含二氟甲基基团的3,3′‑螺环氧化吲哚及其制备方法和应用与流程

本发明涉及化学技术领域，尤其是一种含二氟甲基基团的3,3'-螺环氧化吲哚及其制备方法和应用。

背景技术：

二氟甲基(-cf2h)，作为一个重要的含氟基团，含有一个弱酸化的c-h键，是羟基、巯基及羟甲基的电子等排体，能够调节分子的生物活性、代谢稳定性以及亲脂性等，在医药、农药等领域起着与其他氟烷基同样重要的作用。因此，在分子中引入二氟甲基可赋予其诸多优良性能，同时引入或保持了生物靶点的关键识别元素。这些决定了其在生物医药、生物成像等领域的巨大潜力和科研价值。在另一方面，含有3,3'-螺环氧化吲哚骨架是许多临床药物分子和天然生物碱的核心结构单元，含有这种骨架的化合物往往具有良好的生物活性和潜在的药物价值。例如mi-888，具有很好的药代动力学性质以及很强的体内抗肿瘤活性；含有四氢硫吡喃的3,3'-螺环氧化吲哚是一种新型的p53-mdm2抑制剂。鉴于3,3'-螺环氧化吲哚具有潜在的生物活性以及二氟甲基在药物化学中的重要作用，因此，以二氟甲基重氮甲烷为砌块，通过[3+2]成环策略与3-烯基氧化吲哚反应，合成含有二氟甲基基团的3,3'-螺环氧化吲哚可能会产生一系列结构和活性上有意义的新化合物，它们的合成可以为二氟甲基取代的3,3'-螺环氧化吲哚在药物发现和成药性评价中的应用建立物质基础。

技术实现要素：

本发明的目的在于提供一种含二氟甲基基团的3,3'-螺环氧化吲哚及其制备方法和应用，该类化合物是一类重要的药物活性分子，对药物筛选和制药行业具有重要的应用价值，并且该类分子的合成具有反应条件温和、操作简单、底物普适性好的优点。

本发明是这样实现的：原位生成的二氟甲基重氮甲烷与靛红衍生的3-烯基氧化吲哚在无催化剂条件下，在有机溶剂中进行[3+2]反应，获得含二氟甲基基团的3,3'-螺环氧化吲哚。该类化合物具有如通式(ⅰ)所示的结构：

式中，r¹为烷基、烷氧基或卤素；r²为烷基；r³为烷基、芳基或酰基。

所述的有机溶剂为二氯甲烷、三氯甲烷、四氯化碳、乙腈、四氢呋喃、苯、甲苯、二甲苯、均三甲苯、1,4-二氧六环、乙二醇二甲醚、乙二醇二乙醚或甲基叔丁基醚。

原位生成的二氟甲基重氮甲烷与靛红衍生的3-烯基氧化吲哚的反应温度为25-100℃，反应时间为1-48小时。

所述的含二氟甲基基团的3,3'-螺环氧化吲哚在制备防治肿瘤疾病药物的应用。

本发明的反应原理如下：

其中，r¹，r²，r³和有机溶剂如上所述。

通过采用上述技术方案，二氟乙胺(1)放置于反应瓶中，依次加入有机溶剂、亚硝酸叔丁酯和乙酸，并在70℃条件下反应10分钟，停止加热，即可原位得到二氟甲基重氮甲烷(2)。然后将靛红衍生的3-烯基氧化吲哚(3)加入反应瓶中，经过[3+2]成环反应，合成了一系列含二氟甲基基团的3,3'-螺环氧化吲哚(4)。该类化合物含有潜在生物活性的二氟甲基基团和3,3'-螺环氧化吲哚骨架，可以为药物发现和成药性评价建立物质基础，具有重要的应用价值。本发明原料易于获得，可以在各种有机溶剂中进行，具有反应条件温和、操作简单、底物普适性好的优点。

附图说明

图1为本发明的实施例化合物4a的单晶-x射线衍射结构。

图2为本发明的实施例化合物4k的单晶-x射线衍射结构。

图3为本发明的实施例化合物4t的单晶-x射线衍射结构。

图4-6为本发明的实施例化合物4a的核磁共振谱图。

具体实施方式

本发明的实施例1：含二氟甲基基团的3,3'-螺环氧化吲哚(4)的制备；化合物4a：在圆底烧瓶中，依次加入二氯甲烷(15ml)，二氟乙胺(1,97.3mg,1.2mmol)，亚硝酸叔丁酯(148.5mg,1.44mmol)，乙酸(14.5mg,0.24mmol)，在70℃条件下反应10分钟，停止加热，即可原位制备获得二氟甲基重氮甲烷(2)。再加入靛红衍生的3-烯基氧化吲哚(3a，92.5mg,0.4mmol)，并在室温下充分搅拌24小时，薄层色谱检测反应完毕后，经减压浓缩、硅胶柱层析(300-400目)分离(石油醚：乙酸乙酯＝5：1)，得到白色固体108.6mg，产率84％，熔点：91.8–93.8℃，核磁共振和高分辨质谱测试分析结果如下：99:1dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ7.39(t,j＝7.8hz,1h),7.00(t,j＝7.6hz,1h),6.94(d,j＝7.8hz,1h),6.80(d,j＝7.6hz,1h),6.44(td,j＝4.0,54.4hz,1h),5.82-5.73(m,1h),3.90-3.73(m,2h),3.57(d,j＝8.4hz,1h),3.32(s,3h),0.68(t,j＝7.2hz,3h)；¹³cnmr(100mhz,cdcl3)δ171.0,167.9,144.7,131.5,124.9,123.4,121.6,112.9(t,j＝244.0hz),109.0,99.3,94.0(t,j＝22.0hz),61.9,44.3(t,j＝3.0hz),27.2,13.5；¹⁹fnmr(376mhz,cdcl3)δ-122.74(abdd,j＝8.9,54.3,294.8hz),-126.64(abdd,j＝16.2,55.4,294.6hz)；hrms(esi-tof)calcd.forc15h15f2n3nao3[m+na]⁺:346.0974；found:346.0971.

附图中单晶-x射线衍射结构图和核磁共振谱图由现有技术软件生成，本领域技术人员可以通过图中的特点即可了解各化合物属性，对于图中的数字可能因提交原因不是很清晰，但不影响本发明的完全公开。

通过实施例制备的化合物4b～4u的制备方法同化合物4a，投料比与化合物4a相同，可得到化合物4b～4u，反应产率见表1，但需强调的是本发明的化合物不限于表1所表示的内容。

表1为含二氟甲基基团的3,3'-螺环氧化吲哚的化学结构

表1：

本实施例制备化合物4b：黄色油状物，产率72％；核磁共振和高分辨质谱测试分析结果如下：95:5dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ(major+minor)7.19(d,j＝7.8hz,0.95h),7.14(s,0.05h),7.11(d,j＝8.0hz,0.05h),6.82(d,j＝7.8hz,0.95h),6.78(d,j＝8.0hz,0.05h),6.60(s,0.95h),6.43(td,j＝4.0,55.5hz,0.95h),6.38(td,j＝7.0,54.5hz,0.05h),5.83-5.73(m,1h),4.23-4.08(m,0.10h),3.92-3.73(m,1.90h),3.56(d,j＝8.2hz,1h),3.32(s,2.85h),3.24(s,0.15h),2.31(s,0.15h),2.24(s,2.85h),1.21(t,j＝7.2hz,0.15h),0.70(t,j＝7.0hz,2.85h)；¹³cnmr(100mhz,cdcl3)δ(major+minor)170.9,167.9,142.2,133.1,132.4,131.6,128.9,125.5,123.6,121.5,113.0(t,j＝243.6hz),108.8,108.3,99.4,93.8(t,j＝22.0hz),61.9,44.3(t,j＝2.9hz),27.2,26.9,21.3,21.0,14.2,13.5；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.04(abdd,j＝5.8,54.4,293.6hz,minor),-115.85(abdd,j＝6.8,55.8,294.5hz,minor),-121.63(abdd,j＝9.4,54.4,294.7hz,major),-125.51(abdd,j＝16.1,55.3,294.7hz,major)；hrms(esi-tof)calcd.forc16h17f2n3nao4[m+na]⁺:360.1130；found:360.1147.

本实施例制备化合物4c：红色油状物，产率77％；核磁共振和高分辨质谱测试分析结果如下：85:15dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ(major+minor)6.93(d,j＝2.5hz,0.15h),6.90(d,j＝2.5hz,0.85h),6.86(s,0.85h),6.83(s,0.15h),6.42(td,j＝4.1,55.0hz,1h),6.40(d,j＝2.5hz,1h),5.83-5.74(m,1h),3.91-3.78(m,2h),3.76(s,0.45h),3.70(s,2.55h),3.56(d,j＝8.2hz,1h),3.31(s,2.55h),3.23(s,0.45h),1.21(t,j＝7.1hz,0.45h),0.74(t,j＝7.1hz,2.55h)；¹³cnmr(100mhz,cdcl3)δ(major+minor)170.6,167.9,156.3,156.1,137.9,125.2,122.5,116.0,113.4,113.0(t,j＝243.6hz),112.0,110.0,109.5,108.9,99.7,94.0(t,j＝22.0hz),61.9,56.0,44.3(t,j＝2.8hz),27.2,26.9,14.1,13.6；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.06(abdd,j＝6.0,55.0,294.4hz,minor),-115.87(abdd,j＝7.2,55.8,294.3hz,minor),-121.65(abdd,j＝9.2,54.4,294.8hz,major),-125.45(abdd,j＝15.9,55.2,294.7hz,major)；hrms(esi-tof)calcd.forc16h17f2n3nao4[m+na]⁺:376.1079；found:376.1085.

本实施例制备化合物4d：黄色固体，产率77％；熔点:86.7–88.6℃；核磁共振和高分辨质谱测试分析结果如下：84:16dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ7.13(t,j＝8.6hz,1h),6.90(dd,j＝3.5,8.4hz,1h),6.59(s,1h),6.45(td,j＝4.0,55.0,1h),5.86-5.77(m,1h),3.97-3.80(m,2h),3.58(d,j＝8.0hz,1h),3.34(s,3h),0.79(t,j＝7.1hz,3h)；¹³cnmr(100mhz,cdcl3)δ170.6,167.7,159.2(d,j＝242.3hz),140.7,122.8(d,j＝8.2hz),117.8(d,j＝23.3hz),113.3(d,j＝25.5hz),112.8(t,j＝243.8hz),109.7(d,j＝8.0hz),99.4,94.5(t,j＝22.3hz),62.2,44.2(t,j＝2.8hz),27.4,13.7；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.09(abdd,j＝6.0,54.6,295.4hz,minor),-116.01(abdd,j＝7.5,55.7,295.3hz,minor),(-118.52)–(-118.58)(m,major),(-119.44)–(-119.50)(m,minor),-121.61(abdd,j＝9.0,54.4,295.2hz,major),-125.52(abdd,j＝16.2,55.3,295.3hz,major)；hrms(esi-tof)calcd.forc15h14f3n3nao3[m+na]⁺:364.0879；found:364.0870.

本实施例制备化合物4e：红色油状物，产率74％；核磁共振和高分辨质谱测试分析结果如下：87:13dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ(major+minor)7.39(dd,j＝2.0,8.2hz,0.87h),7.36(d,j＝2.0hz,0.13h),7.29(dd,j＝2.0,8.2hz,0.13h),6.89(d,j＝8.2hz,0.87h),6.82(s,0.13h),6.80(d,j＝2.0,0.87h),6.45(td,j＝4.0,55.0hz,0.87h),6.36(td,j＝7.0,55.2hz,0.13h),5.86-5.77(m,1h),4.24-4.11(m,0.26h),3.96-3.81(m,1.74h),3.57(d,j＝7.6hz,1h),3.33(s,2.61h),3.25(s,0.39h),1.23(t,j＝7.0hz,0.39),0.78(t,j＝7.0hz,2.61h)；¹³cnmr(100mhz,cdcl3)δ(major+minor)170.5,167.7,143.3,131.4,128.8,128.6,128.3,125.4,123.5,123.1,112.8(t,j＝243.8hz),110.0,109.4,99.1,94.6(t,j＝22.2hz),62.2,44.2(t,j＝2.9hz),27.3,27.0,14.2,13.7；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.06(abdd,j＝5.5,55.8,296.0hz,minor),-116.00(abdd,j＝7.1,55.5,295.4hz,minor),-121.69(abdd,j＝8.9,54.2,295.2hz,major),-125.52(abdd,j＝16.3,55.2,295.0hz,major)；hrms(esi-tof)calcd.forc15h15clf2n3o3[m+h]⁺:358.0765；found:358.0774.

本实施例制备化合物4f：橘红色油状物，产率72％；核磁共振和高分辨质谱测试分析结果如下：81:19dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ(major+minor)7.54(dd,j＝2.0,8.4hz,0.81h),7.49(d,j＝2.0hz,0.19h),7.44(dd,j＝2.0,8.4hz,0.19h),6.93(d,j＝2.0hz,0.81h),6.84(d,j＝8.4hz,0.81h),6.77(d,j＝8.4hz,0.19h),6.45(td,j＝4.0,54.8hz,0.81h),6.36(td,j＝7.2,55.2hz,0.19h),5.84-5.77(m,1h),3.99-3.91(m,1h),3.88-3.80(m,1h),3.57(d,j＝8.0hz,1h),3.33(s,2.43h),3.25(s,0.57h),1.25(td,j＝1.5,7.2hz,0.57h),0.80(t,j＝7.2hz,2.43h)；¹³cnmr(100mhz,cdcl3)δ(major+minor)170.4,167.6,143.7,134.3,131.5,128.1,126.2,123.4,115.8,112.8(t,j＝243.9hz),110.5,109.9,99.0,94.5(t,j＝22.2hz),62.3,60.5,44.2(t,j＝2.8hz),27.3,27.0,21.2,14.3(d,j＝11.5hz),13.7；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.05(abdd,j＝6.0,54.6,294.9hz,minor),-116.00(abdd,j＝7.2,55.7,295.5hz,minor),-121.71(abdd,j＝9.0,54.3,295.2hz,major),-125.50(abdd,j＝16.3,55.3,295.3hz,major)；hrms(esi-tof)calcd.forc15h14brf2n3nao3[m+na]⁺:424.0079；found:424.0070.

本实施例制备化合物4g：黄色油状物，产率59％；核磁共振和高分辨质谱测试分析结果如下：90:10dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ(major+minor)7.72(dd,j＝2.0,8.2hz,1.0h),7.63(s,0.10h),7.07(s,0.90h),6.73(d,j＝8.2hz,0.90h),6.67(d,j＝8.2hz,0.10h),6.44(td,j＝3.2hz,0.90h),6.35(td,j＝7.6,55.0hz,0.10h),5.83-5.78(m,1.0h),3.99-3.78(m,2h),3.55(d,j＝8.0hz,0.90h),3.46(d,j＝7.2hz,0.10h),3.31(s,2.7h),3.23(s,0.3h),1.19(t,j＝7.2hz,0.30h),0.78(t,j＝7.2hz,2.7h)；¹³cnmr(100mhz,cdcl3)δ(major+minor)170.2,167.6,144.3,140.2,137.5,133.4,131.6,123.7,112.8(t,j＝243.8hz),111.0,110.5,98.8,94.4(t,j＝22.2hz),85.2,62.2,44.2(t,j＝2.8hz),27.2,26.9,14.2,13.7；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.00(abdd,j＝6.4,55.0,295.5hz,minor),-115.94(abdd,j＝7.8,55.8,295.5hz,minor),-121.74(abdd,j＝9.3,54.3,294.6hz,major),-125.44(abdd,j＝16.4,55.2,294.9hz,major)；hrms(esi-tof)calcd.forc15h14if2n3nao3[m+na]⁺:471.9940；found:471.9970.

本实施例制备化合物4h：黄色油状物，产率70％；核磁共振和高分辨质谱测试分析结果如下：76:24dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ(major+minor)7.31(d,j＝8.6hz,0.76h),7.28(s,0.24h),7.20(d,j＝8.4hz,0.24),6.96(d,j＝8.6hz,0.76h),6.88(d,j＝8.4hz,0.24h),6.73(s,0.76h),6.45(td,j＝4.0,54.8hz,0.76h),6.38(td,j＝7.2,55.1hz,0.24h),5.89-5.80(m,1h),4.21-4.11(m,0.48h),3.94-3.78(m,1.52h),3.58(d,j＝8.0hz,1h),3.36(s,2.28h),3.28(s,0.72h),1.22(t,j＝7.2hz,0.72h),0.77(t,j＝7.2hz,2.28h)；¹³cnmr(100mhz,cdcl3)δ(major+minor)172.1,170.7,167.6,166.5,144.9,144.8,143.4,142.9,125.5,124.7,122.9,121.9,120.5(q,j＝256.0hz),119.2,117.3,113.1(t,j＝236.0hz),112.8(t,j＝243.8hz),109.6,108.9,99.2,94.8(t,j＝22.2hz),62.3,62.2,44.2(t,j＝2.9hz),27.4,27.0,14.1,13.5；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-58.49(s,minor),-58.61(s,major),-111.10(abdd,j＝5.7,54.7,295.5hz,minor),-116.08(abdd,j＝7.1,55.6,295.5hz,minor),-121.79(abdd,j＝8.9,54.2,295.2hz,major),-125.53(abdd,j＝16.4,55.5,295.4hz,major)；hrms(esi-tof)calcd.forc16h14f5n3nao4[m+na]⁺:430.0797；found:430.0795.

本实施例制备化合物4i：黄色固体，产率58％；熔点:75.4–77.4℃；核磁共振和高分辨质谱测试分析结果如下：93:7dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ6.99(dd,j＝1.9,8.0hz,1h),6.96(s,1h),6.72(d,j＝8.0hz,1h),6.45(td,j＝3.9,54.9hz,1h),5.83-5.73(m,1h),3.92-3.78(m,2h),3.56(d,j＝10.6hz,1h),3.33(s,3h),0.78(t,j＝7.1hz,3h)；¹³cnmr(100mhz,cdcl3)δ170.9,167.7,145.8,137.5,125.8,123.2,119.8,112.7(t,j＝243.6hz),109.8,98.8,94.2(t,j＝22.2hz),62.1,44.1(t,j＝2.8hz),27.3,13.6；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.01(abdd,j＝5.8,54.6,294.8hz,minor),-115.92(abdd,j＝7.0,55.7,294.9hz,minor),-121.92(abdd,j＝8.9,54.3,294.8hz,major),-125.68(abdd,j＝16.5,55.6,294.8hz,major)；hrms(esi-tof)calcd.forc15h14clf2n3nao3[m+na]⁺:380.0584；found:380.0573.

本实施例制备化合物4j：红色固体，产率53％；熔点:82.6–84.2℃；核磁共振和高分辨质谱测试分析结果如下：72:28dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ(major+minor)7.21(dd,j＝1.2,8.2hz,0.28h),7.16(d,j＝8.0hz,0.28h),7.15(d,j＝1.8,8.0hz,0.72h),7.11(d,j＝1.5hz,0.72h),7.04(d,j＝1.6hz,0.28h),6.66(dd,j＝1.2,8.0hz,0.72h),6.45(td,j＝4.0,55.4hz,0.72h),6.36(td,j＝7.2,55.2hz,0.28h),5.83-5.73(m,1h),4.20-4.07(m,0.56h),3.90-3.80(m,1.44h),3.56(dd,j＝1.2,8.2hz,0.72h),3.32(d,j＝1.4hz,2.16h),3.24(d,j＝1.4hz,0.84h),3.23(dd,j＝1.4,9.4hz,0.28h),1.21(td,j＝1.5,7.2hz,0.84h),0.79(td,j＝1.5,7.2hz,2.16h)；¹³cnmr(100mhz,cdcl3)δ(major+minor)172.1,170.8,167.7,166.7(d,j＝20.7hz),145.9,145.4,126.1(d,j＝3.5hz),125.6,125.4,124.2,122.9,122.4,120.4,115.5,113.1,112.7(t,j＝243.7hz),112.6,112.1,110.7,98.9,94.3(t,j＝22.1hz),62.1,44.1(t,j＝2.8hz),27.3,27.0,14.1,13.6；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.00(abdd,j＝6.3,54.6,295.0hz,minor),-115.93(abdd,j＝7.3,55.6,295.2hz,minor),-121.85(abdd,j＝8.5,54.3,295.0hz,major),-125.68(abdd,j＝16.4,55.2,294.8hz,major)；hrms(esi-tof)calcd.forc15h14brf2n3nao3[m+na]⁺:424.0079；found:424.0083.

本实施例制备化合物4k：橘红色固体，产率65％；熔点:98.9–100.9℃；核磁共振和高分辨质谱测试分析结果如下：97:3dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ6.97-6.92(m,2h),6.42(td,j＝4.1,54.9hz,1h),6.37(dd,j＝2.2,6.3hz,1h),5.80-5.71(m,1h),3.91-3.77(m,2h),3.88(s,3h),3.60(s,3h),3.56(d,j＝8.4hz,1h),0.75(t,j＝7.2hz,3h)；¹³cnmr(100mhz,cdcl3)δ171.1,167.8,145.8,132.4,124.0,122.8,117.2,115.1,113.0(t,j＝243.6hz),99.4,93.8(t,j＝22.0hz),61.9,56.3,44.5(t,j＝2.8hz),30.6,13.5；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.11(abd,j＝54.5,294.2hz,minor),-116.01(abd,j＝54.8,310.9hz,minor),-121.75(abdd,j＝9.5,54.4,294.4hz,major),-125.53(abdd,j＝15.9,55.3,294.5hz,major)；hrms(esi-tof)calcd.forc16h17f2n3nao4[m+na]⁺:376.1079；found:376.1079.

本实施例制备化合物4l：黄色固体，产率64％；熔点:85.3–87.1℃；核磁共振和高分辨质谱测试分析结果如下：91:9dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ(major+minor)7.18-7.12(m,1h),7.00-6.94(m,1h),6.60-6.57(m,1h),6.46(td,j＝3.9,55.0hz,1h),5.85-5.74(m,1h),3.93-3.78(m,2h),3.61-3.53(m,4h),0.78(td,j＝1.2,7.2hz,3h)；¹³cnmr(100mhz,cdcl3)δ(major+minor)170.6,167.7,147.9(d,j＝244.2hz),131.4(d,j＝8.9hz),124.2(d,j＝3.2hz),124.1(d,j＝6.4hz),121.4,120.8(d,j＝3.4hz),119.5(d,j＝19.0hz),112.8(t,j＝243.8hz),99.2,94.2(t,j＝22.2hz),65.1,62.1,46.6,44.5(t,j＝2.8hz),29.8(d,j＝5.8hz),13.8,13.7；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.08(abdd,j＝5.3,53.7,296.0hz,minor),-116.12(abdd,j＝7.0,55.5,295.0hz,minor),-121.83(abdd,j＝8.9,54.0,295.0hz,major),-125.70(abdd,j＝16.4,55.2,294.9hz,major),(-135.17)–(-135.23)(m,major),(-135.43)–(-135.48)(m,minor)；hrms(esi-tof)calcd.forc15h14f3n3nao3[m+na]⁺:364.0879；found:364.0878.

本实施例制备化合物4m：黄色固体，产率67％；熔点:77.4–79.2℃；核磁共振和高分辨质谱测试分析结果如下：94:6dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ7.33(d,j＝8.2hz,1h),6,93(t,j＝7.6hz,1h),6.64(j＝7.4hz,1h),6.46(td,j＝3.2,54.4hz,1h),5.81-5.73(m,1h),3.93-3.78(m,2h),3.71(s,3h),3.59(d,j＝8.2hz,1h),0.77(t,j＝7.1hz,3h)；¹³cnmr(100mhz,cdcl3)δ171.3,167.6,140.5,133.6,131.0,124.1,123.3,116.5,112.7(t,j＝243.7hz),98.7,94.1(t,j＝22.2hz),62.1,44.6(t,j＝2.8hz),30.7,13.6；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.07(abdd,j＝5.8,54.5,295.0hz,minor),-114.68(abdd,j＝7.0,55.0,292.6hz,minor),-122.00(abdd,j＝9.0,54.1,294.7hz,major),-125.77(abdd,j＝16.4,55.2,294.9hz,major)；hrms(esi-tof)calcd.forc15h14clf2n3nao3[m+na]⁺:380.0584；found:380.0579.

本实施例制备化合物4n：黄色固体，产率60％；熔点:91.4–93.2℃；核磁共振和高分辨质谱测试分析结果如下：93:7dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ7.51(d,j＝8.2hz,1h),6.87(t,j＝8.2hz,1h),6.68(d,j＝7.4hz,1h),6.46(td,j＝4.0,55.5hz,1h),5.82-5.73(m,1h),3.93-3.77(m,2h),3.73(s,3h),3.59(d,j＝8.4hz,1h),0.78(t,j＝7.2hz,3h)；¹³cnmr(100mhz,cdcl3)δ171.5,167.6,142.0,137.0,134.4,124.4,123.9,112.7(t,j＝243.8hz),103.2,98.7,94.2(t,j＝22.3hz),62.1,44.7(t,j＝2.9hz),31.0,13.6；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.07(abdd,j＝6.5,54.4,295.5hz,minor),-116.05(abdd,j＝6.9,55.8,295.3hz,minor),-121.92(abdd,j＝8.9,54.2,295.0hz,major),-125.78(abdd,j＝16.5,55.3,294.9hz,major)；hrms(esi-tof)calcd.forc15h15brf2n3o3[m+h]⁺:402.0259；found:402.0264.

本实施例制备化合物4o：黄色固体，产率58％；熔点:93.9–95.0℃；核磁共振和高分辨质谱测试分析结果如下：93:7dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ7.41(t,j＝7.6hz,1h),7.03(t,j＝7.6hz,1h),6.96(d,j＝7.6hz,1h),6.80(d,j＝7.3hz,1h),6.43(td,j＝3.6,55.1hz,1h),5.89-5.79(m,1h),3.59(d,j＝8.0hz,1h),3.35(s,3h),3.34(s,3h)；¹³cnmr(100mhz,cdcl3)δ170.8,168.6,144.5,131.5,124.7,123.4,121.5,113.0(t,j＝243.6hz),109.2,99.6,94.5(t,j＝22.2hz),52.9,44.1(t,j＝2.9hz),27.2；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.08(abdd,j＝5.7,54.0,294.2hz,minor),-115.89(abdd,j＝6.8,55.7,294.6hz,minor),-121.36(abdd,j＝9.0,54.5,295.1hz,major),-125.38(abdd,j＝16.0,55.3,295.0hz,major)；hrms(esi-tof)calcd.forc14h13f2n3nao3[m+na]⁺:332.0817；found:332.0813.

本实施例制备化合物4p：橘红色固体，产率76％；熔点:43.2–45.0℃；核磁共振和高分辨质谱测试分析结果如下：88:12dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ(major+minor)7.42-7.37(m,0.88h),7.34-7.31(m,0.12h),7.05-6.99(m,1h),6.95-6.90(m,1h),6.81-6.78(m,1h),6.60-6.30(m,1h),5.84-5.75(m,1h),4.12-3.98(m,0.24h),3.74-3.65(m,1.76h),3.62-3.59(m,0.88h),3.34(s,0.36h),3.33(s,2.64h),3.27-3.26(m,0.12h),1.24-1.01(m,2h),0.87-0.82(m,0.36h),0.56-0.52(m,2.64h)；¹³cnmr(100mhz,cdcl3)δ(major+minor)170.9,168.0,144.6,131.5,128.6,124.8,123.4,122.8(d,j＝3.2hz),121.5,112.9(t,j＝243.5hz),109.1,108.5,99.3,94.0(t,j＝22.1hz),67.5,44.3(t,j＝2.8hz),27.1,26.9,21.9,21.4,10.3,9.9；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.03(abdd,j＝5.8,54.8,294.0hz,minor),-115.88(abdd,j＝6.8,55.7,294.1hz,minor),-121.84(abdd,j＝9.4,55.0,295.0hz,major),-125.65(abdd,j＝16.3,55.4,294.6hz,major)；hrms(esi-tof)calcd.forc16h17f2n3nao3[m+na]⁺:360.1130；found:360.1141.

本实施例制备化合物4q：黄色固体，产率74％；熔点:58.1–60.1℃；核磁共振和高分辨质谱测试分析结果如下：93:7dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ7.40(t,j＝7.8hz,1h),7.01(t,j＝7.6hz,1h),6.95(d,j＝7.0hz,1h),6.78(d,j＝7.5hz,1h),6.48(td,j＝3.6,55.0hz,1h),5.65-5.55(m,1h),3.56(d,j＝9.1hz,1h),3.34(s,3h),0.98(s,9h)；¹³cnmr(100mhz,cdcl3)δ171.3,166.4,144.7,131.4,125.1,123.4,121.7,112.9(t,j＝243.4hz),108.9,98.7,92.7(t,j＝22.0hz),82.9,45.6(t,j＝2.8hz),27.4,27.0；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-110.96(abdd,j＝5.4,55.4,294.0hz,minor),-115.88(abdd,j＝6.8,56.0,294.0hz,minor),-122.87(abdd,j＝9.8,54.3,293.5hz,major),-125.97(abdd,j＝16.2,55.2,293.5hz,major)；hrms(esi-tof)calcd.forc17h19f2n3nao3[m+na]⁺:374.1287；found:374.1284.

本实施例制备化合物4r：黄色固体，产率76％；熔点:78.1–80.0℃；核磁共振和高分辨质谱测试分析结果如下：94:6dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ7.38(t,j＝7.8hz,1h),7.01-6.95(m,2h),6.78(d,j＝7.5hz,1h),6.46(td,j＝3.6,54.5hz,1h),5.81-5.71(m,1h),3.99-3.90(m,1h),3.86-3.70(m,3h),3.60(d,j＝8.5hz,1h),1.34(t,j＝7.2hz,3h),0.68(t,j＝7.2hz,3h)；¹³cnmr(100mhz,cdcl3)δ170.6,167.8,143.7,131.4,125.0,123.2,121.8,112.9(t,j＝243.5hz),109.1,99.1,93.6(t,j＝22.1hz),61.8,44.4(t,j＝2.8hz),35.7,13.5,12.6；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-111.06(abdd,j＝5.8,54.5,294.2hz,minor),-115.93(abdd,j＝7.2,55.6,294.2hz,minor),-122.03(abdd,j＝9.1,54.2,294.3hz,major),-125.82(abdd,j＝16.5,55.3,294.4hz,major)；hrms(esi-tof)calcd.forc16h17f2n3nao3[m+na]⁺:360.1130；found:360.1120.

本实施例制备化合物4s：黄色固体，产率78％；熔点:77.6–79.1℃；核磁共振和高分辨质谱测试分析结果如下：93:7dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ7.38-7.26(m,6h),7.00-6.95(m,1h),6.82-6.77(m,2h),6.52(td,j＝3.8,54.7hz,1h),5.86-5.77(m,1h),5.25(d,j＝15.7hz,1h),4.84(d,j＝15.7hz,1h),3.89-3.81(m,1h),3.77-3.69(m,2h),0.54(t,j＝7.0hz,3h)；¹³cnmr(100mhz,cdcl3)δ171.3,167.8,143.6,134.9,131.3,129.0,128.1,127.4,124.9,123.4,121.7,112.8(t,j＝243.6hz),110.1,99.2,93.8(t,j＝22.1hz),62.0,44.6,44.5(t,j＝3.0hz),13.31；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-110.91(abdd,j＝55.8,295.0hz,minor),-116.02(abdd,j＝7.9,55.7,294.4hz,minor),-122.11(abdd,j＝9.0,54.0,294.2hz,major),-125.92(abdd,j＝16.2,55.4,294.3hz,major)；hrms(esi-tof)calcd.forc21h19f2n3nao3[m+na]⁺:422.1287；found:422.1291.

本实施例制备化合物4t：绿色固体，产率67％；熔点:95.3–97.1℃；核磁共振和高分辨质谱测试分析结果如下：86:14dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ(major+minor)7.60-7.45(m,5h),7.34(t,j＝8.0hz,1h),7.06(t,j＝7.2hz,1h),6.92(d,j＝7.8hz,1h),6.86(d,j＝7.8hz,1h),6.49(t,j＝54.8hz,0.86h),6.43(td,j＝54.8,7.2hz,0.14h),5.87-5.79(m,1h),4.25-4.11(m,0.28h),3.96-3.81(m,1.72h),3.71(d,j＝8.2hz,0.86h),3.55(d,j＝8.2hz,0.14h),1.12(t,j＝6.6hz,0.42h),0.78(t,j＝6.6hz,2.58h)；¹³cnmr(100mhz,cdcl3)δ(major+minor)170.5,167.7,144.7,144.2,134.0,133.7,131.4,130.0,129.8,129.7,128.9,128.6,126.7,126.5,125.0,123.8,123.2(d,j＝11.94hz),121.4,112.8(t,j＝243.6hz),110.3,110.1,109.8,99.3,93.7(t,j＝22.2hz),62.1,62.0,45.0(t,j＝2.9hz),14.2,13.6；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-110.81(abdd,j＝6.2,54.3,294.0hz,minor),-115.92(abdd,j＝7.0,56.0,295.2hz,minor),-121.97(abdd,j＝8.8,54.5,294.8hz,major),-125.73(abdd,j＝16.2,55.2,294.4hz,major)；hrms(esi-tof)calcd.forc20h17f2n3nao3[m+na]⁺:408.1130；found:408.1129.

本实施例制备化合物4u：黄色固体，产率31％；熔点:84.6–86.5℃；核磁共振和高分辨质谱测试分析结果如下：93:7dr(determinedby¹⁹fnmranalysis)；¹hnmr(400mhz,cdcl3)δ8.01(d,j＝8.2hz,1h),7.44(t,j＝8.2hz,1h),7.12(t,j＝7.6hz,1h),6.78(d,j＝7.6hz,1h),6.49(td,j＝3.4,54.5hz,1h),5.86-5.76(m,1h),3.89-3.75(m,2h),3.65(dd,j＝1.0,8.4hz,1h),1.67(s,9h),0.72(td,j＝1.0,7.2hz,3h)；¹³cnmr(100mhz,cdcl3)δ169.4,167.5,148.7,140.9,131.7,125.1,124.5,120.7,115.9,112.6(t,j＝243.8hz),99.8,94.1(t,j＝22.2hz),85.7,62.2,45.0(t,j＝2.5hz),28.2,13.5；¹⁹fnmr(376mhz,cdcl3)δ(major+minor)-110.69(abdd,j＝5.8,54.9,296.3hz,major),-112.80(abdd,j＝8.2,56.8,299.1hz,minor),-114.77(abdd,j＝7.5,53.3,299.0hz,minor),-115.99(abdd,j＝7.4,55.5,296.0hz,major)；hrms(esi-tof)calcd.forc19h21f2n3nao5[m+na]⁺:432.1341；found:432.1337.

本发明的式(1)化合物具有重要的生物活性，体外对人前列腺(pc-3)和人肺癌细胞(a549)共两株肿瘤细胞的细胞毒性试验表明：此类式(1)所示的含二氟甲基基团的3,3'-螺环氧化吲哚对肿瘤细胞生长具有抑制作用，有可能发展成为新的防治肿瘤药物。

本发明的式(1)化合物或其可药用盐及其溶剂化物可以与药学上常用的辅料或载体结合，制备得到具有肿瘤细胞生长抑制活性从而可以用于防治肿瘤的药物组合物。上述各类药物组合物可以是固体形式，如片剂、胶囊剂、颗粒剂、粉剂；也可以是液体形式，如注射剂、悬浮剂和乳剂等。除此之外，还可以采用现代制药界所公知的控释剂或缓释剂或纳米剂。

本发明的式(1)化合物或其可药用盐及其溶剂化物可以与现已上市的抗肿瘤药物如铂类药物顺铂(ddp)、脱氧胞昔类药物吉西他滨(gemcitabine,gemzar,健择)、紫杉醇(paclitaxel)、长春花碱类药物失碳长春花碱(vinorebine,nvb诺维本)、喜树碱类药物伊立替康(irinatecan、cpt-11)、足叶乙甙(etoposide)等联合使用，制备得到具有肿瘤生长抑制活性的细胞毒性组合物，可用于治疗肿瘤疾病。该类药物组合物可以是固体形式，如片剂、胶囊剂、颗粒剂、粉剂；也可以是液体形式，如注射剂、悬浮剂和乳剂等。除此之外，还可以采用现代制药界所公知的控释剂或缓释剂或纳米剂。

药理实施例1：化合物4e、4h、4k、4q或4r对pc-3细胞的细胞毒性

pc-3(人前列腺癌)细胞用rpmi-1640培养基培养，培养基中含10％的胎牛血清，100u/ml青霉素及100u/ml的链霉素。细胞以每孔5000个细胞的浓度加入到96孔中，在37℃含5％co2潮湿空气的培养箱中培养24小时。

细胞存活率的测定用改良mtt法。细胞经过24小时的孵育后，分别将新配的化合物4e、4h、4k、4q或4r的二甲基亚砜溶液以浓度梯度加入到各孔中，使孔中化合物最终浓度分别为6.25μmol/l,12.5μmol/l,25μmol/l,50μmol/l和100μmol/l。48小时后，每孔加入10μlmtt(5mg/ml)的磷酸盐缓冲液，再继续在37℃培养4小时后，离心5分钟除去未转化的mtt，每孔中加入150μl二甲基亚砜。以溶解还原的mtt晶体甲臜(formazan)，用酶标仪在490nm波长测定od值。其中化合物4e、4h、4k、4q或4r对pc-3细胞半抑制浓度ic50由spss软件(19版本)分析得到。化合物4e对pc-3肿瘤细胞的ic50为82.7μmol/l；化合物4h对pc-3肿瘤细胞的ic50为64.6μmol/l；化合物4k对pc-3肿瘤细胞的ic50为73.9μmol/l；化合物4q对pc-3肿瘤细胞的ic50为33.7μmol/l；化合物4r对pc-3肿瘤细胞的ic50为83.2μmol/l；而阳性对照顺铂对pc-3肿瘤细胞的ic50为23.7μmol/l。

实验结论：pc-3细胞是测试化合物对肿瘤细胞的细胞毒性的有效工具和评价指标。本实验表明此类式(1)所示的含二氟甲基基团的3,3'-螺环氧化吲哚对pc-3细胞具有一定的细胞毒性，有可能发展成新的具有抗肿瘤作用的药物。

药理实施例2：化合物4f、4h、4k、4q或4t对a549细胞的细胞毒性

a549(人肺癌细胞)用dmem培养基培养，培养基中含10％的胎牛血清，100u/ml的青霉素和100u/ml链霉素。细胞以每孔4000个细胞的浓度加入到96孔中，在37℃含5％co2潮湿空气的培养箱中培养24小时。

细胞存活率的测定用改良mtt法。具体方法如药理实施例1。化合物4f对a549肿瘤细胞的ic50为55.1μmol/l；化合物4h对a549肿瘤细胞的ic50为78.2μmol/l；化合物4k对a549肿瘤细胞的ic50为88.6μmol/l；化合物4q对a549肿瘤细胞的ic50为85.7μmol/l；化合物4t对a549肿瘤细胞的ic50为89.9μmol/l；而阳性对照顺铂对a540肿瘤细胞的ic50为23.0μmol/l。

实验结论：a549细胞是测试化合物对肿瘤细胞的细胞毒性的有效工具和评价指标。本实验表明此类式(1)所示的含二氟甲基基团的3,3'-螺环氧化吲哚对a549细胞具有一定的细胞毒性，有可能发展成新的具有抗肿瘤作用的药物。

从以上药理实施例中我们可以看出该类化合物对这两株肿瘤细胞都显示有一定的细胞毒性。可见这些化合物具有开发成为抗肿瘤药物的潜力，值得继续深入研究。