CN113801073B - 14-氯-β-榄香烯一氧化氮供体型衍生物及其制备和应用 - Google Patents
14-氯-β-榄香烯一氧化氮供体型衍生物及其制备和应用 Download PDFInfo
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- CN113801073B CN113801073B CN202111180424.7A CN202111180424A CN113801073B CN 113801073 B CN113801073 B CN 113801073B CN 202111180424 A CN202111180424 A CN 202111180424A CN 113801073 B CN113801073 B CN 113801073B
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Abstract
本发明公开了14‑氯‑β‑榄香烯一氧化氮供体型衍生物及其制备和在制备抗肿瘤药物中的应用。14‑氯‑β‑榄香烯一氧化氮供体型衍生物具有如下通式(I)：
式(I)中：R1为含氮和氧原子的直链或环状醇胺结构；R2、R3分别独立选自C1‑10链烷基、C3‑12环烷基、C6‑12芳基、5‑10元环杂芳基、C2‑10烯基、C2‑10炔基或C2‑10烷氧基。本发明衍生物在设计策略上优于以往的β‑榄香烯一氧化氮供体型衍生物，引入了可提高体内抗肿瘤活性的醇胺结构作为连接臂，提高了体内稳定性，并且对临床上缺乏有效治疗药物的恶性脑胶质瘤具有很好的治疗活性。
Description
技术领域
本发明涉及药物化学领域，具体涉及一类14-氯-β-榄香烯一氧化氮供体型衍生物及其制备和应用。
背景技术
榄香烯是从莪术中提取分离的倍半萜天然产物，具有广泛抗肿瘤活性。1994年，榄香烯口服乳和注射液经国家食品药品监督管理总局(China Food and DrugAdministration，CFDA)批准，作为广谱抗癌药物用于肺癌、肝癌、食管癌、鼻咽癌和脑癌的治疗。
在榄香烯提取混合物中，β-榄香烯的含量最高，也是最主要的抗肿瘤活性成分。但是，β-榄香烯结构中只含有碳氢两种元素，导致其脂溶性过强、水溶性差、生物利用度较低，不易被人体吸收，使其临床应用受到限制。以β-榄香烯注射液为例，患者治疗过程中往往需要大剂量注射给药，药液对患者血管的刺激性较大，容易引发静脉炎。因此，有必要对β-榄香烯进行结构改造与修饰，一方面改善其理化性质，另一方面提高其抗肿瘤活性。
一氧化氮(NO)参与多种生理和病理过程，且高水平的一氧化氮可通过多种信号通路抑制肿瘤细胞的生长，如ERKs，Akt等。但是，一氧化氮作为一种气体小分子，不易定量和运输，因此制备一种便于携带、稳定性好的一氧化氮供体成为了研究的热点。
研究发现，苯磺酰基呋咱是一种经典的一氧化氮供体，可以在体内外产生高水平的一氧化氮。因此，在β-榄香烯结构中引入一氧化氮供体，将大幅提高β-榄香烯的抗肿瘤活性，改善β-榄香烯的类药性，有希望获得疗效更好的抗肿瘤药物。
中国专利申请201710066664.1公开了一种一氧化氮供体型β-榄香烯衍生物的合成，具体通式如图1所示。该专利中报道的化合物是以β-榄香烯的13-醇为中间体，借助于酯化反应与呋咱类一氧化氮供体相连接，制备获得了酯类一氧化氮β-榄香烯衍生物。该系列化合物虽然表现出很好的体外抗肿瘤活性和体内抑瘤活性，但如图1所示，该系列化合物中β-榄香烯13位的酯键在体内极易被酯酶水解，代谢生成13-β-榄香醇。随后，13-β-榄香烯醇将迅速被氧化代谢为13-β-榄香醛。而13-β-榄香醛具有很强的细胞毒性，中、高剂量给药长期可直接造成给药动物死亡，因此长期用药具有较大的安全性风险，不适于应用于人体。此外，由于该专利中公开的化合物易于代谢分解，化合物稳定性差，很难透过血脑屏障，无法有效治疗脑内肿瘤，如恶性脑胶质瘤。
发明内容
本发明所公开的化合物，以醇胺结构为连接臂，首先制备得到13-β-榄香烯胺类中间体，再与呋咱一氧化氮供体连接，最终制备得到具有新型连接臂的β-榄香烯一氧化氮供体型衍生物，具有优异活性的同时不存在专利201710066664.1中可能出现的毒性问题。
13,14-双氯-β-榄香烯的合成路线如图2所示，反应条件和试剂：N-氯代丁二酰亚胺NCS，三氟甲烷磺酸镱Yb(OTf)3，三甲基氯硅烷TMSCl，二氯甲烷CH2Cl2:四氢呋喃THF(体积比4:1)，0℃。
本发明采用13,14-双氯-β-榄香烯作为抗肿瘤活性骨架，具有如下重要优势：1)在β-榄香烯氯化反应中，会同时生成13-氯-β-榄香烯(中间体2，主产物)、14-氯-β-榄香烯(中间体3，副产物)和13,14-双氯-β-榄香烯(中间体4)，而两个单取代β-榄香烯氯代衍生物的极性极为相似，即便通过HPLC制备，也很难将二者分开，只能以二者的混合物为原料进行后续的反应，造成终产物中含有一定量的14位衍生物，不易纯化。因此，所制备得到的化合物难于纯化，不适合工业化大生产，不利于研发出可大规模制备的药物，因此难于进行药物的开发研究；2)制备13,14-双氯-β-榄香烯相对而言较易获得，且双氯代产物与单氯代β-榄香烯的极性差异较大，易于分离获得高纯度的双氯代β-榄香烯中间体，以及后续的衍生化终产物，且适宜于放大生产；3)13,14-双氯-β-榄香烯的抗肿瘤活性与13-氯-β-榄香烯、14-氯-β-榄香烯和β-榄香烯相当，且双氯代后轻微增加了化合物的极性，表现出了更好的类药性。
因此，本发明公开的β-榄香烯一氧化氮供体型衍生物易于制备分离，结构代谢稳定性更好，长期用药安全性更优，且可透过血脑屏障，强效地抑制恶性脑胶质瘤，具有很好的创新性和新颖性。
本发明提供了14-氯-β-榄香烯一氧化氮供体型衍生物，或其可药用盐，或溶剂化物，或其对映异构体、非对映异构体。
所述14-氯-β-榄香烯一氧化氮供体型衍生物具有如下通式(I)：
式(I)中：
R1为含氮和氧原子的直链或环状醇胺结构；
R2、R3分别独立选自Ci-10链烷基、C3-12环烷基、C6-12芳基、5-10元环杂芳基、C2-10烯基、C2-10炔基或C2-10烷氧基。
在一优选例中，式(I)中：
R1为C2-5的含氮和氧原子的直链醇胺结构或C5-6的环状醇胺结构；
R2、R3分别独立选自C2-6的链烷基、C2-6的烯基、C2-6的炔基。
在一优选例中，式(I)中：
R1选自
R2、R3分别独立选自-CH2CH2-、-CH2CH2CH2-、-CH2CH2CH2CH2-、-CH2CH2CH2CH2CH2-、-CH＝CH-、-CH2CH＝CH-、-CH＝CHCH2-、-CH2CH＝CHCH2-、-CH2CH2CH＝CH-、-CH＝CHCH2CH2-、-CH2C≡C-、-C≡CCH2-、-CH2C≡CCH2-、-CH2CH2C≡CCH2-、-CH2CH2C≡CCH2CH2-、-CH2C≡CCH2CH2-。
在一优选例中，所述14-氯-β-榄香烯一氧化氮供体型衍生物选自如下通式(I-1)～(I-6)：
式(I-1)～(I-6)中，R2、R3分别独立选自C2-6的链烷基、C2-6的烯基、C2-6的炔基，包括-CH2CH2-、-CH2CH2CH2-、-CH2CH2CH2CH2-、-CH2CH2CH2CH2CH2-、-CH＝CH-、-CH2CH＝CH-、-CH＝CHCH2-、-CH2CH＝CHCH2-、-CH2CH2CH＝CH-、-CH＝CHCH2CH2-、-CH2C≡C-、-C≡CCH2-、-CH2C≡CCH2-、-CH2CH2C≡CCH2-、-CH2CH2C≡CCH2CH2-、-CH2C≡CCH2CH2-等。
在一优选例中，所述14-氯-β-榄香烯一氧化氮供体型衍生物选自如下结构所示的化合物1～24：
本发明还提供了所述的14-氯-β-榄香烯一氧化氮供体型衍生物，或其可药用盐，或溶剂化物，或其对映异构体、非对映异构体在制备抗肿瘤药物中的应用。
所述肿瘤包括肺癌、结肠癌、恶性脑胶质瘤等。
本发明与现有技术相比，主要优点包括：
本发明引入了可提高体内抗肿瘤活性的醇胺结构作为连接臂，提高了体内稳定性，并且对临床上缺乏有效治疗药物的恶性脑胶质瘤具有很好的治疗活性。
附图说明
图1为本申请化合物与CN201710066664.1专利化合物相比的优势；
图2为13,14-双氯-β-榄香烯的合成路线；
图3为中间体5-10的合成路线；
图4为中间体12、13、14a、14b、15a、15b的合成路线；
图5为β-榄香烯一氧化氮供体型衍生物终产物Ia-d、IIa-b、IIIa-d、IVa-d、Va-b和VIa-b的合成路线；
图6为β-榄香烯和实施例4对脑恶性胶质瘤的抗肿瘤活性(n＝5)，A.体重，B.大脑重量，C.脑肿瘤的荧光信号强度，D.β-榄香烯和实施例4的抑制率；
图7中，A为模型组、β-榄香烯组和实施例4组的脑胶质瘤图像，B为脑胶质瘤肿组织的组织学分析。
具体实施方式
下面结合附图及具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的操作方法，通常按照常规条件，或按照制造厂商所建议的条件。
(一)中间体4的制备
如图2所示，在0℃条件下，向含β-榄香烯(1.02g，5mmol)的二氯甲烷(8mL)和四氢呋喃(2mL)混合溶液中，依次加入NCS(1.34g，20mmol)、三氟甲烷磺酸镱(310mg，0.5mmol)、三甲基氯硅烷(54mg，0.5mmol)，0℃下反应8h。反应结束时，减压蒸馏除去溶剂，加入水(15mL)稀释，用乙酸乙酯(4mL×3)萃取，合并有机相，依次用水(20mL×2)和饱和食盐水(20mL×2)洗涤，无水硫酸钠干燥，减压蒸馏除去溶剂，残留物经硅胶柱层析分离(纯石油醚)，得到无色液体化合物，即中间体4，产率为45％。
1H NMR(400MHz,CDCl3)δ5.85–5.72(m,1H),5.28(s,1H),5.18(s,1H),5.04(s,1H),4.98–4.89(m,3H),4.15–4.05(m,3H),3.97(d,J＝11.7Hz,1H),2.35–2.21(m,2H),1.77–1.42(m,6H),0.99(s,3H).13C NMR(100MHz,CDCl3)δ149.6,149.0,147.5,116.4,113.5,111.6,51.1,47.8,47.5,41.0,39.8,39.7,33.8,27.0,15.8.
(二)中间体5-10的制备
如图3所示，向含有中间体4(304mg，1.1mmol)的N,N-二甲基甲酰胺DMF(3mL)中，依次加入相应的醇胺反应物(1.3mmol)和N,N-二异丙基乙胺DIPEA(172mg，1.3mmol)，60℃搅拌12。反应结束时，向混合溶液加水(10mL)稀释，用乙酸乙酯(10mL)萃取，合并有机液，用水(10mL)和饱和食盐水(10mL)洗涤，无水硫酸钠干燥后，残留物经硅胶柱层析分离(二氯甲烷：甲醇体积比150：1)，得到淡黄色液体，即中间体5-10，图谱数据如下。
中间体5-10的1H NMR
2-((2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基))(甲基)氨基)乙-1-醇(5)
Light yellow liquid,yield 44％.1H NMR(400MHz,CDCl3)δ5.78(dd,J＝18.0,10.3Hz,1H),5.27(s,1H),5.01–4.88(m,5H),4.14–3.91(m,2H),3.61(t,J＝5.4Hz,2H),3.01(s,2H),2.56–2.51(m,2H),2.28(dd,J＝12.5,3.4Hz,1H),2.20(s,3H),2.11(t,J＝11.8Hz,1H),1.72–1.44(m,6H),0.98(s,3H).13C NMR(100MHz,CDCl3)δ151.0,149.3,147.8,116.2,111.4,111.3,63.1,59.0,58.6,51.0,41.9,41.7,40.0,39.9,34.1,27.3,16.0.HRMS(ESI)calcd for C18H31ClNO 312.2089[M+H]+,found 312.2080.
2-((2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)(异丙基)氨基)乙-1-醇(6)
Light yellow liquid,yield 60％.1H NMR(400MHz,CDCl3)δ5.77(dd,J＝17.1,11.2Hz,1H),5.26(s,1H),5.02–4.79(m,5H),4.02(dd,J＝47.3,11.7Hz,2H),3.51(s,2H),3.02(d,J＝18.7Hz,3H),2.54(s,2H),2.27(dd,J＝12.6,3.1Hz,1H),2.14–2.02(m,1H),1.71–1.41(m,6H),0.98(d,J＝7.2Hz,9H).13C NMR(100MHz,CDCl3)δ149.2,147.7,116.3,111.4,58.5,54.9,51.2,49.9,49.1,47.9,41.7,39.9,40.0,34.3,27.3,17.8,17.7,15.9.HRMS(ESI)calcd for C20H35ClNO 340.2402[M+H]+,found 340.2394.
(S)-1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)吡咯烷-3-醇(7)
Light yellow liquid,yield 48％.1H NMR(400MHz,CDCl3)δ5.77(dd,J＝17.1,11.1Hz,1H),5.26(s,1H),5.05–4.80(m,5H),4.30(s,1H),4.09(d,J＝11.6Hz,1H),3.96(d,J＝11.7Hz,1H),3.05(q,J＝13.4Hz,2H),2.85(td,J＝8.1,7.2,4.1Hz,1H),2.66(d,J＝10.2Hz,1H),2.55(s,1H),2.43(dd,J＝10.2,5.1Hz,1H),2.31–2.07(m,4H),1.69–1.41(m,6H),0.96(s,3H).13C NMR(100MHz,CDCl3)δ151.4,149.3,147.7,116.3,111.3,110.2,71.5,63.1,60.9,52.6,51.1,47.7,42.0,39.9,39.8,35.0,34.0,27.1,15.8.HRMS(ESI)calcdfor C19H31ClNO 324.2089[M+H]+,found 324.2083.
(1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌啶-4-基)甲醇(8)
Light yellow liquid,yield 60％.1H NMR(400MHz,CDCl3)δ5.79(dd,J＝17.9,10.4Hz,1H),5.26(s,1H),4.98–4.86(m,5H),4.09(d,J＝11.7Hz,1H),3.97(d,J＝11.7Hz,1H),3.50(d,J＝6.4Hz,2H),2.90(d,J＝14.5Hz,4H),2.32–2.23(m,1H),2.13(d,J＝7.9Hz,1H),1.88(s,2H),1.75–1.57(m,7H),1.53–1.43(m,4H),0.98(s,3H).13C NMR(100MHz,CDCl3)δ149.1,147.6,115.9,110.9,67.8,63.5,53.5,53.3,50.8,47.6,42.0,39.7,39.6,38.5,33.8,28.6,26.8,15.6.HRMS(ESI)calcd for C21H35ClNO 352.2402[M+H]+,found352.2384.
(1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌啶-3-基)甲醇(9)
Light yellow liquid,yield 51％.1H NMR(400MHz,CDCl3)δ5.83–5.73(m,1H),5.26(s,1H),4.91(dd,J＝10.5,6.4Hz,5H),4.14–4.02(m,1H),3.97(d,J＝11.7Hz,1H),3.69–3.54(m,2H),2.89(d,J＝3.9Hz,2H),2.68(d,J＝9.7Hz,1H),2.47(s,2H),2.27(dd,J＝11.0,5.0Hz,1H),2.09(ddt,J＝12.2,8.2,4.3Hz,2H),1.82–1.74(m,2H),1.70–1.41(m,9H),0.97(s,3H).13C NMR(100MHz,CDCl3)δ150.7,149.4,147.8,147.8,116.2,111.3,111.0,67.5,64.1,57.8,54.4,51.0,47.8,42.1,39.9,39.9,34.0,27.8,27.0,24.6,15.8.HRMS(ESI)calcd for C21H35ClNO 352.2402[M+H]+,found 352.2412.
2-((4-(2-(4-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌嗪-1-基)乙-1-醇(10)
Light yellow liquid,yield 44％.1H NMR(400MHz,CDCl3)δ5.78(dd,J＝18.0,10.2Hz,1H),5.26(s,1H),4.97–4.85(m,5H),4.09(d,J＝11.5Hz,1H),3.96(d,J＝11.6Hz,1H),3.61(t,J＝5.4Hz,2H),2.91(d,J＝4.2Hz,2H),2.62–2.38(m,10H),2.30–2.25(m,1H),2.15–2.08(m,1H),1.64–1.43(m,6H),0.97(s,3H).13C NMR(100MHz,CDCl3)δ150.7,149.4,147.8,116.3,111.3,111.0,63.5,59.4,57.8,53.3,53.1,51.2,47.7,42.2,39.9,39.9,34.0,27.1,15.8.HRMS(ESI)calcd for C21H36ClN2O 367.2511[M+H]+,found367.2503.
(三)中间体12、13的制备
在0℃条件下，向含有3,4-双(苯磺酰基)-1,2,5-恶二唑2-氧化物(183mg，0.5mmol)的四氢呋喃溶液中，缓慢滴加25％NaOH水溶液(0.2mL)，滴加完毕后，继续搅拌10min，加入反应物乙二醇(124mg，2mmol)，室温条件下反应6h。减压蒸馏除去溶剂，加入水(15mL)稀释，用二氯甲烷(20mL×3)萃取，合并有机相，依次用水(20mL×2)和饱和食盐水(20mL×2)洗涤，无水硫酸钠干燥，减压蒸馏除去溶剂，残留物经硅胶层析(二氯甲烷：甲醇体积比400：1)得到中间体12(72mg，48％)。
m.p.128-130℃.1H NMR(400MHz,CDCl3)δ8.07(d,J＝8.2Hz,2H),7.77(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),4.59–4.49(m,2H),4.08–4.02(m,2H).13C NMR(100MHz,CDCl3)δ159.1,137.8,135.9,129.9,128.7,110.7,73.0,60.5.HRMS(ESI)calcd forC10H10N2NaO6S 309.0152[M+Na]+,found 309.0142.
按照图4和上述相同的方法，更换反应物(将乙二醇换成1,4丁炔二醇)，制得中间体13。
中间体13的1H NMR
4-((4-羟基丁基2-炔-1-基)氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物(13)
White solid,yield 53％,m.p.116-118℃.1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.7Hz,2H),7.76(t,J＝7.4Hz,1H),7.63(t,J＝7.7Hz,2H),5.10(s,2H),4.34(s,2H).13CNMR(100MHz,CDCl3)δ158.2,138.0,135.9,129.8,128.8,110.8,88.3,77.7,59.0,51.1.HRMS(ESI)calcd for C12H10N2NaO6S 333.0512[M+Na]+,found 333.0512.
(四)中间体14a、14b、15a、15b的制备
如图4所示，向含有中间体12(182mg，0.63mmol)的二氯甲烷(5mL)溶液中，依次加入反应物丁二酸酐(76mg，0.76mmol)和4-二甲氨基吡啶DMAP(38mg，0.32mmol)，室温搅拌6h，在室温条件下，向混合溶液加入水(5mL)淬灭，继续搅拌10min，用二氯甲烷(5mL×3)萃取，合并有机相，依次用水(20mL×2)和饱和食盐水(20mL×2)洗涤，无水硫酸钠干燥，减压蒸馏除去溶剂，残留物经硅胶柱层析分离(二氯甲烷：甲醇体积比150：1)得到白色固体，即中间体14a(167mg，67％)。
m.p.118-120℃.1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.3Hz,2H),7.76(t,J＝7.5Hz,1H),7.63(t,J＝7.9Hz,2H),4.72–4.59(m,2H),4.57–4.44(m,2H),2.92–2.51(m,4H).13C NMR(100MHz,CDCl3)δ176.9,171.9,158.8,138.2,135.8,129.8,128.8,110.6,69.0,61.6,29.0,28.9.HRMS(ESI)calcd for C14H14N2NaO9S 409.0312[M+Na]+,found409.0295.
按照图4和上述相同的方法，更换反应物，制得中间体14b和15a、15b。
中间体14b和15a、15b的1H NMR
4-(2-((4-羧基丁酰基)氧基)乙氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物(14b)
White solid,yield 53％,m.p.91-93℃.1H NMR(400MHz,CDCl3)δ8.06(d,J＝8.1Hz,2H),7.76(t,J＝7.5Hz,1H),7.62(t,J＝7.8Hz,2H),4.66–4.60(m,2H),4.55–4.48(m,2H),2.47(q,J＝7.0Hz,4H),1.98(p,J＝7.2Hz,2H).13C NMR(100MHz,CDCl3)δ178.6,172.7,158.8,138.0,135.8,129.8,128.8,110.6,69.0,61.3,33.0,32.9,19.7.HRMS(ESI)calcd for C15H16N2NaO9S 423.0469[M+Na]+,found 423.0465.
4-((4-((3-羧基丙酰基)氧基)丁-2-炔-1-基)氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物(15a)
White solid,yield 54％,m.p.106-108℃.1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.4Hz,2H),7.77(t,J＝7.5Hz,1H),7.63(t,J＝7.9Hz,2H),5.10(s,2H),4.77(s,2H),2.70(dt,J＝8.1,4.7Hz,4H).13C NMR(100MHz,CDCl3)δ177.2,171.4,158.1,138.1,135.9,129.9,128.8,110.8,83.9,78.8,58.8,52.3,28.8,28.8.HRMS(ESI)calcd forC16H14N2NaO9S 433.0312[M+Na]+,found 433.0317.
4-(4-((4-羧基丁酰)氧基)丁-2-炔-1-基氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物(15b)
White solid,yield 54％,m.p.94-96℃.1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.6Hz,2H),7.77(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.10(s,2H),4.74(s,2H),2.46(td,J＝7.3,4.9Hz,4H),1.97(p,J＝7.3Hz,2H).13C NMR(100MHz,CDCl3)δ179.0,172.1,158.0,137.7,135.8,129.8,128.7,110.6,83.9,78.6,58.6,52.0,32.8,19.6.HRMS(ESI)calcd for C16H14N2NaO9S 447.0469[M+Na]+,found 447.0486.
(五)β-榄香烯一氧化氮供体型衍生物终产物Ia-Id、IIa-IIb、IIIa-IIId、IVa-IVd、Va-Vb和VIa-VIb的制备
如图5所示，将中间体5-10(0.08mmol)、NO供体中间体14a、14b和15a、15b(0.10mmol)、1-乙基-(3-二甲基氨基丙基)碳二亚胺盐酸盐EDCI(0.12mmol)和DMAP(0.01mmol)在无水二氯甲烷(2mL)中在室温下搅拌8小时。采用薄层色谱法检测反应。用二氯甲烷(5mL)稀释反应混合物。有机层依次用水和盐水洗涤，无水硫酸钠干燥，减压蒸馏除去溶剂，残留物通过柱色层析(二氯甲烷/甲醇)纯化，使产物为浅黄色液体。
所采用的各中间体均可按上述(一)～(四)进行制备，在此不再赘述。
具体的：
实施例1
4-(2-((4-(2-((2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基))(甲基)氨基)乙氧基)-4-氧代丁酰基)氧基)乙氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备。
向含有中间体5(26mg，0.084mmol)的二氯甲烷(1.5mL)溶液中，依次加入中间体14a(39mg，0.101mmol)、DMAP(1mg，0.008mmol)、EDCI(24mg，0.126mmol)，室温搅拌8h。加二氯甲烷(5mL)稀释，依次用水(10mL×2)和饱和食盐水(10mL×2)洗涤，无水硫酸钠干燥，减压蒸馏除去溶剂，残留物经硅胶柱层析分离(二氯甲烷：甲醇体积比400：1)，淡黄色液体，产率69％。
1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.8Hz,2H),7.75(d,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.78(dd,J＝17.8,10.4Hz,1H),5.26(s,1H),5.02–4.87(m,5H),4.65–4.59(m,2H),4.57–4.48(m,2H),4.21(t,J＝5.8Hz,2H),4.09(d,J＝11.7Hz,1H),3.96(d,J＝11.8Hz,1H),3.01(s,2H),2.63(d,J＝5.5Hz,6H),2.32–2.22(m,4H),1.66–1.42(m,6H),0.97(s,3H).13C NMR(100MHz,CDCl3)δ172.0,172.0,158.7,149.2,147.7,138.1,135.6,129.7,128.6,116.1,111.2,110.4,77.0,68.9,62.5,61.4,55.3,50.8,47.9,42.6,41.5,39.8,33.8,29.0,28.9,27.1,15.8.HRMS(ESI)calcd for C32H43ClN3O9S 680.2403[M+H]+,found 680.2441.
实施例2
4-(2-((5-(2-((2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)(甲基)氨基)乙氧基)-5-氧戊烷酰基)氧基)乙氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例1，区别在于，区别在于，将(三)中的丁二酸酐替换为戊二酸酐。
本实施例制得黄色蜡状液体，产率71％。
1H NMR(400MHz,CDCl3)δ8.06(d,J＝8.5Hz,2H),7.76(t,J＝7.5Hz,1H),7.63(t,J＝7.9Hz,2H),5.87–5.71(m,1H),5.26(s,1H),5.01–4.86(m,5H),4.67–4.60(m,2H),4.53–4.46(m,2H),4.17(t,J＝5.7Hz,2H),4.09(d,J＝11.6Hz,1H),3.96(d,J＝11.7Hz,1H),2.95(s,2H),2.57(t,J＝5.1Hz,2H),2.43(dt,J＝17.0,7.3Hz,4H),2.28(dd,J＝11.9,4.0Hz,1H),2.16–2.07(m,1H),2.02–1.93(m,2H),1.65–1.35(m,6H),0.96(s,3H).13CNMR(100MHz,CDCl3)δ173.0,172.8,158.8,151.4,149.4,147.8,138.1,135.8,129.8,128.7,116.3,110.9,110.5,69.0,63.3,62.5,61.2,55.4,51.1,47.7,42.9,41.5,39.9,39.9,33.9,33.2,33.1,27.1,20.0,15.8.HRMS(ESI)calcd for C33H45ClN3O9S 694.2560[M+H]+,found694.2562.
实施例3
4-((4-((4-(2-((2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)(甲基)氨基)乙氧基)-4-氧代丁酰基)氧基)丁-2-炔-1-基)氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例1，区别在于，将(三)中的乙二醇替换为1,4-丁炔二醇。
本实施例制得黄色蜡状液体，产率74％。
1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.4Hz,2H),7.77(t,J＝7.5Hz,1H),7.64(t,J＝7.9Hz,2H),5.78(dd,J＝16.9,11.3Hz,1H),5.26(s,1H),5.10(s,2H),5.05–4.85(m,5H),4.76(s,2H),4.20(t,J＝5.6Hz,2H),4.12–4.06(m,1H),3.96(d,J＝11.7Hz,1H),2.97(s,2H),2.70–2.56(m,6H),2.31–2.09(m,5H),1.70–1.41(m,6H),0.97(s,3H).13C NMR(100MHz,CDCl3)δ172.2,171.6,158.0,151.3,149.4,149.4,147.7,137.9,135.9,129.9,128.8,116.3,111.3,110.7,83.9,78.7,63.2,62.8,58.7,55.3,52.3,51.1,47.7,42.8,41.4,39.9,39.9,33.9,29.0,28.9,27.1,15.8.HRMS(ESI)calcd for C34H43ClN3O9S704.2403[M+H]+,found 704.2408.
实施例4
4-((4-((5-(2-((2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)(甲基)氨基)乙氧基)-5-氧戊烷氧基)丁-2-炔-1-基氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例3，区别在于，将(三)中的丁二酸酐换成戊二酸酐。
本实施例制得黄色蜡状液体，产率73％。
1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.4Hz,2H),7.77(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.77(dd,J＝17.1,11.2Hz,1H),5.25(s,1H),5.09(s,2H),4.98–4.83(m,5H),4.73(s,2H),4.17(t,J＝5.8Hz,2H),4.09(d,J＝11.7Hz,1H),3.96(d,J＝11.7Hz,1H),2.96(s,2H),2.57(t,J＝5.5Hz,2H),2.41(dt,J＝17.1,7.3Hz,4H),2.31–2.18(m,4H),2.12(s,1H),1.96(p,J＝7.1Hz,2H),1.68–1.41(m,6H),0.96(s,3H).13C NMR(100MHz,CDCl3)δ172.9,172.2,158.0,151.3,149.3,147.7,137.8,135.9,129.8,128.8,116.3,111.3,110.9,110.7,84.0,78.6,63.2,62.4,58.7,55.4,52.0,51.1,47.7,39.9,33.8,33.2,33.0,27.1,20.0,15.8.HRMS(ESI)calcd for C35H45ClN3O9S 718.2560[M+H]+,found718.2553.
实施例5
4-(2-((4-(2-((2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)(异丙基)氨基)乙氧基)-4-氧代丁酰基)氧基)乙氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例1，区别在于，将(一)中的N-甲基-2-羟基乙胺换成2-(异丙基氨)乙醇。
本实施例中制得黄色蜡状液体，产率58％。
1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.5Hz,2H),7.76(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.77(dd,J＝17.6,10.6Hz,1H),5.25(s,1H),5.03–4.84(m,5H),4.66–4.58(m,2H),4.56–4.47(m,2H),4.11–3.93(m,4H),3.06–2.82(m,3H),2.72–2.56(m,6H),2.30–2.09(m,2H),1.65–1.40(m,6H),1.02–0.90(m,9H).13C NMR(100MHz,CDCl3)δ172.2,172.1,158.7,149.8,149.3,147.7,138.0,135.7,129.7,128.7,116.2,111.2,110.4,110.0,68.9,64.0,61.4,55.6,51.2,50.2,47.7,41.3,39.9,39.9,34.0,28.9,28.9,27.1,18.0,17.7,15.7.HRMS(ESI)calcd for C34H47ClN3O9S 708.2716[M+H]+,found708.2703.
实施例6
4-(2-((5-(2-((2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)(异丙基)氨基)乙氧基)-5-氧戊烷氧基)乙氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例2，区别在于，将(一)中的N-甲基-2-羟基乙胺换成2-(异丙基氨)乙醇。
本实施例中制得黄色蜡状液体，产率68％。
1H NMR(400MHz,CDCl3)δ8.06(d,J＝7.4Hz,2H),7.76(t,J＝7.5Hz,1H),7.62(t,J＝7.8Hz,2H),5.77(dd,J＝16.9,11.3Hz,1H),5.25(s,1H),5.03–4.82(m,5H),4.65–4.56(m,2H),4.54–4.43(m,2H),4.12–3.91(m,4H),3.08–2.79(m,3H),2.58(t,J＝6.6Hz,2H),2.42(dt,J＝26.0,7.3Hz,4H),2.26(dd,J＝12.7,3.4Hz,1H),2.13(t,J＝11.3Hz,1H),1.97(p,J＝7.3Hz,2H),1.66–1.41(m,6H),1.01–0.87(m,9H).13C NMR(100MHz,CDCl3)δ172.9,172.8,158.8,149.8,149.3,147.8,138.0,135.8,129.8,128.7,116.3,111.3,110.5,110.1,69.0,63.8,61.2,55.6,51.2,50.2,47.8,41.4,40.0,39.9,34.1,33.2,33.1,27.1,20.0,18.1,17.8,15.8.HRMS(ESI)calcd for C35H48ClN3O9S 722.2873[M+H]+,found722.2873.
实施例7
4-((4-((4-(2-((2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)(异丙基)氨基)乙氧基)-4-氧代丁酰基)氧基)丁-2-炔-1-基)氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例3，区别在于，将(一)中的N-甲基-2-羟基乙胺换成2-(异丙基氨)乙醇。
本实施例中制得黄色蜡状液体，产率47％。
1H NMR(400MHz,CDCl3)δ8.08(d,J＝7.7Hz,2H),7.77(t,J＝7.4Hz,1H),7.64(t,J＝7.8Hz,2H),5.78(dd,J＝17.1,11.1Hz,1H),5.25(s,1H),5.10(s,2H),4.76(s,2H),4.18–3.91(m,4H),2.98(d,J＝45.6Hz,3H),2.65(dt,J＝10.6,5.1Hz,6H),2.31–2.11(m,2H),1.64–1.41(m,6H),0.97(s,9H).13C NMR(100MHz,CDCl3)δ172.1,171.6,158.1,149.4,147.9,138.1,135.8,129.9,128.8,116.2,111.3,110.7,84.0,78.8,58.7,55.7,52.2,51.1,48.0,47.9,41.5,40.0,34.2,29.1,29.0,27.3,22.8,18.1,17.8,15.9.HRMS(ESI)calcd for C36H47ClN3O9S 732.2716[M+H]+,found 732.2711.
实施例8
4-((4-((5-(2-((2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)(异丙基氨基)乙氧基)-5-氧戊烷酰基)氧基)丁-2-炔-1-基)氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例4，区别在于，将(一)中的步骤1中的N-甲基-2-羟基乙胺换成2-(异丙基氨)乙醇。
本实施例中制得黄色蜡状液体，产率53％。
1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.4Hz,2H),7.77(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.78(dd,J＝17.0,11.2Hz,1H),5.25(s,1H),5.09(s,2H),5.04–4.81(m,5H),4.74(s,2H),4.13–3.91(m,4H),3.09–2.79(m,3H),2.59(t,J＝6.2Hz,2H),2.40(dt,J＝26.3,7.4Hz,5H),2.26(dd,J＝12.2,3.5Hz,1H),2.14(s,1H),1.96(p,J＝7.3Hz,2H),1.66–1.41(m,6H),1.06–0.89(m,9H).13C NMR(100MHz,CDCl3)δ172.7,172.0,157.9,149.3,147.7,137.9,135.7,129.7,128.7,116.1,111.1,110.6,110.0,84.0,78.5,63.7,58.6,55.6,51.8,51.0,50.3,47.9,47.9,41.4,39.9,39.9,34.1,33.2,33.0,27.1,20.0,18.0,17.7,15.8.HRMS(ESI)calcd for C37H49ClN3O9S 746.2873[M+H]+,found 746.2871.
实施例9
4-(2-((4-(((S)-1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)吡咯烷-3-基)氧基)-4-氧代丁酰基)氧基)乙氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例1，区别在于，将(一)中的N-甲基-2-羟基乙胺换成(R)-3-吡咯烷醇。
本实施例中制得黄色蜡状液体，产率68％。
1H NMR(400MHz,CDCl3)δ8.06(d,J＝7.5Hz,2H),7.76(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.77(dd,J＝17.1,11.2Hz,1H),5.26(s,1H),5.17(s,1H),5.00–4.83(m,5H),4.66–4.57(m,2H),4.56–4.48(m,2H),4.09(d,J＝11.6Hz,1H),3.96(d,J＝11.7Hz,1H),3.12(d,J＝13.4Hz,1H),2.99(d,J＝13.4Hz,1H),2.76–2.60(m,7H),2.40(s,1H),2.31–2.19(m,2H),1.87–1.79(m,1H),1.67–1.40(m,6H),0.96(s,3H).13C NMR(100MHz,CDCl3)δ172.2,172.2,158.8,149.3,147.7,138.0,135.8,129.8,128.7,116.2,111.3,110.5,74.7,68.9,61.5,60.8,60.0,53.0,51.1,47.6,42.0,39.9,39.8,34.0,31.9,29.2,29.0,27.1,15.8.HRMS(ESI)calcd for C33H43ClN3O9S 692.2403[M+H]+,found692.2394.
实施例10
4-(2-((5-(((S)-1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)吡咯烷-3-基)氧基)-5-氧戊烷酰基)氧基)乙氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例2，区别在于，将(一)中的N-甲基-2-羟基乙胺换成(R)-3-吡咯烷醇。
本实施例中制得黄色蜡状液体，产率62％。
1H NMR(400MHz,CDCl3)δ8.05(d,J＝7.5Hz,2H),7.76(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.77(dd,J＝17.0,11.2Hz,1H),5.25(s,1H),5.16(s,1H),5.03–4.85(m,5H),4.66–4.59(m,2H),4.53–4.46(m,2H),4.09(d,J＝11.6Hz,1H),3.96(d,J＝11.7Hz,1H),3.17–2.91(m,2H),2.71(d,J＝6.6Hz,2H),2.59(d,J＝9.8Hz,1H),2.42(dt,J＝23.1,7.3Hz,5H),2.31–2.19(m,2H),2.11(s,1H),2.00–1.92(m,2H),1.85–1.76(m,1H),1.67–1.40(m,6H),0.96(s,3H).13C NMR(100MHz,CDCl3)δ172.9,172.8,158.8,149.3,147.7,138.0,135.8,129.8,128.7,116.2,111.3,110.5,74.4,69.0,61.2,60.9,60.0,53.0,51.1,47.6,42.0,39.9,39.8,34.0,33.4,33.1,32.0,27.1,20.0,15.8.HRMS(ESI)calcd forC34H45ClN3O9S 706.2560[M+H]+,found 706.2555.
实施例11
4-((4-((4-(((S)-1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)吡咯烷-3-基)氧基)-4-氧代丁酰基)氧基)丁-2-炔-1-基)氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实例3，区别在于，将(一)中的N-甲基-2-羟基乙胺换成(R)-3-吡咯烷醇。
本实施例中制得黄色蜡状液体，产率53％。
1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.4Hz,2H),7.77(t,J＝7.5 8Hz,1H),7.63(t,J＝7.9Hz,2H),5.78(dd,J＝17.9,10.3Hz,1H),5.26(s,1H),5.19(s,1H),5.09(s,2H),5.02–4.84(m,5H),4.76(s,2H),4.09(d,J＝11.4Hz,1H),3.97(d,J＝11.8Hz,1H),3.16–2.95(m,2H),2.76–2.60(m,7H),2.41(s,1H),2.31–2.08(m,3H),1.88–1.79(m,1H),1.68–1.43(m,6H),0.97(s,3H).13C NMR(100MHz,CDCl3)δ172.0,171.6,158.1,149.4,147.8,147.8,138.0,135.8,129.8,128.8,116.2,111.3,110.7,84.0,78.8,74.8,60.9,60.0,58.7,53.0,52.2,51.0,47.9,42.2,39.9,39.9,34.1,32.0,29.2,29.0,27.2,15.9.HRMS(ESI)calcd for C35H43ClN3O9S 716.2403[M+H]+,found 716.2400.
实施例12
4-((4-((5-(((S)-1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)吡咯烷-3-基)氧基)-5-氧戊烷酰基)氧基)丁-2-炔-1-基)氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实例4，区别在于，将(一)中的N-甲基-2-羟基乙胺换成(R)-3-吡咯烷醇。
本实施例中制得黄色蜡状液体，产率63％。
1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.4Hz,2H),7.76(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.78(dd,J＝17.9,10.3Hz,1H),5.26(s,1H),5.17(td,J＝6.2,3.2Hz,1H),5.09(s,2H),5.01–4.82(m,5H),4.74(s,2H),4.09(d,J＝11.5Hz,1H),3.96(d,J＝11.8Hz,1H),3.11(d,J＝13.4Hz,1H),2.99(d,J＝13.4Hz,1H),2.78–2.66(m,2H),2.60(d,J＝10.9Hz,1H),2.40(dt,J＝23.9,7.3Hz,5H),2.31–2.17(m,2H),2.10(dq,J＝7.6,5.3,3.5Hz,1H),1.95(p,J＝7.3Hz,2H),1.81(dq,J＝13.7,8.7,6.9Hz,1H),1.70–1.39(m,6H),0.97(s,3H).13C NMR(100MHz,CDCl3)δ172.8,172.2,158.0,151.1,149.3,147.7,137.8,135.9,129.8,128.7,116.2,111.3,110.7,110.4,84.0,78.6,74.4,60.8,60.0,58.7,53.0,52.0,51.0,47.6,42.0,39.9,39.8,34.0,33.3,33.0,31.9,27.1,20.0,15.8.HRMS(ESI)calcd for C36H45ClN3O9S 730.2560[M+H]+,found 730.255.
实施例13
4-(2-((4-((1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌啶-4-基)甲氧基)-4-氧代丁酰基)氧基)乙氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例1，区别在于，将(一)中的N-甲基-2-羟基乙胺替换成4-羟甲基哌啶。
本实施例中制得黄色蜡状液体，产率67％。
1H NMR(400MHz,CDCl3)δ8.06(d,J＝7.7Hz,2H),7.76(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.78(dd,J＝17.1,11.1Hz,1H),5.25(s,1H),4.97–4.84(m,5H),4.66–4.56(m,2H),4.57–4.47(m,2H),4.13–3.88(m,3H),2.88(d,J＝17.1Hz,4H),2.68(s,4H),2.34–2.20(m,1H),2.12(s,1H),1.85(s,2H),1.70–1.38(m,10H),0.96(s,3H).13C NMR(100MHz,CDCl3)δ172.25,172.15,158.69,149.29,147.72,137.94,135.74,129.73,128.66,116.14,111.18,110.43,69.29,68.86,63.64,61.42,53.42,53.28,51.08,47.62,42.10,39.85,39.74,35.34,33.91,28.94,28.92,26.95,15.69.HRMS(ESI)calcd for C35H47ClN3O9S720.2716[M+H]+,found 720.2709.
实施例14
4-(2-((5-((1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌啶-4-基)甲氧基)-5-氧戊烷酰基)氧基)乙氧基)-3-(苯磺酰基)-1,2,5-恶二唑2-氧化物的制备方法基本同实施例2，区别在于，将(一)中的N-甲基-2-羟基乙胺替换成4-羟甲基哌啶。
本实施例中制得黄色蜡状液体，产率70％。
1H NMR(400MHz,CDCl3)δ8.06(d,J＝7.4Hz,2H),7.76(t,J＝7.5Hz,1H),7.62(t,J＝7.9Hz,2H),5.78(dd,J＝18.0,10.3Hz,1H),5.26(s,1H),5.01–4.85(m,5H),4.72–4.59(m,2H),4.54–4.46(m,2H),4.08(d,J＝11.7Hz,1H),3.98–3.90(m,3H),3.03–2.80(m,4H),2.42(dt,J＝18.4,7.3Hz,4H),2.31–2.23(m,1H),2.15–2.07(m,1H),2.03–1.86(m,4H),1.68–1.42(m,10H),0.97(s,3H).13C NMR(100MHz,CDCl3)δ173.0,172.8,158.8,149.4,147.8,138.1,135.8,129.8,128.7,116.2,111.3,110.5,69.0,69.0,63.6,61.2,53.4,53.3,51.1,47.8,42.2,39.9,39.8,35.4,34.0,33.3,33.2,29.8,28.8,27.1,20.1,15.8.HRMS(ESI)calcd for C36H49ClN3O9S 734.2873[M+H]+,found 734.889.
实施例15
4-((4-((4-((1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌啶-4-基)甲氧基)-4-氧代丁酰基)氧基)丁-2-炔-1-基)氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例3，区别在于，将(一)中的N-甲基-2-羟基乙胺替换成4-羟甲基哌啶。
本实施例中制得黄色蜡状液体，产率65％。
1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.4Hz,2H),7.76(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.78(dd,J＝17.9,10.4Hz,1H),5.26(s,1H),5.09(s,2H),5.02–4.85(m,5H),4.76(s,2H),4.08(d,J＝11.4Hz,1H),3.99–3.94(m,3H),2.98(s,4H),2.66(q,J＝3.7Hz,4H),2.30–2.25(m,1H),2.17–2.11(m,1H),1.97(d,J＝17.5Hz,2H),1.71–1.43(m,10H),0.97(s,3H).13C NMR(100MHz,CDCl3)δ172.1,171.6,158.1,149.3,147.9,138.1,135.8,129.8,128.8,116.1,111.3,110.7,84.0,78.8,69.1,65.7,63.5,58.7,53.4,53.3,52.2,50.9,48.0,42.3,39.9,39.9,35.3,34.1,29.8,29.1,29.0,27.2,15.9.HRMS(ESI)calcdfor C37H47ClN3O9S 744.2716[M+H]+,found 744.2700.
实施例16
4-((4-((5-((1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌啶-4-基)甲氧基)-5-氧戊烷酰基)氧基)丁-2-炔-1-基)氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例4，区别在于，将(一)中的N-甲基-2-羟基乙胺替换成4-羟甲基哌啶。
本实施例中制得黄色蜡状液体，产率60％。
1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.5Hz,2H),7.77(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.78(dd,J＝17.5,10.8Hz,1H),5.25(s,1H),5.09(s,2H),5.00–4.85(m,5H),4.74(s,2H),4.09(d,J＝11.6Hz,1H),4.01–3.87(m,3H),2.89(d,J＝17.0Hz,4H),2.41(dt,J＝18.4,7.3Hz,5H),2.30–2.23(m,1H),2.16–2.06(m,1H),2.04–1.77(m,5H),1.73–1.39(m,10H),0.96(s,3H).13C NMR(100MHz,CDCl3)δ173.0,172.2,158.0,149.4,147.8,137.8,135.9,129.8,128.8,116.2,111.3,110.7,84.0,78.6,76.8,69.1,63.7,58.7,53.5,53.4,52.0,51.1,47.7,42.2,39.9,39.8,35.4,34.0,33.2,33.0,29.0,27.0,20.1,15.8.HRMS(ESI)calcd for C38H49ClN3O9S 758.2873[M+H]+,found 758.2866.
实施例17
4-(2-((4-((1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌啶-3-基)甲氧基)-4-氧代丁酰基)氧基)乙氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例1，区别在于，将(一)中的N-甲基-2-羟基乙胺替换成3-羟甲基哌啶。
本实施例中制得黄色蜡状液体，产率70％。
1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.4Hz,2H),7.77(t,J＝7.5Hz,1H),7.63(t,J＝7.9Hz,2H),5.78(dd,J＝17.9,10.3Hz,1H),5.26(s,1H),4.98–4.82(m,5H),4.65–4.60(m,2H),4.54–4.50(m,2H),4.14–3.87(m,4H),2.88(s,2H),2.67(t,J＝3.8Hz,6H),2.32–2.23(m,1H),2.11(s,1H),1.91(s,2H),1.78–1.41(m,12H),0.97(s,3H).13CNMR(100MHz,CDCl3)δ172.3,172.2,158.8,151.1,149.4,147.8,138.1,135.8,129.8,128.8,116.2,111.3,111.3,110.5,69.0,67.7,64.0,61.5,57.0,54.2,51.1,47.8,42.2,39.9,39.9,35.7,34.1,34.0,29.0,27.3,27.1,24.6,15.8.HRMS(ESI)calcd forC35H47ClN3O9S720.2716[M+H]+,found 720.2721.
实施例18
4-(2-((5-((1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌啶-3-基)甲氧基)-5-氧戊烷酰基)氧基)乙氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例2，区别在于，将(一)中的N-甲基-2-羟基乙胺替换成3-羟甲基哌啶。
本实施例中制得黄色蜡状液体，产率74％。
1H NMR(400MHz,CDCl3)δ8.06(d,J＝7.5Hz,2H),7.76(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.77(dd,J＝17.3,10.9Hz,1H),5.25(s,1H),4.98–4.80(m,5H),4.68–4.57(m,2H),4.55–4.45(m,2H),4.09(d,J＝11.6Hz,1H),3.94(dd,J＝21.4,11.3Hz,3H),2.99–2.54(m,4H),2.42(dt,J＝20.6,7.3Hz,4H),2.30–2.23(m,1H),2.11(s,1H),1.97(p,J＝7.3Hz,4H),1.76–1.36(m,10H),0.97(s,3H).13C NMR(100MHz,CDCl3)δ173.0,172.8,158.8,149.4,147.8,138.0,135.8,129.8,128.7,116.2,111.3,110.5,69.0,67.3,63.9,61.2,57.0,54.2,51.1,47.8,42.2,39.9,39.8,35.7,34.0,34.0,33.2,33.1,27.3,27.0,24.6,20.0,15.8.HRMS(ESI)calcd for C36H49ClN3O9S 734.2873[M+H]+,found734.2871.
实施例19
4-((4-((4-((1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌啶-3-基)甲氧基)-4-氧代丁酰基)氧基)丁-2炔-1-基)氧基)-3-(苯磺酰基)-1,2,5-恶二唑2-氧化物的制备方法基本同实施例3，区别在于，将(一)中的N-甲基-2-羟基乙胺替换成3-羟甲基哌啶。
本实施例中制得黄色蜡状液体，产率70％。
1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.5Hz,2H),7.77(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.78(dd,J＝17.8,10.4Hz,1H),5.25(s,1H),5.10(s,2H),4.91(d,J＝14.0Hz,5H),4.76(s,2H),4.11–3.91(m,4H),2.88(s,2H),2.80–2.51(m,J＝3.7Hz,7H),2.31–2.21(m,1H),2.12(s,1H),1.92(s,3H),1.71–1.39(m,10H),0.97(s,3H).13C NMR(100MHz,CDCl3)δ172.2,171.6,158.0,149.4,147.8,137.9,135.9,129.8,128.8,116.2,111.3,110.7,83.9,78.7,67.7,63.9,58.7,57.0,54.2,52.2,51.1,47.8,42.2,39.9,39.8,35.7,34.0,29.0,28.9,27.3,27.0,24.6,15.8.HRMS(ESI)calcd forC37H47ClN3O9S744.2716[M+H]+,found 744.2709.
实施例20
4-((4-((5-((1-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌啶-3-基)甲氧基)-5-氧戊烷酰基)氧基)丁-2-炔-1-基)氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例4，区别在于，将(一)中的N-甲基-2-羟基乙胺替换成3-羟甲基哌啶。
本实施例中制得黄色蜡状液体，产率73％。
1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.7Hz,2H),7.77(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.78(dd,J＝17.9,10.4Hz,1H),5.26(s,1H),5.10(s,2H),4.92(d,J＝14.0Hz,5H),4.74(s,2H),4.09(d,J＝11.7Hz,1H),3.94(dd,J＝18.4,11.3Hz,3H),3.02–2.57(m,5H),2.40(dt,J＝20.6,7.3Hz,5H),2.31–2.23(m,1H),2.11(s,1H),1.96(p,J＝7.3Hz,4H),1.70–1.42(m,10H),0.97(s,3H).13C NMR(100MHz,CDCl3)δ172.9,172.2,158.0,151.1,149.4,147.8,138.0,135.9,129.8,128.8,116.2,111.3,110.7,84.1,78.7,67.4,64.0,58.7,57.0,54.2,52.0,51.1,47.9,42.3,39.9,39.9,35.8,34.1,33.2,33.1,27.3,27.1,24.7,20.1,15.9.HRMS(ESI)calcd for C38H49ClN3O9S 758.2873[M+H]+,found758.2897.
实施例21
4-(2-((4-(2-(4-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌嗪-1-基)乙氧基)-4-氧代丁酰基)氧基)乙氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例1，区别在于，将(一)中的N-甲基-2-羟基乙胺替换成N-羟乙基哌嗪。
本实施例中制得黄色蜡状液体，产率53％。
1H NMR(400MHz,CDCl3)δ8.06(d,J＝7.3Hz,2H),7.75(t,J＝7.5Hz,1H),7.62(t,J＝7.8Hz,2H),5.78(dd,J＝17.8,10.4Hz,1H),5.26(s,1H),4.98–4.83(m,5H),4.62(dd,J＝5.6,3.5Hz,2H),4.51(dd,J＝5.4,3.6Hz,2H),4.21(t,J＝6.0Hz,2H),4.08(d,J＝11.6Hz,1H),3.96(d,J＝11.7Hz,1H),2.97–2.84(m,2H),2.67(s,4H),2.63(t,J＝6.0Hz,2H),2.46(d,J＝40.2Hz,8H),2.29–2.24(m,1H),2.15–2.08(m,1H),1.64–1.43(m,6H),0.97(s,3H).13C NMR(100MHz,CDCl3)δ172.2,172.1,158.8,150.7,149.4,147.9,138.3,135.7,129.8,128.8,116.2,111.2,111.0,110.5,69.0,63.5,62.4,61.5,56.7,53.7,53.2,51.0,48.0,42.4,39.9,34.1,29.1,29.1,27.1,15.9.HRMS(ESI)calcd for C35H48ClN4O9S735.2825[M+H]+,found 735.2831.
实施例22
4-(2-((4-(2-(4-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌嗪-1-基)乙氧基)-4-氧代丁酰基)氧基)乙氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例2，区别在于，将(一)中的N-甲基-2-羟基乙胺替换成N-羟乙基哌嗪。
本实施例中制得黄色蜡状液体，产率56％。
1H NMR(400MHz,CDCl3)δ8.06(d,J＝7.3Hz,2H),7.76(t,J＝7.5Hz,1H),7.62(t,J＝7.9Hz,2H),5.78(dd,J＝17.9,10.4Hz,1H),5.26(s,1H),4.99–4.86(m,5H),4.65–4.61(m,2H),4.53–4.47(m,2H),4.21(t,J＝5.9Hz,2H),4.09(d,J＝11.1Hz,1H),3.97(d,J＝11.7Hz,1H),2.92(s,2H),2.62(t,J＝5.9Hz,2H),2.59–2.36(m,12H),2.30–2.24(m,1H),2.16–2.09(m,1H),2.01–1.95(m,2H),1.81(s,2H),1.64–1.41(m,6H),0.98(s,3H).13C NMR(100MHz,CDCl3)δ172.9,172.7,158.8,150.7,149.4,147.9,138.3,135.8,129.8,128.8,116.2,111.3,111.0,110.6,69.1,63.5,62.0,61.3,56.8,53.6,53.2,51.1,48.0,42.4,40.0,34.1,33.3,33.2,29.8,27.2,20.1,15.9.HRMS(ESI)calcd forC36H50ClN4O9S749.2982[M+H]+,found 749.2993.
实施例23
4-((4-((4-(2-(4-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌嗪-1-基)乙氧基)-4-氧代丁酰基)氧基)丁-2-炔-1-基)氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例3，区别在于，将(一)中的N-甲基-2-羟基乙胺替换成N-羟乙基哌嗪。
本实施例中制得黄色蜡状液体，产率60％。
1H NMR(400MHz,CDCl3)δ8.07(d,J＝7.5Hz,2H),7.77(t,J＝7.5Hz,1H),7.63(t,J＝7.8Hz,2H),5.78(dd,J＝17.8,10.4Hz,1H),5.26(s,1H),5.10(s,2H),4.98–4.86(m,5H),4.76(s,2H),4.23(t,J＝5.9Hz,2H),4.09(d,J＝11.6Hz,1H),3.97(d,J＝11.7Hz,1H),2.92(s,2H),2.69–2.62(m,6H),2.48(d,J＝40.6Hz,7H),2.30–2.24(m,1H),2.16–2.08(m,1H),1.89(s,1H),1.64–1.43(m,6H),0.98(s,3H).13C NMR(100MHz,CDCl3)δ172.1,171.6,158.1,150.7,149.4,147.9,138.1,135.8,129.8,128.8,116.2,111.3,111.1,110.7,84.0,78.8,63.5,62.4,58.7,56.7,53.6,53.2,52.2,51.1,48.0,42.4,40.0,34.1,29.1,29.0,27.2,15.9.HRMS(ESI)calcd for C37H48ClN4O9S 759.2825[M+H]+,found759.2811.
实施例24
4-((4-((5-(2-(4-(2-((1R,3R,4S)-3-(3-氯丙-1-烯-2-基)-4-甲基-4-乙烯基环己基)烯丙基)哌嗪-1-基)乙氧基)-5-氧戊二酰基)氧基)丁-2-炔-1-基)氧基)-3-(苯磺酰基)-1,2,5-噁二唑2-氧化物的制备方法基本同实施例4，区别在于，将(一)中的N-甲基-2-羟基乙胺替换成N-羟乙基哌嗪。
本实施例中制得黄色蜡状液体，产率58％
1H NMR(400MHz,CDCl3)δ8.06(d,J＝7.5Hz,2H),7.76(t,J＝7.5Hz,1H),7.63(t,J＝7.9Hz,2H),5.77(dd,J＝17.5,10.7Hz,1H),5.25(s,1H),5.09(s,2H),4.96–4.86(m,5H),4.73(s,2H),4.20(t,J＝5.9Hz,2H),4.08(d,J＝11.5Hz,1H),3.96(d,J＝11.7Hz,1H),2.91(s,2H),2.62(t,J＝5.9Hz,2H),2.42(dt,J＝20.1,7.3Hz,12H),2.29–2.24(m,1H),2.13–2.07(m,1H),1.98–1.91(m,2H),1.63–1.41(m,6H),0.96(s,3H).13C NMR(100MHz,CDCl3)δ172.8,172.1,157.9,150.5,149.3,147.7,137.8,135.8,129.8,128.7,116.2,111.2,111.0,110.6,84.0,78.6,63.3,61.8,58.6,56.7,53.5,53.0,51.9,51.1,47.6,42.1,39.8,39.7,33.9,33.1,32.9,27.0,19.9,15.7.HRMS(ESI)calcd for C38H50ClN4O9S773.2982[M+H]+,found 773.2974.
药效学实验
1.一氧化氮体外释放试验
1.1实验设备与试剂
仪器
多功能酶标仪(美国MD Spectramac M3)
超净工作台(苏州净化)
试剂
磷酸二氢钾(阿拉丁化学试剂有限公司)
磷酸氢二钾(阿拉丁化学试剂有限公司)
L-半胱氨酸(阿拉丁化学试剂有限公司)
1.2实验方法
(1)配制不同浓度的亚硝酸钠溶液绘制校正曲线。
(2)对样品进行处理，配制100μM的溶液，取配好的化合物溶液2.5mL和2.5mL的半胱氨酸溶液37℃孵育120min，不同时间点取样。
(3)设置空白孔和标准孔，收集样本160μL和显色剂0.08mL混匀，37℃静置15min。λ＝550nm，酶标仪测定吸光度OD值，将数值代入标准曲线，即求得NO释放量。
1.3试验结果
表1.实施例1-24制得得β-榄香烯衍生物的NO体外释放结果
由表1中数据可以看出，所有实施例化合物均能有效释放一氧化氮，且绝大多数实施例，随着时间的增加，一氧化氮的释放水平增加。
2.体外抗肿瘤活性评价试验
2.1实验设备与试剂
仪器：
超净工作台(苏州净化设备有限公司)
CO2培养箱(日本SANYO)
倒置生物显微镜(日本OLYMPUS)
酶标仪(美国BioTek)
试剂：
青霉素、链霉素混合液(江苏凯基生物技术股份有限公司)
胰蛋白酶消化液(江苏凯基生物技术股份有限公司)
PBS(江苏凯基生物技术股份有限公司)
Fetal Bovine Serum(GIBCO)
RPMI-1640(江苏凯基生物技术股份有限公司)
DMEM(江苏凯基生物技术股份有限公司)
L-15(江苏凯基生物技术股份有限公司)
CCK8(江苏凯基生物技术股份有限公司)
DMSO(SIGMA)
2.2实验方法
(1)将细胞消化、计数，配制细胞悬液(H520,SW620 5.0×104个/mL，U87MG3.5×104个/mL)，96孔细胞培养板中每孔加入100μL细胞悬液；
(2)96孔细胞培养板置于37℃，5％，CO2培养箱中培养24小时；
(3)用培养基稀释药物至所需工作液浓度，每孔加入100μL相应的含药培养基，同时设立阴性对照组，阳性对照组；
(4)96孔细胞培养板置于37℃，5％，CO2培养箱中培养72小时；
(5)将96孔板进行CCK-8染色，λ＝450nm，测定OD值；
1)每孔加入10μL CCK-8，在培养箱继续培养2-3小时；
2)摇床10分钟轻轻地混匀，去除96孔板中气泡；
3)λ＝450nm，酶标仪读出每孔的OD值，计算抑制率。
(6)利用下列公式(1)，求得肿瘤细胞生长抑制率；所求结果代入IC50计算软件SPSS17.0，求出IC50值，其IC50结果如表1所示。
式(1)
2.3实验结果
表2.实施例1-24对3种人类癌细胞株抗增殖活性的抑制率(1μM)
由表2可知，在1μM浓度下，24个化合物对三种人源肿瘤细胞SW620、U87MG和NCI-H520，均表现出不同程度的抑制作用，且抗增殖活性均显著强于β-榄香烯和13,14-双氯-β-榄香烯，表明NO供体的引入，成功提高β-榄香烯的抗肿瘤作用。
表3.优选实施例化合物对3种人类癌细胞株抗增殖活性的IC50(μM)
如表3所示，绝大部分化合物的抗增殖活性均显著强于β-榄香烯和13,14-双氯-β-榄香烯。对于SW620细胞系，实施例4、14的活性强于β-榄香烯110倍以上；对于U87MG细胞系，实施例2、4、5、6、9、10和22的抗增殖活性强于β-榄香烯250倍以上；对于NCI-H520细胞系，实施例2、4、10、14和22的活性强于β-榄香烯100倍以上。
3.抗脑恶性胶质瘤的体内抗肿瘤活性
3.1受试动物与实验设备
受试动物：
来源、种系、品系：BABLc/裸鼠，上海斯莱克实验动物有限责任公司提供。
实验动物生产许可证：SCXK(沪)2017-0005
合格证编号：20170005040021
实验动物使用许可证：SYXK(苏)2017-0040
日龄：4-5周
性别：雌性
动物数：每组5只，共15只。
实验仪器：
小动物活体三维成像***
Spectrum，PerkinElmer
3.2模型的制备
收集培养的人脑胶质瘤U87MG-LUC细胞悬液，浓度为5X107个/mL，以每只20μL脑原位接种。10％水合氯醛腹腔注射麻醉小鼠，立体定位仪固定老鼠头部。头部皮肤用酒精棉签消毒，沿脑部中线稍偏右切开头部皮肤。取右侧半球，以前囟为基准点，中线旁右2mm，前1mm为进针点，用微型磨钻磨开颅骨。细胞用枪头充分重悬，然后注射器抽取细胞悬液，立体定位仪固定垂直进针3.5mm，退出0.5mm，然后停1min，缓慢注入细胞。注射完后停留1min，缓慢拔针。酒精棉签消毒皮肤，用针线缝合皮肤后将小鼠放回饲养笼自然苏醒。
3.3分组与给药
细胞接种14天后，将动物随机分组，每组5只。同时，各组裸鼠开始给药，给药方案见组别及给药方案。实验结束后，随即处死裸鼠，手术剥取脑组织，拍照、称重。
3.4活体成像检测
每组选2只动物分别在给药前，给药1周，2周，3周后腹腔注射Luc荧光底物,100μL/只，10min后小鼠异氟烷麻醉，放置于仪器暗箱载物台上，进行活体成像检测，观察小鼠脑部原位瘤的荧光情况。
3.5实验结果
如图6和图7所示，β-榄香烯和实施例4给药3周后，小鼠恶性脑胶质瘤的生长得到有效抑制。模型组肿瘤体积持续增大(图6和图7A)，而β-榄香烯和实施例4治疗后，肿瘤体积持续受到抑制，明显小于模型组。从脑重来看，β-榄香烯和实施例4组的重量明显轻于模型组(图7B)，小鼠的生活和身体状况也明显优于模型组。另一方面，模型组胶质瘤的荧光强度持续快速增加，而β-榄香烯和实施例4组的荧光强度则急剧下降。在第一周，实施例4的抑制活性超过80％，明显高于β-榄香烯(>20％)。在第二周，β-榄香烯和实施例4均达到>80％的抑制。在给药的最后一周，实施例4阻止肿瘤生长>90％，表现出强效的抗恶性脑胶质瘤活性。总之，β-榄香烯和实施例4均显示出有效的治疗活性，但实施例4显示出比β-榄香烯更强的抑制作用。
以上结果表明NO供体的引入有效增强了β-榄香烯在体内的抗肿瘤作用，是基于β-榄香烯抗肿瘤药物研发的可行策略。
此外应理解，在阅读了本发明的上述描述内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。
Claims (7)
1.14-氯-β-榄香烯一氧化氮供体型衍生物，或其可药用盐，或其对映异构体、非对映异构体，其特征在于，所述14-氯-β-榄香烯一氧化氮供体型衍生物具有如下通式(I)：
式(I)中：
R1为含氮和氧原子的环状醇胺结构，选自
R2选自C1-10链烷基；
R3选自C1-10链烷基或C2-10炔基。
2.根据权利要求1所述的14-氯-β-榄香烯一氧化氮供体型衍生物，或其可药用盐，或其对映异构体、非对映异构体，其特征在于，式(I)中：R2选自C2-6的链烷基，R3选自C2-6的链烷基或C2-6的炔基。
3.根据权利要求1所述的14-氯-β-榄香烯一氧化氮供体型衍生物，或其可药用盐，或其对映异构体、非对映异构体，其特征在于，式(I)中：R2选自-CH2CH2-、-CH2CH2CH2-、-CH2CH2CH2CH2-、-CH2CH2CH2CH2CH2-，R3选自-CH2CH2-、-CH2CH2CH2-、-CH2CH2CH2CH2-、-CH2CH2CH2CH2CH2-、-CH2C≡C-、-C≡CCH2-、-CH2C≡CCH2-、-CH2CH2C≡CCH2-、-CH2CH2C≡CCH2CH2-、-CH2C≡CCH2CH2-。
4.根据权利要求1所述的14-氯-β-榄香烯一氧化氮供体型衍生物，或其可药用盐，或其对映异构体、非对映异构体，其特征在于，所述14-氯-β-榄香烯一氧化氮供体型衍生物选自如下通式(I-3)～(I-6)：
式(I-3)～(I-6)中，R2选自C2-6的链烷基，R3选自C2-6的链烷基或C2-6的炔基。
5.根据权利要求1所述的14-氯-β-榄香烯一氧化氮供体型衍生物，或其可药用盐，或其对映异构体、非对映异构体，其特征在于，所述14-氯-β-榄香烯一氧化氮供体型衍生物选自如下结构所示的化合物9～24：
6.根据权利要求1～5任一权利要求所述的14-氯-β-榄香烯一氧化氮供体型衍生物，或其可药用盐，或其对映异构体、非对映异构体在制备抗肿瘤药物中的应用。
7.根据权利要求6所述的应用，其特征在于，所述肿瘤包括肺癌、结肠癌、恶性脑胶质瘤。
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