新霉胺-咔啉羧酸缀合物及其制备方法和在医学中的用途的制作方法

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专利名称::新霉胺-咔啉羧酸缀合物及其制备方法和在医学中的用途的制作方法
技术领域
:本发明涉及一类具有抑菌活性的缀合物,尤其涉及由新霉胺和咔啉羧酸缀合而成的新霉胺-咔啉羧酸缀合物及其制备方法,本发明还涉及新霉胺-咔啉羧酸缀合物在抗菌、抗艾滋病病毒中的用途,属于药物化学领域。
背景技术
:氨基糖苷类抗生素是在临床广泛使用的一类广谱抗生素,作用强,主要用于革兰阳性和阴性杆菌所致的全身感染。由于该类药物为多氨基多羟基,是一类高极性化合物,口服无效,只能成盐后注射给药。此外,该类药物具有耳毒性和肾毒性。并且由于在临床上使用时间已久和使用不当,已经出现了严重的耐药性。对于其作用机理研究发现,该类药物以rRNA,tRNA以及mRNA等多种RNA为靶,以氢键、静电、堆叠嵌插等作用于细菌或病毒rRNA,使得rRNA在由mRNA翻译成蛋白质的过程中产生错误,或者对翻译的进程产生影响,进而影响到细菌或病毒细胞的正常生理生化功能,造成细菌或病毒的繁殖受到抑制(Moazed,D.;Noller,H.F.Wature1987,327'389—394;Fourmy,D.;Recht,M.I.;Blanchard,S.C.;Puglisi,J.D.Science1996,274;Carter,A.P.;Clemons,W.M.;Brodersen,D.E.;Morgan-Warren,R._J.;Wimberly,B.T.;Ramakrish醒,V.淑ure2000,340-348.)。尤其是氨基糖苷类抗生素可以作用在HIV-1基因表达过程的反式激活环节,阻断转录的反式激活子Tat(transactivitoroftranscription)蛋白和TARRNA(transactivitorresponseregionRNA)之间的结合,从而可以阻断HIV-1基因的表达,因此氨基糖苷类抗生素重新引起世界范围内的研究兴趣。新霉胺单元是在天然的氨基糖苷类抗生素中广为存在的最基本的药效结构单元,以新霉胺为先导化合物开展了大量的工作,但是单纯从提高化合物与受体的氢键、静电结合能力入手,获得的化合物相对于母体新霉素或新霉胺,其活性并没有实质性提高。咔啉羧酸为存在于我国南方食用植物蠤头中的成分,无毒,其芳构化衍生物具有优秀的抗HIV活性和抗菌活性(Ishida,J.;Wang,H.K.;Bastow,K.F.;Hu,C.Q.;Lee,K.fiioorg.ifed.Oie瓜Lett.1999,9,3319-3324.)。因此,我们将两者的结构片段相缀合,期望能起到协同增效的作用。
发明内容本发明目的之一是提供一类新霉胺-咔啉羧酸缀合物实体;本发明目的之二是提供一类制备上述新霉胺-咔啉羧酸缀合物的方法;本发明目的之三是将上述新霉胺-咔啉羧酸缀合物应用于制备成抗菌药物、抗艾滋病药物;本发明上述目的是通过以下技术方案来实现的新霉胺-咔啉羧酸缀合物,其结构式为通式i或通式n所示通式I通式II其中,R选自氢或CH30;R'选自氢、甲基或CH2CH(0CH3)2;X选自CONH,NHCO,NH,CO或氧;Y为氧,CH2,NH或不存在;Z为氧,CH2或NH;n=l,2,3,4,5或6。优选的,R选自氢,R'选自氢或CH3;X为CONH;Y为不存在;Z为NH;n=l,2,3或4。本发明的另外一个目的是提供一种制备上述新霉胺-咔啉羧酸缀合物通式I或通式II的方法;一种制备上述通式I化合物的方法,包括以下步骤(1)将新霉胺分别进行叠氮反应和苯甲酰化反应得到0-[3\4'-二苯甲酰基-2',6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-l,3-二叠氮基-l,3-二脱氨-2-脱氧^^D-链霉胺;将0-[3',4、-二苯甲酰基-2',6、-二叠氮基-2、,6'-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-l,3-二叠氮基-l,3-二脱氨-2-脱氧一D-链霉胺游离羟基先三氟甲磺酸酯化,接着将三氟甲磺酸酯与相应的二胺氨解反应分别得到0-[3',4、-二苯甲酰基-2、,6、-二叠氮基-2',6、-二脱氧-0-0-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5"(2-氨基乙氨基)-1,3-二叠氮基-l,3-二脱氨-2,5-二脱氧一D-链霉胺,0-[3、,4:二苯甲酰基-2',6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-(4-氨基丁氨基)-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺,0-[3、,4、-二苯甲酰基-2、,6:二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]_6-苯甲酰基-5a-(6-氨基己氨基)-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺,0-[3、,4、-二苯甲酰基-2',6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5"(8-氨基癸氨基)-1,3-二叠氮基-l,3-二脱氨-2,5-二脱氧一D-链霉胺化合物;(2)将步骤(1)所得到的化合物分别与3S-2-叔丁氧羰酰基-l,2,3,4-四氢-e-咔啉-3-羧酸进行縮合反应,得到的中间体在室温脱出Boc保护,分别得到0-[3',4'-二苯甲酰基-2',6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)〗-6-苯甲酰基-5a-[(2-((3S-2-叔丁氧羰酰基-1,2,3,小四氢-e-昨啉-3-基)-羰酰氨基)乙基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧—D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(4-((35-2-叔丁氧羰酰基-1,2,3,4-四氢-e-咔啉-3-基)-羰酰氨基)丁基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺;0-[3、,(-二苯甲酰基-2',6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(6-((35-2-叔丁氧羰酰基-1,2,3,4-四氢-P-咔啉-3-基)-羰酰氨基)己基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(8-((3S-2-叔丁氧羰酰基-1,2,3,4-四氢-e-咔啉-3-基)-羰酰氨基)癸基)氨基]隱1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺;(3)将步骤(2)所得到的化合物在甲醇钠的甲醇溶液中皂化,随后还原叠氮基,即得;一种制备上述通式II化合物的方法,包括以下步骤(1)将新霉胺分别进行叠氮反应和苯甲酰化反应得到0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-1,3-二叠氮基-l,3-二脱氨-2-脱氧一D-链霉胺;将0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-l,3-二叠氮基-l,3-二脱氨-2-脱氧一D-链霉胺游离羟基先三氟甲磺酸酯化,接着将三氟甲磺酸酯与相应的二胺氨解反应分别得到0-[3',4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-(1-0-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5。-(2-氨基乙氨基)-1,3-二叠氮基-l,3-二脱氨-2,5-二脱氧一D-链霉胺,0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5ci-(4-氨基丁氨基)-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺,0-[3',4'-二苯甲酰基-2',6'-二叠氮基-2',6'-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-(6-氨基己氨基)-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺,0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2\6'-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5"(8-氨基癸氨基)-1,3-二叠氮基l,3-二脱氨-2,5-二脱氧一D-链霉胺;(2)将步骤(i)所制备得到的化合物分别与e-昨啉-3-羧酸和l-甲基-e-咔啉-3-羧酸进行縮合,分别得到以下化合物0-[3、,4、-二苯甲酰基-2、,6'-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(2-((卜咔啉-3-基)-羰酰氨基)乙基)氨基]-l,3-二叠氮基-l,3-二脱氨-2,5-二脱氧一D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(4-((0-咔啉陽3-基)-羰酰氨基)丁基)氨基]-l,3-二叠氮基-l,3-二脱氨-2,5-二脱氧一D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2',6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(6-((e-咔啉-3-基)-羰酰氨基)己基)氨基]-l,3-二叠氮基-l,3-二脱氨-2,5-二脱氧^D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6'-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(8-((P-咔啉-3-基)-羰酰氨基)癸基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧—D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基.(1—4)]-6-苯甲酰基-5a-[(2-((l-甲基-{3-咔啉-3-基)-羰酰氮基)乙基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧—D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-0!-0-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(4-((1-甲基-e-昨啉-3-基)-羰酰氨基)丁基)氨基]隱1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、國二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(6-((1-甲基-e-昨啉-3-基)-羰酰氨基)己基)氨基]-1,3-二叠氮基-1,3-脱氨-2,5-二脱氧—D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基.(1—4)]-6-苯甲酰基-5a-[(8-((1-甲基-e.咔啉-3-基)-羰酰氨基)癸基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧—D-链霉胺;其中,该縮合反应以HOBT/TBTU做縮合剂,在DIPEA存在下依次进行缩合反应;(3)将步骤(2)所制备得到的化合物在甲醇钠的甲醇溶液中皂化,随后还原叠氮基,即得。本发明采用咔啉羧酸及其衍生物作为平面芳稠环结构,通过连接臂与新霉胺缀合,所获得的缀合物具有协同作用,可增强与细菌16SrRNA和HIVTARRNA的结合能力。咔啉环通过嵌插作用,新霉胺通过静电、氢键形式,两者作用相辅相成,活性筛选结果显示,本发明化合物的活性优于一些现有药物。体外抑菌试验结果表明,本发明化合物具有优良的抑菌活性,说明本发明化合物可制备成抗菌药物,可作为临床潜在的抗菌制剂;本发明采用表面等离子共振技术(surfaceplasmonresonance,SPR)测定了本发明化合物与HIVTARRNA的结合能力,计算出本发明化合物与RNA的解离常数(dissociationconstants,《d,pM),试验结果表明,本发明化合物具有抗艾滋病病毒活性的潜力,可作为临床潜在的抗艾滋病制剂。本发明的又一目的是提供一种含有本发明上述通式化合物的药用组合物,该药物组合物由治疗上有效剂量的本发明缀合物与药学上可接受的赋型剂或者辅加剂组成;即将有效量的本发明缀合物与药学上可接受的载体或稀释剂配合后,按本领域常规的制剂方法将其制备成任意一种适宜的药物组合物。通常该组合物适合于口服给药和注射给药,也适合其他的给药方法。该组合物可以是片剂、胶囊剂、粉剂、颗粒剂、锭剂、栓剂,或口服液等液体制剂形式。根据不同的给药方法,本发明药物组合物可以含有0.1%-99%重量,优选10-60%重量的本发明化合物。图l本发明化合物(9a-9d,10a-10d,lla-lld)的合成路线图;反应条件(a).T肌,CuS04,Et3N,CH3CN;(b).C6H5C0C1,pyridine,54%;(c).Tf20,pyridine,CH2C12;(d).NH2(CH2)NH2,n=2,4,6,8,CH3CN,收率40%得到3a,48%得到3b,47%得到3c,47%得到3d;(f).TBTU,H0BT,DIPEA,DMF;65%得到6a,70%得到6b,50%得到6c,74%得到6d,67。/G得到7a,60。/。得到7b,66。/。得到7c,79%得到7d;(g).HC1/CH3C00C2H5,37%得到8a,44W得到8b,75X得到8c,83%得到8d.;(h).CH30H/CH30Na;(i).H2S,Py/Et3N/H20,70%。具体实施例方式下面结合具体实施例来进一步描述本发明,本发明的优点和特点将会随着描述而更为清楚。但这些实施例仅是范例性的,并不对本发明的范围构成任何限制。本领域技术人员应该理解的是,在不偏离本发明的精神和范围下可以对本发明技术方案的细节和形式进行修改或替换,但这些修改和替换均落入本发明的保护范围内。实施例l化合物2的制备(0-[3、,4、-二苯甲酰基-2、,6'-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-l,3-二叠氮基-l,3-二脱氨-2-脱氧-D-链霉胺)。C下,向悬浮液中缓慢滴加4.3mLTf20,冰浴下继续搅拌反应2小时。过滤除去不溶物,冰浴下将滤液滴入溶有2.0g新霉胺,29mgCuS04和3.6mLEt3N的6mL水溶液中,反应液搅拌自然升至室温并继续反应6小时。停止反应,加入乙酸乙酯,以饱和食盐水洗三次,酯层以无水硫酸钠干燥,过滤,蒸干。残留褐色固体溶于5mL吡啶,Or下,滴力口3.0mL(3.63g,25.9mmol)苯甲酰氯,室温下反应5小时。蒸干溶剂,残留物经硅胶助层析纯化(洗脱剂石油醚/乙酸乙酯4:1)得到白色粉末2.5g,收率54%。&NMR(500MHz,CDC13):<58.09-8.11(m,2H),7.91-7.93(m,4H),7.58-7.61(m,1H),7.46-7.54(m,4H),7.35-7.39(m,4H),5.97(dd,J=9.5,10.5Hz,1H),5.58(d,/=4.0Hz,1H,Hl,),5.48(t,/=9.8Hz,1H),5.22(t,/=9.8Hz,1H),4.53-4.57(m,1H),3.92(td,■/=3.5,9.3Hz,1H),3.80(dd,J=3.5,10.5Hz,1H),3.71-3.76(m,1H),3.61-3.65(m,2H),3.53-3.58(m:1H),3.49(dd,J二3.0,13.5Hz,1H),3.42(dd,J"=5.5,9.0Hz,1H),2.51(dt,=13,0Hz,^=力=5.0Hz,1H,H2eq),1.75(q,/=13.0Hz,1H,H2ax).13CNMR(75MHz,CDC13):<5166.15,165.67,165.28,133.61,133.54,129.99,129.87,129.80,128.98,128.63,128.52,128.46,128,44,98.94,83.23,75.63,74.83,71.01,69.99,69.67,62.12,58.49,58.26,50.96,32.08.HRMS(ESI)forC33H3QN1209理论值761.2151[(M+Na)+];实验值761.2166.实施例2化合物3a-d的制备1、化合物3a的制备(0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a一(2-氨基乙氨基)-1,3-二叠氮基-l,3-二脱氨-2,5-二脱氧-D-链霉胺)化合物2(1.0g,1.35mmol)溶于5mL二氯甲垸中,加入lmL吡啶,冰浴下,1.5小时内向溶液中缓慢加入0.5mLTf20(0.84g,3.00mmol),加毕,继续反应2.5小时。停止反应,加入乙酸乙酯,以饱和食盐水洗三次,有机层以无水硫酸钠干燥,过滤,减压蒸干,残留物经硅胶助层析纯化(洗脱剂石油醚/乙酸乙酯8:1)得到淡黄色粉末。将所得的淡黄色粉末溶于6mL乙睛中,加入4.05mmolNH2(CH2)2NH2,室温搅拌过夜。停止反应,加入二氯甲烷,以饱和食盐水洗三次,有机层以无水硫酸钠干燥,过滤,减压蒸干,残留物经硅胶柱层析纯化得到产品化合物3a。化合物3a(白色粉末,收率40%,洗脱剂氯仿/甲醇30:1):^NMR(300MHz,CDC13)(58.16-8.19(m,2H),7.87-7.92(m,4H),7.43-7.52(m,5H),7.26-7.36(m,4H),5.99(t,9.9Hz,1H),5.52(t,J=9.8Hz,1H),5.21(d,J=3.6Hz,1H,Hl,),5.02(dd,J=2.1,10.5Hz,IH),4.68-4.78(m,IH),4.48-4.54(m,IH),4.27-4.36(m,IH),3.86(dd,■/=3.3,10.8Hz,2H),3.72(t,IH,H5),3.54(dd,J"=2.7,13.5Hz,IH),3.40(dd,>/=5.1,13.5Hz,IH),2.99-3.11(m,3H),2.82-2.96(m,IH),2.51(dt,/=4.8,12.9Hz,IH,H2eq),1.47(q,J=12.3Hz,IH,H2ax).13CNMR(75MHz,CDC13)5165.53,165.38,165.24,133.60,133.41,130.05,129.86,129.75,128.79,128.54,128.44,128.35,94.46,78.10,70.93,70.08,69.33,61.75,57.55,56.36,56.07,50.73,41.12,32.62.HRMS(ESI)forC35H36N1408理论值781.2913[(M+H)+];实验值781.2899.2、化合物3b的制备(0-[3、,4、-二苯甲酰基-2',6、-二叠氮基-2、,6—-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-(4-氨基丁氨基)1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧-D-链霉胺)与化合物3a的制备不同之处在于将所得的淡黄色粉末溶于6mL乙睛中,加入4.05mmolNH2(CH2)4NH2;其余均与化合物3a的制备完全相同;(白色粉末,收率48%,洗脱剂二氯甲烷/甲醇25:1):&NMR(500MHz,CDC13)<58.08-8.09(m,2H),7.91-7.94(m,4H),7.60-7.62(m,1H),7.48-7.53(m,4H),7.35-7.39(m,4H),5.97(dd,9.5,10.5Hz,1H),5.54(t,</=9.8Hz,1H),5.20(d,J=4.0Hz,1H,Hl,),4.94(dd,J=2.5,10.5Hz,1H),4.49-4.52(m,1H),4.37-4.42(m,1H),4.08-4.14(m,1H),3.81-3.87(m,2H),3.67(t,/=2.8Hz,1H,H5),3.54(dd,《/=2.5,10.5Hz,1H),3.41(dd,/=5.5,14.0Hz,1H),2.84-2.89(m,1H),2.69-2.71(m,2H),2.44-2.55(m,4H),1.40-1.56(m,5H).13CNMR(75MHz,CDC13)3165.65,165.49,165.23,133.63,133.55,129.87,129.78,129.00,128.65,128.46,128.42,94.20,77.72,77.25,70.91,70.07,69.27,61.76,57.68,56.58,56.10,50.74,50.52,41.58,32.55,30.53,27.90.HRMS(ESI)forC37H40N14O8理论值809.3226[(M+H)+];实验值809.3233.3、化合物3c的制备(0-[3',4、-二苯甲酰基-2、,6:二叠氮基-2、,6'-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-(6-氨基己氨基)1,3-二叠氮基-1,3-二脱11氨-2,5-二脱氧-D-链霉胺)与化合物3a的制备不同之处在于将所得的淡黄色粉末溶于6mL乙睛中,加入4.05mmolNH2(CH2)6NH2;其余均与化合物3a的制备完全相同;化合物3c(白色粉末,收率47%,洗脱剂二氯甲烷/甲醇15:1)。&NMR(500MHz,CDC13)<58.05-8.07(m,2H),7.90-7.92(m,4H),7.56-7.60(m,1H),7.44-7.52(m,4H),7.32-7.38(m,4H),5.98(dd,/=9.0,10.0Hz,1H),5.52(t,9.8Hz,1H),5.19(d,J=4.0Hz,1H,Hl,),4.91(dd,J=2.5,10.5Hz,1H),4.45-4.49(m,1H),4.33-4.38(m,1H),4.05-4.10(m,1H),3.80-3.85(m,2H),3.68(t,■/=3.0Hz,1H,H5),3.53(dd,/=2.5,10.5Hz,1H),3.41(dd,J=5.0,10.5Hz,1H),2.85(t,J=7.8Hz,2H),2.74-2.79(m,IH),2.48-2.53(m,IH),2.43(dt,J=5.0,13.5Hz,1H),1.53-1.57(m,2H),1.36-1.45(m,3H),1.19-1.26(m,7H).13CNMR(75MHz,CDC13)3165.68,165.55,165.26,133.72,133.68,133.57,129.89,129.79,129.09,128.66,128.60,128.51,128.46,93.93,77.42,77.32,70.91,69.98,69.44,61.53,57.85,56.67,55.66,50.81,50.37,40.41,32.59,30.16,27.25,26.50,25.90.H腹S(ESI)forC39H44Ni408理论值837.3539[(M+H)+];实验值837.3545.4、化合物3d的制备(0-[3、,4、-二苯甲酰基-2',6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-50-(8-氨基癸氨基)1,3-二叠氮基-l,3-二脱氨-2,5-二脱氧-D-链霉胺)与化合物3a的制备不同之处在于将所得的淡黄色粉末溶于6mL乙睛中,加入4.05mmolNH2(CH2)8NH2;其余均与化合物3a的制备完全相同;化合物3d(白色粉末,收率47%,洗脱剂氯仿/甲醇15:1):^NMR(500MHz,CDC13)(58.06-8.08(m,2H),7.90-7.92(m,4H),7.53-7.60(m,1H),7.44-7.52(m,4H),7.33-7.38(m,4H),6.9-7.20(br,2H),6.00(t,/=9.5Hz,1H),5.52(t,J=9.5Hz,1H),5.20(d,J=3.5Hz,1H,Hl,),4.93(dd,/=2.5,10.5Hz,1H,H6),4.45-4.49(m,1H),4.38(dt,/=5.0,12.0Hz,1H),4.09(dt,/=5.0,11.5Hz,1H),3.82-3.86(m,2H),3.68-3.70(t,1H,H5),3.53(dd,/=2.5,13.5Hz,IH),3.41(dd,/=5.0,8.5Hz,1H),2.91(t,/=7.8Hz,2H),2.79-2.81(m,1H),2.52(br,1H),2.43(dt,■/=5.0,13.5Hz,1H),1.55-1.61(m,2H),1.40-1.47(m,3H),0.92-1.30(m,9H).13CNMR(75MHz,CDC13)(5165.53,165.28,133.66,133.53,129.90,129.83,129.78,129.10,128.62,128.49,128.44,93.94,77.47,70.91,70.01,69.47,61.58,57.86,56.67,55.78,50.80,50.64,40.38,32.61,30.37,29.02,28.70,27.20,27.00,26.00.HRMS(ESI)forC41H48N1408理论值865.3852[(M+H)+];实验值865.3902.实施例3化合物6a-d,7a-d,和8a-d的合成化合物6a的制备(0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(2-(e-昨啉-3-基)-羰酰氨基)乙基)氨基]-l,3-二叠氮基-1,3-二脱氨-2,5-二脱氧-D-链霉胺)将0.15mmo1咔啉羧酸衍生物(4a:P-咔啉-3-羧酸),0.14mmol3a,0.18mmolHBTU((9-(1//-苯并三唑-1-基)^凡忖'^-四甲基异脲六氟化磷),和0.18腿olHOBT(l-羟基-苯骈三唑)溶于5mL二甲基甲酰胺溶液中,搅拌下加入0.3mLDIPEA(二异丙基乙基胺),TLC检测至反应结束。蒸干溶剂,残留物溶于二氯甲烷中,减压浓縮,残留物残留物经硅胶助层析(洗脱剂二氯甲垸/氯仿)纯化得到产品6a。化合物6a(收率65%):^NMR(300MHz,CDC13)38.94-8.96(m,2H),8.81(m,1H),8.65-8.68(m,1H),8.18(d,/=7.8Hz,1H),8.02(d,/=7.2Hz,2H),7.86-7.97(m,4H),7.44-7.60(m,5H),7.21-7.42(m,8H),6.01(t,■/=9.9Hz,1H),5.50(t,J=9.8Hz,1H),5.24(d,/=3.6Hz,1H,Hl,),4.97(dd,/=2.4,10.2Hz,IH),4.47-4.56(m,2H),4.04-4.09(m,IH),3,74-3.89(m,4H),3.37-3.55(m,3H),3.06-3.09(m,1H),2.89-2.94(m,1H),2.44(dt,/=5.0,12.9Hz,1H,H2叫),1.43(q,J=12.9Hz,1H,H2股).13CNMR(75MHz,CDC13)<5165.51,165.42,165.29,140.67,140.44,137.07,133.65,133.48,131.91,129.89,129.75,129.50,128.89,128.63,128.54,128.49,128.40,122.17,121.81,120.84,114.38,111,71,94.19,70.86,70.05,69.52,61.56,57.68,56.91,55.39,50.81,49.86,39.92,32.58.HRMS(ESI)forC47H42N1609理论值975.3393[(M+H)+];实验值975.3411.2.化合物6b的制备(0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6'-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5"[(4-(e-咔啉-3-基)-羰酰氨基)丁基)氨基]陽l,3-二叠氮基-1,3-二脱氨-2,5-二脱氧-D-链霉胺)与化合物6a的制备不同之处在于将0.15mmo1咔啉羧酸衍生物(4a:P-咔啉-3-羧酸),0.14mmol3b,0.18mmolHBTU苯并三唑-l-基)-N,N,N',N'-四甲基异脲六氟化磷),和0.18mmolHOBT(l-羟基-苯骈三唑)溶于5mL二甲基甲酰胺溶液中,搅拌下加入0.3mLDIPEA(二异丙基乙基胺),其余均与化合物6a的制备完全相同;化合物6b(收率70%):力NMR(500MHz,CDC13)(58.90(s,1H),8.81(s,1H),8.73(d,/=1.0Hz,1H),8.24(t,/=6.0Hz,1H),8.17(d,J=8.0Hz,1H),8.07-8.09(m,2H),7.90-7.93(m,4H),7.56-7.59(m,2H),7.45-7.53(m,5H),7.31-7.38(m,5H),6.03(dd,J=9.0,10.5Hz,1H),5.52(t,J=10.0Hz,1H),5.21(d,/=4.0Hz,1H,Hl,),4.94(dd,■/=2.5,10.5Hz,IH),4.46-4.49(m,1H),4.36-4,42(m,1H),4.07-4.13(m,IH),3.81-3.86(m,2H),3.69(t,/=2.8Hz,1H,H5),3.48-3.55(m,3H),3.41(dd,</=5.0,14.0Hz,1H),2.85-2.90(m,1H),2.61-2.66(m,1H),2.43(dt,■/=13.0,5.0Hz,1H,H2叫),1.56-1.72(m,5H),1.43(q,/=13.0Hz,lH,H2狀).13CNMR(75MHz,CDC13)<5165.54,165.46,165.28,140.68,140.51,137.06,133.67,133.64,133.46,131.67,129.90,129.80,129.54,129.04,128.86,128.70,128.49,128.42,122.17,121.83,120.81,114.35,111.67,93.92,77.48,77.25,70.94,70.00,69.49,61.57,57.89,56.66,55.93,50.81,50.33,39.16,32.67,27.95,27.58.H謹S(ESI)forC49H46N1609理论值1003.3706[(M+H)+];实验值1003.3708.3.化合物6c的制备(0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(6-(e-咔啉-3-基)-羰酰氨基)己基)氨基]-l,3-二叠氮基-1,3-二脱氨-2,5-二脱氧-D-链霉胺)与化合物6a的制备不同之处在于将0.15mmo1咔啉羧酸衍生物(4a:e-昨啉-3-羧酸),0.14mmol3c,0.18mmolHBTU(0-(1//-苯并三唑-1-基)^^^,^-四甲基异脲六氟化磷),14和0.18mmolHOBT(l-羟基-苯骈三唑)溶于5mL二甲基甲酰胺溶液中,搅拌下加入0.3mLDIPEA(二异丙基乙基胺),其余均与化合物6a的制备完全相同;化合物6c(收率50%):NMR(500MHz,CDC13)39.02((!,/=3.0Hz,1H),8.92(s,1H),8.76(d,/=1.0Hz,1H),8.13-8.18(m,2H),8.07-8.09(m,2H),7.91-7.92(m,4H),7.46-7.60(m,7H),7.31-7.38(m,5H),6.01(dd,J=9.5,10.5Hz,IH),5.51(t,J=10.0Hz,IH),5.18(d,J"=4.0Hz,IH,Hl,),4.93(dd,J=2.0,10,0Hz,IH),4.45-4.48(m,IH),4.35-4.41(m,IH),4.05-4.10(m,IH),3.79-3.84(m,2H),3.67(t,J=3.0Hz,1H,H5),3.52(dd,/=2.5,13.5Hz,IH),3.46(q,/=7.0Hz,2H),3.40(dd,J=5.5,14.0Hz,IH),2.79-2.84(m,IH),2.50-2.55(m,IH),2.43(dt,J=13.5,5.0Hz,IH,H2eq),1.73(br,IH),1.56-1.61(m,2H),1.38-1.50(m,3H),1.26-1.32(m,4H),13CNMR(75MHz,CDC13)3165.53,165.42,165.30,140.74,140.50,137.09,133.68,133.64,133.47,131.67,129.89,129.78,129.56,129.09,128.87,128.65,128.48,128.41,122.15,121.83,120.80,114.36,111.73,93.91,77.48,77.26,70.91,69.97,69.50,61.54,57.86,56.66,55.80,50.81,50.61,39.39,32.66,30.41,29.62,26.93,26.80.HRMS(ESI)forC5iH5qN1609理论值1031.4019[(M+H)+];实验值1031.4028.4.化合物6d的制备(0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D.吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(8-(e-昨啉-3-基)-羰酰氨基)癸基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧-D-链霉胺):与化合物6a的制备不同之处在于将0.15mmo1咔啉羧酸衍生物(4a:P-咔啉-3-羧酸),0.14mmol3d,0.18mmolHBTU(O-(l乐苯并三唑-l-基)-N,N,N',N'-四甲基异脲六氟化磷),和0.18mmolHOBT(l-羟基-苯骈三唑)溶于5mL二甲基甲酰胺溶液中,搅拌下加入0.3mLDIPEA(二异丙基乙基胺),其余均与化合物6a的制备完全相同;化合物6d(收率74%):&NMR(500MHz,CDC13)<59.06(s,1H),8.93(s,1H),8.77(d,J=1.0Hz,1H),8.14-8.18(m,2H),8.07-8.09(m,2H),7.91-7.93(m,4H),7.45-7.60(m,7H),7.31-7.38(m,5H),6.01(dd,J=9.5,10.5Hz,1H),5.51(t,J-10.0Hz,1H),5.17(d,3.5Hz,1H,Hl,),4.93(dd,/=2.5,10.5Hz,IH),4.45-4.48(m,IH),4.36-4.41(m,IH),4.05-4.10(m,IH),3.79-3.84(m,2H),3.67(t,/=3.0Hz,1H,H5),3.47-3.54(m,3H),3.40(dd,/=5.0,13.5Hz,IH),2.79-2.84(m,IH),2,41-2.51(m,2H),1.66(br,IH),1.57-1.62(m,2H),1.16-1.47(m,11H).13CNMR(75MHz,CDC13)<5165.57,165.53,165.44,165.31,140.75,140.51,137.09,133.64,133.49,131.66,129.89,129.81,129.77,129.57,129.09,128.86,128.62,128.48,128.42,122.15,121.83,120.78,114.37,111.73,93.89,77.49,77.31,70.92,69.98,69.50,61.56,57.86,56.67,55.82,50.80,50.70,39.48,32.65,30.47,29.63,29.21,29.17,27.12,26.89.HRMS(ESI)forC53H54N1609理论值1059.4332[(M+H)+];实验值1059.4344.5.化合物7a的制备(0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-。-0-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(2-(1-甲基-e-咔啉-3-基)-羰酰氨基)乙基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧-D-链霉胺):与化合物6a的制备不同之处在于将0.15mmo1咔啉羧酸衍生物(4b:1—甲基-0-咔啉-3-羧酸),0.14mmol3a,0.18mmolHBTU(0-(1//-苯并三唑-1-基)^,:^^[',:^-四甲基异脲六氟化磷),和0.18mmolHOBT(l-羟基-苯骈三唑)溶于5mL二甲基甲酰胺溶液中,搅拌下加入0.3mLDIPEA(二异丙基乙基胺),其余均与化合物6a的制备完全相同;化合物7a(收率67%):NMR(300MHz,CDC13)39.23(br,1H),8.82(s,1H),8.70-8.74(m,1H),8.11(d,J=7.8Hz,1H),8.01(d,J=7.2Hz,2H),7.85-8.00(m,4H),7.22-7.53(m,12H),6.02(t,J=9.9Hz,IH),5.52(t,/=9.9Hz,IH),5.25(d,J=3.6Hz,IH,Hl,),5.00(dd,J=2.7,10.2Hz,IH),4.47-4.55(m,2H),4.08-4.15(m,IH),3.76-3.91(m,4H),3.38-3.56(m,3H),2.99-3.08(m,2H),2.82(s,3H,CH3),2.44(dt,/=5.1,13.2Hz,IH,H2eq),1.85(br,IH),1.46(q,/=12.6Hz,IH,H2狀).13CNMR(75MHz,CDC13)5165.86,165.37,165.22,140.50,139.27,136.05,133.55,133.37,129.76,129.66,128.56,128.44,128.27,122.15,121.96,121.88,120.47,112.58,111,82,94.02,77.42,76.57,70.72,69.93,69.40,61.40,57.60,56.46,55.30,50.69,49.80,39.77,32.37,20.31.HRMS(ESI)forC48H44N1609理论值989.3550[(M+H)+];实验值989.3582.6.化合物7b的制备(0-[3、,4、-二苯甲酰基-2、,6'-二叠氮基-2',6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(4-(1-甲基-e-咔啉-3-基)-羰酰氨基)丁基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧-D-链霉胺)与化合物6a的制备不同之处在于将0.15mmo1咔啉羧酸衍生物(4b:1-甲基-0-咔啉-3-羧酸),0.14腿0131>,0.18mmolHBTU(CKlif-苯并三唑-l-基)-N,N,N',N'-四甲基异脲六氟化磷),和0.18mmolHOBT(l-羟基-苯骈三唑)溶于5mL二甲基甲酰胺溶液中,搅拌下加入0.3mLDIPEA(二异丙基乙基胺),其余均与化合物6a的制备完全相同;化合物7b(收率60%):&NMR(500MHz,CDC13)58.77(s,1H),8.41(s,1H),8.23(t,J=6.3Hz,1H),8.15(d,/=8.0Hz,1H),8.08-8.10(m,2H),7.89-7.92(m,4H),7.46-7.60(m,7H),7.31-7.38(m,5H),6.00(dd,9.5,10.5Hz,1H),5.50(t,/=10.0Hz,1H),5.21(d,J=4.0Hz,1H,HI'),4.94(dd,/=2.5,10.5Hz,1H),4.45-4.49(m,1H),4.37-4.43(m,1H),4.06-4.12(m,161H),3.80-3.86(m,2H),3.69(t,/=3.0Hz,1H,H5),3.47-3.54(m,3H),3.40(dd,/=5.0,14.0Hz,1H),2.87-2.92(m,1H),2.76(s,3H,CH3),2.61-2.65(m,1H),2.44(dt,J=13.0,5.0Hz,1H,H2eq),1.56-1.73(m,5H),1.43(q,J=12.5Hz,lH,H2狀).13C画R(75MHz,CDC13)3165.56,165.53,165.44,165.29,140.25,140.03,139.97,135.78,133.67,133.63,133.43,129.92,129.83,129.80,129.07,128.83,128.70,128.56,128.49,128.39,122.48,122,20,120.85,112,68,111.64,94.01,77.58,77.26,70.88,70.02,69.52,61.58,57.89,56.64,55.95,50.84,50.37,39.28,32.69,28.10,27.75,20.19.HRMS(ESI)forC50H48N16O9理论值1017.3863[(M+H)+];实验值1017.3850.7.化合物7c的制备(0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D國吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(6-(1-甲基-e-咔啉-3-基)-羰酰氨基)己基)氨基]-1,3-二叠氮基-1,3-脱氨-2,5-二脱氧-D-链霉胺):与化合物6a的制备不同之处在于将0.15mmo1昨啉羧酸衍生物(4b:1-甲基-0-咔啉-3-羧酸),0.14譲ol3c,0.18腿olHBTU((9-(1//-苯并三唑-1-基)^^,"-四甲基异脲六氟化磷),禾口0.18mmolHOBT(l-羟基-苯骈三唑)溶于5mL二甲基甲酰胺溶液中,搅拌下加入0.3mLDIPEA(二异丙基乙基胺),其余均与化合物6a的制备完全相同;化合物7c(收率66%):!HNMR(500MHz,CDC13)<59.00-9.01(m,1H),8.78(s,1H),8.23(t,/=5.5Hz,1H),8.07-8.11(m,3H),7.91-7.92(m,4H),7.45-7.59(m,7H),7.25-7.37(m,5H),6.00(dd,J=9.5,10.0Hz,1H),5.51(t,/=10.0Hz,1H),5.17((!,/=3.5Hz,1H,Hl,),4.93(dd,J=2.5,10.0Hz,IH),4.45-4.49(m,IH),4.36-4.41(m,IH),4.05-4.10(m,IH),3.79-3.84(m,2H),3.68(t,J=2.8Hz,IH,H5),3.38-3.54(m,4H),2.79-2.84(m,4H),2.50-2.55(m,IH),2.43(dt,/=13.5,5.0Hz,IH,H2eq),1.58-1.61(m,3H),1.39-1.47(m,3H),1.26-1.31(m,4H).13CNMR(75MHz,CDC13)<5165.58,165.54,165.47,165.29,140.38,140.10,139.84,135.88,133.66,133.61,133.44,129.88,129.77,129.07,128.74,128.63,128.47,128.38,122.38,122.09,120.70,112.61,111.72,93.93,77.51,77.25,70.87,69.97,69.49,61.54,57.85,56.64,55.79,50.80,50.63,39.38,32.63,30.46,29.76,27.00,26.87,20.24.HRMS(ESI)forC52H52N1609理论值1045.4176[(M+H)+];实验值1045.4160.8.化合物7d的制备(0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D匿吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(8-(1-甲基-e-咔啉-3-基)-羰酰氨基)癸基)氨基]隱1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧-D-链霉胺):与化合物6a的制备不同之处在于将0.15mmo1咔啉羧酸衍生物(4b:l-甲基-g-咔啉-3-羧酸),0.14腿ol3d,0.18画olHBTU(O-(lH-苯并三唑-l-基)-N,N,N',N'-四甲基异脲六氟化磷),和0.18mmolHOBT(l-羟基-苯骈三唑)溶于5mL二甲基甲酰胺溶液中,搅拌下加入0.3mLDIPEA(二异丙基乙基胺),其余均与化合物6a的制备完全相同;化合物7d(收率79%):NMR(500MHz,CDC13)(59.01(s,1H),8.79(s,1H),8.24(t,/=6.0Hz,1H),8.07-8.11(m,3H),7.91-7.93(m,4H),7.45-7.59(m,7H),7.28-7.38(m,5H),6.01(dd,11.0,10.0Hz,1H),5.51(t,/=10.0Hz,1H),5.18(d,/=4.0Hz,1H,Hl,),4.93(dd,/=2.0,10.0Hz,1H),4,45-4.49(m,1H),4.36-4.41(m,1H),4.05-4.10(m,1H),3.80-3.84(m,2H),3.67(t,/=3.0Hz,1H,H5),3.47-3.54(m,3H),3.40(dd,J=5.5,13.5Hz,1H),2.79-2.84(m,4H),2.47-2.52(m,1H),2.43(dt,/=13.5,5.0Hz,1H,H2eq),1.58-1.64(m,2H),1.30-1.50(m,6H),1.17-1.28(m,6H).13CNMR(75MHz,CDC13)3165.61,165.54,165.49,165.29,140.42,140.13,139.84,135.92,133.62,133.47,129.88,129.79,129.76,129.09,128.74,128.61,128.46,128.40,122.38,122.08,120.67,112.61,111.74,93.91,77.51,77.26,70.91,69.98,69.49,61.57,57.85,56.66,55.83,50.80,50.67,39.48,32.64,30.51,29.76,29.28,29.24,27.19,26.96,20.29.HRMS(ESI)forC54H56N1609理论值1073.4489[(M+H)+];实验值1073.4484.9.化合物8a的制备(0-[3、,4'-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧巧-0-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(2-(35-2-叔丁氧羰酰基-1,2,3,4-四氢-P-咔啉-3-基)-羰酰氨基)乙基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧-D-链霉胺)与化合物6a的制备不同之处在于将0.15mmo1咔啉羧酸衍生物(5:3S-2-叔丁氧羰酰基-1,2,3,4-四氢-e-咔啉-3-羧酸),0.14腿ol3a,0.18mmolHBTU((9-(1//-苯并三唑陽1-基)-N,N,N',N'-四甲基异脲六氟化憐),和0.18mmolHOBT(l-羟基-苯骈三唑)溶于5mL二甲基甲酰胺溶液中,搅拌下加入0.3mLDIPEA(二异丙基乙基胺),其余均用与化合物6a的制备完全相同的方法可得到8a前体。之后将8a前体室温用lNHCl/乙酸乙酯处理脱Boc得到8a。化合物8a(收率37%):NMR(500MHz,CDC13)<58.05(dd,J=1.0,8.5Hz,2H),7.90-7.91(m,2H),7.83(dd,J"=1.0,8.5Hz,2H),7.40-7,55(m,6H),7.35-7.38(m,2H),7.26-7.30(m,4H),7.14-7.17(m,IH),7.09-7.12(m,IH),5.99(dd,J=9.5,10.5Hz,IH),5.51(t,/=10.0Hz,IH),5.21(d,/=3.5Hz,IH,HI'),4.96(dd,J=2.0,10.5Hz,IH),4.48-4.52(m,IH),4.33-4.38(m,IH),4.00-4.11(m,3H),3.81-3.85(m,2H),3.69(t,J=3.0Hz,1H,H5),3.63(q,/=4.7Hz,IH),3.51-3.58(m,2H),3.41(dd,5.5,13.5Hz,IH),3.26-3.32(m,IH),3.21(dd,■/=4.8,15.5Hz,1H),2.84-2.95(m,3H),2.41(dt,/=13.5,5.0Hz,1H,H2eq),1.82(br,2H),1.45(q,J=12.5Hz,lH,H2狀).13CNMR(75MHz,CDC13)<5172.63,165.65,165.47,165.26,135.96,133.80,133.67,133.54,129.88,129.78,129.75,128.84,128.75,128.50,128.42,127.37,121,84,119.63,118.13,110.73,94.41,77.77,70.92,70.12,69.42,61.81,57.62,57.13,56.73,55.83,50.83,49.54,42.72,39.84,32.46,24.32.HRMS(ESI)forC47H46N1609理论值979.3706[(M+H)+];实验值979.3856.10.化合物8b的制备(0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D.吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-["-(3S-2-叔丁氧羰酰基-1,2,3,小四氢-13陽咔啉-3-基)-羰酰氨基)丁基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧-D-链霉胺)与化合物6a的制备不同之处在于将0.15mmd咔啉羧酸衍生物(5:3S-2-叔丁氧羰酰基-1,2,3,4-四氢-0-咔啉-3-羧酸),0.14mmol3b,0.18mmolHBTU(O-(lif-苯并三唑-l-基)-N,N,N',N'-四甲基异脲六氟化磷),和0.18mmolHOBT(l-羟基-苯骈三唑)溶于5mL二甲基甲酰胺溶液中,搅拌下加入0.3mLDIPEA(二异丙基乙基胺),其余均用与化合物6a的制备完全相同的方法可得到8b前体。之后将8b前体室温用lNHCl/乙酸乙酯处理脱Boc得到8b。化合物8b(收率44%):&NMR(500MHz,CDC13)(58.07-8.09(m,2H),7.89-7.92(m,3H),7.77(br,1H),7.60(t,/=7.5Hz,1H),7.47-7.53(m,5H),7.26-7.38(m,5H),7.08-7.17(m,3H),5.99(t,/=10.0Hz,1H),5.51(t,/=10.0Hz,1H),5.18(d,/=3.5Hz,1H,Hl,),4.93(dd,/=2.5,10.5Hz,1H),4.46-4.49(m,1H),4.34-4.39(m,1H),4.04-4.10(m,1H),3.94-4.02(m,2H),3.82(dt,/=4.0,10.5Hz,2H),3.67(t,/=3.0Hz,1H,H5),3.51-3.55(m,2H),3.41(dd,/=5.0,13.5Hz,1H),3.20-3.31(m,3H),2.79-2.88(m,2H),2.57-2.60(m,lH),2.42(dt,J=13.5,5.0Hz,1H,H2eq),1.67(br,2H),1.39-1.60(m,5H).13CNMR(75MHz,CDC13)3172.40,165.56,165.26,135.95,133.71,133.66,133.53,132.11,129.89,129.81,129.79,129.04,128.72,128.62,128.49,128.46,127.36,121.81,119.60,118.12,110.68,108.66,94.12,77.69,70.94,70.01,69.43,61.62,57.83,57.25,56.65,55.92,50.81,50.15,42.82,38.93,32.62,27.87,27.41,24.49.HRMS(ESI)forC49H50N16O9理论值1007.4019[(M+H)+];实验值1007.4050.11.化合物8c的制备(0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6'-二脱氧-a-D國吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(6-(3S-2-叔丁氧羰酰基-1,2,3,小四氢-P-昨啉-3-基)-羰酰氨基)己基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧-D-链霉胺)与化合物6a的制备不同之处在于将0.15mmo1咔啉羧酸衍生物(5:3S-2-叔丁氧羰酰基-1,2,3,4-四氢-P-昨啉-3-羧酸),0.14薩ol3c,0.18mmolHBTU(<9-(1//-苯并三唑-1-基)-N,N,N',N'-四甲基异脲六氟化磷),和0.18mmolHOBT(l-羟基-苯骈三唑)溶于5mL二甲基甲酰胺溶液中,搅拌下加入0.3mLDIPEA(二异丙基乙基胺),其余均用与化合物6a的制备完全相同的方法可得到8c前体。之后将8c前体室温用lNHCl/乙酸乙酯处理脱Boc得至lj8c。化合物8c(收率75%):NMR(500MHz,CDC13)38.07-8.09(m,2H),7.90-7.92(m,5H),7.61(t,7.5Hz,1H),7.46-7.53(m,5H),7.29-7.38(m,5H),7.08-7.17(m,2H),7.02(br,1H),6.00(t,J=10.0Hz,1H),5.51(t,>/=10.0Hz,1H),5.18(d,J=3.5Hz,1H,H1,),4.93(dd,J=2.5,10.5Hz,1H),4.45-4.49(m,1H),4.35-4.40(m,1H),4.01-4.10(m,3H),3.79-3.85(m,2H),3.68(t,J=2.5Hz,1H,H5),3.53(dd,2.5,13.5Hz,2H),3.41(dd,J=5.0,13.5Hz,1H),3.20-3.24(m,3H),2.76-2.84(m,2H),2.49-2.53(m,lH),2.43(dt,/=13.5,5.0Hz,1H,H2eq),1.99(br,2H),1.39-1.46(m,5H),1.22-1.30(m,5H).13CNMR(75MHz,CDC13)3172.27,165.53,165.27,135.98,133.70,133.65,133.51,131.94,129.89,129.81,129.78,129,10,128.65,128.49,128.43,127.27,121.81,119.58,118.07,110.76,108.51,93.96,77.53,70.91,69,98,69.46,61,54,57.85,57.27,56.67,55.86,50.81,50.61,42.85,39.08,32.62,30.44,29.40,26.90,26.74,24.61.HRMS(ESI)forC51H54N1609理论值1035.4332[(M+H)+];实验值1035.4334.12.化合物8d的制备(0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-(1-0-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(8-(3S-2-叔丁氧羰酰基-1,2,3,小四氢-P-咔啉-3-基)-羰酰氨基)癸基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧-D-链霉胺)与化合物6a的制备不同之处在于将0.15mmo1咔啉羧酸衍生物(5:3S-2-叔丁氧羰酰基-1,2,3,4-四氢-0-昨啉-3-羧酸),0.14薩ol3d,0.18mmolHBTU(0-(1乐苯并三唑-1-基)-N,N,N',N'-四甲基异脲六氟化磷),和0.18mmolHOBT(l-羟基-苯骈三唑)溶于5mL二甲基甲酰胺溶液中,搅拌下加入0.3mLDIPEA(二异丙基乙基胺),其余均用与化合物6a的制备完全相同的方法可得到8d前体。之后将8d前体室温用lNHCl/乙酸乙酯处理脱Boc得到8d。化合物8d(收率83%):&NMR(500MHz,CDC13)38.06-8.09(m,2H),7.90-7.93(m,5H),7.59-7.63(m,1H),7.47-7.53(m,5H),7.33-7.38(m,4H),7.29(d,/=8.0Hz,1H),7.08-7.17(m,2H),7.01(t,J=5.5Hz,1H),6.00(dd,J=9.5,10.5Hz,1H),5.50(t,/=10.0Hz,1H),5.16((!,/=4.0Hz,1H,Hl,),4.93((!(!,/=2.5,10.0Hz,1H),4.45-4.48(m,1H),4.35-4.41(m,IH),3.98-4.10(m,3H),3.78-3.84(m,2H),3.67(t,/=2.5Hz,1H,H5),3.50-3.54(m,2H),3.41(dd,/=5.0,13.5Hz,1H),3.23-3.27(m,3H),2.74-2.85(m,2H),2.47-2.52(m,lH),2.43(dt:/=13.5,5.0Hz,1H,H2eq),1.64(br,2H),1.38-1.51(m,5H),1.20-1.23(m,8H).13CNMR(75MHz,CDC13)(5172.46,165.53,165.48,165.27,135.96,133.64,133.49,132.34,129.88,129.81,129.76,129.10,128.62,128.47,128.42,127.34,121.76,119.56,118.06,110.72,108.70,93.91,77.50,77.30,70,89,69.97,69.47,61.54,57.85,57.42,56.67,55.85,50.80,50.68,43.08,39.11,32.62,30.50,29.47,29.25,29.10,27.14,26.82,24.75.HRMS(ESI)forC53H58N1609理论值1063.4645[(M+H)+];实验值1063.4676.实施例4化合物9a-d,lOa-d和11a-d的制备1.化合物9a的制备(0-[2、,6、-二氨基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-5ct-[(2-(e-咔啉-3-基)-羰酰氨基)乙基)氨基]-2,5-二脱氧-D-链霉胺)化合物6a0.05mmol悬浮于15mL甲醇中,加入30%的甲醇钠的甲醇溶液1mL,室温搅拌过夜。减压蒸除溶剂得到白色固体。将所得白色固体溶于4ml吡啶、2ml三乙胺、lml水混合成的均相溶剂中,搅拌下通入H2S气体1个小时。待反应液颜色变为墨绿色后停止通气,继续室温反应过夜。减压除去溶剂,残渣用甲醇溶解后吸附在硅胶上,常压柱层析(氨水/甲醇)分离。得到的产物用水溶解后用稀盐酸调节pl^6左右,冷冻干燥,得到的产物性状为白色泡状固体9a。收率70%。化合物9a:丄HNMR(500MHz,D20)<58.68(d,/=0.5Hz,1H),8.50(s,1H),8.03(d,/=8.0Hz,1H),7.66械/=1.0,8.5Hz,1H),7.52(d,J=8.0Hz,1H),7.35械/=1.0,8.5Hz,1H):215.65(d,/=4.0Hz,1H,Hl,),4.25(dd,■/=3.5,11.0Hz,IH),4.11(dd,J=9.0,10.5Hz,IH),4.06(dd,J=3.0,10.5Hz,IH),3.95-4.00(m,2H),3.78-3.90(m,2H),3.72(t,J=6.5Hz,2H),3.51-3.60(m,3H),3.43-3.48(m,IH),3.35(dd,J=6.5,13.5Hz,IH),3.25-3.30(m,IH),2.57(dt,/=12.5,4.5Hz,IH,H2eq),1.88(q,J=12.5Hz,IH,H2収).13CNMR(75MHz,D20)3164.51,143.26,135.96,132.94,132.04,131.56,129.11,122.92,122.53,120.36,115.34,113.32,92.14,73.54,71.23,71.05,70.36,69.15,57.49,53,90,49.95,48.93,47.87,40.73,40.65,29.15.HRMS(ESI)forC26H38N806理论值559.2987[(M+H)+];实验值559.3047.2.化合物9b的制备(0-[2、,6、-二氨基-2',6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-5a-[(小(P-咔啉-3-基)-羰酰氨基)丁基)氨基]-2,5-二脱氧-D-链霉胺)与化合物9a制备方法不同之处在于起始物为6b,其余均与化合物9a制备方法相同。收率70%。化合物9b:!HNMR(500MHz,D20)38.39(s,IH),8.11(s,IH),7.86(d,J"=8.0Hz,IH),7.53(t,■/=7.5Hz,IH),7.37(d,J=8.5Hz,IH),7.23(t,■/=7.5Hz,IH),5.56(d,/=8.5Hz,IH,Hl,),4.24(dd,/=2.5,10.5Hz,IH),4.11((!(!,/=3.0,11.0Hz,IH),4.049.0,10.5Hz,IH),3.95-3.99(m,2H),3.71-3.78(m,2H),3.45-3.54(m,5H),3.28-3.32(m,2H),3.07-3.09(m,IH),2.55(dt,/=12.5,4.5Hz,IH,H2eq),1.86(q,/=12.5Hz,IH,H2狀),1.74(br,4H).13CNMR(75MHz,D20)3165.87,142.16,136.37,135.55,130.76,129.87,122.39,121.72,120.53,114.65,112.91,92.69,73.56,71.09,70.49,69.27,57.88,53.86,51.21,49.04,47.86,40.68,40.16,29,18,27.04,26.43.HRMS(ESI)forC28H42N806理论值587.3300[(M+H)+];实验值:587.3314.3.化合物9c的制备(0-[2',6、-二氨基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-5ci-[(6-(e-咔啉-3-基)-羰酰氨基)己基)氨基]-2,5-二脱氧-D-链霉胺)与化合物9a制备方法不同之处在于起始物为6c,其余均与化合物9a制备方法相同。收率70%。化合物9c:!HNMR(500MHz,D20)(58.29(s,1H),8.00(s,1H),7.80(d,/=7.5Hz,1H),7.48(t,/=7.5Hz,1H),7.31(d,J^8.0Hz,1H),7.19(t,/=7.5Hz,1H),5.52(d,/=3.5Hz,1H:HI,),4.22(d,/=10.5Hz,1H),4.12(dd,J=3.0,10.5Hz,IH),3.95-4.03(m,3H),3.67-3.76(m,2H),3.44-3.53(m,3H),3.38(t,/=7.0Hz,2H),3.25-3.31(m,2H),3.00-3.06(m,IH),2.54(dt,J"=13.0,4.5Hz,1H,H2eq),1.85(q,/=12.5Hz,IH,H2収),1.68(br,4H),1.45(br,4H).13CNMR(75MHz,D20)5166.01,141.91,136.37,135.94,130.94,130.47,129.52,122.25,121.52,120.54,114.45,112.78,93.05,73.69,71.15,70.48,70.09,69.41,58.06,53.89,51.75,49.08,47.82,40.70,40.46,29.24,28.55,26.74,26.70.HRMS(ESI)forC3。H46N806理论值615.3613[(M+H)+];实验值615.3624.4.化合物9d的制备(0-[2、,6'-二氨基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-5a-[(8-(e-咔啉-3-基)-羰酰氨基)癸基)氨基]-2,5-二脱氧-D-链霉胺)与化合物9a制备方法不同之处在于起始物为6d,其余均与化合物9a制备方法相同。收率70%。化合物9d:NMR(500MHz,D20)58.52(s,1H),8.27(s,1H),7.86(d,>/=8.0Hz,1H),7.57(t,J=7.5Hz,IH),7.38(d,J=8.5Hz,IH),7.26(t,/=7.5Hz,IH),5.67(d,■/=3.5Hz,IH,Hl,),4.51(d,10.0Hz,IH),4.33-4.37(m,2H),4.10(t,/=9.3Hz,IH),4.01-4.05(m,IH),3.80-3.85(m,2H),3.60-3.64(m,2H),3.34-3.52(m,6H),2.64(dt,J=13.0,4.5Hz,IH,H2eq),1.97-2,07(m,IH),1.80-1.86(br,2H),1.68-1.73(m,2H),1.42(br,8H).13CNMR(75MHz,D20)<5162,38,143.37,135.22,132.41,132.03,131.87,127.73,122.79,122.71,119.91,114.72,113.19,92.70,72.05,71.59,70.45,68.72,67.84,59.10,53.47,49.07,47.71,41.07,40.40,29.23,29.08,29,05,28.61,26.98,26,79,26.58,22.01.HRMS(ESI)forC32H50N8O6理论值643.3926[(M+H)+];实验值643.3914.5.化合物l0a的制备(O-[2、,6、-二氨基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-5a-[(2-(1-甲基-e-咔啉-3-基)-羰酰氨基)乙基)氨基]-2,5-二脱氧-D-链霉胺)与化合物9a制备方法不同之处在于起始物为7a,其余均与化合物9a制备方法相同。收率70%。化合物10a:!HNMR(500MHz,D20)<58.16(s,1H),7.90(d,/=7.5Hz,1H),7.56(t,/=7.5Hz,1H),7.40(d,J=8.5Hz,1H),7.26(t,/=7.5Hz,IH),5.61(d,■/=4.0Hz,IH,HI'),4.17(dd,/=3.5,11.0Hz,IH),4.09(dd,/=9.0,10.5Hz,IH),3.94-3.99(m,2H),3.75-3.87(m,3H),3.65(t,/=6.5Hz,2H),3.50-3.56(m,3H),3.31-3,37(m,2H),3.13-3.18(m,IH),2.62(s,3H,CH3),2.54(dt,/=12.5,4.8Hz,IH,H2eq),1.83(q,■/=12.5Hz,IH,H2狀).13CNMR(75MHz,D20)(5165.78,141.06,141.18,135.34,134.45,130.83,129.05,122.39,121.95,120.86,113.37,112.82,92.00,73.96,71.96,71.20,70.11,69.23,57.05,53.98,49.53,48.92,47.91,41.00,40.71,29.28,18.22.HRMS(ESI)forC27H40N8O6理论值573.3144[(M+H)+];实验值573.3121.6.化合物l0b的制备(O-[2、,6、匿二氨基國2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-5a-["-(1-甲基-e-昨啉-3-基)-羰酰氨基)丁基)氨基]-2,5-二脱氧-D-链霉胺):与化合物9a制备方法不同之处在于起始物为7b,其余均与化合物9a制备方法相同。收率70%。化合物10b:^NMR(500MHz,D20)37.94(s,1H),7.76(d,J=8.0Hz,1H),7.49(t,/=7.5Hz,IH),7.29(d,/=8.5Hz,IH),7.18(t,/=7.5Hz,IH),5.57(d,/=3.0Hz,IH,Hl,),4.24(d,J=10.0Hz,IH),4,12(dd,J=11.0,3.5Hz,IH),4.05(dd,J=10.0,9.0Hz,IH),3.95-3.99(m,2H),3.74-3.79(m,2H),3.40-3.53(m,5H),3.27-3.31(m,2H),3.03-3.15(m,IH),2.5612.5,4.5Hz,IH,H2eq),2,49(s,3H,CH3),1.88(q,J=12,5Hz,IH,H2狀),1.76(br,4H).13CNMR(75MHz,D20)3165.46,141.75,140.89,135.07,134.69,130.59,128.66,122.16,121.75,120.72,112.92,112.64,92.70,73.60,71.19,70.41,69.31,57.78,53.90,51.14,49.06,47.86,40.72,40.25,39.85,29.20,27.09,26.56,18.22.HRMS(ESI)forC29H44N806理论值601.3457[(M+H)+];实验值601.3454.7.化合物l0c的制备(O-[2',6'-二氨基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-5a-[(6-(1-甲基-e-咔啉-3-基)-羰酰氨基)己基)氨基]-2,5-二脱氧-D-链霉胺)与化合物9a制备方法不同之处在于起始物为7c,其余均与化合物9a制备方法相同。收率70%。化合物10c:丄HNMR(500MHz,D20)<58.51(s,1H),8.07(d,8.0Hz,1H),7.69(td,■/=1.0,8.0Hz,1H),7.54(d,/=8.0Hz,IH),7.38(t,J=7.5Hz,IH),5.69(d,/=3.5Hz,IH,HI'),4.54(d,/=9.5Hz,IH),4.37-4.39(m,2H),4.11(t,/=8.8Hz,IH),4.02-4.06(m,IH),3.81-3.86(m,2H),3.62-3.65(m,2H),3.49-3.53(m,4H),3.39-3.45(m,2H),2.89(s,3H,CH3),2.65(dt,J=13.0,4.5Hz,IH,H2eq),2.01(q,J=12.5Hz,IH,H2狀),1.86-1.91(m,2H),1.74-1.79(m,2H),1.51(br,4H).13CNMR(75MHz,D20)5162.15,143.61,140.34,134.89,132.60,131.46,131.16,123.03,120.50,113.93,113.28,92.75,71.80,71.63,70.38,68.69,67.78,59.21,53.44,49.06,47.70,40.98,40.36,29.06,28.52,26.69,26.59,26.29,16.33.HRMS(ESI)forC31H48N806理论值629.3770[(M+H)+];实验值629.3773.8.化合物10d的制备(O-[2、,6、-二氨基-2、,6、-二脱氧-a-0-吡喃葡萄糖基-(1—4)〗-5a-[(8-(1-甲基-e-咔啉-3-基)-羰酰氨基)癸基)氨基]-2,5-二脱氧-D-链霉胺)与化合物9a制备方法不同之处在于起始物为7d,其余均与化合物9a制备方法相同。收率70%。化合物10d:^NMR(500MHz,D20)<57.83(s,1H),7.66(d,/=7.5Hz,1H),7.44(t,/=7.5Hz,1H),7.21(d,/=8.0Hz,IH),7.14(t,J=7.5Hz,IH),5.55(d,J=3.5Hz,IH,Hl,),4.28(dd,3.0,10.5Hz,IH),4.17(dd,J=3.5,10.5Hz,IH),3.97-4.05(m,3H),3.67-3.77(m,2H),3.54(t,■/=9.5Hz,IH),3.47-3.51(m,2H),3.38-3.41(m,2H),3.27-3.35(m,2H),3.05-3.10(m,IH),2.56(dt,J=12.5,4.7Hz,IH,H2eq),2.44(s,3H,CH3),1.87(q,/=12.5Hz,IH,H2ax),1.68-1.70(m,4H),1.35-1.48(m,8H).13CNMR(75MHz,D20)3164.68,141.75,140.45,134.73,134.25,130.72,128.69,122.02,121.80,120.49,112.64,112.53,93.13,73.47,71.04,70.64,69.67,69.32,58.23,53.83,52.04,49.08,47.79,40.76,40.66,29.35,29.27,29.21,28.29,27.06,26.95,17.89.HRMS(ESI)forC33H52N806理论值657.4083[(M+H)+〗;实验值:657,4079.9.化合物lla的制备(0-[2、,6、-二氨基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-5a-[(2-((3S-1,2,3,4-四氢-e-咔啉-3-基)-羰酰氨基)乙基)氨基]-2,5-二脱氧-D-链霉胺)与化合物9a制备方法不同之处在于起始物为8a,其余均与化合物9a制备方法相同。收率70%。化合物lla:丄H雨R(500MHz,D20)(57.61((!,/=8.0Hz,1H),7,51(d,/=8.0Hz,1H),7.28(t,7.5Hz,1H),7.20(t,/=7.5Hz,1H),5.49(d,J=4.0Hz,1H,Hl,),4.55(d,J=16.0Hz,1H),4.48(d,J=16.0Hz,1H),4.29(dd,J=11.0,5.0Hz,1H),3.98-4.06(m,2H),3.88-3.96(m,2H),3.65-3.72(m,3H),3.38-3.53(m,6H),3.30(dd,J=13.5,7.5Hz,1H),3.07-3.16(m,2H),2.97-3.02(m,1H),2.4712.5,4.5Hz,1H,H2eq),1.74(q,/=12.5Hz,1H,H2収).13CNMR(75MHz,D20)(5170.79,137.24,126.86,126.25,123.38,120.58,118.81,112.54,105.54,92.70,74.64,72.07,71.45,70.05,69.89,57.38,56.61,54.29,49.69,49.11,47.79,41.33,40.86,29.90,23.80.HRMS(ESI)forC26H42N806理论值563.3300[(M+H)+];实验值563.3296.10.化合物llb的制备(0-[2、,6、-二氨基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-5a-[(小(GS-1,2,3,小四氢-e-咔啉-3-基)-羰酰氨基)丁基)氨基]-2,5-二脱氧-D-链霉胺)与化合物9a制备方法不同之处在于起始物为8b,其余均与化合物9a制备方法相同。收率70%。化合物lib:&NMR(500MHz,D20)57.61(d,/=8.0Hz,IH),7.51(dd,J=1.0,7.5Hz:IH),7.29(td,/=8.0,1.0Hz,IH),7.20(td,J=8.0,1.0Hz,IH),5.55(d,/=4.0Hz,IH,Hl,),4.60(dd,J=1.0,15.5Hz,1H),4.54((!,/=15.5Hz,1H),4.34(dd,J"=11.0,5.5Hz,1H),4.19(dd,/=10.8,3.3Hz,IH),4.02-4.08(m,2H),3.95-3.99(m,IH),3.88(t,J=8.5Hz,IH),3.70-3.79(m,2H),3.42-3.56(m,4H),3.29-3.40(m,3H),3.13-3.20(m,2H),2.91-2,95(m,IH),2.55(dt,J=12.5,4.5Hz,IH,H2eq),1.85(q,/=12.5Hz,IH,H2欲),1.61-1.67(m,4H).13CNMR(75MHz,D20)3170.01,137.31,126.25,126.16,123.46,120.63,118.85,112.56,105.50,92.69,73.88,71.22,70.90,70.31,69.38,57.70,56.64,53.95,50.90,49.05,47.88,41.32,40.75,39.94,29.31,26.83,26.66,23.79.HRMS(ESI)forC28H46N806理论值591.3613[(M+H)+];实验值591.3637.11.化合物llc的制备(0-[2、,6、-二氨基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-5a-[(6-(GS-1,2,3,4-四氢-e-咔啉-3-基)-羰酰氨基)己基)氨基]-2,5-二脱氧-D-链霉胺)与化合物9a制备方法不同之处在于起始物为8c,其余均与化合物9a制备方法相同。收率70%。化合物llc:NMR(500MHz,D20)S7.59(d,J=8.0Hz,1H),7.51(d,>/=8.0Hz,IH),7.28(t,J=7.5Hz,IH),7.19(t,7.5Hz,IH),5.36(d,/=3.5Hz,IH,Hl,),4.5016.0Hz,IH),4.42(d,J=15.5Hz,IH),4.15(dd,■/=11.0,5.0Hz,IH),3.93-4.01(m,3H),3.86(t,/=10.0Hz,IH),3.78(t,3.5Hz,IH,H5),3.64(dt,J=12.0,4.5Hz,IH),3.43-3.53(m,3H),3.32-3.38(m,2H),3.18-3.27(m,3H),3.05-3.15(m,2H),2.84-2.91(m,IH),2.42(dt,J=12.5,4.5Hz,IH,H2eq),1.67(q,/=12.5Hz,1H,H2股),1,45-1.57(m,4H),1.34(br,4H).13C雨R(75MHz,D20)(5170.56,137.21,127.21,126.32,123.30,120.55,118.77,112.51,105.63,94.51,75.36,71.77,71.19,71.04,69.82,58.29,56.64,54.63,51.81,49.37,47.73,41.33,41.06,40.11,30.67,29.00,28.91,26.67,26.50,23.91.HRMS(ESI)forC30H50N8O6理论值619.3926[(M+H)+];实验值619.3935.12.化合物lld的制备(0-[2、,6'-二氨基-2',6'-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-5a-[(8-((3S-1,2,3,4-四氢-e-咔啉-3-基)-羰酰氨基)癸基)氨基]-2,5-二脱氧-D-链霉胺)与化合物9a制备方法不同之处在于起始物为8d,其余均与化合物9a制备方法相同。收率70%。化合物lld:'HNMR(500MHz,D20)<57.60(d,J=8.0Hz,IH),7.51(d,J=7.5Hz,IH),7.28(t,/=7.5Hz,IH),7.20(t,/=7.5Hz,IH),5.46(d,/=3.5Hz,IH,Hl,),4.58(d,/=16.0Hz,1H),4.50((!,/=15.5Hz,1H),4.27(dd,/=10.0,5.0Hz,1H),4.12(dd,J=10.5,3.0Hz,1H),4.05(dd,/=10.5,3.0Hz,1H),3.94-3.98(m,2H),3.87(s,1H,H5),3.68(td,/=12.0,4.5Hz,IH),3.61(td,J=12.0,4.0Hz,1H),3.47-3.52(m,2H),3.12-3.42(m,7H),2.87-2.94(m,1H),2.49(dt,/=12.5,4.5Hz,1H,H2eq),1.78(q,J=12.5Hz,1H,H2ax),1.54-1.58(m,4H),1.28(br,8H).13CNMR(75MHz,D20)3169.81,137.26,126.34,126.20,123.40,120.59,118.82,112.53,105.45,93.51,74.37,71.45,70.56,70.09,69.98,57.95,56.46,54.16,51.62,49.16,47.82,41.19,40.88,40.25,29.73,29.17,28.97,28.92,27.00,26.60,23.63.HRMS(ESI)forC32H54N806理论值669.4058[(M+Na)+];实验值669.4062.试验例l本发明化合物的RNA结合活性试验1、供试化合物本发明实施例所制备的化合物,分别编号为9a,9b,9c,9d,10a,1Ob,10c,10d,11a,11b,11c,11d;2、试验方法及结果(1)采用表面等离子共振技术(surfaceplasmonresonance,SPR)分别测定了供试化合物与£coh16SrRNA和人18Sr脂ARNA的结合能力,计算出供试化合物与脂A的解离常数(dissociationconstants,,pM),试验结果如表l所示。从试验结果可见,供试化合物与16SrRNA和18Sr脂A的结合强于新霉胺,提示芳香性咔啉环在缀合物与RNA的结合中起到重要的作用。芳香环上甲基取代,活性稍下降。而当以四氢咔啉代替平面的咔啉环时,化合物lla,llb,llc,lld几乎与RNA无结合活性。连接链长短也对缀合物的活性有影响。当链长为2个或8个碳原子时(化合物9a和9d),活性最强,这可能是由于插入不同的RNA碱基对中间所造成的。表1.供试化合物与16S/18SrRNA结合作用试验结果<table>tableseeoriginaldocumentpage27</column></row><table><table>tableseeoriginaldocumentpage28</column></row><table>(2)采用表面等离子共振技术(surfaceplasmonresonance,SPR)测定了供试化合物与HIVTARRNA的结合能力,计算出供试化合物与RNA的解离常数(dissociationconstants,,^M),试验结果如表2所示。供试化合物与16SrRNA禾卩18SrRNA的结合与新霉素接近,提示供试化合物中方向环可以代替新霉素中的C、D环与RNA结合,具有潜在的抗艾滋病活性。表2.供试化合物与TARRNA结合作用试验结果<table>tableseeoriginaldocumentpage28</column></row><table>试验例l本发明化合物的抑菌活性试验1、供试化合物本发明实施例所制备的化合物,分别编号为9a,9b,9c,9d,10a,10b,10c,10d,11a,lib,llc,lid;2、试验方法及结果体外评价了供试化合物的对Raeruginosa标准株的抗菌活性,以新霉胺为阳性对照。结果如表3所示。测定化合物浓度为500网/mL时的抑菌率。试验结果显示,供试化合物均有抑菌活性;本发明对于抑菌活性好的化合物,进一步测定了其IC5Q。结果显示,在所有的供试化合物中,化合物9d抗菌最强。化合物9d和10d活性分别比化合物9a和10a强,推测可能易曲绕的长连接链有助于提高抗菌活性。表3供试化合物体外抑菌活性(P.aerag/"cwastrain)化合物IC50Og/mL)抑菌率*312.59b29%9c29%9d85.410a30%10b31%10c31%10d145.8lla30%lib31%lie30%lid18%Neaminc343.8权利要求1、新霉胺-咔啉羧酸缀合物,其结构式为通式I或通式II所示通式I通式II其中,其中,R选自氢或CH3O;R`选自氢、甲基或CH2CH(OCH3)2;X选自CONH,NHCO,NH,CO或氧;Y为氧,CH2,NH或不存在;Z为氧,CH2或NH;n=1,2,3,4,5或6。2、按照权利要求l所述的新霉胺咔啉羧酸缀合物,其特征在于R选自氢;R'选自氢或CH3;X为CONH;Y为不存在;Z为NH;n=l,2,3或4。3、一种制备权利要求l所述通式I化合物的方法,包括以下步骤(1)将新霉胺分别进行叠氮化反应和苯甲酰化反应得到0-[3',4'-二苯甲酰基-2',6'-二叠氮基-r,6、-二脱氧-。-0-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-1,3-二叠氮基-l,3-二脱氨-2-脱氧一D-链霉胺;将0-[3',4、-二苯甲酰基-2、,6、-二叠氮基-2、,6'-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-l,3-二叠氮基-l,3-二脱氨-2-脱氧一D-链霉胺游离羟基先三氟甲磺酸酯化,接着将三氟甲磺酸酯与相应的二胺氨解反应分别得到0-[3',4、-二苯甲酰基-2、,6'-二叠氮基-2、,6、-二脱氧-。-0-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5"(2-氨基乙氨基)-1,3-二叠氮基-l,3-二脱氨-2,5-二脱氧一D-链霉胺,0-[3、,4、-二苯甲酰基-2、,6'-二叠氮基-2',6'-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-(4-氨基丁氨基)1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺,0-[3',4'-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-(6-氨基己氨基)-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺,0-[3',4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5"(8-氨基癸氨基)-1,3-二叠氮基l,3-二脱氨-2,5-二脱氧一D-链霉胺化合物;(2)将步骤(1)所得到的化合物分别与3S-2-叔丁氧羰酰基-l,2,3,4-四氢-0-咔啉-3-羧酸进行縮合反应,得到的中间体在室温脱除Boc保护,分别得到0-[3、,4'-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基.(1—4)]-6-苯甲酰基-5a-[(2-(GS-2-叔丁氧羰酰基-1,2,3,小四氢-e-咔啉-3-基)-羰酰氨基)乙基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺;0-[3、,(-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5cc-[(小((3S-2-叔丁氧羰酰基-1,2,3,小四氢-e-咔啉-3-基)-羰酰氨基)丁基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一0-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(6-((35-2-叔丁氧羰酰基-1,2,3,4-四氢-P-咔啉-3-基)-羰酰氨基)己基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧—D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-^-0-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(8-((3S-2-叔丁氧羰酰基-1,2,3,小四氢-0-咔啉-3-基)-羰酰氨基)癸基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺;(3)将步骤(2)所得到的化合物在甲醇钠的甲醇溶液中皂化,随后还原叠氮基,即得。4、一种制备权利要求1述通式II化合物的方法,包括以下步骤(1)将新霉胺分别进行叠氮反应和苯甲酰化反应得到0-[3',4、-二苯甲酰基-2',6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-l,3-二叠氮基-1,3-二脱氨-2-脱氧-D-链霉胺;将0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-l,3-二叠氮基-l,3-二脱氨-2-脱氧一D-链霉胺游离羟基先三氟甲磺酸酯化,接着将三氟甲磺酸酯与相应的二胺氨解反应分别得到0-[3、,4'-二苯甲酰基-2',6、-二叠氮基-2、,6、-二脱氧-a-D-卩比喃葡萄糖基-(1—4)]-6-苯甲酰基-5"(2-氨基乙氨基)1,3-二叠氮基-l,3-二脱氨-2,5-二脱氧一D-链霉胺,0-[3、,4'-二苯甲酰基-2、,6'-二叠氮基-2',6'-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-(4-氨基丁氨基)1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺,0-[3',4'-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6_苯甲酰基-5a-(6-氨基己氨基)1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺,0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5"(8-氨基癸氨基)1,3-二叠氮基-l,3-二脱氨-2,5-二脱氧一D-链霉胺;(2)将步骤(1)所制备得到的化合物分别与e-咔啉-3-羧酸和i-甲基-e-咔啉-3-羧酸进行縮合,分别得到以下化合物0-[3、,4'-二苯甲酰基-2、,6、-二叠氮基-2、,6'-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(2-((-咔啉-3-基)-羰酰氨基)乙基)氨基]-l,3-二叠氮基-l,3-二脱氨-2,5-二脱氧一D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6'-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(4-((P-咔啉-3-基)-羰酰氨基)丁基)氨基]-l,3-二叠氮基-l,3-二脱氨-2,5-二脱氧一D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(6-((e-咔啉-3-基)-羰酰氨基)己基)氨基]-l,3-二叠氮基-l,3-二脱氨-2,5-二脱氧一D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6'-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(8-((e-咔啉-3-基)-羰酰氨基)癸基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺;0-[3',4'-二苯甲酰基-2、,6'-二叠氮基-2、,6'-二脱氧-(1-0-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a國[(2-((1-甲基-e-咔啉-3-基)-羰酰氨基)乙基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺;0-[3',4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-(:-0-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(4-((1-甲基-e-咔啉-3-基)-羰酰氨基)丁基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧一D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-a-D-吡喃葡萄糖基-(1—4)]-6-苯甲酰基-5a-[(6-((1-甲基-e-咔啉-3-基)-羰酰氨基)己基)氨基]-1,3-二叠氮基-1,3-脱氨-2,5-二脱氧—D-链霉胺;0-[3、,4、-二苯甲酰基-2、,6、-二叠氮基-2、,6、-二脱氧-。-0醫吡喃葡萄糖基-(1—4)〗-6-苯甲酰基-5a-[(8-((1-甲基-e隱咔啉-3-基)-羰酰氨基)癸基)氨基]-1,3-二叠氮基-1,3-二脱氨-2,5-二脱氧—D-链霉胺;(3)将步骤(2)所制备得到的化合物在甲醇钠的甲醇溶液中皂化,随后还原叠氮基,即得。5、按照权利要求4所述的方法,其特征在于步骤(2)该縮合反应以HOBT/TBTU做縮合剂,在DIPEA存在下依次进行缩合反应。6、一种抑菌药物组合物,其特征在于由治疗或预防上有效量的权利要求l所述的新霉胺-咔啉羧酸缀合物和药学上可接受的载体或辅料所组成。7、一种治疗艾滋病的药物组合物,其特征在于由治疗或预防上有效量的权利要求l所述的新霉胺-咔啉羧酸缀合物和药学上可接受的载体或辅料所组成。8、权利要求1所述的新霉胺-咔啉羧酸缀合物在制备抑菌药物中的用途。9、权利要求1所述的新霉胺-咔啉羧酸缀合物在制备抗艾滋病药物中的用途。全文摘要本发明公开了新霉胺-咔啉羧酸缀合物及其制备方法和在药学中的用途。本发明采用咔啉羧酸及其衍生物作为平面芳稠环结构,通过连接臂与新霉胺缀合,得到本发明新霉胺-咔啉羧酸缀合物。咔啉环通过嵌插作用,新霉胺通过静电、氢键作用,两者作用相辅相成,所获得的缀合物具有协同增效作用,可增强与靶标RNA的结合能力。体外抑菌试验结果表明,本发明缀合物均具有优良的抑菌活性,可作为临床潜在的抗菌制剂;本发明采用表面等离子共振技术测定了本发明缀合物与HIVTARRNA的结合能力,试验结果表明,本发明缀合物具有抗艾滋病病毒潜力,可作为临床潜在的抗艾滋病制剂。文档编号A61P31/18GK101580526SQ200910085159公开日2009年11月18日申请日期2009年6月2日优先权日2009年6月2日发明者叶新山,珊吴,吴艳芬申请人:北京大学
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