CN102796164B - 京都酚修饰的脂肪胺/醇衍生物及其制备方法和应用 - Google Patents
京都酚修饰的脂肪胺/醇衍生物及其制备方法和应用 Download PDFInfo
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Abstract
本发明公开了京都酚(Tyr-Arg)修饰的脂肪胺/醇衍生物及其制备方法和作为镇痛剂和抗炎剂的应用,京都酚修饰的脂肪醇/胺衍生物的结构为通式I即Tyr-Arg-XCH2(CH2)nCH3,式中X为NH或O;n为6、8、10、12、14或16。本发明采用小鼠热甩尾法模型评价了通式化合物的镇痛活性,采用小鼠耳肿胀模型评价了通式I化合物的抗炎活性,实验结果表明通式I代表的12种化合物具有优秀的镇痛作用,且在相同剂量下具有较好的抗炎活性,可应用于镇痛药和抗炎药的生产中。
Description
技术领域
本发明涉及一种人工合成的多肽化合物,特别是涉及一种对二肽的羧基端用脂肪胺、脂肪醇进行保护修饰而得到的化合物及其制备方法以及在制备镇痛抗炎剂药物中的应用。
背景技术
疼痛作为人类主要生命指征之一,具有重要的生物学意义。世界疼痛大会将疼痛确认为继呼吸、脉搏、体温和血压之后的“人类第5大生命指征”,众多患者正在忍受着疼痛的折磨。目前,临床上使用的镇痛药物都存在各自的毒副作用,例如阿片类镇痛药的成瘾性和水杨类镇痛药对胃肠道的刺激性,寻找镇痛药物是新药研究的热点之一。在镇痛剂设计中发明人认识到京都酚(Tyr-Arg)是一种有潜力的镇痛二肽,以及口服活性差。按照这种认识,发明人对二肽的羧基端用脂肪胺、脂肪醇进行保护修饰,形成了本发明。
发明内容
本发明要解决的第一个技术问题是,提供通式I中的化合物(5a-l):Tyr-Arg-XCH2(CH2)n CH3,式中,X为NH或O;式中,n为6、8、10、12、14或16。
对所合成的通式I中的化合物(5a-1)进行编号:5a中X=NH,n=6;5b中X=NH,n=8;5c中X=NH,n=10;5d中X=NH,n=12;5e中X=NH,n=14;5f中X=NH,n=16;5g中X=O,n=6;5h中X=O,n=8;5i中X=O,n=10;5j中X=O,n=12;5k中X=O,n=14;5l中X=O,n=16。
以上编码号只是为了方便描述,并无任何对发明的限制意义。
本发明所要解决的第二个技术问题是提供通式化合物5a-l的制备方法,该方法包括:
(1)浓硫酸、浓硝酸条件下,Arg硝基化得NG-Arg(NO2);
(2)弱碱性水二氧六环条件下,NG-Arg(NO2)与(Boc)2O反应制得Nα-Boc-NG-Arg(NO2);
(3)在DCC和HOBt存在下,Nα-Boc-NG-Arg(NO2)在无水THF中与脂肪胺或脂肪醇缩合成Nα-Boc-NG-Arg(NO2)-XCH2(CH2)nCH3;
(4)在含氯化氢的乙酸乙酯中,Nα-Boc-NG-Arg(NO2)-XCH2(CH2)nCH3脱Boc保护,得NG-Arg(NO2)-XCH2(CH2)nCH3;
(5)在DCC和HOBt存在下,Boc-Tyr在无水THF中与NG-Arg(NO2)-XCH2-(CH2)nCH3缩合,制备得到Boc-Tyr-NG-Arg(NO2)-XCH2(CH2)nCH3;
(6)H2/Pd条件下Boc-Tyr-NG-Arg(NO2)-XCH2(CH2)nCH3脱-NO2保护,得Boc-Tyr-Arg-XCH2(CH2)nCH3;
(7)在含氯化氢的乙酸乙酯中,Boc-Tyr-Arg-XCH2(CH2)nCH3脱Boc保护,即得Tyr-Arg-XCH2(CH2)nCH3。
该制备方法可以用图1的路线概括。
本发明所要解决的第三个技术问题是提供了所述的化合物在制备镇痛和抗炎药物中的应用。
本发明所要解决的第四个技术问题是在小鼠热甩尾法模型上评价本发明的化合物的镇痛活性。
本发明的第五个目的是在小鼠耳肿胀模型上评价本发明的化合物的抗炎活性。
附图说明
图1为本发明通式5a-l化合物的合成路线示意图。
i)DCC,HOBt,CH3(CH2)nCH2NH2(n=6,8,10,12,14,16)或CH3(CH2)nCH2OH,(n=6,8,10,12,14,16);ii)EtoAc/HCl,冰浴;iii)DCC,HOBt,Boc-Tyr;iv)H2,Pd/C;v)EtOAc/HCl,冰浴;5a中X=NH,n=6;5b中X=NH,n=8;5c中X=NH,n=10;5d中X=NH,n=12;5e中X=NH,n=14;5f中X=NH,n=16;5g中X=O,n=6;5h中X=O,n=8;5i中X=O,n=10;5j中X=O,n=12;5k中X=O,n=14;5l中X=O,n=16。
具体实施方式
为了进一步阐述本发明,下面给出一系列实施例。这些实施例完全是例证性的,它们仅用来对本发明进行具体描述,不应当理解为对本发明的限制。
实施例1制备NG-Arg(NO2)
冰盐浴搅拌条件下,向500ml茄瓶中加入80ml浓硝酸,然后缓慢加入80ml浓硫酸。10min后,分批加入47.33g(272mmol)L-Arg,反应混合物冰盐浴下搅拌反应2h。向反应混合物中缓慢加入冰水96ml,然后缓慢加入浓氨水,调pH=8,再用冰醋酸调pH=6,有大量无色固体析出,放入4℃冰箱中继续析晶。减压过滤,弃去滤液,滤饼用80℃热水溶解,放入4℃冰箱中重结晶。室温下减压过滤,收集不溶物52.5g,为无色固体,产率88%.ESI-MS(m/e):220[M+H]+。
实施例2制备Nα-Boc-NG-Arg(NO2)
称取1.752g(8mmol)NG-Arg(NO2)于反应瓶中,加入3ml水溶解,冰浴条件下,缓缓滴加1N NaOH 8ml,将溶有1.92g(Boc)2O的二氧六环缓缓倒入其中,调反应液pH为9,反应48h,期间每隔2h用单通抽反应体系中CO2一次。TLC检测反应完全后,调反应液pH为7,旋除二氧六环,再调反应液pH为2,用乙酸乙酯萃取反应液三次,得到的乙酸乙酯层再用饱和NaCl萃洗三次,得到乙酸乙酯层用无水NaSO4干燥过夜,过滤除去NaSO4,旋干乙酸乙酯得到标题化合物2.4g,为无色固体,产率94%.ESI-MS(m/e):320[M+H]+。
实施例3制备NG-Arg(NO2)-OBzl
取6.57g(30mmol)NG-Arg(NO2)于反应瓶中,然后加入6.21g苯磺酸钠,50ml苯甲醇,20ml环己烷,80℃油浴下反应8h.向反应液中加入大量乙醚,有无色固体析出,常温减压过滤,得标题化合物14.1g,为无色固体,产率98%.ESI-MS(m/e):310[M+H]+.
实施例4制备Boc-Tyr
称取0.905g(5mmol)Boc-Tyr于反应瓶中,加入2ml水溶解,冰浴条件下,缓缓滴加1N NaOH 5ml,将溶有1.199g(Boc)2O的二氧六环缓缓倒入其中,调反应液pH为9,反应48h,期间每隔2h用单通抽反应体系中CO2一次。TLC检测反应完全后,调反应液pH为7,旋除二氧六环,再调反应液pH为2,用乙酸乙酯萃取反应液三次,得到的乙酸乙酯层再用饱和NaCl萃洗三次,得到乙酸乙酯层用无水NaSO4干燥过夜,过滤除去NaSO4,旋干乙酸乙酯得到标题化合物0.91g,为无色固体,产率65%.ESI-MS(m/e):282[M+H]+.
实施例5制备Nα-Boc-NG-Arg(NO2)-NHCH2(CH2)6CH3(1a)
将3.06g(10.8mmol)Nα-Boc-NG-Arg(NO2)溶于20ml无水四氢呋喃(THF),冰浴下往里加1.08g(8mmol)N-羟基苯并三唑(HOBt)和1.98g(9.6mmol)二环己基羰二亚胺(DCC)的无水THF溶液。反应混合物冰浴搅拌30分钟,得对应的活泼酯溶液,待用。
将1.032g(8mmol)CH3(CH2)7NH2溶于20ml无水四氢呋喃(THF),然后与上面待用的活泼酯溶液混溶,用N-甲基吗啉(NMM)调pH=8~9。得到的反应混合物室温反应24小时。TLC(展开剂CHCl3∶CH3OH=30∶1)显示直链CH3(CH2)7NH2消失。反应混合物减压浓缩至干,残留物用乙酸乙酯溶解,滤除不溶物。滤液依次用5%碳酸氢钠水溶液洗3次,饱和氯化钠水溶液洗3次;5%硫酸氢钾水溶液洗3次,饱和氯化钠水溶液洗3次和饱和碳酸氢钠水溶液洗3次,饱和氯化钠水溶液洗3次。分出的乙酸乙酯层用无水硫酸钠干燥、过滤、滤液32℃减压浓缩,重结晶纯化,得3.01g标题化合物,为无色固体,产率88%.ESI-MS(m/e):453.6[M+Na]+.
实施例6制备Nα-Boc-NG-Arg(NO2)-NHCH2(CH2)8CH3 (1b)
按照实施例5的方法,2.42g(7.58mmol)Nα-Boc-NG-Arg(NO2)与0.976g(6.22mmol)CH3(CH2)9NH2反应,得到标题化合物2.52g,为无色固体,产率88%.ESI-MS(m/e):481.7[M+Na]+.
实施例7制备Nα-Boc-NG-Arg(NO2)-NHCH2(CH2)10CH3 (1c)
按照实施例5的方法,3.06g(9.59mmol)Nα-Boc-NG-Arg(NO2)与1.48g(8mmol)CH3(CH2)11NH2反应,得到标题化合物3.6g,为无色固体,产率93%.ESI-MS(m/e):509.5[M+Na]+.
实施例8制备Nα-Boc-NG-Arg(NO2)-NHCH2(CH2)12CH3 (1d)
按照实施例5的方法,3.06g(9.59mmol)Nα-Boc-NG-Arg(NO2)与1.704g(8mmol)CH3(CH2)12NH2反应,得到标题化合物4.0g,为无色固体,产率97%.ESI-MS(m/e):537.7[M+Na]+.
实施例9制备Nα-Boc-NG-Arg(NO2)-NHCH2(CH2)14CH3 (1e)
按照实施例5的方法,3.06g(9.59mmol)Nα-Boc-NG-Arg(NO2)与1.928g(8mmol)CH3(CH2)15NH2反应,得到标题化合物4.11g,为无色固体,产率95%.ESI-MS(m/e):565.3[M+Na]+.
实施例10制备Nα-Boc-NG-Arg(NO2)-NHCH2(CH2)16CH3 (1f)
按照实施例5的方法,3.06g(9.59mmol)Nα-Boc-NG-Arg(NO2)与2.152g(8mmol)CH3(CH2)17NH2反应,得到标题化合物4.4g,为无色固体,产率96%.ESI-MS(m/e):593.5[M+Na]+.
实施例11制备Nα-Boc-NG-Arg(NO2)-OCH2(CH2)6CH3 (1g)
将4.785g(15mmol)Boc-Nα-Boc-NG-(NO2溶于30ml无水四氢呋喃(THF),冰浴下往里加0.525g(15mmol)N-羟基苯并三唑(HOBt)和3.708g(18mmol)二环己基羰二亚胺(DCC)的无水THF溶液。反应混合物冰浴搅拌30分钟,得对应的活泼酯溶液,待用。
将2.34g(19.5mmol)CH3(CH2)7OH溶于20ml无水四氢呋喃(THF),然后与上面待用的活泼酯溶液混溶。得到的反应混合物室温反应24小时。TLC(展开剂CHCl3∶CH3OH=30∶1)显示Nα-Boc-NG-Arg(NO2)消失。反应混合物减压浓缩至干,残留物用乙酸乙酯溶解,滤除不溶物。滤液依次用5%碳酸氢钠水溶液洗3次,饱和氯化钠水溶液洗3次;5%硫酸氢钾水溶液洗3次,饱和氯化钠水溶液洗3次和饱和碳酸氢钠水溶液洗3次,饱和氯化钠水溶液洗3次。分出的乙酸乙酯层用无水硫酸钠干燥、过滤、滤液32℃减压浓缩,柱层析纯化,得2.0g标题化合物,为无色固体,产率31%.ESI-MS(m/e):454.7[M+Na]+.
实施例12制备Nα-Boc-NG-Arg(NO2)-OCH2(CH2)8CH3 (1h)
按照实施例11的方法,4.785g(15mmol)Nα-Boc-NG-Arg(NO2)与3.081g(19.6mmol)CH3(CH2)9OH反应,得到标题化合物2.0g,为无色固体,产率29%.ESI-MS(m/e):482.6[M+Na]+.
实施例13制备Nα-Boc-NG-Arg(NO2)-OCH2(CH2)10CH3 (1i)
按照实施例11的方法,4.785g(15mmol)Nα-Boc-NG-Arg(NO2)与3.627g(19.6mmol)CH3(CH2)11OH反应,得到标题化合物2.0g,为无色固体,产率27%.ESI-MS(m/e):510.5[M+Na]+.
实施例14制备Nα-Boc-NG-Arg(NO2)-OCH2(CH2)12CH3 (1j)
按照实施例11的方法,4.785g(15mmol)Nα-Boc-NG-Arg(NO2)与4.173g(19.6mmol)CH3(CH2)12CH2OH反应,得到标题化合物1.5g,为无色固体,产率19%.ESI-MS(m/e):538.7[M+Na]+.
实施例15制备Nα-Boc-NG-Arg(NO2)-OCH2(CH2)14CH3 (1k)
按照实施例11的方法,4.785g(15mmol)Nα-Boc-NG-Arg(NO2)与4.719g(19.6mmol)CH3(CH2)14CH2OH反应,得到标题化合物2.1g,为无色固体,产率26%.ESI-MS(m/e):566.5[M+Na]+.
实施例16制备Nα-Boc-NG-Arg(NO2)-OCH2(CH2)16CH3 (1l)
按照实施例11的方法,4.785g(15mmol)Nα-Boc-NG-Arg(NO2)与5.265g(19.6mmol)CH3(CH2)17OH反应,得到标题化合物2.0g,为无色固体,产率23%.ESI-MS(m/e):594.4[M+Na]+.
实施例17制备HCl·NG-Arg(NO2)-NHCH2(CH2)6CH3 (2a)
3.01g(8.2mmol)Nα-Boc-NG-Arg(NO2)-NHCH2(CH2)6CH3溶于适量乙酸乙酯,加入约25ml 4N无水氯化氢-乙酸乙酯液,室温搅拌3小时,TLC(展开剂石油醚∶丙酮=5∶1)显示原料点消失。反应混合液在室温下减压浓缩,残留物再用乙酸乙酯溶解并室温下浓缩,如此反复4次;然后向浓缩物中加入乙醚,室温下减压浓缩,如此反复3次,直至除净游离的氯化氢,得标题化合物2.6g,为无色固体,产率87%.ESI-MS(m/e):331.8[M+H]+.
实施例18制备HCl·NG-Arg(NO2)-NHCH2(CH2)8CH3 (2b)
按照实施例17的方法,以2.52g(5.50mmol)Nα-Boc-NG-Arg(NO2)-NHCH2(CH2)8CH3为原料,得标题化合物1.969g,为无色固体,产率91%.ESI-MS(m/e):359.7[M+H]+.
实施例19制备HCl·NG-Arg(NO2)-NHCH2(CH2)10CH3(2c)
按照实施例17的方法,以3.6g(7.41mmol)Nα-Boc-NG-Arg(NO2)-NHCH2(CH2)10CH3为原料,得标题化合物2.98g,为无色固体,产率95%.ESI-MS(m/e):387.5[M+H]+.
实施例20制备HCl·NG-Arg(NO2)-NHCH2(CH2)12CH3 (2d)
按照实施例17的方法,以4.1g(7.98mmol)Nα-Boc-NG-Arg(NO2)-NHCH2(CH2)12CH3为原料,得标题化合物3.33g,为无色固体,产率93%.ESI-MS(m/e):415.5[M+H]+.
实施例21制备HCl·NG-Arg(NO2)-NHCH2(CH2)14CH3 (2e)
按照实施例17的方法,以4.28g(7.9mmol)Nα-Boc-NG-Arg(NO2)-NHCH2(CH2)14CH3为原料,得标题化合物3.43g,为无色固体,产率90%.ESI-MS(m/e):443.5[M+H]+.
实施例22制备HCl·NG-Arg(NO2)-NHCH2(CH2)16CH3 (2f)
按照实施例17的方法,以4.4g(7.72mmol)Nα-Boc-NG-Arg(NO2)-NHCH2(CH2)16CH3为原料,得标题化合物3.84g,为无色固体,产率98%.ESI-MS(m/e):471.7[M+H]+.
实施例23制备HCl·NG-Arg(NO2)-OCH2(CH2)6CH3 (2g)
按照实施例17的方法,以2.0g(4.64mmol)Nα-Boc-NG-Arg(NO2)-OCH2-(CH2)6CH3为原料,得标题化合物1.68g,为无色固体,产率98%.ESI-MS(m/e):332.5[M+H]+.
实施例24制备HCl·NG-Arg(NO2)-OCH2(CH2)8CH3 (2h)
按照实施例17的方法,以2.52g(5.49mmol)Nα-Boc-NG-Arg(NO2)-OCH2-(CH2)8CH3为原料,得标题化合物2.15g,为无色固体,产率99%.ESI-MS(m/e):360.7[M+H]+.
实施例25制备HCl·NG-Arg(NO2)-OCH2(CH2)10CH3 (2i)
按照实施例17的方法,以2.0g(4.11mmol)Nα-Boc-NG-Arg(NO2)-OCH2-(CH2)10CH3为原料,得标题化合物1.7g,为无色固体,产率98%.ESI-MS(m/e):388.7[M+H]+.
实施例26制备HCl·NG-Arg(NO2)-OCH2(CH2)12CH3 (2j)
按照实施例17的方法,以1.5g(2.91mmol)Nα-Boc-NG-Arg(NO2)-OCH2-(CH2)12CH3为原料,得标题化合物1.28g,为无色固体,产率97%.ESI-MS(m/e):416.8[M+H]+.
实施例27制备HCl·NG-Arg(NO2)-OCH2(CH2)14CH3 (2k)
按照实施例17的方法,以2.1g(3.87mmol)Nα-Boc-NG-Arg(NO2)-OCH2-(CH2)14CH3为原料,得标题化合物1.81g,为无色固体,产率98%.ESI-MS(m/e):444.5[M+H]+.
实施例28制备HCl·NG-Arg(NO2)-OCH2(CH2)16CH3 (2l)
按照实施例17的方法,以2.0g(3.5mmol)Nα-Boc-NG-Arg(NO2)-OCH2-(CH2)16CH3为原料,得标题化合物1.74g,为无色固体,产率98%.ESI-MS(m/e):472.5[M+H]+.
实施例29制备Boc-Tyr-Arg(NO2)-OBzl
将1.686g(6mmol)Boc-Tyr溶于20ml无水四氢呋喃(THF),冰浴下往里加0.81g(6mmol)N-羟基苯并三唑(HOBt)和1.483g(7.2mmol)二环己基羰二亚胺(DCC)的无水THF溶液。反应混合物冰浴搅拌30分钟,得对应的活泼酯溶液,待用。
将2.2g(7.12mmol)NG-Arg(NO2)-OBzl溶于30ml无水四氢呋喃(THF),然后与上面待用的活泼酯溶液混溶,用N-甲基吗啉(NMM)调pH=8~9。得到的反应混合物室温反应14小时。TLC显示NG Arg(NO2)-OBzl消失。反应混合物减压浓缩至干,残留物用乙酸乙酯溶解,滤除不溶物。滤液依次用5%碳酸氢钠水溶液洗3次,饱和氯化钠水溶液洗3次;5%硫酸氢钾水溶液洗3次,饱和氯化钠水溶液洗3次和饱和碳酸氢钠水溶液洗3次,饱和氯化钠水溶液洗3次。分出的乙酸乙酯层用无水硫酸钠干燥、过滤、滤液32℃减压浓缩,柱层析纯化,得标题化合物2.14g,为无色固体,产率64%.Mp 138℃,ESI-MS(m/z):595[M+Na]+.IR(KBr):3329.59,2949.05,1741.03,1688.89,1649.77,1520.88,1288.35,1173.14,761.23,656.22.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.15(s,1H),8.53(s,1H),8.32-8.30(d,J=6Hz,1H),8.05-7.90(m,2H),7.36(s,5H),7.05-7.02(d,J=9Hz,2H),6.78-6.75(d,J=9Hz,1H),6.66-6.63(d,J=9Hz,2H),5.13(s,2H),4.36-4.35(m,1H),4.13(m,1H),3.16(s,2H),2.83-2.79(m,1H),2.63-2.55(m,1H),1.82-1.78(m,1H),1.70-1.65(m,1H),1.55(m,2H),1.30(s,9H).
实施例30制备Boc-Tyr-NG-Arg(NO2)-NHCH2(CH2)6CH3 (3a)
将2.36g(8.4mmol)Boc-Tyr溶于20ml无水四氢呋喃(THF),冰浴下往里加0.95g(7mmol)N-羟基苯并三唑(HOBt)和1.73g(8.4mmol)二环己基羰二亚胺(DCC)的无水THF溶液。反应混合物冰浴搅拌30分钟,得对应的活泼酯溶液,待用。
将2.6g(7.09mmol)HCl·NG-Arg(NO2)-NHCH2(CH2)6CH3溶于30ml无水四氢呋喃(THF),然后与上面待用的活泼酯溶液混溶,用N-甲基吗啉(NMM)调pH=8~9.得到的反应混合物室温反应14小时。TLC显示HCl·NG-Arg(NO2)-NHCH2-(CH2)6CH3消失。反应混合物减压浓缩至干,残留物用乙酸乙酯溶解,滤除不溶物。滤液依次用5%碳酸氢钠水溶液洗3次,饱和氯化钠水溶液洗3次;5%硫酸氢钾水溶液洗3次,饱和氯化钠水溶液洗3次和饱和碳酸氢钠水溶液洗3次,饱和氯化钠水溶液洗3次。分出的乙酸乙酯层用无水硫酸钠干燥、过滤、滤液32℃减压浓缩,柱层析纯化,得标题化合物2.94g,为无色固体,产率70%.Mp 89℃, ESI-MS(m/z):616.6[M+Na]+.IR(KBr):3316.92,2938.85,1653.84,1527.68,1261.17,571.10.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.15(s,1H),7.86-7.84(d,J=6Hz,2H),7.75(m,1H),7.29-7.26(d,J=9Hz,1H),7.11-7.08(d,J=9Hz,1H),7.03-7.01(d,J=6Hz,2H),6.87-6.85(d,J=6Hz,1H),6.66-6.63(d,J=9Hz,2H),4.25-4.23(m,1H),4.08(m,1H),3.15(s,2H),3.04-3.02(m,2H),2.87-2.83(m,1H),2.63(m,1H),1.65(m,2H),1.48(m,2H),1.39(m,2H),1.31(s,9H),1.23(s,10H),0.86-0.82(t,J=6.0Hz,3H).
实施例31制备Boc-Tyr-NG-Arg(NO2)-NHCH2(CH2)8CH3 (3b)
按照实施例30的方法,1.85g(6.58mmol)Boc-Tyr与1.97g(4.99mmol)HCl·NGArg(NO2)NHCH2(CH2)8CH3反应,得标题化合物1.99g,为无色固体,产率64%.Mp 89℃,ESI-MS(m/z):644.6[M+Na]+.IR(KBr):3317.23,2935.66,2855.93,1650.35,1520.67,1261.36,1165.00,570.50.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.15(s,1H),7.86-7.84(d,J=6Hz,1H),7.75(m,1H),7.43-7.41(d,J=6Hz,1H),7.28-7.26(d,J=6Hz,1H),7.11-7.08(d,J=9Hz,1H),7.03-7.01(d,J=6Hz,1H),6.87-6.84(d,J=9Hz,1H),6.65-6.63(d,J=6Hz,2H),4.24(s,1H),4.07(m,1H),3.15(m,2H),3.04-3.02(m,3H),2.86-2.83(m,1H),2.67-2.59(m,1H),1.65(m,2H),1.48(m,2H),1.37(m,2H),1.31(s,9H),1.23(s,14H),0.85-0.81(t,J=6.0Hz,3H).
实施例32制备Boc-Tyr-NG-Arg(NO2)-NHCH2(CH2)10CH3(3c)
按照实施例30的方法,3.0g(10.7mmol)Boc-Tyr与2.98g(7.05mmol)HCl·NG-Arg(NO2)-NHCH2(CH2)10CH3反应,得标题化合物2.0g,为无色固体,产率44%.Mp 92℃,ESI-MS(m/z):672.2.[M+Na]+.IR(KBr):3316.06,2933.83,2855.41,1650.59,1527.48,1261.58,1164.80,572.14.1H-N-MR(300MHz,DMSO-d6):δ/ppm=9.19(s,1H),8.55(s,1H),7.91-7.88(d,J=9Hz,2H),7.79(m,2H),7.04-7.01(d,J=9Hz,2H),6.91-6.88(d,J=9Hz,1H),6.65-6.62(d,J=9Hz,2H),4.40-4.14-4.04(m,1H),3.14(s,2H),3.02(m,2H),2.87-2.81(dd,J=3Hz,J=15Hz,1H),2.66-2.57(dd,J=12Hz,J=15Hz,1H),1.65(m,2H),1.47(m,2H),1.36(m,2H),1.30(s,9H),1.22(s,22H),0.86-0.82(t,J=6.0Hz,3H).
实施例33制备Boc-Tyr-NG-Arg(NO2)-NHCH2(CH2)12CH3(3d)
按照实施例30的方法,2.48g(8.83mmol)Boc-Tyr与3.33g(7.39mmol)HCl·NG-Arg(NO2)-NHCH2(CH2)12CH3反应,得标题化合物1.6g,为无色固体,产率32%.Mp 94℃,ESI-MS(m/z):700.3[M+Na]+.IR(KBr):3317.37,2931.77,2854.99,1649.60,1526.73,1261.21,573.55.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.19(s,1H),7.90-7.87(d,J=9Hz,1H),7.79(m,1H),7.03-7.00(d,J=9Hz,2H),6.92-6.89(d,J=9Hz,1H),6.65-6.62(d,J=9Hz,2H),4.22(m,1H),4.06-4.03(m,1H),3.14(m,2H),3.02-3.01(m,2H),2.86-2.81(dd,J=3Hz,J=12Hz,1H),2.65-2.57(m,1H),1.64(m,2H),1.47(m,2H),1.36(m,2H),1.30(s,9H),1.22(s,18H),0.86-0.82(t,J=6.0Hz,3H).
实施例34制备Boc-Tyr-NG-Arg(NO2)-NHCH2(CH2)14CH3(3e)
按照实施例30的方法,2.48g(8.83mmol)Boc-Tyr与3.5g(7.31mmol)HCl·NG-Arg(NO2)-NHCH2(CH2)14CH3反应,得标题化合物2.4g,为无色固体,产率47%.Mp 94℃,ESI-MS(m/z):728.2[M+Na]+.IR(KBr):3316.91,2930.16,2855.20,1648.65,1526.68,1261.70,1165.91,573.19.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.15(s,1H),8.52(s,1H),7.86-7.84(d,J=6Hz,1H),7.76(m,1H),7.03-7.00(d,J=9Hz,2H),6.87-6.84(d,J=9Hz,1H),6.65-6.62(d,J=9Hz,2H),4.23(m,1H),4.07-4.04(m,1H),3.14(s,2H),3.02(m,2H),2.87-2.82(dd,J=6Hz,J=12Hz,1H),2.66-2.58(m,1H),1.65(m,2H),1.47(m,2H),1.36(m,2H),1.30(s,9H),1.22(s,26H),0.86-0.82(t,J=6.0Hz,3H).
实施例35制备Boc-Tyr-NG-Arg(NO2)-NHCH2(CH2)16CH3(3f)
按照实施例30的方法,3.0g(10.7mmol)Boc-Tyr与3.84g(7.58mmol)HCl·NG-Arg(NO2)-NHCH2(CH2)16CH3反应,得标题化合物1.6g,为无色固体,产率29%.Mp 94℃,ESI-MS(m/z):756.6[M+Na]+.IR(KBr):3316.19.2928.28,2853.24,1650.90,1522.56,1259.79.1166.06,579.27.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.15(s,1H),8.52(s,1H),7.87-7.84(d,J=9Hz,1H),7.76(m,1H),7.42-7.40(1H),7.28-7.25(1H),7.03-7.00(d,J=9Hz,2H),6.86-6.84(d,J=6Hz,1H),6.65-6.62(d,J=9Hz,2H),4.24(m,1H),4.08-4.01(m,1H),3.16-3.15(m,2H),3.02(m,2H),2.88-2.82(dd,J=3Hz,J=15Hz,1H),2.67-2.59(m,1H),1.65(m,2H),1.48(m,2H),1.36(m,2H),1.30(s,9H),1.22(s,30H),0.86-0.82(t,J=6.0Hz,3H).
实施例36制备Boc-Tyr-NG-Arg(NO2)-OCH2(CH2)6CH3 (3g)
按照实施例30的方法,1.58g(5.62mmol)Boc-Tyr与1.73g(4.71mmol)HCl·NG-Arg(NO2)-OCH2(CH2)6CH3反应,得标题化合物0.85g,为无色固体,产率30%,Mp 52℃,ESI-MS(m/z):617.6[M+Na]+.IR(KBr):3333.97,2941.57,2358.30,1675.38,1516.92,1260.81,666.94.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.17(s,1H),8.54(s,1H),8.29-8.27(d,J=6Hz,2H),7.31-7.29(m,1H),7.07-7.04(d,J=9Hz,2H),6.81-6.78(d,J=9Hz,1H),6.66-6.63(d,J=9Hz,2H),4.28-4.22(m,1H),4.15-4.07(m,1H),4.05-3.97(t,J=12Hz,2H),3.16(m,2H),2.87-2.81(dd,J=3Hz J=15Hz,1H),2.64-2.57(dd,J=9Hz,J=12Hz,1H),1.87-1.62(m,2H),1.57-1.53(m,4H),1.30(s,9H),1.21(s,10H),0.86-0.82(t,J=6.0Hz,3H).
实施例37制备Boc-Tyr-NG-Arg(NO2)-OCH2(CH2)8CH3 (3h)
按照实施例30的方法,1.36g(4.84mmol)Boc-Tyr与1.6g(4.04mmol)HCl·NG-Arg(NO2)-OCH2(CH2)8CH3反应,得标题化合物0.8g,为无色固体,产率32%.Mp 71℃,ESI-MS(m/z):645.6[M+Na]+.IR(KBr):3326.44,2936.49,2856.02,2357.75,1691.97,1521.02,1287.85,653.87.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.18(s,1H),8.54(s,1H),8.30-8.27(d,J=9Hz,1H),7.90(s,1H),7.07-7.04(d,J=9Hz,2H),6.82-6.79(d,J=9Hz,1H),6.66-6.63(d,J=9Hz,2H),4.28-4.23(m,1H),4.13-4.03(m,1H),4.00-3.94(t,J=9Hz,2H),3.16(m,2H),2.86-2.81(dd,J=3Hz,J=12Hz,1H),2.64-2.56(dd,J=12Hz,J=15Hz,1H),1.75-1.73(m,2H),1.66-1.61(m,2H),1.56-1.52(t,J=6Hz,2H),1.30(s,9H),1.21(s,14H),0.86-0.82(t,J=6.0Hz,3H).
实施例38制备Boc-Tyr-NG-Arg(NO2)-OCH2(CH2)10CH3 (3i)
按照实施例30的方法,1.39g(4.95mmol)Boc-Tyr与1.7g(4.01mmol)HCl·NG-Arg(NO2)-OCH2(CH2)10CH3反应,得标题化合物0.9g,为无色固体,产率35%.Mp 61℃,ESI-MS(m/z):673.7[M+Na]+.IR(KBr):3329.90,2935.98,2855.48,2343.29,1665.31,1518.00,1263.54,678.93.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.18(s,1H),8.29-8.26(d,J=9Hz,1H),7.31-7.28(d,J=9Hz,1H),7.07-7.04(d,J=9Hz,2H),6.94(s,1H),6.81-6.78(d,J=9Hz,1H),6.66-6.63(d,J=9Hz,2H),4.28-4.24(m,1H),4.15-4.07(m,1H),4.04-4.01(t,J=9Hz,2H),3.16(m,2H),2.8-2.81(dd,J=3Hz,J=12Hz,1H),2.64-2.56(m,1H),1.75-1.71(m,2H),1.66-1.61(m,2H),1.56(m,4H),1.30(s,9H),1.21(s,18H),0.86-0.82(t,J=6.0Hz,3H).
实施例39制备Boc-Tyr-NG-Arg(NO2)-OCH2(CH2)12CH3 (3j)
按照实施例30的方法,1.27g(4.52mmol)Boc-Tyr与1.28g(2.83mmol)HCl·NG-Arg(NO2)-OCH2(CH2)12CH3反应,得标题化合物0.7g,为无色固体,产率36%.Mp 61℃,ESI-MS(m/z):702.2[M+Na]+.IR(KBr):3330.88,2934.28,2856.88,2344.26,1665.64,1519.66,1261.88,674.55.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.16(s,1H),8.54(s,1H),8.29-8.26(d,J=9Hz,1H),7.92-7.90(m,1H),7.07-7.04(d,J=9Hz,2H),6.80-6.77(d,J=9Hz,1H),6.66-6.63(d,J=9Hz,2H),4.28-4.27(m,1H),4.12-4.09(m,1H),4.05-4.01(t,J=6Hz,2H),3.16(m,2H),2.87-2.81(dd,J=3Hz J=15Hz 1H),2.61-2.51(dd,J=12Hz,J=18Hz,1H),1.76-1.74(m,2H),1.70-1.62(m,2H),1.56-1.52(t,J=6Hz,2H),1.30(s,9H),1.23-1.21(m,22H),0.87-0.83(t,J=6.0Hz,3H).
实施例40制备Boc-Tyr-NG-Arg(NO2)-OCH2(CH2)14CH3(3k)
按照实施例30的方法,1.43g(5.08mmol)Boc-Tyr与1.81g(3.77mmol)HCl·NG-Arg(NO2)-OCH2(CH2)14CH3反应,得标题化合物1.2g,为无色固体,产率45%.Mp 65℃,ESI-MS(m/z):729.5[M+Na]+.IR(KBr):3325.38,2925.23,2854.50,2344.11,1673.64,1516.47,1262.22,1166.50,686.41.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.16(s,1H),8.53(s,1H),8.29-8.26(d,J=9Hz,1H),7.31-7.28(d,J=9Hz,1H),7.07-7.04(d,J=9Hz,2H),6.80-6.77(d,J=9Hz,1H),6.66-6.63(d,J=9Hz,2H),4.29-4.25(m,1H),4.12-4.08(dd,J=3Hz,J=9Hz,1H),4.05-4.01(t,J=6Hz,2H),3.16(m,2H),2.86-2.81(dd,J=3Hz,J=12Hz,1H),2.64-2.56(dd,J=9Hz,J=12Hz,1H),1.75-1.74(m,2H),1.69-1.62(m,2H),1.56-1.52(t,J=6Hz,2H),1.30(s,9H),1.23-1.21(m,26H),0.87-0.83(t,J=6.0Hz,3H).
实施例41制备Boc-Tyr-NG-Arg(NO2)-OCH2(CH2)16CH3 (3l)
按照实施例30的方法,1.28g(4.56mmol)Boc-Tyr与1.77g(3.48mmol)HCl·NG-Arg(NO2)-OCH2(CH2)16CH3反应,得标题化合物1.1g,为无色固体,产率43%.Mp 74℃,ESI-MS(m/z):757.5[M+Na]+.IR(KBr):3319.66,2924.78,2853.44,2344.15,1652.06,1520.03,1254.32,1171.41,659.46.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.26(s,1H),8.53(s,1H),8.28-8.26(d,J=6Hz,1H),7.31-7.28(d,J=9Hz,1H),7.07-7.04(d,J=9Hz,2H),6.93-6.86(m,1H),6.79-6.76(d,J=9Hz,1H),6.66-6.63(d,J=9Hz,2H),4.38(s,1H),4.27(m,1H),4.09-4.01(t,J=12Hz,2H),3.16(s,2H),2.86-2.82(m,1H),2.65-2.57(t,J=12Hz,1H),1.76-1.74(m,1H),1.67-1.62(m,1H),1.56-1.52(m,4H),1.30(s,9H),1.23-1.21(m,30H),0.87-0.83(t,J=6.0Hz,3H).
实施例42制备Tyr-Arg
将900mg(1.573mmol)Boc-Tyr-NG-Arg(NO2)-OBzl用15ml CH3OH溶解,再加入270mg Pd/C(5%),减压排出反应瓶中的空气,通入氢气置换,反复置换三次后,通氢气室温搅拌反应,直至TLC显示Boc-Tyr-NG-Arg(NO2)-OBzl消失。停止反应,滤除Pd/C,滤液减压浓缩至干,得Boc-Tyr-Arg 673mg,为无色固体。
取Boc-Tyr-Arg 673mg溶于适量乙酸乙酯,加入约15ml 4N盐酸乙酸乙酯液,室温搅拌3小时,TLC显示原料点消失。反应混合液在室温下减压浓缩,残留物再用乙酸乙酯溶解并室温下浓缩,如此反复5次;然后向浓缩物中加入乙醚,室温下减压浓缩,如此反复5次,直至除净游离的氯化氢,得标题化合物480mg,为无色固体,产率91%.Mp 120-121℃,ESI-MS(m/z):338.2[M+Na]+.IR(KBr):3169.98,1665.50,1514.29,1419.94,1234.60,688.17.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.42-9.39(m,1H),9.21-9.19(d,J=6Hz,1H),9.09-9.07(d,J=6Hz,1H),8.26(m,2H),7.97(s,1H),7.59-7.53(t,J=9Hz,1H),7.42-7.36(t,J=9Hz,1H),7.25-7.22(d,J=9Hz,1H),7.14-7.12(d,J=6Hz,2H),6.72-6.69(d,J=9Hz,2H),4.26-4.21(dd,J=6Hz,J=9Hz,1H),4.10(m,1H),3.14-3.12(m,2H),3.07-3.06(m,1H),2.88-2.80(dd,J=9Hz,J=15Hz,1H),1.80-1.78(m,1H),1.72-1.66(m,1H),1.59-1.57(m,2H).
实施例43制备Tyr-Arg-NHCH2(CH2)6CH3 (5a)
按照实施例42的方法,以800mg(1.35mmol)Boc-Tyr-NG-Arg(NO2)-NHCH2(CH2)6CH3为原料,得标题化合物560mg,为无色固体,产率93%.Mp 122-123℃,ESI-MS(m/z):449.5[M+H]+.IR(KBr):3255.66,2936.75,1656.00,1533.13,1235.53,570.88.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.32(s,1H),8.87-8.85(d,J=6Hz,1H),8.33-8.31(d,J=6Hz,1H),8.28(m,1H),7.99(m,1H),7.50(s,2H),7.26(s,1H),7.07-7.04(d,J=9Hz,2H),6.70-6.67(d,J=9Hz,2H),4.27-4.21(Q,J=6Hz,1H),4.06(s,1H),3.15-3.12(m,2H),3.06-3.00(m,3H),2.88-2.82(dd,J=6Hz,J=12Hz 1H),1.73(m,2H),1.58-1.45(m,2H),1.39(m,2H),1.23(s,10H),0.86-0.82(t,J=6.0Hz,3H).
实施例44制备Tyr-Arg-NHCH2(CH2)8CH3 (5b)
按照实施例42的方法,以700mg(1.13mmol)Boc-Tyr-NG-Arg(NO2)-NHCH2(CH2)8CH3为原料,得标题化合物530mg,为无色固体,产率99%.Mp 125-126℃,ESI-MS(m/z):477.5[M+H]+.IR(KBr):3196.55,2935.38,1656.57,1533.25,1238.09,569.57.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.39(s,1H),8.88-8.86(d,J=6Hz,2H),8.31-8.30(d,J=3Hz,1H),8.27-8.25(d,J=6Hz,1H),8.01-7.96(m,2H),7.49(s,2H),7.07-7.04(d,J=9Hz,2H),6.70-6.67(d,J=9Hz,2H),4.28-4.24(m,1H),4.06-4.05(m,1H),3.12(m,2H),3.05-3.01(m,3H),2.87-2.80(dd,J=6Hz,J=15Hz 1H),1.72-1.63(m,1H),1.58-1.57(m,1H),1.51-1.45(m,2H),1.22(s,14H),0.86-0.82(t,J=6.0Hz,3H).
实施例45制备Tyr-Arg-NHCH2(CH2)10CH3(5c)
按照实施例42的方法,以930mg(1.43mmol)Boc-Tyr-NG-Arg(NO2)-NHCH2(CH2)10CH3为原料,得标题化合物710mg,为无色固体,产率98%.Mp 129-130℃,ESI-MS(m/z):505.5[M+H]+.IR(KBr):3413.08,2936.07,1650.61,1241.43,1109.47,574.11.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.31(s,1H),8.88-8.86(d,J=6Hz,1H),8.33-8.31(d,J=6Hz,1H),8.28(m,1H),8.00(s,2H),7.51(s,1H),7.33-7.24(m,2H),7.07-7.04(d,J=9Hz,2H),6.70-6.67(d,J=9Hz,2H),4.42-4.40(m,1H),4.06(s,1H),3.12(m,2H),3.06-3.00(Q,J=6Hz,3H),2.88-2.81(dd,J=6Hz,J=15Hz,1H),1.68-1.64(m,2H),1.59-1.48(m,2H),1.38(m,2H),1.22(s,18H),0.87-0.83(t,J=6.0Hz,3H).
实施例46制备Tyr-Arg-NHCH2(CH2)12CH3 (5d)
按照实施例42的方法,以806mg(1.19mmol)Boc-Tyr-NG-Arg(NO2)-NHCH2(CH2)12CH3为原料,得标题化合物618mg,为无色固体,产率79%.Mp 138-139℃,ESI-MS(m/z):533.5[M+H]+.IR(KBr):3368.55,3200.21,2920.03,1658.31,1472.16,1229.38,680.09.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.38(s,1H),8.87-8.85(d,J=6Hz,1H),8.22(s,2H),8.00-7.98(d,J=6Hz,2H),7.39-7.11(m,3H),7.07-7.05(d,J=6Hz,2H),6.70-6.67(d,J=9Hz,2H),4.26-4.21(m,1H),4.08-4.04(t,J=6Hz,2H),3.14-3.12(m,2H),3.06-3.00(Q,J=6Hz,3H),2.88-2.82(dd,J=6Hz,J=12Hz 1H),1.69(m,1H),1.59-1.49(m,3H),1.39(m,2H),1.23(s,22H),0.87-0.83(t,J=6.0Hz,3H).
实施例47制备Tyr-Arg-NHCH2(CH2)14CH3 (5e)
按照实施例42的方法,以400mg(0.57mmol)Boc-Tyr-NG-Arg(NO2)-NHCH2(CH2)14CH3为原料,得标题化合物320mg,为无色固体,产率94%.Mp 142-143℃,ESI-MS(m/z):561.8[M+H]+.IR(KBr):3222.60,2926.35,1650.41,1551.58,1474.24,1227.14,681.87.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.40(s,1H),8.89-8.87(d,J=6Hz,1H),8.25(s,3H),8.01-7.99(d,J=6Hz,2H),7.46-7.44(m,1H),7.07-7.04(d,J=9Hz,3H),6.70-6.67(d,J=9Hz,2H),4.25-4.21(m,1H),4.06(m,1H),3.14-3.12(m,2H),3.04-3.02(m,3H),2.89-2.82(dd,J=6Hz,J=9Hz 1H),1.69(m,2H),1.59-1.49(m,2H),1.39(m,2H),1.22(s,26H),0.87-0.83(t,J=6.0Hz,3H).
实施例48制备Tyr-Arg-NHCH2(CH2)16CH3(5f)
按照实施例42的方法,以500mg(0.69mmol)Boc-Tyr-NG-Arg(NO2)-NHCH2(CH2)16CH3为原料,得标题化合物300mg,为无色固体,产率70%.Mp 145-146℃,ESI-MS(m/z):589.5[M+H]+.IR(KBr):3292.18,2927.61,1657.73,1515.64,1240.28,571.51.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.36(s,1H),8.87-8.85(d,J=6Hz,1H),8.33-8.31(d,J=6Hz,1H),8.28-8.27(d,J=3Hz,1H),7.98(m,2H),7.47-7.16(m,3H),7.07-7.04(d,J=9Hz,2H),6.70-6.65(d,J=9Hz,2H),4.23(m,1H),4.06(s,1H),3.12(m,2H),3.04-3.00(m,3H),2.89-2.81(dd,J=9Hz,J=15Hz 1H),1.68-1.64(m,2H),1.59-1.49(m,2H),1.38(m,2H),1.22(s,30H),0.86-0.82(t,J=6.0Hz,3H).
实施例49制备Tyr-Arg-OCH2(CH2)6CH3 (5g)
按照实施例42的方法,以600mg(1.01mmol)Boc-Tyr-NG-Arg(NO2)-OCH2(CH2)6CH3为原料,得标题化合物380mg,为无色固体,产率77%.Mp 106-107℃,ESI-MS(m/z):449.3[M+H]+.IR(KBr):3187.11,2937.87,1662.85,1513.63,1232.09,573.29.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.32(s,1H),9.28-9.26(d,J=6Hz,1H),8.32-8.27(m,3H),8.02(s,1H),7.51-7.41(m,1H),7.14-7.11(d,J=9Hz,2H),7.02-6.93(m,1H),6.72-6.69(d,J=9Hz,2H),4.28-4.27(m,1H),4.07-4.02(m,3H),3.13-3.11(m,2H),3.07-3.05(m,1H),2.86-2.78(dd,J=9Hz,J=12Hz,1H),1.80-1.77(m,2H),1.72-1.67(m,2H),1.57-1.56(m,2H),1.22(s,10H),0.85-0.81(t,J=6.0Hz,3H).
实施例50制备Tyr-Arg-OCH2(CH2)8CH3(5h)
按照实施例42的方法,以600mg(0.96mmol)Boc-Tyr-NG-Arg(NO2)-OCH2(CH2)8CH3为原料,得标题化合物420mg,为无色固体,产率85%.Mp 107-108℃,ESI-MS(m/z):478.4[M+H]+.IR(KBr):3442.53,3221.27,3093.95,2941.87,1656.80,1221.75.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.45(s,1H),9.27-9.25(d,J=6Hz,1H),8.30-8.29(d,J=3Hz,1H),8.25(s,1H),8.00(m,1H),7.58-7.55(m,1H),7.44-7.41(m,1H),7.14-7.11(d,J=9Hz,2H),6.99-6.93(t,J=9Hz,1H),6.72-6.69(d,J=9Hz,2H),4.30-4.26(m,1H),4.19-4.14(m,3H),3.13-3.10(m,2H),3.07-3.05(m,1H),2.85-2.77(dd,J=9Hz,J=15Hz,1H),1.81-1.78(m,2H),1.74-1.67(m,2H),1.62-1.56(m,2H),1.21(s,14H),0.86-0.82(t,J=6.0Hz,3H).
实施例51制备Tyr-Arg-OCH2(CH2)10CH3(5i)
按照实施例42的方法,以850mg(1.31mmol)Boc-Tyr-NG-Arg(NO2)-OCH2(CH2)10CH3为原料,得标题化合物550mg,为无色固体,产率78%.Mp 111-112℃,ESI-MS(m/z):506.5[M+H]+.IR(KBr):3188.23,2932.80,1666.24,1514.82,1231.76,572.08.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.45(s,1H),9.28-9.25(d,J=9Hz,2H),8.23(s,3H),8.05-8.02(d,J=3Hz,1H),7.14-7.11(d,J=9Hz,3H),6.72-6.69(d,J=9Hz,2H),4.31-4.25(q,J=6Hz,1H),4.06-4.02(t,J=6Hz,3H),3.16-3.10(q,J=6Hz,2H),3.07-3.03(m,1H),2.86-2.78(dd,J=9Hz,J=15Hz,1H),1.86-1.70(m,2H),1.62-1.54(m,4H),1.22(s,18H),0.87-0.83(t,J=6.0Hz,3H).
实施例52制备Tyr-Arg-OCH2(CH2)12CH3 (5j)
按照实施例42的方法,以600mg(0.88mmol)Boc-Tyr-NG-Arg(NO2)-OCH2(CH2)12CH3为原料,得标题化合物360mg,为无色固体,产率72%.Mp 136-137℃,ESI-MS(m/z):534.5[M+H]+.IR(KBr):3180.27,2928.00.1661.37,1510.73,1228.78,669.56.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.44(s,1H),9.27-9.25(d,J=6Hz,2H),8.30-8.28(d,J=6Hz,1H),8.25(m,1H),8.00(m,1H),7.44-7.41(m,2H),7.13-7.11(d,J=6Hz,2H),6.72-6.69(d,J=9Hz,2H),4.29-4.25(m,1H),4.07-4.02(m,3H),3.12-3.10(m,2H),3.08-3.05(m,1H),2.84-2.77(dd,J=9Hz,J=15Hz,1H),1.84-1.76(m,2H),1.74-1.66(m,2H),1.62-1.55(m,2H),1.22(s,22H),0.87-0.83(t,J=6.0Hz,3H).
实施例53制备Tyr-Arg-OCH2(CH2)14CH3 (5k)
按照实施例42的方法,以750mg(1.06mmol)Boc-Tyr-NG-Arg(NO2)-OCH2(CH2)14CH3为原料,得标题化合物510mg,为无色固体,产率81%.Mp 140-141℃,ESI-MS(m/z):562.5[M+H]+.IR(KBr):3185.75,2929.25,1661.37,1512.85,1232.12,832.85,653.79.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.44(s,1H),9.26-9.24(d,J=6Hz,2H),8.30-8.28(d,J=6Hz,1H),8.25(s,1H),7.99(m,1H),7.50(m,1H),7.13-7.11(d,J=6Hz,2H),6.99-6.96(d,J=9Hz,1H),6.72-6.69(d,J=9Hz,2H),4.29-4.28(m,1H),4.07-4.02(m,3H),3.13-3.10(m,2H),3.07-3.02(m,1H),2.85-2.77(dd,J=9Hz J=15Hz,1H),1.78-1.74(m,2H),1.72-1.67(m,2H),1.62-1.55(m,2H),1.22(s,26H),0.87-0.83(t,J=6.0Hz,3H).
实施例54制备Tyr-Arg-OCH2(CH2)16CH3 (5l)
按照实施例42的方法,以700mg(0.96mmol)Boc-Tyr-NG-Arg(NO2)-OCH2(CH2)16CH3为原料,得标题化合物500mg,为无色固体,产率83%.Mp 145-146℃,ESI-MS(m/z):590.6[M+H]+.IR(KBr):3189.43,2929.08,1661.01,1514.60,1232.87.833.09,575.19.1H-NMR(300MHz,DMSO-d6):δ/ppm=9.44(s,1H),9.27-9.25(d,J=6Hz,2H),8.30-8.29(d,J=3Hz,1H),8.26(m,1H),8.00(m,1H),7.51(m,1H),7.14-7.11(d,J=9Hz,2H),7.02-6.99(d,J=9Hz,1H),6.72-6.69(d,J=9Hz,2H),4.29-4.25(m,1H),4.06-4.02(m,3H),3.15-3.10(m,2H),3.07-3.00(m,1H),2.85-2.77(dd,J=9Hz,J=15Hz,1H),1.78-1.74(m,2H),1.72-1.65(m,2H),1.62-1.53(m,2H),1.23(s,30H),0.87-0.83(t,J=6.0Hz,3H).
试验例1本发明的化合物5a-l的镇痛活性实验
1)受试化合物:本发明的化合物5a-l
阳性对照品:阿司匹林;
2)实验动物:ICR小鼠,雄性,体重20±2g;每12只小鼠一组,空白及阳性对照各一组。
3)剂量设置:阿司匹林165μmol/kg,Tyr-Arg 100μmol/kg,5a-l 10μmol/kg,均单次灌胃。
药物配制:生理盐水(NS)溶解。
4)给药方案:0.2ml溶液/鼠。
5)动物模型
ICR雄性小鼠使用前静息1天,操作间保持室内温度22℃,每组小鼠12只。基础痛阈的测定是在开始时,先测三次,每次间隔5分钟,取其平均值为基础痛阈。每次试验设生理盐水组为平行对照。
6)痛阈提高率的测定
灌胃后每30min测定一次痛阈值,共测定6组180min。痛阈改变以下列公式表示:痛阈提高%=[(给药后痛阈-基础痛阈)/基础痛阈]×100%。
7)统计方法
本实验数据统计均采用t检验和方差分析,以X±SD表示。
8)实验结果
受试化合物热辐射甩尾实验痛阈提高率结果如表1所示。在10μmol/kg剂量下受试化合物经灌胃给药后对疼痛有显著的抑制作用。
表1化合物5a-l的镇痛作用
Aspirin剂量:165μmol/kg,Tyr-Arg剂量:100μmol/kg,5a-l剂量:10μmol/kg,a)与NS组比较P<0.01;b)与NS组比较P<0.05;c)与NS组比较P<0.01,与Tyr-Arg组比较P<0.01;d)与NS组比较P<0.01,与Tyr-Arg组比较P<0.05;e)与NS组比较P<0.01,与Tyr-Arg组比较P<0.01,与Aspirin比较P<0.01;f)与NS组比较P<0.01,与Tyr-Arg组比较P<0.01,与Aspirin比较P<0.05;g)与NS组比较P<0.01,与Tyr-Arg组比较P<0.05,与Aspirin比较P<0.01;h)与NS组比较P<0.01,与Tyr-Arg组比较P<0.05,与Aspirin比较P<0.05;i)与NS组比较P<0.01,与Aspirin比较P<0.01;j)与NS组比较P<0.01,与Aspirin比较P<0.05
试验例2本发明化合物5a,5h的镇痛活性剂量和效应的关系
试验方法同试验例1,受试化合物为5a,5h;将受试化合物分别按10μmol/kg,1μmol/kg和0.1μmol/kg剂量,均采用灌胃单次给药。试验结果见表2。实验结果表明,本发明化合物5a,5h的镇痛活性呈现剂量依赖关系。
表2不同剂量5a,5h的镇痛活性
n=12;受试化合物5a,5h的剂量分别为H=10μmol/kg,M=1μmol/kg和L=0.1μmol/kg.a)与NS组比较p<0.01;b)与NS组比较p<0.05;c)与NS组比较p<0.01,与M组比较p<0.01;d)与NS组比较p<0.01,与M组比较p<0.05,e)与NS组比较p<0.05,与M组比较p<0.01
实验例3本发明化合物5a-l的抗炎活性评价
化合物5a-l或阿司匹林用生理盐水配成溶液,雄性ICR小鼠(20±2g)随机分为空白对照组、阿司匹林组、Tyr-Arg及5a-l组,每组12只小鼠。分别灌胃给予化合物5a-l(剂量为10μmol/kg)或阿司匹林(剂量为165μmol/kg)溶液0.2ml/只,给药30分钟后,往小白鼠的左耳外廓涂0.03ml二甲苯。2小时后将小白鼠颈椎脱臼处死、两耳用直径7mm的打孔器取圆形耳片、称重、把两圆耳片的重量差作为肿胀度。结果列入表3。实验结果表明,本发明的化合物5a-l具有明确的抗炎活性
表3化合物5a-l对小鼠耳肿胀的抑制作用
阿司匹林剂量为165μmol/kg,Tyr-Arg剂量为100μmol/kg;5a-l剂量为10μmol/kg;n=12,a)与NS组比较P<0.01;b)与NS组比较P<0.0,1与Aspirin比较P<0.05;c)与NS组比较P<0.01,与Tyr-Arg比较p<0.05;d)与NS组比较P<0.01,与Aspirin比较P<0.05,与Tyr-Arg比较p<0.05
实验例4不同剂量5a,5h的抗炎活性评价
按照实验例3的方法,5a,5h均按10μmol/kg、1μmol/kg、0.1μmol/kg三种剂量给雄性ICR小鼠灌胃,两圆耳片的重量差列入表4。结果表明本发明的化合物5a,5h的抗炎活性具有剂量依赖性。
表4不同剂量5a,5h对小鼠耳肿胀的抑制作用
a)与NS组比较p<0.01;b)与NS组比较p<0.01,与1μmol/kg组比较p<0.01;c)与NS组比较p<0.01,与1μmol/kg组比较p<0.05
以上所述的实施例仅仅是对本发明的优选实施方式进行描述,并非对本发明的范围进行限定,在不脱离本发明设计精神的前提下,本领域普通技术人员对本发明的技术方案作出的各种变形和改进,均应落入本发明权利要求书确定的保护范围内。
Claims (5)
1.一种通式为I的化合物:
Tyr-Arg-XCH2(CH2)nCH3
I
式中,X为NH或O;
式中,n为6、8、10、12、14或16。
2.一种制备权利要求1所述的化合物的方法,其特征在于包括以下步骤:
(1)浓硫酸、浓硝酸条件下,Arg硝基化得NG-Arg(NO2);
(2)弱碱性水二氧六环条件下,NG-Arg(NO2)与(Boc)2O反应制得Nα-Boc-NG-Arg(NO2);
(3)在DCC和HOBt存在下,Nα-Boc-NG-Arg(NO2)在无水THF中与脂肪胺或脂肪醇缩合成Nα-Boc-NG-Arg(NO2)-XCH2(CH2)nCH3;
(4)在含氯化氢的乙酸乙酯中,Nα-Boc-NG-Arg(NO2)-XCH2(CH2)nCH3脱Boc保护,得NG-Arg(NO2)-XCH2(CH2)nCH3;
(5)在DCC和HOBt存在下,Boc-Tyr在无水THF中与NG-Arg(NO2)-XCH2-(CH2)nCH3缩合,制备得到Boc-Tyr-NG-Arg(NO2)-XCH2(CH2)nCH3;
(6)H2/Pd条件下Boc-Tyr-NG-Arg(NO2)-XCH2(CH2)nCH3脱-NO2保护,得Boc-Tyr-Arg-XCH2(CH2)nCH3;
(7)在含氯化氢的乙酸乙酯中,Boc-Tyr-Arg-XCH2(CH2)nCH3脱Boc保护,即得Tyr-Arg-XCH2(CH2)nCH3。
3.根据权利要求2所述的方法,其特征在于:所述步骤(3)中所述的脂肪胺为正八胺、正十胺、正十二胺、正十四胺、正十六胺或正十八胺;所述脂肪醇为正八醇、正十醇、正十二醇、正十四醇、正十六醇或正十八醇 。
4.根据权利要求2所述的方法,其特征在于:所述Arg和Tyr均为L构型氨基酸。
5.权利要求1所述的化合物在制备镇痛和抗炎药物中的用途。
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