本發(fā)明涉及化學(xué)技術(shù)領(lǐng)域，尤其是一種合成六氫吡啶-2,3-并吲哚-2-酮骨架化合物的關(guān)鍵中間體及其制備方法及應(yīng)用。

背景技術(shù)：

六氫吡啶-2,3-并吲哚-2-酮骨架廣泛存在于許多具有重要生物活性的吲哚生物堿中，例如neoxaline(i)1979年從一個(gè)日本曲霉培養(yǎng)液fg-551中分離出來(lái)，測(cè)試顯示具有抗癌活性，機(jī)制研究顯示；該化合物能抑制細(xì)胞增殖，在細(xì)胞分裂g期阻滯細(xì)胞進(jìn)程；至目前為止，10種相關(guān)的天然產(chǎn)物9-epi-neoxaline(ii),oxaline(iii),theglandicolins(iv,v),和themeleagrins(vi-x)也已經(jīng)在對(duì)青霉菌的培養(yǎng)液中分離出來(lái)，測(cè)試顯示也都具有抗癌活性；kapakahinea(xi)也包含六氫吡啶-2,3-并吲哚-2-酮骨架，具有抗腫瘤活性(如圖7所示)。在這個(gè)背景下，鑒于含六氫吡啶-2,3-并吲哚-2-酮天然產(chǎn)物具有潛在的生物活性，因此，合成六氫吡啶-2,3-并吲哚-2-酮骨架化合物的關(guān)鍵中間體及其制備方法，可以為生物活性篩選提供化合物源，對(duì)尋找新型的抗腫瘤活性先導(dǎo)化合物具有重要的意義(如圖8和圖9所示)。

技術(shù)實(shí)現(xiàn)要素：

本發(fā)明的目的是：提供一種合成六氫吡啶-2,3-并吲哚-2-酮骨架化合物的關(guān)鍵中間體及其制備方法，六氫吡啶-2,3-并吲哚-2-酮骨架化合物是一類(lèi)重要的抗腫瘤活性先導(dǎo)化合物，對(duì)藥物篩選和制藥行業(yè)具有重要的應(yīng)用價(jià)值，且其合成方法非常經(jīng)濟(jì)簡(jiǎn)便。

本發(fā)明是這樣實(shí)現(xiàn)的合成六氫吡啶-2,3-并吲哚-2-酮骨架化合物的關(guān)鍵中間體，該化合物具有如通式(ⅰ)所示的結(jié)構(gòu)：

式中，r¹為芐基或各種取代的芐基；r²為h或鹵素。

合成六氫吡啶-2,3-并吲哚-2-酮骨架化合物的關(guān)鍵中間體的制備方法，其特征在于：在堿性條件催化下,由相應(yīng)的3-單取代氧化吲哚1與硝基異惡唑烯烴2發(fā)生michael加成反應(yīng)得到；

合成路線如下：

式中，r¹為芐基或各種取代的芐基；r²為h或鹵素。

一種如權(quán)利要求1所述的化合物作為合成六氫吡啶-2,3-并吲哚-2-酮骨架化合物關(guān)鍵中間體；合成路線為：

式中，r¹為芐基或各種取代的芐基；r²為h或鹵素。

通過(guò)采用上述技術(shù)方案，在堿性催化條件下，以相應(yīng)的3-單取代氧化吲哚1與硝基異惡唑烯烴2發(fā)生michael加成反應(yīng)，生成產(chǎn)物3。它是合成六氫吡啶-2,3-并吲哚-2-酮骨架化合物的關(guān)鍵中間體，六氫吡啶-2,3-并吲哚-2-酮骨架化合物對(duì)藥物篩選和制藥行業(yè)具有重要的應(yīng)用價(jià)值。本發(fā)明操作簡(jiǎn)單易行，原料合成便宜易得，可以在各種有機(jī)溶劑中進(jìn)行，也具有較好的空氣穩(wěn)定性，適用性廣，對(duì)于各種取代基都有很好的兼容性。

附圖1-6為本發(fā)明的實(shí)施例化合物3aa，3ab和7cg的核磁共振譜圖；

附圖7-9為本發(fā)明的技術(shù)背景說(shuō)明圖。

具體實(shí)施方式

(一)、六氫吡啶-2,3-并吲哚-2-酮骨架化合物的關(guān)鍵中間體3的制備

化合物3aa:在反應(yīng)瓶中加入94.8mgn-甲基-3-芐基氧化吲哚1a(0.4mmol)和138mg硝基異惡唑烯烴2a(0.6mmol)，溶解在5.0ml的二氯甲烷中，加入催化劑tbab(10mol％,12.8mg)andk2co3(20mol％,11.0mg)，在氬氣保護(hù)下50℃攪拌反應(yīng)24h,tlc檢測(cè)完全后，直接經(jīng)硅膠柱層析[洗脫劑：v(乙酸乙酯):v(石油醚)＝10:1～5:1]，分離得到黃色液體3aa171.8mg,產(chǎn)率92％,2:1dr。核磁共振和高分辨質(zhì)譜測(cè)試結(jié)果如下：白色固體；m.p.151.6-152.1℃；yield92％；2:1dr；¹hnmr(cdcl3,400mhz)δ(major+minor):2.36(s,1.8h,minor),2.40(s,3h,major),2.66(s,1.8h,minor),2.85(s,3h,major),2.88-2.91(m,1h,major),3.18(d,j＝10.4hz,0.6h,minor),3.26(d,j＝10.4hz,0.6h,minor),3.48(d,j＝10.4hz,1h,major),3.51-3.52(m,1h,major),3.83-3.87(m,1h,minor),3.95-4.05(m,2h,major),6.37(d,j＝6.4hz,0.6h,minor),6.42(d,j＝6.4hz,1.0h,major),6.72-6.77(m,3.2h,major+minor),6.92-6.97(m,4.6h,major+minor),6.99-7.02(m,1h,major),7.08-7.18(m,11h,major+minor),7.42(d,j＝6.0hz,0.6h,minor)；¹³cnmr(cdcl3,100mhz)δ(major+minor):11.5,11.6,25.5,25.8,28.2,29.4,41.7,42.4,49.2,49.9,57.7,58.0,107.8,107.9,121.9,126.4,127.3,127.4,127.8,127.9,128.3,129.7,129.8,143.7,155.2,172.9,173.3,176.7,177.7；hrms(esi-tof)m/z:calcd.forc28h25n3nao4[m+na]⁺:490.1743；found:490.1745.

通過(guò)實(shí)施例制備的化合物3ab～3b’b的制備方法同化合物3aa，投料比與化合物3aa相同，可得到化合物3ab～3b’b，反應(yīng)產(chǎn)率見(jiàn)表1和表2，但需強(qiáng)調(diào)的是實(shí)施例旨在闡述而不是限制本發(fā)明的范圍。本發(fā)明的化合物不限于表1與表2所表示的內(nèi)容。

表1為合成六氫吡啶-2,3-并吲哚-2-酮骨架化合物的關(guān)鍵中間體3的化學(xué)結(jié)構(gòu)

表2為合成六氫吡啶-2,3-并吲哚-2-酮骨架化合物的關(guān)鍵中間體3的化學(xué)結(jié)構(gòu)

本實(shí)施例制備中間體化合物3ab:whitesoild,m.p.152.1-152.6℃；yield85％；20:1dr；¹hnmr(cdcl3,400mhz)δ:2.42(s,3h),2.85(s,3h),2.94(d,j＝12.4hz,1h),3.51(d,j＝12.8hz,1h),3.57-3.61(m,1h),3.98-4.10(m,2h),6.39(d,j＝7.6hz,1h),6.74-6.76(m,2h),6.80-6.84(m,2h),6.93-7.03(m,4h),7.07-7.11(m,3h),7.17(d,j＝7.2hz,1h)；¹³cnmr(cdcl3,100mhz)δ:11.6,25.8,28.1,42.1,48.6,57.7,107.9,114.9(d,jcf＝21.3hz),122.0,124.6,126.4,127.4,128.4,129.7,135.0,143.5,155.3,161.8(d,jcf＝225.8hz),173.2,177.5；hrms(esi-tof)m/z:calcd.forc28h24fn3nao4[m+na]⁺:508.1649；found:508.1651.

本實(shí)施例制備中間體化合物3ac:whitesoild,m.p.147.2-147.7℃；yield81％；8:7dr；¹hnmr(cdcl3,400mhz)δ(major+minor):2.38(s,3.0h,major),2.43(s,2.5h,minor),2.69(s,3.0h,major),2.86(s,2.6h,minor),2.92(d,j＝12.4hz,0.8h,minor),3.16(d,j＝12.8hz,1h,major),3.23(d,j＝12.8hz,1h,major),3.49(d,j＝12.8hz,0.8h,minor),3.55-3.62(m,0.8h,minor),3.78-3.81(m,2.0h,major),3.95-4.02(m,2.6h,major+minor),6.39-6.42(m,1.8h,major+minor),6.73-6.76(m,3.8h,major+minor),6.82-6.90(m,3.8h,major+minor),6.93-7.04(m,8.5h,major+minor),7.08-7.20(m,5.6h,major+minor),7.44(d,j＝7.2hz,0.9h,minor)；¹³cnmr(cdcl3,100mhz)δ(major+minor):11.5,11.6,25.5,25.8,28.2,29.5,41.8,42.3,48.9,49.4,57.5,57.8,107.9,108.0,114.7(d,jcf＝21.2hz),122.2(d,jcf＝20.2hz),123.7,124.7,126.5,127.4,128.5,128.6,129.7,129.8,134.9,155.3,160.9(d,jcf＝244.1hz),172.4,173.0,176.5,177.4；hrms(esi-tof)m/z:calcd.forc28h24fn3nao4[m+na]⁺:508.1649；found:508.1648.

本實(shí)施例制備中間體化合物3ad:whitesoild,m.p.167.6-168.1℃；yield85％；13:1dr；¹hnmr(cdcl3,400mhz)δ:2.44(s,3h),2.91(s,3h),3.14(d,j＝10.0hz,1h),3.77(d,j＝10.0hz,1h),3.95-3.98(m,1h),4.08-4.14(m,1h),4.76-4.80(m,1h),6.32(d,j＝6.0hz,1h),6.78-6.80(m,2h),6.93-7.00(m,6h),7.04-7.07(m,2h),7.29-7.31(m,1h),7.34(d,j＝5.6hz,1h)；¹³cnmr(cdcl3,100mhz)δ:11.6,25.8,28.8,42.0,43.7,57.6,107.4,121.8,124.4,126.5,126.6,127.4,128.0,128.2,128.6,129.4,129.9,143.0,155.3,173.3,177.6；hrms(esi-tof)m/z:calcd.forc28h24cln3nao4[m+na]⁺:524.1353；found:524.1357.

本實(shí)施例制備中間體化合物3ae:whitesoild,m.p.170.1-170.8℃；yield86％；9:8dr；¹hnmr(cdcl3,400mhz)δ(major+minor):2.33(s,3h,major),2.46(s,2.6h,minor),2.85(s,3h,major),2.90(s,2.6h,minor),2.95(d,j＝10.0hz,1h,major),3.03(d,j＝10.0hz,1h,major),3.13(d,j＝10.0hz,0.9h,minor),3.47(d,j＝6.4hz,2h,major),3.75(d,j＝10.0hz,0.9h,minor),3.96-4.00(m,0.9h,minor),4.08-4.10(m,0.9h,minor),4.71-7.74(m,0.9h,minor),4.81-4.84(m,1.0h,major),6.33(d,j＝6.0hz,0.9h,minor),6.43(d,j＝6.0hz,1.0h,major),6.63(d,j＝6.0hz,2.0h,major),6.78-6.80(m,1.8h,minor),6.89-7.06(m,9.0h,major+minor),7.07-7.11(m,0.9h,minor),7.14-7.17(m,1.0h,major),7.25(s,1.0h,major),7.31-7.33(m,1.8h,major+minor),7.38-7.39(m,0.9h,minor),7.43-7.44(m,1.0h,major),7.98(d,j＝6.8hz,1.0h,major)；¹³cnmr(cdcl3,100mhz)δ(major+minor):11.4,11.6,25.7,25.8,28.6,30.3,42.0,43.1,43.3,57.1,57.5,107.6,107.9,122.7,127.2,127.4,127.5,128.4,129.2,129.6,129.8,134.6,142.9,143.5,155.3,155.4,171.8,173.0,176.6,177.4；hrms(esi-tof)m/z:calcd.forc28h23cl2n3nao4[m+na]⁺:558.0963；found:558.0965.

本實(shí)施例制備中間體化合物3ag:whitesoild,m.p.164.3-164.9℃；yield91％；9:8dr；¹hnmr(cdcl3,400mhz)δ:1.26(s,9h),2.41(s,3h),2.83-2.84(m,4h),3.31-3.41(m,2h),3.78-3.84(m,1h),4.02-4.06(m,1h),6.45(d,j＝7.6hz,1h),6.68-6.70(m,2h),6.89-6.96(m,3h),7.02-7.06(m,1h),7.12-7.15(m,3h),7.18-7.23(m,3h)；¹³cnmr(cdcl3,100mhz)δ:11.6,25.8,28.4,31.2,34.4,42.6,48.5,57.9,107.8,121.9,124.8,125.3,126.3,127.3,128.3,129.0,129.7,133.7,135.3,143.9,150.5,155.2,173.3,177.9；hrms(esi-tof)m/z:calcd.forc32h33n3nao4[m+na]⁺:546.2369；found:546.2371.

本實(shí)施例制備中間體化合物3ba:whitesoild,m.p.201.4-201.7℃；yield84％；2:1dr；¹hnmr(cdcl3,400mhz)δ(major+minor):2.36(s,1.9h,minor),2.40(s,3.0h,major),2.69(s,1.9h,minor),2.86-2.88(m,4h,major),3.14(d,j＝12.8hz,0.6h,minor),3.21(d,j＝12.8hz,0.6h,minor),3.42-3.48(m,2h),3.81-3.84(m,1h,major),3.90-4.04(m,2.6h,major+minor),6.42(d,j＝8.0hz,0.6h,minor),6.46(d,j＝7.8hz,1h,major),6.65(d,j＝8.4hz,2h,major),6.67-6.71(m,1.3h,minor),6.89-6.93(m,3h,major+minor),7.00-7.04(m,1.0h,major),7.08-7.18(m,11.4h,major+minor),7.40(d,j＝7.2hz,0.6h,minor)；¹³cnmr(cdcl3,100mhz)δ(major+minor):11.5,11.6,25.5,25.8,28.2,29.3,40.9,41.6,49.2,49.9,57.6,57.8,103.0,108.1,108.2,122.0,124.9,127.5,127.6,127.7,127.8,127.9,128.0,128.5,129.1,131.0,131.1,133.7,155.2,172.7,173.1,176.4,177.5；hrms(esi-tof)m/z:calcd.forc28h24cln3nao4[m+na]⁺:524.1353；found:524.1357.

本實(shí)施例制備中間體化合物3bc:whitesoild,m.p.156.1-156.7vc；yield79％；3:2dr；¹hnmr(cdcl3,400mhz)δ(major+minor):2.38(s,2.0h,minor),2.43(s,3.0h,major),2.72(s,2.0h,minor),2.86-2.90(m,4h,major),3.12(d,j＝12.8hz,0.6h,minor),3.19(d,j＝12.8hz,0.6h,minor),3.45(d,j＝12.4hz,1h,major),3.53-3.56(m,1h,major),3.76-3.78(m,1.3h,minor),3.92-4.04(m,2.6h,major+minor),6.44-6.48(m,1.6h,major+minor),6.66-6.70(m,3.3h,major+minor),6.83-6.89(m,3.0h,major+minor),6.92(d,j＝8.4hz,5.0h,major+minor),7.02-7.06(m,1h,major),7.10-7.21(m,5.0h,major+minor),7.42(d,j＝6.4hz,0.6h,minor)；¹³cnmr(cdcl3,100mhz)δ(major+minor):11.4,11.5,25.6,25.8,28.1,29.4,41.0,41.5,49.0,49.5,57.4,57.6,108.2,108.3,114.8(d,jcf＝21.3hz),122.3(d,jcf＝19.9hz),123.6,124.6,127.6,128.7,128.8,131.1,131.2,132.4,133.5,155.3,160.9,162.1(d,jcf＝240.8hz),172.3,172.8,177.2；hrms(esi-tof)m/z:calcd.forc28h23clfn3nao4[m+na]⁺:542.1259；found:542.1259.

本實(shí)施例制備中間體化合物3bd:whitesoild,m.p.179.3-179.9℃；yield72％；3:1dr；¹hnmr(cdcl3,400mhz)δ(major+minor):2.32(s,1.2h,minor),2.44(s,3.0h,major),2.89(s,1.2h,minor),2.93(s,3.4h,major+minor),3.03(d,j＝12.8hz,0.3h,minor),3.09(d,j＝12.8hz,1.0h,major),3.50(d,j＝8.0hz,1.0h,major),3.73(d,j＝12.8hz,1.0h,major),3.89-3.94(m,1.0h,major),4.02-4.05(m,1.0h,minor),4.74-4.78(m,1.0h,major),4.84-4.88(m,0.3h,minor),6.37(d,j＝7.8hz,1.0h,major),6.48(d,j＝7.8hz,0.3h,minor),6.57(s,0.3h,minor),6.59(s,0.4h,minor),6.72(s,1.0h,major),6.74(s,1.0h,major),6.86(s,0.3h,minor),6.88(s,0.3h,minor),6.93-6.99(m,4.0h,major),7.02-7.10(m,3.4h,major+minor),7.15-7.17(m,0.3h,minor),7.19-7.15(m,0.4h,minor),7.26-7.30(m,1.4h,major+minor),7.32-7.37(m,1.8h,major+minor),7.41(d,j＝6.4hz,0.4h),7.96-7.99(m,0.4h,minor)；¹³cnmr(cdcl3,100mhz)δ(major+minor):11.4,11.6,25.8,28.8,41.1,41.2,43.5,43.7,57.0,57.5,107.7,108.0,122.0,124.3,126.6,127.6,128.0,128.4,128.7,129.5,131.2,142.9,143.4,155.3,172.0,173.1,176.4,177.4；hrms(esi-tof)m/z:calcd.forc28h23cl2n3nao4[m+na]⁺:558.0963；found:558.0960.

本實(shí)施例制備中間體化合物3be:whitesoild,m.p.188.5-188.9℃；yield76％；5:1dr；¹hnmr(cdcl3,400mhz)δ(major+minor):2.33(s,0.8h,minor),2.46(s,3.0h,major),2.89(s,0.8h,minor),2.93(s,3.0h,major),2.99(d,j＝0.2h,minor),3.08(d,j＝12.8hz,1.0h,major),3.45(d,j＝8.0hz,0.5h,minor),3.71(d,j＝12.4hz,1.3h,major+minor),3.91-3.96(m,1.0h,major),4.04(d,j＝11.6hz,1.0h,major),4.69-4.73(m,1.0h,major),4.79-4.83(m,0.3h,minor),6.38(d,j＝7.6hz,1h,major),6.48(d,j＝8.0hz,0.2h,minor),6.57(d,j＝8.4hz,0.5h,minor),6.71(s,1h),6.73(s,1h),6.88(d,j＝8.4hz,0.5h,minor),6.93-6.98(m,3.0h,major),7.04-7.07(m,3.0h,major),7.11(d,j＝7.6hz,0.2h,minor),7.18(d,j＝7.2hz,0.2h,minor),7.22-7.25(m,1.0h,major),7.30-7.33(m,1.3h,major+minor),7.38-7.39(m,0.2h,minor),7.43(d,j＝7.2hz,0.3h,major+minor),7.95(d,j＝8.4hz,0.2h,minor)；¹³cnmr(cdcl3,100mhz)δ(major+minor):11.6,25.9,28.6,41.1,43.3,57.4,107.9,108.2,122.1,124.2,127.0,127.6,128.7,128.9,129.3,131.1,133.3,142.8,155.4,172.8,177.2；hrms(esi-tof)m/z:calcd.forc28h22cl3n3nao4[m+na]⁺:592.0574；found:592.0575.

本實(shí)施例制備中間體化合物3bh:whitesoild,m.p.186.7-187.2℃；yield79％；>20:1dr；¹hnmr(cdcl3,400mhz)δ:2.38(s,3h),2.73(s,3h),3.08-3.19(m,2h),3.69-3.79(m,2h),3.89-3.93(m,1h),6.45(d,j＝7.6hz,1h),6.68(d,j＝8.0hz,2h),6.92(d,j＝8.4hz,2h),7.03-7.32(m,6h),7.41(d,j＝7.2hz,1h)；¹³cnmr(cdcl3,100mhz)δ:11.5,25.6,29.3,41.0,49.4,57.6,108.3,121.8,122.4,123.5,127.6,128.4,128.8,129.5,130.9,131.1,132.4,133.4,139.5,143.7,155.3,172.2,176.2；hrms(esi-tof)m/z:calcd.forc28h23brcln3nao4[m+na]⁺:602.0458；found:602.0459.

本實(shí)施例制備中間體化合物3bj:whitesoild,m.p.199.2-199.9℃；yield85％；20:1dr；¹hnmr(cdcl3,400mhz)δ:2.18(s,3h),2.23(s,3h),2.35(s,3h),2.79(s,3h),3.08(d,j＝12.8hz,1h),3.26(d,j＝12.4hz,1h),3.69-3.75(m,1h),3.82-3.88(m,1h),4.23-4.27(m,1h),6.47(d,j＝7.6hz,1h),6.66(d,j＝8.4hz,2h),6.86-6.91(m,4h),6.99-7.04(m,1h),7.10(d,j＝7.2hz,1h),7.15-7.20(m,2h)；¹³cnmr(cdcl3,100mhz)δ:11.5,20.1,20.9,25.7,29.7,40.2,42.6,57.4,108.1,122.2,123.8,126.7,127.5,127.8,128.5,131.2,131.3,133.3,133.7,143.7,155.3,173.1,176.9；hrms(esi-tof)m/z:calcd.forc30h28cln3nao4[m+na]⁺:552.1666；found:552.1667.

本實(shí)施例制備中間體化合物3cc:whitesoild,m.p.149.9-150.3℃；yield84％；3:2dr；¹hnmr(cdcl3,400mhz)δ(major+minor):2.38(s,2.2h,minor),2.43(s,3.0h,major),2.72(s,2.2h,minor),2.89(s,3.6h,major+minor),3.10(d,j＝12.8hz,0.7h,minor),3.18(d,j＝12.4hz,0.7h,minor),3.45(d,j＝12.4hz,1.0h,major),3.52-3.56(m,1.0h,major),3.75-3.78(m,1.6h,major+minor),3.92-4.01(m,2.8h,major+minor),6.44-6.48(m,1.7h,major+minor),6.60-6.64(m,3.6h,major+minor),6.83-6.93(m,5.4h,major+minor),7.02-7.23(m,9.9h,major+minor),7.42(d,j＝7.2hz,0.8h)；¹³cnmr(cdcl3,100mhz)δ(major+minor):11.5,11.6,25.6,25.9,28.1,29.7,41.1,41.5,49.0,49.5,57.3,57.6,108.2,108.3,114.8(d,jcf＝21.4hz),120.6,122.3(d,jcf＝20.1hz),123.6,124.6,128.7,128.9,129.5,130.6,131.4,131.5,155.3,162.2(d,jcf＝233.1hz),172.3,172.8,176.2,177.2；hrms(esi-tof)m/z:calcd.forc28h23brfn3nao4[m+na]⁺:586.0754；found:586.0755.

本實(shí)施例制備中間體化合物3cd:whitesoild,m.p.159.4-159.8℃；yield74％；20:1dr；¹hnmr(cdcl3,400mhz)δ:2.37(s,3h),2.86(s,3h),3.01(d,j＝12.8hz,1h),3.65(d,j＝12.8hz,1h),3.81-3.86(m,1h),3.94-4.01(m,1h),4.67-4.71(m,1h),6.30(d,j＝7.6hz,1h),6.60(d,j＝8.4hz,2h),6.86-6.92(m,2h),6.97-7.03(m,5h),7.19-7.22(m,1h),7.26(d,j＝7.2hz,1h)；¹³cnmr(cdcl3,100mhz)δ:11.6,25.8,28.8,41.2,43.7,57.4,107.7,122.0,124.3,126.6,128.0,128.5,128.7,129.5,130.5,131.6,134.0,142.9,155.3,173.1,177.3；hrms(esi-tof)m/z:calcd.forc28h23brcln3nao4[m+na]⁺:602.0458；found:602.0461.

本實(shí)施例制備中間體化合物3cg:whitesoild,m.p.161.7-162.2℃；yield90％；>20:1dr；¹hnmr(cdcl3,400mhz)δ:1.24(s,9h),2.35(s,3h),2.71(s,3h),3.06(d,j＝13.2hz,1h),3.16(d,j＝12.8hz,1h),3.75-3.77(m,2h),3.89-3.93(m,1h),6.41(d,j＝7.6hz,1h),6.62(d,j＝8.4hz,2h),7.04-7.08(m,5h),7.15-7.19(m,3h),7.36(d,j＝7.2hz,1h)；¹³cnmr(cdcl3,100mhz)δ:11.5,25.5,29.5,31.2,34.4,41.1,49.5,57.7,108.1,120.4,122.2,123.6,124.7,128.5,128.8,130.5,131.5,134.0,134.4,143.8,150.5,155.2,172.9,176.5；hrms(esi-tof)m/z:calcd.forc32h32brn3nao4[m+na]⁺:624.1474；found:624.1477.

本實(shí)施例制備中間體化合物3ch:whitesoild,m.p.174.1-174.5℃；yield89％；>20:1dr；¹hnmr(cdcl3,400mhz)δ:2.38(s,3h),2.73(s,3h),3.08(d,j＝12.8hz,1h),3.15(d,j＝12.8hz,1h),3.73-3.76(m,2h),3.89-3.92(m,1h),6.46(d,j＝7.6hz,1h),6.63(d,j＝8.4hz,2h),7.03-7.09(m,3h),7.11-7.15(m,2h),7.19-7.23(m,1h),7.25(s,1h),7.29-7.32(m,1h),7.40(d,j＝7.6hz,1h)；¹³cnmr(cdcl3,100mhz)δ:11.5,25.6,29.3,41.0,49.5,57.6,108.3,120.6,121.8,122.4,123.5,128.3,128.9,129.5,130.5,130.9,131.5,134.0,139.4,143.7,155.3,172.2,176.2；hrms(esi-tof)m/z:calcd.forc28h23br2n3nao4[m+na]⁺:645.9953；found:645.9957.

本實(shí)施例制備中間體化合物3ci:whitesoild,m.p.157.4-157.9℃；yield75％；20:1dr；¹hnmr(cdcl3,400mhz)δ:2.18(s,3h),2.33(s,3h),2.69(s,3h),3.06(d,j＝12.8hz,1h),3.16(d,j＝12.8hz,1h),3.75-3.77(m,2h),3.84-3.86(m,1h),6.40(d,j＝7.6hz,1h),6.61(d,j＝8.0hz,2h),6.88-6.95(m,3h),6.99-7.07(m,4h),7.13-7.17(m,1h),7.35(d,j＝6.8hz,1h)；¹³cnmr(cdcl3,100mhz)δ:11.6,21.4,25.7,29.5,41.0,49.9,57.8,108.2,120.6,122.3,123.8,127.8,128.5,128.7,130.6,131.6,134.5,137.5,144.0,155.4,172.9,176.6；hrms(esi-tof)m/z:calcd.forc29h26brn3nao4[m+na]⁺:582.1004；found:582.1001.

本實(shí)施例制備中間體化合物3cj:whitesoild,m.p.137.1-137.6℃；yield88％；>20:1dr；¹hnmr(cdcl3,400mhz)δ:2.10(s,3h),2.15(s,3h),2.27(s,3h),3.00(d,j＝12.8hz,1h),3.18(d,j＝12.8hz,1h),3.71-3.84(m,2h),4.15-4.19(m,1h),6.40(d,j＝7.2hz,1h),6.53(d,j＝8.4hz,2h),6.78(d,j＝7.2hz,2h),6.92-7.04(m,4h),7.08-7.11(m,2h)；¹³cnmr(cdcl3,100mhz)δ:11.5,20.0,20.9,25.6,30.0,40.1,42.6,57.3,108.1,122.2,123.8,126.6,127.7,128.4,130.4,131.2,131.5,133.2,134.2,143.7,155.2,173.1,176.8；hrms(esi-tof)m/z:calcd.forc30h28brn3nao4[m+na]⁺:596.1161；found:596.1165.

本實(shí)施例制備中間體化合物3da:whitesoild,m.p.132.1-132.6℃；yield90％；18:1dr；¹hnmr(cdcl3,400mhz)δ:2.34(s,3h),2.66(s,3h),3.10-3.14(m,1h),3.19(d,j＝12.8hz,1h),3.77-3.83(m,2h),3.91-3.95(m,1h),6.39(d,j＝7.6hz,1h),6.59-6.64(m,2h),6.68-6.72(m,2h),7.06-7.18(m,7h),7.39-7.41(m,1h)；¹³cnmr(cdcl3,100mhz)δ:11.5,11.6,25.5,25.8,28.2,29.4,40.8,41.6,49.1,49.8,58.0,58.5,108.0,108.1,114.1,114.2(d,jcf＝17.7hz),122.2(d,jcf＝19.6hz),123.7,127.7,127.9,128.0,131.2,131.3,143.7,143.9,155.3,161.4(d,jcf＝216.7hz),172.8,173.2,176.6；hrms(esi-tof)m/z:calcd.forc28h24fn3nao4[m+na]⁺:508.1649；found:508.1647.

本實(shí)施例制備中間體化合物3dc:whitesoild,m.p.134.2-134.7℃；yield83％；5:1dr；¹hnmr(cdcl3,400mhz)δ(major+minor):2.38(s,3.0h,major),2.43(s,0.8h,minor),2.71(s,3.0h,major),2.88(s,0.8h,minor),3.12(d,j＝10.4hz,1.0h,major),3.20(d,j＝10.4hz,1.0h,major),3.47(d,j＝10.0hz,0.3h,minor),3.53-3.57(m,0.3h,minor),3.70-3.74(m,1.0h,major),3.77-3.79(m,1.7h,major+minor),3.94-3.99(m,1.4h,major+minor),6.42-6.46(m,1.3h,major+minor),6.62-6.65(m,2.6h,major+minor),6.68-6.73(m,2.6h,major+minor),6.83-6.91(m,2.3h,major+minor),6.92-6.94(m,1.6h,major+minor),7.02-7.06(m,0.3h,minor),7.10-7.16(m,2.6h,major+minor),7.18-7.21(m,1.3h,major+minor),7.43(d,j＝5.6hz,1.0h,major)；¹³cnmr(cdcl3,100mhz)δ(major+minor):11.5,11.6,25.5,25.8,28.2,29.4,41.0,41.4,48.8,49.4,57.8,58.4,108.2,114.3(d,jcf＝17.9hz),114.8(d,jcf＝17.6hz),122.4,123.6,125.1,128.6,128.8,129.3,129.4,130.6,131.2,131.3,139.7,139.8,143.6,143.8,155.3,161.5(d,jcf＝194.3hz),162.1(d,jcf＝196.2hz),172.3,172.9,175.1,176.4；hrms(esi-tof)m/z:calcd.forc28h23f2n3nao4[m+na]⁺:526.1554；found:526.1554.

本實(shí)施例制備中間體化合物3dd:whitesoild,m.p.172.4-173.1℃；yield80％；>20:1dr；¹hnmr(cdcl3,400mhz)δ:2.32(s,3h),2.88(s,3h),2.93(d,j＝12.8hz,1h),3.03(d,j＝12.8hz,1h),3.51(d,j＝8.0hz,2h),4.85-4.89(m,1h),6.46(d,j＝7.6hz,1h),6.58-6.60(m,4h),7.07-7.11(m,1h),7.15-7.27(m,2h),7.32-7.38(m,2h),7.42(d,j＝6.8hz,1h),7.98-8.01(m,1h)；¹³cnmr(cdcl3,100mhz)δ:11.4,25.7,30.3,41.0,43.4,57.2,108.0,114.0(d,jcf＝16.9hz),122.7,123.3,127.2,128.5,129.0,129.5,131.1,131.2,143.4,155.2,161.5(d,jcf＝195.8hz),172.1,176.6；hrms(esi-tof)m/z:calcd.forc28h23clfn3nao4[m+na]⁺:542.1259；found:542.1257.

本實(shí)施例制備中間體化合物3de:whitesoild,m.p.164.1-164.8℃；yield89％；15:1dr；¹hnmr(cdcl3,400mhz)δ:2.33(s,3h),2.88(s,3h),2.90-2.93(m,1h),2.99(d,j＝12.8hz,1h),3.46(d,j＝8.0hz,2h),4.79-4.83(m,1h),6.47(d,j＝7.6hz,1h),6.59(d,j＝7.6hz,4h),7.07-7.12(m,1h),7.16-7.20(m,1h),7.27-7.34(m,1h),7.39-7.44(m,2h),7.96(d,j＝8.4hz,1h)；¹³cnmr(cdcl3,100mhz)δ:11.4,25.7,30.2,41.1,43.0,57.1,108.1,114.0(d,jcf＝21.3hz),122.8,123.2,127.5,128.6,129.2,131.1,131.2,132.0,143.4,155.3,161.5(d,jcf＝250.1hz),171.7,176.4；hrms(esi-tof)m/z:calcd.forc28h22cl2fn3nao4[m+na]⁺:576.0869；found:576.0871.

本實(shí)施例制備中間體化合物3dg:whitesoild,m.p.142.3-142.9℃；yield90％；>20:1dr；¹hnmr(cdcl3,400mhz)δ:1.27(s,9h),2.41(s,3h),2.87(s,3h),3.28-3.32(m,1h),3.35(d,j＝10.4hz,1h),3.70-3.80(m,2h),4.02-4.05(m,1h),6.50(d,j＝6.0hz,1h),6.59-6.66(m,4h),7.04-7.07(m,1h),7.13-7.20(m,4h),7.23(d,j＝6.4hz,2h)；¹³cnmr(cdcl3,100mhz)δ:11.6,25.8,28.4,31.2,34.4,41.8,48.4,57.9,108.0,114.1(d,jcf＝17.4hz),122.0,124.9,125.2,128.0,128.5,131.1,131.2,133.7,143.9,150.7,155.2,161.5(d,jcf＝195.4hz),173.2,177.8；hrms(esi-tof)m/z:calcd.forc32h32fn3nao4[m+na]⁺:564.2275；found:564.2278.

本實(shí)施例制備中間體化合物3di:whitesoild,m.p.161.7-161.9℃；yield81％；5:1dr；¹hnmr(cdcl3,500mhz)δ(major+minor):2.14(s,0.8h,minor),2.18(s,3.0h,major),2.28(s,0.8h,minor),2.34(s,3.0h,major),2.63(s,0.8h,minor),2.80(s,3.0h,major),3.02-3.06(m,0.5h,minor),3.11-3.14(m,0.2h,minor),3.11-3.34(m,2.0h,major),3.71-3.73(m,0.5h,minor),3.77-3.82(m,1.0h,major),3.90-3.93(m,1.0h,major),4.22-4.25(m,1.0h,major),6.34(d,j＝6.4hz,0.2h,minor),6.41(d,j＝6.0hz,1.0h,major),6.53-6.65(m,5.0h,major+minor),6.83-7.02(m,6.6h,major+minor),7.06-7.10(m,2.3h,major+minor),7.31(d,0.2h,j＝6.0h,minor),7.45-7.46(m,0.5h,minor),7.63-7.65(m,0.5h,minor)；¹³cnmr(cdcl3,125mhz)δ(major+minor):11.6,13.7,19.2,21.3,25.8,28.3,29.7,30.5,41.7,49.0,49.7,57.8,65.6,108.0,114.1(d,jcf＝22.3hz),122.0,125.1,127.8,128.1,128.5,128.8,130.9,131.2,136.7,137.6,143.8,155.2,161.5(d,jcf＝248.3hz),173.2,177.7；hrms(esi-tof)m/z:calcd.forc29h26fn3nao4[m+na]⁺:522.1805；found:522.1807.

本實(shí)施例制備中間體化合物3ed:whitesoild,m.p.158.7-159.3℃；yield82％；3:2dr；¹hnmr(cdcl3,400mhz)δ(major+minor):2.28(s,2.2h,minor),2.38(s,3.0h,major),2.82(s,1.9h,minor),2.83(s,3.0h,major),2.87(d,j＝10.4hz,0.8h,minor),2.95(d,j＝10.4hz,0.8h,minor),3.02(d,j＝10.0hz,1.0h,major),3.43-3.50(m,1.0h,major),3.67(d,j＝10.4hz,1.0h,major),3.85-3.89(m,1.0h,major),3.99-4.04(m,1.0h,major),4.22-4.25(m,0.6h,minor),4.63-4.66(m,0.9h,minor),4.70-4.73(m,0.8h,minor),6.15(d,j＝6.8hz,1.0h,major),6.30(d,j＝6.8hz,0.8h,minor),6.47(d,j＝6.8hz,1.6h,major+minor),6.63(d,j＝6.8hz,1.9h,major+minor),6.90-6.93(m,1.0h,major),6.99-7.02(m,3.4h,major+minor),7.06-7.08(m,2.8h,major+minor),7.16-7.19(m,2.0h,major),7.22-7.24(m,0.8h,minor),7.27-7.33(m,2.4h,major+minor),7.43(d,j＝12.0hz,1.0h,major),7.84(d,j＝5.6hz,0.7h,minor)；¹³cnmr(cdcl3,100mhz)δ(major+minor):11.4,11.6,25.9,28.6,29.7,40.8,41.0,43.4,43.7,57.2,57.7,65.6,109.1,109.5,126.4,126.7,127.5,127.8,129.0,129.2,129.6,129.7,130.6,130.8,130.9,131.3,131.4,131.5,133.6,141.9,142.5,155.3,155.4,171.7,172.9,175.9,176.8；hrms(esi-tof)m/z:calcd.forc28h22br2cln3nao4[m+na]⁺:679.9563；found:679.9565.

(二)、六氫吡啶-2,3-并吲哚-2-酮類(lèi)化合物7cg的合成制備

中間體化合物4cg:在反應(yīng)瓶中加入化合物3cg(601mg,1.0mmol)和sncl2·2h2o(677mg,3.0mmol)，溶解在20.0ml四氫呋喃中，加入20毫升水和36％hcl(1.0ml)，加熱回流15小時(shí),冷卻至室溫，減壓除去溶劑，加入水，乙酸乙酯萃取，硫酸鈉干燥，除去溶劑后直接經(jīng)硅膠柱層析[洗脫劑：v(乙酸乙酯):v(石油醚)＝5:1～2:1]，分離得到淺黃色固體4cg(81％產(chǎn)率)。

中間體化合物5cg:在反應(yīng)瓶中加入4cg(519mg,1.0mmol)，溶解在meoh(30ml)中，在冰浴下加入socl2(350μl)后，在室溫下攪拌反應(yīng)5h,旋干溶劑，直接經(jīng)硅膠柱層析[洗脫劑：v(乙酸乙酯):v(石油醚)＝5:1]，分離得到黃色液體5cg(88％產(chǎn)率)。

中間體化合物6cg:在反應(yīng)瓶中加入中間體化合物5cg(0.40mmol)，溶解在3.0ml的meoh中，加入menh2(3ml,30％inmeoh)，在室溫下攪拌反應(yīng)48h后，tlc檢測(cè)完全后，減壓蒸干溶劑，溶解在二氯甲烷中經(jīng)硅膠柱層析[洗脫劑：v(乙酸乙酯):v(石油醚)＝5:1]，分離得到黃色液體6cg(84％產(chǎn)率)。

終產(chǎn)物7cg:在反應(yīng)瓶中加入中間體化合物6cg(0.4mmol)，溶解在na干燥過(guò)的6.0ml的thf中，在0℃下加入lialh4(6.0eq,2.4mmol,91.2mg)，在0℃下攪拌反應(yīng)5h后，tlc檢測(cè)完全后，用飽和的氯化銨萃滅反應(yīng)，乙酸乙酯萃取多次后，硫酸鈉干燥后，減壓蒸干溶劑，二氯甲烷溶解經(jīng)硅膠柱層析[洗脫劑：v(乙酸乙酯):v(石油醚)＝3:1]，分離得到淡黃色固體7cg(71％產(chǎn)率)。

本實(shí)施例制備終產(chǎn)物5cg:lightyellowsoild；m.p.158.7-159.4℃；yield88％；>20:1dr；¹hnmr(cdcl3,400mhz)δ:1.15(s,9h),2.54(s,3h),2.79-2.92(m,2h),2.99(d,j＝13.2hz,1h),3.09(d,j＝12.8hz,1h),3.41(s,3h),3.64-3.71(m,1h),6.33(d,j＝8.0hz,1h),6.58(d,j＝8.4hz,2h),6.81(d,j＝8.0hz,2h),6.98-7.05(m,5h),7.09-7.13(m,1h),7.28(d,j＝7.2hz,1h)；¹³cnmr(cdcl3,100mhz)δ:25.4,31.2,34.3,35.6,40.3,48.2,51.7,58.0,107.9,120.3,121.9,124.0,124.3,128.4,128.6,130.5,131.6,134.9,144.1,149.9,172.2,177.0；hrms(esi-tof)m/z:calcd.forc30h32brnnao3[m+na]⁺:556.1463；found:556.1467.

本實(shí)施例制備終產(chǎn)物7cg:lightyellowsoild；m.p.86.7-87.1℃；yield71％；>20:1dr；；¹hnmr(cdcl3,400mhz)δ:1.26(s,9h),2.44(s,3h),2.69-2.75(m,3h),2.78-2.82(m,1h),3.04(s,3h),3.53-3.56(m,1h),4.28(s,1h),6.25(d,j＝8.0hz,1h),6.54-6.55(m,2h),6.65-6.72(m,2h),6.90(d,j＝8.4hz,2h),6.99-7.00(m,3h),7.04-7.08(m,1h),7.25(d,j＝8.4hz,2h)；¹³cnmr(cdcl3,100mhz)δ:31.3,34.4,34.5,35.3,35.7,39.9,44.4,52.1,86.5,107.6,118.0,123.6,125.2,126.4,127.7,128.6,128.9,130.0,131.4,136.4,136.9,149.7,150.0,170.7；hrms(esi-tof)m/z:calcd.forc30h34n2nao[m+na]⁺:461.2569；found:461.2573.

本發(fā)明由式(1)所述的其中化合物3cg，繼續(xù)進(jìn)一步合成的六氫吡啶-2,3-并吲哚-2-酮類(lèi)化合物7cg具有重要的生物活性，體外對(duì)人肺癌細(xì)胞(a549),人白血病細(xì)胞(k562),以及體外對(duì)人前列腺(pc-3)共三株腫瘤細(xì)胞的細(xì)胞毒性試驗(yàn)表明：六氫吡啶-2,3-并吲哚-2-酮類(lèi)化合物7cg對(duì)腫瘤細(xì)胞生長(zhǎng)具有抑制作用，有可能發(fā)展成為新的防治腫瘤藥物或防治腫瘤藥物中間體。必須說(shuō)明，本發(fā)明的藥理實(shí)施例是用于說(shuō)明本發(fā)明而不是對(duì)本發(fā)明的限制。根據(jù)本發(fā)明的實(shí)質(zhì)對(duì)本發(fā)明進(jìn)行的簡(jiǎn)單改進(jìn)都屬于本發(fā)明要求保護(hù)否認(rèn)范圍。

藥理實(shí)施例1：化合物7cg對(duì)a549細(xì)胞的細(xì)胞毒性

a549(人非小細(xì)胞肺癌肺癌)用dmem培養(yǎng)基培養(yǎng)，培養(yǎng)基中含10％的胎牛血清，100u/ml的青霉素和100u/ml鏈霉素。細(xì)胞以每孔4000個(gè)細(xì)胞的濃度加入到96孔中，在37℃含5％co2潮濕空氣的培養(yǎng)箱中培養(yǎng)24小時(shí)。

細(xì)胞存活率的測(cè)定用改良mtt法。細(xì)胞經(jīng)過(guò)24小時(shí)的孵育后，分別將新配的化合物7cg的二甲基亞砜溶液以濃度梯度加入到各孔中，使孔中化合物最終濃度分別為6.25μmol/l,12.5μmol/l,25μmol/l,50μmol/l和100μmol/l。48小時(shí)后，每孔加入10μlmtt(5mg/ml)的磷酸鹽緩沖液，再繼續(xù)在37℃培養(yǎng)4小時(shí)后，離心5分鐘除去未轉(zhuǎn)化的mtt，每孔中加入150μl二甲基亞砜。以溶解還原的mtt晶體甲臜(formazan)，用酶標(biāo)儀在490nm波長(zhǎng)測(cè)定od值。其中化合物7cg對(duì)a549細(xì)胞半抑制濃度ic50由spss軟件(19版本)分析得到?；衔?cg對(duì)a549腫瘤細(xì)胞的ic50為27.4μmol/l；而陽(yáng)性對(duì)照順鉑對(duì)a549腫瘤細(xì)胞的ic50為27.5μmol/l。

實(shí)驗(yàn)結(jié)論：a549細(xì)胞是測(cè)試化合物對(duì)腫瘤細(xì)胞的細(xì)胞毒性的有效工具和評(píng)價(jià)指標(biāo)。本實(shí)驗(yàn)表明此類(lèi)式(1)所示的六氫吡啶-2,3-并吲哚-2-酮類(lèi)化合物7cg對(duì)a549細(xì)胞具有較強(qiáng)的細(xì)胞毒性，和腫瘤治療一線用藥順鉑同一數(shù)量級(jí)，有可能發(fā)展成新的具有抗腫瘤作用的藥物。

藥理實(shí)施例2：化合物7cg對(duì)k562細(xì)胞的細(xì)胞毒性

k562(人慢性髓系白血病細(xì)胞)用rpmi-1640培養(yǎng)基培養(yǎng)，培養(yǎng)基中含10％的胎牛血清，100u/ml的青霉素和100u/ml鏈霉素。細(xì)胞以每孔5000個(gè)細(xì)胞的濃度加入到96孔中，在37℃含5％co2潮濕空氣的培養(yǎng)箱中培養(yǎng)24小時(shí)。

細(xì)胞存活率的測(cè)定用改良mtt法。具體方法如藥理實(shí)施例1?；衔?cg對(duì)k562腫瘤細(xì)胞的ic50為31.0μmol/l；而陽(yáng)性對(duì)照順鉑對(duì)k562腫瘤細(xì)胞的ic50為24.7μmol/l。

實(shí)驗(yàn)結(jié)論：k562細(xì)胞是測(cè)試化合物對(duì)腫瘤細(xì)胞的細(xì)胞毒性的有效工具和評(píng)價(jià)指標(biāo)。本實(shí)驗(yàn)表明此類(lèi)式(1)所示的六氫吡啶-2,3-并吲哚-2-酮類(lèi)化合物7cg對(duì)k562細(xì)胞具有較強(qiáng)的細(xì)胞毒性，和腫瘤治療一線用藥順鉑同一數(shù)量級(jí)，有可能發(fā)展成新的具有抗腫瘤作用的藥物或中間體。

藥理實(shí)施例3：化合物7cg對(duì)pc-3細(xì)胞的細(xì)胞毒性

pc-3(人前列腺癌)細(xì)胞用rpmi-1640培養(yǎng)基培養(yǎng)，培養(yǎng)基中含10％的胎牛血清，100u/ml青霉素及100u/ml的鏈霉素。細(xì)胞以每孔5000個(gè)細(xì)胞的濃度加入到96孔中，在37℃含5％co2潮濕空氣的培養(yǎng)箱中培養(yǎng)24小時(shí)。

細(xì)胞存活率的測(cè)定用改良mtt法。具體方法如藥理實(shí)施例1?；衔?cg對(duì)pc-3腫瘤細(xì)胞的ic50為27.1μmol/l而陽(yáng)性對(duì)照順鉑對(duì)pc-3腫瘤細(xì)胞的ic50為25.2μmol/l。

實(shí)驗(yàn)結(jié)論：pc-3細(xì)胞是測(cè)試化合物對(duì)腫瘤細(xì)胞的細(xì)胞毒性的有效工具和評(píng)價(jià)指標(biāo)。本實(shí)驗(yàn)表明此類(lèi)式(1)所示的六氫吡啶-2,3-并吲哚-2-酮類(lèi)化合物7cg對(duì)pc-3細(xì)胞具有較強(qiáng)的細(xì)胞毒性，和腫瘤治療一線用藥順鉑同一數(shù)量級(jí)，有可能發(fā)展成新的具有抗腫瘤作用的藥物。

從以上藥理實(shí)施例中我們可以看出這些六氫吡啶-2,3-并吲哚-2-酮類(lèi)化合物7cg對(duì)這三株腫瘤細(xì)胞都顯示有一定的細(xì)胞毒性?？梢?jiàn)六氫吡啶-2,3-并吲哚-2-酮類(lèi)化合物具有開(kāi)發(fā)成為抗腫瘤藥物或中間體的潛力，值得繼續(xù)深入研究下去。