CN104030904B - 4‑芳甲基姜黄素类似物作为Hsp90抑制剂的应用 - Google Patents

4‑芳甲基姜黄素类似物作为Hsp90抑制剂的应用 Download PDF

Info

Publication number
CN104030904B
CN104030904B CN201410249264.0A CN201410249264A CN104030904B CN 104030904 B CN104030904 B CN 104030904B CN 201410249264 A CN201410249264 A CN 201410249264A CN 104030904 B CN104030904 B CN 104030904B
Authority
CN
China
Prior art keywords
compound
arylmethyl
curcumin
hsp90 inhibitor
hsp90
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201410249264.0A
Other languages
English (en)
Other versions
CN104030904A (zh
Inventor
刘洋
许建华
叶敏
吴丽贤
吴群丹
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Fujian Medical University
Original Assignee
Fujian Medical University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Fujian Medical University filed Critical Fujian Medical University
Priority to CN201410249264.0A priority Critical patent/CN104030904B/zh
Publication of CN104030904A publication Critical patent/CN104030904A/zh
Application granted granted Critical
Publication of CN104030904B publication Critical patent/CN104030904B/zh
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C49/00—Ketones; Ketenes; Dimeric ketenes; Ketonic chelates
    • C07C49/20—Unsaturated compounds containing keto groups bound to acyclic carbon atoms
    • C07C49/255—Unsaturated compounds containing keto groups bound to acyclic carbon atoms containing ether groups, groups, groups, or groups
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D213/00—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members
    • C07D213/02—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members
    • C07D213/04—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D213/24—Heterocyclic compounds containing six-membered rings, not condensed with other rings, with one nitrogen atom as the only ring hetero atom and three or more double bonds between ring members or between ring members and non-ring members having three double bonds between ring members or between ring members and non-ring members having no bond between the ring nitrogen atom and a non-ring member or having only hydrogen or carbon atoms directly attached to the ring nitrogen atom with substituted hydrocarbon radicals attached to ring carbon atoms
    • C07D213/44—Radicals substituted by doubly-bound oxygen, sulfur, or nitrogen atoms, or by two such atoms singly-bound to the same carbon atom
    • C07D213/46—Oxygen atoms
    • C07D213/50—Ketonic radicals

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Abstract

本发明公开一种4‑芳甲基姜黄素类似物作为Hsp90抑制剂的应用，所述4‑芳甲基姜黄素类似物为如下结构式式中Ar为芳环或芳杂环，R1,R2,R3中至少有一个为非氢取代基。所述的4‑芳甲基姜黄素类似物在制备Hsp90抑制剂中的应用。所述的4‑芳甲基姜黄素类似物作为Hsp90抑制剂在制备治疗乳腺癌药物中的应用。

Description

4-芳甲基姜黄素类似物作为Hsp90抑制剂的应用
技术领域
本发明涉及一类化合物4-芳甲基姜黄素类似物，尤其是涉及一种作为Hsp90抑制剂在制备抗癌药物中的应用。
背景技术
热休克蛋白90（Hsp90）是ATP依赖的分子伴侣，能够稳定很多客户蛋白，其中有肿瘤生长增值所必需的许多蛋白，例如Her2，BCR-ABL 和IKK等。因此，Hsp90已成为重要的抗肿瘤靶点。Hsp90抑制剂可以抑制客户蛋白和辅伴侣的功能从而抑制多个癌症相关靶点（1、L. Neckers, P. Workman, Clin Cancer Res.2012,18,64-76）。第一代天然Hsp90抑制剂有格尔德霉素（GA）， 根赤壳菌素Radicicol（RD）及它们的衍生物。第二代合成Hsp90抑制剂主要以嘌呤和间苯二酚为骨架，有17个已进入临床研究（2、S. Soga, S. Akinaga, Y.Shiotsu, Current Pharmaceutical Design. 2013, 19, 366-376）。但由于有毒性大，生物利用度和水溶性差等缺点，至今还没有一个Hsp90抑制剂被批准上市。发现新化学骨架的Hsp90抑制剂具有重要意义。
姜黄素（curcumin，Cur）为天然植物姜黄中的主要抗肿瘤活性成分（3.T.Esatbeyoglu, P. Huebbe, I.M.I.Erns, D.Chin, A.E.Wagner, G.Rimbach, Angew Chem Int Ed Engl, 2012, 52, 5308-5332）。姜黄素能上调Hsp70（4、L. S. Angelo, D.S. Maxwell, J. Y. Wu, D. Sun, D.H. Hawke, I. E. McCutcheon, J. M. Slopis, Z.h. Peng, W. G. Bornmann, R. Kurzrock Bioorg.Med.Chem. 2013,21,932-939），下调Hsp90客户蛋白如BCL-ABL（5、L. X. WU, J. H. XU, X.W. HUANG, K.Z.ZHANG, C. X.WEN,Y. Z. CHEN, Acta Pharmacol Sin. 2006,27,694-699）, NF-κB,AKT，分解 Hsp90 辅伴侣p23（6、J.H. Lee, I.K. Chung, Cancer Lett.2009, 290, 76-86），很可能是Hsp90抑制剂。虽然姜黄素具有安全低毒广谱的优点，但由于其水溶性差、体内代谢快、活性偏低和靶点过多等缺点影响其成药性（7、P. Anand, A. B. Kunnumakkara, R.A.Newman, B. B.Aggarwal. Mol Pharm . 2007, 4, 807-818）。设计合成姜黄素衍生物，提高其选择性、活性和成药性，具有重要的理论和实际意义。但目前未见文献报道姜黄素衍生物或类似物为Hsp90抑制剂。
本申请人在中国专利200810071179.4中公开了4-(4-羟基-3-甲氧基苯甲基)姜黄素（化合物1）制备方法和抗肿瘤应用，但未见文献公开报道该化合物为Hsp90抑制剂。本专利所保护化合物2-9经SCIfinder检索均为新化合物，部分化合物体外抗肿瘤活性比化合物1强5-10倍。
发明内容
本发明的目的在于提供一种4-芳甲基姜黄素类似物及其制备Hsp90抑制剂及抗癌药物中的应用。
本发明的目的是这样实现的，所述的式1结构的4-芳甲基姜黄素类似物
式1
式中Ar为芳环或芳杂环，R1,R2,R3中至少有一个为非氢取代基。
式（1）中的Ar芳环为羟基取代或烷氧基取代芳环；优选4-羟基-3-甲氧基苯基或3，4，5-三甲氧基苯基；Ar芳杂环为吡啶芳杂环，优选3-吡啶基。
式（1）中的 R1,R2,R3为烷基,烷氧基，羟基，卤素或氢；优选异丙基，甲氧基，4-羟基，2-氟，2-氯。
所述的4-芳甲基姜黄素类似物为下表的化合物1-9；
本发明所述的4-芳甲基姜黄素类似物在制备Hsp90抑制剂中的应用。
本发明所述的4-芳甲基姜黄素类似物作为Hsp90抑制剂在制备治疗乳腺癌药物中的应用。
具体地说，本发明所述4-芳甲基姜黄素类似物的化学结构通式为：
其中Ar为芳环或芳杂环，R1,R2,R3中至少有一个为非氢取代基。式（Ⅰ）中的Ar所代表的基团为：取代苯基或杂环, 取代苯基优选羟基取代或烷氧基取代或卤代苯基最优选4-羟基-3-甲氧基苯基（见实施例4,5,6）和3，4，5-三甲氧基苯基（见实施例7,8,9）。所述的杂环优选吡啶基,吡咯基或呋喃基,最优选3-吡啶基（见实施例10,11,12）。4位苯甲基上的取代基可以是R1,R2,R3中的至少一个非氢取代基，或者有两个取代基如R2, R3，最多可以有三个取代基如R1,R2,R3，R1,R2,R3可以是相同的基团也可以是不同的基团，如可以R1= R2，也可以，R1≠ R2，所述的取代基优选烷基,烷氧基，卤素，卤代烷基或硝基,最优选甲氧基,羟基，异丙基,氟，三氟甲基或硝基；最优选的苯基上的取代基为2-氟（见实施例6,9,12）；2-氯（见实施例5,8,11）；4-羟基-3-甲氧基（见实施例4,7,10）。上述最优选的4-芳甲基姜黄素类似物即化合物1-9（见表1）。
本发明目的之二在于提供4-芳甲基姜黄素类似物作为Hsp90抑制剂在制备治疗乳腺癌药物的应用。
本发明的4-芳甲基姜黄素类似物，可用于但不局限于作为乳腺癌的药物。
根据本发明的实施方案，本发明所述的治疗癌症的4-芳甲基姜黄素类似物，可以治疗的癌症的例子包括但不局限于乳腺癌。
本发明的有益效果为：本发明的4-芳甲基姜黄素类似物体外均能显著抑制人乳腺癌肿瘤细胞的增殖,其中化合物4，7对SKBr3细胞抑制活性比化合物1强5倍和17倍，对MCF7细胞抑制活性比化合物1强4倍和3倍。从表2得到初步构效关系：Ar活性顺序从大到小为：3-吡啶基>3，4，5-三甲氧基>3-甲氧基-4-羟基；例如：固定4位取代苯甲基不变，对 SkBr3 细胞抑制活性化合物7(0.509μM)> 化合物4(1.66μM)> 化合物1 (8.55μM)；4位取代苯甲基上取代基活性顺序从大到小为：3-甲氧基-4-羟基>Cl>F。例如：固定Ar不变，对 SkBr3 细胞抑制活性化合物1 (8.55μM)> 化合物2(9.26μM)> 化合物3(10.2μM)；化合物4，7活性明显比化合物1强。本发明的4-芳甲基姜黄素类似物体外能够显著抑制人乳腺癌肿瘤细胞的增殖,见表2；说明本发明的4-芳甲基姜黄素类似物可用于作为制备治疗乳腺癌的药物。
附图说明
图1为化合物1对人乳腺癌细胞SKBr3中的Hsp70及其客户蛋白Her2表达的影响图。在图1中，显示化合物1作用SKBr3 24h后相关蛋白表达的western blot 分析结果。图1表明随着化合物1浓度（10, 20, 40μM）的增加，Hsp90的客户蛋白Her2在24h的作用时间下，呈现明显降解趋势，而Hsp70的表达量上调，即化合物1对Hsp90的客户蛋白有降解抑制作用。这一方式与阳性对照的Hsp90抑制剂NPU-AUY922（NPU,0.5μM）一致。DMSO(D)为阴性对照。
图2为化合物2，3，4，7，姜黄素，化合物1对人乳腺癌细胞SKBr3中的Hsp70及其客户蛋白Her2表达的影响图。在图2中，显示化合物1作用SKBr3 24h后相关蛋白表达的westernblot 分析结果。图2表明化合物2，3，4，7在5μM 浓度下，Hsp90的客户蛋白Her2在24h的作用时间下，呈现明显降解趋势，而Hsp70的表达量明显上调，即该类4-芳甲基姜黄素类似物对Hsp90的客户蛋白有降解抑制作用，而且明显比同一浓度的化合物1和姜黄素强。这一方式与阳性对照的Hsp90抑制剂NPU-AUY922（NPU,0.5μM）一致。DMSO(D)为阴性对照。
具体实施方式
下面结合实施例对本发明进行详细说明：
熔点用上海精密科学仪器厂WRS-1B型数字熔点仪测定, 温度未经校正。1H、13CNMR谱采用Bruker Avance Ⅲ型核磁共振仪(400 MHz)测定, TMS为内标。质谱采用Agilent6410 Triple Quad LC/MS液质联用仪测定。所用原料均购自国药化学试剂公司。
1)芳甲基姜黄素的合成通法
本发明的化合物可由乙酰丙酮先与香草醛Knovenagel缩合、再催化氢化得到中间体a，b，c后，中间体与二倍量的芳醛Claisen-Schimidt缩合得到4-芳甲基姜黄素类似物。合成路线和反应条件如下式：
()acetylacetone,piperdine,ethanol,室温,48h;()H2, Pd/C,acetone, 室温1h;()1.B2O3,EtOAc,40°C,0.5h;
2.ArCHO,B(n-BuO)3,n-BuNH2,40°C,4h,rt,overnight;3.HCl ,60°C,1h。
表1 实施例一～十二所合成的中间体a，b，c和化合物1～9的化学结构式
化合物 R1 R2 R3 Ar
a H OCH3 OH
b Cl H H
c F H H
1 H OCH3 OH 4-hydroxy-3-methoxyphenyl
2 Cl H H 4-hydroxy-3-methoxyphenyl
3 F H H 4-hydroxy-3-methoxyphenyl
4 H OCH3 OH 3,4,5-trimethoxyphenyl
5 Cl H H 3,4,5-trimethoxyphenyl
6 F H H 3,4,5-trimethoxyphenyl
7 H OCH3 OH 3-pyridine
8 Cl H H 3-pyridine
9 F H H 3-pyridine
表1中的中间体 a,b,c 合成通法
250ml单口烧瓶中加入乙酰丙酮10g(100mmol)，乙醇100ml，香草醛或2-氯苯甲醛或2-氟苯甲醛(100mmol)，滴加入催化量哌啶，室温下搅拌反应48小时，浓缩移去溶剂，乙醇重结晶。取第一步产品10g，溶于丙酮100ml，10％Pd/C 1g，Pd/C指钯/碳催化剂，在Parr1100催化氢化反应器中反应1h后，乙醚重结晶得中间体a，b，c。
实施例一 3-(4-羟基-3-甲氧苯甲基) 戊烷-2,4-二酮(中间体a)
白色固体, 熔点65-68°C。 1H NMR (CDCl3) δ: 2.08(s, 3H), 2.13(s, 3H),3.08(d, J =7.2Hz, 1H), 3.58(s, 1H), 3.86 (s, 3H), 4.00(t, 1H), 6.80-6.72 (m,2H), 7.07-6.98(m, 2H). 13C NMR (CDCl3) δ:23.26, 29.81, 32.02, 33.55, 55.89,70.18, 108.84, 115.60, 115.67, 128.49, 129.77, 131.35, 154.34, 154.70,192.23, 204.48; MS m/z: 237.1(M+H+).
实施例二 3-(2-氯苯甲基) 戊烷-2,4-二酮(中间体b)
淡灰色油状物。1H NMR (CDCl3) δ:2.03 (s, 3H), 2.16 (s, 3H), 3.24 (d, J =7.6 Hz, 1H), 3.70 (s, 1H), 4.15(t, 1H), 7.22-7.14 (m, 2H), 7.40-7.30 (m, 2H).13C NMR (CDCl3) δ: 23.03, 29.86, 30.54, 31.92, 66.98, 106.84, 127.06, 127.42,127.87, 128.36, 128.70, 129.70, 131.51, 134.06, 135.79, 136.93, 192.11,203.18. MS m/z :247.0 (M+Na+)
实施例三 3-(2-氟苯甲基) 戊烷-2,4-二酮(中间体c)
淡灰色油状物。1H NMR (CDCl3) δ:2.07 (s, 3H), 2.16 (s, 3H), 3.17 (d, J =7.6 Hz, 1H), 3.65 (s, 1H), 4.15(t, 1H), 7.15-7.00 (m, 2H), 7.20-7.15 (m, 2H).13C NMR (CDCl3) δ:23.12, 25.78, 27.74, 29.63, 67.94, 106.76, 115.10, 115.32,115.54, 124.29, 125.02, 126.56, 127.98, 128.58, 131.29, 159.79, 162.23,192.03, 203.25. MS m/z: 231.1(M+Na+)
2）化合物1-9合成通法
中间体a，b或c(10mmol)与三氧化二硼0.49g(7mmol)，乙酸乙酯20ml在100ml单口烧瓶中40℃搅拌反应0.5h。加入香草醛或3，4，5-苯甲醛或3-吡啶甲醛(20mmol)、硼酸三正丁酯11ml(40.5mmol)，室温下搅拌反应0.5小时，滴加入正丁胺0.2ml(2mmol)，0.5h内加完，室温下搅拌反应过夜。反应液加入0.5M盐酸15ml，升温至60℃反应1小时，用乙酸乙酯萃取，水洗，酯层干燥浓缩后过硅胶柱，得化合物1-9。
实施例四 4-(4-羟基-3-甲氧苯甲基)姜黄素(1)
橘黄色粉末, 熔点 203-205°C. 1HNMR(D6-DMSO): δ=3.71(s, 3H), 3.83(s,6H), 4.00(s, 2H), 6.61(d, 1H,J =8Hz), 6.69(d, 1H,J =8Hz), 6.80(d, 2H,J =8Hz),6.97(s, 1H), 7.19-7.14(m, 4H), 7.28(s, 2H), 7.59(d, 2H, J =15.2Hz), 8.74(s,1H), 9.65(s, 2H); 13C (D6-DMSO): δ=55.32, 55.59, 55.69, 111.37,111.43, 112.35,115.63, 118.12, 119.91, 122.64, 123.29, 123.70, 125.66, 126.63, 133.12,141.57, 144.13, 144.62, 147.23, 147.49, 147.92, 149.31, 149.79, 183.15,194.53. MS m/z: 503.5(M-1)。
实施例五 4-(2-氯苯甲基)姜黄素(2)
黄色粉末, 熔点 150.9-151.5°C. 1H NMR (CDCl3): δ=3.88 (s, 6H), 4.05 (s,2H), 6.69(d, J = 16 Hz, 1H), 6.74(d, J = 15.6 Hz, 1H), 6.91-6.87 (m, 2H),7.14-6.97 (m, 6H), 7.18 (s, 2H), 7.58 (d, J = 15.6 Hz, 1H), 7.70 (d, J = 15.2Hz, 1H). 13C NMR (CDCl3): δ=18.42, 28.61, 55.96, 56.04, 108.13, 109.58,109.77, 114.78, 114.84, 118.02, 121.84, 123.35, 124.14, 126.73, 126.93,127.53, 127.83, 127.91, 128.17, 129.23, 129.56, 129.96, 131.76, 133.01,136.30, 138.47, 142.30, 145.07, 146.78, 146.83, 147.98, 148.72, 183.59,194.42. MS m/z:491.1(M-1)。
实施例六 4-(2-氟苯甲基)姜黄素(3)
橙黄色粉末, 熔点 127.3-129.8°C. 1HNMR(CDCl3): δ=3.84(s, 3H), 3.86(s,3H), 3.90 (s, 2H), 6.62(d, 1H, J =16Hz), 6.78(d, 1H, J =15.2Hz), 7.03-6.82(m,6H), 7.16-7.07(m, 2H), 7.19 (s, 2H), 7.51(d, 1H, J =16Hz), 7.63(d, 1H, J =15.2Hz). 13C NMR(CDCl3): δ= 55.92, 56.04, 108.95, 109.70, 114.81, 118.07,123.12, 124.17, 127.97, 142.20, 145.08, 146.76, 147.93, 183.54. MS m/z:475.1(M-1)。
实施例七 (1E,6E)-4-(4-羟基-3-甲氧苯甲基)-1,7-双(3,4,5-三甲氧基)1,6-庚二烯-3,5-二酮(4)
橙红色粉末, 熔点 138.7-140.4°C. 1H NMR (CDCl3) :δ=3.82 (s, 3H), 3.87(s, 18H), 3.93 (s, 2H), 6.68 (s, 4H), 6.72(d, J = 15.6 Hz, 2H),6.90-6.86 (m,2H), 7.13 (s, 1H), 7.53(d, J = 7.6 Hz, 1H), 7.68 (d, J = 15.6 Hz, 2H), 9.87(s, 1H). 13C NMR ( CDCl3): δ=31.51, 55.98, 56.19, 56.25, 60.99, 105.46,105.95, 106.80, 109.48, 110.24, 111.71, 114.57, 120.47, 121.56, 123.27,129.54, 130.35, 130.87, 132.96, 140.19, 140.92, 141.84, 144.35, 145.00,146.49, 147.05, 153.44, 183.48, 191.09, 194.70. MS m/z:591.3 (M-1)。
实施例八 (1E,6E)-4-(2-氯苯甲基)-1,7-双(3,4,5-三甲氧基)1,6-庚二烯-3,5-二酮(5)
橙红色粉末, 熔点168.8.-169.0°C.1H NMR (CDCl3): δ= 3.79 (s, 18H), 4.00(s, 2H), 6.65 (s, 4H), 6.76 (d, J = 15.6 Hz, 2H), 7.14-7.12 (m, 3H), 7.35 (d,J = 7.6 Hz, 1H), 7.61 (d, J = 15.2 Hz, 2H). 13C NMR (CDCl3) :δ= 18.19, 28.48,56.15, 57.91, 60.93, 105.49, 105.58, 119.71, 127.66, 127.95, 128.02, 129.31,130.05, 130.79, 132.92, 138.53, 140.17, 142.41, 153.41, 183.50. MS m/z: 578.9(M-1)。
实施例九 (1E,6E)-4-(2-氟苯甲基)-1,7-双(3,4,5-三甲氧基)1,6-庚二烯-3,5-二酮(6)
橙红色粉末, 熔点160.0-160.4°C. 1H NMR (CDCl3): δ=3.88 (s, 18H), 3.99(s, 2H), 6.72 (s, 4H), 6.93 (d, J = 15.6Hz, 2H), 7.10-7.08 (m, 3H), 7.55 (d,J = 7.6Hz, 1H), 7.69 (d, J = 15.2 Hz, 2H). 13C NMR (CDCl3): δ= 23.67, 23.71,56.19, 56.25, 60.99, 105.46, 105.95, 108.64, 114.93, 115.15, 115.45, 119.73,123.22, 124.21, 124.77, 125.29, 127.84, 128.16,128.54, 129.53, 130.13,130.76, 131.56, 140.26, 142.22,140.92, 145.03, 153.47, 183.37, 194.31. MS m/z:562.9 (M-1)。
实施例十 (1E,6E)-4-(4-羟基-3-甲氧苯甲基)-1,7-双(3-吡啶基)1,6-庚二烯-3,5-二酮(7)
橙黄色粉末, 熔点169.1-170.1°C.1H NMR (D6-DMSO) :δ= 3.35 (s, 2H), 3.71(s, 3H), 6.70 - 6.57(m, 2H), 6.97 (s, 1H), 7.22 (d, J = 16.2 Hz, 1H), 7.56 -7.44 (m,3H), 7.71 (dd, J = 15.9, 7.8 Hz, 2H), 8.18 (m, 2H), 8.59 (m, 2H),8.75 (d, J = 7.4 Hz, 1H), 8.88 (dd, J = 11.7, 1.9 Hz, 2H). 13C NMR (D6-DMSO):δ = 29.88, 56.05, 112.48, 112.85, 116.18, 120.39, 123.75, 124.46, 127.83,130.52, 131.19, 132.99, 135.23, 138.46, 140.60, 145.23, 147.98, 150.44,150.65, 151.16, 151.69, 183.54, 195.43. MS m/z: 413.1(M-1)。
实施例十一 (1E,6E)-4-(2-氯苯甲基)-1,7-双(3-吡啶基)1,6-庚二烯-3,5-二酮(8)
黄色粉末, 熔点127.5-127.7°C. 1H NMR (CDCl3): δ= 4.06 (s, 2H), 6.96 (d,J = 15.2 Hz, 2H), 7.14-7.13 (m, 1H), 7.21-7.16 (m, 2H), 7.31-7.29 (m, 2H),7.48-7.46 (m, 1H), 7.79-7.75(m, 4H), 8.55(s, 1H), 8.56(s, 1H), 8.70(s, 1H),8.72(s, 1H). 13C NMR (CDCl3): δ= 28.73, 108.78, 122.69, 123.80, 125.86,127.47, 128.14, 129.34, 129.72, 130.87, 131.84, 133.33, 134.42, 137.44,138.84, 141.07, 149.64, 149.84, 150.26, 150.72, 151.55, 183.26, 193.88. MS m/z: 401.1(M-1)。
实施例十二 (1E,6E)-4-(2-氟苯甲基)-1,7-双(3-吡啶基)1,6-庚二烯-3,5-二酮(9)
黄色粉末, 熔点131.4-131.8°C. 1H NMR (CDCl3): δ= 3.99 (s, 2H), 7.15-7.03 (m, 3H), 7.23-7.20 (m, 1H), 7.33-7.29 (m, 2H), 7.83-7.74(m, 4H), 8.56(s,1H), 8.57(s, 1H), 8.71(s, 1H), 8.73(s, 1H). 13C NMR (CDCl3): δ= 23.96 ,108.62, 115.51, 122.26, 123.81, 124.87, 125.69, 126.90, 127.05, 128.39,128.77, 129.51, 129.89, 130.93, 134.41, 134.62, 138.77, 141.13, 149.79,150.26, 150.72, 151.55, 159.17, 161.61, 183.21, 193.89. MS m/z:385.1(M-1)。
3）MTT法检测4-芳甲基姜黄素类似物对细胞增殖的抑制作用实验
实施例十三 MTT法检测4-芳甲基姜黄素类似物对细胞增殖的抑制作用实验
将SKBr3或MCF7细胞（10000个/孔)接入96孔培养板培养过夜，实验组分别加入不同浓度的4-芳甲基姜黄素类似物见表2（DMSO为空白对照）和姜黄素，对照组不加药。另设空白组(只加培养基，无细胞)，每组设三个平行孔，370C培养48h，加入5mg/ml 的MTT溶液 20μl/孔，继续培养4h后，离心弃上清，加入DMSO 150μl，振荡10min，充分裂解后，用全自动酶标仪(美国BIO-RAD公司生产)检测570nm处的吸光度(OD570)值。根据吸光度计算细胞生长抑制率。
细胞生长抑制率=[OD对照－OD实验]/[ OD对照－OD空白] ×100%
以同一药物的不同浓度对肿瘤细胞生长抑制率作图，可得到剂量反应曲线，根据线性回归方程求出该药物的半数抑制浓度IC50，即细胞存活率减少50%时的药物浓度。MTT法结果表明（见表2，表2中的2-9分别指化合物2-9），本发明的4-芳甲基姜黄素类似物体外均能显著抑制人乳腺癌肿瘤细胞的增殖,其中化合物4，7对SKBr3细胞抑制活性比化合物1强5倍和17倍，对MCF7细胞抑制活性比化合物1强4倍和3倍。从表2得到初步构效关系：Ar活性顺序从大到小为：3-吡啶基>3，4，5-三甲氧基>3-甲氧基-4-羟基；例如：固定4位取代苯甲基不变，对 SkBr3 细胞抑制活性化合物7(0.509μM)> 化合物4(1.66μM)> 化合物1 (8.55μM)；4位取代苯甲基上取代基活性顺序从大到小为：3-甲氧基-4-羟基>Cl>F。例如：固定Ar不变，对 SkBr3 细胞抑制活性化合物1 (8.55μM)> 化合物2(9.26μM)> 化合物3(10.2μM)；化合物4，7活性明显比化合物1强。
表2 4-芳甲基姜黄素类似物对乳腺癌细胞SKBr3和MCF7的抑制作用
4）化合物对Hsp90客户蛋白的降解作用
实施例十四 化合物1 对Hsp90客户蛋白的降解作用
用化合物1(10, 20, 40μM)处理人乳腺癌SKBr3细胞24h, 0.5μM 的NPU-AUY922作为阳性对照,等量的DMSO作为阴性对照。吸去培养液，收集SKBR3细胞， PBS漂洗1次,加入NP-40 细胞裂解液，4℃裂解30min, 12000rpm 离心 15min.取上清，用Pierce BCAProtein Assay Kit (Thermo Scientific, USA)测定蛋白质浓度，调整蛋白浓度一致，进行SDS-PAGE电泳。 再转膜到硝酸纤维素（PVDF）膜上。一抗室温封闭2h，TBST洗涤3次，每次10min，二抗室温孵育1h，TBST洗涤后用SuperSignal WestPico (Thermo Scientific,USA)显影剂显影观察。
不同浓度的化合物1处理SKBr3细胞24h后，相关蛋白表达的western blot 分析结果。图1表明随着化合物1浓度（10, 20, 40μM）的增加，Hsp90的客户蛋白Her2在24h的作用时间下，呈现明显降解趋势，而Hsp70的表达量上调，即化合物1对Hsp90的客户蛋白有降解抑制作用。这一方式与阳性对照的Hsp90抑制剂NPU-AUY922（NPU,0.5μM）一致。DMSO(D)为阴性对照。
实施例十五 化合物2，3，4，7对Hsp90客户蛋白的降解作用
用5μM的化合物2，3，4，7和姜黄素处理人乳腺癌SKBr3细胞24h, 0.5μM 的NPU-AUY922作为阳性对照,等量的DMSO作为阴性对照。吸去培养液，收集SKBR3细胞， PBS漂洗1次,加入 NP-40 细胞裂解液，4℃裂解30min, 12000rpm 离心 15min.取上清，用PierceBCA Protein Assay Kit (Thermo Scientific, USA)测定蛋白质浓度，调整蛋白浓度一致，进行SDS-PAGE电泳。 再转膜到硝酸纤维素（PVDF）膜上。一抗室温封闭2h，TBST洗涤3次，每次10min，二抗室温孵育1h，TBST洗涤后用SuperSignal WestPico (Thermo Scientific,USA)显影剂显影观察。图2表明化合物2，3，4，7在5μM 浓度下，Hsp90的客户蛋白Her2在24h的作用时间下，呈现明显降解趋势，而Hsp70的表达量明显上调，即该类4-芳甲基姜黄素类似物对Hsp90的客户蛋白有降解抑制作用，而且明显比同一浓度的化合物1和姜黄素强。这一方式与阳性对照的Hsp90抑制剂NPU-AUY922（NPU,0.5μM）一致。DMSO(D)为阴性对照。

Claims (3)

1.一种4-芳甲基姜黄素类似物，其特征在于，为如下中的化合物1、化合物2或化合物3
化合物1
化合物2
化合物3。
2.权利要求1所述的4-芳甲基姜黄素类似物在制备Hsp90抑制剂中的应用。
3.权利要求1所述的4-芳甲基姜黄素类似物作为Hsp90抑制剂在制备治疗乳腺癌药物中的应用。
CN201410249264.0A 2014-03-07 2014-06-06 4‑芳甲基姜黄素类似物作为Hsp90抑制剂的应用 Active CN104030904B (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201410249264.0A CN104030904B (zh) 2014-03-07 2014-06-06 4‑芳甲基姜黄素类似物作为Hsp90抑制剂的应用

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
CN201410082729.8 2014-03-07
CN201410082729 2014-03-07
CN2014100827298 2014-03-07
CN201410249264.0A CN104030904B (zh) 2014-03-07 2014-06-06 4‑芳甲基姜黄素类似物作为Hsp90抑制剂的应用

Publications (2)

Publication Number Publication Date
CN104030904A CN104030904A (zh) 2014-09-10
CN104030904B true CN104030904B (zh) 2017-01-18

Family

ID=51461929

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201410249264.0A Active CN104030904B (zh) 2014-03-07 2014-06-06 4‑芳甲基姜黄素类似物作为Hsp90抑制剂的应用

Country Status (1)

Country Link
CN (1) CN104030904B (zh)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106083704B (zh) * 2016-06-06 2018-07-27 福建医科大学 3,5-(E)-二芳亚甲基-N-环丙基哌啶-4-酮类化合物作为Hsp90抑制剂的应用

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8841326B2 (en) * 2004-02-12 2014-09-23 Stc.Unm Therapeutic curcumin derivatives
CN101434525B (zh) * 2008-06-05 2012-07-04 福建医科大学 4-(4-羟基-3-甲氧基苯甲基)姜黄素及其用于制备抗肿瘤药物的应用
CN101700995B (zh) * 2008-12-30 2013-06-05 中山大学 具有抗肿瘤作用的多芳乙烯取代beta-二酮类化合物

Also Published As

Publication number Publication date
CN104030904A (zh) 2014-09-10

Similar Documents

Publication Publication Date Title
Salem et al. Synthesis and characterization of some new coumarins with in vitro antitumor and antioxidant activity and high protective effects against DNA damage
Bala et al. Mannich bases: an important pharmacophore in present scenario
CN104119280B (zh) 含氨基类脲与端炔结构单元的嘧啶衍生物、制备方法及应用
Mete et al. Synthesis and carbonic anhydrase inhibitory activities of new thienyl-substituted pyrazoline benzenesulfonamides
Wu et al. Design, synthesis and biological evaluation of novel benzimidazole-2-substituted phenyl or pyridine propyl ketene derivatives as antitumour agents
Sribalan et al. Multiple biological activities and molecular docking studies of newly synthesized 3-(pyridin-4-yl)-1H-pyrazole-5-carboxamide chalcone hybrids
Ramadan et al. Synthesis and anti H5N1 activities of some novel fused heterocycles bearing pyrazolyl moiety
Abdellatif et al. Synthesis, characterization and biological evaluation of novel 4′-fluoro-2′-hydroxy-chalcone derivatives as antioxidant, anti-inflammatory and analgesic agents
CN107021945B (zh) 一种含哌嗪酰胺类杨梅素类衍生物及其制备方法
Duangdee et al. Design synthesis and anti-proliferative activity of some new coumarin substituted hydrazide–hydrazone derivatives
Yejella et al. A study of anti-inflammatory and analgesic activity of new 2, 4, 6-trisubstituted pyrimidines
Narayana Reddy et al. Synthesis, in vitro antiproliferative activity, antioxidant activity and molecular modeling studies of new carbazole Mannich bases
CN101434600B (zh) 姜黄素哌啶酮结构类似物及其用于制备抗肿瘤药物的应用
O. Salas et al. Promising 2, 6, 9-trisubstituted purine derivatives for anticancer compounds: Synthesis, 3D-QSAR, and preliminary biological assays
Dinakaran et al. Synthesis and biological evaluation of novel pyrimidine-2 (1 H)-ones/thiones as potent anti-inflammatory and anticancer agents
Kim et al. Discovery of Pyrimidine-and Coumarin-Linked Hybrid Molecules as Inducers of JNK Phosphorylation through ROS Generation in Breast Cancer Cells
Ghany et al. Design, synthesis, anti-inflammatory evaluation, and molecular modelling of new coumarin-based analogs combined curcumin and other heterocycles as potential TNF-α production inhibitors via upregulating Nrf2/HO-1, downregulating AKT/mTOR signalling pathways and downregulating NF-κB in LPS induced macrophages
CN104030904B (zh) 4‑芳甲基姜黄素类似物作为Hsp90抑制剂的应用
Zapevalova et al. Synthesis, molecular docking, in vitro and in vivo studies of novel dimorpholinoquinazoline-based potential inhibitors of PI3K/Akt/mTOR Pathway
Raj et al. In vitro biological activities of new heterocyclic chalcone derivatives
Prasad et al. Design, Synthesis, Antioxidant, and Anti‐Breast Cancer Activities of Novel Diethyl (alkyl/aryl/heteroarylamino)(4‐(pyridin‐2‐yl) phenyl) methylphosphonates
CN106083704A (zh) 3,5‑(E)‑ 二芳亚甲基‑N‑环丙基哌啶‑4‑酮类化合物作为Hsp90抑制剂的应用
Konieczny et al. Structural Factors Affecting Cytotoxic Activity of (E)‐1‐(Benzo [d][1, 3] oxathiol‐6‐yl)‐3‐phenylprop‐2‐en‐1‐one Derivatives
Farghaly et al. Eco-friendly synthesis and 2D-QSAR study of novel pyrazolines as potential anticolon cancer agents
Gomha et al. Synthesis, characterization, and molecular docking of novel bis‐thiazolyl thienothiophene derivatives as promising cytotoxic antitumor drug

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
C14 Grant of patent or utility model
GR01 Patent grant