合物2的合成
[0046]
[0047] 将2-氨基-4,5-二氟苯甲酸(5g，29mmol)，2-吡咯烷酮(5mL)至于100mL单颈烧瓶中，于冰浴搅拌下逐滴加入20mL三氯氧磷(POCl3)，使其30min滴完，103℃加热回流24h，将反应体系逐滴加入200mL冰水中，用浓NaOH溶液调节pH至弱碱性，抽滤，得到粗产物，硅胶柱层析纯化(洗脱剂：V(石油醚)：V(乙酸乙酯)＝1:1)得到化合物m1，5.4g，产率85％。
[0048] 1H NMR(400MHz,CDCl3)δ8.20–7.85(m,1H),7.60–7.34(m,1H),4.42–4.06(m,2H),3.19(td,J＝7.9,2.7Hz,2H),2.43–2.16(m,2H).LC-MS m/z:223[M+H]+.
[0049] 实施例2：化合物3的合成
[0050]
[0051] 取化合物2(2.22g，10mmol)，二乙胺基丙胺(3ml，24mmol)，于厚壁耐压瓶中，100℃加热搅拌过夜，得到橙黄色溶液，反应体系降至室温后，有大量淡黄色沉淀析出，抽滤，得到滤饼为乳白色固体，抽真空干燥，得到乳白色固体2.3g，产率69％。
[0052] 1H NMR(400MHz,CDCl3)δ7.72(d,J＝11.7Hz,1H),6.72(s,1H),6.67(d,J＝7.7Hz,1H),4.19–4.12(m,2H),3.31(dd,J＝10.6,5.8Hz,2H),3.11(t,J＝7.9Hz,2H),2.63–2.59(m,2H),2.55(q,J＝7.1Hz,4H),2.30–2.19(m,2H),1.89–1.81(m,2H),1.06(t,J＝7.1Hz,
6H).
6H).
[0053] 实施例3：化合物4的合成
[0054]
[0055] 按照3的合成方法，得1.8g乳白色固体，产率52％。1H NMR(400MHz,CDCl3)δ7.67(d,J＝11.7Hz,1H),6.62(d,J＝7.7Hz,1H),4.13–4.05(m,2H),3.72–3.67(m,2H),3.66–3.61(m,2H),3.25(dd,J＝11.1,5.8Hz,2H),3.04(t,J＝7.9Hz,4H),2.51–2.46(m,3H),2.24–2.12(m,4H),1.86–1.75(m,2H).
[0056] 实施例4：化合物a的合成
[0057]
[0058] 称取对二苯甲醛(1.34g，10mol)，3′,4′-亚甲二氧苯乙酮(1.64g，10mol)与圆底烧瓶，加入3mL乙醇，1mL水，0.01g NaOH作为催化剂，置于冰浴中搅拌反应2h，再于室温搅拌反应2h。TLC监测反应结束后，抽滤，取滤饼，硅胶柱层析分离纯化，得淡黄色固体1.21g，产率43％。
[0059] 1H NMR(400MHz,DMSO)δ10.05(s,1H),8.12(d,J＝8.1Hz,2H),8.06(d,J＝11.5Hz,1H),7.97(d,J＝6.3Hz,2H),7.91(dd,J＝8.2,1.8Hz,1H),7.77(d,J＝9.3Hz,1H),7.69(s,
1H),7.11(d,J＝8.2Hz,1H),6.18(s,2H).
1H),7.11(d,J＝8.2Hz,1H),6.18(s,2H).
[0060] 实施例5：化合物b的合成
[0061]
[0062] 按照a的合成方法，得1.78g黄色固体，产率55％。
[0063] 1H NMR(400MHz,DMSO)δ10.05(s,1H),8.10(d,J＝7.5Hz,4H),8.07(s,1H),7.97(d,J＝8.1Hz,2H),7.72(d,J＝15.6Hz,1H),7.05(d,J＝8.9Hz,2H),3.77–3.72(m,4H),3.38–3.33(m,4H).
[0064] 实施例6：化合物c的合成
[0065]
[0066] 按照a的合成方法，得1.03g乳白色固体，产率35％。
[0067] 1H NMR(400MHz,DMSO)δ9.01(s,3H),8.12(d,J＝8.5Hz,1H),8.02(d,J＝15.7Hz,2H),7.99(m,2H),7.76(d,J＝15.6Hz,1H),7.60(d,J＝8.5Hz,1H),1.34(s,9H).
[0068] 实施例7：化合物d的合成
[0069]
[0070] 按照a的合成方法，得2.11g黄色固体，产率83％。
[0071] 1H NMR(400MHz,CDCl3)δ10.08(s,1H),8.10(ddd,J＝5.4,4.5,2.1Hz,2H),7.96(d,J＝8.2Hz,2H),7.83(d,J＝2.5Hz,2H),7.81(s,1H),7.68(d,J＝26.2Hz,1H),7.25–7.18(m,2H).
[0072] 实施例8：化合物e的合成
[0073]
[0074] 按照a的合成方法，得1.64g黄色固体，产率62％。
[0075] 1H NMR(400MHz,CDCl3)δ10.05(s,1H),8.06(d,J＝8.8Hz,2H),7.93(d,J＝8.1Hz,2H),7.83(d,J＝3.6Hz,1H),7.79(d,J＝7.8Hz,2H),7.65(d,J＝15.7Hz,1H),7.00(d,J＝
8.8Hz,2H),3.90(s,3H).
8.8Hz,2H),3.90(s,3H).
[0076] 实施例9：化合物f的合成
[0077]
[0078] 称取间二苯甲醛(1.34g，10mol)，3′,4′-亚甲二氧苯乙酮(1.64g，10mol)与圆底烧瓶，加入3mL乙醇，1mL水，0.01g NaOH作为催化剂，置于冰浴中搅拌反应2h，再于室温搅拌反应2h。TLC监测反应结束后，抽滤，取滤饼，硅胶柱层析分离纯化，得淡黄色固体1.01g，产率36％。
[0079] 1H NMR(400MHz,CDCl3)δ10.08(s,1H),8.15(s,1H),7.91(d,J＝7.6Hz,1H),7.86(d,J＝8.6Hz,1H),7.83(d,J＝15.9Hz,1H),7.68(dd,J＝8.1,1.4Hz,1H),7.63–7.53(m,3H),6.92(d,J＝8.1Hz,1H),6.08(s,2H).
[0080] 实施例10：化合物g的合成
[0081]
[0082] 按照f的合成方法，得1.22g黄色固体，产率38％。
[0083] 1H NMR(400MHz,CDCl3)δ10.10(s,1H),8.18(d,J＝5.5Hz,1H),8.11–8.01(m,2H),7.96–7.79(m,3H),7.73–7.58(m,2H),7.02–6.90(m,2H),3.95–3.85(m,4H),3.42–3.32(m,
4H).
4H).
[0084] 实施例11：化合物h的合成
[0085]
[0086] 按照f的合成方法，得1.20g乳白色固体，产率41％。
[0087] 1H NMR(400MHz,CDCl3)δ10.08(s,1H),8.17(s,1H),8.00(d,J＝8.4Hz,2H),7.94–7.89(m,2H),7.88(d,J＝7.4Hz,1H),7.62(d,J＝5.0Hz,1H),7.54(d,J＝8.3Hz,2H),7.48(dd,J＝8.5,2.3Hz,1H),1.37(s,9H).
[0088] 实施例12：化合物i的合成
[0089]
[0090] 按照f的合成方法，得1.68g黄色固体，产率66％。
[0091] 1H NMR(400MHz,CDCl3)δ10.09(s,1H),8.12–8.05(m,3H),7.86(d,J＝5.6Hz,1H),7.82(s,1H),7.70(dd,J＝7.7,1.5Hz,1H),7.56(d,J＝15.7Hz,1H),7.50(t,J＝7.7Hz,1H),
7.23–7.17(m,2H).
7.23–7.17(m,2H).
[0092] 实施例13：化合物j的合成
[0093]
[0094] 按照f的合成方法，得1.47g黄色固体，产率55％。
[0095] 1H NMR(400MHz,CDCl3)δ10.08(s,1H),8.16(s,1H),8.06(d,J＝8.8Hz,2H),7.89(dd,J＝16.9,7.2Hz,2H),7.85–7.79(m,1H),7.67(d,J＝8.5Hz,1H),7.63–7.60(m,1H),7.00(d,J＝8.6Hz,2H),3.91(s,3H).
[0096] 实施例14：化合物3a的合成
[0097]
[0098] 将化合物3(0.3323g，1mmol)与化合物a(0.3083g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.4180g，产率70％。
[0099] 1H NMR(400MHz,DMSO-d6)δ8.03(d,J＝6.8Hz,1H),8.02–7.97(m,J＝11.4,5.0Hz,3H),7.92–7.87(m,J＝8.2,3.8,1.7Hz,1H),7.76(d,J＝4.8Hz,1H),7.73–7.71(m,1H),
7.70–7.68(m,J＝4.9,2.4Hz,2H),7.63(d,J＝11.8Hz,1H),7.11(d,J＝8.2Hz,1H),6.93(d,J＝7.9Hz,1H),6.18(s,2H),4.20–4.12(m,2H),3.40–3.28(m,4H),3.20–3.06(m,J＝11.9,
13
5.9Hz,6H),2.09–1.95(m,2H),1.22(t,J＝7.2Hz,6H). C NMR(101MHz,DMSO-d6))δ187.46,
159.73,δ153.12(d,J＝194.2Hz),152.07,149.29,148.91,148.52,143.26(d,J＝13.7Hz),
142.94,137.82,135.45,135.05,132.84,130.42,129.72,127.98,125.54,123.26,109.54(d,J＝20.6Hz),109.11(d,J＝7.5Hz),108.60,108.41,106.01(d,J＝4.1Hz),102.54,+
51.09,46.96,44.33,41.72,29.42,25.79,24.96,11.55.ESI-HRMS[M+H] m/z＝595.2642,calcd for C35H35FN4O4,595.2699.Purity:99.6％by HPLC.
7.70–7.68(m,J＝4.9,2.4Hz,2H),7.63(d,J＝11.8Hz,1H),7.11(d,J＝8.2Hz,1H),6.93(d,J＝7.9Hz,1H),6.18(s,2H),4.20–4.12(m,2H),3.40–3.28(m,4H),3.20–3.06(m,J＝11.9,
13
5.9Hz,6H),2.09–1.95(m,2H),1.22(t,J＝7.2Hz,6H). C NMR(101MHz,DMSO-d6))δ187.46,
159.73,δ153.12(d,J＝194.2Hz),152.07,149.29,148.91,148.52,143.26(d,J＝13.7Hz),
142.94,137.82,135.45,135.05,132.84,130.42,129.72,127.98,125.54,123.26,109.54(d,J＝20.6Hz),109.11(d,J＝7.5Hz),108.60,108.41,106.01(d,J＝4.1Hz),102.54,+
51.09,46.96,44.33,41.72,29.42,25.79,24.96,11.55.ESI-HRMS[M+H] m/z＝595.2642,calcd for C35H35FN4O4,595.2699.Purity:99.6％by HPLC.
[0100] 实施例15：化合物3b的合成
[0101]
[0102] 将化合物3(0.3323g，1mmol)与化合物b(0.3535g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.2961g，产率46％。
[0103] 1H NMR(400MHz,CDCl3)δ8.02(d,J＝8.7Hz,2H),7.84–7.75(m,3H),7.71(d,J＝8.1Hz,2H),7.64–7.56(m,3H),6.93(d,J＝8.8Hz,2H),6.79(d,J＝7.6Hz,1H),4.28(t,J＝
7.0Hz,2H),3.91–3.84(m,4H),3.65(d,J＝4.5Hz,1H),3.41–3.27(m,8H),2.69–2.50(m,
13
6H),1.94–1.81(m,2H),1.08(t,J＝6.7Hz,6H). C NMR(101MHz,CDCl3)δ187.76,160.42,
154.96,154.26,150.87(d,J＝246.3Hz),148.71,143.46(d,J＝13.8Hz),142.16,137.38,
135.54,133.32,130.66,130.12,128.79,128.69,122.67,113.40,109.74(d,J＝20.8Hz),
109.42(d,J＝7.6Hz),105.55(d,J＝4.2Hz),70.59,66.57,52.72,47.49,46.85,43.78,
25.81,25.12,11.59.
7.0Hz,2H),3.91–3.84(m,4H),3.65(d,J＝4.5Hz,1H),3.41–3.27(m,8H),2.69–2.50(m,
13
6H),1.94–1.81(m,2H),1.08(t,J＝6.7Hz,6H). C NMR(101MHz,CDCl3)δ187.76,160.42,
154.96,154.26,150.87(d,J＝246.3Hz),148.71,143.46(d,J＝13.8Hz),142.16,137.38,
135.54,133.32,130.66,130.12,128.79,128.69,122.67,113.40,109.74(d,J＝20.8Hz),
109.42(d,J＝7.6Hz),105.55(d,J＝4.2Hz),70.59,66.57,52.72,47.49,46.85,43.78,
25.81,25.12,11.59.
[0104] ESI-HRMS[M+H]+m/z＝636.3272,calcd for C38H42FN5O3,636.3321.Purity:97.5％by HPLC.
[0105] 实施例16：化合物3c的合成
[0106]
[0107] 将化合物3(0.3323g，1mmol)与化合物c(0.3216g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.3050g，产率50％。
[0108] 1H NMR(400MHz,CDCl3)δ7.98(d,J＝8.0Hz,1H),7.81(d,J＝16.4Hz,1H),7.77–7.68(m,5H),7.59(d,J＝8.6Hz,2H),7.53(d,J＝7.6Hz,3H),6.78(d,J＝7.5Hz,1H),6.65(s,1H),4.30(t,J＝6.7Hz,2H),3.41–3.34(m,2H),3.33–3.24(m,2H),2.73–2.55(m,6H),
1.96–1.85(m,2H),1.37(s,9H),1.10(t,J＝6.6Hz,6H).
1.96–1.85(m,2H),1.37(s,9H),1.10(t,J＝6.6Hz,6H).
[0109] 13C NMR(101MHz,CDCl3)δ188.75,159.39,156.36(t,J＝273.9Hz),153.94,152.63,151.06,148.62,147.65,142.36(d,J＝14.1Hz),142.20,136.56,134.46,134.22,
132.46,131.30,129.88,129.13,127.81,127.79,127.72,127.49,124.63,121.79,108.76(d,J＝21.0Hz),108.47(d,J＝6.7Hz),104.53(d,J＝4.1Hz),64.54,51.46,45.80,42.89,
34.14,30.10,28.68,24.79,12.70.
132.46,131.30,129.88,129.13,127.81,127.79,127.72,127.49,124.63,121.79,108.76(d,J＝21.0Hz),108.47(d,J＝6.7Hz),104.53(d,J＝4.1Hz),64.54,51.46,45.80,42.89,
34.14,30.10,28.68,24.79,12.70.
[0110] ESI-HRMS[M+H]+m/z＝607.3370,calcd for C38H43FN4O2,607.3430.Purity:96.2％by HPLC.
[0111] 实施例17：化合物3d的合成
[0112]
[0113] 将化合物3(0.3323g，1mmol)与化合物d(0.2797g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.2507g，产率44％。
[0114] 1H NMR(400MHz,CDCl3)δ8.03–7.97(m,2H),7.71(m,3H),7.63(d,J＝8.3Hz,2H),7.52(d,J＝8.3Hz,2H),7.47(d,J＝15.6Hz,1H),7.12(t,J＝8.6Hz,2H),6.70(d,J＝7.7Hz,
1H),4.24–4.14(m,2H),3.29(dd,J＝10.6,5.6Hz,2H),3.25–3.18(m,2H),2.59(t,J＝
5.9Hz,2H),2.53(q,J＝7.0Hz,4H),1.84(dd,J＝11.7,5.8Hz,2H),1.02(t,J＝7.1Hz,6H).13C NMR(101MHz,CDCl3)δ187.50,165.94,163.40,159.40,153.87,149.86(d,J＝
245.0Hz),147.64,142.86,142.41(d,J＝14.0Hz),136.84,133.92,133.44,132.66,
130.13,130.04,129.14,127.85,127.58,121.16,114.91,114.69,108.74(d,J＝20.7Hz),
108.42(d,J＝7.7Hz),104.51(d,J＝3.7Hz),51.60,45.81,42.88,42.48,24.80,23.96,
10.44.
1H),4.24–4.14(m,2H),3.29(dd,J＝10.6,5.6Hz,2H),3.25–3.18(m,2H),2.59(t,J＝
5.9Hz,2H),2.53(q,J＝7.0Hz,4H),1.84(dd,J＝11.7,5.8Hz,2H),1.02(t,J＝7.1Hz,6H).13C NMR(101MHz,CDCl3)δ187.50,165.94,163.40,159.40,153.87,149.86(d,J＝
245.0Hz),147.64,142.86,142.41(d,J＝14.0Hz),136.84,133.92,133.44,132.66,
130.13,130.04,129.14,127.85,127.58,121.16,114.91,114.69,108.74(d,J＝20.7Hz),
108.42(d,J＝7.7Hz),104.51(d,J＝3.7Hz),51.60,45.81,42.88,42.48,24.80,23.96,
10.44.
[0115] ESI-HRMS[M+H]+m/z＝569.2650,calcd for C34H34F2N4O2,569.2722.Purity:98.2％by HPLC.
[0116] 实施例18：化合物3e的合成
[0117]
[0118] 将化合物3(0.3323g，1mmol)与化合物e(0.2930g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.3011g，产率52％。
[0119] 1H NMR(400MHz,CDCl3)δ8.05(d,J＝8.7Hz,2H),7.79(t,J＝14.6Hz,3H),7.71(d,J＝7.9Hz,2H),7.64–7.55(m,3H),7.00(d,J＝8.6Hz,2H),6.78(d,J＝7.6Hz,1H),4.27(t,J＝6.8Hz,2H),3.90(s,3H),3.36(m,2H),3.34–3.27(m,2H),2.70–2.53(m,J＝21.5Hz,6H),1.94–1.83(m,2H),1.09(t,J＝6.5Hz,6H).
[0120] 13C NMR(101MHz,CDCl3)δ188.28,163.53,160.37(d,J＝3.3Hz),153.47(d,J＝285.4Hz),149.62,148.66,143.45(d,J＝14.0Hz),142.77,137.50,135.22,133.44,
130.94,130.82,130.12,128.74,128.58,122.38,113.89,109.66(d,J＝20.8Hz),109.32(d,J＝7.7Hz),105.46(d,J＝4.2Hz),55.51,52.86,46.83,43.89,43.76,25.78,25.05,
11.67.
130.94,130.82,130.12,128.74,128.58,122.38,113.89,109.66(d,J＝20.8Hz),109.32(d,J＝7.7Hz),105.46(d,J＝4.2Hz),55.51,52.86,46.83,43.89,43.76,25.78,25.05,
11.67.
[0121] ESI-HRMS[M+H]+m/z＝581.2850,calcd for C35H37FN4O3,581.2900.Purity:99.1％by HPLC.
[0122] 实施例19：化合物3f的合成
[0123]
[0124] 将化合物3(0.3323g，1mmol)与化合物f(0.3083g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.2106g，产率35％。
[0125] 1H NMR(400MHz,CDCl3)δ7.82–7.77(m,1H),7.76(d,J＝3.4Hz,1H),7.73(d,J＝2.5Hz,1H),7.66(dd,J＝8.2,1.6Hz,1H),7.62(d,J＝7.5Hz,1H),7.57(d,J＝7.3Hz,1H),
7.55–7.53(m,1H),7.51–7.44(m,2H),6.91(dd,J＝8.1,1.5Hz,1H),6.77(d,J＝7.7Hz,2H),
6.07(d,J＝1.2Hz,2H),4.25(dd,J＝9.7,4.5Hz,2H),3.38–3.32(m,2H),3.28(dd,J＝8.6,
5.0Hz,2H),2.63(t,J＝5.7Hz,2H),2.57(q,J＝7.0Hz,4H),1.91–1.83(m,2H),1.07(t,J＝
7.1Hz,6H).13C NMR(101MHz,CDCl3)δ187.99,160.42,154.94,152.10–149.45(d,J＝
187.6Hz),149.61,148.40,143.45,143.31(d,J＝13.1Hz),136.44,135.66,133.16,
132.83,131.00,129.88,129.42,128.85,128.04,124.77,122.50,109.82(d,J＝20.8Hz),
109.57(d,J＝7.6Hz),108.44,107.97,105.61(d,J＝4.3Hz),101.94,52.18,46.75,43.91,
43.06,29.70,25.66,24.84,11.10.
7.55–7.53(m,1H),7.51–7.44(m,2H),6.91(dd,J＝8.1,1.5Hz,1H),6.77(d,J＝7.7Hz,2H),
6.07(d,J＝1.2Hz,2H),4.25(dd,J＝9.7,4.5Hz,2H),3.38–3.32(m,2H),3.28(dd,J＝8.6,
5.0Hz,2H),2.63(t,J＝5.7Hz,2H),2.57(q,J＝7.0Hz,4H),1.91–1.83(m,2H),1.07(t,J＝
7.1Hz,6H).13C NMR(101MHz,CDCl3)δ187.99,160.42,154.94,152.10–149.45(d,J＝
187.6Hz),149.61,148.40,143.45,143.31(d,J＝13.1Hz),136.44,135.66,133.16,
132.83,131.00,129.88,129.42,128.85,128.04,124.77,122.50,109.82(d,J＝20.8Hz),
109.57(d,J＝7.6Hz),108.44,107.97,105.61(d,J＝4.3Hz),101.94,52.18,46.75,43.91,
43.06,29.70,25.66,24.84,11.10.
[0126] ESI-HRMS[M+H]+m/z＝595.2642,calcd for C35H35FN4O4,595.2711.Purity:98.1％by HPLC.
[0127] 实施例20：化合物3g的合成
[0128]
[0129] 将化合物3(0.3323g，1mmol)与化合物g(0.3535g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.2360g，产率37％。
[0130] 1H NMR(400MHz,CDCl3)δ8.03(d,J＝9.0Hz,2H),7.85–7.76(m,4H),7.65(d,J＝7.7Hz,1H),7.63–7.57(m,2H),7.50(t,J＝7.7Hz,1H),6.94(d,J＝9.0Hz,2H),6.81(d,J＝
7.7Hz,1H),4.31–4.25(m,2H),3.90–3.86(m,4H),3.65(d,J＝4.5Hz,1H),3.42(t,J＝
5.4Hz,2H),3.38–3.34(m,4H),3.34–3.29(m,2H),2.83–2.68(m,6H),2.06–1.95(m,2H),
1.17(t,J＝6.7Hz,6H).13C NMR(101MHz,CDCl3)δ187.58,160.34,154.86,154.22,150.70(d,J＝244.5Hz),148.58,148.16,143.30(d,J＝14.2Hz),142.42,136.20,135.75,132.96,
130.80,130.64,129.81,129.31,128.68,128.39,128.00,122.37,113.30,109.60(d,J＝
20.5Hz),109.28(d,J＝7.6Hz),105.26(d,J＝35.6Hz),66.51,52.53,47.32,46.74,43.90,
43.42,25.56,24.85,11.41.
7.7Hz,1H),4.31–4.25(m,2H),3.90–3.86(m,4H),3.65(d,J＝4.5Hz,1H),3.42(t,J＝
5.4Hz,2H),3.38–3.34(m,4H),3.34–3.29(m,2H),2.83–2.68(m,6H),2.06–1.95(m,2H),
1.17(t,J＝6.7Hz,6H).13C NMR(101MHz,CDCl3)δ187.58,160.34,154.86,154.22,150.70(d,J＝244.5Hz),148.58,148.16,143.30(d,J＝14.2Hz),142.42,136.20,135.75,132.96,
130.80,130.64,129.81,129.31,128.68,128.39,128.00,122.37,113.30,109.60(d,J＝
20.5Hz),109.28(d,J＝7.6Hz),105.26(d,J＝35.6Hz),66.51,52.53,47.32,46.74,43.90,
43.42,25.56,24.85,11.41.
[0131] ESI-HRMS[M+H]+m/z＝636.3272,calcd for C38H42FN5O3,636.3317.Purity:98.1％by HPLC.
[0132] 实施例21：化合物3h的合成
[0133]
[0134] 将化合物3(0.3323g，1mmol)与化合物h(0.3216g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.1744g，产率29％。
[0135] 1H NMR(400MHz,CDCl3)δ8.12–8.03(m,2H),7.83(d,J＝15.7Hz,1H),7.80–7.72(m,3H),7.64(d,J＝7.4Hz,1H),7.60(d,J＝7.5Hz,1H),7.57–7.46(m,2H),7.20(t,J＝8.3Hz,
2H),6.85(s,1H),6.77(d,J＝7.6Hz,1H),4.27(t,J＝6.8Hz,2H),3.39–3.25(m,4H),2.62(t,J＝5.3Hz,2H),2.57(q,J＝7.2Hz,4H),1.93–1.81(m,2H),1.25(s,9H),1.07(t,J＝
7.1Hz,6H).13C NMR(101MHz,CDCl3)δ187.56,166.14,163.61,159.54,156.73(d,J＝
558.3Hz),151.18,148.75,147.79,143.37,142.62(d,J＝14.1Hz),135.57,134.50,
133.50,133.47,132.42,130.37,130.27,129.18,128.61,127.63,127.23,121.25,115.12,
114.90,108.80(d,J＝20.8Hz),108.46(d,J＝7.6Hz),104.60(d,J＝4.1Hz),52.00,50.24,
45.97,42.91,34.41,30.93,28.47,24.80,10.82.
2H),6.85(s,1H),6.77(d,J＝7.6Hz,1H),4.27(t,J＝6.8Hz,2H),3.39–3.25(m,4H),2.62(t,J＝5.3Hz,2H),2.57(q,J＝7.2Hz,4H),1.93–1.81(m,2H),1.25(s,9H),1.07(t,J＝
7.1Hz,6H).13C NMR(101MHz,CDCl3)δ187.56,166.14,163.61,159.54,156.73(d,J＝
558.3Hz),151.18,148.75,147.79,143.37,142.62(d,J＝14.1Hz),135.57,134.50,
133.50,133.47,132.42,130.37,130.27,129.18,128.61,127.63,127.23,121.25,115.12,
114.90,108.80(d,J＝20.8Hz),108.46(d,J＝7.6Hz),104.60(d,J＝4.1Hz),52.00,50.24,
45.97,42.91,34.41,30.93,28.47,24.80,10.82.
[0136] ESI-HRMS[M+H]+m/z＝607.3370,calcd for C38H43FN4O2,607.3462.Purity:98.6％by HPLC.
[0137] 实施例22：化合物3i的合成
[0138]
[0139] 将化合物3(0.3323g，1mmol)与化合物i(0.2797g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.3622g，产率64％。
[0140] 1H NMR(400MHz,CDCl3)δ8.12–8.04(m,2H),7.83(d,J＝15.7Hz,1H),7.80–7.72(m,3H),7.65(d,J＝7.6Hz,1H),7.61(d,J＝7.7Hz,1H),7.57–7.47(m,2H),7.20(t,J＝8.3Hz,
2H),6.85(s,1H),6.78(d,J＝7.6Hz,1H),4.27(t,J＝7.0Hz,2H),3.39–3.26(m,4H),2.63(t,J＝5.4Hz,2H),2.56(q,J＝7.0Hz,4H),1.93–1.83(m,2H),1.07(t,J＝7.0Hz,6H).13C NMR(101MHz,CDCl3)δ188.36,166.93,164.40,160.33,160.30,154.75,150.77(d,J＝
244.5Hz),148.58,144.17,143.41(d,J＝14.1Hz),136.37,135.30,134.30,134.27,
133.21,131.16,131.07,129.97,129.41,128.43,128.03,122.05,115.91,115.70,109.59(d,J＝20.8Hz),109.26(d,J＝7.6Hz),105.39(d,J＝4.1Hz),52.79,46.77,43.86,43.71,
29.77,25.59,25.00,11.62.
2H),6.85(s,1H),6.78(d,J＝7.6Hz,1H),4.27(t,J＝7.0Hz,2H),3.39–3.26(m,4H),2.63(t,J＝5.4Hz,2H),2.56(q,J＝7.0Hz,4H),1.93–1.83(m,2H),1.07(t,J＝7.0Hz,6H).13C NMR(101MHz,CDCl3)δ188.36,166.93,164.40,160.33,160.30,154.75,150.77(d,J＝
244.5Hz),148.58,144.17,143.41(d,J＝14.1Hz),136.37,135.30,134.30,134.27,
133.21,131.16,131.07,129.97,129.41,128.43,128.03,122.05,115.91,115.70,109.59(d,J＝20.8Hz),109.26(d,J＝7.6Hz),105.39(d,J＝4.1Hz),52.79,46.77,43.86,43.71,
29.77,25.59,25.00,11.62.
[0141] ESI-HRMS[M+H]+m/z＝569.2650,calcd for C34H34F2N4O2,569.2705.Purity:97.9％by HPLC.
[0142] 实施例23：化合物3j的合成
[0143]
[0144] 将化合物3(0.3323g，1mmol)与化合物j(0.2930g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.3140g，产率54％。
[0145] 1H NMR(400MHz,DMSO-d6)δ8.15(d,J＝8.5Hz,2H),8.03(s,1H),7.96–7.86(m,3H),7.75(d,J＝15.6Hz,1H),7.68(d,J＝7.8Hz,1H),7.61(d,J＝11.8Hz,1H),7.57(t,J＝
7.7Hz,1H),7.09(d,J＝8.5Hz,2H),7.01(d,J＝7.3Hz,1H),4.17(t,J＝6.9Hz,2H),3.88(s,
3H),3.41–3.32(m,4H),3.19–3.06(m,6H),2.13–2.03(m,2H),1.26(t,J＝7.2Hz,6H).13C NMR(101MHz,DMSO-d6)δ188.07,163.85,159.34(d,J＝3.1Hz),153.91(d,J＝427.6Hz),
149.36,147.15,146.21,143.17(d,J＝13.9Hz),142.91,136.42,136.13,133.83,131.48,
131.36,131.10,130.46,129.93,129.26,123.78,114.59,109.94(d,J＝20.8Hz),109.15(d,J＝7.4Hz),105.18(d,J＝3.9Hz),56.11,49.43,46.96,44.87,34.64,25.61,22.90,
8.95.
7.7Hz,1H),7.09(d,J＝8.5Hz,2H),7.01(d,J＝7.3Hz,1H),4.17(t,J＝6.9Hz,2H),3.88(s,
3H),3.41–3.32(m,4H),3.19–3.06(m,6H),2.13–2.03(m,2H),1.26(t,J＝7.2Hz,6H).13C NMR(101MHz,DMSO-d6)δ188.07,163.85,159.34(d,J＝3.1Hz),153.91(d,J＝427.6Hz),
149.36,147.15,146.21,143.17(d,J＝13.9Hz),142.91,136.42,136.13,133.83,131.48,
131.36,131.10,130.46,129.93,129.26,123.78,114.59,109.94(d,J＝20.8Hz),109.15(d,J＝7.4Hz),105.18(d,J＝3.9Hz),56.11,49.43,46.96,44.87,34.64,25.61,22.90,
8.95.
[0146] ESI-HRMS[M+H]+m/z＝581.2850,calcd for C35H37FN4O3,581.2904.Purity:97.1％by HPLC.
[0147] 实施例24：化合物4a的合成
[0148]
[0149] 将化合物4(0.3462g，1mmol)与化合物a(0.3083g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.2511g，产率41％。
[0150] 1H NMR(400MHz,CDCl3)δ7.82(d,J＝5.7Hz,1H),7.80–7.77(m,2H),7.71(d,J＝8.3Hz,2H),7.67(dd,J＝8.1,1.6Hz,1H),7.61(d,J＝8.3Hz,2H),7.55(dd,J＝8.6,7.0Hz,
2H),6.92(d,J＝8.1Hz,1H),6.82(d,J＝7.7Hz,1H),6.08(s,2H),4.32–4.26(m,2H),3.80(s,4H),3.39(d,J＝3.9Hz,2H),3.36–3.29(m,2H),2.56(d,J＝20.8Hz,6H),1.92(s,2H).13C NMR(101MHz,CDCl3)δ187.94,160.43(d,J＝3.5Hz),155.04,152.06,150.85(d,J＝
244.1Hz),148.68,148.39,143.36(d,J＝13.9Hz),143.08,137.57,135.25,133.44,
132.88,130.16,128.80,124.74,122.38,109.83(d,J＝20.9Hz),109.59(d,J＝7.7Hz),
108.43,107.97,105.63(d,J＝3.9Hz),101.94,66.89,58.10,53.78,43.93,43.45,25.81,
24.02.
2H),6.92(d,J＝8.1Hz,1H),6.82(d,J＝7.7Hz,1H),6.08(s,2H),4.32–4.26(m,2H),3.80(s,4H),3.39(d,J＝3.9Hz,2H),3.36–3.29(m,2H),2.56(d,J＝20.8Hz,6H),1.92(s,2H).13C NMR(101MHz,CDCl3)δ187.94,160.43(d,J＝3.5Hz),155.04,152.06,150.85(d,J＝
244.1Hz),148.68,148.39,143.36(d,J＝13.9Hz),143.08,137.57,135.25,133.44,
132.88,130.16,128.80,124.74,122.38,109.83(d,J＝20.9Hz),109.59(d,J＝7.7Hz),
108.43,107.97,105.63(d,J＝3.9Hz),101.94,66.89,58.10,53.78,43.93,43.45,25.81,
24.02.
[0151] ESI-HRMS[M+H]+m/z＝608.2435,calcd for C35H33FN4O5,609.2511.Purity:98.9％by HPLC.
[0152] 实施例25：化合物4b的合成
[0153]
[0154] 将化合物4(0.3462g，1mmol)与化合物b(0.3535g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.3100g，产率48％。
[0155] 1H NMR(400MHz,CDCl3)δ8.02(d,J＝8.8Hz,2H),7.80(dd,J＝11.5,4.2Hz,3H),7.71(d,J＝8.2Hz,2H),7.64–7.58(m,3H),6.93(d,J＝8.7Hz,2H),6.82(d,J＝7.7Hz,1H),
6.50(s,1H),4.28(t,J＝7.2Hz,2H),3.91–3.84(m,4H),3.79(s,4H),3.41–3.28(m,8H),
2.55(d,J＝22.9Hz,6H),1.91(s,2H).13C NMR(101MHz,CDCl3)δ187.74,160.42(d,J＝
3.2Hz),154.27,152.33(d,J＝545.4Hz),148.70,143.34(d,J＝13.9Hz),142.13,137.31,
135.58,133.25,130.66,130.13,128.85,128.69,122.68,113.40,109.81(d,J＝20.9Hz),
109.60(d,J＝7.6Hz),105.66(d,J＝3.9Hz),100.00,66.90,66.56,58.08,53.80,47.48,
43.92,43.42,25.79,24.09.
6.50(s,1H),4.28(t,J＝7.2Hz,2H),3.91–3.84(m,4H),3.79(s,4H),3.41–3.28(m,8H),
2.55(d,J＝22.9Hz,6H),1.91(s,2H).13C NMR(101MHz,CDCl3)δ187.74,160.42(d,J＝
3.2Hz),154.27,152.33(d,J＝545.4Hz),148.70,143.34(d,J＝13.9Hz),142.13,137.31,
135.58,133.25,130.66,130.13,128.85,128.69,122.68,113.40,109.81(d,J＝20.9Hz),
109.60(d,J＝7.6Hz),105.66(d,J＝3.9Hz),100.00,66.90,66.56,58.08,53.80,47.48,
43.92,43.42,25.79,24.09.
[0156] ESI-HRMS[M+H]+m/z＝650.3064,calcd forC38H40FN5O4,650.3145.Purity:99.4％by HPLC.
[0157] 实施例26：化合物4c的合成
[0158]
[0159] 将化合物4(0.3462g，1mmol)与化合物c(0.3216g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.1562g，产率25％。
[0160] 1H NMR(400MHz,CDCl3)δ7.99(d,J＝8.3Hz,2H),7.84(d,J＝5.8Hz,1H),7.80(d,J＝11.6Hz,3H),7.65(d,J＝7.5Hz,1H),7.60(t,J＝3.5Hz,2H),7.54(d,J＝8.7Hz,2H),7.53–7.47(m,1H),6.83(d,J＝7.7Hz,1H),6.50(s,1H),4.29(t,J＝7.1Hz,2H),3.87–3.73(m,4H),3.44–3.36(m,2H),3.35–3.27(m,2H),2.68–2.43(m,6H),1.98–1.84(m,2H),1.38(s,9H).13C NMR(101MHz,CDCl3)δ189.69,160.36(d,J＝3.2Hz),156.75,153.42(d,J＝
298.4Hz),149.51,148.56,143.50,143.26(d,J＝13.9Hz),136.29,135.56,135.30,
133.01,131.03,129.72,129.35,128.80,128.43,128.03,125.59,122.77,109.71(d,J＝
20.9Hz),109.46(d,J＝7.8Hz),105.52(d,J＝4.0Hz),66.79,57.99,53.67,43.84,43.34,
35.08,31.02,29.60,23.91.
298.4Hz),149.51,148.56,143.50,143.26(d,J＝13.9Hz),136.29,135.56,135.30,
133.01,131.03,129.72,129.35,128.80,128.43,128.03,125.59,122.77,109.71(d,J＝
20.9Hz),109.46(d,J＝7.8Hz),105.52(d,J＝4.0Hz),66.79,57.99,53.67,43.84,43.34,
35.08,31.02,29.60,23.91.
[0161] ESI-HRMS[M+H]+m/z＝621.3163,calcd for C38H41FN4O3,621.3209.Purity:99.0％by HPLC.
[0162] 实施例27：化合物4d的合成
[0163]
[0164] 将化合物4(0.3462g，1mmol)与化合物d(0.2797g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.2961g，产率51％。
[0165] 1H NMR(400MHz,CDCl3)δ8.10(dd,J＝7.8,5.8Hz,2H),7.88–7.79(m,3H),7.74(d,J＝8.1Hz,2H),7.63(d,J＝8.0Hz,2H),7.57(d,J＝15.6Hz,1H),7.22(t,J＝8.4Hz,2H),6.84(d,J＝7.6Hz,1H),6.54(s,1H),4.31(t,J＝7.0Hz,2H),3.89–3.76(m,4H),3.46–3.38(m,2H),3.37–3.30(m,2H),2.67–2.46(m,6H),1.99–1.89(m,2H).13C NMR(101MHz,CDCl3)δ
188.52,166.96,164.43,160.41(d,J＝3.4Hz),152.32(d,J＝537.2Hz),152.08,148.66,
143.87,143.37(d,J＝14.0Hz),137.82,134.43(d,J＝2.9Hz),134.42,133.64,131.16,
131.07,130.18,128.90,128.68,122.20,115.94,115.73,109.83(d,J＝21.0Hz),109.59(d,J＝7.8Hz),105.63(d,J＝4.1Hz),66.89,58.11,53.79,53.49,43.93,25.83,24.00.[0166] ESI-HRMS[M+H]+m/z＝583.2422,calcd for C34H32F2N4O3,583.2508.Purity:
98.9％by HPLC.
188.52,166.96,164.43,160.41(d,J＝3.4Hz),152.32(d,J＝537.2Hz),152.08,148.66,
143.87,143.37(d,J＝14.0Hz),137.82,134.43(d,J＝2.9Hz),134.42,133.64,131.16,
131.07,130.18,128.90,128.68,122.20,115.94,115.73,109.83(d,J＝21.0Hz),109.59(d,J＝7.8Hz),105.63(d,J＝4.1Hz),66.89,58.11,53.79,53.49,43.93,25.83,24.00.[0166] ESI-HRMS[M+H]+m/z＝583.2422,calcd for C34H32F2N4O3,583.2508.Purity:
98.9％by HPLC.
[0167] 实施例28：化合物4e的合成
[0168]
[0169] 将化合物4(0.3462g，1mmol)与化合物e(0.2930g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.3594g，产率60％。
[0170] 1H NMR(400MHz,DMSO)δ8.20(d,J＝8.2Hz,2H),8.07–7.99(m,3H),7.79–7.69(m,4H),7.64(d,J＝11.4Hz,1H),7.11(d,J＝7.9Hz,2H),6.90(d,J＝7.8Hz,1H),4.18–4.13(m,
2H),4.00–3.93(m,4H),3.88(s,3H),3.47–3.40(m,2H),3.34(dd,J＝14.4,7.1Hz,2H),
3.21–3.03(m,6H),2.10–2.02(m,2H).13C NMR(101MHz,DMSO)δ187.96,165.97,163.84,
159.58,151.97(d,J＝774.2Hz),149.33,143.01(d,J＝14.9Hz),142.60,137.64,135.73,
134.70,134.67,132.99,131.36,131.09,130.52,129.68,123.62,116.44,114.59,109.84(d,J＝19.9Hz),105.80(d,J＝6.7Hz),100.01(d,J＝2.6Hz),63.65,56.10,54.57,54.47,
51.61,44.61,25.81,22.66.
2H),4.00–3.93(m,4H),3.88(s,3H),3.47–3.40(m,2H),3.34(dd,J＝14.4,7.1Hz,2H),
3.21–3.03(m,6H),2.10–2.02(m,2H).13C NMR(101MHz,DMSO)δ187.96,165.97,163.84,
159.58,151.97(d,J＝774.2Hz),149.33,143.01(d,J＝14.9Hz),142.60,137.64,135.73,
134.70,134.67,132.99,131.36,131.09,130.52,129.68,123.62,116.44,114.59,109.84(d,J＝19.9Hz),105.80(d,J＝6.7Hz),100.01(d,J＝2.6Hz),63.65,56.10,54.57,54.47,
51.61,44.61,25.81,22.66.
[0171] ESI-HRMS[M+H]+m/z＝595.2642,calcd for C35H35FN4O4,595.2690.Purity:99.5％by HPLC.
[0172] 实施例29：化合物4f的合成
[0173]
[0174] 将化合物4(0.3462g，1mmol)与化合物f(0.3083g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.2530g，产率41％。
[0175] 1H NMR(400MHz,MeOD)δ8.13(s,1H),7.82–7.77(m,2H),7.73–7.66(m,3H),7.62(d,J＝3.8Hz,1H),7.60(d,J＝6.1Hz,1H),7.48(t,J＝7.8Hz,1H),7.42(s,1H),7.03(d,J＝7.3Hz,1H),6.86(d,J＝8.1Hz,1H),5.98(s,2H),4.33–4.28(m,2H),3.99(dd,J＝13.4,
2.0Hz,2H),3.72(t,J＝12.6Hz,2H),3.53–3.45(m,6H),3.41–3.34(m,4H),2.15–2.07(m,
2H).13C NMR(101MHz,MeOD)δ189.32,163.47,154.06(d,J＝303.1Hz),151.14,148.56,
143.18(d,J＝13.9Hz),142.78,141.42,138.63,131.72,130.86,129.57,128.57,125.13,
122.66,120.36,119.17,117.62,115.38,109.56(d,J＝14.6Hz),109.21(d,J＝8.9Hz),
107.63,102.15(d,J＝8.5Hz),63.70,56.93,54.92,51.99,30.25,25.32,24.71.
2.0Hz,2H),3.72(t,J＝12.6Hz,2H),3.53–3.45(m,6H),3.41–3.34(m,4H),2.15–2.07(m,
2H).13C NMR(101MHz,MeOD)δ189.32,163.47,154.06(d,J＝303.1Hz),151.14,148.56,
143.18(d,J＝13.9Hz),142.78,141.42,138.63,131.72,130.86,129.57,128.57,125.13,
122.66,120.36,119.17,117.62,115.38,109.56(d,J＝14.6Hz),109.21(d,J＝8.9Hz),
107.63,102.15(d,J＝8.5Hz),63.70,56.93,54.92,51.99,30.25,25.32,24.71.
[0176] ESI-HRMS[M+H]+m/z＝609.2435,calcd for C35H33FN4O5,609.2480.Purity:98.3％by HPLC.
[0177] 实施例30：化合物4g的合成
[0178]
[0179] 将化合物4(0.3462g，1mmol)与化合物g(0.3535g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.3961g，产率61％。
[0180] 1H NMR(400MHz,DMSO)δ8.07(d,J＝8.4Hz,2H),7.92(d,J＝16.1Hz,2H),7.84(d,J＝7.6Hz,1H),7.71(m,3H),7.60(d,J＝12.2Hz,1H),7.58–7.52(m,1H),7.02(d,J＝8.7Hz,2H),6.84(d,J＝7.9Hz,1H),6.59(s,1H),4.17–4.10(m,2H),3.78–3.72(m,4H),3.67–3.60(m,4H),3.37–3.28(m,8H),2.44–2.37(m,6H),1.85–1.77(m,2H).13C NMR(101MHz,CDCl3)δ
188.17,160.85(d,J＝3.1Hz),154.70,152.76(d,J＝542.4Hz),149.13,143.77(d,J＝
13.9Hz),142.56,137.74,136.01,133.68,131.09,130.56,129.28,129.12,123.11,
113.83,110.24(d,J＝20.6Hz),109.99(d,J＝7.4Hz),106.09(d,J＝3.9Hz),67.33,66.99,
58.51,54.23,47.91,44.35,43.85,26.22,24.52.
188.17,160.85(d,J＝3.1Hz),154.70,152.76(d,J＝542.4Hz),149.13,143.77(d,J＝
13.9Hz),142.56,137.74,136.01,133.68,131.09,130.56,129.28,129.12,123.11,
113.83,110.24(d,J＝20.6Hz),109.99(d,J＝7.4Hz),106.09(d,J＝3.9Hz),67.33,66.99,
58.51,54.23,47.91,44.35,43.85,26.22,24.52.
[0181] ESI-HRMS[M+H]+m/z＝650.3064,calcd for C38H40FN5O4,650.33119.Purity:99.5％by HPLC.
[0182] 实施例31：化合物4h的合成
[0183]
[0184] 将化合物4(0.3462g，1mmol)与化合物h(0.3216g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.1023g，产率16％。
[0185] 1H NMR(400MHz,CDCl3)δ8.02–7.98(m,2H),7.83–7.80(m,1H),7.78(d,J＝7.3Hz,2H),7.65(d,J＝7.6Hz,1H),7.59(d,J＝5.6Hz,1H),7.57–7.53(m,3H),7.50(t,J＝6.5Hz,
1H),6.82(d,J＝7.7Hz,1H),4.31–4.25(m,2H),3.89–3.77(m,4H),3.44–3.36(m,2H),3.32(dd,J＝9.9,4.3Hz,2H),2.69–2.47(m,6H),2.01–1.87(m,2H),1.37(s,9H).13C NMR(101MHz,CDCl3)δ189.80,160.47(d,J＝3.2Hz),156.86,153.53(d,J＝298.4Hz),149.62,
148.67,143.61,143.37(d,J＝13.9Hz),136.40,135.67,135.41,133.12,131.14,129.83,
129.46,128.91,128.54,128.14,125.70,122.88,109.82(d,J＝20.9Hz),109.57(d,J＝
7.8Hz),105.63(d,J＝4.0Hz),66.90,58.10,53.78,43.95,43.45,35.19,31.13,29.71,
25.68.
1H),6.82(d,J＝7.7Hz,1H),4.31–4.25(m,2H),3.89–3.77(m,4H),3.44–3.36(m,2H),3.32(dd,J＝9.9,4.3Hz,2H),2.69–2.47(m,6H),2.01–1.87(m,2H),1.37(s,9H).13C NMR(101MHz,CDCl3)δ189.80,160.47(d,J＝3.2Hz),156.86,153.53(d,J＝298.4Hz),149.62,
148.67,143.61,143.37(d,J＝13.9Hz),136.40,135.67,135.41,133.12,131.14,129.83,
129.46,128.91,128.54,128.14,125.70,122.88,109.82(d,J＝20.9Hz),109.57(d,J＝
7.8Hz),105.63(d,J＝4.0Hz),66.90,58.10,53.78,43.95,43.45,35.19,31.13,29.71,
25.68.
[0186] ESI-HRMS[M+H]+m/z＝621.3163,calcd for C38H41FN4O3,621.3220.Purity:97.9％by HPLC.
[0187] 实施例32：化合物4i的合成
[0188]
[0189] 将化合物4(0.3462g，1mmol)与化合物i(0.2797g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.4168g，产率72％。
[0190] 1H NMR(400MHz,CDCl3)δ8.08(t,J＝6.8Hz,2H),7.86(d,J＝12.0Hz,2H),7.80(d,J＝12.2Hz,2H),7.66(d,J＝7.8Hz,1H),7.61(d,J＝7.5Hz,1H),7.55(d,J＝11.5Hz,1H),7.53–7.49(m,1H),7.20(t,J＝8.4Hz,2H),6.83(d,J＝7.7Hz,1H),6.51(s,1H),4.29(t,J＝
7.0Hz,2H),3.87–3.73(m,4H),3.44–3.36(m,2H),3.35–3.29(m,2H),2.68–2.47(m,6H),
1.91(dt,J＝12.1,6.2Hz,2H).13C NMR(101MHz,CDCl3)δ188.49,166.98,164.45,160.40,
154.92,149.55,149.23(d,J＝548.0Hz),148.59,144.23,143.29(d,J＝14.0Hz),136.37,
135.35,134.29,133.16,131.20,131.11,130.04,129.49,128.69,128.10,122.16,115.97,
115.75,109.76(d,J＝21.0Hz),109.51(d,J＝8.0Hz),105.53(d,J＝3.4Hz),66.71,57.94,
53.65,43.94,43.26,25.62,23.85.
7.0Hz,2H),3.87–3.73(m,4H),3.44–3.36(m,2H),3.35–3.29(m,2H),2.68–2.47(m,6H),
1.91(dt,J＝12.1,6.2Hz,2H).13C NMR(101MHz,CDCl3)δ188.49,166.98,164.45,160.40,
154.92,149.55,149.23(d,J＝548.0Hz),148.59,144.23,143.29(d,J＝14.0Hz),136.37,
135.35,134.29,133.16,131.20,131.11,130.04,129.49,128.69,128.10,122.16,115.97,
115.75,109.76(d,J＝21.0Hz),109.51(d,J＝8.0Hz),105.53(d,J＝3.4Hz),66.71,57.94,
53.65,43.94,43.26,25.62,23.85.
[0191] ESI-HRMS[M+H]+m/z＝583.2442,calcd for C34H32F2N4O3,583.2500.Purity:99.1％by HPLC.
[0192] 实施例33：化合物4j的合成
[0193]
[0194] 将化合物4(0.3462g，1mmol)与化合物j(0.2930g，1.1mmol)加入后壁耐压瓶，再加入2mL的DMSO，10mL的三甲基氯硅烷，100℃加热过夜，TLC监测反应结束后，抽滤，粗产品用甲醇/二氯甲烷重结晶，得到淡黄色固体0.2010g，产率34％。
[0195] 1H NMR(400MHz,CDCl3)δ8.08(d,J＝8.8Hz,2H),7.85(d,J＝12.7Hz,1H),7.83–7.79(m,2H),7.68(d,J＝8.1Hz,1H),7.64–7.58(m,2H),7.56–7.50(m,1H),7.03(d,J＝
8.8Hz,2H),6.84(d,J＝7.7Hz,1H),4.35–4.27(m,2H),3.93(s,3H),3.90–3.84(m,4H),
3.47–3.40(m,2H),3.38–3.31(m,2H),2.80–2.44(m,6H),2.05–1.92(m,2H).13C NMR(101MHz,DMSO)δ187.86,163.83,159.29,154.86(d,J＝246.1Hz),151.34,147.84,143.21,
143.19(d,J＝3.6Hz),141.13,136.30,136.04,131.60,131.46,130.91,130.70,129.96,
129.39,123.42,118.74,114.55,111.77(d,J＝10.1Hz),109.92(d,J＝25.6Hz),104.63(d,J＝7.4Hz),63.61,56.09,54.43,51.51,45.00,34.59,25.61,22.41.
8.8Hz,2H),6.84(d,J＝7.7Hz,1H),4.35–4.27(m,2H),3.93(s,3H),3.90–3.84(m,4H),
3.47–3.40(m,2H),3.38–3.31(m,2H),2.80–2.44(m,6H),2.05–1.92(m,2H).13C NMR(101MHz,DMSO)δ187.86,163.83,159.29,154.86(d,J＝246.1Hz),151.34,147.84,143.21,
143.19(d,J＝3.6Hz),141.13,136.30,136.04,131.60,131.46,130.91,130.70,129.96,
129.39,123.42,118.74,114.55,111.77(d,J＝10.1Hz),109.92(d,J＝25.6Hz),104.63(d,J＝7.4Hz),63.61,56.09,54.43,51.51,45.00,34.59,25.61,22.41.
[0196] ESI-HRMS[M+H]+m/z＝595.2642,calcd for C35H35FN4O4,595.2692.Purity:98.7％by HPLC.
[0197] 实施例34：本发明所述的喹唑啉酮与α,β-不饱和酮轭合衍生物对肿瘤细胞生长的抑制作用
[0198] 选取本发明所述的喹唑啉酮与α,β-不饱和酮轭合衍生物，对多种肿瘤细胞株采用MTT法进行体外细胞毒测定。对数生长期的细胞加入不同浓度的喹唑啉酮与α,β-不饱和酮轭合衍生物，作用48h后，加入MTT，测定其吸光度。分别计算出抑制细胞生长50％时的化合物浓度，以IC50值表示，结果如表1所示。结果表明这系列化合物在体外对肿瘤细胞株有较强的抑制作用，可用于制备抗肿瘤药物。
[0199] 表1：喹唑啉酮与α,β-不饱和酮轭合衍生物对肿瘤细胞株生长的抑制作用(IC50值/μM)
[0200]
[0201] 实施例35：代表化合物3i对细胞内c-MYC基因的转录与表达的影响
[0202] RT-PCR实验方法：
[0203] 细胞培养：将细胞接种到6孔板中，200 000个/孔，待24小时细胞贴壁后，加入化合物，培养3-24小时后，收集细胞，提取RNA，检测RNA浓度，后按照下面体系做逆转录反应，及PCR实验。
[0204] 使用的PCR产物、引物、配制体系如下：
[0205] 10×Dream Taq Green buffer，2.5μL；dNTP mixture(2.5mM)，0.5μL；cDNA，1μL；引物(sense)，0.5μL；引物(antisense)，0.5μL；Dream TaqDNA聚合酶，0.25μL；ddH20，20.25μL；total，25μL
[0206] 程序:95℃变性，5min；95℃变性，30s；58℃退火，30s；72℃延伸，60s(30cycles)；72℃，10min；最终降温至10℃。
[0207] 1.5％琼脂糖电泳120V,0.5h，电泳结束后，凝胶成像。
[0208] 结果如图1所示，喹唑啉酮与α,β-不饱和酮轭合衍生物3i能够降低c-myc的转录水平。
[0209] Western blot实验方法：
[0210] 细胞培养：细胞记数，接种，培养在六孔板中，长至500万个细胞后，取出，裂解，细胞被收集后，加入50μL细胞裂解液，提取上清总蛋白液。使用BCA法检测总蛋白浓度，后变性蛋白样品，取相同质量的蛋白上样，SDS-PAGE胶电泳分离蛋白条带。根据目标蛋白计算分子量，将相应位置的电泳胶条带切下，湿转法将蛋白条带转到PVDF膜上。
[0211] 配置TBST缓冲液：25mM NaCl，100mM Tris,0.2％Tween-20,pH 7.4，用TBST缓冲液溶解的5％脱脂奶粉溶液(w/v)封闭PVDF膜。分别相应用一抗和二抗孵育PVDF膜，TBST缓冲液漂洗适当次数后，使用SuperECL Plus超敏发光试剂盒显色成像。
[0212] 结果如图2所示，喹唑啉酮与α,β-不饱和酮轭合衍生物3i能够降低C-MYC的表达水平。