双-(10-去氧双氢青蒿素)-间苯三酚衍生物的制备及应用的制作方法

文档序号:29356272发布日期:2022-03-23 00:04阅读:325来源:国知局
双-(10-去氧双氢青蒿素)-间苯三酚衍生物的制备及应用的制作方法

1.本发明属于生物医药领域,涉及一种间苯三酚衍生物,具体来说是一种双-(10-去氧双氢青蒿素)-间苯三酚衍生物的制备及应用。


背景技术:

2.青蒿素是上世纪70年代由中国科学家从菊科植物黄花蒿(artemisia annua.l.)中提取发现的抗疟疾药物,它是一类具有特殊过氧桥结构的倍半萜内酯类化合物,也是由中国发现的第一个被国际公认的天然药物。但青蒿素口服不易吸收,生物利用度低,难以制成合适的剂型。科学家们通过对青蒿素结构改造,合成了抗疟效果更好的蒿甲醚、青蒿琥酯等青蒿素衍生物。除了良好的抗疟活性,近年来大量研究表明一些青蒿素衍生物不但对多种肿瘤细胞有杀伤作用,而且具有良好的免疫抑制作用,对系统性红斑狼疮,类风湿性关节炎,过敏性皮炎等自身免疫性疾病均有较好的治疗效果。
3.在专利cn110343121b中,报道了双-(10-去氧双氢青蒿素)-间苯三酚衍生物1-13:
4.[0005][0006]
尽管这些化合物显示出了良好的抗肿瘤活性和免疫抑制活性,然而在动物体内实验中这些化合物显示出较高的毒性,特别在高剂量下动物耐受性不佳,甚至会导致动物死
亡,因此这类化合物的治疗窗口尚需扩大,才能具有实用价值。为了降低这类青蒿素二聚体的毒性,我们首先合成了化合物14-18:
[0007][0008]
但药理实验结果显示,这几个化合物依然显示出很高的毒性,治疗窗口非常窄小。
[0009]
为了进一步降低这类青蒿素二聚体的毒性,我们采用了两种结构修饰的策略:一是在双-(10-去氧双氢青蒿素)-间苯三酚的酚羟基上引入大位阻的酯或醚来改变药物分子体内动力学特性;第二种策略是在苯环上引入可以成盐的氨基、氨基酸。这样不仅可以增加位阻以改变药物分子在体内的动力学特征,而且可以增加水溶性,以改变药物的脂水分布系数,进而降低毒性。我们采用cck-8法测试这些新的双-(10-去氧双氢青蒿素)-间苯三酚衍生物对vero细胞的毒性。vero细胞系是从非洲绿猴的肾脏上皮细胞中分离培养出来的。这个细胞系由日本千叶大学的yasumura和kawakita于1962年3月27日扩增出来。该细胞系取“verda reno”(世界语意为

绿色的肾脏’)的简写而命名为"vero"。测试化合物对vero细胞的体外增值抑制活性,可以反映出化合物的细胞毒性,试验测定的ic
50
值越大,说明化合物的细胞毒性就越低。


技术实现要素:

[0010]
针对现有技术中的问题,本发明提供了一种双-(10-去氧双氢青蒿素)-间苯三酚衍生物及其制备方法和用途,所述的这种双-(10-去氧双氢青蒿素)-间苯三酚衍生物及其制备方法和用途要解决现有技术中的双-(10-去氧双氢青蒿素)-间苯三酚衍生物具有的高毒性、治疗窗口窄小的技术问题。
[0011]
本发明提供了一种双-(10-去氧双氢青蒿素)-间苯三酚衍生物,其结构式如下所示:
[0012][0013]
通式(i)中,两分子10-去氧双氢青蒿素的10-位碳通过碳碳键与苯环相连;10-去氧双氢青蒿素10-位碳的构型可以是r或s构型。
[0014]
通式(i)中,r1、r2、r3可以相同也可以不相同。
[0015]
通式(i)中,r1,r2,r3各自分别包括h,-oh,-xr5,
[0016]
其中xr5包括醚(-or5)、羧酸酯(r5coo-)、碳酸酯(-ocoor5)、磺酸酯(-oso2r5)、亚磺酸酯(-osor5)、磷酸酯(-opo3r5)、亚磷酸酯(-opo2r5)、氨基甲酸酯(-oconhr5)、异脲(-ocnhnhr5)中的任意一种;
[0017]
r5为1-15个碳的烷基、烯基,炔基或其衍生物;1-15个碳并含1-6个卤素的卤代烷基、卤代烯基或卤代炔基或其衍生物;6-20个碳原子的芳基、卤代芳基、酚或多元酚或其衍生物;6-20个碳原子的芳香羧酸或其衍生物;1-15个碳原子的醇或多元醇或其衍生物;1-15个碳原子的羧酸,多元羧酸或其衍生物;1-15个碳原子的磺酸或其盐;1-15个碳原子的胺、胺盐、季胺盐;1-15个碳原子的酰胺或其衍生物;1-15个碳原子的卤代烷氧基;2-15个碳原子的烷氧基醇、烷氧基多元醇以及它们的醚、羧酸酯、磺酸酯或硫酸酯或其盐;2-15个碳原子的烷氧基羧酸,烷氧基多元羧酸,烷氧基氨基酸或其盐;2-15个碳原子的烷氧基磺酸或其盐;2-15个碳原子的烷氧基胺或其胺盐和季胺盐;2-15个碳原子的烷氧基酰胺;含3-15个碳原子的烷氧基烯烃;含3-15个碳原子的烷氧基炔烃;含3-20个碳原子的氨基酸或其衍生物;含3-20个碳原子的氨基醇或其衍生物;4-15个碳的环烷基醚;含有1-4个杂原子的五元-八元的杂环或并杂环或其衍生物;
[0018]
r4包括-h,-oh,卤素,硝基,氨基,磺酸基,-r5或-xr5;
[0019]
其中xr5包括醚(-or5)、羧酸酯(r5coo-)、碳酸酯(-ocoor5)、磺酸酯(-oso2r5)、亚磺酸酯(-osor5)、磷酸酯(-opo3r5)、亚磷酸酯(-opo2r5)、氨基甲酸酯(-oconhr5)、异脲(-ocnhnhr5)中的任意一种;
[0020]
r5为下述的任意一个基团:
[0021]
1-15个碳的烷基、烯基、炔基或其衍生物;1-15个碳并含1-6个卤素的卤代烷基、卤代烯基或卤代炔基或其衍生物;6-20个碳原子的芳基、卤代芳基、酚或多元酚或其衍生物;
6-20个碳原子的芳香羧酸或其衍生物;1-15个碳原子的醇或多元醇或其衍生物;1-15个碳原子的羧酸,多元羧酸或其衍生物;1-15个碳原子的磺酸或其盐;1-15个碳原子的胺、胺盐、季胺盐;1-15个碳原子的酰胺或其衍生物;1-15个碳原子的卤代烷氧基;2-15个碳原子的烷氧基醇、烷氧基多元醇以及它们的醚、羧酸酯、磺酸酯或硫酸酯或其盐;2-15个碳原子的烷氧基羧酸,烷氧基多元羧酸,烷氧基氨基酸或其盐;2-15个碳原子的烷氧基磺酸或其盐;2-15个碳原子的烷氧基胺或其胺盐和季胺盐;2-15个碳原子的烷氧基酰胺;含3-15个碳原子的烷氧基烯烃;含3-15个碳原子的烷氧基炔烃;含3-20个碳原子的氨基酸或其衍生物;含3-20个碳原子的氨基醇或其衍生物;4-15个碳的环烷基醚;含有1-4个杂原子的五元-八元的杂环或并杂环或其衍生物。
[0022]
通式(i)中,r4可以与r2、r3形成环状结构。
[0023]
本发明通式(i)中,双-(10-去氧双氢青蒿素)-间苯三酚衍生物的异构体包括其所有的异构体,如位置异构体,立体异构体和光学异构体。
[0024]
本发明所述的一种双-(10-去氧双氢青蒿素)-间苯三酚衍生物,当r4为-chxnr6r7时,可以用下列方法进行制备:
[0025][0026]
该制备方法的特征在于:在醛或醛的类似物存在下,胺nhr6r7和双-(10-去氧双氢青蒿素)-间苯三酚及其衍生物发生苯环上的胺甲基化反应。
[0027]
其中,r1,r2,r3的定义范围和通式(i)相同;r6和r7可以相同也可以不相同,r6和r7也可以形成环状结构;
[0028]
其中,x,r6,r7分别为下述的任意一个基团:
[0029]-h,1-15个碳的烷基、烯基,炔基或其衍生物;1-15个碳并含1-6个卤素的卤代烷基、卤代烯基或卤代炔基或其衍生物;6-20个碳原子的芳基、卤代芳基、酚或多元酚或其衍生物;6-20个碳原子的芳香羧酸或其衍生物;1-15个碳原子的醇或多元醇或其衍生物;1-15个碳原子的羧酸,多元羧酸或其衍生物;1-15个碳原子的磺酸或其盐;1-15个碳原子的胺、胺盐、季胺盐;1-15个碳原子的酰胺或其衍生物;1-15个碳原子的卤代烷氧基;2-15个碳原子的烷氧基醇、烷氧基多元醇以及它们的醚、羧酸酯、磺酸酯或硫酸酯或其盐;2-15个碳原子的烷氧基羧酸,烷氧基多元羧酸,烷氧基氨基酸或其盐;2-15个碳原子的烷氧基磺酸或其盐;2-15个碳原子的烷氧基胺或其胺盐和季胺盐;2-15个碳原子的烷氧基酰胺;含3-15个碳原子的烷氧基烯烃;含3-15个碳原子的烷氧基炔烃;含3-20个碳原子的氨基酸或其衍生物;含3-20个碳原子的氨基醇或其衍生物;4-15个碳的环烷基醚;含有1-4个杂原子的五元-八元的杂环或并杂环或其衍生物。
[0030]
该方法的特征在于反应中使用的醛包括各种脂肪醛和芳香醛,如甲醛,或三聚甲醛,或多聚甲醛,乙醛,丙醛,苯甲醛等。醛的类似物包括:甲醛类似物如二氯甲烷,二溴甲烷;其他脂肪醛和芳香醛类似物如相应的缩醛、二氯或二溴等价物。
[0031]
该制备方法中的胺包括nh3,甲胺,乙胺,二甲胺,二乙胺,二丙胺,二丁胺,甲乙胺,乙醇胺,二乙醇胺,氮杂环丙氨,氮杂环丁胺,吡咯,哌啶,吗啡啉,哌嗪,n-甲基哌嗪等。
[0032]
该方法的制备方法,所述反应所使用的溶剂为质子溶剂、非质子溶剂或质子溶剂和非质子溶剂的混合溶剂,质子溶剂包括水,甲醇,乙醇,正丙醇,异丙醇,正丁醇,叔丁醇,甲酸,乙酸,丙酸,丁酸等;非质子溶剂包括乙醚,二氯甲烷,1,2-二氯乙烷,四氢呋喃,1,4-二氧六环,乙腈,乙二醇二甲醚,甲基叔丁基醚等;所述反应温度在摄氏温度-78~140℃。
[0033]
上述反应的粗产物可以用溶剂提取法,沉淀法,结晶法进一步纯化,也可以用柱层析法进行纯化,填料用硅胶,碳18,mci凝胶,大孔树脂或氧化铝,洗脱剂可以用石油醚-丙酮,石油醚-乙酸乙酯,石油醚-二氯甲烷,二氯甲烷-甲醇,乙腈-水不同比例的混合。
[0034]
本发明还提供了上述具有通式(i)的双-(10-去氧双氢青蒿素)-间苯三酚衍生物的可药用盐或其前药分子。
[0035]
优选的,所述通式(i)双-(10-去氧双氢青蒿素)-间苯三酚衍生物包括下述具体化合物19-67:
[0036]
[0037]
[0038]
[0039]
[0040]
[0041][0042]
实验证明,化合物19~67的细胞毒性与原型化合物1相比,其对vero细胞的毒性显著下降(见实施例55),而这些化合物在pha刺激的人pbmc体外增殖抑制实验中体现出与化合物1类似的抑制活性,使得其治疗窗口得到了极大的改善(实施例56)。
[0043]
在上述化合物19~67中,我们挑选了毒性较小的化合物26,34,42,59和67进行了动物体内实验,我们首先采用了dnfb(2,4-二硝基氟苯)诱导迟发性超敏反应的耳肿胀模型,以化合物1做对照,评估了化合物26、34、42、59和67对耳肿胀的抑制效果(实施例57)。实验发现,化合物1在高剂量下(8mpk),小鼠全部死亡,但在中低剂量下,对dnfb(2,4-二硝基氟苯)诱导迟发性超敏反应的小鼠耳肿胀具有显著抑制作用。而化合物26,34,42,59和67在低中高三个剂量组不仅对小鼠耳肿胀具有显著抑制作用,而且呈现显著的量效关系。并且在高剂量组中,小鼠的体重没有下降。因此,我们对这五个化合物提高了剂量,并采用绵羊红细胞诱导的迟发性超敏反应的跖肿胀模型对这些化合物的活性和治疗窗口进行了验证(实施例58)。实验证明,上述化合物在剂量增加到18mpk时,仍然对小鼠的体重没有影响,且在低中高三个剂量组中,显著抑制绵羊红细胞诱导的小鼠跖肿胀,且上述抑制作用呈显著的剂量相关性。
[0044]
在本发明中,双-(10-去氧双氢青蒿素)-间苯三酚衍生物的可药用盐,包括锂盐,钾盐,钠盐,钙盐,镁盐,或者与天然的和非天然的有机含氮化合物形成的有机盐;与盐酸,硫酸,磷酸等无机酸生成的盐;与苹果酸,马来酸,甲磺酸等有机酸生成的盐。
[0045]
在本发明中,双-(10-去氧双氢青蒿素)-间苯三酚衍生物的前药分子,是指药物经化学结构修饰后得到的在体内能迅速转化为上述通式(i)所示母体化合物的化合物,设计前药分子的目的在于增加药物的生物利用度,加强靶向性,降低药物的毒性和副作用等。
[0046]
在本发明的另一方面,双-(10-去氧双氢青蒿素)-间苯三酚衍生物还包括其与其他药物的组合物,其含有治疗有效量的上述双-(10-去氧双氢青蒿素)-间苯三酚衍生物或其可药用盐、或其前药分子,以及一种或多种药学上可接受的载体、稀释剂或赋形剂。
[0047]
上述可接受的载体是无毒的、能辅助施用并且对结合物的治疗效果没有不利影响。此类载体可以是本领域的技术人员通常能得到的任何固体赋形剂、液体赋形剂、半固体赋形剂或者在气雾剂组合物中的气体赋形剂。固体药物赋形剂包括淀粉、纤维素、滑石、葡萄糖、乳糖、蔗糖、明胶、麦芽、稻米、面粉、白垩、硅胶、硬脂酸镁、硬脂酸钠、甘油硬脂酰酯、氯化钠、无水脱脂乳等。液体和半固体赋形剂可以选自甘油、丙二醇、水、乙醇和各种油,包括那些源于石油、动物、植物或人工合成的油,例如,花生油、豆油、矿物油、芝麻油等,优选的液体载体,特别是用于可注射溶液的,包括水、盐水、葡萄糖水溶液和甘醇。另外还可以在组合物中加入其它辅剂如香味剂、甜味剂等。
[0048]
本发明的药物或药物组合物可以通过口服或其他给药方式给药,如注射,透皮给药,喷雾给药,直肠给药,阴道给药等。优选的给药方式是注射或口服,它可根据疾病程度调节。
[0049]
本发明的药物组合物可以和其他治疗自身免疫性疾病药物联合用药,所述的其他治疗自身免疫性疾病药物包括以下几种:
[0050]
(1)抗代谢类:硫唑嘌呤(aza)、氨甲喋呤、霉酚酸酯(mmf)等;
[0051]
(2)烃化剂:环磷酰胺等;
[0052]
(3)皮质固醇类:强的松、地塞米松等;
[0053]
(4)抗生素类:csa、fk506、雷帕霉素等;
[0054]
(5)抗体类:抗淋巴细胞球蛋白(alg)、单克隆t淋巴细胞抗体(okt3)等;
[0055]
(6)中草药:雷公藤多苷、冬虫夏草制剂等。
[0056]
本发明药物组合物的各种剂型可以按照药学领域的常规方法制备。例如使该化合物与一种或者多种载体混合,然后将其制成所需的剂型,如片剂、药丸、胶囊、半固体、粉末、缓释剂型、溶液、混悬液、配剂、气雾剂等。
[0057]
在本发明的另一方面,还提供了上述双-(10-去氧双氢青蒿素)-间苯三酚衍生物或其可药用盐、或其前药分子在制备治疗自身免疫性疾病相关病症的药物中的应用。
[0058]
上述自身免疫性疾病包括红斑狼疮、类风湿关节炎、强直性脊柱炎、i型糖尿病、肾病综合征、干燥综合征、多发性硬化症、银屑病、皮肌炎、过敏性鼻炎、顽固性荨麻疹、自身免疫性溶血性贫血、特发性血小板减少性紫癜、克罗恩病、非特异性溃疡性结肠炎、硬皮病、人体器官移植后的排斥反应等。
[0059]
本发明的双-(10-去氧双氢青蒿素)-间苯三酚衍生物,合成简单,体外vero细胞增值抑制试验证明,这些化合物不但细胞毒性小,进一步的体内外药效试验也证明这些化合
物具有显著的免疫抑制活性,作为免疫抑制药物有广阔的应用前景。
附图说明
[0060]
图1青蒿素二聚体对迟发性超敏反应的耳肿胀的抑制作用,显示第七天右耳厚度减去第六天右耳厚度的厚度差。
[0061]
图2青蒿素二聚体对迟发性超敏反应的跖肿胀的抑制作用,显示第6天左右足跖的厚度差。
具体实施方式
[0062]
实施例1化合物14的制备
[0063][0064]
在氮气保护下,将双氢青蒿素(1.30g,4.6mmol)溶于20ml干燥二氯甲烷中,将反应体系降温至-30℃,缓慢滴加dast(二乙胺基三氟化硫,0.66ml,4.6mmol),然后在-30℃下继续搅拌1小时,tlc监控反应,直至原料消耗完毕,得到中间体14-1。然后直接向反应液中加入1,3,5-三甲氧基苯(1.55g,9.2mmol)并降温至-78℃后,缓慢滴加bf3·
et2o(692μl,5.5mmol),然后反应温度由-78℃逐渐升高到室温。tlc监控至反应结束,用饱和nahco3水溶液(20ml)淬灭反应,分离有机相,水相用10ml二氯甲烷萃取两次。合并有机相,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=6/1),得1.868g白色固体中间体14-2,收率93.5%。
[0065]1h nmr(400mhz,dmso-d6)δ6.23(dd,j=8.5,2.2hz,2h),5.29(s,1h),4.92(d,j=10.9hz,1h),3.88-3.63(m,9h),3.28

3.14(m,1h),2.18(dd,j=13.8,3.7hz,1h),1.99(s,1h),1.86

1.74(m,1h),1.72

1.51(m,3h),1.47(d,j=5.5hz,3h),1.20(d,j=31.1hz,4h),0.91(d,j=6.3hz,4h),0.46(d,j=7.3hz,3h).
[0066]
在氮气保护下,将双氢青蒿素(0.659g,2.3mmol)溶于15ml干燥的二氯甲烷中,降温至-30℃,缓慢滴加dast(300μl,2.3mmol),在-30℃下继续搅拌至原料消耗完毕,得到含14-1的反应液。向该反应液中加入中间体14-2(1.50g,3.5mmol)并降温至-78℃后,缓慢滴加bf3·
et2o(435μl,3.5mmol),滴毕,反应液自然升温,tlc监控反应至反应结束。用饱和nahco3水溶液(10ml)淬灭反应,分离有机相,水相用10ml二氯甲烷萃取两次。合并有机相,用无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=6/
1),得60mg白色固体产物14,收率3.7%。
[0067]1h nmr(400mhz,dmso-d6)δ6.49(s,1h),5.36(d,j=3.5hz,2h),4.99(d,j=10.8hz,1h),4.78(d,j=10.7hz,1h),3.96-3.68(m,9h),3.31-3.15(m,2h),2.28-2.14(m,2h),2.00(d,j=14.1hz,2h),1.81(s,2h),1.68(d,j=13.6hz,2h),1.60(s,3h),1.52-1.46(m,3h),1.41(s,3h),1.23(dd,j=20.2,8.3hz,8h),1.03-0.81(m,10h),0.39(dd,j=14.5,7.2hz,5h).
[0068]
13
c nmr(101mhz,dmso-d6)δ161.06,160.94,158.73,115.59,114.28,113.92,103.58,103.57,95.54,92.09,92.05,81.01,80.87,69.91,69.60,68.27,63.68,60.23,57.68,57.38,56.56,52.40,46.20,46.10,36.86,36.77,36.59,34.26,29.14,28.77,26.20,26.02,24.86,20.99,20.75,20.67,14.56,13.86,13.77.
[0069]
ms(esi):[m+na]
+
=723.4
[0070]
实施例2化合物15的制备
[0071][0072]
氮气保护下,将化合物1(100mg,0.15mmol)溶解在15ml丙酮中,加入3-溴丙烯(66μl,0.75mmol)和碳酸钾固体(62mg,0.45mmol),将反应体系升温至70℃,回流6小时。tlc监控反应至反应结束后,反应液用30ml乙酸乙酯稀释,然后用饱和食盐水洗涤两次,有机相用无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=5/1),得78mg白色固体产物15,收率74.7%。
[0073]1h nmr(400mhz,dmso-d6)δ8.54(d,j=3.4hz,2h),6.02(q,j=10.9hz,2h),5.66(s,2h),5.41(dd,j=17.3,1.8hz,1h),5.26(dd,j=10.7,1.7hz,1h),5.15(d,j=11.2hz,1h),5.08(d,j=11.2hz,1h),4.47(dd,j=12.3,4.6hz,2h),2.65-2.55(m,2h),2.25(d,j=13.5hz,2h),2.05(d,j=14.7hz,2h),1.92-1.78(m,2h),1.67(d,j=12.0hz,5h),1.55(d,j=13.0hz,4h),1.34(d,j=6.5hz,7h),1.25(d,j=9.3hz,4h),1.01(d,j=9.5hz,2h),0.92(d,j=6.2hz,6h),0.62(d,j=7.1hz,3h),0.56(d,j=7.1hz,3h).
[0074]
13
c nmr(101mhz,dmso-d6)δ156.87,155.86,154.23,134.19,116.72,106.63,104.87,104.73,104.69,93.75,91.61,91.39,81.70,81.60,70.99,68.52,51.52,45.60,36.70,35.99,34.22,31.64,31.18,26.22,26.09,24.92,20.79,20.52,13.63,13.53.
[0075]
ms(esi):[m+na]
+
=721.4
[0076]
实施例3化合物16的制备
[0077][0078]
将化合物2(190mg,0.26mmol)溶解在20ml甲醇、20ml四氢呋喃、2ml水的混合溶液中并用冰水浴冷到5℃,然后加入lioh固体(61mg,2.6mmol),并在5℃下搅拌1小时。tlc监控反应结束后,滴加0.05m的稀盐酸至反应液ph为5左右,室温减压浓缩除去大部分溶剂,残余物用20ml水稀释,乙酸乙酯萃取两次。合并有机相,用无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=15/1),得113mg白色固体产物16,收率61.8%。
[0079]1h nmr(400mhz,dmso-d6)δ8.54(d,j=2.4hz,2h),5.85(s,1h),5.63(d,j=15.9hz,2h),5.15(d,j=11.2hz,1h),5.08(d,j=11.2hz,1h),4.54(d,j=5.1hz,2h),2.58(d,j=7.3hz,2h),2.26(t,j=12.4hz,2h),2.04(d,j=11.1hz,2h),1.85(s,2h),1.71

1.45(m,9h),1.33(d,j=7.4hz,8h),1.25(d,j=2.3hz,3h),1.02(s,2h),0.91(d,j=6.3hz,6h),0.60(dd,j=15.1,7.1hz,6h)..
[0080]
13
c nmr(101mhz,dmso-d6)δ170.50,156.73,155.53,154.29,106.95,104.94,104.75,104.70,93.38,91.63,91.37,81.68,81.59,71.02,65.44,51.52,45.65,36.73,35.99,34.18,31.63,31.14,26.22,26.08,24.91,20.79,20.70,20.52,13.62,13.54.
[0081]
ms(esi):[m+h]
+
=717.3
[0082]
实施例4化合物17的制备
[0083][0084]
化合物2(200mg,0.27mmol)溶解在15ml四氢呋喃和15ml甲醇的混合溶液中,然后滴加氨水(30%,3.0ml,40.2mmol),室温搅拌过夜。tlc监控反应完全后,室温减压蒸干溶剂,残余物用30ml乙酸乙酯溶解。溶液用饱和食盐水洗涤两次,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=2/1~1/2),得96mg白色固体产物17,收率49.7%。
[0085]1h nmr(400mhz,dmso-d6)δ8.57(d,j=8.7hz,2h),7.47(s,1h),7.34(s,1h),5.95(s,1h),5.63(d,j=19.5hz,2h),5.29(d,j=11.2hz,1h),5.09(d,j=11.2hz,1h),4.45(d,j=14.6hz,1h),4.20(d,j=14.6hz,1h),2.58(s,2h),2.27(t,j=12.4hz,2h),2.06(d,j=12.7hz,2h),1.85(s,2h),1.69(s,5h),1.58(s,2h),1.34(d,j=8.1hz,7h),1.30-1.21(m,6h),1.02(d,j=10.4hz,2h),0.92(d,j=5.1hz,6h),0.63(d,j=7.0hz,3h),0.54(d,j=6.9hz,3h).
[0086]
13
c nmr(101mhz,dmso-d6)δ170.14,156.83,155.33,154.32,107.14,105.01,104.76,104.71,104.68,93.84,91.60,91.39,81.71,81.59,70.98,67.49,60.23,51.51,45.65,45.60,37.06,36.75,35.99,34.22,31.61,31.10,30.31,26.23,26.08,24.95,21.24,20.79,20.50,14.57,13.61.
[0087]
ms(esi):[m+na]
+
=738.3
[0088]
例5化合物18的制备
[0089][0090]
在氮气保护下,将化合物1(1.0g,1.51mmol)溶解在30ml二氯甲烷中,加入三乙胺(630μl,4.53mmol),将反应体系冷却至0℃,缓慢滴加含丙烯酰氯(249μl,3.02mmol)的5ml二氯甲烷溶液,然后在0~5℃下继续搅拌。tlc监控反应结束后,加入2ml甲醇淬灭反应,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=5/1~3/1),得96mg白色固体产物18,收率8.3%。
[0091]1h nmr(400mhz,dmso-d6)δ8.87(s,1h),6.56(dd,j=12.2,6.0hz,2h),6.46-6.28(m,3h),6.25-6.15(m,1h),6.00(dd,j=7.9,3.9hz,1h),5.74(s,1h),5.36(s,1h),5.10(d,j=10.9hz,1h),4.96(d,j=11.2hz,1h),2.89(d,j=3.5hz,1h),2.65(s,1h),2.27(d,j=10.4hz,1h),2.11(dd,j=31.1,12.9hz,2h),1.89(dd,j=46.3,22.4hz,3h),1.76-1.58(m,6h),1.45(d,j=55.2hz,8h),1.18(s,6h),1.06-0.81(m,8h),0.66-0.41(m,6h).
[0092]
13
c nmr(101mhz,dmso-d6)δ164.27,163.57,155.11,150.01,146.88,134.47,131.81,129.51,127.86,118.57,114.94,110.16,104.96,103.73,92.15,91.55,81.63,80.76,71.66,67.93,52.32,51.40,45.94,45.29,36.74,36.58,36.43,35.89,34.22,34.06,30.57,30.41,26.19,25.90,24.87,21.01,20.69,20.44,20.18,13.69,13.26.
[0093]
ms(esi):[m+na]
+
=789.3
[0094]
实施例6化合物19的制备
[0095][0096]
氮气保护下,将化合物1(660mg,1.0mmol)溶解在200ml二氯甲烷中,室温下滴加哌啶(8.0ml,87.4mmol),然后室温搅拌1小时。tlc监控至反应结束后,反应液依次用饱和磷酸二氢钠水溶液、饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=5/1~5/3),得471mg淡黄色固体产物19,收率62.3%。
[0097]1h nmr(400mhz,dmso-d6)δ8.58(s,1h),8.31(s,1h),5.63(d,j=11.7hz,2h),5.07
(dd,j=10.9,7.4hz,2h),3.65(s,2h),2.67

2.54(m,2h),2.46(s,2h),2.26(t,j=13.0hz,2h),2.05(d,j=13.8hz,2h),1.85(s,2h),1.67(d,j=11.3hz,6h),1.50(d,j=29.7hz,10h),1.35(dd,j=18.0,9.6hz,9h),1.24(dd,j=17.0,10.3hz,3h),1.00(d,j=11.1hz,2h),0.92(d,j=6.0hz,7h),0.60(dd,j=11.0,7.3hz,6h).
[0098]1h nmr(400mhz,cdcl3+d2o)δ5.43(d,j=5.5hz,2h),5.24(d,j=11.3hz,1h),5.14(d,j=11.2hz,1h),3.79(d,j=14.2hz,1h),3.69(d,j=14.2hz,1h),2.90

2.71(m,3h),2.44(dd,j=17.4,6.9hz,3h),2.23

1.98(m,3h),1.91(dd,j=6.5,3.2hz,2h),1.89

1.73(m,4h),1.61(s,10h),1.46(d,j=2.3hz,7h),1.42

1.22(m,5h),1.22

0.82(m,9h),0.71(dd,j=21.0,7.1hz,6h)..
[0099]
13
c nmr(101mhz,dmso-d6)δ156.17,153.95,152.99,104.67,104.06,103.73,99.91,91.57,91.48,81.68,81.58,71.18,71.02,54.36,53.40,51.56,45.69,36.80,36.71,36.01,34.23,31.69,31.33,26.22,26.20,25.89,24.94,24.01,20.82,20.75,20.54,20.50,13.68,13.41
[0100]
ms(esi):[m+h]
+
=756.5
[0101]
实施例7化合物20的制备
[0102][0103]
氮气保护下,将化合物1(300mg,0.46mmol)溶解在50ml二氯甲烷中,加入哌嗪(100mg,116.0mmol)和多聚甲醛(30mg),室温下搅拌2小时。tlc监控反应结束后,反应液用饱和食盐水洗涤至中性。有机相经无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1~20/1),得235mg白色固体产物20,收率68.2%。
[0104]1h nmr(400mhz,dmso-d6)δ8.58(s,1h),8.29(s,1h),5.63(d,j=15.0hz,2h),5.08(dd,j=11.2,4.3hz,2h),3.81(d,j=1.3hz,2h),2.59(s,5h),2.26(t,j=13.2hz,2h),2.05(d,j=14.3hz,2h),1.94

1.81(m,2h),1.78(s,4h),1.67(d,j=13.5hz,6h),1.55(s,4h),1.32(t,j=6.0hz,9h),1.28

1.21(m,3h),0.99(s,2h),0.92(d,j=6.3hz,6h),0.60(dd,j=10.9,7.2hz,6h).
[0105]
13
c nmr(101mhz,dmso-d6)δ155.63,154.09,153.18,104.67,104.44,103.82,99.68,91.56,91.50,81.67,81.57,71.16,71.00,53.28,52.08,51.56,45.70,36.84,36.72,36.01,34.25,31.66,31.31,26.21,24.95,20.82,20.54,20.49,13.69,13.50.
[0106]
ms(esi):[m+h]
+
=757.4
[0107]
实施例8化合物21的制备
[0108][0109]
氮气保护下,将化合物1(660mg,1.0mmol)溶解在50ml二氯甲烷中,在室温下缓慢滴加n-甲基哌嗪(10.0ml,90.0mmol),然后在室温下搅拌72小时。tlc监控反应结束后,反应液依次用饱和磷酸二氢钠水溶液、饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1~50/1),得638mg淡黄色固体产物21,收率82.7%。
[0110]1h nmr(400mhz,dmso-d6)δ8.61(s,1h),8.33(s,1h),5.63(d,j=14.2hz,2h),5.07(t,j=10.4hz,2h),3.68(s,2h),2.66-2.51(m,6h),2.24(d,j=13.3hz,3h),2.18(s,3h),2.05(d,j=13.6hz,3h),1.85(s,2h),1.67(d,j=11.3hz,6h),1.54(d,j=11.1hz,4h),1.33(d,j=1.8hz,8h),1.30-1.12(m,3h),1.00(d,j=8.7hz,2h),0.92(d,j=5.8hz,6h),0.59(dd,j=14.3,7.1hz,6h).
[0111]
13
c nmr(101mhz,dmso-d6)δ155.83,154.03,153.10,104.67,104.25,103.75,99.72,91.58,91.49,81.67,81.56,71.17,71.00,54.94,53.51,52.17,51.56,45.96,45.69,36.82,36.72,36.01,34.24,31.69,31.32,26.21,24.95,20.82,20.77,20.54,20.50,13.68,13.43.
[0112]
ms(esi):[m+h]
+
=771.4
[0113]
实施例9化合物22的制备
[0114][0115]
氮气保护下,将化合物1(300mg,0.46mmol)溶解在50ml二氯甲烷中,室温下缓慢滴加吗啉(1.0ml,1.1mmol)后,加入多聚甲醛(30mg),室温下搅拌2小时。tlc监控反应结束后,反应液用饱和食盐水洗涤至中性,经无水硫酸钠干燥,减压浓缩。残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1~20/1),得246mg白色固体产物22,收率71.3%。
[0116]1h nmr(400mhz,dmso-d6)δ8.64(s,1h),8.34(s,1h),5.63(d,j=12.0hz,2h),5.08(dd,j=10.8,8.4hz,2h),3.69(s,2h),3.62(s,4h),2.65-2.53(m,2h),2.25(d,j=13.7hz,2h),2.05(d,j=14.1hz,2h),1.92-1.79(m,2h),1.67(d,j=11.6hz,6h),1.59-1.45(m,4h),1.42-1.30(m,8h),1.30-1.19(m,3h),1.00(d,j=10.3hz,2h),0.92(d,j=6.0hz,6h),0.60(dd,j=14.1,7.1hz,6h).
[0117]
13
c nmr(101mhz,dmso-d6)δ155.58,154.17,153.19,104.69,104.68,104.44,
103.82,99.49,91.57,91.49,81.68,81.58,71.17,71.00,66.51,53.86,52.74,51.56,51.54,45.67,36.80,36.71,36.01,34.23,31.67,31.30,26.21,24.94,20.81,20.74,20.54,20.49,13.67,13.46.
[0118]
ms(esi):[m+h]
+
=758.4
[0119]
实施例10化合物23的制备
[0120][0121]
氮气保护下,将化合物1(300mg,0.46mmol)溶解在50ml二氯甲烷中,室温下缓慢滴加四氢吡咯(4.0ml,47.8mmol),然后室温搅拌24小时。tlc监控反应完全后,反应液用饱和磷酸二氢钠水溶液、饱和食盐水洗涤,经无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1~40/1),得265mg白色固体产物23,收率78.5%。
[0122]1h nmr(400mhz,dmso-d6)δ8.58(s,1h),8.29(s,1h),5.63(d,j=15.0hz,2h),5.08(dd,j=11.2,4.3hz,2h),3.81(d,j=1.3hz,2h),2.59(s,5h),2.26(t,j=13.2hz,2h),2.05(d,j=14.3hz,2h),1.94-1.81(m,2h),1.78(s,4h),1.67(d,j=13.5hz,6h),1.55(s,4h),1.32(t,j=6.0hz,9h),1.28-1.21(m,3h),0.99(s,2h),0.92(d,j=6.3hz,6h),0.60(dd,j=10.9,7.2hz,6h).
[0123]
13
c nmr(101mhz,dmso-d6)δ156.17,153.50,153.03,104.68,104.67,104.06,103.76,100.94,91.58,91.49,81.69,81.59,71.19,71.03,53.27,51.58,51.55,51.19,45.69,36.80,36.71,36.02,34.24,31.69,31.28,26.23,26.21,24.94,23.67,20.82,20.69,20.54,20.49,13.68,13.51.
[0124]
ms(esi):[m+h]
+
=742.4
[0125]
实施例11化合物24的制备
[0126][0127]
氮气保护下,将化合物1(500mg,0.76mmol)溶解在100ml二氯甲烷中,缓慢滴加二甲胺的四氢呋喃溶液(2m,10.0ml,20.0mmol),然后室温搅拌24小时。tlc监控反应完全后,反应液用饱和磷酸二氢钠水溶液、饱和食盐水洗涤,经无水硫酸钠干燥,减压浓缩,残余物硅经胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1~50/1),得436mg淡黄色固体产物24,收率78.1%。
[0128]1h nmr(400mhz,dmso-d6)δ8.59(s,1h),8.30(s,1h),5.63(d,j=14.8hz,2h),5.07(dd,j=11.2,7.9hz,2h),3.63(s,4h),2.58(dd,j=11.4,4.5hz,2h),2.25(s,7h),2.05(d,
j=14.3hz,2h),1.92-1.78(m,2h),1.66(d,j=14.1hz,5h),1.60-1.42(m,4h),1.34(t,j=5.8hz,7h),1.27-1.18(m,2h),1.02(s,2h),0.92(d,j=6.3hz,5h),0.60(dd,j=9.2,7.4hz,5h).
[0129]1h nmr(400mhz,dmso-d6+d2o)δ5.61(d,j=14.7hz,2h),5.06(dd,j=11.1,8.7hz,2h),3.61(s,2h),2.56(dd,j=7.5,3.9hz,2h),2.24(s,7h),2.04(d,j=14.7hz,2h),1.85(s,2h),1.66(d,j=9.6hz,5h),1.51(d,j=22.1hz,4h),1.32(d,j=3.0hz,7h),1.23(d,j=5.2hz,2h),1.01(s,2h),0.91(d,j=6.3hz,5h),0.59(dd,j=9.4,7.3hz,5h).
[0130]
13
c nmr(101mhz,dmso-d6)δ156.13,153.92,153.21,104.68,104.18,103.86,100.40,91.59,91.49,81.69,81.59,78.91,71.17,71.03,66.80,51.57,51.54,45.68,44.01,36.79,36.70,36.01,34.23,33.38,32.02,31.71,31.56,31.27,30.30,29.90,26.20,24.94,20.92,20.81,20.64,20.54,20.49,13.66,13.52.
[0131]
ms(esi):[m+h]
+
=716.4
[0132]
实施例12化合物25的制备
[0133][0134]
氮气保护下,将化合物1(660mg,1.0mmol)溶解在100ml二氯甲烷中,在室温下缓慢滴加二乙胺(4.2ml,40.0mmol),然后加入多聚甲醛(30mg),室温继续搅拌2小时。tlc监控反应完全后,反应液用饱和磷酸二氢钠水溶液、饱和食盐水洗涤,有机相经无水硫酸钠干燥,减压浓缩,残余物硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=5/1~5/3),得333mg淡黄色固体产物25,收率44.8%。
[0135]1h nmr(400mhz,dmso-d6)δ8.58(s,1h),8.27(s,1h),5.62(d,j=16.3hz,2h),5.07(dd,j=11.1,8.1hz,2h),3.72(d,j=5.8hz,2h),2.66-2.53(m,5h),2.26(t,j=13.4hz,2h),2.05(d,j=14.5hz,2h),1.92-1.80(m,2h),1.67(d,j=13.1hz,6h),1.55(s,4h),1.33(d,j=4.6hz,8h),1.27(dd,j=14.2,10.5hz,4h),1.03(t,j=7.1hz,8h),0.92(d,j=6.3hz,6h),0.60(dd,j=9.7,7.3hz,6h).
[0136]
13
c nmr(101mhz,dmso-d6)δ156.18,153.96,153.00,105.21,104.74,104.67,104.13,103.91,101.15,91.59,91.49,81.70,81.59,71.18,71.02,58.67,57.25,56.17,51.57,50.74,46.31,45.69,36.80,36.72,36.02,34.25,31.68,31.24,26.22,26.21,24.95,20.82,20.54,20.50,13.68,13.60,13.47.
[0137]
ms(esi):[m+na]
+
=766.4
[0138]
实施例13化合物26的制备
[0139][0140]
氮气保护下,将化合物1(660mg,1.0mmol)溶解在100ml二氯甲烷中,室温下缓慢滴加二乙醇胺(1.0ml,10.4mmol),然后加入多聚甲醛(30mg),室温继续搅拌2小时。tlc监控反应完全后,反应液用饱和磷酸二氢钠水溶液、饱和食盐水洗涤,经无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1~20/1),得373mg淡黄色固体产物26,收率48.1%。
[0141]1h nmr(400mhz,cdcl3)δ8.60(s,1h),8.15(s,1h),5.43(d,j=14.0hz,2h),5.25(d,j=11.3hz,1h),5.14(d,j=11.2hz,1h),3.95(dd,j=35.6,14.1hz,2h),3.78(t,j=5.2hz,3h),2.96-2.67(m,5h),2.53-2.37(m,2h),2.25-2.03(m,11h),1.92(s,2h),1.78(d,j=12.5hz,3h),1.62(s,3h),1.54(d,j=14.4hz,2h),1.49-1.44(m,5h),1.39-1.28(m,3h),1.06(d,j=11.0hz,1h),1.00(d,j=6.1hz,5h),0.94-0.83(m,3h),0.72(dd,j=19.4,7.1hz,5h).
[0142]
13
c nmr(101mhz,dmso-d6)δ156.19,153.94,152.97,104.67,104.11,103.90,101.15,99.99,91.58,91.48,81.71,81.60,71.17,71.01,63.38,59.23,58.67,56.16,55.39,51.55,50.73,45.68,36.79,36.71,36.01,34.24,31.68,31.24,26.82,26.23,24.94,20.81,20.55,20.50,13.68,13.60.
[0143]
ms(esi):[m+h]
+
=776.4
[0144]
实施例14化合物27的制备
[0145][0146]
将化合物2(200mg,0.27mmol)溶解在四氢呋喃(10ml)和甲醇(10ml)的混合溶液中,再加入水合肼(80%,1.0ml,16.5mmol),室温搅拌过夜。tlc监控反应结束后,室温下减压浓缩,残余物用二氯甲烷溶解后,用饱和食盐水洗涤至有机相呈中性。有机相经无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=1/1~1/3),得166mg白色中间体27-1产物,收率84.1%。
[0147]
氮气保护下,将中间体27-1(100mg,0.14mmol)溶解在4ml干燥的dmf中,依次加入n,n-双芴甲氧羰基-l-赖氨酸(83mg,0.14mmol),hatu(2-(7-偶氮苯并三氮唑)-n,n,n',n'-四甲基脲六氟磷酸盐,107mg,0.28mmol),催化剂量的dmap(4-二甲氨基吡啶),室温下搅拌过夜。tlc监控反应结束后加入1m稀盐酸(10ml)淬灭反应,反应液用15ml乙酸乙酯萃取两次,合并有机相,依次用饱和碳酸氢钠水溶液、饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=2/1~1/1),得156mg白色固体产物27-2,收率85.5%。
[0148]
将中间体27-2(100mg,0.08mmol)溶解在dmf中(4ml),加入哌啶(1ml),室温搅拌1分钟,tlc监控至反应结束。反应液减压浓缩,残余物用ods柱纯化(80%~90%甲醇/水),得55.8mg白色固体产物27,收率84.4%。
[0149]1h nmr(400mhz,dmso-d6)δ8.57(s,2h),5.99(s,1h),5.62(d,j=32.5hz,2h),5.27(d,j=11.2hz,1h),5.09(d,j=11.2hz,1h),4.60(d,j=14.6hz,1h),4.41(d,j=14.7hz,1h),3.20(d,j=6.5hz,2h),2.64

2.54(m,3h),2.27(t,j=13.7hz,2h),2.05(d,j=13.7hz,2h),1.85(s,2h),1.66(s,6h),1.55(s,5h),1.34(d,j=6.7hz,13h),1.29

1.15(m,4h),1.02(s,2h),0.92(d,j=6.0hz,6h),0.60(dd,j=18.8,7.0hz,6h).
[0150]
13
c nmr(101mhz,dmso-d6)δ174.42,166.54,156.82,155.46,154.30,107.19,105.10,104.76,104.70,93.81,91.62,91.38,81.71,81.59,70.99,66.68,53.84,51.52,
45.65,41.78,40.62,37.01,36.75,35.99,35.86,34.22,33.17,31.61,31.15,26.23,26.09,24.95,22.97,20.80,20.66,20.52,13.62,13.56.
[0151]
ms(esi):[m+h]
+
=859.5
[0152]
实施例15化合物28的制备
[0153][0154]
氮气保护下,将中间体27-1(500mg,0.68mmol)溶解在20ml干燥的dmf中,加入fmoc-l-苯丙氨酸(271mg,0.68mmol),hatu(535mg,1.4mmol),催化剂量的dmap,在室温下搅拌过夜,tlc监控反应结束后,加入1m的稀盐酸(20ml)淬灭反应,用30ml乙酸乙酯萃取两次。合并有机相,依次用饱和碳酸氢钠水溶液、饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=2/1~1/1),得462mg白色固体产物28-1,收率61.7%。
[0155]
将中间体28-1(462mg,0.42mmol)溶解在dmf中(4ml),加入哌啶(1ml),室温下搅拌1分钟。tlc确认反应结束后,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=1/1~纯乙酸乙酯),得360mg白色固体产物28,收率97.6%。
[0156]1h nmr(400mhz,dmso-d6)δ8.57(d,j=4.8hz,2h),7.24(d,j=26.5hz,5h),5.99(s,1h),5.63(d,j=28.9hz,2h),5.27(d,j=11.1hz,1h),5.09(d,j=11.2hz,1h),4.62(d,j=14.6hz,1h),4.44(d,j=14.7hz,1h),3.56

3.47(m,1h),2.98(dd,j=13.3,4.8hz,1h),2.68-2.56(m,2h),2.27(t,j=12.9hz,2h),2.05(d,j=12.9hz,2h),1.85(s,2h),1.67(s,6h),1.56(d,j=6.4hz,2h),1.48(s,2h),1.41(s,2h),1.34(d,j=6.4hz,6h),1.30-1.20(m,3h),1.02(s,2h),0.92(d,j=5.8hz,6h),0.85(d,j=7.4hz,2h),0.61(dd,j=16.4,6.9hz,6h).
[0157]
13
c nmr(101mhz,dmso-d6)δ173.43,166.53,156.82,155.48,154.31,138.89,129.90,129.60,128.64,128.55,126.59,107.19,105.10,104.77,104.71,93.78,91.63,91.38,81.70,81.59,71.00,66.54,55.27,51.51,45.65,41.59,36.99,36.75,35.99,34.21,31.62,31.14,26.82,26.23,26.09,24.95,20.80,20.69,20.51,13.62,13.58.
[0158]
ms(esi):[m+h]
+
=878.4
[0159]
实施例16化合物29的制备
[0160][0161]
氮气保护下,将化合物1(527mg,0.80mmol)、特戊酸(98mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.40mmol)溶解在二氯甲烷(100ml)中,室温搅拌12小时。tlc检测反应完全后,将反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析进行纯化(洗脱剂:石油醚/乙酸乙酯=6/1~4/1),得300mg白色固体29,收率为50.5%。
[0162]1h nmr(400mhz,cdcl3)δ8.56(s,1h),8.27(s,1h),6.14(s,1h),5.42(s,1h),5.36(s,1h),5.26(d,j=11.4hz,1h),4.73(d,j=11.3hz,1h),2.85(dd,j=10.9,6.8hz,2h),2.43(ddd,j=14.2,7.4,3.6hz,2h),2.08

1.99(m,2h),1.96

1.85(m,2h),1.78(dd,j=21.8,8.4hz,4h),1.69

1.48(m,6h),1.42(t,j=8.8hz,9h),1.32(s,12h),0.97(d,j=6.0hz,6h),0.73(d,j=7.1hz,3h),0.62(d,j=7.2hz,3h).
[0163]
13
c nmr(101mhz,cdcl3)δ175.66,156.88,154.31,148.59,111.61,108.57,105.21,105.08,102.96,91.95,81.37,81.34,72.14,71.51,51.71,51.65,46.04,45.88,39.32,37.69,37.65,36.13,36.09,34.39,34.31,31.26,30.56,29.83,27.41,26.10,26.08,25.07,25.00,21.44,21.11,20.39,20.31,13.85,13.50.
[0164]
ms(esi):[m+h]
+
=743.4.
[0165]
实施例17化合物30的制备
[0166][0167]
氮气保护下,将化合物1(527mg,0.80mmol)、3,3-二甲基丁酸(111.5mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.40mmol)溶解在二氯甲烷(100ml)中,室温搅拌5小时。tlc检测反应完全后,将反应液用100ml饱和碳酸氢钠溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析进行纯化(洗脱剂:石油醚/乙酸乙酯=3/1),得330mg白色固体30,收率为54.5%。
[0168]1h nmr(400mhz,cdcl3)δ8.59(s,1h),8.26(s,1h),6.20(s,1h),5.42(s,1h),5.35(s,1h),5.26(d,j=11.4hz,1h),4.74(d,j=11.3hz,1h),2.91

2.75(m,2h),2.49

2.34(m,
4h),2.08

2.00(m,2h),1.90(dd,j=8.1,3.5hz,2h),1.81

1.70(m,4h),1.70

1.55(m,5h),1.43(d,j=7.4hz,8h),1.36

1.16(m,5h),1.11(s,9h),0.97(dd,j=6.0,2.4hz,6h),0.73(d,j=7.1hz,3h),0.61(d,j=7.2hz,3h).
[0169]
13
c nmr(101mhz,cdcl3)δ169.71,156.80,154.37,148.32,111.73,108.48,105.20,105.08,103.17,92.00,91.94,81.37,81.36,72.36,71.52,51.72,51.68,47.91,46.05,45.82,37.69,37.67,36.14,36.11,34.40,34.28,31.28,31.25,30.52,29.86,26.12,26.08,25.08,25.00,21.44,21.09,20.39,20.33,13.89,13.50.
[0170]
ms(esi):[m+h]
+
=757.4
[0171]
实施例18化合物31的制备
[0172][0173]
氮气保护下,将化合物1(790mg,1.2mmol)、苯甲酸(183.2mg,1.5mmol)、edci(690mg,3.6mmol)和dmap(73mg,0.60mmol)溶解在二氯甲烷(100ml)中,室温搅拌10小时。tlc检测反应结束后,将反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析进行纯化(洗脱剂:石油醚/乙酸乙酯=3/1),得510mg白色固体31,收率为55.7%。
[0174]1h nmr(400mhz,cdcl3)δ8.63(s,1h),8.35(s,1h),8.20

8.10(m,2h),7.63(dd,j=10.6,4.3hz,1h),7.51(t,j=7.7hz,2h),6.34(s,1h),5.44(s,1h),5.31(dd,j=12.9,6.0hz,3h),4.82(d,j=11.3hz,1h),2.86(ddd,j=11.0,7.2,3.9hz,2h),2.43(tdd,j=14.4,10.8,3.9hz,2h),2.12

1.98(m,2h),1.97

1.72(m,4h),1.70

1.51(m,7h),1.46(dd,j=12.0,3.8hz,7h),1.37

1.18(m,5h),0.98(d,j=6.1hz,3h),0.93(d,j=5.7hz,3h),0.77(d,j=7.2hz,3h),0.67(d,j=7.2hz,3h).
[0175]
13
c nmr(101mhz,cdcl3)δ164.12,156.99,154.41,148.38,133.64,130.17,129.79,128.80,111.95,108.64,105.20,105.12,103.40,91.99,81.39,81.35,72.42,71.56,51.74,51.64,46.07,45.78,37.71,37.53,36.16,36.10,34.41,34.20,31.29,30.65,26.12,25.09,24.99,21.46,20.82,20.41,20.28,14.00,13.55.
[0176]
ms(esi):[m+h]
+
=763.4
[0177]
实施例19化合物32的制备
[0178]
[0179]
氮气保护下,将化合物1(197.5mg,0.3mmol)、苯乙酰氯(69.6mg,0.45mmol)、edci(172mg,0.9mmol)和dmap(18mg,0.15mmol)溶解在二氯甲烷(100ml)中,室温搅拌24小时。tlc检测反应完全后,将反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析进行纯化(洗脱剂:石油醚/乙酸乙酯=3/1),得41mg白色固体32,收率为17.6%。
[0180]1h nmr(400mhz,cdcl3)δ8.60(s,1h),8.20(s,1h),7.47

7.27(m,5h),6.21(s,1h),5.41(s,1h),5.24(d,j=11.4hz,1h),5.10(s,1h),4.32(dd,j=8.9,4.8hz,1h),3.79(q,j=14.0hz,2h),2.81(ddd,j=10.6,7.1,3.1hz,1h),2.69(ddd,j=11.4,7.2,4.5hz,1h),2.41(dtd,j=18.0,14.1,3.8hz,2h),2.02(dd,j=10.3,4.2hz,2h),1.94

1.84(m,2h),1.79(d,j=9.3hz,2h),1.55(dd,j=54.8,10.4hz,7h),1.43(t,j=6.0hz,8h),1.27(dd,j=19.8,6.5hz,5h),0.97(d,j=5.9hz,6h),0.70(d,j=7.1hz,3h),0.38(d,j=7.1hz,3h).
[0181]
13
c nmr(101mhz,cdcl3)δ168.64,156.87,154.34,148.13,133.81,129.55,128.92,127.33,111.93,108.56,105.15,105.10,103.11,91.95,91.91,81.36,81.27,71.87,71.52,51.72,51.57,46.04,45.51,42.06,37.69,37.50,36.14,36.07,34.38,34.16,31.28,30.31,26.11,25.07,24.98,21.42,20.80,20.39,13.58,13.47.
[0182]
ms(esi):[m+h]
+
=777.4
[0183]
实施例20化合物33的制备
[0184][0185]
氮气保护下,将化合物1(460.8mg,0.7mmol)、胡椒酸(140mg,0.84)、edci(403mg,2.1mmol)和dmap(43mg,0.35mmol)溶解在二氯甲烷(100ml)中,室温搅拌24小时。tlc检测反应结束后,将反应液用100ml饱和碳酸氢钠溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析进行纯化(洗脱剂:石油醚/乙酸乙酯=3/1),得290mg白色固体33,收率为51.4%。
[0186]1h nmr(400mhz,cdcl3)δ8.61(s,1h),8.34(s,1h),7.76(dd,j=8.2,1.5hz,1h),7.55(d,j=1.3hz,1h),6.89(d,j=8.2hz,1h),6.31(s,1h),6.07(s,2h),5.43(s,1h),5.33(s,1h),5.31

5.27(m,1h),4.79(d,j=11.3hz,1h),2.90

2.81(m,2h),2.42(tdd,j=13.5,9.6,3.8hz,2h),2.04(d,j=14.4hz,2h),1.94

1.77(m,4h),1.70

1.54(m,7h),1.45(dd,j=15.6,4.5hz,8h),1.37

1.21(m,5h),0.96(dd,j=15.7,5.9hz,6h),0.75(d,j=7.1hz,3h),0.65(d,j=7.2hz,3h).
[0187]
13
c nmr(101mhz,cdcl3)δ163.47,156.95,154.37,152.31,148.41,148.16,126.11,123.64,111.86,109.95,108.66,108.34,105.18,105.10,103.39,102.12,91.98,81.38,81.34,72.40,71.54,51.73,51.63,46.06,45.77,37.69,37.53,36.14,36.09,34.40,34.21,31.28,30.64,26.11,25.08,24.98,21.44,20.84,20.39,20.28,14.00,
13.53.
[0188]
ms(esi):[m+h]
+
=807.4
[0189]
实施例21化合物34的制备
[0190][0191]
氮气保护下,将化合物1(790mg,1.2mmol)、2-吡啶甲酸(185mg,1.5mmol)、edci(690mg,3.6mmol)和dmap(73mg,0.60mmol)溶解在二氯甲烷(100ml)中,室温搅拌4小时。tlc检测反应完全后,将反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,将残余物经硅胶柱层析进行纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得700mg白色固体34,收率为76.4%。
[0192]1h nmr(400mhz,cdcl3)δ8.79(d,j=3.9hz,1h),8.63(d,j=2.6hz,1h),8.40(s,1h),8.20(d,j=7.8hz,1h),7.88(td,j=7.8,1.7hz,1h),7.52(ddd,j=7.6,4.7,1.0hz,1h),6.43(s,1h),5.43(s,1h),5.34(s,1h),5.29(t,j=5.7hz,1h),4.95(d,j=11.2hz,1h),2.83(ddd,j=11.0,9.0,5.6hz,2h),2.41(dt,j=25.4,7.4hz,2h),2.02(dd,j=8.9,4.8hz,2h),1.96

1.77(m,4h),1.67

1.48(m,7h),1.43(d,j=2.5hz,8h),1.38

1.23(m,5h),0.94(dd,j=17.3,5.8hz,6h),0.75(d,j=7.1hz,3h),0.66(d,j=7.1hz,3h).
[0193]
13
c nmr(101mhz,cdcl3)δ162.86,156.87,154.43,150.19,148.28,147.84,137.24,127.36,125.71,112.02,108.62,105.12,105.08,103.18,91.96,91.93,81.35,72.29,71.52,51.70,51.63,46.03,45.85,37.66,37.51,36.12,36.09,34.36,34.26,31.25,30.73,26.09,25.04,24.95,21.42,20.79,20.37,20.27,13.94,13.52.
[0194]
ms(esi):[m+h]
+
=764.4
[0195]
实施例22化合物35的制备
[0196][0197]
氮气保护下,将化合物1(527mg,0.8mmol)、2-吡啶乙酸盐酸盐(166.7mg,0.96mmol)、edci(460mg,2.4mmol)、dmap(49mg,0.4mmol)和三乙胺(0.5ml)溶于二氯甲烷(100ml),室温搅拌24小时。tlc检测反应完毕,反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓
缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得100mg白色固体35,收率为16.1%。
[0198]1h nmr(400mhz,cdcl3)δ8.61(s,1h),8.55(d,j=4.1hz,1h),8.27(s,1h),7.69(td,j=7.7,1.8hz,1h),7.40(d,j=7.8hz,1h),7.22(dd,j=7.0,5.4hz,1h),6.25(s,1h),5.41(s,1h),5.24(d,j=11.0hz,2h),4.60(d,j=11.3hz,1h),4.03(s,2h),2.88

2.67(m,2h),2.51

2.33(m,2h),2.11

1.98(m,2h),1.90(d,j=10.4hz,2h),1.76(dd,j=30.4,11.5hz,3h),1.60(s,10h),1.55

1.48(m,3h),1.43(s,6h),0.98(t,j=6.5hz,6h),0.71(d,j=7.1hz,3h),0.46(d,j=7.2hz,3h).
[0199]
13
c nmr(101mhz,cdcl3)δ167.87,156.76,154.30,154.19,149.61,148.09,136.95,124.15,122.39,111.91,108.46,105.16,105.09,102.99,91.90,81.35,81.30,72.08,71.47,51.64,51.55,45.96,45.61,44.41,37.64,37.59,36.08,36.03,34.32,34.20,31.22,30.44,26.09,25.02,24.95,21.39,21.09,20.39,13.65,13.48.
[0200]
ms(esi):[m+h]
+
=778.4
[0201]
实施例23化合物36的制备
[0202][0203]
氮气保护下,将化合物1(527mg,0.8mmol)、2-萘甲酸(165.3mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.4mmol)溶于二氯甲烷(100ml),室温搅拌15小时。tlc检测反应结束后,将反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤、无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=3/1),得400mg白色固体36,收率为61.5%。
[0204]1h nmr(400mhz,cdcl3)δ8.74(s,1h),8.65(s,1h),8.37(s,1h),8.15(dd,j=8.6,1.4hz,1h),8.05

7.88(m,3h),7.61(dt,j=14.9,6.9hz,2h),6.41(s,1h),5.45(s,1h),5.32(d,j=15.6hz,2h),4.89(d,j=11.2hz,1h),2.88(dd,j=8.7,6.1hz,2h),2.44(qd,j=13.7,3.7hz,2h),2.10

1.99(m,2h),1.87(ddd,j=34.7,20.1,8.2hz,4h),1.67

1.55(m,7h),1.48(dd,j=22.3,5.7hz,8h),1.39

1.14(m,5h),0.99(d,j=6.0hz,3h),0.91(d,j=5.1hz,3h),0.78(d,j=7.1hz,3h),0.72(d,j=7.1hz,3h).
[0205]
13
c nmr(101mhz,cdcl3)δ164.29,157.05,154.45,148.48,135.96,132.74,131.96,129.60,128.78,128.64,128.01,127.04,127.01,125.53,111.99,108.69,105.22,105.15,103.47,92.02,81.41,81.36,72.47,71.59,51.76,51.63,46.09,45.79,37.73,37.52,36.17,36.11,34.43,34.15,31.32,30.68,26.14,25.11,24.99,21.48,20.89,20.42,20.26,14.09,13.58.
[0206]
ms(esi):[m+h]
+
=813.4
[0207]
实施例24化合物37的制备
[0208][0209]
氮气保护下,将化合物1(527mg,0.8mmol)、1-萘甲酸(165.3mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.4mmol)溶解在二氯甲烷(100ml)中,室温搅拌15小时。tlc检测反应完全后,反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=3/1),得469mg白色固体37,收率为72.2%。
[0210]1h nmr(400mhz,cdcl3)δ9.04(d,j=8.6hz,1h),8.67(s,1h),8.42

8.32(m,2h),8.10(d,j=8.2hz,1h),7.92(d,j=8.0hz,1h),7.65(ddd,j=8.5,6.9,1.3hz,1h),7.61

7.52(m,2h),6.42(s,1h),5.45(s,1h),5.32(t,j=5.7hz,2h),4.87(d,j=11.3hz,1h),2.90(ddd,j=10.9,7.2,3.9hz,2h),2.43(qd,j=14.5,3.8hz,2h),2.11

1.99(m,2h),1.95

1.74(m,4h),1.67

1.51(m,7h),1.45(d,j=1.5hz,8h),1.31(ddd,j=21.3,13.7,8.6hz,5h),0.99(d,j=6.1hz,3h),0.91(d,j=5.3hz,3h),0.79(d,j=7.1hz,3h),0.71(d,j=7.2hz,3h).
[0211]
13
c nmr(101mhz,cdcl3)δ164.78,157.08,154.51,148.55,134.36,134.15,131.96,130.77,128.75,128.40,126.64,126.17,126.04,124.63,112.02,108.86,105.20,105.13,103.48,92.00,81.40,81.36,72.51,71.60,51.75,51.63,46.09,45.78,37.72,37.50,36.17,36.10,34.42,34.16,31.33,30.64,26.13,25.10,24.98,21.47,20.84,20.41,20.26,14.07,13.56.
[0212]
ms(esi):[m+h]
+
=813.4
[0213]
实施例25化合物38的制备
[0214][0215]
氮气保护下,将化合物1(527mg,0.8mmol)、2-萘乙酸(178mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.4mmol)溶于二氯甲烷(100ml)中,室温搅拌16小时。tlc检测反应完全后,将反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化
(洗脱剂:石油醚/乙酸乙酯=5/1),得346mg白色固体38,收率为52.3%。
[0216]1h nmr(400mhz,cdcl3)δ8.63(s,1h),8.14(s,1h),7.88

7.81(m,4h),7.56

7.45(m,3h),6.26(s,1h),5.40(s,1h),5.23(d,j=11.3hz,1h),4.77(s,1h),4.14(d,j=11.0hz,1h),3.96(dd,j=37.7,13.7hz,2h),2.80(ddd,j=10.7,7.1,3.3hz,1h),2.57(ddd,j=11.3,7.1,4.5hz,1h),2.43(td,j=14.1,3.8hz,1h),2.31(td,j=14.1,3.8hz,1h),2.01(dt,j=18.8,3.5hz,2h),1.97

1.83(m,2h),1.82

1.51(m,7h),1.45(dd,j=33.3,12.1hz,8h),1.35

1.11(m,5h),1.04(d,j=3.7hz,2h),0.96(d,j=6.0hz,3h),0.77(d,j=5.7hz,3h),0.68(d,j=7.1hz,3h),0.27(d,j=7.1hz,3h).
[0217]
13
c nmr(101mhz,cdcl3)δ168.52,156.88,154.32,148.02,133.78,132.77,131.38,128.58,128.06,127.81,127.56,126.62,126.15,111.96,108.66,105.08,105.01,103.17,91.93,91.79,81.33,81.15,71.68,71.50,51.70,51.34,46.01,45.33,42.26,37.65,37.12,36.13,36.00,34.35,33.80,31.28,30.14,26.10,26.08,25.05,24.83,21.38,20.36,20.22,20.04,13.48,13.44.
[0218]
ms(esi):[m+h]
+
=827.4
[0219]
实施例26化合物39的制备
[0220][0221]
氮气保护下,将化合物1(527mg,0.8mmol)、1-萘乙酸(178mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.4mmol)溶于二氯甲烷(100ml)中,室温搅拌16小时。tlc检测反应完全后,将反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸钠,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=5/1),得160mg白色固体39,收率为24.2%。
[0222]1h nmr(400mhz,cdcl3)δ8.60(s,1h),8.20(d,j=8.4hz,1h),8.13(s,1h),7.83(dd,j=22.6,8.1hz,2h),7.52(ddd,j=22.0,15.1,8.2hz,4h),6.24(s,1h),5.39(s,1h),5.21(d,j=11.3hz,1h),4.68(s,1h),4.23(q,j=14.4hz,2h),3.91(d,j=11.3hz,1h),2.87

2.71(m,1h),2.52

2.29(m,3h),1.94

1.63(m,5h),1.59(s,4h),1.46

1.28(m,11h),1.24

0.99(m,5h),0.95(t,j=5.6hz,7h),0.66(d,j=7.1hz,3h),0.05(d,j=7.1hz,3h).
[0223]
13
c nmr(101mhz,cdcl3)δ168.79,156.83,154.27,148.12,134.18,132.34,130.28,128.78,128.71,128.21,126.91,126.08,125.80,124.27,111.91,108.63,105.09,105.04,103.01,91.93,91.69,81.34,81.16,71.73,71.51,51.71,51.47,46.02,45.15,40.19,37.67,37.30,36.14,36.02,34.36,34.22,31.28,29.97,26.11,26.09,25.06,24.91,21.40,20.38,20.36,19.86,13.43,13.20.
[0224]
ms(esi):[m+h]
+
=827.4
[0225]
实施例27化合物40的制备
[0226][0227]
氮气保护下,将化合物1(527mg,0.8mmol)、7-喹啉甲酸(166mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.4mmol)溶于二氯甲烷(100ml)中,室温搅拌16小时。tlc检测反应完全后,反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸钠,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得300mg白色固体40,收率为46.1%。
[0228]1h nmr(400mhz,cdcl3)δ9.03(dd,j=4.2,1.6hz,1h),8.94(s,1h),8.66(s,1h),8.40(s,1h),8.24(d,j=8.5hz,2h),7.95(d,j=8.6hz,1h),7.54(dd,j=8.3,4.2hz,1h),6.40(s,1h),5.45(s,1h),5.34(d,j=9.7hz,2h),4.90(d,j=11.2hz,1h),2.88(ddd,j=11.1,7.2,3.8hz,2h),2.42(tt,j=13.5,6.9hz,2h),2.07

2.01(m,2h),1.91

1.75(m,4h),1.61(dd,j=31.2,11.6hz,7h),1.45(d,j=4.1hz,8h),1.34

1.27(m,5h),0.98(d,j=6.1hz,3h),0.90(d,j=5.5hz,3h),0.78(d,j=7.1hz,3h),0.72(d,j=7.2hz,3h).
[0229]
13
c nmr(101mhz,cdcl3)δ163.91,157.04,154.48,151.74,148.36,147.85,136.00,132.76,131.23,130.51,128.64,126.49,123.27,112.12,108.68,105.19,105.12,103.34,92.00,81.39,81.37,72.48,71.57,51.75,51.63,46.08,45.78,37.71,37.48,36.16,36.10,34.41,34.16,31.30,30.73,29.83,26.13,25.09,24.98,21.46,20.85,20.40,20.25,14.11,13.56.
[0230]
ms(esi):[m+h]
+
=814.4
[0231]
实施例28化合物41的制备
[0232][0233]
氮气保护下,将化合物1(527mg,0.8mmol)、8-喹啉甲酸(166mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.4mmol)溶于二氯甲烷(100ml)中,室温搅拌16小时。tlc检测反应完全后,反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得200mg白色固体41,收率为30.7%。
[0234]1h nmr(400mhz,cdcl3)δ8.98(d,j=3.2hz,1h),8.64(s,1h),8.39(s,1h),8.21(d,
j=8.3hz,1h),8.12(d,j=7.1hz,1h),7.99(d,j=8.2hz,1h),7.62(t,j=7.6hz,1h),7.48(dd,j=8.1,4.1hz,1h),6.48(s,1h),5.42(d,j=14.2hz,2h),5.29(s,1h),5.19(d,j=11.2hz,1h),2.96

2.83(m,2h),2.50

2.37(m,2h),2.04(d,j=14.2hz,2h),1.96

1.74(m,5h),1.64(dd,j=16.7,9.0hz,7h),1.45(d,j=2.7hz,8h),1.38

1.20(m,4h),0.96(dd,j=10.0,6.0hz,6h),0.75(dd,j=18.6,7.0hz,6h).
[0235]
13
c nmr(101mhz,cdcl3)δ166.07,156.97,154.53,151.26,148.87,145.80,136.30,131.89,131.68,130.67,128.61,125.86,121.84,111.98,109.20,105.14,105.10,103.32,92.03,91.99,81.46,81.40,72.56,71.60,51.74,51.69,46.08,45.95,37.70,37.65,36.15,34.41,34.30,31.33,30.69,26.15,26.12,25.08,25.03,21.46,21.10,20.40,20.37,13.95,13.54.
[0236]
ms(esi):[m+h]
+
:814.4
[0237]
实施例29化合物42的制备:
[0238][0239]
在氩气保护下,化合物1(0.20g,0.30mmol)溶解于无水dcm(5ml)中,滴加tea(0.20ml,1.4mmol),反应体系降温至0℃,缓慢滴加癸酰氯(189μl,0.90mmol,用5ml dcm稀释),滴加结束后,在冰水浴冷却下持续搅拌1小时。tlc监控反应结束后,加入甲醇(1ml)淬灭反应,减压蒸干溶剂,残余物经硅胶柱纯化(洗脱剂:石油醚/乙酸乙酯=5/1),得122mg白色固体产物42,收率50.0%。
[0240]1h nmr(400mhz,cdcl3)δ8.62(s,1h),8.30(s,1h),6.23(s,1h),5.45(s,1h),5.39(s,1h),5.28(d,j=11.3hz,1h),4.75(d,j=11.3hz,1h),2.93-2.79(m,2h),2.49(dd,j=17.8,10.4hz,4h),2.13-2.01(m,2h),2.00-1.88(m,2h),1.84-1.71(m,6h),1.68-1.61(m,4h),1.58-1.51(m,2h),1.46(d,j=6.2hz,8h),1.40(dd,j=9.7,4.8hz,4h),1.30(d,j=4.2hz,10h),1.09(dd,j=22.1,9.7hz,2h),1.00(dd,j=6.0,2.6hz,6h),0.91(t,j=6.8hz,3h),0.76(d,j=7.1hz,3h),0.63(d,j=7.2hz,3h).
[0241]
13
c nmr(101mhz,cdcl3)δ170.99,156.68,154.18,148.11,111.59,108.26,105.08,104.97,103.05,91.84,91.80,81.23,81.19,72.13,71.37,51.54,51.48,45.87,45.62,37.54,37.52,35.98,35.94,34.58,34.23,34.10,31.90,31.11,30.40,29.48,29.39,29.34,29.29,25.98,25.22,24.93,24.86,22.69,21.29,20.95,20.28,20.21,14.13,13.76,13.40.
[0242]
ms(esi):[m+h]
+
=813.5
[0243]
实施例30化合物43的制备
[0244][0245]
氮气保护下,将化合物1(527mg,0.8mmol)、硬脂酸(246mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.4mmol)溶于二氯甲烷(100ml),室温搅拌4小时。tlc检测反应完全后,反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:纯二氯甲烷),得360mg白色固体43,收率为50.2%。
[0246]1h nmr(400mhz,cdcl3)δ8.60(s,1h),8.27(s,1h),6.20(s,1h),5.42(s,1h),5.36(s,1h),5.26(d,j=11.4hz,1h),4.72(d,j=11.3hz,1h),2.83(ddd,j=11.1,7.2,4.2hz,2h),2.51

2.38(m,4h),2.09

2.01(m,2h),1.95

1.87(m,2h),1.75(t,j=10.1hz,6h),1.60(d,j=13.0hz,9h),1.44(d,j=6.2hz,8h),1.28(d,j=22.6hz,22h),1.15

0.92(m,9h),0.88(t,j=6.8hz,3h),0.73(d,j=7.1hz,3h),0.60(d,j=7.2hz,3h).
[0247]
13
c nmr(101mhz,cdcl3)δ171.14,156.81,154.31,148.24,111.73,108.40,105.22,105.11,103.19,91.97,91.93,81.37,81.33,72.26,71.51,51.67,51.60,46.00,45.74,37.68,37.65,36.11,36.07,34.72,34.36,34.24,32.07,31.24,30.53,29.85,29.81,29.68,29.53,29.51,29.44,26.12,25.37,25.06,24.99,22.84,21.43,21.08,20.42,20.35,14.28,13.89,13.53.
[0248]
ms(esi):[m+h]
+
=897.6
[0249]
实施例31化合物44的制备
[0250][0251]
氮气保护下,将化合物1(527mg,0.8mmol)、软脂酸(273mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.4mmol)溶于二氯甲烷(100ml),室温搅拌5小时。tlc检测反应完全后,反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷),得380mg白色固体44,收率为51.4%。
[0252]1h nmr(400mhz,cdcl3)δ8.59(s,1h),8.27(s,1h),6.20(s,1h),5.42(s,1h),5.36(s,1h),5.26(d,j=11.3hz,1h),4.72(d,j=11.3hz,1h),2.82(dd,j=9.2,5.9hz,2h),2.51

2.38(m,4h),2.04(dd,j=14.4,2.7hz,2h),1.90(s,2h),1.83

1.69(m,6h),1.60(d,j=14.4hz,8h),1.43(d,j=6.2hz,8h),1.28(d,j=23.0hz,27h),1.15

0.93(m,9h),0.87(t,j=6.8hz,3h),0.73(d,j=7.1hz,3h),0.60(d,j=7.1hz,3h).
[0253]
13
c nmr(101mhz,cdcl3)δ171.14,156.81,154.31,148.24,111.73,108.40,105.22,105.11,103.19,91.97,91.93,81.37,81.33,72.26,71.51,51.67,51.60,45.99,45.74,37.68,37.65,36.10,36.07,34.72,34.36,34.23,32.07,31.24,30.52,29.85,29.81,29.68,29.53,29.51,29.44,26.11,25.36,25.06,24.99,22.84,21.42,21.08,20.42,20.35,14.27,13.89,13.53.
[0254]
ms(esi):[m+h]
+
=925.6
[0255]
实施例32化合物45的制备
[0256][0257]
氮气保护下,将化合物1(527mg,0.8mmol)、亚油酸(269mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.4mmol)溶于二氯甲烷(100ml),室温搅拌8小时。tlc检测反应完全后,反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1),得330mg白色固体45,收率为44.8%。
[0258]1h nmr(400mhz,cdcl3)δ8.60(s,1h),8.28(s,1h),6.20(s,1h),5.55

5.20(m,7h),4.72(d,j=11.2hz,1h),2.79(dd,j=18.2,11.9hz,4h),2.57

2.34(m,4h),2.05(d,j=6.8hz,6h),1.90(s,2h),1.84

1.68(m,6h),1.60(d,j=14.9hz,8h),1.48

1.29(m,22h),1.02(dd,j=33.9,7.9hz,8h),0.89(t,j=6.6hz,3h),0.73(d,j=7.0hz,3h),0.60(d,j=7.1hz,3h).
[0259]
13
c nmr(101mhz,cdcl3)δ171.10,156.81,154.31,148.23,130.41,130.10,128.27,128.01,111.74,108.39,105.22,105.11,103.19,91.97,91.93,81.37,81.32,72.26,71.51,51.67,51.60,45.99,45.74,37.68,37.65,36.10,36.06,34.68,34.36,34.23,31.67,31.23,30.52,29.80,29.49,29.42,29.38,29.30,27.35,27.34,26.11,25.78,25.32,25.06,24.98,22.72,21.42,21.07,20.41,20.35,14.23,13.89,13.53.
[0260]
ms(esi):[m+h]
+
=921.6
[0261]
实施例33化合物46的制备
[0262][0263]
氮气保护下,将化合物1(527mg,0.8mmol)、油酸(271mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.4mmol)溶于二氯甲烷(100ml),室温搅拌6小时。tlc检测反应完全后,反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2)。合并有机
相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1),得390mg白色固体46,收率为52.8%。
[0264]1h nmr(400mhz,cdcl3)δ8.60(s,1h),8.27(s,1h),6.20(s,1h),5.42(s,1h),5.39

5.31(m,3h),5.26(d,j=11.4hz,1h),4.72(d,j=11.3hz,1h),2.86

2.77(m,2h),2.51

2.38(m,4h),2.03(t,j=9.5hz,6h),1.95

1.88(m,2h),1.83

1.70(m,6h),1.60(d,j=14.3hz,8h),1.43(d,j=6.1hz,8h),1.30(dd,j=16.0,9.0hz,19h),1.17

0.92(m,9h),0.88(dd,j=8.8,4.7hz,3h),0.73(d,j=7.1hz,3h),0.60(d,j=7.2hz,3h).
[0265]
13
c nmr(101mhz,cdcl3)δ171.11,156.81,154.31,148.23,130.23,129.80,111.74,108.39,105.22,105.11,103.19,91.97,91.93,81.37,81.32,72.26,71.51,51.67,51.60,46.00,45.74,37.68,37.65,36.11,36.07,34.69,34.36,34.24,32.05,31.24,30.52,29.91,29.90,29.68,29.48,29.47,29.42,29.40,29.30,27.38,27.32,26.11,25.33,25.06,24.99,22.83,21.42,21.08,20.42,20.35,14.27,13.89,13.53.
[0266]
ms(esi):[m+h]
+
=923.6
[0267]
实施例34化合物47的制备
[0268][0269]
氮气保护下,将化合物1(527mg,0.8mmol)、亚麻酸(267mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.4mmol)溶于二氯甲烷(100ml),室温搅拌8小时。tlc检测反应完全后,反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1),得220mg白色固体47,收率为29.9%。
[0270]1h nmr(400mhz,cdcl3)δ8.60(s,1h),8.27(s,1h),6.20(s,1h),5.47

5.28(m,8h),5.26(d,j=11.4hz,1h),4.72(d,j=11.3hz,1h),2.91

2.71(m,6h),2.51

2.38(m,4h),2.06(dt,j=14.4,7.6hz,6h),1.90(s,2h),1.83

1.70(m,6h),1.60(d,j=11.4hz,8h),1.43(d,j=6.1hz,8h),1.38

1.29(m,10h),1.02

0.93(m,9h),0.73(d,j=7.1hz,3h),0.60(d,j=7.1hz,3h).
[0271]
13
c nmr(101mhz,cdcl3)δ171.10,156.81,154.31,148.23,132.12,130.32,128.46,128.36,127.95,127.24,111.74,108.39,105.22,105.11,103.18,91.96,91.93,81.37,81.32,72.26,71.51,51.67,51.60,45.99,45.74,37.68,37.65,36.10,36.06,34.67,34.36,34.24,31.23,30.52,29.77,29.42,29.38,29.29,27.35,26.11,25.76,25.67,25.32,25.06,24.99,21.42,21.07,20.70,20.41,20.35,14.43,13.89,13.53.
[0272]
ms(esi):[m+h]
+
=919.6
[0273]
实施例35化合物48的制备
[0274][0275]
氮气保护下,将化合物1(527mg,0.8mmol)、4-(2-萘基)-4-氧代丁酸(219mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.4mmol)溶于二氯甲烷(100ml),室温搅拌22小时。tlc检测反应完全后,将反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2)。合并有机相,用饱和食盐水洗涤,经无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得250mg白色固体48,收率为36.0%。
[0276]1h nmr(400mhz,cdcl3)δ8.62(s,1h),8.49(s,1h),8.36(s,1h),8.02

7.94(m,2h),7.92

7.84(m,2h),7.63

7.53(m,2h),6.24(s,1h),5.44(s,1h),5.39(s,1h),5.26(d,j=11.3hz,1h),4.93(d,j=11.2hz,1h),3.65

3.43(m,2h),3.04

2.91(m,2h),2.84(ddd,j=13.8,9.0,5.7hz,2h),2.42(td,j=14.0,3.6hz,2h),2.07

1.98(m,2h),1.94

1.85(m,2h),1.84

1.66(m,6h),1.60(dd,j=11.0,4.2hz,3h),1.56

1.48(m,2h),1.42(d,j=4.3hz,8h),1.36

1.28(m,3h),0.96(t,j=6.1hz,6h),0.73(d,j=7.1hz,3h),0.64(d,j=7.1hz,3h).
[0277]
13
c nmr(101mhz,cdcl3)δ197.89,170.62,156.85,154.42,148.37,135.80,133.92,132.62,129.87,129.69,128.67,128.60,127.89,126.98,123.73,111.85,108.91,105.05,105.02,103.04,92.04,91.89,81.50,81.32,72.26,71.50,51.70,51.65,45.99,45.91,37.63,37.60,36.09,34.38,34.31,33.61,31.26,30.65,28.57,26.10,26.06,25.00,21.35,20.98,20.33,13.75,13.46.
[0278]
ms(esi):[m+h]
+
=869.4
[0279]
实施例36化合物49的制备
[0280][0281]
氮气保护下,将化合物1(527mg,0.8mmol)、4-(1-萘基)-4-氧代丁酸(219mg,0.96mmol)、edci(460mg,2.4mmol)和dmap(49mg,0.4mmol)溶于二氯甲烷(100ml),室温搅拌6小时。tlc检测反应完全后,将反应液用100ml饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得512mg白色固体49,收率为
73.7%。
[0282]1h nmr(400mhz,cdcl3)δ8.60(d,j=9.2hz,2h),8.35(s,1h),8.09

7.93(m,2h),7.88(d,j=7.6hz,1h),7.63

7.45(m,3h),6.21(s,1h),5.41(s,1h),5.33(s,1h),5.27(d,j=11.3hz,1h),4.93(d,j=11.2hz,1h),3.59(ddd,j=18.0,8.5,4.4hz,1h),3.36(dd,j=13.2,5.2hz,1h),3.13

3.00(m,1h),3.00

2.89(m,1h),2.83(s,2h),2.41(dd,j=27.8,13.6hz,2h),2.02(t,j=12.6hz,2h),1.96

1.70(m,6h),1.57(dd,j=23.1,11.5hz,6h),1.42(s,8h),1.25(dd,j=19.3,9.1hz,3h),0.97(d,j=5.7hz,4h),0.88(d,j=5.6hz,3h),0.74(d,j=7.0hz,3h),0.65(d,j=7.0hz,3h).
[0283]
13
c nmr(100mhz,cdcl3)δ202.03,170.72,156.92,154.52,148.44,135.39,134.13,133.09,130.31,128.54,128.07,126.62,125.97,124.61,111.97,109.08,105.08,105.03,103.03,92.01,91.95,81.55,81.38,72.34,71.56,51.72,51.69,46.05,45.96,37.68,37.41,36.55,36.14,34.38,34.33,31.31,30.67,28.87,26.15,26.12,25.07,24.98,21.43,20.95,20.39,20.32,13.79,13.51.
[0284]
ms(esi):[m+h]
+
=869.4
[0285]
实施例37化合物50的制备
[0286][0287]
氮气保护下,在反应瓶中依次加入化合物1(1g,1.52mmol)、5-氧代-5-(2-(三甲基甲硅烷基)乙氧基)丁酸(436mg,2mmol)、edci(573mg,3mmol)、hobt(405mg,3mmol)、dmap(488mg,4mmol)、dmf(10ml)和dcm(40ml),搅拌至完全溶解,慢慢升温至35℃并继续搅拌5小时,tlc监控至反应完全。将反应液倒入100ml纯水中,加入二氯甲烷100ml,进行分液萃取,水相用二氯甲烷萃取(50ml
×
2)。合并有机相,依次用10%柠檬酸水溶液、饱和碳酸氢钠水溶液、饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=20/1),得890mg白色泡沫状固体50,收率约68.4%。
[0288]1h nmr(400mhz,cdcl3)δ8.60(s,1h),8.31(s,1h),6.19(s,1h),5.42(s,2h),5.26(d,j=11.4hz,1h),4.81(d,j=11.3hz,1h),4.17(t,j=8.5hz,2h),2.88

2.59(m,6h),2.50

2.36(m,2h),2.04(d,j=14.4hz,2h),1.96

1.85(m,2h),1.84

1.69(m,4h),1.58(s,10h),1.43(d,j=3.3hz,6h),1.36

1.29(m,2h),1.11

1.02(m,2h),0.97(d,j=6.1hz,6h),0.73(d,j=7.1hz,3h),0.61(d,j=7.1hz,3h),0.04(s,9h).
[0289]
ms(esi):[m+h]
+
=859.4
[0290]
实施例38化合物51的制备
[0291][0292]
氮气保护下,将化合物1(1g,1.52mmol,1.0eq)、5-氧代-5-(2-(三甲基甲硅烷基)乙氧基)戊酸(465mg,2mmol)、edci(573mg,3mmol)、hobt(405mg,3mmol)、dmap(488mg,4mmol)溶于dmf(10ml)和dcm(40ml)中,在35℃搅拌5小时,tlc检测反应完全后,反应液中加入100ml纯水和100ml二氯甲烷进行搅拌。分离有机相后,水相继续用二氯甲烷萃取(50ml
×
2)。合并有机相,依次用10%柠檬酸水溶液、饱和碳酸氢钠水溶液、饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=15/1),得988mg白色固体51-1,收率74.5%。
[0293]1h nmr(400mhz,cdcl3)δ8.60(s,1h),8.29(s,1h),6.21(s,1h),5.40(d,j=14.4hz,2h),5.25(d,j=11.4hz,1h),4.72(d,j=11.3hz,1h),4.26

4.07(m,2h),2.82(dd,j=10.9,6.9hz,2h),2.57(t,j=7.1hz,2h),2.50

2.35(m,4h),2.04(dd,j=14.4,7.1hz,4h),1.89(s,2h),1.78(dd,j=19.7,8.9hz,4h),1.67

1.24(m,16h),1.13

0.91(m,10h),0.73(d,j=7.1hz,3h),0.59(d,j=7.1hz,3h),0.04(s,9h).
[0294]
ms(esi):[m+h]
+
=873.4
[0295]
氮气保护下,将化合物51-1(600mg,0.69mmol)溶于四氢呋喃(15ml),降温至0℃后加入tbaf(1.8g,6.9mmol),加完后慢慢升温至20℃并在该温度下下搅拌2小时。tlc检测反应完全后,将反应液减压浓缩,向残余物中加入100ml纯水和100ml二氯甲烷进行搅拌。分离有机相,水相用二氯甲烷萃取(50ml
×
2)。合并有机相,依次用10%柠檬酸水溶液、饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=40/1),得293mg白色固体51,收率54.9%。
[0296]1h nmr(400mhz,cdcl3)δ:8.61(s,1h),8.29(s,1h),6.21(s,1h),5.41(d,j=12.0hz,2h),5.26(d,j=11.2hz,1h),4.73(d,j=11.2hz,1h),2.82(s,2h),2.60(t,j=7.0hz,2h),2.46(dt,j=28.2,10.4hz,4h),2.06(dt,j=14.1,7.1hz,4h),1.89(s,2h),1.84

1.72(m,4h),1.67

1.22(m,17h),1.13

0.87(m,8h),0.73(d,j=7.0hz,3h),0.60(d,j=7.0hz,3h).
[0297]
13
c nmr(101mhz,cdcl3)δ178.19,170.26,156.79,154.34,147.99,111.87,108.35,105.22,105.11,103.10,91.97,91.92,81.36,72.22,71.49,51.64,51.59,45.97,45.70,37.65,37.57,36.08,36.05,34.33,34.19,33.39,32.96,31.23,30.52,26.10,25.03,24.96,21.40,20.97,20.39,20.34,20.03,13.89,13.51.
[0298]
ms(esi):[m+h]
+
=773.4
[0299]
实施例39化合物52的制备
[0300][0301]
氮气保护下,将化合物1(1g,1.52mmol,1.0eq)、6-氧代-6-(2-(三甲基甲硅烷基)乙氧基)己酸(500mg,2.03mmol)、edci(573mg,3mmol)、hobt(405mg,3mmol)、dmap(488mg,4mmol)溶于dmf(10ml)和dcm(40ml)中,在35℃搅拌5小时。tlc检测反应完全后,反应液中加入100ml纯水和100ml二氯甲烷进行搅拌。分离有机相,水相用二氯甲烷萃取两次(50ml
×
2)。合并有机相,依次用10%柠檬酸水溶液、饱和碳酸氢钠水溶液、饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=15/1),得1.07g白色固体52-1,收率79.6%。
[0302]1h nmr(400mhz,cdcl3)δ8.59(s,1h),8.29(s,1h),6.20(s,1h),5.40(d,j=14.7hz,2h),5.25(d,j=11.4hz,1h),4.71(d,j=11.3hz,1h),4.25

4.08(m,2h),2.83(dd,j=13.4,9.0hz,2h),2.58

2.27(m,6h),2.09

1.98(m,2h),1.96

1.85(m,2h),1.85

1.67(m,8h),1.66

1.22(m,16h),1.16

0.90(m,10h),0.73(d,j=7.1hz,3h),0.59(d,j=7.2hz,3h),0.04(s,9h).
[0303]
ms(esi):[m+h]
+
=887.5
[0304]
氮气保护下,将化合物52-1(600mg,0.67mmol)溶于四氢呋喃(15ml),降温至0℃后加入tbaf(1.8g,6.9mmol),加完后慢慢升温至20℃并在该温度下下搅拌2小时。tlc检测反应完全后,反应液减压浓缩,残余物中加入100ml纯水和100ml二氯甲烷进行搅拌。分离有机相,水相用二氯甲烷萃取两次(50ml
×
2)。合并有机相,依次用10%柠檬酸水溶液、饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=40/1),得260mg白色固体52,收率49.2%。
[0305]1h nmr(400mhz,cdcl3)δ8.61(s,1h),8.29(s,1h),6.20(s,1h),5.40(d,j=13.6hz,2h),5.26(d,j=11.4hz,1h),4.72(d,j=11.3hz,1h),2.87

2.77(m,2h),2.53(t,j=6.8hz,2h),2.48

2.37(m,4h),2.04(dd,j=14.3,2.7hz,2h),1.89(s,2h),1.77(dd,j=17.4,11.3hz,8h),1.68

1.18(m,16h),1.16

0.84(m,9h),0.73(d,j=7.1hz,3h),0.59(d,j=7.1hz,3h).
[0306]
13
c nmr(101mhz,cdcl3)δ178.64,170.64,156.78,154.32,148.11,111.80,108.35,105.22,105.10,103.11,91.94,91.91,81.36,81.34,72.24,71.48,51.64,51.58,45.97,45.70,37.65,37.55,36.08,36.04,34.33,34.18,34.14,33.61,31.22,30.50,26.09,25.03,24.95,24.46,24.22,21.40,20.99,20.39,20.33,13.89,13.51.
[0307]
ms(esi):[m+h]
+
=787.4
[0308]
实施例40化合物53的制备
[0309][0310]
氮气保护下,将化合物1(1g,1.52mmol)、4-溴丁酸2-(三甲基甲硅烷基)乙酯(532mg,2mmol)和k2co3(552mg,4mmol)溶于dmf(15ml)中,升温至40℃搅拌6小时。tlc检测反应完全后,向反应液加入100ml纯水和100ml二氯甲烷进行搅拌。分离有机相,水相继续用二氯甲烷萃取两次(50ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=20/1),得952mg白色泡沫状固体53-1,收率74.2%。
[0311]1h nmr(400mhz,cdcl3)δ8.52(s,1h),8.25(s,1h),5.99(s,1h),5.39(d,j=12.0hz,2h),5.20(d,j=11.3hz,1h),5.10(d,j=11.2hz,1h),4.19

4.14(m,2h),3.88(t,j=5.6hz,2h),2.80(ddd,j=14.4,7.1,3.6hz,2h),2.52

2.37(m,5h),2.13

1.97(m,5h),1.95

1.86(m,2h),1.78(t,j=14.3hz,4h),1.56

1.37(m,12h),1.10

0.94(m,12h),0.72(d,j=7.1hz,3h),0.61(d,j=7.1hz,3h),0.03(s,9h).
[0312]
13
c nmr(101mhz,cdcl3)δ173.33,157.09,156.23,154.61,106.83,105.05,105.01,104.75,93.17,92.10,91.97,81.49,81.41,71.60,71.30,66.79,62.79,51.73,46.11,45.99,37.68,37.65,36.16,34.41,34.37,31.60,31.36,30.85,26.14,26.11,25.06,25.03,24.93,21.44,21.05,20.40,20.36,17.49,13.57,-1.36.
[0313]
ms(esi):[m+h]
+
=845.4
[0314]
氮气保护下,将化合物53-1(845mg,1mmol)溶于四氢呋喃(15ml),降温至0℃后加入tbaf(2.61g,10mmol),加完后慢慢升温至20℃并搅拌2小时。tlc检测反应完全后,反应液减压浓缩、残余物中加入纯水(100ml)和二氯甲烷(100ml),搅拌10分钟后进行分液,水相用二氯甲烷萃取两次(50ml
×
2)。合并有机相,依次用10%柠檬酸水溶液、饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=40/1),得370mg白色固体53,收率49.6%。
[0315]1h nmr(400mhz,cdcl3)δ8.54(s,1h),8.23(s,1h),6.01(s,1h),5.41(d,j=4.1hz,2h),5.21(d,j=11.3hz,1h),5.10(d,j=11.2hz,1h),3.92(dd,j=9.9,5.8hz,2h),2.80(dt,j=10.5,7.3hz,2h),2.59

2.34(m,4h),2.16

1.99(m,4h),1.95

1.86(m,2h),1.84

1.70(m,4h),1.67

1.14(m,17h),1.12

0.89(m,8h),0.72(d,j=7.1hz,3h),0.61(d,j=7.1hz,3h).
[0316]
13
c nmr(101mhz,cdcl3)δ177.95,156.89,155.97,154.44,106.74,104.90,104.85,104.66,93.04,91.89,91.78,81.33,81.24,,71.42,71.10,66.49,51.53,45.91,45.78,37.50,37.43,35.96,34.22,34.15,31.41,30.68,30.58,25.95,25.92,24.84,24.49,21.25,20.84,20.21,20.17,13.38.
[0317]
ms(esi):[m+h]
+
=745.4
[0318]
实施例41化合物54的制备
[0319][0320]
氮气保护下,将化合物1(1g,1.52mmol)、5-溴戊酸2-(三甲基甲硅烷基)乙酯(560mg,2.0mmol)、k2co3(552mg,4mmol)和dmf(15ml)的混合物升温至40℃搅拌6小时。tlc检测反应完全后,向反应液加入100ml纯水和100ml二氯甲烷进行搅拌。分离有机相,水相用二氯甲烷萃取两次(50ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=20/1),得918mg白色泡沫状固体54-1,收率79.6%。
[0321]1h nmr(400mhz,cdcl3)δ8.53(s,1h),8.25(s,1h),6.00(s,1h),5.40(d,j=10.1hz,2h),5.20(d,j=11.3hz,1h),5.12(d,j=11.2hz,1h),4.22

4.08(m,2h),3.93

3.81(m,2h),3.47(s,1h),2.80(ddd,j=12.8,8.7,3.4hz,2h),2.48

2.29(m,4h),2.03(d,j=14.4hz,2h),1.94

1.86(m,2h),1.77(ddd,j=10.8,9.6,6.9hz,8h),1.67

1.49(m,8h),1.45

1.32(m,9h),0.98(td,j=6.7,3.2hz,8h),0.72(d,j=7.2hz,3h),0.61(d,j=7.2hz,3h),0.04(s,9h).
[0322]
13
c nmr(101mhz,cdcl3)δ173.94,157.38,156.74,154.92,107.01,105.37,105.34,105.08,93.45,92.43,92.29,81.83,81.74,71.94,71.65,67.55,62.95,52.05,46.44,46.34,38.01,37.96,36.48,34.73,34.51,31.94,31.18,29.17,26.47,26.43,25.39,25.36,22.21,21.77,21.37,20.72,20.69,17.86,13.89,13.87,-1.05.
[0323]
ms(esi):[m+h]
+
=859.5
[0324]
氮气保护下,将化合物54-1(600mg,0.69mmol)溶于四氢呋喃(15ml),降温至0℃后加入tbaf(1.8g,6.9mmol),加完后慢慢升温至20℃并搅拌2小时。tlc检测反应完全后,反应液减压浓缩、残余物中加入100ml纯水和100ml二氯甲烷进行搅拌。分离有机相,水相用二氯甲烷萃取两次(50ml
×
2)。合并有机相,依次用10%柠檬酸水溶液、饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=40/1),得293mg白色固体54,收率54.9%。
[0325]1h nmr(400mhz,cdcl3)δ8.54(s,1h),8.25(s,1h),6.01(s,1h),5.40(d,j=11.0hz,2h),5.21(d,j=11.3hz,1h),5.12(d,j=11.2hz,1h),3.88(d,j=2.5hz,2h),2.80(ddd,j=14.4,7.2,3.6hz,2h),2.50

2.37(m,4h),2.04(d,j=14.3hz,2h),1.94

1.87(m,2h),1.85

1.69(m,8h),1.62

1.28(m,16h),1.10

0.93(m,8h),0.72(d,j=7.1hz,3h),0.61(d,j=7.1hz,3h).
[0326]
13
c nmr(101mhz,cdcl3)δ177.52,156.92,156.24,154.48,106.63,104.93,104.90,104.67,93.03,91.98,91.84,81.38,81.30,71.49,71.19,67.05,51.60,45.98,45.87,37.56,37.49,36.03,34.28,34.22,33.39,31.49,30.71,28.66,26.02,25.98,24.90,21.58,21.32,20.91,20.24,13.44.
[0327]
ms(esi):[m+h]
+
=759.4
[0328]
实施例42化合物55的制备
[0329][0330]
氮气保护下,将化合物1(1g,1.52mmol)、6-溴己酸2-(三甲基甲硅烷基)乙酯(588mg,2.0mmol)、k2co3(552mg,4mmol)和dmf(15ml)的混合物升温至40℃搅拌6小时。tlc检测反应完全后,向反应液加入100ml纯水和二氯甲烷100ml进行搅拌。分离有机相,水相用二氯甲烷萃取两次(50ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:石油醚/乙酸乙酯=20/1),得1.08g白色泡沫状固体55-1,收率81.7%。
[0331]1h nmr(400mhz,cdcl3)δ8.52(s,1h),8.24(s,1h),5.97(s,1h),5.38(d,j=9.1hz,2h),5.17(d,j=11.3hz,1h),5.10(d,j=11.2hz,1h),4.18

4.08(m,2h),3.87

3.77(m,2h),2.82

2.69(m,2h),2.45

2.22(m,10h),1.92

1.84(m,2h),1.81

1.68(m,6h),1.58

1.31(m,16h),1.10

0.92(m,10h),0.69(d,j=7.2hz,3h),0.58(d,j=7.2hz,3h),0.01(s,9h).
[0332]
13
c nmr(101mhz,cdcl3)δ174.27,157.20,156.80,154.78,106.84,105.38,105.05,93.39,92.39,92.25,81.80,81.74,71.92,71.63,67.85,62.94,51.98,50.97,50.96,46.36,46.26,37.94,37.88,36.41,34.87,34.65,34.61,31.89,31.15,29.37,26.36,26.31,26.26,25.29,25.12,21.67,21.32,20.64,20.61,17.73,13.80,13.78,-1.12.
[0333]
ms(esi):[m+h]
+
=873.5
[0334]
氮气保护下,将化合物55-1(600mg,0.68mmol)溶于四氢呋喃(15ml),降温至0℃后加入tbaf(1.8g,6.9mmol),加完后慢慢升温至20℃并搅拌2小时。tlc检测反应完全后,反应液减压浓缩,向残余物中加入100ml纯水和100ml二氯甲烷进行搅拌。分离有机相,水相继续用二氯甲烷萃取两次(50ml
×
2)。合并有机相,依次用10%柠檬酸水溶液、饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=40/1),得293mg白色固体55,收率55.3%。
[0335]1h nmr(400mhz,cdcl3)δ8.53(s,1h),8.23(s,1h),6.01(s,1h),5.42(s,2h),5.21(d,j=11.3hz,1h),5.12(d,j=11.3hz,1h),3.95

3.79(m,2h),3.48(q,j=7.0hz,1h),2.89

2.71(m,2h),2.52

2.31(m,4h),2.04(d,j=14.5hz,2h),1.97

1.85(m,2h),1.84

1.73(m,6h),1.69(dd,j=15.2,7.6hz,3h),1.63

1.46(m,8h),1.44(d,j=2.1hz,6h),1.36

1.24(m,3h),1.21(t,j=7.0hz,2h),1.05(dd,j=20.6,10.6hz,2h),0.98(dd,j=6.1,1.4hz,6h),0.73(d,j=7.1hz,3h),0.61(d,j=7.1hz,3h).
[0336]
13
c nmr(101mhz,cdcl3)δ177.67,157.07,156.48,154.60,106.67,105.11,105.05,104.82,93.24,92.10,92.00,81.53,81.45,71.64,71.36,67.50,65.99,51.76,46.14,46.02,37.71,37.63,36.18,34.43,34.37,33.84,31.63,30.84,29.03,26.17,
26.14,25.88,25.05,24.57,21.47,21.08,20.42,20.38,15.40,13.56.
[0337]
ms(esi):[m+h]
+
=773.4
[0338]
实施例43化合物56的制备
[0339][0340]
氮气保护下,将化合物1(790mg,1.2mmol)、boc-l-缬氨酸(313mg,1.44mmol)、edci(690mg,3.6mmol)和dmap(73mg,0.6mmol)溶于二氯甲烷(80ml),室温搅拌4小时。tlc检测反应完全后,反应液用饱和碳酸氢钠水溶液洗涤。分离有机相,水相用二氯甲烷萃取两次(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得760mg白色固体56-1,收率为73.9%。
[0341]1h nmr(400mhz,cdcl3)δ8.65(s,1h),8.39(s,1h),6.14(s,1h),5.42(s,1h),5.38(s,1h),5.26(d,j=11.4hz,1h),5.02(d,j=8.7hz,1h),4.79(d,j=11.2hz,1h),4.25(dd,j=8.7,5.3hz,1h),2.81(dd,j=10.8,7.1hz,2h),2.50

2.35(m,2h),2.23(dq,j=13.5,6.7hz,1h),2.10

1.98(m,2h),1.96

1.85(m,2h),1.78(dd,j=20.8,9.2hz,5h),1.66

1.55(m,5h),1.55

1.34(m,19h),1.00(ddd,j=15.9,9.3,5.1hz,14h),0.73(d,j=7.1hz,3h),0.63(d,j=7.1hz,3h).
[0342]
ms(esi):[m+h]
+
=858.5
[0343]
氮气保护下,将化合物56-1(684mg,0.798mmol)溶于二氯甲烷(30ml),在0℃下滴加tfa(2ml,26.9mmol),滴毕,在0℃搅拌2小时。tlc检测反应完全后,向反应液中滴加过量饱和碳酸氢钠水溶液。分离有机相,水相用二氯甲烷萃取两次(30ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1),得425mg的白色固体56,收率71.1%。
[0344]1h nmr(400mhz,cdcl3)δ8.62(s,1h),8.31(s,1h),6.18(s,1h),5.41(s,1h),5.34(s,1h),5.24(d,j=11.4hz,1h),4.71(d,j=11.3hz,1h),3.44(d,j=4.8hz,1h),2.89

2.73(m,2h),2.41(tt,j=14.1,3.9hz,2h),2.23

2.09(m,1h),2.08

1.97(m,2h),1.95

1.84(m,2h),1.82

1.68(m,4h),1.60

1.31(m,16h),1.06(d,j=6.8hz,4h),0.97(t,j=5.9hz,9h),0.72(d,j=7.1hz,3h),0.61(d,j=7.2hz,3h).
[0345]
13
c nmr(101mhz,cdcl3)δ173.19,156.91,154.49,148.23,111.96,108.52,105.30,105.18,102.80,92.04,92.00,81.45,81.42,72.27,71.57,60.62,60.43,51.74,51.69,46.08,45.83,37.74,37.68,36.18,36.14,34.43,34.31,32.33,31.35,30.65,26.20,26.18,25.13,25.05,21.50,21.29,21.12,20.49,20.43,19.79,17.36,14.44,13.92,13.61.
[0346]
ms(esi):[m+h]
+
=758.4
[0347]
实施例44化合物57的制备
[0348][0349]
氮气保护下,将化合物1(1317mg,2.0mmol)、boc-o-bn-l-苏氨酸(742mg,2.4mmol)、edci(1150mg,6.0mmol)和dmap(122mg,1.0mmol)溶于二氯甲烷(120ml),室温搅拌15小时。tlc检测反应完全后,反应液用饱和碳酸氢钠溶液洗涤,水相用二氯甲烷萃取两次(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=120/1),得1.5g白色固体57-1,收率为79.0%。
[0350]1h nmr(400mhz,cdcl3)δ8.63(d,j=5.2hz,1h),8.33(d,j=34.7hz,1h),7.32(dt,j=18.5,4.5hz,5h),6.16(d,j=21.8hz,1h),5.42(s,1h),5.34(d,j=9.3hz,1h),5.26(d,j=11.3hz,1h),4.80

4.68(m,1h),4.62

4.46(m,2h),4.38(dd,j=9.3,2.4hz,1h),4.32

4.20(m,1h),2.90

2.68(m,2h),2.42(qd,j=14.7,3.8hz,2h),2.04(dd,j=12.3,5.0hz,2h),1.95

1.73(m,4h),1.68

1.51(m,8h),1.48

1.40(m,16h),1.37

1.28(m,7h),0.95(dd,j=16.5,5.9hz,8h),0.73(d,j=7.1hz,3h),0.59(d,j=7.1hz,3h).
[0351]
ms(esi):[m+h]
+
=950.5
[0352]
氮气保护下,将化合物57-1(554mg,0.583mmol)溶于二氯甲烷(24ml),在0℃下滴加tfa(1ml,13.5mmol),滴毕,在0℃搅拌2小时。tlc检测反应完全后,向反应液中滴加过量饱和碳酸氢钠水溶液。分离有机相,水相用二氯甲烷萃取两次(30ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1),得406mg白色固体57,收率81.9%。
[0353]1h nmr(400mhz,cdcl3)δ8.63(s,1h),8.35(s,1h),7.36

7.27(m,5h),6.20(s,1h),5.42(s,1h),5.26(d,j=11.3hz,1h),5.12(s,1h),4.71(d,j=11.2hz,1h),4.63

4.53(m,2h),4.06(dt,j=10.3,5.1hz,1h),3.62(d,j=3.9hz,1h),2.86

2.76(m,2h),2.48

2.35(m,2h),2.08

1.99(m,2h),1.94

1.70(m,8h),1.53

1.26(m,16h),1.13

0.95(m,6h),0.91(d,j=6.1hz,3h),0.73(d,j=7.1hz,3h),0.59(d,j=7.1hz,3h).
[0354]
13
c nmr(101mhz,cdcl3)δ171.77,156.86,154.42,148.17,138.29,128.60,127.85,127.67,111.97,108.44,105.12,105.10,102.78,91.91,91.79,81.36,81.30,72.20,71.48,71.42,59.85,51.65,51.50,45.99,45.69,37.65,37.28,36.09,36.04,34.33,34.12,31.26,30.60,26.10,25.04,24.91,21.40,20.90,20.40,20.27,16.60,13.88,13.51.
[0355]
ms(esi):[m+h]
+
=850.4
[0356]
实施例45化合物58的制备
[0357][0358]
氮气保护下,将化合物1(987.5mg,1.5mmol)、boc-l-亮氨酸(416mg,1.8mmol),edci(863mg,4.5mmol)和dmap(92mg,0.75mmol)溶于二氯甲烷(80ml),室温搅拌4小时。tlc检测反应完全后,反应液用饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取两次(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=200/1),得1.08g白色固体58-1,收率为82.6%。
[0359]1h nmr(400mhz,cdcl3)δ8.64(s,1h),8.38(s,1h),6.14(s,1h),5.40(d,j=11.6hz,2h),5.26(d,j=11.4hz,1h),4.87(dd,j=26.6,9.6hz,2h),4.37(dd,j=14.1,8.5hz,1h),2.82(dd,j=10.9,7.2hz,2h),2.48

2.35(m,2h),2.08

1.99(m,2h),1.95

1.86(m,2h),1.78(ddd,j=19.4,12.4,6.2hz,7h),1.67

1.55(m,7h),1.48

1.39(m,17h),1.33(dt,j=12.9,5.6hz,3h),1.01

0.94(m,12h),0.73(d,j=7.1hz,3h),0.63(d,j=7.1hz,3h).
[0360]
ms(esi):[m+h]
+
=872.5
[0361]
氮气保护下,将化合物58-1(600mg,0.688mmol)溶于二氯甲烷(20ml),在0℃下滴加tfa(1ml,13.5mmol),滴毕,在0℃搅拌2.5小时。tlc检测反应完全后,向反应液中滴加过量饱和碳酸氢钠水溶液。分离有机相,水相用二氯甲烷萃取两次(30ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1),得420mg白色固体58,收率79.3%。
[0362]1h nmr(400mhz,cdcl3)δ8.62(s,1h),8.29(s,1h),6.21(s,1h),5.42(s,1h),5.36(s,1h),5.26(d,j=11.4hz,1h),4.75(d,j=11.3hz,1h),3.65(dd,j=9.1,4.9hz,1h),2.82(dt,j=7.9,5.5hz,2h),2.49

2.37(m,2h),2.04(d,j=14.1hz,2h),1.90(dd,j=7.4,4.8hz,4h),1.82

1.68(m,7h),1.63

1.49(m,6h),1.41(dd,j=16.3,4.1hz,9h),1.01

0.93(m,13h),0.73(d,j=7.1hz,3h),0.60(d,j=7.1hz,3h).
[0363]
13
c nmr(101mhz,cdcl3)δ174.55,157.34,154.87,148.53,112.40,108.84,105.74,105.62,103.36,92.45,92.43,81.87,81.81,72.58,71.99,53.67,52.15,52.08,46.48,46.23,44.83,38.17,38.13,36.59,36.55,34.84,34.63,31.74,31.02,26.60,25.54,25.45,23.82,22.26,21.91,21.75,20.90,20.84,14.36,14.02.
[0364]
ms(esi):[m+h]
+
=772.4
[0365]
实施例46化合物59的制备
[0366][0367]
氮气保护下,将化合物1(790mg,1.2mmol)、boc-l-异亮氨酸(333mg,1.44mmol)、edci(690mg,3.6mmol)和dmap(73mg,0.6mmol)溶于二氯甲烷(80ml)后,室温搅拌4小时。tlc检测反应完全后,反应液用饱和碳酸氢钠水溶液洗涤。分离有机相,水相用二氯甲烷萃取两次(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得560mg白色固体59-1,收率53.5%。
[0368]1h nmr(400mhz,cdcl3)δ8.66(s,1h),8.40(s,1h),6.13(s,1h),5.42(s,1h),5.38(s,1h),5.26(d,j=11.4hz,1h),5.03(d,j=8.6hz,1h),4.79(d,j=11.2hz,1h),4.31(dd,j=8.5,5.2hz,1h),2.81(dd,j=10.9,7.1hz,2h),2.49

2.34(m,2h),2.04(d,j=13.6hz,2h),1.97

1.85(m,3h),1.79(dd,j=20.2,8.3hz,5h),1.66

1.55(m,7h),1.50

1.40(m,17h),1.30(ddd,j=15.4,12.3,6.5hz,4h),1.05(d,j=6.8hz,4h),0.97(dd,j=6.2,3.7hz,8h),0.73(d,j=7.1hz,3h),0.64(d,j=7.1hz,3h).
[0369]
ms(esi):[m+h]
+
=872.5
[0370]
氮气保护下,将化合物59-1(468mg,0.537mmol)溶于二氯甲烷(20ml),在0℃下向其中滴加znbr2的二氯甲烷溶液(5ml,468mg,2.078mmol),滴毕,在0℃继续搅拌3小时。tlc检测反应完全后,过滤反应液除去不溶性固体。滤液减压浓缩,得淡黄色泡沫状固体340mg,用10ml二氯甲烷和1ml甲醇的混合溶剂体系重结晶,过滤,收集滤饼,50℃真空干燥得244mg白色固体产品59,收率71.7%。
[0371]1h nmr(400mhz,cdcl3)δ8.64(s,1h),8.32(s,1h),6.18(s,1h),5.42(s,1h),5.35(s,1h),5.26(d,j=11.4hz,1h),4.71(d,j=11.3hz,1h),3.51(d,j=4.9hz,1h),2.90

2.74(m,2h),2.43(tt,j=14.1,4.1hz,2h),2.08

2.00(m,2h),1.90(ddd,j=16.7,8.7,5.2hz,3h),1.84

1.72(m,4h),1.63

1.37(m,18h),1.05(t,j=6.5hz,5h),0.95(dd,j=13.7,6.3hz,9h),0.73(d,j=7.1hz,3h),0.63(d,j=7.1hz,3h).
[0372]
13
c nmr(101mhz,cdcl3)δ173.19,156.91,154.49,148.23,111.96,108.52,105.30,105.18,102.80,92.04,92.00,81.45,81.42,72.27,71.57,60.62,60.43,51.74,51.69,46.08,45.83,37.74,37.68,36.18,36.14,34.43,34.31,32.33,31.35,30.65,26.20,26.18,25.13,25.05,21.50,21.29,21.12,20.49,20.43,19.79,17.36,14.44,13.92,13.61.
[0373]
ms(esi):[m+h]
+
=772.4
[0374]
实施例47化合物60的制备
[0375][0376]
氮气保护下,将化合物1(790mg,1.2mmol)、boc-l-叔亮氨酸(313mg,1.44mmol)、edci(690mg,3.6mmol)和dmap(73mg,0.6mmol)溶于二氯甲烷(80ml)后,室温搅拌24小时。tlc检测反应完全后,反应液用饱和碳酸氢钠水溶液洗涤,分离有机相,水相用二氯甲烷萃取两次(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得670mg白色固体60-1,收率为64.1%。
[0377]1h nmr(400mhz,cdcl3)δ8.65(s,1h),8.42(s,1h),6.14(s,1h),5.42(s,1h),5.37(s,1h),5.26(d,j=11.3hz,1h),5.09(d,j=9.1hz,1h),4.83(d,j=11.2hz,1h),4.15(d,j=9.2hz,1h),2.88

2.73(m,2h),2.49

2.37(m,2h),2.04(d,j=13.3hz,2h),1.95

1.87(m,2h),1.78(dd,j=21.6,7.2hz,5h),1.59(s,5h),1.48

1.40(m,16h),1.33(dd,j=10.9,6.9hz,3h),1.08(s,11h),0.96(t,j=6.0hz,6h),0.73(d,j=7.1hz,3h),0.66(d,j=7.1hz,3h).
[0378]
ms(esi):[m+h]
+
:872.5
[0379]
氮气保护下,将化合物60-1(504mg,0.578mmol)溶解于二氯甲烷(24ml),降温至0℃后向其中滴加tfa(1ml,13.5mmol),滴毕,在0℃继续搅拌3小时。tlc检测反应完全后,向反应液中滴加过量饱和碳酸氢钠水溶液。分离有机相,水相用二氯甲烷萃取两次(30ml
×
2),合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得312mg白色固体60,收率69.9%。
[0380]1h nmr(400mhz,cdcl3)δ8.63(s,1h),8.33(s,1h),6.21(s,1h),5.42(s,1h),5.36(s,1h),5.26(d,j=11.4hz,1h),4.75(d,j=11.3hz,1h),3.29(s,1h),2.82(dtd,j=11.0,7.4,3.8hz,2h),2.42(tt,j=13.9,3.9hz,2h),2.04(d,j=13.6hz,2h),1.96

1.85(m,2h),1.78(dd,j=21.2,8.8hz,4h),1.66

1.22(m,16h),1.14

1.00(m,11h),0.97(d,j=6.0hz,6h),0.73(d,j=7.1hz,3h),0.63(d,j=7.1hz,3h).
[0381]
13
c nmr(101mhz,cdcl3)δ172.43,158.67,156.74,156.17,154.45,149.89,147.98,122.34,111.87,109.41,108.49,108.04,105.21,105.10,103.54,102.68,99.31,91.96,91.90,91.44,81.36,72.22,71.48,64.00,51.65,51.61,45.99,45.76,37.66,37.60,36.09,36.06,34.98,34.34,34.22,31.26,30.61,26.58,26.11,26.06,25.04,24.96,21.41,21.09,20.40,20.34,13.82,13.51.
[0382]
ms(esi):[m+h]
+
=772.4
[0383]
实施例48化合物61的制备
[0384][0385]
氮气保护下,将化合物1(790mg,1.2mmol)、boc-4-氨基苯甲酸(342mg,1.44mmol)、edci(690mg,3.6mmol)和dmap(73mg,0.6mmol)溶于二氯甲烷(80ml)后,室温搅拌7小时。tlc检测反应完全后,反应液用饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取两次(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得770mg白色固体61-1,收率为71.8%。
[0386]1h nmr(400mhz,cdcl3)δ8.62(s,1h),8.34(s,1h),8.08(d,j=8.8hz,2h),7.49(d,j=8.8hz,2h),6.75(s,1h),6.33(s,1h),5.44(s,1h),5.32(s,1h),5.29(d,j=11.3hz,1h),4.80(d,j=11.3hz,1h),2.84(ddd,j=11.5,7.4,4.4hz,2h),2.43(qd,j=14.3,3.9hz,2h),2.04(d,j=14.5hz,2h),1.98

1.85(m,2h),1.80(dd,j=20.3,9.1hz,2h),1.70

1.58(m,8h),1.57

1.32(m,19h),0.95(dd,j=17.5,5.9hz,8h),0.76(d,j=7.1hz,3h),0.65(d,j=7.1hz,3h).
[0387]
ms(esi):[m+h]
+
=878.4
[0388]
氮气保护下,将化合物61-1(470mg,0.526mmol)溶解于二氯甲烷(20ml),降温至0℃后向其中滴加tfa(1ml,13.5mmol),滴毕,0℃继续搅拌3小时。tlc检测反应完全后,向反应液滴加过量饱和碳酸氢钠水溶液。分离有机相,水相用二氯甲烷萃取两次(30ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1),得317mg白色固体61,收率76.6%。
[0389]1h nmr(400mhz,cdcl3)δ8.59(s,1h),8.33(s,1h),7.94(d,j=8.6hz,2h),6.68(d,j=8.6hz,2h),6.31(s,1h),5.43(s,1h),5.32(s,1h),5.30(s,2h),4.82(d,j=11.2hz,1h),4.18(s,2h),2.92

2.78(m,2h),2.49

2.35(m,2h),2.04(d,j=14.5hz,2h),1.95

1.74(m,4h),1.66

1.30(m,18h),1.11

0.91(m,7h),0.75(d,j=7.1hz,3h),0.66(d,j=7.1hz,3h).
[0390]
13
c nmr(101mhz,cdcl3)δ164.15,156.80,154.28,151.55,148.62,132.33,118.87,114.07,111.55,108.81,105.16,105.10,103.58,91.94,81.38,81.37,72.40,71.54,53.58,51.68,51.59,46.02,45.76,37.67,37.50,36.11,36.07,34.37,34.18,31.25,30.64,26.12,25.05,24.96,21.43,20.81,20.41,20.31,14.00,13.55.
[0391]
ms(esi):[m+h]
+
:778.4
[0392]
实施例49化合物62的制备
[0393][0394]
氮气保护下,将化合物1(790mg,1.2mmol)、boc-4-氨基环己酸(350mg,1.44mmol)、edci(690mg,3.6mmol)和dmap(73mg,0.6mmol)溶于二氯甲烷(80ml)后,室温搅拌6小时。tlc检测反应完全后,反应液用饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取两次(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得720mg白色固体62-1,收率为67.9%。
[0395]1h nmr(400mhz,cdcl3)δ8.62(d,j=6.8hz,1h),8.31(d,j=3.6hz,1h),6.18(d,j=9.6hz,1h),5.44(s,1h),5.38(s,1h),5.28(d,j=11.4hz,1h),4.70(d,j=11.5hz,1h),4.45(s,1h),3.47(s,1h),2.92

2.78(m,2h),2.52

2.40(m,3h),2.15(s,3h),2.11

2.04(m,2h),1.98

1.89(m,2h),1.80(s,4h),1.62(d,j=14.5hz,10h),1.50

1.33(m,19h),1.23

1.07(m,3h),1.00(t,j=5.7hz,6h),0.75(d,j=7.1hz,3h),0.61(d,j=7.3hz,3h).
[0396]
ms(esi):[m+h]
+
=884.5
[0397]
氮气保护下,将化合物62-1(555mg,0.628mmol)溶解于二氯甲烷(33ml),降温至0℃后向其中滴加tfa(1ml,13.5mmol),滴毕,0℃继续搅拌3小时。tlc检测反应完全后,向反应液中滴加过量饱和碳酸氢钠水溶液。分离有机相,水相用二氯甲烷萃取两次(30ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1),得257mg白色固体62,收率52.3%。
[0398]1h nmr(400mhz,cdcl3)δ8.71(s,1h),8.44(s,1h),6.14(s,1h),5.39(s,1h),5.31(s,1h),5.21(d,j=11.2hz,1h),4.66(d,j=11.1hz,1h),3.08(d,j=7.3hz,1h),2.77(d,j=7.2hz,2h),2.45(d,j=39.5hz,3h),2.19(s,4h),2.02(d,j=9.9hz,2h),1.94

1.69(m,6h),1.64

1.10(m,22h),0.94(dd,j=11.6,5.5hz,8h),0.70(d,j=6.8hz,3h),0.57(d,j=6.8hz,3h).
[0399]
13
c nmr(101mhz,cdcl3)δ173.19,156.91,154.49,148.23,111.96,108.52,105.30,105.18,102.80,92.04,92.00,81.45,81.42,72.27,71.57,60.62,60.43,51.74,51.69,46.08,45.83,37.74,37.68,36.18,36.14,34.43,34.31,32.33,31.35,30.65,26.20,26.18,25.13,25.05,21.50,21.29,21.12,20.49,20.43,19.79,17.36,14.44,13.92,13.61.
[0400]
ms(esi):[m+h]
+
=784.4
[0401]
实施例50化合物63的制备
[0402]
[0403]
氮气保护下,将化合物1(790mg,1.2mmol)、boc-l-新戊基氨基酸(353mg,1.44mmol)、edci(690mg,3.6mmol)和dmap(73mg,0.6mmol)溶于二氯甲烷(80ml)后,室温搅拌4小时。tlc检测反应完全后,反应液用饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取两次(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得860mg白色固体63-1,收率为80.9%。
[0404]1h nmr(400mhz,cdcl3)δ8.63(s,1h),8.38(s,1h),6.14(s,1h),5.40(d,j=10.6hz,2h),5.26(d,j=11.3hz,1h),4.84(t,j=10.1hz,2h),4.40(td,j=8.9,2.9hz,1h),2.81(ddd,j=10.8,7.2,3.7hz,2h),2.51

2.32(m,2h),2.10

1.98(m,2h),1.97

1.87(m,2h),1.82(ddd,j=32.4,18.7,7.0hz,6h),1.68

1.55(m,6h),1.43(d,j=4.7hz,17h),1.31(ddd,j=17.2,11.1,6.3hz,3h),1.02(s,10h),0.97(dd,j=5.9,3.0hz,6h),0.73(d,j=7.1hz,3h),0.63(d,j=7.1hz,3h).
[0405]
ms(esi):[m+h]
+
=886.5
[0406]
氮气保护下,将化合物63-1(510mg,0.576mmol)溶解于二氯甲烷(23ml),降温至0℃后向其中滴加tfa(1ml,13.5mmol),滴毕,在0℃继续搅拌2小时。tlc检测反应完全后,向反应液中滴加过量饱和碳酸氢钠水溶液。分离有机相,水相用二氯甲烷萃取两次(30ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=50/1),得350mg白色固体63,收率77.4%。
[0407]1h nmr(400mhz,cdcl3)δ8.61(s,1h),8.29(s,1h),6.20(s,1h),5.42(s,1h),5.36(s,1h),5.26(d,j=11.3hz,1h),4.77(d,j=11.3hz,1h),3.65(dd,j=7.4,4.1hz,1h),2.89

2.76(m,2h),2.50

2.37(m,2h),2.04(dd,j=14.4,2.7hz,2h),1.97

1.87(m,3h),1.83

1.71(m,4h),1.69

1.59(m,3h),1.57

1.26(m,16h),1.02(s,9h),0.99

0.94(m,6h),0.73(d,j=7.1hz,3h),0.61(d,j=7.2hz,3h).
[0408]
13
c nmr(101mhz,cdcl3)δ174.29,156.85,154.35,148.11,111.87,108.37,105.24,105.11,102.77,91.97,91.93,81.37,81.31,72.06,71.48,52.71,51.66,51.58,48.99,45.98,45.73,37.67,37.64,36.09,36.05,34.34,34.13,31.24,30.93,30.53,30.04,26.10,25.04,24.96,21.41,21.28,20.40,20.34,13.83,13.52.
[0409]
ms(esi):[m+h]
+
=786.4
[0410]
实施例51化合物64的制备
[0411][0412]
氮气保护下,将化合物1(790mg,1.2mmol)、boc-4-氨基苯基乙酸(362mg,1.44mmol)、edci(690mg,3.6mmol)和dmap(73mg,0.6mmol)溶于二氯甲烷(80ml)后,室温搅拌4小时。tlc检测反应完全后,反应液用饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取两
次(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得730mg白色固体64-1,收率为68.2%。
[0413]1h nmr(400mhz,cdcl3)δ8.60(s,1h),8.20(s,1h),7.37(d,j=8.4hz,2h),7.30(d,j=8.5hz,2h),6.50(s,1h),6.20(s,1h),5.41(s,1h),5.23(d,j=11.4hz,1h),5.10(s,1h),4.31(d,j=11.3hz,1h),3.73(q,j=14.0hz,2h),2.85

2.63(m,2h),2.49

2.32(m,2h),2.08

1.96(m,2h),1.94

1.83(m,2h),1.81

1.70(m,2h),1.61(s,6h),1.51(s,11h),1.42(d,j=6.1hz,8h),1.34

1.23(m,4h),0.96(t,j=5.5hz,6h),0.70(d,j=7.1hz,3h),0.39(d,j=7.1hz,3h).
[0414]
ms(esi):[m+h]
+
=892.4
[0415]
氮气保护下,将化合物64-1(570mg,0.639mmol)溶解于二氯甲烷(26ml),降温至0℃后向其中滴加tfa(1ml,13.5mmol),滴毕,在0℃继续搅拌3小时。tlc检测反应完全后,滴加过量饱和碳酸氢钠水溶液。收集有机相,水相用二氯甲烷萃取两次(30ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=100/1),得334mg白色固体64,收率66.1%。
[0416]1h nmr(400mhz,cdcl3)δ8.60(s,1h),8.21(s,1h),7.14(d,j=7.3hz,2h),6.63(d,j=7.3hz,2h),6.19(s,1h),5.40(s,1h),5.23(d,j=11.1hz,1h),5.14(s,1h),4.37(d,j=10.9hz,1h),3.65(q,j=13.8hz,4h),2.74(d,j=43.4hz,2h),2.40(dd,j=26.6,13.2hz,2h),2.02(d,j=12.4hz,2h),1.88(s,2h),1.81

1.67(m,3h),1.65

1.17(m,18h),0.96(s,7h),0.70(d,j=6.2hz,3h),0.40(d,j=6.2hz,3h).
[0417]
13
c nmr(101mhz,cdcl3)δ168.95,156.41,153.93,147.82,145.61,130.00,122.85,114.92,111.44,108.33,104.78,104.74,102.79,91.56,81.01,71.50,71.15,51.30,51.17,45.63,45.20,40.77,37.30,37.18,35.75,35.70,33.98,30.91,29.99,25.76,24.69,24.63,21.04,20.54,20.06,13.28,13.14.
[0418]
ms(esi):[m+h]
+
=792.4
[0419]
实施例52化合物65的制备
[0420][0421]
氮气保护下,将化合物1(790mg,1.2mmol)、boc-4-苯丙氨酸(382mg,1.44mmol)、edci(690mg,3.6mmol)和dmap(73mg,0.6mmol)溶于二氯甲烷(80ml)后,室温搅拌4小时。tlc检测反应完全后,反应液用饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取两次(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得800mg白色固体65-1,收率为81.0%。
[0422]1h nmr(400mhz,cdcl3)δ8.64(s,1h),8.38(s,1h),7.35(t,j=7.2hz,2h),7.29(d,j=7.2hz,1h),7.25

7.20(m,2h),6.04(s,1h),5.42(s,1h),5.34(s,1h),5.25(d,j=
11.4hz,1h),4.97(d,j=8.0hz,1h),4.72(d,j=11.2hz,1h),4.67(dd,j=14.2,6.6hz,1h),3.26

3.09(m,2h),2.87

2.69(m,2h),2.50

2.33(m,2h),2.12

1.97(m,2h),1.90(dd,j=7.1,3.2hz,2h),1.77(ddd,j=33.7,16.7,8.8hz,4h),1.67

1.55(m,7h),1.41(dd,j=18.8,13.0hz,18h),1.32(dd,j=11.7,5.1hz,2h),0.97(dd,j=5.9,3.6hz,6h),0.73(d,j=7.1hz,3h),0.60(d,j=7.1hz,3h).
[0423]
ms(esi):[m+na]
+
=928.4
[0424]
氮气保护下,将化合物65-1(436mg,0.481mmol)溶解于二氯甲烷(26ml),降温至0℃后向其中滴加tfa(1ml,13.5mmol),滴毕,在0℃继续搅拌3小时。tlc检测反应完全后,滴加过量饱和碳酸氢钠水溶液。分离有机相,水相用二氯甲烷萃取两次(30ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=60/1),得230mg白色固体65,收率59.3%。
[0425]1h nmr(400mhz,cdcl3)δ8.64(s,1h),8.33(s,1h),7.38

7.25(m,5h),6.21(s,1h),5.42(s,1h),5.31(s,1h),5.26(d,j=11.3hz,1h),4.67(d,j=11.3hz,1h),3.91(dd,j=7.5,5.3hz,1h),3.25(dd,j=13.5,5.0hz,1h),2.97(dd,j=13.5,7.8hz,1h),2.82(d,j=7.1hz,2h),2.43(qd,j=13.1,3.6hz,2h),2.05(d,j=14.1hz,2h),1.98

1.86(m,2h),1.79(dd,j=19.0,10.6hz,2h),1.72

1.35(m,18h),1.12

0.92(m,8h),0.74(d,j=7.1hz,3h),0.57(d,j=7.1hz,3h).
[0426]
13
c nmr(101mhz,cdcl3)δ172.11,156.65,154.16,147.74,136.84,129.38,128.64,126.95,111.78,108.08,105.01,104.91,102.60,91.72,81.16,81.10,71.93,71.28,51.45,51.35,45.78,45.46,37.46,37.33,35.89,35.84,34.14,33.97,31.03,30.36,25.91,24.84,24.77,21.21,20.73,20.20,20.11,13.66,13.33.
[0427]
ms(esi):[m+h]
+
=806.4
[0428]
实施例53化合物66的制备
[0429][0430]
氮气保护下,将化合物1(1317mg,2.0mmol)、boc-o-bn-l-酪氨酸(762mg,2.40mmol)、edci(1150mg,6.0mmol)和dmap(122mg,1.0mmol)溶于二氯甲烷(120ml)后,室温搅拌4小时。tlc检测反应完全后,反应液用饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取两次(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得1.7g白色固体66-1,收率为84.0%。
[0431]1h nmr(400mhz,cdcl3)δ8.65(s,1h),8.39(s,1h),7.44(d,j=7.1hz,2h),7.38(dd,j=9.9,4.7hz,2h),7.31(dd,j=8.5,5.9hz,1h),7.15(d,j=8.6hz,2h),6.96(d,j=8.5hz,2h),6.10(s,1h),5.42(s,1h),5.25(d,j=11.4hz,1h),5.13

5.00(m,2h),4.94(d,j
=8.0hz,1h),4.64(dd,j=13.8,8.6hz,2h),3.12(d,j=6.0hz,2h),2.89

2.69(m,2h),2.42(ddd,j=21.3,15.8,3.6hz,2h),2.04(d,j=14.9hz,2h),1.95

1.67(m,6h),1.66

1.54(m,7h),1.49

1.36(m,18h),1.32(d,j=6.8hz,3h),0.96(dd,j=11.0,6.1hz,6h),0.73(d,j=7.1hz,3h),0.60(d,j=7.1hz,3h).
[0432]
ms(esi):[m+h]
+
=1012.5
[0433]
氮气保护下,将化合物66-1(416mg,0.411mmol)溶解于二氯甲烷(24ml),降温至0℃后向其中滴加tfa(1ml,13.5mmol),滴毕,在0℃继续搅拌2小时。tlc检测反应完全后,滴加过量饱和碳酸氢钠水溶液。分离有机相,水相用二氯甲烷萃取两次(30ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=60/1),得266mg白色固体66,收率71.1%。
[0434]1h nmr(400mhz,cdcl3)δ8.65(s,1h),8.34(s,1h),7.46

7.30(m,5h),7.18(d,j=8.3hz,2h),6.95(d,j=8.3hz,2h),6.22(s,1h),5.42(s,1h),5.35

5.20(m,2h),5.05(d,j=2.7hz,2h),4.62(d,j=11.2hz,1h),3.88(s,1h),3.16(s,1h),2.97(d,j=6.6hz,1h),2.81(d,j=4.6hz,2h),2.42(dd,j=26.9,12.8hz,2h),2.03(t,j=13.4hz,2h),1.81

1.67(m,6h),1.57

1.24(m,16h),1.14

0.83(m,10h),0.73(d,j=7.0hz,3h),0.57(d,j=7.1hz,3h).
[0435]
13
c nmr(101mhz,cdcl3)δ158.01,156.84,154.37,147.93,137.14,130.70,129.10,128.68,128.05,127.55,115.18,111.97,108.26,105.20,105.12,102.81,91.92,81.37,81.31,72.12,71.49,70.09,51.66,51.55,45.98,45.67,37.67,37.43,36.09,36.05,34.34,34.18,31.24,30.59,26.11,25.04,24.88,21.41,20.93,20.40,20.31,13.86,13.53.
[0436]
ms(esi):[m+h]
+
=912.4
[0437]
实施例54化合物67的制备
[0438][0439]
氮气保护下,将化合物1(790mg,1.2mmol)、boc-l-色氨酸(438mg,1.44mmol)、edci(690mg,3.6mmol)和dmap(73mg,0.6mmol)溶于二氯甲烷(80ml)后,室温搅拌3小时。tlc检测反应完全后,反应液用饱和碳酸氢钠水溶液洗涤,水相用二氯甲烷萃取两次(100ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸钠干燥,减压浓缩,残余物经硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=400/1~200/1),得800mg白色固体67-1,收率为70.4%。
[0440]1h nmr(400mhz,cdcl3)δ8.65(s,1h),8.36(s,1h),8.29(s,1h),7.64(d,j=7.8hz,1h),7.38(d,j=8.0hz,1h),7.21(t,j=7.0hz,1h),7.15(t,j=7.4hz,1h),7.07(s,1h),6.12(s,1h),5.41(s,1h),5.23(d,j=11.4hz,1h),5.15(s,1h),5.07(d,j=8.0hz,1h),4.80(dd,j=13.5,5.8hz,1h),4.56(d,j=11.2hz,1h),3.37(d,j=5.6hz,2h),2.87

2.60(m,2h),2.42(td,j=17.2,3.8hz,2h),2.11

1.97(m,2h),1.97

1.83(m,2h),1.83

1.70(m,
2h),1.69

1.52(m,8h),1.45

1.36(m,16h),1.34

1.19(m,4h),0.95(dd,j=15.4,5.4hz,7h),0.71(d,j=7.1hz,3h),0.53(d,j=7.1hz,3h).
[0441]
ms(esi):[m+h]
+
=945.5
[0442]
氮气保护下,将化合物67-1(445mg,0.471mmol)溶解于二氯甲烷(27ml),降温至0℃后向其中滴加tfa(1ml,13.5mmol),滴毕,在0℃继续搅拌3小时。tlc检测反应完全后,滴加过量饱和碳酸氢钠水溶液,分离有机相,水相用二氯甲烷萃取两次(30ml
×
2)。合并有机相,用饱和食盐水洗涤,无水硫酸镁干燥,减压浓缩,残余物硅胶柱层析纯化(洗脱剂:二氯甲烷/甲醇=50/1),得297mg白色固体67,收率74.6%。
[0443]1h nmr(400mhz,cdcl3)δ8.65(s,1h),8.44(s,1h),8.33(s,1h),7.65(d,j=7.7hz,1h),7.38(d,j=8.0hz,1h),7.19(t,j=7.3hz,1h),7.12(t,j=7.3hz,1h),7.05(s,1h),6.27(s,1h),5.42(s,1h),5.26(d,j=11.3hz,1h),5.06(s,1h),4.55(d,j=11.2hz,1h),4.04(s,1h),3.39(dd,j=14.4,4.5hz,1h),3.18(dd,j=14.2,6.5hz,1h),2.78(d,j=36.7hz,2h),2.50

2.33(m,2h),2.03(s,2h),1.87(d,j=15.1hz,2h),1.74(s,6h),1.53

1.23(m,16h),0.97(d,j=5.6hz,3h),0.90(d,j=5.6hz,3h),0.72(d,j=6.9hz,3h),0.52(d,j=6.9hz,3h).
[0444]
13
c nmr(101mhz,cdcl3)δ172.75,156.79,154.37,147.98,136.30,127.83,123.42,122.31,119.72,118.76,111.91,111.50,110.88,108.35,105.19,105.16,102.96,91.94,91.89,81.39,81.32,72.08,71.49,55.72,51.65,51.45,45.97,45.60,37.66,37.25,36.10,36.05,34.31,34.05,31.24,30.57,26.12,25.04,24.95,21.38,20.68,20.39,20.20,13.77,13.51.
[0445]
ms(esi):[m+h]
+
=845.4
[0446]
实施例55化合物1~67对vero细胞的增殖抑制作用
[0447]
实验目的:验证双-(10-去氧双氢青蒿素)-间苯三酚衍生物样品1~67对vero细胞(非洲绿猴肾细胞)的增殖抑制作用。
[0448]
实验原理:利用cell counting kit-8(cck-8)试剂盒检测肿瘤细胞在不同浓度药物作用下的存活率,graphpad prism 7软件拟合出细胞的生长抑制曲线,取50%抑制率时的横坐标即为半数抑制浓度(ic
50
)。cck-8染料能被细胞内脱氢酶还原生成的橙黄色的甲臜产物,生成的甲臜量与活细胞数量成正比,酶标仪在波长450nm处测得的吸光度可间接反映细胞数目,此方法灵敏度高、重复性好、结果可靠,广泛应用于细胞毒性测试。
[0449]
实验方法:用培养基培养细胞后,贴壁细胞弃去原培养基,pbs洗涤,胰酶消化后收集细胞;悬浮细胞直接收集,离心后弃去上清液,细胞用新鲜培养基打匀后计数,细胞密度调整至4
×
104/ml,接种于透明的96孔板中,每孔接种100μl细胞悬液,即每孔4000个细胞。每块96孔板中设有只加培养基的空白孔,以及只加细胞不加药物的对照孔。加药孔每孔药物的终浓度分别为5.0、1.25、0.313、0.078、0.020、0.005和0.001μm,每组设三复孔。接种后的96孔板在37℃,5%co2培养箱中继续培养72小时,在结束前2小时,每孔加10μl cck-8溶液,置于37℃培养箱中直至培养结束。全自动酶标仪于450nm波长处测定o.d值,graphpad prism 7拟合出细胞生长抑制曲线,取50%抑制率时的横坐标值,即为药物半数抑制浓度(ic
50
)。
[0450]
计算公式如下:
[0451]
细胞存活率(%)=(化合物加药孔的o.d值﹣空白孔的o.d值)/(对照孔的o.d值﹣空白孔的o.d值)
×
100%
[0452]
药物抑制率(%)=[1﹣(化合物加药孔的o.d值﹣空白孔的o.d值)/(对照孔的o.d值﹣空白孔的o.d值)]
×
100%
[0453]
实验结果:ic
50
结果见表1
[0454]
表1 双-(10-去氧双氢青蒿素)-间苯三酚衍生物对vero细胞的增殖抑制活性ic
50
(μm)
[0455]
[0456]
[0457][0458]
从表1的数据可以发现,化合物19~67对vero细胞的增殖抑制活性ic
50
数值至少是化合物1~18的10倍以上,说明化合物19~67的细胞毒性远低于化合物1~18。
[0459]
实施例56化合物1及化合物14~67对pha刺激的人pbmc体外增殖抑制实验
[0460]
实验目的:验证双-(10-去氧双氢青蒿素)-间苯三酚衍生物样品1、14~67对植物凝集素(pha)刺激的人外周血单个核细胞(pbmc)体外增值抑制活性。
[0461]
实验方法:分离人外周血单个核细胞(peripheral blood mononuclear cell,pbmc),调整细胞浓度为2
×
106/ml,每孔100μl种入96孔板中。将待测化合物溶解至合适浓度的母液,以1:3梯度稀释,得到5个浓度梯度(2、0.5、0.125、0.031和0.008μm),加入到对应的孔中,每个浓度设置三复孔,阳性对照孔为2μm环孢霉素a(csa),阴性对照孔为与实验组dmso含量一致的rpmi-1640完全培养基,同时每孔加入1μg/ml pha刺激pbmc细胞。培养箱中孵育72小时后,每孔加入ctg,检测细胞的增殖情况。分析软件graphpad prism 6.0处理数据。
[0462]
实验结果:ic
50
结果见表2
[0463]
表2化合物1及14~67对pha刺激人pbmc体外增殖抑制活性实验ic
50
(μm)
[0464][0465][0466]
从表2的数据可以发现,和化合物1相比,化合物14~67对pha刺激人pbmc体外增殖抑制活性实验ic
50
的数值与化合物1相当,说明化合物14~67保持了化合物1的免疫抑制活
性。
[0467]
实施例57化合物1、26、34、42、59和67抑制dnfb(2,4-二硝基氟苯)诱导的迟发性超敏反应的耳肿胀的药效研究实验方法:雌性icr小鼠按体重随机分成20组,每组10只。分别为模型组、阳性药dex(地塞米松,2mpk,qd)组、化合物1,26,34,42,59,67低中高各三个剂量给药组(2mpk,4mpk,8mpk)。模型诱导的方法是:所有组别在实验的第1天和第2天腹部涂抹50μl的1%dnfb溶液,在第6天时右耳两表面分别涂抹10μl的0.5%dnfb进行攻击,测量攻击前后的右耳厚度,利用差值对化合物的药效进行评价。实验分组及给药方案如表3所示:
[0468]
表3 耳肿胀实验分组及给药方案
[0469][0470][0471]
实验步骤:
[0472]
1.配制dnfb溶液:将丙酮和橄榄油(体积比为4:1,即4ml丙酮内加入1ml橄榄油)配制均匀的混合液,然后将dnfb加入丙酮和橄榄油的混合液中,分别配成1%和0.5%的dnfb溶液。
[0473]
2.致敏:所有动物麻醉、腹部剃毛(day 0)后,第1天(day 1)和第2天(day 2)在腹部剃毛部位涂抹50μl新鲜配制的1%dnfb溶液。
[0474]
3.第6天(day 6),所有动物测量右耳厚度数据,然后涂抹20μl的0.5%dnfb溶液(10μl/
[0475]
面)进行攻击。
[0476]
4.第7天(day 7)给药后再经过4小时,同时满足第6天(day 6)右耳涂抹0.5%dnfb经过了24小时后,用螺旋测微仪分别测每只小鼠右耳厚度,将右耳厚度减去第6天(day 6)攻击前右耳厚度所得差值做为肿胀值。
[0477]
5.给药:化合物(包括阳性药(dex))每天现配现用,口服,一天一次,连续给药7天。
[0478]
6.体重:每天给药前称量。
[0479]
统计分析:将各组收集数据用平均数和标准差表示(mean
±
sem)。对各种变化用graphpad prism软件进行分析。p《0.05认为有显著性差异,p《0.01认为有极显著差异。
[0480]
实验结果:实验的第6天(day 6),所有动物麻醉后测量右耳的厚度,测量三次,取平均值。实验的第7天(day 7),所有动物麻醉后测量右耳的厚度,测量三次,取平均值。将攻击前后右耳的厚度差数据进行统计分析。试验结束时,化合物1中剂量组(4mpk)小鼠死亡一半,高剂量组(8mpk)则无小鼠存活。攻击前后右耳的厚度差如图1所示(g1和g2-g20的统计学差异均显示g2-g20的p《0.05),与模型组相比,dex治疗组(2mpk)显著降低了小鼠攻击前后右耳的厚度差(p《0.0001),而五个化合物26、34、42、59和67的低中高三个剂量的治疗组也都显著降低了小鼠攻击前后右耳的厚度差(p《0.05),且有一定的剂量依赖效应。结果提示,化合物1显示出较高的毒性,动物不耐受;化合物26、34、42、59和67则动物耐受性良好,能够显著抑制dnfb诱导的耳肿胀,且药效与剂量呈相关性。
[0481]
实施例58化合物26、34、42、59和67抑制绵羊红细胞引起的迟发性超敏反应的跖肿胀药效研究
[0482]
实验方法:雌性icr小鼠按体重随机分成17组,每组10只;分别为模型组,醋酸泼尼松pns组(5mpk,qd),化合物26、34、42、59和67低中高各三个剂量给药组(2mpk,6mpk,18mpk)。所有组别在实验的第1天(day 1)腹腔注射绵羊红细胞,第5天(day 5)左跖足垫皮下注射绵羊红细胞进行攻击,在第6天(day 6)测量攻击后的左、右后跖足垫的厚度,利用差值对化合物的药效进行评价。实验分组及给药方案如表4所示:
[0483]
表4 跖肿胀实验分组及给药方案
[0484]
组别动物数量(只)给药方式化合物剂量(mg/kg/day)g1:模型组(纯水)10po,qd-g2:阳性药(pns)10po,qd5g3:化合物2610po,qd2g4:化合物2610po,qd6g5:化合物2610po,qd18g6:化合物3410po,qd2g7:化合物3410po,qd6g8:化合物3410po,qd18g9:化合物4210po,qd2
g10:化合物4210po,qd6g11:化合物4210po,qd18g12:化合物5910po,qd2g13:化合物5910po,qd6g14:化合物5910po,qd18g15:化合物6710po,qd2g16:化合物6710po,qd6g17:化合物6710po,qd18
[0485]
实验步骤:
[0486]
1.在第一天(day 1),g1-g17组所有动物腹腔注射2%(v/v)的srbc,每只注射0.2ml
[0487]
(约1
×
108个srbc),进行动物免疫。
[0488]
2.免疫后随即进行给药。
[0489]
3.在第5天(day 5),测量g1-g17组所有动物左后跖厚度,测量三次,取平均值;然后对g1-g17组所有动物测量部位皮下注射20%(v/v)的srbc,每只20μl(约1
×
108个
[0490]
srbc)进行攻击。
[0491]
4.攻击后18-20小时给药,24小时测量(day 6),所有动物麻醉后测量注射左后跖部位厚度,各测量3次,取平均值。以攻击后左后跖足垫厚度和右后跖足垫厚度差作为足肿胀度,分别予以数据分析。
[0492]
5.给药:化合物每天现配现用;口服;一天一次,连续给药6天。
[0493]
6.体重:每天给药前称量。
[0494]
统计分析:将各组收集数据用平均数和标准差表示(mean
±
sem)。对各种变化用graphpad prism软件进行分析。p《0.05认为有显著性差异,p《0.01认为有极显著差异。
[0495]
实验结果:实验的第6天,所有动物麻醉后测量左右后跖足垫的厚度,测量3次,取平均值。将左后跖足垫厚度和右后跖足垫厚度差数据进行统计分析。左右足跖的厚度差如图2所示(g1和g2-g17的统计学差异均显示g2-g17的p《0.05),与模型组相比,化合物26、34、42、59和67的低中高三个剂量治疗组也都显著降低了小鼠左右足跖的厚度差(p《0.05),有显著统计学差异。结果提示,化合物26、34、42、59和67能够显著抑制绵羊红细胞引起的跖肿胀迟发超敏反应,且药效与剂量相关。
[0496]
以上所述实施例仅表达了本发明的实施方式,其描述较为具体和详细,但并不能因此而理解为对本发明专利范围的限制。应当指出的是,对于本领域的普通技术人员来说,在不脱离本发明构思的前提下,还可以做出若干变形和改进,这些都属于本发明的保护范围。因此,本发明专利的保护范围应以所附权利要求为准。
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