CN103254133A - 二芳基吡唑苯胺类化合物的制备及其在结肠癌治疗中的应用 - Google Patents
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Abstract
二芳基吡唑苯胺类化合物,其特征是它有如下通式:
Description
技术领域
本发明涉及二芳基吡唑苯胺类化合物及其制备方法与抗肿瘤药物。
背景技术
癌症是一种严重威胁人类健康的疾病,临床治愈率很低。几乎所有的癌症都是由于细胞的异常增殖引起的,与调控细胞周期相关基因、蛋白的突变和失活密切相关。因此阻断异常的细胞增殖对于肿瘤的治疗是一种非常重要的手段。
Aurora激酶是有丝分裂中重要的调控激酶,与细胞的正常生长、增殖密切相关。Aurora主要有三个家族成员:Aurora-A、Aurora-B和Aurora-C,其中Aurora-A最为重要,Aurora-A蛋白的异常高表达,会导致多种肿瘤的发生,所以Aurora-A激酶已成为肿瘤治疗的重要靶点。
目前有很多小分子Aurora-A激酶抑制剂,其中吡唑环是重要的骨架结构,在最早合成并处于二期临床研究的Aurora-A激酶抑制剂药物VX-680就包含吡唑骨架,吡唑环上的氮原子可以和Aurora-A激酶ATP结合位点有氢键作用,从而抑制Aurora-A激酶活性。苯胺是一种具有抗肿瘤、抗炎和抗菌广谱药物活性的结构,苯胺中的苯环能和Aurora-A激酶ATP结合位点疏水作用和π-Cation作用,同时仲胺中的氮原子也增加化合物与Aurora-A的氢键作用。综上所述,本文中我们设计合成了一系列未经报道的二芳基吡唑苯胺类化合物,这些新化合物,可能体现出吡唑环和苯胺在Aurora-A激酶抑制和抗癌中的协同性。我们测试了这批化合物的生物活性,并发现其对人源宫颈癌HCT116细胞具有良好的抑制作用,且能抑制Aurora-A激酶活性,引起细胞凋亡。
发明内容
本发明的目的在于提供一类新型二芳基吡唑苯胺类化合物以及它们的制备方法与用途。
本发明的技术方案如下:
1.二芳基吡唑苯胺类化合物,其特征是它有如下通式:
R1为:4-氯、硝基、甲基、甲氧基。
R2为:4-溴、氯、甲氧基、氧乙基。
2.一种制备上述的二芳基吡唑苯胺类衍生物的方法,它由下列步骤组成:
步骤1.将取代基苯乙酮(20mmol)、盐酸苯肼(25mmol)、无水乙酸钠(40mmol)分别溶于乙醇,磁力搅拌,50-60℃反应4h(TLC点板检测反应)。反应完毕后将反应液旋干,乙酸乙酯和水萃取2-3次,旋干乙酸乙酯层,得到黄色固体。
步骤2.先将DMF(8ml)和POCl3(6ml)混合冰浴15min,然后将步骤1产物溶于3ml DMF滴加至上述混合液中,恢复常温继续搅拌30min,至70-80℃反应5h,冷却至常温后,倒入冰水中,用50%NaOH调PH至7-8,抽滤并水洗3遍,烘干后得固体。
步骤3.将步骤2所得产物溶于丙酮(30ml),然后将亚氯酸钠(30mmol)和氨基磺酸(35mmol)的混合水溶液(50ml)滴加至上述混合液中,常温下反应5h,反应完毕后将反应液旋干,用乙酸乙酯分别和饱和硫代硫酸钠、水各萃取2次,旋干乙酸乙酯层,得到白色固体。
步骤4.将步骤3得到的白色固体,取1mmol溶于氯化亚砜(1ml),磁力搅拌,70-80℃油浴反应3h,反应完毕后将氯化亚砜旋干,溶于二氯甲烷中(2ml)。将不同取代基的苯胺(1mmol)、DMAP(1.5mmol)和K2CO3(4mmol)加入上述混合液,磁力搅拌,40-50℃反应6h(TLC检测反应进行程度)。反应完毕后将反应液旋干,用乙酸乙酯和水、饱和NaCl和饱和K2CO3各萃取2次,旋干乙酸乙酯层,用甲醇重结晶,得到纯净的化合物1-9。
本发明的二芳基吡唑苯胺类具有抑制HCT116细胞增殖的作用。因此本发明的二芳基吡唑苯胺衍生物可做潜在的抗结肠癌药物。
具体实施方式
实施例一:N-(4-氧乙基苯基)-3-(4-甲基苯)-1-苯基-1H-吡唑-4-苯胺(化合物1)的制备
将对甲基苯乙酮(20mmol)、盐酸苯肼(25mmol)、无水乙酸钠(40mmol)分别溶于乙醇,磁力搅拌,50-60℃反应4h(TLC检测反应进行程度)。反应完毕后将反应液旋干,乙酸乙酯和水萃取2-3次,旋干有机层,得固体。将DMF和POCl3(12ml)混合冰浴15min,然后将上述固体溶于3ml DMF并滴加至上述混合液中,逐渐转入常温至70-80℃反应5h。冷却至常温后,倒入冰水中,用50%NaOH调pH至7-8,抽滤并水洗3遍,烘干后得固体。所得产物溶于丙酮(30ml),然后将亚氯酸钠(30mmol)和氨基磺酸(35mmol)的混合水溶液(50ml)滴加至上述混合液中,常温下反应5h,反应完毕后将反应液旋干,用乙酸乙酯分别和饱和硫代硫酸钠、水各萃取2次,旋干乙酸乙酯层,得到白色固体。取1mmol白色固体溶于氯化亚砜(1ml),磁力搅拌,70-80℃油浴反应3h,反应完毕后将氯化亚砜旋干,溶于二氯甲烷中(2ml)。将对氧乙基苯胺(1mmol)、DMAP(1.5mmol)和K2CO3(4mmol)加入上述混合液,磁力搅拌,40-50℃反应6h(TLC检测反应进行程度)。反应完毕后将反应液旋干,用乙酸乙酯和水、饱和NaCl和饱和K2CO3各萃取2次,旋干乙酸乙酯层,用甲醇重结晶,得到纯净的白色晶体状目标化合物。产率78%,mp:190-192℃;1H NMR(300MHz,DMSO-d6,δppm):8.56(s,2H);7.78(s,2H);7.60(s,2H);7.48(s,2H);7.32(s,3H);7.19(d,J=8.70Hz,3H);6.94(s,2H);6.77(s,1H);3.50(s,3H);1.83(m,3H);1.98(s,3H).MS(ESI):397.18(C25H24N3O2[M+H]+).Anal.Calcd for C25H23N3O2:C,75.54;H,5.83;N,10.57;O,8.05.Found:C,75.42;H,5.54;N,10.73;O,8.15
实施例二:N-(4-氧乙基苯基)-3-(4-氧甲基苯)-1-苯基-1H-吡唑-4-苯胺(化合物2)的制备
制备方法同实施例一。以对氧甲基苯乙酮代替例一中的对甲基苯乙酮,得到白色晶体状目标化合物。产率68%,mp:232-234℃;1H NMR(300MHz,DMSO-d6,δppm):8.54(s,1H);7.60(d,J=8.12Hz,2H);7.54(d,J=9.23Hz,4H);7.50(m,3H);7.35(m,3H);6.84(d,J=8.79Hz,2H);4.01(dd,J=14.07Hz,2H);3.88(s,3H);1.39(t,J=6.97Hz,3H).MS(ESI):413.17(C25H24N3O3[M+H]+).Anal.Calcd for C25H23N3O3:C,72.62;H,5.61;N,10.16;O,11.61.Found:C,75.42;H,5.56;N,10.43;O,11.46
实施例三:N-(4-氯苯基)-3-(4-氯苯)-1-苯基-1H-吡唑-4-苯胺(化合物3)的制备
制备方法同实施例一。以对氯苯乙酮代替例一中的对甲基苯乙酮,以对氯苯胺代替例一中的对乙氧基苯胺,得到白色晶体状目标化合物。产率76%,熔点:187-188℃;1H NMR(300MHz,DMSO-d6,δppm):8.56(s,1H);7.72(m,4H);7.52(m,4H);7.29(m,7H).MS(ESI):408.28(C22H16Cl2N3O[M+H]+).Anal.Calcdfor C22H15Cl2N3O;C,64.72;H,3.70;Cl,17.37;N,10.29;O,3.92.Found:C,64.54;H,3.66;Cl,17.23;N,10.15;O,3.84.
实施例四:N-(4-氧乙基苯基)-3-(4-氯苯)-1-苯基-1H-吡唑-4-苯胺(化合物4)的制备
制备方法同实施例一。以对氯苯乙酮代替例一中的对甲基苯乙酮,得到白色晶体状目标化合物。产率75%,mp:202-203℃;1H NMR(300MHz,DMSO-d6,δppm):8.54(s,1H);7.73(m,4H);7.48(m,4H);7.32(m,3H);6.86(t,J=2.42Hz,2H);3.99(dd,J=6.54Hz,2H);1.37(t,J=6.92Hz,3H).MS(ESI):417.12(C24H21ClN3O2[M+H]+).Anal.Calcd for C24H20ClN3O2:C,68.98;H,4.82;Cl,8.48;N,10.06;O,7.66.Found:C,68.60;H,4.71;Cl,8.35;N,10.01;O,7.69.
实施例五:N-(4-苯基)-3-(4-硝基苯)-1-苯基-1H-吡唑-4-苯胺(化合物5)的制备
制备方法同实施例一。以对硝基苯乙酮代替例一中的对甲基苯乙酮,以苯胺代替例一中的对乙氧基苯胺,得到棕色晶体状目标化合物。产率82%,mp:198-199℃;
1H NMR(300MHz,DMSO-d6,δppm):8.47(s,1H);8.33(d,J=8.45Hz,2H);8.08(d,J=8.49Hz,2H);7.78(d,J=8.51Hz,2H);7.41(m,3H);7,39(m,4H);6.88(d,J=6.90Hz,2H).MS(ESI):384.12(C22H17N4O3[M+H]+).Anal.Calcd for C22H16N4O3:C,68.74;H,4.20;N,14.58;O,12.49.Found:C,68.87;H,4.08;N,14.43;O,12.48.
实施例六:N-(4-氯苯)-3-(4-硝基苯)-1-苯基-1H-吡唑-4-苯胺(化合物6)的制备
制备方法同实施例一。以对硝基苯乙酮代替例一中的对甲基苯乙酮,以对氯苯胺代替例一中的对乙氧基苯胺,得到棕色晶体状目标化合物。产率83%,mp:209-210℃;1H NMR(300MHz,DMSO-d6,δppm):8.49(s,1H);8.34(d,J=5.46Hz,2H);8.06(d,J=5.13Hz,2H);7.78(d,J=4.87Hz,2H);7.53(t,J=4.81Hz,2H);7,40(m,4H);7.29(m,2H).MS (ESI):418.08(C22H16ClN4O3[M+H]+).Anal.Calcd forC22H15ClN4O3:C,63.09;H,3.61;Cl,8.46;N,13.38;O,11.46Found:C,63.12;H,3.52;Cl,8.32;N,13.45;O,11.58.
实施例七:N-(4-溴苯)-3-(4-硝基苯)-1-苯基-1H-吡唑-4-苯胺(化合物7)的制备。
制备方法同实施例一。以对硝基苯乙酮代替例一中的对甲基苯乙酮,以对溴苯胺代替例一中的对乙氧基苯胺,得到棕色晶体状目标化合物。产率87%,mp:190-192℃;1H NMR(300MHz,DMSO-d6,δppm):8.51(s,1H);8.34(d,J=8.72Hz,2H);8.06(d,J=8.41Hz,2H);7.77(d,J=8.47Hz,2H);7.45(m,3H);7,39(m,5H).MS(ESI):462.03(C22H16BrN4O3[M+H]+).Anal.Calcd for C22H15BrN4O3:C,57.04;H,3.26;Br,17.25;N,12.09;O,10.36.Found:C,57.12;H,3.20;Br,17.31;N,12.11;O,10.30.
实施例八:N-(4-氧甲基苯)-3-(4-硝基苯)-1-苯基-1H-吡唑-4-苯胺(化合物8)的制备。
制备方法同实施例一。以对硝基苯乙酮代替例一中的对甲基苯乙酮,以对氧甲基苯胺代替例一中的对乙氧基苯胺,得到棕色晶体状目标化合物。产率79%,mp:235-236℃;1H NMR(300MHz,DMSO-d6,δppm):8.48(s,1H);8.33(d,J=5.12Hz,2H);8.08(d,J=4.86Hz,2H);7.77(s,2H);7.53(s,2H);6.90(s,3H);7.36(m,3H);3.89(s,3H).MS(ESI):414.13(C23H19N4O4[M+H]+).Anal.Calcd forC23H18N4O4:C,66.66;H,4.38;N,13.52;O,15.44.Found:C,66.72;H,4.27;N,13.38;O,15.49
实施例九:N-(4-氧乙基苯)-3-(4-硝基苯)-1-苯基-1H-吡唑-4-苯胺(化合物9)的制备。
制备方法同实施例一。以对硝基苯乙酮代替例一中的对甲基苯乙酮,得到棕色晶体状目标化合物。产率80%,mp:211-112℃;1H NMR(300MHz,DMSO-d6,δppm):8.53(s,1H);7.76(d,J=4.90Hz,2H);7.68(m,4H);7.50(d,J=3.51Hz,3H);6.84(d,J=5.23Hz,1H);7.24(m,2H);4.02(d,J=4.23Hz,2H);1.40(t,J=4.30Hz,2H).MS(ESI):428.15(C24H21N4O4[M+H]+).Anal.Calcd for C24H20N4O4:C,67.28;H,4.71;N,13.08;O,14.94.Found:C,67.34;H,4.86;N,13.18;O,14.87
实施例十一:二芳基吡唑苯胺类化合物对肿瘤抑制活性研究
采用MTT[3-(4,5)-双甲基-2-噻唑-(2,5)-苯基溴化四氮唑蓝]法来测定二芳基吡唑苯胺类化合物对MCF7半抑制浓度,即IC50。
(1)培养液(每升)的配制:①悬浮细胞:RPMI-1640培养粉一袋(10.4g),新生牛血清100ml,青霉素溶液(20万U/ml)0.5ml,链霉素溶液(20万U/ml)0.5ml,加三蒸水溶解后,用5.6%的NaHCO3溶液调PH值至7.2-7.4,最后定容至1000ml。过滤灭菌。②贴壁细胞:同上,再加入NaHCO32.00g,HEPES 2.38g。
(2)D-Hanks缓冲液(每升)的配制:NaCl 8.00g,KCl 0.40g,Na2HPO4·12H2O 0.06g,KH2PO40.06g,NaHCO30.35g。高压灭菌。
(3)胰蛋白酶液的配制:利用D-Hanks缓冲液配成浓度为0.5%胰蛋白酶液。过滤除菌。
(4)实验药液的配制:将测试样品用少量的三蒸水溶解配成储备液,一般按实验最高浓度的10倍配制储备液。根据化合物溶解性不同,可用三蒸水直接溶解,或用少量DMSO助溶,再加三蒸水溶解。DMSO在培养液中的浓度不宜过大,加药后的每孔细胞悬液中DMSO的终浓度一般不超过0.05%-0.1%。储备液保存于-20℃冰箱中备用。
(5)MCF-7的培养:为贴壁生长细胞,常规培养于RPMI-1640培养液内(含10%小牛血清、100U/ml链霉素),置37℃、5%CO2培养箱中培养,每隔3-4天传代一次。传代时先弃去原培养液,再用D-Hanks缓冲液洗涤;然后用0.5%胰蛋白酶消化30秒左右,加入少量新鲜培养液终止消化;吹打,使贴壁细胞从培养瓶壁上脱落下来;移取适量至新鲜培养瓶中,再补充新鲜培养液至原体积(培养液体积约为培养瓶容量的1/10)。
(6)细胞孵育:取对数生长期的上述肿瘤细胞,调细胞悬液浓度为2×104个/ml。在96孔培养板中每孔加细胞悬液100μl,置37℃,5%CO2培养箱中培养24h。培养24h后,分别按设计加入药液。
(7)加药:将测试药液按照最终浓度的浓度梯度分别加入到各个孔中,每个浓度设6个平行孔。实验分为药物试验组(分别加入不同浓度的测试药)、对照组(只加培养液和细胞,不加测试药)和空白组(只加培养液,不加细胞和测试药)。将加药后的96孔板置于37℃,5%CO2培养箱中培养48h。
(8)存活细胞的测定:在培养了48h后的96孔板中,每孔加MTT 40μl(用40μl PBS配成2.5mg/ml的MTT)。在37℃放置4h后,移去上清液。每孔加100μl提取液(10%SDS-5%异丁醇-0.01M HCl)。37oC孵育过夜,最后,利用自动酶标仪在570nm波长处检测各孔的光密度(OD值)。
抑制率的计算:细胞生长的抑制率按照下列公式计算:
生长抑制率=(1-存活率)×100%=[1-(OD实验-OD空白)/(OD对照-OD空白)]×100%(OD实验表示测试药物组的平均光密度,OD对照表示对照组的平均光密度,OD空白表示对照组的平均光密度)。
半数抑制浓度(IC50)定义为当50%的肿瘤细胞存活时的药物浓度。根据测定的光密度(OD值),制作细胞生长抑制率的标准曲线,在标准曲线上求得其对应的药物浓度。
表1.Compound 1-10对MCF-7细胞增值的抑制作用
Claims (4)
2.一种制备上述的二芳基吡唑苯胺类衍生物的方法,它由下列步骤组成:
步骤1.将取代基苯乙酮(20mmol)、盐酸苯肼(25mmol)、无水乙酸钠(40mmol)分别溶于乙醇,磁力搅拌,50-60℃反应4h(TLC点板检测反应)。反应完毕后将反应液旋干,乙酸乙酯和水萃取2-3次,旋干乙酸乙酯层,得到黄色固体。
步骤2.先将DMF(8ml)和POCl3(6ml)混合冰浴15min,然后将步骤1产物溶于3ml DMF滴加至上述混合液中,恢复常温继续搅拌30min,至70-80℃反应5h,冷却至常温后,倒入冰水中,用50%NaOH调PH至7-8,抽滤并水洗3遍,烘干后得固体。
步骤3.将步骤2所得产物溶于丙酮(30ml),然后将亚氯酸钠(30mmol)和氨基磺酸(35mmol)的混合水溶液(50ml)滴加至上述混合液中,常温下反应5h,反应完毕后将反应液旋干,用乙酸乙酯分别和饱和硫代硫酸钠、水各萃取2次,旋干乙酸乙酯层,得到白色固体。
步骤4.将步骤3得到的白色固体,取1mmol溶于氯化亚砜(1ml),磁力搅拌,70-80℃油浴反应3h,反应完毕后将氯化亚砜旋干,溶于二氯甲烷中(2ml)。将不同取代基的苯胺(1mmol)、DMAP(1.5mmol)和K2CO3(4mmol)加入上述混合液,磁力搅拌,40-50℃反应6h(TLC检测反应进行程度)。反应完毕后将反应液旋干,用乙酸乙酯和水、饱和NaCl和饱和K2CO3各萃取2次,旋干乙酸乙酯层,用甲醇重结晶,得到纯净的化合物。
3.根据权利要求2所述的二芳基吡唑苯胺类衍生物的制备方法。
4.权利要求1所述的二芳基吡唑苯胺类在结肠癌治疗中的应用。
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CN108349953A (zh) * | 2015-11-18 | 2018-07-31 | 孟山都技术公司 | 杀虫组合物和方法 |
CN109970654A (zh) * | 2019-05-21 | 2019-07-05 | 郑州大学 | 一系列取代2-苯基吡唑衍生物及其制备方法和应用 |
US11548875B2 (en) | 2018-12-12 | 2023-01-10 | University Of Washington | Inhibitor compounds for male contraception |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004043951A1 (en) * | 2002-10-24 | 2004-05-27 | Carex S.A. | Pyrazole derivatives as modulators of peroxisome proliferator activated receptors |
CN102249945A (zh) * | 2010-05-21 | 2011-11-23 | 复旦大学 | 水杨酰苯胺类化合物及其制备方法和用途 |
-
2012
- 2012-02-20 CN CN2012100379165A patent/CN103254133A/zh active Pending
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2004043951A1 (en) * | 2002-10-24 | 2004-05-27 | Carex S.A. | Pyrazole derivatives as modulators of peroxisome proliferator activated receptors |
CN102249945A (zh) * | 2010-05-21 | 2011-11-23 | 复旦大学 | 水杨酰苯胺类化合物及其制备方法和用途 |
Non-Patent Citations (1)
Title |
---|
-: "RN 380457-31-8 REGISTRY", 《STN REGISTRY》 * |
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---|---|---|---|---|
CN108349953A (zh) * | 2015-11-18 | 2018-07-31 | 孟山都技术公司 | 杀虫组合物和方法 |
US11548875B2 (en) | 2018-12-12 | 2023-01-10 | University Of Washington | Inhibitor compounds for male contraception |
CN109970654A (zh) * | 2019-05-21 | 2019-07-05 | 郑州大学 | 一系列取代2-苯基吡唑衍生物及其制备方法和应用 |
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