国产日韩欧美一区二区三区三州_亚洲少妇熟女av_久久久久亚洲av国产精品_波多野结衣网站一区二区_亚洲欧美色片在线91_国产亚洲精品精品国产优播av_日本一区二区三区波多野结衣 _久久国产av不卡

404 Not Found


nginx
404 Not Found

404 Not Found


nginx
404 Not Found

404 Not Found


nginx
404 Not Found

404 Not Found


nginx
404 Not Found

404 Not Found


nginx
404 Not Found

404 Not Found


nginx

藤黃酸衍生物的合成及其抗腫瘤活性

2019-12-04 03:06:06易春蝶張丹妮尹俊林曾廣智樊保敏
關鍵詞:藤黃甲酯衍生物

姜 毅,易春蝶,張丹妮,尹俊林,曾廣智,樊保敏

(1.云南民族大學 民族藥資源化學國家民委-教育部重點實驗室,云南 昆明 650500;2 云南民族大學 化學與環(huán)境學院,云南 昆明 650500)

自進入21世紀以來,癌癥日趨成為人類健康的最大威脅之一,也成為眾多醫(yī)藥學家研究的焦點[1].在早期,以順鉑,環(huán)磷酰胺等為代表的細胞毒性藥物曾經(jīng)是癌癥患者的首選藥物.之后以紫杉醇[2-3]為代表的天然產(chǎn)物藥物也成為了眾多患者的福音.藤黃酸作為從我國傳統(tǒng)中藥藤黃中分離出的有效成分,以其良好的抗癌活性與廣闊的發(fā)展前景受到眾多研究人員的關注.

藤黃酸(gambogic acid,GA)是一種具有獨特橋環(huán)呫噸酮骨架的天然化合物(見圖1),提取自藤黃科植物藤黃樹分泌的樹脂(中藥藤黃)[4].研究表明其具有優(yōu)秀的抗癌活性[5-6],可以通過誘導腫瘤細胞凋亡[7]、抑制腫瘤細胞增殖[8]、誘導分化[9]和抗腫瘤轉(zhuǎn)移[10]等作用達到抗癌的效果.藤黃酸在發(fā)揮抗癌活性的同時,對于造血系統(tǒng)和白細胞的影響較小.為了克服藤黃酸水溶性差,血管刺激強,穩(wěn)定性差等缺點,研究人員對其進行了多種結構修飾.Zhang等[11]的研究表明9-位,10-位的雙鍵是藤黃酸的關鍵活性基團,對于誘導腫瘤細胞凋亡的活性具有重要作用;對30-羧基與6-酚羥基的酯化和酰胺化等改造基本不影響藤黃酸的腫瘤細胞毒活性,在30-位引入某些基團反而會提高活性.Wang等[12]對GA 衍生物的研究中發(fā)現(xiàn),針對C-35,C-39 位甲基修飾,C-32,C-33 和C-37,C-38 雙鍵改造,可以提高GA 的抗腫瘤活性.

氨基酸是構成蛋白質(zhì)的基本單位,而蛋白質(zhì)是人體細胞的重要組成部分,具有構建細胞,調(diào)節(jié)代謝,提供能量等作用.動物體內(nèi)有20種必需氨基酸,根據(jù)其側鏈性質(zhì)的不同,可分為四大類:非極性氨基酸、極性氨基酸、酸性氨基酸和堿性氨基酸.在藥物分子中引入不同種類的氨基酸,可以改變其水溶性和代謝速率等多種特性,同時多個氨基酸相連而成的多肽還具有特異性識別等作用[13].為了得到更具發(fā)展前景的藤黃酸藥物,我們對藤黃酸30位進行酰胺化,通過向其引入不同種類的氨基酸,合成得到了多個藤黃酸衍生物,并對其進行了體外抗腫瘤活性篩選,希望得到具有更好發(fā)展前景的分子.

1 研究方法

以藤黃酸為原料,將氨基酸甲酯通過酰胺化法與藤黃酸30位羧酸相連接.其中疏水性氨基酸大多數(shù)直接相連接,親水性氨基酸先通過二氯樹脂保護側鏈再與藤黃酸相連,隨后脫掉保護基以暴露活性側鏈.合成路線見圖2.試劑和反應條件分別為:(a)DMAP,EDCI,CH2Cl2,DIEA,r.t.,35%~57% (b)HBTU,Amino acid-Resin,DIEA,DMF,r.t.;(c)CH2Cl2,CF3COOH,r.t.,28%~49%.

2 實驗部分

2.1 儀器與試劑

BrukerAvance 400、BrukerAvanceIII HD 600型核磁共振儀(CDCl3或CD4O為溶劑);Agilent 6420型質(zhì)譜儀(ESI-MS);Molecular Devices SpectraMax i3x型多功能酶標儀;Thermo Scientific 3425型二氧化碳培養(yǎng)箱.藤黃酸原料購自瑞芬思生物科技有限公司;氨基酸原料購自薩恩化學技術(上海)有限公司;保護氨基酸購自淘普生物科技有限公司.DMEM培養(yǎng)基、胎牛血清、谷氨酰胺購自Biological Industries公司;三(羥甲基)氨基甲烷購自生工生物工程(上海)股份有限公司;磺酰羅丹明B購自上海阿拉丁生化科技股份有限公司;三氯乙酸購自薩恩化學技術(上海)有限公司;冰乙酸購自天津富寧精細化工有限公司.實驗所用試劑均為分析純.

2.2 30-甘氨酸甲酯-藤黃酸衍生物的合成

在反應管中加入 10 mL 的 CH2Cl2,然后加入 50 mg 藤黃酸以及 10 mg 甘氨酸甲酯,38.11 mg(2.5 eq)EDCl,4.8 mg(0.5 eq)DMAP,39.4 mmL (3 eq)DIEA.常溫下攪拌 4 h.用 1N鹽酸水溶液與 10 mL 蒸餾水分別洗滌3次.收集有機相,用無水硫酸鈉干燥并旋干,硅膠柱層析用V(二氯甲烷)∶V(甲醇)=90 ∶10.產(chǎn)物為黃色固體,產(chǎn)率43%.1H NMR (CDCl3,400 MHz)δ∶1.28 (s,3H,19-H),1.30~1.43 (m,3H,24-H),1.45 (s,3H,40-H),1.55 (s,3H,35-H),1.69 (s,3H,29-H),1.74 (s,3H,25-H),1.80 (s,3H,34-H),1.98~2.07 (m,2H,36-H),2.28~2.35 (dd,J=4.48 Hz,J=13.44 Hz,1H,21a-H),2.54 (d,J=9.76 Hz,1H,22-H),2.66~2.75 (m,1H,32-H),2.93(d,J=32 Hz,2H,26-H),3.16~3.24 (m,1H,31b-H),3.27~3.36 (m,1H,31a-H),3.46 ~ 3.51 (m,1H,11-H),3.73 (m,3H,Gly-OMe),3.97 (s,1H,Gly-OMe),3.98 (s,1H,Gly-OMe),5.0~5.08 (m,1H,37-H),6.62 (d,J=12 Hz,1H,4-H),6.77~6.83 (m,1H,27-H),7.56 (d,J=8 Hz,1H,10-H),8.0 (s,1H,NH),12.86 (s,1H,6-OH);13C NMR (CDCl3,100 MHZ)δ:17.6 (C-34),18.1 (C-40),18.4 (C-39),21 (C-31),22.6 (C-36),25.2 (C-21),25.6 (C-35),25.7 (C-39),27.9 (C-19),28.8 (C-24),28.9 (C-25),29.9 (C-26),40.8 (C-Gly),42.2 (C-20),46.9 (C-11),48.9 (C-22),5.23 (C-Gly),58.4 (C-23),83.6 (C-2),84 (C-13),90.9 (C-14),100.39 (C-7),102.7 (C-5),107.7 (C-17),115.7 (C-4),123.6 (C-32),124.8 (C-37),126.3 (C-3),121.9 (C-28),131.8 (C-38),131.9 (C-33),133 (C-9),134 (C-10),135 (C-27),157.4 (C-16),157.6 (C-6),161.6 (C-18),170.2 (C-30),178.9 (C-8),204 (C-12);ESI-MS (m/z):700.1[M+H]+.

2.3 30-丙氨酸甲酯-藤黃酸衍生物的合成

操作同2.2,產(chǎn)物為黃色固體,產(chǎn)率61%.1H NMR (CDCl3,400 MHz)δ:1.28 (s,3H,19-H),1.30~1.43(m,3H,24-H),1.30~1.43 (m,3H,Ala-H),1.45 (s,3H,40-H),1.56 (s,3H,35-H),1.64 (s,3H,39-H),1.66 (s,3H,24H),1.69 (s,3H,29-H),1.73 (s,3H,25-H),1.80 (s,3H,34-H),1.99 ~ 2.08 (m,2H,36-H),2.30 (dd,J=5.6 Hz,J=13.2 Hz,1H,21a-H),2.54(d,J=10 Hz,1H,22-H),2.75~2.85(m,1H,32-H),2.93 (d,J=31.6 Hz,2H,26-H),3.12~3.24 (m,1H,31b-H),3.25~3.36 (m,1H,31a-H),3.42~3.52 (m,1H,11-H),3.73 (m,3H,Ala-OMe),4.44~4.54 (m,1H,Ala-OMe),5.0~5.08 (m,1H,37-H),5.0~5.08 (m,1H,3-H),5.43~5.45 (m,1H,32-H),6.07 (t,1H,32-H),6.67 (d,J=12 Hz,1H,4-H),6.64~6.83 (m,1H,27-H),7.56 (d,J=8 Hz,1H,10-H),8.0 (s,1H,NH),12.89 (s,1H,6-OH);13C NMR (CDCl3,100 MHZ)δ:17.6 (C-34),17.7 (C-Ala),18.1 (C-40),21 (C-31),22.6 (C-36),25.6 (C-35),25.7 (C-39),25.2 (C-21),27.9 (C-19),28.8 (C-24),28.9 (C-25),29.9 (C-26),42.2 (C-20),46.9 (C-11),47.8 (C-Ala),48.9 (C-22),5.23 (C-Ala),53.4 (C-23),83.6 (C-2),84 (C-13),90.9 (C-14),100.39 (C-7),102.7 (C-5),107.7 (C-17),115.7 (C-4),123.6 (C-32),124(C-37),126.3 (C-3),121.9 (C-28),131.8 (C-38),131.9 (C-33),133 (C-9),134 (C-10),135 (C-27),157.4 (C-16),157.6 (C-6),161.6 (C-18),173.4 (C-30),178.9 (C-8),204 (C-12);ESI-MS (m/z):714.1[M+H]+.

2.4 30-纈氨酸甲酯-藤黃酸衍生物的合成

操作同2.2,產(chǎn)物為黃色固體,產(chǎn)率44%.1H NMR (CDCl3,400 MHz)δ:0.89~0.95 (M,6H,Val-H),1.25 (s,3H,19-H),1.28 (s,3H,24-H),1.44 (s,3H,40-H),1.56 (s,3H,35-H),1.64 (s,3H,39-H),1.66 (s,3H,24H),1.70 (s,3H,29-H),1.74 (s,3H,25-H),1.82 (s,3H,34-H),1.99~2.07 (m,2H,36-H),2.31(dd,J=6 Hz,J=12 Hz,1H,21a-H),2.52~2.55(m,1H,22-H),2.57~2.66 (m,1H,32-H),3.18~3.29 (m,1H,31b-H),3.43~3.51 (m,1H,11-H),3.70(m,3H,Val-OMe),4.40~4.51(m,1H,Ala-OMe),5.02~5.13 (m,1H,37-H),5.02~5.13 (m,1H,3-H),5.43~5.45 (m,1H,32-H),5.45~5.47 (m,1H,4-H),6.67(d,J=12 Hz,1H,4-H),6.64~6.83 (m,1H,27-H),7.56 (d,J=8 Hz,1H,10-H),8.0 (s,1H,NH),12.89 (s,1H,6-OH);13C NMR (CDCl3,100 MHZ)δ:17.6 (C-34),17.7 (C-Ala),18.1 (C-40),21 (C-31),22.6(C-36),25.6(C-35),25.7(C-39),25.2 (C-21),27.9(C-19),28.8(C-24),28.9(C-25),29.9(C-26),42.2(C-20),46.9(C-11),47.8(C-Ala),48.9(C-22),52.3(C-Ala),53.4(C-23),83.6(C-2),84(C-13),90.9(C-14),100.39(C-7),102.7(C-5),107.7(C-17),115.7(C-4),123.6(C-32),124.8(C-37),126.3(C-3),121.9(C-28),131.8(C-38),131.9(C-33),133(C-9),134(C-10),135(C-27),157.4 (C-16),157.6 (C-6),161.6 (C-18),172 (C-30),178.9(C-8),205 (C-12);ESI-MS(m/z):742.1[M+H]+.

2.5 30-異亮氨酸甲酯-藤黃酸衍生物的合成

操作同2.2,產(chǎn)物為黃色固體,產(chǎn)率38%.1H NMR (CDCl3,400 MHz)δ:0.90 (t,3H,Ile-CH3),1.10~1.26(m,3H,Ile-CH3),1.27(s,3H,19-H),1.30~1.43(m,3H,24-H),1.45 (s,3H,40-H),1.55(s,3H,35-H),1.69 (s,3H,29-H),1.74(s,3H,25-H),1.80(s,3H,34-H),1.98~2.07(m,2H,36-H),2.28~2.34(dd,J=4.0 Hz,J=13.6 Hz,1H,21a-H),2.54(d,J=7.2 Hz,1H,22-H),2.56~2.64(m,1H,32-H),3.23~3.32(m,2H,31-H),3.45~3.48(m,1H,11-H),3.69(s,3H,Ile-OMe),4.50(dd,J=4.4 HzJ=7.6 Hz,1H,Ile-OMe),5.0~5.08(m,1H,3-H),6.67(d,J=10.4 Hz,1H,4-H),6.76(d,J=12Hz,1H,27-H),7.56(d,J=7.2 Hz,1H,10-H);13C NMR(CDCl3,100 MHZ)δ:11.5 (C-Ile),15.7 (C-Ile),17.6(C-34),18.2(C-40),21.2(C-31),22.7(C-36),25.6(C-35),25.7(C-39),25.2(C-21),27.9(C-19),28.8(C-24),28.9(C-25),29.9(C-26),37.4(C-Gly),42.2(C-20),46.9(C-11),48.9(C-22),52.3(C-Ile),56.4(C-23),83.2(C-2),83.8(C-13),90.9(C-14),100.53(C-7),102.7(C-5),107.7(C-17),115.7(C-4),122.1(C-28),123.6(C-32),124.8(C-37),125.4(C-3),131.7(C-38),131.9(C-33),132.7(C-9),134.2(C-10),136.2(C-27),157.4(C-16),157.6(C-6),161.6(C-18),172(C-30),178.9(C-8),205.5(C-12);ESI-MS m/z:756.[M+H]+

2.6 30-苯丙氨酸甲酯-藤黃酸衍生物的合成

操作同2.2,產(chǎn)物為黃色固體,產(chǎn)率29%.1H NMR (CDCl3,400 MHz)δ:1.32~1.36 (m,3H,24-H),1.38~1.44 (m,3H,40-H),1.55 (s,3H,35-H),1.69 (s,3H,29-H),1.66~1.72 (m,3H,25-H),1.74 (s,3H,34-H),2.01~2.04 (m,2H,36-H),2.26~2.40 (m,1H,21a-H),2.57~2.69 (m,1H,22-H),2.66~2.75 (m,1H,32-H),3.07~3.27 (m,2H,31-H),3.49 (s,3H,Phe-OMe),3.7~3.76 (m,2H,Phe-CH2),3.98 (s,1H,Phe-OMe),5.0~5.08 (m,1H,37-H),6.62~6.69 (m,1H,4-H),6.77~6.83 (m,1H,27-H),7.04~7.24 (m,5H,Phe),7.49~7.58 (m,1H,10-H),8.0 (s,1H,NH),12.7~12.8 (m,1H,6-OH);13C NMR (CDCl3,100 MHZ)δ:17.6 (C-34),18.1 (C-40),20.8 (C-31),22.7 (C-36),25.6 (C-35),25.7 (C-39),27.2 (C-19),28.4 (C-24),29.3 (C-25),29.8 (C-26),39.7 (C-Phe),41.7 (C-20),46.8 (C-11),49.0 (C-22),51.2 (C-Phe),52.5 (C-Phe),55.3 (C-23),82 (C-2),88.9 (C-14),108.6 (C-17),115.7 (C-4),123.8 (C-32),127.3 (C-Phe),128.9 (C-37),129.3 (C-Phe),132 (C-33),136 (C-27),155.8 (C-16),157.5 (C-6),161.3 (C-18),173.7 (C-30);ESI-MS (m/z):790.2[M+H]+.

2.7 30-絲氨酸甲酯-藤黃酸的合成

操作同2.2,產(chǎn)物為黃色固體,產(chǎn)率32%.1H NMR (CDCl3,600 MHz)δ:1.27(s,3H,19-H),1.34 (s,3H,24-H),1.45(s,3H,40-H),1.54(s,3H,35-H),1.58(s,3H,29-H),1.65(s,3H,25-H),1.90~1.98(m,2H,36-H),2.23~2.28(dd,J=5.4 Hz,J=12 Hz,1H,21a-H),2.51 (d,J=9.24 Hz,1H,22-H),2.73~2.81 (m,1H,32-H),3.10~3.16(m,1H,31b-H),3.23~3.29(m,1H,31a-H),3.59~3.66 (m,1H,Ser-H),3.74(s,3H,Ser-H),3.79(dd,1H,J=12 Hz,J=3 Hz,Ser-OMe),3.84~3.80 (m,1H,Ser-OMe),5.4(d,J=12 Hz,1H,4-H),6.87~5.92(m,1H,27-H),7.47(d,J=6.6 Hz,1H,10-H);13C NMR (CDCl3,150 MHZ)δ:17.4(C-34),17.9(C-40),20.7(C-39),22.4(C-31),22.6(C-36),24.7(C-35),25.3(C-21),25.5(C-39),27.1(C-19),28.8(C-24),29.2(C-25),29.5(C-26),42.1(C-20),46.7(C-11),49.1(C-22),52.2(C-Ser),59.9(C-23),83.4(C-2),83.8(C-13),91.1(C-14),100.2(C-7),102.6(C-5),107.7(C-17),115.7(C-4),123.5(C-32),124.7(C-37),126.43(C-3),131.6 (C-38),131.8(C-33),132.8(C-9),133.9(C-10),135.4(C-27),157.4(C-16),161.5(C-18),178.7(C-8);ESI-MS (m/z):730.1[M+H]+.

2.8 30-酪氨酸甲酯-藤黃酸的合成

操作同2.2,產(chǎn)物為黃色固體,產(chǎn)率36%.1H NMR(CDCl3,400 MHz)δ:1.29(s,3H,19-H),1.34(s,3H,24-H),1.37~1.44(s,3H,40-H),1.54(s,3H,35-H),1.62(s,3H,39-H),1.65(s,3H,24H),1.71(s,3H,29-H),1.73(s,3H,25-H),1.86(s,3H,34-H),1.99~2.06(m,2H,36-H),2.31(dd,J=4 Hz,J=14.8 Hz,1H,21a-H),2.54(d,J=9.6 Hz,1H,22-H),2.77~2.86(m,1H,32-H),2.87~2.95(m,2H,26-H),3.12~3.24(m,1H,31b-H),3.25~3.36(m,1H,31a-H),3.43~3.47(m,1H,11-H),3,49(s,2H,Tyr-OMe),3.73(m,3H,Tyr-OMe),4.98~5.12 (m,1H,37-H),4.98~5.12(m,1H,3-H),6.33(t,1H,32-H),6.61~6.66(m,1H,4-H),6.75~6.87(m,2H,Tyr-OMe),6.92~7.07(m,1H,27-H),7.11~7.22(m,2H,Tyr-OMe),7.51(d,J=7.6 Hz,1H,10-H),8.0(s,1H,NH),12.83(s,1H,6-OH);13C NMR (CDCl3,100 MHZ)δ:17.6 (C-34),18.1(C-40),21(C-31),22.6 (C-36),25.6(C-35),25.7(C-39),25.2(C-21),27.9(C-19),28.8(C-24),28.9(C-25),29.9(C-26),42.0(C-20),46.8(C-11),49.0(C-22),50.9(C-Tyr),52.1(C-Tyr),55.8(C-23),83.4(C-2),83.8(C-13),91.1(C-14),100.46(C-7),102.6 (C-5),107.7 (C-17),115.9(C-4),121.6(C-32),122.2(C-Tyr),124.5(C-37),130(C-Tyr),131.6(C-38),131.8(C-33),133.4(C-9),134.4(C-10),135.4(C-27),149.5(C-Tyr),157.5(C-16),157.6(C-6),161.6(C-18),175.2(C-30),178.9(C-8),203.6(C-12);ESI-MS(m/z):806.1[M+H]+.

2.9 30-色氨酸甲酯-藤黃酸的合成

操作同2.2,產(chǎn)物為黃色固體,產(chǎn)率55%.1H NMR (CDCl3,400 MHz)δ:1.26(s,3H,19-H),1.28~1.38(m,3H,24-H),1.42(s,3H,40-H),1.55(s,3H,35-H),1.62(s,3H,29-H),1.65(s,3H,25-H),1.72(s,3H,34-H),1.97~2.06(m,2H,36-H),2.17~2.29(m,1H,21a-H),2.39~2.50(m,1H,22-H),2.60~2.71(m,1H,32-H),3.14~3.24(m,1H,31b-H),3.24~3.33(m,1H,31a-H),3.14-3.33(m.2H,Trp-OMe),3.49(s,1H,11-H),3.64(s,3H,Trp-OMe),5.01~5.08(m,1H,37-H),6.66(d,J=10 Hz,1H,4-H),6.77~6.95(m,1H,27-H),7.0~1.5(m,5H,Trp-OMe),7.5~7.61(m,1H,10-H),8.0(s,1H,NH),12.86(s,1H,6-OH);13C NMR(CDCl3,100 MHZ)δ:17.6(C-34),18.2(C-40),18.3(C-39),21(C-31),22.8(C-36),25.8(C-35),25.2(C-21),27.9(C-19),28.8(C-24),28.9(C-25),29.9(C-26),42.2(C-20),46.9(C-11),48.9(C-22),5.23(C-Trp),58.4(C-23),83.6(C-2),84(C-13),91(C-14),100.7(C-7),102.9(C-5),107.8(C-17),111.3(C-Trp),116.0(C-4),118.8(C-Trp),119.5(C-Trp),121.9(C-28),122.3(C-Trp),122.9(C-Trp),123.9(C-32),124.8(C-37),126.6(C-3),127.6(C-Trp),131.8(C-38),131.9(C-33),132.7(C-9),133.7(C-10),136.2(C-27),157.4(C-16),157.6(C-6),161.6(C-18),172.2(C-30),204.5(C-12);ESI-MS(m/z):829.2[M+H]+.

2.10 30-賴氨酸-藤黃酸衍生物的合成

取 0.25 g 二氯樹脂,加入 2 mL CH2Cl2浸泡 0.5 h 進行活化,過濾并用 2 mL DMF 洗滌3次.加入 100 mg保護氨基酸Fmoc-Lys(Boc)-OH與 0.5 mL DIEA,加入 2 mL DMF 作為溶劑常溫反應 2 h,過濾收集固體,之后加入 0.5 mL 哌啶與 1.5 mL DMF 反應 0.5 h,再次過濾,洗滌并收集固體,加入 50 mg 藤黃酸,40 mg HBTU,0.5 mL DIEA,常溫反應 2 h,過濾后向固體中加入含25% 三氟乙酸的DCM,常溫反應 4 h,過濾收集濾液,濃縮后葡聚糖凝膠柱層析分離產(chǎn)物,得到黃色固體,產(chǎn)率49%.1H NMR(CDCl3,400 MHz)δ:1.28(s,3H,19-H),1.30~1.43(m,3H,24-H),1.44(s,3H,40-H),1.54(s,3H,35-H),1.56(s,2H,Lys-NH2),1.57(s,2H,Lys),1.60(s,3H,39-H),1.62~1.64 (m,3H,24H),1.67(s,3H,29-H),1.68(s,3H,25-H),1.73~1.82(m,2H,36-H),2.26~2.27(m,1H,21a-H),2.50~2.55(m,1H,22-H),2.50~2.55(m,2H,Lys),2.79~2.83(m,1H,32-H),2.89~2.95(m,2H,26-H),3.13~3.19(m,2H,31-H),3.35~3.38(m,1H,11-H),4.96~5.04(m,1H,37-H),4.96~5.04(m,1H,3-H),6.55(dd,J=4.2 Hz,J=10.8 1H,4-H),7.46~7.49(m,1H,10-H);13C NMR(CDCl3,150 MHZ)δ:17.0(C-34),19.7(C-40),21.4(C-31),21.6(C-36),22.4(Lys-C),24.4(C-39),24.6(C-21),27.4(C-19),28.7(C-24),28.9(C-25),29.1(Lys),32.7(Lys-C1),41.8(C-20),48.8(C-22),83.3(C-2),83.8(C-13),90.9(C-14),100.2(C-7),102.4(C-5),107.4(C-17),115.4(C-4),123.6(C-32),124.7(C-37),131.8(C-38),131.3(C-33),133(C-9),136.4 (C-27),157.2(C-16),157.7(C-6),161.5(C-18),171.9(C-30),179.6(C-8);ESI-MS(m/z):757.1[M+H]+.

2.11 30-組氨酸-藤黃酸的合成

操作同2.10,產(chǎn)物為黃色固體,產(chǎn)率28%.1H NMR(CDCl3,600 MHz)δ:1.33(s,3H,19-H),1.30~1.43(m,3H,24-H),1.44(s,3H,40-H),1.53(s,3H,35-H),1.57(s,3H,29-H),1.64(s,3H,25-H),1.69(s,3H,34-H),1.88~1.96(m,2H,36-H),2.22~2.26(dd,J=4.2 Hz,J=13.2 Hz,1H,21a-H),2.47~2.51(m,1H,22-H),2.72~2.78(m,1H,32-H),2.83~2.93 (m,2H,26-H),3.07~3.12(m,1H,His-C1),3.22~3.28(m,1H,31b-H),3.33~3.38 (m,1H,31a-H),3.42~3.47(m,1H,11-H),3.90~3.96(m,1H,His-C),4.93~4.99(m,1H,37-H),5.39(d,J=10.8 Hz,1H,32-H),5.89(t,1H,32-H),6.52(d,J=10 Hz,1H,4-H),7.42~7.47(m,1H,10-H),7.79(s,1H,NH),8.61~8.71(m,1H,His-C),12.86(s,1H,6-OH);13C NMR (CDCl3,150 MHZ)δ:17.6(C-34),18.1(C-40),20.9(C-31),22.8(C-36),25.2(C-21),25.7(C-39),27.8(C-19),28.9(C-25),29.9(C-26),42.2(C-20),46.9(C-11),48.9(C-22),83.6 (C-2),84(C-13),90.9(C-14),102.7(C-5),107.7(C-17),115.7(C-4),123.6(C-32),124.8(C-37),125.3(C-3),121.9(C-28),131.6(C-38),131.9(C-33),133.3(C-9),134.6(C-10),135.4(C-27),137.4(C-His),157.4(C-16),157.5(C-6),161.1(C-18),168.2(C-30),179.2(C-8),203.7(C-12);ESI-MS (m/z):766.1 [M+H]+.

2.12 活性檢測

用sulfurhodamine B(SRB)法檢測合成所得化合物對4種人源腫瘤細胞株(人肺癌細胞A549,人黑色素瘤細胞 A375,人肝癌細胞 HepG2,人胃癌細胞 BGC823)的體外抗腫瘤活性,并以藤黃酸為對照化合物.結果表明,所有修飾后的衍生物針對不同的細胞系表現(xiàn)出了不同的抗癌活性,并且在針對于特定細胞系時,抗癌活性均獲得了保持(表1).

表1 不同化合物對4種腫瘤細胞株的半數(shù)抑制濃度(IC50) (μmol·L-1)

1)細胞培養(yǎng) A549、A375、BGC823和HeGP2 細胞使用含10%胎牛血清的DMEM培養(yǎng)基,培養(yǎng)在37 ℃,含5%CO2的細胞培養(yǎng)箱內(nèi).

2)細胞細胞毒性(SRB)實驗 細胞鋪入96孔板,分別設定空白對照組、陰性對照組、陽性對照組和實驗組,每個樣品設置至少3個復孔.培養(yǎng) 24 h后,分別加入梯度稀釋的樣品,繼續(xù)培養(yǎng) 48 h,之后加入50% 三氯乙酸溶液固定 1 h.棄固定液、洗板、晾干之后加入含1%醋酸的0.4% SRB染色液.洗板、晾干之后加入10 mmol/L Tris溶液溶解染料,在 515 nm 波長下檢測其吸光度并計算樣品對細胞生長的抑制作用.

3 結果與討論

20種常用氨基酸中選擇了10種有代表性的氨基酸來進行反應,分別包括非極性氨基酸,水溶性性氨基酸和堿性氨基酸.在使用含硫氨基酸進行反應時發(fā)現(xiàn)其產(chǎn)物并不穩(wěn)定,所以沒有對含硫氨基酸的衍生物進行檢測.同時在合成含胍基的精氨酸-藤黃酸衍生物與酸性氨基酸-藤黃酸衍生物過程中,產(chǎn)生了大量的副產(chǎn)物.故我們沒有對這兩類氨基酸衍生物進行合成與研究,只選取了10種氨基酸進行反應.

在合成極性氨基酸衍生物11、12時借助了多肽合成思路,利用二氯樹脂作為氨基酸C端的保護基減少了副產(chǎn)物,同時得到具有暴露的極性基團的產(chǎn)物.由于中間產(chǎn)物全部連接在樹脂上,只需過濾即可去除雜質(zhì),也可以方便地使用茚三酮顯色反應來監(jiān)測反應進程.使用硅膠柱層析對非極性氨基酸衍生物進行純化,在純化11、12、13時,發(fā)現(xiàn)較為嚴重的吸附與拖尾,產(chǎn)品損失過大同時難以得到純品,所以改用葡聚糖凝膠柱,利用產(chǎn)物與雜質(zhì)分子質(zhì)量的不同進行分離,去除了吸附的效果,得到了純度較好的產(chǎn)物.

如表1所示,在合成的多種藤黃酸-氨基酸衍生物中,發(fā)現(xiàn)含有非極性側鏈的衍生物活性較好,如化合物2、3、4,它們顯示出比藤黃酸對照品更低的半數(shù)抑制濃度,表明合成的這幾種衍生物具有更強的體外抗癌活性.同時較小的側鏈對生物活性保持更有利,推測原因可能是由于非極性側鏈與細胞融合更好,更易透過細胞膜,同時也減緩其代謝速率,保證其作用時間.含有共軛基團與極性基團的衍生物活性降低,如7~12,其針對幾種癌細胞的體外細胞毒活性較之藤黃酸對照品,均有不同程度的下降.此外,修飾后的衍生物對于不同的細胞,其活性表現(xiàn)出了明顯的差異性,在針對A375與HepG2 2種細胞株的活性測試中,衍生物均體現(xiàn)了較好的生物活性,而針對BGC823細胞株的活性則有下降.而在針對A549細胞株的測試中,具有極性基團的衍生物普遍活性下降,其IC50大多提高10倍以上,這表明這一類衍生物針對A549細胞株的細胞毒活性幾乎完全喪失.藤黃酸衍生物在針對不同細胞系表現(xiàn)出不同作用強度的現(xiàn)象值得我們繼續(xù)深入研究.

猜你喜歡
藤黃甲酯衍生物
離子交換樹脂催化合成苯甲酸甲酯
云南化工(2020年11期)2021-01-14 00:50:52
藤黃中藤黃酸和新藤黃酸的提取分離及藥理毒理研究
烴的含氧衍生物知識鏈接
新型螺雙二氫茚二酚衍生物的合成
合成化學(2015年10期)2016-01-17 08:56:26
賴氨藤黃酸對乳腺癌MCF-7細胞增殖和凋亡的影響及其作用機制
K/γ-Al2O3催化丙酸甲酯合成甲基丙烯酸甲酯
化工進展(2015年3期)2015-11-11 09:07:41
卡前列甲酯栓聯(lián)合鈣劑預防及治療產(chǎn)后出血的效果觀察
超導可視聯(lián)合卡前列甲酯用于早早孕無痛人流術的效果觀察
紫外分光光度法測定藤黃藥材中總藤黃酸含量
Xanomeline新型衍生物SBG-PK-014促進APPsw的α-剪切
404 Not Found

404 Not Found


nginx
404 Not Found

404 Not Found


nginx
404 Not Found

404 Not Found


nginx
404 Not Found

404 Not Found


nginx
404 Not Found

404 Not Found


nginx
会泽县| 沂水县| 增城市| 尼勒克县| 洛隆县| 伊春市| 新龙县| 东港市| 甘德县| 庐江县| 黔西县| 南郑县| 安远县| 胶南市| 石林| 于田县| 南城县| 景宁| 宝坻区| 盐山县| 石林| 息烽县| 保康县| 英山县| 徐汇区| 巨鹿县| 大荔县| 中牟县| 扬州市| 克拉玛依市| 松桃| 青州市| 吴旗县| 太仆寺旗| 剑阁县| 吴江市| 即墨市| 麻江县| 澄江县| 增城市| 沛县|