一种平滑受体配体

文档序号：1433989 发布日期：2020-03-20 浏览：32次 >En<

阅读说明：本技术 一种平滑受体配体 (Smooth receptor ligand ) 是由陶厚朝赵飞于 2018-09-13 设计创作，主要内容包括：本发明涉及生物技术领域,特别是涉及一种平滑受体配体。本发明提供一种平滑受体配体或其异构体前体药物、溶剂化物、药学上可接受的盐,所述平滑受体配体的结构式为：A——linker——B其中,A为胞外域配体结构,B为跨膜域配体结构；Linker为线性的、对平滑受体无活性的亚基。本发明所提供的针对平滑受体的新型双头小分子配体,结合平滑受体的晶体结构数据,在胞外域配体和跨膜域配体的适当位点引入连接体,得到全新的双头配体小分子,增强了配体和受体之间的相互作用以及配体的生物活性。(The invention relates to the field of biotechnology, in particular to a smooth receptor ligand. The invention provides a smooth receptor ligand or an isomer prodrug, solvate and pharmaceutically acceptable salt thereof, wherein the structural formula of the smooth receptor ligand is as follows: A-linker-B, wherein A is an extracellular domain ligand structure, and B is a transmembrane domain ligand structure; linker is a linear, smooth receptor inactive subunit. The novel double-headed small molecule ligand for the smooth receptor provided by the invention is combined with the crystal structure data of the smooth receptor, and a connector is introduced into a proper site of an ectodomain ligand and a transmembrane domain ligand to obtain a brand-new double-headed ligand small molecule, so that the interaction between the ligand and the receptor and the biological activity of the ligand are enhanced.)

一种平滑受体配体

技术领域

本发明涉及生物技术领域，特别是涉及一种平滑受体配体。

背景技术

平滑受体属于G-蛋白偶联受体(G protein-coupled receptors，GPCRs)中的F家族，其功能紊乱将导致出生缺陷。20世纪50年代在美国西部发现许多出生小羊只有一只眼睛，通过对此种独眼症的研究发现了平滑受体的第一个天然植物小分子配体环巴胺(Cyclopamine)。平滑受体的功能调控与一系列癌症的发生发展相关，因此平滑受体是医药开发中的重要靶标。在过去的几十年中，许多的先导化合物配体被发现，其中两个小分子药物在2015年被推向市场。然而，平滑受体突变体导致的抗药性很快显现出来，亟需针对现有药物进行进一步的改造。

发明内容

鉴于以上所述现有技术的缺点，本发明的目的在于提供一种平滑受体配体，用于解决现有技术中的问题。

为实现上述目的及其他相关目的，本发明一方面提供一种平滑受体配体或其异构体前体药物、溶剂化物、药学上可接受的盐，所述平滑受体配体的结构式为：

A-linker-B

其中，A为胞外域配体结构，B为跨膜域配体结构；

Linker为线性的、对平滑受体无活性的亚基。

在本发明一些实施方式中，所述胞外域配体结构为甾体结构。

在本发明一些实施方式中，A选自如下所示的基团：

其中，E选自H，-OH；

F选自含有或不含有选自N、S、P、O杂原子的直链的C1～C20脂肪族基团；

G选自含有或不含有选自N、S、P、O杂原子的直链的C1～C20烷基或酰基。

在本发明一些实施方式中，A的结构选自20-(S)-羟基胆固醇、22-(S)-氮杂胆固醇、环巴胺、胆固醇、石胆酸、20-(S)-羟基-24-炔基胆固醇。

在本发明一些实施方式中，A选自如下所示的基团：

在本发明一些实施方式中，所述跨膜域配体结构可以是含有多个脂肪环、含氮脂肪环、苯环、氮杂芳香环系的酰胺或亚胺类结构。

在本发明一些实施方式中，B选自如下所示的基团：

其中，J选自取代基各自独立地选自烷基、烯基、炔基、F、Cl、Br、I、硝基、氨基、羟基、烷氧基、环烷基、芳基、杂芳基的取代或未取代的苯基、取代或未取代的苄基、取代或未取代的吡啶基、取代或未取代的环烷基；

L选自氢、烷基、取代基选自-NR’R”取代或未取代的环烷基，其中，R’和R”各自独立地选自C1～C6烷基；

M选自取代基选自F、Cl、Br、I、C1～C6烷基、C1～C6烷氧基、-CF₃、-OCF₃、烷基磺酰基的取代或未取代的芳基或杂芳基。

在本发明一些实施方式中，B的结构选自SAG、LY2940680、GDC0449、LDE225、SANT1、SANT2。

在本发明一些实施方式中，B选自如下所示的基团：

在本发明一些实施方式中，所述连接体为具有***结构的聚乙二醇低聚物。在本发明一些实施方式中，所述连接体的结构式如下所示：

其中m，n各自独立地选自0～7之间的整数。

在本发明一些实施方式中，所述平滑受体配体选自如下所示的化合物：

本发明另一方面提供所述的平滑受体配体或其异构体、前体药物、溶剂化物、药学上可接受的盐在制备平滑受体激动剂、抑制剂或拮抗剂中的用途。

本发明另一方面提供一种药物组合物，包括所述的平滑受体配体或其异构体、前体药物、溶剂化物、药学上可接受的盐

附图说明

图1显示为本发明通过晶体结构确认胞外域配体与跨膜域配体的分子取向及间距示意图。

图2显示为本发明双头分子同时结合平滑受体两个结构域示意图。

图3显示为本发明一系列双头分子的活性与连接体长度之间表现出U形曲线关系。

具体实施方式

本发明发明人将平滑受体的各结合域的受体通过适当的连接体整合在一起，通过邻近效应可实现小分子药物同时结合两个结构域，从而增强药物活性，并实现规避抗药性的小分子药物设计，在此基础上完成了本发明。

本发明一方面提供一种平滑受体配体或其异构体(例如，对映异构体、非对映异构体、几何异构体、互变异构体、旋转异构体、阻转异构体、消旋体等)、前体药物、溶剂化物、药学上可接受的盐，所述平滑受体配体的结构式为：

A-linker-B

其中，A为胞外域配体结构，B为跨膜域配体结构；

Linker(连接体)为线性的、对平滑受体无活性的亚基。

本发明所提供的平滑受体配体中，A为胞外域配体结构，所述胞外域配体结构通常指能够对平滑受体的胞外域具有识别能力并与之结合的分子结构，所述识别和结合可以是特异性的。所述胞外域配体结构可以是抑制剂结构、拮抗剂结构或激动剂结构。所述胞外域配体结构可以是甾体结构，例如，A可以选自如下所示的基团：

其中，E选自H(氢原子)，-OH(羟基)；F选自含有或不含有杂原子的C1～C20、 C1～C12、C1～C6、C1～C3直链的脂肪族基团，基团F中，杂原子可以是N、S、P、O等；G 选自含有或不含有杂原子的C1～C20、C1～C12、C1～C6、C1～C3直链烷基或酰基，基团G 中，杂原子可以是N、S、P、O等。更具体的，所述胞外域配体结构可以是包括但不限于 20-(S)-羟基胆固醇、22-(S)-氮杂胆固醇、环巴胺、胆固醇、石胆酸、20-(S)-羟基-24-炔基胆固醇以及其它甾体所对应的基团结构等。在本发明另一些具体实施方式中，A可以选自如下所示的基团：

本发明所提供的平滑受体配体中，B为跨膜域配体结构，所述跨膜域配体结构通常指能够对平滑受体的跨膜域具有识别能力并与之结合的分子结构，所述识别和结合可以是特异性的。所述跨膜域配体结构可以是抑制剂结构、拮抗剂结构或激动剂结构。所述跨膜域配体结构可以是含有多个脂肪环、含氮脂肪环、苯环、氮杂芳香环系的酰胺或亚胺类结构，例如， B可以选自如下所示的基团：

其中，J选自取代或未取代的苯基，取代或未取代的苄基，取代或未取代的吡啶基，取代或未取代的C3～C9环烷基，基团J中，苯基、苄基、吡啶基、环烷基可以是单取代的，也可以是多取代的，它们的取代基各自独立地选自C1～C20、C1～C12、C1～C6、C1～C3脂肪族基团、F、Cl、Br、I等卤原子、硝基、氨基、羟基、烷氧基、环烷基、C1～C20、C1～C12、 C1～C6芳基、C1～C20、C1～C12、C1～C6杂芳基；

L选自氢，C1～C20、C1～C12、C1～C6、C1～C3烷基，基团L中，烷基可以是单取代的，也可以是多取代的，它们的取代基选自-NR’R”取代或未取代的C3～C9环烷基，其中， R’和R”各自独立地选自C1～C6、C1～C3烷基；

M选自C1～C20、C1～C12、C1～C6芳基或C1～C20、C1～C12、C1～C6杂芳基，基团M中，芳基或杂芳基可以是单取代的，也可以是多取代的，它们的取代基选自F、Cl、Br、I等卤原子、C1～C6烷基、C1～C6烷氧基、-CF₃、-OCF₃、烷基磺酰基。更具体的，可以是包括但不限于SAG(CAS号：912545-86-9)、LY2940680(CAS号：1258861-20-9)、GDC0449 (CAS号：879085-55-9)、LDE225(CAS号：956697-53-3)、SANT1(CAS号：304909-07-7)、 SANT2(CAS号：329196-48-7)等所对应的基团结构。在本发明另一些具体实施方式中，B 可以选自如下所示的基团：

本发明中，所述脂肪族基团通常指链状烃类化合物，例如可以是烷基、烯基和炔基。所述脂肪族基团可以是包括但不限于甲基、乙基、乙烯基、乙炔基、丙基、丙烯基、丙炔基、丁基、丁烯基、丁炔基、戊基、己基、庚基、辛基、壬基和癸基等。脂肪族基团中可以包含杂原子，例如，杂原子可以是N、S、P、O等。

本发明中，所述环烷基应被理解为是指饱和的与不饱和的(但不是芳族的)环状烃，包括但不限于环丙基、环丁基、环戊基、环己基、环庚基等。对于环烷基，还包括饱和的环烷基，其中任选地至少一个碳原子可以被杂原子替换，优选的杂原子可以是S、N、P或O 等。另外，对于环中没有杂原子的单不饱和或多不饱和(优选单不饱和)的环烷基，只要其不是芳香族体系，也属于环烷基。

本发明中，所述烷氧基包括但不限于甲氧基、乙氧基、丙氧基等。所述烷氧基可以是单取代的，也可以是多取代的，所述烷氧基的取代基包括但不限于F、Cl、Br等卤原子等，所述取代的烷氧基可以是包括但不限于氟代丙氧基、三氟代乙氧基等。

本发明中，所述芳基通常指具有至少一个芳香环的闭环体系，通常不包括杂原子(例如，N、O等。所述芳基包括但不限于苯基、萘基、荧蒽基、芴基、四氢化萘基、茚满基、蒽基、菲基、二氢菲基等。

本发明中，杂芳基应理解为是指杂环体系，其具有至少一个芳香环，并且任选地含有选自N、O、S等中的一个或多个的杂原子，所述杂芳环基包括但不限于呋喃基、噻吩基、吡咯基、吡唑基、噁唑基、噻唑基、***基、吡啶基、哒嗪基、嘧啶基、吡嗪基、三嗪基、苯并呋喃基、苯并噻吩基、吲哚基、异吲哚基、苯并咪唑基、吲唑基、苯并噻唑基、苯并噁唑基、喹啉基、异喹啉基、喹喔啉基、呔嗪基、喹唑啉基、噌啉基、嘌呤基、咔唑基、二苯并呋喃基、吖啶基、吩嗪基、吩噁嗪、吩噻嗪、吩噁噻等。

本发明所提供的平滑受体配体中，所述连接体通常可以是线性的，还可以是对平滑受体无活性。连接体的长度对于配体的活性强弱有影响，所述连接体通常可以是具有***结构的聚乙二醇低聚物(PEG oligomer with triazole)，在本发明一些具体实施方式中，linker的结构式如下所示：

其中m，n各自独立地选自0～7之间的整数，例如，m可以是0、1、2、3、4、5、6或 7，再例如，n可以是0、1、2、3、4、5、6或7。

在本发明一些具体实施方式中，所述平滑受体配体可以具体选自结构式如下的化合物：

本发明另一方面提供所述平滑受体配体或其异构体、前体药物、溶剂化物、药学上可接受的盐在制备平滑受体(SMO受体，smoothened receptor)激动剂、抑制剂或拮抗剂中的用途。所述激动剂通常指能够促进某个分子活性、某种反应或者某个通路的一类物质，所述抑制剂通常指能够降低或阻滞某个分子活性、某种反应或者某个通路的一类物质。所述拮抗剂通常指与受体结合后阻断另一物质的激动剂介导的作用，且本身不引起该物质所对应的生物学效应的一类物质。本发明所提供的平滑受体配体或其异构体、前体药物、溶剂化物、药学上可接受的盐，当连接体长度在一定范围内时，连接体优化后的双头分子是以同时结合平滑受体的两个结合域的模式作用于受体，通过协同效应达到了更高的活性，所达到的活性种类通常与配体的活性种类相关，例如，所达到的活性效果可以是激动、抑制、拮抗等作用。

本发明另一方面提供一种药物组合物，包括如上所述的平滑受体配体或其异构体、前体药物、溶剂化物、药学上可接受的盐。所述药物组合物中，平滑受体配体或其异构体、前体药物、溶剂化物、药学上可接受的盐可以是唯一药效成分，也可以与其他至少一种或多种有效成分组合，联合用药。

本发明所提供的药物组合物中，还可以包括药学上可接受的辅料或添加剂，所述辅料或添加剂可以选自载体、赋形剂、支撑材料、助剂、润滑剂、填充剂、溶剂、稀释剂、着色剂、调味剂等。

本发明所述的药物组合物可以适应于以任何形式给药，可以是口服或胃肠外给药，具体可以是经肺、经鼻、经直肠和/或静脉注射等，再例如，可以适于局部或全身应用，具体可以是真皮内、皮下、肌内、关节内、腹膜内、肺部、口腔、舌下含服、经鼻、经皮、***、口服或胃肠外应用等。

本发明所提供的针对平滑受体的新型双头小分子配体，结合平滑受体的晶体结构数据，在胞外域配体和跨膜域配体的适当位点引入连接体，得到全新的双头配体小分子，增强了配体和受体之间的相互作用以及配体的生物活性。此类小分子配体可以作为工具分子应用于平滑受体的功能和结构研究，也可以作为与平滑受体相关疾病的医药开发候选。

以下通过特定的具体实例说明本发明的实施方式，本领域技术人员可由本说明书所揭露的内容轻易地了解本发明的其他优点与功效。本发明还可以通过另外不同的具体实施方式加以实施或应用，本说明书中的各项细节也可以基于不同观点与应用，在没有背离本发明的精神下进行各种修饰或改变。

在进一步描述本发明具体实施方式之前，应理解，本发明的保护范围不局限于下述特定的具体实施方案；还应当理解，本发明实施例中使用的术语是为了描述特定的具体实施方案，而不是为了限制本发明的保护范围；在本发明说明书和权利要求书中，除非文中另外明确指出，单数形式“一个”、“一”和“这个”包括复数形式。

当实施例给出数值范围时，应理解，除非本发明另有说明，每个数值范围的两个端点以及两个端点之间任何一个数值均可选用。除非另外定义，本发明中使用的所有技术和科学术语与本技术领域技术人员通常理解的意义相同。除实施例中使用的具体方法、设备、材料外，根据本技术领域的技术人员对现有技术的掌握及本发明的记载，还可以使用与本发明实施例中所述的方法、设备、材料相似或等同的现有技术的任何方法、设备和材料来实现本发明。

除非另外说明，本发明中所公开的实验方法、检测方法、制备方法均采用本技术领域常规的分子生物学、生物化学、染色质结构和分析、分析化学、细胞培养、重组DNA技术及相关领域的常规技术。这些技术在现有文献中已有完善说明，具体可参见Sambrook等MOLECULAR CLONING：A LABORATORY MANUAL，Second edition，Cold Spring HarborLaboratory Press，1989 and Third edition，2001；Ausubel等，CURRENT PROTOCOLS INMOLECULAR BIOLOGY，John Wiley&Sons，New York，1987 and periodic updates；theseries METHODS IN ENZYMOLOGY，Academic Press，San Diego； Wolffe，CHROMATINSTRUCTURE AND FUNCTION，Third edition，Academic Press，San Diego，1998；METHODS INENZYMOLOGY，Vol.304，Chromatin(P.M.Wassarman and A.P.Wolffe，eds.)，AcademicPress，San Diego，1999；和METHODS IN MOLECULAR BIOLOGY，Vol.119，ChromatinProtocols(P.B.Becker，ed.)Humana Press，Totowa，1999 等。

实施例1

配体及其合成：

平滑受体的双头配体，其结构式分别为(I)-(XXIX)。

所有双头配体均由一个末端连接有乙炔基的中间体及一个末端连接有叠氮基的中间体在一价铜盐的催化下反应得到。双头配体的合成通式如下：

：通过Click反应合成平滑受体的双头配体。反应条件：(a)CuSO₄,sodiumascorbate, MeOH,r.t.,1h.

中间体结构及编号对应如下：

片段分子合成方法如下：

如上所示的反应方程式为TC577及其同系物的合成路线，反应条件具体如下：

(a)TBSCl,imidazole,DMAP,DMF/THF,r.t.,1h；

(b)6-bromo-1-hexene,Mg,THF,r.t.,3h；

(c)i.9-BBN,THF,r.t.,1h；ii.1M NaOH,H₂O₂,0℃ to r.t.,1h；

(d)3-bromo-1-propyne,NaH(60％),THF,50℃,2h；

(e)TBAF,THF,r.t.,1h；

(f)TsCl,Et₃N,DMAP,DCM,r.t.,1h；

(g)ethylene glycol,NaH(60％),DMF,70℃,6h；

(h)1-propargyl oligo ethylene glycol,NaH(60％),THF,reflux,4-6h.

各步骤具体的制备方法如下：

20.0g孕烯醇酮、18.0g咪唑及769mg 4-二甲氨基吡啶(DMAP)溶于30mL N,N-二甲基甲酰胺(DMF)和65mL四氢呋喃(THF)的混合溶剂中，加入18.0g叔丁基二甲基氯硅烷(TBSCl)，室温搅拌1小时。反应结束后加入20mL饱和氯化铵溶液淬灭，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品在100mL正己烷和 10mL二氯甲烷的混合溶剂中通过热重结晶得到大部分纯净的无色晶状中间体1。母液用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝10:1)分离得到纯净的中间体1。

在干燥的、充满氮气的反应瓶中加入28.8g镁屑。将10.0g 6-溴-1-己烯溶于60mL无水 THF中，该溶液的约1/10加至镁屑中，加入1滴1,2-二溴乙烷并适当加热引发格氏试剂的形成。引发成功后将剩余的溴代物溶液滴加至镁屑中，控制滴加速率以使反应液保持微沸。滴加完毕后继续加热回流2小时。将反应液冷却，使用单质碘标定格氏试剂的浓度。一般地，该方法合成得到的格氏试剂浓度约为0.5-0.7M。

在干燥的、充满氮气的反应瓶中将5.0g中间体1溶于35mL无水THF中，向此溶液滴加65mL,0.54M的1-(5-己烯基)溴化镁的THF溶液，室温搅拌3小时。反应结束后加入50 mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝3:1)分离得到纯净的中间体2。

在干燥的、充满氮气的反应瓶中将2.0g中间体2溶于40mL无水THF中，向此溶液滴加0.5M 9-硼双环[3.3.1]壬烷(9-BBN)的THF溶液40mL，室温搅拌1小时。之后将反应降温至0℃，缓慢滴加20mL1M氢氧化钠溶液及20mL 30％过氧化氢，0℃下继续搅拌1小时。反应结束后加入20mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝1:1)分离得到纯净的中间体3。¹H NMR(500MHz,CDCl₃),(ppm)0.06(s,6H,CH₃), 0.86(s,3H,CH₃),0.89(s,9H,CH₃),0.90-0.98(m,1H,CH₂),1.00(s,3H,CH₃),1.01-1.25(m,4H, CH₂andCH),1.26(s,3H,CH₃),1.29-1.76(m,21H,CH₂and CH),1.78-1.82(m,1H,CH₂), 1.94-2.00(m,1H,CH₂),2.05-2.10(m,1H,CH₂),2.14-2.19(m,1H,CH₂),2.24-2.30(m,1H,CH₂), 3.46-3.50(m,1H,CH),3.63-3.65(t,J＝8.5Hz,2H,CH₂),4.21(d,J＝2.0Hz,2H,CH₂),5.31-5.32(m,1H,CH)；¹³C NMR(125MHz,CDCl₃)δ(ppm)-4.6,13.6,18.3,19.4,20.9,22.3,23.8,24.2,25.7,25.9,26.4,30.0,31.3,31.8,32.0,32.7,36.6,37.3,40.1,42.6,42.8,43.8,50.1,56.9,57.6,63.0, 72.6,75.2,121.0,141.6.

在106mg中间体3的2mL THF溶液中加入16mg NaH(60％)，室温搅拌15分钟，然后加入24mg 3-溴丙炔，加热至50℃反应2小时。反应结束后加入5mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝3:1)分离得到纯净的中间体4。¹H NMR(500MHz,CDCl₃),(ppm)0.06(s,6H,CH₃),0.86(s,3H,CH₃),0.89(s,9H,CH₃), 0.91-0.98(m,1H,CH₂),1.00(s,3H,CH₃),1.01-1.22(m,4H,CH₂and CH),1.26(s,3H,CH₃), 1.29-1.76(m,21H,CH₂and CH),1.78-1.82(m,1H,CH₂),1.94-1.99(m,1H,CH₂),2.06-2.10(m, 1H,CH₂),2.15-2.19(m,1H,CH₂),2.24-2.29(m,1H,CH₂),2.42(t,J＝2.0Hz,1H,CH),3.46-3.52 (m,3H,CH₂and CH),4.13(d,J＝2.5Hz,2H,CH₂),5.31-5.32(m,1H,CH)；¹³C NMR(125MHz, CDCl₃),-4.6,13.6,18.2,19.4,20.8,22.3,23.7,24.2,25.9,26.0,26.4,29.4,30.0,31.3,31.8,32.0,36.5,37.3,40.1,42.6,42.7,43.9,50.0,56.9,57.5,58.0,70.2,72.6,74.0,75.1,80.0,121.0, 141.5.

在57mg中间体4的1mL THF溶液中加入1M四丁基氟化铵(TBAF)的THF溶液0.5 mL，室温搅拌1小时。反应结束后浓缩、用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝1:1)分离得到纯净的中间体TC577。¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H, CH₃),0.88-0.99(m,1H,CH₂),1.01(s,3H,CH₃),1.02-1.25(m,4H,CH₂and CH),1.26(s,3H, CH₃),1.27-1.78(m,20H,CH₂and CH),1.82-1.86(m,2H,CH₂),1.95-2.00(m,1H,CH₂),2.07-2.11 (m,1H,CH₂),2.20-2.32(m,2H,CH₂),2.43(t,J＝2.0Hz,1H,CH),3.49-3.54(m,3H,CH₂and CH),4.13(d,J＝2.5Hz,2H,CH₂),5.35-5.36(m,1H,CH)；¹³C NMR(125MHz,CDCl₃), 13.6,19.4,20.9,22.3,23.7,24.2,26.0,26.4,29.4,30.0,31.2,31.6,31.7,36.4,37.2,40.1,42.2,42.6,43.9,49.9,56.8,57.5,58.0,70.2,71.6,74.0,75.1,80.0,121.5,140.7.

5.32g中间体3、2.02g三乙胺及122mg DMAP溶于50mL二氯甲烷(DCM)中，加入2.85g对甲苯磺酰氯(TsCl)，室温搅拌1小时。反应结束后加入40mL饱和碳酸氢钠溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和碳酸氢钠水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝7:1)分离得到纯净的中间体5。¹H NMR(500MHz,CDCl₃),(ppm)0.08(s,6H,CH₃),0.87(s,3H,CH₃),0.90(s,9H,CH₃), 0.90-0.98(m,1H,CH₂),0.99(s,3H,CH₃),1.01-1.25(m,4H,CH₂and CH),1.24(s,3H,CH₃), 1.29-1.76(m,21H,CH₂and CH),1.78-1.84(m,1H,CH₂),1.93-1.98(m,1H,CH₂),2.05-2.08(m, 1H,CH₂),2.14-2.17(m,1H,CH₂),2.22-2.27(m,1H,CH₂),2.43(s,3H,CH₃),3.43-3.53(m,1H, CH),4.00(t,J＝6.5Hz,2H,CH₂),5.29-5.32(m,1H,CH),7.33(d,J＝8.0Hz,2H,CH),7.77(d,J ＝8.0Hz,2H,CH)；¹³C NMR(125MHz,CDCl₃)δ(ppm)-3.6,13.6,18.2,19.4,20.9,21.6,22.3,23.7,24.0,25.3,25.6,25.9,26.3,28.8,29.5,31.3,31.6,31.8,32.0,32.7,36.5,37.2,40.1,42.2,42.6, 43.7,50.0,56.8,57.6,70.5,71.7,72.5,75.1,120.1,121.5,127.8,129.8,133.2,140.7,141.5,144.6.

在542mg乙二醇和5mL DMF的混合溶液中加入146mg NaH(60％)，室温搅拌15 分钟，然后加入501mg中间体5，加热至70℃反应4-6小时。反应结束后加入20mL饱和氯化铵溶液淬灭反应，用二氯甲烷萃取三次，有机相依次用饱和碳酸氢钠水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析 (200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝2:1)分离得到纯净的中间体6。

在117mg中间体6的2mL THF溶液中加入16mg NaH(60％)，室温搅拌15分钟，然后加入24mg 3-溴丙炔，加热至50℃反应2小时。反应结束后加入5mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝3:1)分离得到纯净的中间体7。¹H NMR(500MHz,CDCl₃),(ppm)0.06(s,6H,CH₃),0.86(s,3H,CH₃),0.89(s,9H,CH₃), 0.90-0.98(m,1H,CH₂),1.00(s,3H,CH₃),1.01-1.25(m,4H,CH₂and CH),1.26(s,3H,CH₃), 1.29-1.76(m,21H,CH₂and CH),1.78-1.82(m,1H,CH₂),1.94-2.00(m,1H,CH₂),2.05-2.10(m, 1H,CH₂),2.15-2.19(m,1H,CH₂),2.24-2.30(m,1H,CH₂),2.44(t,J＝2.5Hz,1H,CH),3.44-3.51 (m,3H,CH₂and CH),3.60-3.62(m,2H,CH₂),3.69-3.71(m,2H,CH₂),4.21(d,J＝2.0Hz,2H, CH₂),5.31-5.32(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),-4.6,13.6,18.2,19.4,20.8,22.3, 23.7,24.2,25.9,26.0,26.4,29.5,30.0,31.2,31.7,32.0,36.5,37.3,40.0,42.6,42.7,43.9,50.0,56.8, 57.5,58.4,69.1,69.8,71.5,72.5,74.5,75.1,79.6,121.0,141.5.HRMScalcd for C₃₈H₆₆O₄Si [M+Na]⁺:637.4623；found:637.4635.

在61mg中间体7的1mL THF溶液中加入1M四丁基氟化铵(TBAF)的THF溶液0.5 mL，室温搅拌1小时。反应结束后浓缩、用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝1:1)分离得到纯净的中间体TC573。¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H, CH₃),0.88-0.99(m,1H,CH₂),1.01(s,3H,CH₃),1.02-1.25(m,4H,CH₂and CH),1.26(s,3H, CH₃),1.27-1.76(m,20H,CH₂and CH),1.81-1.86(m,2H,CH₂),1.95-1.99(m,1H,CH₂),2.06-2.10 (m,1H,CH₂),2.21-2.31(m,2H,CH₂),2.44(t,J＝2.5Hz,1H,CH),3.44-3.54(m,3H,CH₂and CH),3.60-3.62(m,2H,CH₂),3.69-3.71(m,2H,CH₂),4.21(d,J＝2.5Hz,2H,CH₂),5.34-5.35(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),13.6,19.3,20.8,22.2,23.7,24.2,26.0,26.3,29.5,30.0,31.2,31.5,31.7,36.4,37.1,40.0,42.2,42.5,43.8,49.9,56.8,57.4,58.3,69.0,69.8,71.4,71.6, 74.5,75.2,79.6,121.5,140.7；HRMS calcd for C₃₂H₅₂O₄[M+Na]⁺:523.3758；found:523.3757。

合成TC574及同系物的通用步骤(以TC574的合成为例)：

在58mg 1-炔丙基二乙二醇的2mL THF溶液中加入24mg NaH(60％)，室温搅拌15分钟，然后加入中间体5，加热回流6小时。反应结束后加入5mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析 (200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝5:1)分离得到纯净的中间体8。¹H NMR(500 MHz,CDCl₃),(ppm)0.06(s,6H,CH₃),0.86(s,3H,CH₃),0.89(s,9H,CH₃),0.90-0.98(m,1H, CH₂),1.00(s,3H,CH₃),1.01-1.25(m,4H,CH₂and CH),1.26(s,3H,CH₃),1.29-1.76(m,21H,CH₂and CH),1.78-1.82(m,1H,CH₂),1.95-1.99(m,1H,CH₂),2.06-2.10(m,1H,CH₂),2.15-2.19 (m,1H,CH₂),2.24-2.30(m,1H,CH₂),2.44(t,J＝2.5Hz,1H,CH),3.43-3.50(m,3H,CH₂and CH),3.58-3.73(m,8H,CH₂),4.21(d,J＝2.5Hz,2H,CH₂),5.31-5.32(m,1H,CH)；¹³CNMR(125 MHz,CDCl₃),-4.6,13.6,18.2,19.4,20.8,22.3,23.7,24.2,25.9,26.0,26.4,29.5,30.0,31.2, 31.7,32.0,36.5,37.3,40.0,42.6,42.7,43.9,50.0,56.8,57.5,58.4,69.0,70.0,70.4,70.6,71.4,72.5, 74.5,75.1,79.6,121.0,141.5；HRMS calcd forC₄₀H₇₀O₅Si[M+Na]⁺:681.4885；found:681.4898.

中间体9。¹H NMR(500MHz,CDCl₃),(ppm)0.06(s,6H,CH₃),0.85(s,3H,CH₃),0.89(s,9H,CH₃),0.90-0.98(m,1H,CH₂),1.00(s,3H,CH₃),1.01-1.23(m,4H,CH₂and CH),1.26(s, 3H,CH₃),1.29-1.76(m,21H,CH₂and CH),1.78-1.82(m,1H,CH₂),1.94-1.99(m,1H,CH₂), 2.06-2.10(m,1H,CH₂),2.14-2.18(m,1H,CH₂),2.24-2.29(m,1H,CH₂),2.45(t,J＝2.5Hz,1H, CH),3.43-3.50(m,3H,CH₂and CH),3.57-3.72(m,12H,CH₂),4.20(d,J＝2.5Hz,2H,CH₂), 5.31-5.32(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),-4.7,13.5,18.1,19.3,20.8,22.2,23.7, 24.1,25.8,25.9,26.3,29.4,30.0,31.2,31.6,31.9,36.4,37.2,40.0,42.5,42.6,43.8,49.9,56.8,57.4, 58.2,68.9,69.9,70.2,70.41,70.46,71.3,72.4,74.5,75.0,79.5,120.9,141.3；HRMS calcd for C₄₂H₇₄O₆Si[M+Na]⁺:725.5147；found:725.5145.

中间体10。¹H NMR(500MHz,CDCl₃),(ppm)0.06(s,6H,CH₃),0.86(s,3H,CH₃),0.89(s,9H,CH₃),0.90-0.98(m,1H,CH₂),1.00(s,3H,CH₃),1.01-1.22(m,4H,CH₂and CH),1.26(s, 3H,CH₃),1.28-1.76(m,21H,CH₂and CH),1.78-1.82(m,1H,CH₂),1.94-1.99(m,1H,CH₂), 2.06-2.10(m,1H,CH₂),2.14-2.19(m,1H,CH₂),2.24-2.30(m,1H,CH₂),2.44(t,J＝2.5Hz,1H, CH),3.43-3.51(m,3H,CH₂and CH),3.57-3.73(m,16H,CH₂),4.20(d,J＝2.5Hz,2H,CH₂), 5.31-5.32(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),-4.6,13.5,18.2,19.4,20.8,22.2,23.7, 24.2,25.9,26.0,26.4,29.5,30.0,31.2,31.7,32.0,36.5,37.3,40.0,42.5,42.7,43.9,50.0,56.8,57.4, 58.3,69.0,70.0,70.3,70.47,70.49,70.51,70.52,70.53,71.4,72.5,74.5,75.1,79.5,120.9,141.3； HRMS calcd for C₄₄H₇₈O₇Si[M+Na]⁺:769.5409；found:769.5413.

中间体11。¹H NMR(500MHz,CDCl₃),(ppm)0.06(s,6H,CH₃),0.85(s,3H,CH₃),0.89(s,9H,CH₃),0.90-0.98(m,1H,CH₂),1.00(s,3H,CH₃),1.01-1.23(m,4H,CH₂and CH),1.26(s, 3H,CH₃),1.29-1.76(m,21H,CH₂and CH),1.78-1.82(m,1H,CH₂),1.94-1.99(m,1H,CH₂), 2.06-2.10(m,1H,CH₂),2.14-2.18(m,1H,CH₂),2.24-2.29(m,1H,CH₂),2.45(t,J＝2.5Hz,1H, CH),3.43-3.50(m,3H,CH₂and CH),3.57-3.72(m,20H,CH₂),4.20(d,J＝2.5Hz,2H,CH₂), 5.31-5.32(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),-4.7,13.5,18.1,19.3,20.8,22.2,23.7, 24.1,25.8,25.9,26.3,29.4,30.0,31.2,31.6,31.9,36.4,37.2,40.0,42.5,42.6,43.8,49.9,56.8,57.4, 58.2,68.9,69.9,70.2,70.41,70.46,71.3,72.4,74.5,75.0,79.5,120.9,141.3；HRMS calcd for C₄₆H₈₂O₈Si[M+Na]⁺:813.5671；found:813.5670.

中间体12。¹H NMR(500MHz,CDCl₃),(ppm)0.06(s,6H,CH₃),0.86(s,3H,CH₃),0.89(s,9H,CH₃),0.90-0.98(m,1H,CH₂),1.00(s,3H,CH₃),1.01-1.22(m,4H,CH₂and CH),1.26(s, 3H,CH₃),1.28-1.76(m,21H,CH₂and CH),1.78-1.82(m,1H,CH₂),1.94-1.99(m,1H,CH₂), 2.06-2.10(m,1H,CH₂),2.14-2.19(m,1H,CH₂),2.24-2.30(m,1H,CH₂),2.44(t,J＝2.5Hz,1H, CH),3.43-3.51(m,3H,CH₂and CH),3.57-3.73(m,24H,CH₂),4.20(d,J＝2.5Hz,2H,CH₂), 5.31-5.32(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),(ppm)-4.6,13.5,18.2,19.4,20.8,22.2, 23.7,24.2,25.9,26.0,26.4,29.5,30.0,31.2,31.7,32.0,36.5,37.3,40.0,42.5,42.7,43.9,50.0,56.8, 57.4,58.3,69.0,70.0,70.3,70.47,70.49,70.51,70.52,70.53,71.4,72.5,74.5,75.1,79.5,120.9, 141.3；HRMS calcd for C₄₈H₈₆O₉Si[M+Na]⁺:857.5933；found:857.5923.

在65mg中间体8的1mL THF溶液中加入1M四丁基氟化铵(TBAF)的THF溶液0.5 mL，室温搅拌1小时。反应结束后浓缩、用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝1:1)分离得到纯净的中间体TC574。¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H, CH₃),0.88-0.99(m,1H,CH₂),1.01(s,3H,CH₃),1.02-1.24(m,4H,CH₂and CH),1.26(s,3H, CH₃),1.28-1.76(m,20H,CH₂and CH),1.81-1.86(m,2H,CH₂),1.95-2.01(m,1H,CH₂),2.07-2.10 (m,1H,CH₂),2.21-2.32(m,2H,CH₂),2.44(t,J＝2.5Hz,1H,CH),3.44-3.54(m,3H,CH₂and CH),3.58-3.60(m,2H,CH₂),3.63-3.72(m,6H,CH₂),4.21(d,J＝2.5Hz,2H,CH₂),5.34-5.35(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),13.6,19.3,20.8,22.2,23.7,24.2,26.0,26.3,29.5,30.0,31.2,31.5,31.7,36.4,37.2,40.0,42.2,42.5,43.9,49.9,56.8,57.4,58.3,69.0,70.0,70.3,70.6, 71.4,71.6,74.5,75.1,79.6,121.5,140.7；HRMS calcd for C₃₄H₅₆O₅[M+Na]⁺:567.4020；found: 567.4017。

TC575。¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H,CH₃),0.88-0.99(m,1H,CH₂),1.01(s,3H,CH₃),1.02-1.23(m,4H,CH₂and CH),1.26(s,3H,CH₃),1.28-1.76(m,20H,CH₂andCH),1.81-1.86(m,2H,CH₂),1.95-1.99(m,1H,CH₂),2.06-2.10(m,1H,CH₂),2.20-2.30(m,2H, CH₂),2.45(t,J＝2.5Hz,1H,CH),3.43-3.53(m,3H,CH₂and CH),3.57-3.59(m,2H,CH₂),3.63-3.72(m,10H,CH₂),4.20(d,J＝2.5Hz,2H,CH₂),5.34-5.35(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),13.5,19.3,20.8,22.2,23.6,24.1,25.9,26.3,29.4,30.0,31.1,31.4,31.6,36.3, 37.1,39.9,42.1,42.4,43.8,49.8,56.7,57.4,58.2,68.9,69.9,70.2,70.40,70.44,71.3,71.4,74.5, 75.1,79.5,121.3,140.7；HRMS calcd for C₃₆H₆₀O₆[M+Na]⁺:611.4282；found:611.4289。

TC568。¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H,CH₃),0.88-0.99(m,1H,CH₂),1.01(s,3H,CH₃),1.02-1.23(m,4H,CH₂and CH),1.26(s,3H,CH₃),1.28-1.76(m,20H,CH₂andCH),1.81-1.86(m,2H,CH₂),1.95-2.00(m,1H,CH₂),2.06-2.10(m,1H,CH₂),2.20-2.31(m,2H, CH₂),2.45(t,J＝2.5Hz,1H,CH),3.43-3.55(m,3H,CH₂and CH),3.57-3.60(m,2H,CH₂),3.63-3.72(m,14H,CH₂),4.21(d,J＝2.5Hz,2H,CH₂),5.34-5.35(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),13.5,19.3,20.8,22.2,23.7,24.1,26.0,26.3,29.5,30.0,31.2,31.5,31.7,36.4, 37.1,40.0,42.1,42.5,43.8,49.9,56.8,57.4,58.3,69.0,69.9,70.3,70.43,70.44,70.47,71.4,71.6, 74.5,75.1,79.5,121.5,140.7；HRMS calcd for C₃₈H₆₄O₇[M+Na]⁺:655.4544；found:655.4550。

TC569。¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H,CH₃),0.88-0.99(m,1H,CH₂),1.01(s,3H,CH₃),1.02-1.23(m,4H,CH₂and CH),1.26(s,3H,CH₃),1.28-1.76(m,20H,CH₂andCH),1.82-1.86(m,2H,CH₂),1.95-2.00(m,1H,CH₂),2.06-2.10(m,1H,CH₂),2.21-2.32(m,2H, CH₂),2.45(t,J＝2.5Hz,1H,CH),3.43-3.54(m,3H,CH₂and CH),3.57-3.74(m,20H,CH₂),4.21 (d,J＝2.5Hz,2H,CH₂),5.35-5.36(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),13.5,19.3, 20.8,22.2,23.7,24.2,26.0,26.3,29.5,30.0,31.2,31.5,31.7,36.4,37.1,40.0,42.1,42.5,43.8,49.9, 56.8,57.4,58.3,69.0,69.9,70.3,70.46,70.48,70.51,71.4,71.6,74.5,75.2,79.6,121.5,140.7； HRMS calcd for C₄₀H₆₈O₈[M+Na]⁺:699.4806；found:699.4800。

TC570。¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H,CH₃),0.88-0.99(m,1H,CH₂),1.01(s,3H,CH₃),1.02-1.23(m,4H,CH₂and CH),1.26(s,3H,CH₃),1.28-1.76(m,20H,CH₂andCH),1.82-1.86(m,2H,CH₂),1.95-2.02(m,1H,CH₂),2.07-2.10(m,1H,CH₂),2.21-2.32(m,2H, CH₂),2.45(t,J＝2.5Hz,1H,CH),3.43-3.55(m,3H,CH₂and CH),3.57-3.59(m,2H,CH₂),3.63-3.72(m,22H,CH₂),4.21(d,J＝2.5Hz,2H,CH₂),5.35-5.36(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),13.5,19.3,20.8,22.3,23.7,24.2,26.0,26.4,29.5,30.0,31.2,31.5,31.7,36.4, 37.2,40.0,42.2,42.6,43.9,49.9,56.8,57.4,58.3,69.0,69.9,70.3,70.49,70.50,70.53,71.4,71.6, 74.5,75.1,79.6,121.5,140.7；HRMS calcd for C₄₂H₇₂O₉[M+Na]⁺:744.5141；found:743.5070。

如上所示的反应方程式为TC819的合成路线，反应条件具体如下：

(a)5-amino-1-pentanol,NaBH(OAc)₃,AcOH,DCE,r.t.,7days；

(b)Boc₂O,DCM,r.t.,2h；

(c)TsCl,Et₃N,DMAP,DCM,r.t.,1h；

(d)ethylene glycol,NaH(60％),DMF,70℃,6h；

(e)3-bromo-1-propyne,NaH(60％),THF,50℃,2h；

(f)2N HCl in dioxane,r.t.,2h.

8.6g中间体1、8.2g 5-氨基-1-戊醇、4.8g冰醋酸溶于100mL1,2-二氯乙烷(DCE)中，在7天内分批加入17.0g三乙酰氧基硼氢化钠(NaBH(OAc)₃)，室温搅拌。反应结束后加入100mL饱和碳酸氢钠溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和碳酸氢钠水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝10:1) 分离得到纯净的中间体13。¹H NMR(500MHz,CDCl₃),(ppm)0.06(s,6H,CH₃),0.70(s,3H, CH₃),0.72(s,3H,CH₃),0.89(s,9H,CH₃),0.90-0.98(m,1H,CH₂),0.99(s,3H,CH₃),1.00(s,3H, CH₃),1.01-1.25(m,4H,CH₂and CH),1.05(d,J＝6.5Hz,3H,CH₃),1.22(d,J＝6.5Hz,3H,CH₃),1.28-1.73(m,21H,CH₂and CH),1.78-1.82(m,1H,CH₂),1.92-1.97(m,2H,CH₂),2.15-2.18(m, 1H,CH₂),2.24-2.29(m,1H,CH₂),2.51-2.67(m,1H,CH₂),2.72-2.84(m,1H,CH₂),3.45-3.51(m, 1H,CH),3.62(t,J＝8.5Hz,2H,CH₂),5.30-5.31(m,1H,CH)；¹³C NMR(125MHz,CDCl₃)δ (ppm)-4.6,12.0,12.4,17.6,18.2,19.4,20.8,21.0,23.1,23.4,24.2,25.9,26.1,26.7,27.4,28.8, 31.7,31.8,32.0,32.2,37.3,39.1,39.7,42.7,40.9,50.0,54.9,55.3,56.2,56.4,61.7,62.0,72.6, 120.9,141.6；HRMS calcd for C32H59NO2Si[M+H]+:518.4388；found:518.4390.

在2.07g中间体13的20mL DCM溶液中加入959mg二碳酸二叔丁酯(Boc₂O)，室温搅拌2小时。反应结束后浓缩，用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝5:1)分离得到纯净的中间体14。¹H NMR(500MHz,CDCl₃),(ppm)0.05(s,6H,CH₃),0.72 and 0.76(s,3H,CH₃),0.89(s,9H,CH₃),0.90-0.95(m,1H,CH₂),0.99 and 1.00(s,3H,CH₃),1.01-1.24 (m,4H,CH₂and CH),1.33-1.34(m,3H,CH),1.45(s,3H CH₃),1.50-1.98(m,21H,CH₂andCH), 2.15-2.18(m,1H,CH₂),2.24-2.29(m,1H,CH₂),3.45-3.47(m,1H,CH),3.65(t,J＝8.5Hz,2H, CH₂),5.30-5.31(m,1H,CH)；¹³C NMR(125MHz,CDCl₃)δ(ppm)-4.6,12.3,18.2,19.4,20.9, 21.0,23.7,23.9,24.2,25.9,28.5,28.6,29.2,30.3,31.8,32.0,32.4,36.5,37.3,39.1,41.4,41.9,42.0, 42.7,50.0,51.8,54.8,55.3,57.0,62.6,62.7,72.5,121.0,141.5,156.0；HRMS calcd for C₃₇H₆₇NO₄Si[M+Na]⁺:640.4732；found:640.4727.

1.23g中间体14、404mg三乙胺及24mg DMAP溶于10mL DCM中，加入570mg TsCl，室温搅拌1小时。反应结束后加入20mL饱和碳酸氢钠溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和碳酸氢钠水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝7:1)分离得到纯净的中间体15。¹H NMR(500MHz,CDCl₃), (ppm)0.05(s,6H,CH₃),0.71 and 0.75(s,3H,CH₃),0.89(s,9H,CH₃),0.99 and 1.00(s,3H, CH₃),1.08-1.29(m,10H,CH₂and CH),1.43(s,3H CH₃),1.50-1.98(m,17H,CH₂and CH), 2.15-2.29(m,2H,CH₂),2.45(s,3H,CH₃),2.71-2.88(m,1H,CH),2.97-3.11(m,1H,CH), 3.46-3.50(m,1H,CH),4.01-4.11(m,2H,CH₂),4.25-4.31(m,1H,CH),5.30-5.31(m,1H,CH), 7.34(d,J＝8.0Hz,2H,CH),7.79(d,J＝8.0Hz,2H,CH)；¹³C NMR(125MHz,CDCl₃)δ(ppm) -4.6,12.3,18.2,19.4,20.9,21.6,23.3,23.9,25.9,28.5,28.6,29.2,31.8,32.0,36.5,37.3,39.2,41.4, 41.9,42.7,50.1,51.8,54.8,70.4,72.5,121.0,127.8,129.8,133.1,141.5,144.7；HRMS calcd for C₄₄H₇₃NO₆SSi[M+Na]⁺:794.4820；found:794.4820.

在996mg乙二醇和10mL DMF的混合溶液中加入300mg NaH(60％)，室温搅拌15 分钟，然后加入1.16g中间体15，加热至70℃反应6小时。反应结束后加入20mL饱和氯化铵溶液淬灭反应，用二氯甲烷萃取三次，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝3:1)分离得到纯净的中间体16。¹H NMR(500MHz,CDCl₃), (ppm)0.06(s,6H,CH₃),0.72 and 0.76(s,3H,CH₃),0.89(s,9H,CH₃),0.99 and 1.00(s,3H, CH₃),1.02-1.35(m,10H,CH₂and CH),1.45(s,3H CH₃),1.52-1.98(m,15H,CH₂and CH), 2.15-2.27(m,2H,CH₂),3.02-3.24(m,1H,CH),3.46-3.55(m,5H,CH₂and CH),3.71-3.74(m,2H,CH₂),5.30-5.31(m,1H,CH)；¹³C NMR(125MHz,CDCl₃)δ(ppm)-4.6,12.1,12.2,18.1,19.30,19.34,19.8,20.2,20.9,23.8,24.0,25.8,28.46,28.51,28.6,29.1,29.3,30.2,31.70,31.77,31.9, 36.4,37.2,39.2,41.4,41.8,42.0,42.7,50.1,51.7,54.7,54.9,56.0,57.0,61.6,71.1,71.7,72.5,78.7, 78.9,120.1,141.4,155.1,155.9；HRMS calcd forC₃₉H₇₁NO₅Si[M+Na]⁺:684.4994；found: 684.4991.

在662mg中间体16的10mL THF溶液中加入80mg NaH(60％)，室温搅拌15分钟，然后加入357mg 3-溴丙炔，加热至50℃反应2小时。反应结束后加入20mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝7:1)分离得到纯净的中间体17。 HRMScalcd for C₄₂H₇₃NO₅Si[M+Na]⁺:722.5150；found:722.5146.

在140mg中间体17的2mL二氧六环溶液中加入2mL 4M HCl的二氧六环溶液，室温搅拌2小时。反应结束后浓缩、用柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝10:1) 分离得到纯净的中间体TC819。¹H NMR(600MHz,CDCl₃),(ppm)0.75 and 0.76(s,3H, CH₃),0.78-1.00(m,2H,CH),1.02 and 1.03(s,3H,CH₃),1.06-1.31(m,7H,CH₂),1.34 and 1.41(s, 3H,CH₃),1.43-1.68(m,15H,CH₂and CH),1.72-1.89(m,9H,CH₂and CH),1.93-2.02(m,3H, CH₂),2.20-2.30(m,2H,CH₂),2.58(t,J＝2.4Hz,1H,CH),2.89-2.96(m,2H,CH₂),3.21-3.24(m, 1H,CH),3.34-3.36(m,1H,CH₂),3.45-3.52(m,4H,CH₂),3.58-3.63(m,3H,CH₂),3.70-3.73(m, 3H,CH₂),4.21(d,J＝2.4Hz,2H,CH₂),5.34-5.35(m,1H,CH)；¹³C NMR(150MHz,CDCl₃), (ppm)11.4,11.9,14.8,15.6,18.85,18.87,20.3,20.4,23.0,23.5,23.6,23.8,25.26,25.29,25.9, 28.41,28.43,30.6,31.20,31.24,31.26,31.30,36.0,36.1,36.81,36.84,37.2,38.6,41.3,41.9,42.1, 42.2,43.0,49.4,49.6,51.0,52.0,53.5,55.4,55.9,56.6,57.9,60.8,68.6,69.4,70.4,70.70,70.72, 71.9,74.5,78.9,120.67,120.70,140.5；HRMS calcd for C₃₁H₅₁NO₃[M+H]⁺:486.3942；found: 486.3932.

如上所示的反应方程式为TC818的合成路线，反应条件具体如下：

(a)p-TsOH,MeOH,60℃,12h；

(b)TBSCl,imidazole,DMAP,DMF/THF,r.t.,1h；

(c)LiAlH₄,THF,50℃,1h；

(d)TsCl,Et₃N,DMAP,DCM,r.t.,1h；

(e)allylmagnesium bromide,Li₂CuCl₄,THF,-78℃ to r.t.,12h；

(f)i.9-BBN,THF,r.t.,1h；ii.1M NaOH,H₂O₂,r.t.,1h；

(g)ethylene glycol,NaH(60％),DMF,70℃,6h；

(h)3-bromo-1-propyne,NaH(60％),THF,50℃,2h；

(i)TBAF,THF,r.t.,1h.

在2.0g 3-羟基-5-胆烯酸的30mL甲醇溶液中加入95mg对甲苯磺酸(p-TsOH)，加热至60℃搅拌2小时。反应结束后浓缩除去溶剂，用100mL乙酸乙酯溶解，依次用饱和碳酸氢钠水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品溶于3mLDMF和5mLTHF的混合溶剂中，加入680mg咪唑、64mg DMAP及1.5g TBSCl，室温搅拌1小时。反应结束后加入20mL饱和碳酸氢钠溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和碳酸氢钠水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300 目硅胶，洗脱剂为正己烷：乙酸乙酯＝20:1)分离得到纯净的中间体18。¹H NMR(500MHz, CDCl₃),(ppm)0.06(s,6H,CH₃),0.67(s,3H,CH₃),0.89(s,9H,CH₃),0.90(d,J＝6.5Hz,3H, CH₃),0.93-0.97(m,1H,CH₂),1.00(s,3H,CH₃),1.01-1.60(m,H,CH₂and CH),1.68-1.73(m,1H, CH),1.78-1.86(m,2H,CH₂),1.93-2.06(m,4H,CH₂),2.14-2.18(m,1H,CH₂),2.24-2.30(m,1H, CH₂),3.44-3.51(m,1H,CH),4.91-5.01(m,2H,CH₂),5.30-5.32(m,1H,CH)；5.77-5.85(m,1H, CH),¹³C NMR(125MHz,CDCl₃)δ(ppm)-4.6,11.8,18.2,18.7,19.4,21.1,24.3,25.6,25.9,28.2, 29.4,31.9,31.9,32.1,33.9,35.7,35.8,36.6,37.4,39.8,42.3,42.8,50.2,56.1,56.8,72.6,114.1,121.1,139.2,141.5.

冰浴下在2.66g中间体18的30mLTHF溶液中缓慢加入570mg氢化锂铝(LiAlH₄)，加热至50℃反应1小时。反应结束后在冰浴下小心加入30mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析 (200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝10:1)分离得到纯净的中间体19。

2.51g中间体19、1.01g三乙胺及61mg DMAP溶于30mL DCM中，加入1.9g TsCl，室温搅拌1小时。反应结束后加入30mL饱和碳酸氢钠溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和碳酸氢钠水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝10:1)分离得到纯净的中间体20。¹H NMR(500MHz,CDCl₃), (ppm)0.06(s,6H,CH₃),0.64(s,3H,CH₃),0.87(d,J＝6.5Hz,3H,CH₃),0.89(s,9H,CH₃), 0.91-0.97(m,1H,CH),0.99(s,3H,CH₃),1.00-1.57(m,16H,CH₂and CH),1.66-1.82(m,4H, CH₂),1.93-1.99(m,2H,CH₂),2.14-2.18(m,1H,CH₂),2.23-2.29(m,1H,CH₂),2.45(s,3H,CH₃), 3.44-3.50(m,1H,CH),3.96-4.04(m,2H,CH₂),5.31-5.32(m,1H,CH),7.34(d,J＝8.0Hz,2H, CH),7.79(d,J＝8.0Hz,2H,CH)；¹³C NMR(125MHz,CDCl₃),(ppm)-4.6,11.8,18.3,18.5, 19.4,21.0,21.6,24.2,25.6,26.0,28.1,31.4,32.0,32.1,35.2,36.5,37.4,39.8,42.3,42.8,50.2,55.8, 56.8,71.2,72.6,121.1,127.9,129.8,133.3,141.6,144.6.

在干燥的、充满氮气的反应瓶中将2.63g中间体20溶于20mL无水THF中，在-78℃下向此溶液依次滴加1mL 0.1M四氯铜酸锂的THF溶液及4.9mL 1.7M烯丙基溴化镁的THF 溶液，滴加完毕后缓慢升至室温并继续搅拌12小时。反应结束后加入30mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝100:1)分离得到纯净的中间体21。¹HNMR(500MHz,CDCl₃),(ppm)0.06(s,6H,CH₃),0.67(s,3H,CH₃),0.89(s,9H,CH₃), 0.91(d,J＝6.5Hz,3H,CH₃),0.99(s,3H,CH₃),1.01-1.60(m,20H,CH₂and CH),1.69-1.73(m, 1H,CH₂),1.78-1.85(m,2H,CH₂),1.93-2.06(m,4H,CH₂),2.14-2.18(m,1H,CH₂),2.24-2.29(m,1H,CH₂),3.46-3.90(m,1H,CH),4.91-5.01(m,2H,CH₂),5.30-5.32(m,1H,CH),5.77-5.85(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),(ppm)-4.6,11.8,18.2,18.7,19.4,21.1,24.3,25.6,25.9, 28.2,29.4,31.91,31.94,32.1,33.9,35.7,35.8,36.6,37.4,39.8,42.3,42.8,50.2,56.1,56.8,72.6, 114.1,121.1,139.2,141.5.

在干燥的、充满氮气的反应瓶中将1.79g中间体21溶于25mL无水THF中，向此溶液滴加36mL 0.5M 9-BBN的THF溶液，室温搅拌1小时。之后将反应降温至0℃，缓慢滴加 4mL1M氢氧化钠溶液及4mL 30％过氧化氢，继续搅拌1小时。反应结束后加入60mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝10:1)分离得到纯净的中间体22。¹H NMR(500MHz,CDCl₃),(ppm)0.06(s,6H,CH₃),0.67(s,3H,CH₃),0.89(s, 9H,CH₃),0.91(d,J＝6.0Hz,3H,CH₃),1.00(s,3H,CH₃),1.03-1.58(m,26H,CH₂and CH), 1.68-1.73(m,1H,CH₂),1.78-1.84(m,2H,CH₂),1.94-2.02(m,2H,CH₂),2.14-2.18(m,1H,CH₂), 2.23-2.29(m,1H,CH₂),3.45-3.51(m,1H,CH),3.64(t,J＝3.5Hz,2H,CH₂),5.30-5.32(m,1H, CH)；¹³C NMR(125MHz,CDCl₃),(ppm)-4.6,11.8,18.3,18.7,19.4,21.1,24.3,25.8,25.94, 26.00,28.2,29.9,31.90,31.94,32.1,32.8,35.7,35.9,36.6,37.4,39.8,42.3,42.8,50.2,56.1,56.8, 63.1,72.6,121.1,141.6.

2.82g中间体22、1.01g三乙胺及61mg DMAP溶于30mL DCM中，加入1.9g TsCl，室温搅拌1小时。反应结束后加入30mL饱和碳酸氢钠溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和碳酸氢钠水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝10:1)分离得到纯净的中间体23。¹H NMR(500MHz,CDCl₃), (ppm)0.06(s,6H,CH₃),0.66(s,3H,CH₃),0.89(s,9H,CH₃),0.90(d,J＝6.0Hz,3H,CH₃), 0.92-0.97(m,2H,CH₂),1.00(s,3H,CH₃),1.04-1.84(m,28H,CH₂and CH),1.94-2.01(m,2H, CH₂),2.15-2.18(m,1H,CH₂),2.24-2.29(m,1H,CH₂),2.45(s,3H,CH₃),3.45-3.51(m,1H,CH), 4.01(t,J＝6.5Hz,2H,CH₂),5.31-5.32(m,1H,CH),7.34(d,J＝8.0Hz,2H,CH),7.79(d,J＝8.0 Hz,2H,CH)；¹³C NMR(125MHz,CDCl₃),(ppm)-4.6,11.8,18.2,18.6,19.4,21.0,21.6,24.2, 25.3,25.7,25.9,28.2,28.8,29.4,31.8,31.9,32.0,35.6,35.7,36.5,37.3,39.7,42.2,42.8,50.1,56.0, 56.7,70.6,72.6,121.1,127.8,129.7,133.1,141.4,144.5.

在542mg乙二醇和5mL DMF的混合溶液中加入146mg NaH(60％)，室温搅拌15 分钟，然后加入489mg中间体23，加热至70℃反应6小时。反应结束后加入30mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝5:1)分离得到纯净的中间体24。¹HNMR(500MHz,CDCl₃),(ppm)0.06(s,6H,CH₃),0.67(s,3H,CH₃),0.89(s,9H, CH₃),0.91(d,J＝6.0Hz,3H,CH₃),1.00(s,3H,CH₃),1.02-1.61(m,25H,CH₂and CH),1.70-1.73 (m,3H,CH₂),1.78-1.86(m,2H,CH₂),1.94-2.01(m,2H,CH₂),2.11-2.18(m,2H,CH₂),2.24-2.29 (m,1H,CH₂),3.45-3.51(m,3H,CH),3.54(t,J＝4.5Hz,2H,CH₂),3.73(br,2H,CH₂),5.31-5.32(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),(ppm)-4.6,11.8,18.2,18.7,19.4,21.0,24.3,25.92,25.97,26.1,28.2,29.7,30.0,31.89,31.93,32.1,35.7,35.9,36.6,37.4,39.8,42.3,42.8,50.2,56.1, 56.8,61.8,71.4,71.7,72.6,121.1,141.5.

在112mg中间体24的2mL THF溶液中加入16mg NaH(60％)，室温搅拌15分钟，然后加入24mg 3-溴丙炔，加热至50℃反应2小时。反应结束后加入10mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝20:1)分离得到纯净的中间体25。¹H NMR(500MHz,CDCl₃),(ppm)0.06(s,6H,CH₃),0.67(s,3H,CH₃),0.89(s,9H,CH₃), 0.90(d,J＝6.5Hz,3H,CH₃),0.93-0.98(m,1H,CH₂),1.00(s,3H,CH₃),1.01-1.60(m,24H,CH₂ and CH),1.69-1.73(m,1H,CH₂),1.78-1.86(m,2H,CH₂),1.94-2.02(m,2H,CH₂),2.14-2.18(m, 1H,CH₂),2.24-2.29(m,1H,CH₂),2.43(t,J＝2.5Hz,1H,CH),3.44-3.51(m,3H,CH₂and CH),3.60-3.62(m,2H,CH₂),3.68-3.71(m,2H,CH₂),4.21(d,J＝2.5Hz,2H,CH₂),5.31-5.32(m,1H, CH)；¹³C NMR(125MHz,CDCl₃),-4.6,11.8,18.3,18.7,19.4,21.0,24.3,25.9,26.1,28.2, 29.6,30.0,31.87,31.92,32.06,35.7,35.9,36.6,37.4,39.8,42.3,42.8,50.2,56.0,56.8,58.4,69.1, 69.8,71.6,72.6,74.4,79.7,121.1,141.5.

在120mg中间体25的2mL THF溶液中加入1M四丁基氟化铵(TBAF)的THF溶液0.6mL，室温搅拌1小时。反应结束后浓缩、用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝3:1)分离得到纯净的中间体TC818。¹H NMR(500MHz,CDCl₃),(ppm)0.67(s, 3H,CH₃),0.91(d,J＝6.0Hz,3H,CH₃),0.92-0.99(m,1H,CH₂),1.00(s,3H,CH₃),1.02-1.62(m, 24H,CH₂and CH),1.78-1.86(m,5H,CH₂),1.94-2.02(m,2H,CH₂),2.20-2.31(m,2H,CH₂),2.43(t,J＝2.5Hz,1H,CH),3.46(t,J＝2.0Hz,2H,CH₂),3.48-3.55(m,1H,CH),3.60-3.62(m,2H,CH₂),3.68-3.70(m,2H,CH₂),4.21(d,J＝2.0Hz,2H,CH₂),5.34-5.35(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),(ppm)11.8,18.7,19.4,21.0,24.2,25.9,26.1,28.2,29.6,29.9,31.6,31.8, 35.7,35.8,36.4,37.2,39.7,42.23,42.26,50.1,56.0,56.7,58.4,69.1,69.8,71.6,71.7,74.4,79.6, 121.6,140.7.

如上所示的反应方程式为TC667的合成路线，反应条件具体如下：

(a)NaH(60％),THF,reflux,6h；

(b)HATU,DIPEA,DCM,r.t.,2h.

在144mg 1-炔丙基二乙二醇的5mL THF溶液中加入120mg NaH(60％)，室温搅拌15分钟，然后加入152mg溴乙酸，加热回流6小时。反应结束后加入10mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝50:1)分离得到纯净的中间体26。¹H NMR(500MHz,CDCl₃),δ(ppm)2.46(t,J＝2.0Hz,1H,CH),3.70–3.78(m,8H,CH₂),4.19(s, 2H,CH₂),4.21(d,J＝2.5Hz,CH₂)；¹³C NMR(125MHz,CDCl₃),δ(ppm)58.5,68.6,69.0,70.4,70.5,71.2,74.8,79.5,173.6；HRMS calcd for C₉H₁₄O₅[M+Na]⁺:225.0733；found:225.0746.

73mg中间体26及153mg O-(7-氮苯并三氮唑)-N,N,N,N-四甲基脲六氟磷酸酯(HATU) 溶于3mL DCM中，室温搅拌0.5小时。向此溶液加入123mg环巴胺和58mg二异丙基乙胺(DIPEA)的2mL DCM溶液，室温搅拌2小时。反应结束后加入10mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝20:1)分离得到纯净的中间体TC667。

如上所示的反应方程式为TC558的合成路线，反应条件具体如下：

(a)p-TsOH,MeOH,60℃,12h；

(b)TBSCl,imidazole,DMAP,DMF/THF,r.t.,1h；

(c)LiAlH₄,THF,50℃,1h；

(d)TsCl,Et₃N,DMAP,DCM,r.t.,1h；

(e)1-propargyl diethylene glycol,NaH(60％),DMF,70℃,4-6h；

(f)TBAF,THF,r.t.,1h.

在1.88g石胆酸的30mL甲醇溶液中加入95mg对甲苯磺酸(p-TsOH)，加热至60℃搅拌2小时。反应结束后浓缩除去溶剂，用100mL乙酸乙酯溶解，依次用饱和碳酸氢钠水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品溶于3 mL DMF和5mL THF的混合溶剂中，加入680mg咪唑、64mg DMAP及1.5g TBSCl，室温搅拌1小时。反应结束后加入20mL饱和碳酸氢钠溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和碳酸氢钠水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝100:1)分离得到纯净的中间体27。

冰浴下在2.52g中间体27的30mL THF溶液中缓慢加入570mg氢化锂铝(LiAlH₄)，加热至50℃反应1小时。反应结束后在冰浴下小心加入30mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析 (200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝20:1)分离得到纯净的中间体28。

1.43g中间体28、1.01g三乙胺及37mg DMAP溶于30mL DCM中，加入1.14g TsCl，室温搅拌1小时。反应结束后加入30mL饱和碳酸氢钠溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和碳酸氢钠水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝20:1)分离得到纯净的中间体29。¹H NMR(500MHz,CDCl₃), (ppm)0.06(s,6H,CH₃),0.59(s,3H,CH₃),0.84(d,J＝6.5Hz,3H,CH₃),0.89(s,12H,CH₃), 0.92-1.55(m,24H,CH₂and CH),1.63-1.85(m,5H,CH₂),1.89-1.93(m,1H,CH₂),2.45(s,3H, CH₃),3.55-3.61(m,1H,CH₂),3.96-4.04(m,2H,CH₂),7.34(d,J＝8.0Hz,2H,CH),7.79(d,J＝8.0Hz,2H,CH)；¹³C NMR(125MHz,CDCl₃),(ppm)-4.6,12.0,18.3,18.4,20.8,21.6,23.4,24.2,25.5,26.0,26.4,27.3,28.2,31.0,31.4,34.5,35.2,35.5,35.8,36.9,40.1,40.2,42.2,42.6,56.0, 56.3,71.2,72.8,127.8,129.7,133.3,144.5.

在58mg 1-炔丙基二乙二醇的5mL THF溶液中加入24mg NaH(60％)，室温搅拌15分钟，然后加入126mg中间体29，加热回流6小时。反应结束后加入10mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝3:1)分离得到纯净的中间体30。¹H NMR(500MHz,CDCl₃),(ppm)0.06(s,6H,CH₃),0.63(s,3H,CH₃),0.89-0.91(m,15H,CH₃), 0.92-1.68(m,27H,CH₂and CH),1.73-1.85(m,4H,CH₂),1.92-1.96(m,1H,CH₂),2.43(t,J＝2.5Hz,1H,CH),3.40-3.44(m,2H,CH₂),3.54-3.60(m,3H,CH₂and CH),3.64-3.66(m,2H,CH₂),3.68-3.72(m,4H,CH₂),4.21(d,J＝2.5Hz,2H,CH₂)；¹³C NMR(125MHz,CDCl₃),(ppm)-4.6,12.0,18.3,18.6,20.8,23.4,24.2,25.9,26.1,26.4,27.3,28.2,31.0,32.0,34.5,35.54,35.55,35.8, 36.9,40.11,40.17,42.2,42.6,56.1,56.4,58.4,69.1,70.0,70.4,70.6,72.0,72.8,74.4,79.6.

在60mg中间体30的2mL THF溶液中加入1M四丁基氟化铵(TBAF)的THF溶液0.6 mL，室温搅拌1小时。反应结束后浓缩、用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝1:1)分离得到纯净的中间体TC558。¹H NMR(500MHz,CDCl₃),(ppm)0.64(s,3H, CH₃),0.90-0.91(m,6H,CH₃),0.96-1.56(m,22H,CH₂and CH),1.64-1.66(m,2H,CH₂),1.72-1.84 (m,4H,CH₂),1.95-1.97(m,1H,CH₂),2.44(t,J＝2.5Hz,1H,CH),3.39-3.44(m,2H,CH₂), 3.59-3.65(m,5H,CH₂and CH),3.69-3.70(m,4H,CH₂),4.20-4.21(m,2H,CH₂)；¹³C NMR(125 MHz,CDCl₃),(ppm)11.9,18.5,20.7,23.3,24.1,26.0,26.3,27.1,28.2,30.4,31.9,34.5,35.3,35.5,35.7,36.3,40.1,40.3,42.0,42.6,56.0,56.4,58.3,69.0,70.0,70.3,70.5,71.7,71.9,74.5,79.5； HRMS calcd for C₃₁H₅₂O₄[M+Na]⁺:511.3758；found:511.3677.

如上所示的反应方程式为TC828的合成路线，反应条件具体如下：

(a)Et₃SiCl,imidazole,THF/DMF,r.t.,1h；

(b)I₂,PPh₃,imidazole,THF/DCM,0℃ to r.t.,1h；

(d)i.n-BuLi,HMPA,THF,-78℃ to 0℃,1h；ii.TBAF,THF,r.t.,1h；

(e)3-bromo-1-propyne,NaH(60％),THF,50℃,2h；

(f)i.n-BuLi,THF,-78℃,1h；ii.TMSCl,-78℃ to 0℃,1h；

(g)p-TsOH,MeOH,r.t.,1h；

(h)CBr₄,PPh₃,DCM,-10℃ to r.t.,6h；

(i)magnesium,THF,reflux,3h；

(j)TBAF,THF,r.t.,2h.

8.48g二乙二醇和2.0g咪唑溶于20mL DMF和30mL THF的混合溶剂中，冰浴下向此溶液缓慢滴加2.92g三乙基氯硅烷(Et₃SiCl)的20mL THF溶液，滴加完毕后室温搅拌1小时。反应结束后加入100mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝3:1)分离得到纯净的中间体31。

4.4g中间体31、1.7g咪唑及5.5g三苯基膦(PPh₃)溶于100mL DCM中，冰浴下向此溶液缓慢滴加5.08g单质碘(I₂)的30mL THF溶液，滴加完毕后室温搅拌1小时。反应结束后加入100mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝10:1)分离得到纯净的中间体32。

0℃下在3.5g 3-丁炔-1-醇的100mL DCM溶液中加入1.0g p-TsOH和4.2g 3,4-二氢-2H- 吡喃(DHP)，室温搅拌1小时。反应结束后加入100mL饱和碳酸氢钠溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和碳酸氢钠水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝20:1)分离得到纯净的中间体33。

在干燥的、充满氮气的反应瓶中将3.08g中间体33溶于100mL无水THF中，在-78℃下向此溶液缓慢滴加2.4M正丁基锂(n-BuLi)8.3mL并搅拌0.5小时。在此温度下依次加入3.58g六甲基磷酰三胺(HMPA)、4.95g 32，升温至0℃搅拌1小时。反应结束后加入 100mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩，用100mLTHF重新溶解，加入1M TBAF的THF溶液30mL，室温搅拌1小时。反应结束后浓缩，粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝3:1)分离得到纯净的中间体34。HRMScalcd for C₁₃H₂₂O₄[M+Na]⁺:265.1410；found:265.1413.

在1.37g中间体34的40mL THF溶液中加入292mg NaH(60％)，室温搅拌15分钟，然后加入2.38g 3-溴丙炔，加热至50℃反应2小时。反应结束后加入100mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝8:1)分离得到纯净的中间体35。 HRMScalcd for C₁₆H₂₄O₄[M+Na]⁺:303.1567；found:303.1597.

在干燥的、充满氮气的反应瓶中将1.24g中间体35溶于20mL无水THF中，在-78℃下向此溶液缓慢滴加1.6M正丁基锂(n-BuLi)3.0mL并搅拌0.5小时。在此温度下加入0.53 g三甲基氯硅烷(TMSCl)，缓慢升温至0℃继续搅拌1小时。反应结束后加入50mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝8:1)分离得到纯净的中间体36。

在915mg中间体36的30mL甲醇溶液中加入57mg p-TsOH，室温搅拌1小时。反应结束后浓缩除去溶剂，用二氯甲烷重新溶解，依次用饱和碳酸氢钠水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝5:1)分离得到纯净的中间体37。HRMS calcd for C₁₄H₂₄O₃[M+Na]⁺:291.1387；found:291.1398.

402mg中间体37及996mg四溴化碳(CBr₄)溶于15mL DCM中，在-10℃下向此溶液缓慢滴加786mg PPh₃的10mL DCM溶液，滴加完毕后缓慢升至室温搅拌6小时。反应结束后加入30mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝10:1)分离得到纯净的中间体38。¹H NMR(500MHz,CDCl₃),(ppm)0.18(s,9H,CH₃), 2.44-2.48(m,2H,CH₂),2.70-2.73(m,2H,CH₂),3.41(t,J＝7.5Hz,2H,CH₂),3.59(t,J＝7.5Hz, 2H,CH₂),3.65-3.69(m,4H,CH₂),4.21(s,2H,CH₂)；¹³C NMR(125MHz,CDCl₃),(ppm) -0.2,20.0,23.3,30.0,59.2,68.9,69.6,70.1,78.1,79.0,91.4,101.3；HRMS calcd for C₁₄H₂₃BrO₂Si [M+Na]⁺:353.0543；found:353.0542.

在干燥的、充满氮气的反应瓶中加入192mg镁屑。将1.32g中间体38溶于5mL无水THF中，该溶液的约1/4加至镁屑中，加入1滴1,2-二溴乙烷并适当加热引发格氏试剂的形成。引发成功后将剩余的溴代物溶液滴加至镁屑中，控制滴加速率以使反应液保持微沸。滴加完毕后继续加热回流2小时。将反应液冷却待用。

在干燥的、充满氮气的反应瓶中将430mg中间体1溶于5mL无水THF中，向此溶液滴加上述格氏试剂的THF溶液5mL，室温搅拌3小时。反应结束后加入30mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝7:1)分离得到纯净的中间体39。¹H NMR(500MHz,CDCl₃),(ppm)0.12(s,15H,CH₃),0.79(s,3H,CH₃),0.83(s,9H,CH₃), 0.86-0.91(m,1H,CH₂),0.94(s,3H,CH₃),0.98-1.18(m,4H,CH₂and CH),1.21(s,3H,CH₃), 1.29-1.83(m,21H,CH₂and CH),1.89-1.93(m,1H,CH₂),2.02-2.23(m,9H,CH₂),2.38-2.40(m, 5H,CH₂),3.38-3.45(m,1H,CH),3.49-3.53(m,2H,CH₂),3.59-3.63(m,2H,CH₂),4.15(s,2H,CH₂),5.25-5.26(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),(ppm)-4.6,0.2,13.45,13.52,18.16,18.22,19.3,20.0,20.8,22.4,23.7,25.5,25.8,27.9,31.2,31.7,32.0,36.5,37.3,40.1,41.8, 42.6,42.7,50.0,56.8,58.6,59.1,68.9,69.8,69.9,72.5,74.7,76.7,76.9,80.9,81.4,91.3,101.3, 120.9,141.4.

在68mg中间体39的2mL THF溶液中加入1M四丁基氟化铵(TBAF)的THF溶液0.6 mL，室温搅拌1小时。反应结束后浓缩、用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝1:1)分离得到纯净的中间体TC828。¹H NMR(600MHz,CDCl₃),(ppm)0.85(s,3H, CH₃),0.87-0.99(m,2H,CH₂),1.01(s,3H,CH₃),1.04-1.23(m,3H,CH₂and CH),1.27(s,3H, CH₃),1.40-1.85(m,17H,CH₂and CH),1.96-1.99(m,1H,CH₂),2.08-2.10(m,1H,CH₂),2.14-2.31 (m,4H,CH₂),2.43-2.45(m,3H,CH₂),3.49-3.58(m,3H,CH₂and CH),3.65-3.66(m,2H,CH₂), 3.69-3.71(m,2H,CH₂),4.22(d,J＝1.2Hz,2H,CH₂),5.35-5.36(m,1H,CH)；¹³C NMR(150 MHz,CDCl₃),(ppm)13.5,13.6,19.3,20.0,20.9,22.4,23.7,25.5,31.2,32.5,31.7,36.4,37.2,40.1,41.8,42.2,42.7,49.9,56.8,58.4,58.6,69.0,69.9,70.0,71.7,74.5,74.8,76.9,79.6,81.5,91.3, 121.5,140.7；HRMS calcd for C₃₂H₄₈O₄[M+Na]⁺:519.3445；found:519.3450.

如上所示的反应方程式为TC599及其同系物的合成路线，反应条件具体如下：

(a)Pd(PPh₃)₄,Na₂CO₃,toluene/H₂O,90℃,5h；

(b)Boc₂O,DCM,r.t.,2h；

(c)LiAlH₄,THF,reflux,2h；

(d)i,PhCHO,toluene,reflux,6h；ii,Boc₂O,r.t.,1h；iii,1M KHSO₄,r.t.,1h；

(e)NaBH₄,MeOH,40℃,1h；

(f)HATU,DIPEA,DCM,40℃,12h；

(g)oligo ethylene glycol,NaH(60％),THF,60℃,4-6h；

(h)DPPA,DBU,THF,r.t.to reflux,18-24h；

(i)2N HCl in dioxane,r.t.,2h.

在干燥的、充满氮气的反应瓶中加入2-氟-4-溴吡啶4.4g、3-甲酰基苯硼酸3.94g、四 (三苯基膦)钯577mg、碳酸氢钠6.1g、甲苯50mL和水100mL，加热至90℃搅拌5小时。反应结束后加入50mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝20:1)分离得到纯净的中间体40。¹H NMR(500MHz,CDCl₃),δ(ppm)7.19(s,1H,CH), 7.46-7.47(m,1H,CH),7.71(t,J＝7.5Hz,1H,CH),7.90-7.92(m,1H,CH),7.99-8.01(m,1H,CH), 8.15-8.16(m,1H,CH),8.33(d,J＝5.0Hz,1H,CH),10.12(s,1H,CH)；¹³C NMR(125MHz, CDCl₃),δ(ppm)107.3(d,J＝38.0Hz),119.4(d,J＝4.1Hz),127.7,130.0,131.0,132.7,137.1,138.0(d,J＝3.4Hz),148.3(d,J＝15.4Hz),152.5(d,J＝8.2Hz),164.4(d,J＝237.4Hz),191.5.

在22.8g反式1,4-环己二胺的200mL DCM溶液中加入10.8二碳酸二叔丁酯(Boc₂O)，室温搅拌2小时。反应结束后浓缩、用柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝7:1)分离得到纯净的中间体41。

在2.1g中间体41的50mL THF溶液中加入1.8g LiAlH₄，加热回流2小时。反应结束后在冰浴下小心加入50mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝3:1)分离得到纯净的中间体42。

12.8g中间体42及10.6g苯甲醛溶于50mL甲苯中加热回流6小时，使用分水器分出反应形成的水。之后降至室温并取下分水器，加入22g二碳酸二叔丁酯(Boc₂O)，室温搅拌1小时。然后加入50mL 1M硫酸氢钾水溶液，室温搅拌1小时。反应结束后分液，水相用乙酸乙酯洗涤2次后加入6M氢氧化钠水溶液调节pH大于12，用乙酸乙酯萃取3次，萃取的有机相用饱和氯化钠溶液洗涤，用无水硫酸钠干燥，过滤、浓缩得到粗产物。粗产品用柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝10:1)分离得到纯净的中间体43。

2.9g 40及3.0g中间体43溶于50mL甲醇中，加热至40℃搅拌0.5小时。向此溶液分批加入760mg硼氢化钠(NaBH₄)，继续在40℃搅拌0.5小时。反应结束后加入150mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝50:1)分离得到纯净的中间体44。¹H NMR(500MHz,CDCl₃),δ(ppm)1.24-1.51(m,4H,CH₂),1.46(s,3H,CH₃), 1.70-1.74(m,3H,CH₂and CH),2.04-2.07(m,2H,CH₂),2.44-2.49(m,1H,CH),2.70(s,3H,CH₃), 3.89(s,2H,CH₂),7.14(br,1H,CH),7.40-7.47(m,3H,CH),7.50-7.53(m,1H,CH),7.60(m,1H, CH),8.24-8.26(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)28.4,32.4,51.0,55.7,79.2, 107.0(d,J＝37.5Hz),119.5(d,J＝3.9Hz),125.5,126.6,129.2,129.3,137.1,141.8,147.8(d,J＝ 15.4Hz),153.9(d,J＝8.1Hz),155.5,164.4(d,J＝236.7Hz).HRMS calcd forC₂₄H₃₂FN₃O₂ [M+H]⁺:414.2551；found:414.2557.

2.55g 3-氯苯并[b]噻吩-2-甲酸及5.7g O-(7-氮苯并三氮唑)-N,N,N,N-四甲基脲六氟磷酸酯 (HATU)溶于25mL DCM中，室温搅拌0.5小时。向此溶液加入4.13g中间体44和3.22g 二异丙基乙胺(DIPEA)的25mL DCM溶液，加热至40℃搅拌过夜。反应结束后加入50mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝3:1)分离得到纯净的中间体45。¹H NMR(500MHz,CDCl₃),δ(ppm)1.24-1.51(m,4H,CH₂),1.46(s,3H,CH₃), 1.70-1.74(m,3H,CH₂and CH),2.04-2.07(m,2H,CH₂),2.44-2.49(m,1H,CH),2.70(s,3H,CH₃),3.89(s,2H,CH₂),7.14(br,1H,CH),7.40-7.47(m,3H,CH),7.50-7.53(m,1H,CH),7.60(m,1H, CH),8.24-8.26(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)28.4,32.4,51.0,55.7,79.2, 107.0(d,J＝37.5Hz),119.5(d,J＝3.9Hz),125.5,126.6,129.2,129.3,137.1,141.8,147.8(d,J＝ 15.4Hz),153.9(d,J＝8.1Hz),155.5,164.4(d,J＝236.7Hz)；HRMScalcd for C₃₃H₃₅ClFN₃O₃S [M+Na]⁺:630.1964；found:630.1955.

合成TC599及同系物的通用步骤：

在186mg乙二醇的10mL THF溶液中加入60mg NaH(60％)，室温搅拌15分钟，然后加入中间体45，加热至60℃反应6小时。反应结束后加入20mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析 (200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝1:1)分离得到纯净的中间体46。¹H NMR (500MHz,CDCl₃),δ(ppm)1.32-1.44(m,11H,CH₃and CH₂),1.63-1.72(m,4H,CH₂),1.84-1.96 (m,2H,CH₂),2.57 and 2.70(s,3H,CH₃),3.75-3.98(m,3H,CH₂and CH),4.49-4.50(m,2H,CH₂),4.66and 4.84(s,2H,CH₂),6.85-7.24(m,2H,CH),7.30-7.52(m,5H,CH),7.67-7.89(m,3H,CH), 8.12-8.16(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)13.9,20.7,28.1,30.5,38.2,44.8, 52.4,58.5,60.1,61.7,68.4,79.2,108.6,115.4,118.7,122.2,122.5,125.17,125.23,125.33,126.2, 127.3,129.0,129.7,135.4,136.9,138.0,138.8,146.5,151.1,155.1,163.3,164.2,170.8；HRMS calcd for C₃₅H₄₀ClN₃O₅S[M+H]⁺:650.2450；found:650.2439.

中间体47。¹H NMR(600MHz,CDCl₃),δ(ppm)1.25-1.43(m,10H,CH₂),1.63-1.73(m,4H, CH₂),1.85-1.96(m,2H,CH₂),2.57and 2.70(s,3H,CH₃),3.68-3.70(m,2H,CH₂),3.77(t,J＝4.8 Hz,2H,CH₂),3.89-3.90(m,2H,CH₂),4.54-4.55(m,2H,CH₂),4.67and 4.84(s,2H,CH₂), 6.87-7.23(m,2H,CH),7.32-7.55(m,5H,CH),7.69-7.71(m,1H,CH),7.82-7.92(m,2H,CH), 8.15-8.19(m,1H,CH)；¹³C NMR(150MHz,CDCl₃),δ(ppm)28.3,30.7,45.0,52.6,58.7,61.6, 65.1,69.6,72.4,79.4,108.7,115.4,115.6,119.0,122.5,122.7,125.4,125.6,126.4,127.2,127.5, 129.1,129.9,135.6,137.2,138.2,138.9,147.0,151.1,155.4,163.5,164.1；HRMS calcd for C₃₇H₄₄ClN₃O₆S[M+H]⁺:694.2712；found:694.2712.

中间体48。¹H NMR(600MHz,CDCl₃),δ(ppm)1.26-1.44(m,10H,CH₂),1.64-1.73(m,4H, CH₂),1.85-1.96(m,2H,CH₂),2.57and 2.71(s,3H,CH₃),3.61(t,J＝4.8Hz,2H,CH₂),3.69-3.79 (m,6H,CH₂),3.89-3.91(m,2H,CH₂),4.54-4.55(m,2H,CH₂),4.67and 4.84(s,2H,CH₂), 6.87-7.22(m,2H,CH),7.32-7.55(m,5H,CH),7.69-7.71(m,1H,CH),7.82-7.92(m,2H,CH), 8.14-8.18(m,1H,CH)；¹³C NMR(150MHz,CDCl₃),δ(ppm)28.3,29.4,30.8,45.0,52.6,58.7, 61.6,65.1,69.6,70.2,70.6,72.5,79.4,108.7,115.3,115.5,119.0,122.5,122.7,125.4,125.6,126.4, 127.2,127.4,129.1,129.9,135.6,137.2,138.3,138.9,146.9,151.0,155.4,163.5,164.1；HRMS calcd for C₃₉H₄₈ClN₃O₇S[M+H]⁺:738.2974；found:738.2976.

中间体49。¹H NMR(500MHz,CDCl₃),δ(ppm)1.25-1.53(m,4H,CH₂),1.46(s,9H,CH₃), 1.53-1.96(m,6H,CH₂and CH),2.70(s,3H,CH₃),3.60-3.76(m,16H,CH₂),3.91(s,2H,CH₂), 6.89-6.95(m,1H,CH),7.11(br,1H,CH),7.46-7.53(m,3H,CH),7.70(br,1H,CH),7.82-7.92(m, 2H,CH),8.16-8.22(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)28.4,61.7,69.6,70.3,70.6, 72.5,99.9,119.0,125.5,125.7,126.5,129.3,137.2,163.6；HRMScalcd for C₄₁H₅₂ClN₃O₈S [M+H]⁺:782.3236；found:782.3226.

325mg中间体46、550mg叠氮磷酸二苯酯(DPPA)及228mg 1,8-二氮杂二环十一碳 -7-烯(DBU)溶于5mL THF中，室温搅拌6-12小时，通过TLC检测46完全消失后，继续加热回流12小时。反应结束后加入20mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝3:1)分离得到纯净的中间体50。¹H NMR(500MHz,CDCl₃),δ(ppm) 1.26-1.43(m,4H,CH₂),1.40(s,9H,CH₃),1.64-1.96(m,6H,CH₂and CH),2.70(s,3H,CH₃), 3.61-3.78(m,2H,CH₂),3.90-3.92(m,2H,CH₂),4.15-4.20(m,2H,CH₂),6.89-6.96(m,1H,CH), 7.13(br,1H,CH),7.47-7.54(m,3H,CH),7.70(br,1H,CH),7.82-7.93(m,2H,CH),8.12-8.24(m, 1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)14.0,20.8,28.1,29.2,30.6,44.8,49.0,49.3,53.3,58.5,60.1,79.1,105.4,116.8,118.8,122.3,122.5,124.9,125.3,126.3,127.8,129.0,129.7,135.4, 137.0,137.3,137.9,138.8,151.9,155.2,162.3,163.4,170.8；HRMS calcdfor C₃₅H₃₉ClN₆O₄S [M+H]⁺:675.2515；found:675.2512.

中间体51。¹H NMR(500MHz,CDCl₃),δ(ppm)1.26-1.30(m,4H,CH₂),1.41(s,9H,CH₃), 1.66-1.96(m,6H,CH₂and CH),2.71(s,3H,CH₃),3.38(t,J＝5.0Hz,2H,CH₂),3.67-3.77(m,4H, CH₂),3.91(t,J＝5.0Hz,2H,CH₂),4.53-4.55(m,2H,CH₂),4.83(s,3H,CH₃),6.86-7.14(m,2H, CH),7.44-7.51(m,4H,CH),7.69(br,1H,CH),7.82-7.91(m,2H,CH),8.09-8.18(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)28.2,30.6,44.9,50.5,53.3,58.6,60.6,65.0,65.9,69.5,69.8, 79.3,98.5,108.6,115.4,119.9,120.0,125.2,125.4,125.9,126.3,129.0,129.7,129.9,137.0,146.8, 149.5,149.6,150.2,164.0；HRMS calcdfor C₃₇H₄₃ClN₆O₅S[M+H]⁺:719.2777；found:719.2772.

中间体52。¹H NMR(500MHz,CDCl₃),δ(ppm)1.26-1.30(m,4H,CH₂),1.41(s,9H,CH₃), 1.66-1.96(m,6H,CH₂and CH),2.71(s,3H,CH₃),3.38(t,J＝5.0Hz,2H,CH₂),3.67-3.77(m,8H, CH₂),3.91(t,J＝5.0Hz,2H,CH₂),4.53-4.55(m,2H,CH₂),4.83(s,3H,CH₃),6.86-7.14(m,2H, CH),7.44-7.51(m,4H,CH),7.69(br,1H,CH),7.82-7.91(m,2H,CH),8.09-8.18(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)28.3,30.7,45.0,50.5,58.6,65.1,69.7,69.9,70.6,79.3,108.7, 115.4,118.9,122.7,125.3,126.4,129.1,137.1,138.4,146.9,155.3,164.2；HRMS calcd for C₃₉H₄₇ClN₆O₆S[M+H]⁺:763.3039；found:763.3039.

中间体53。¹H NMR(500MHz,CDCl₃),δ(ppm)1.26-1.43(m,4H,CH₂),1.40(s,9H,CH₃), 1.64-1.96(m,6H,CH₂and CH),2.70(s,3H,CH₃),3.61-3.78(m,16H,CH₂),3.90-3.92(m,2H, CH₂),4.15-4.20(m,2H,CH₂),6.89-6.96(m,1H,CH),7.13(br,1H,CH),7.47-7.54(m,3H,CH), 7.70(br,1H,CH),7.82-7.93(m,2H,CH),8.12-8.24(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ (ppm)28.3,50.5,61.0,61.6,63.6,66.0,67.0,69.9,70.5,70.6,72.2,72.6,98.7,119.0,125.5,126.5, 129.3；HRMS calcd for C₄₁H₅₁ClN₆O₇S[M+H]⁺:807.3301；found:807.3310.

在134mg中间体50的2mL二氧六环溶液中加入2mL 4M HCl的二氧六环溶液，室温搅拌2小时。反应结束后浓缩、柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝30:1) 分离得到纯净的中间体TC599。¹H NMR(600MHz,CD₃OD),δ(ppm)1.07-1.13(m,2H,CH₂), 1.68-1.81(m,4H,CH₂),1.94-2.07(m,2H,CH₂),2.42 and 2.52(s,3H,CH₃),2.81-2.90(m,1H,CH),31.8-3.19(m,1H,CH),3.41-3.43(m,1H,CH),3.54-3.58(m,2H,CH₂),3.65-3.72(m,1H,CH),4.03-4.05(m,2H,CH₂),4.20 and 4.55(s,2H,CH₂),6.31-6.66(m,2H,CH),7.15-7.49(m,6H, CH),7.57-7.85(m,3H,CH)；¹³C NMR(150MHz,CD₃OD),δ(ppm)28.9,30.4,31.0,46.4,50.4, 50.8,57.5,60.1,62.4,64.5,73.5,107.9,116.9,120.3,123.6,124.5,126.6,126.9,127.4,128.4, 129.8,130.8,131.1,136.8,138.68,138.74,140.8,154.6,164.9,165.7；HRMS calcd for C₃₀H₃₁ClN₆O₂S[M+H]⁺:575.1990；found:575.1993.

TC600。¹H NMR(600MHz,CD₃OD),δ(ppm)1.18-1.31(m,2H,CH₂),1.80-2.21(m,6H,CH₂),2.54 and 2.65(s,3H,CH₃),2.90-2.97(m,1H,CH),3.32-3.34(m,1H,CH),3.39-3.40(m,2H, CH₂),3.76(t,J＝4.8 Hz,2H,CH₂),3.89-3.91(m,2H,CH₂),4.50 and 4.72(s,2H,CH₂),6.82-7.18 (m,3H,CH),7.24-7.32(m,2H,CH),7.49-7.63(m,4H,CH),7.79-7.82(m,1H,CH),7.91-8.01(m, 2H,CH),8.10-8.19(m,1H,CH)；¹³C NMR(150MHz,CD₃OD),δ(ppm)27.4,28.8,29.4,44.8, 50.4,55.9,58.6,60.8,65.3,69.2,69.9,72.1,108.1,115.2,118.7,119.9,120.0,122.0,122.9,123.2, 125.1,125.4,125.8,126.8,127.5,128.9,129.1,129.4,135.2,137.2,138.2,139.0,146.9,151.4, 152.71,152.75,164.2,164.5；HRMS calcd for C₃₂H₃₅ClN₆O₃S[M+H]⁺:619.2253；found: 619.2251.

TC801。¹H NMR(500MHz,CDCl₃ and CD₃OD),δ(ppm)1.24-1.34(m,2H,CH₂), 1.81-2.23(m,6H,CH₂),2.56 and 2.66(s,3H,CH₃),2.97-3.00(m,1H,CH),3.32-3.37(m,2H,CH₂),3.63-3.92(m,10H,CH₂),4.49 and 4.73(s,2H,CH₂),6.85-7.24(m,2H,CH),7.34-7.59(m, 5H,CH),7.76-7.97(m,3H,CH),8.11-8.17(m,1H,CH)；¹³C NMR(125MHz,CDCl₃ andCD₃OD), δ(ppm)26.7,28.2,28.8,49.7,58.0,62.2,64.6,68.8,69.1,69.6,69.7,107.6,109.2,114.6,121.5, 122.2,125.0,126.1,126.8,128.4,136.4,137.4,146.0；HRMS calcdfor C₃₄H₃₉ClN₆O₄S[M+H]⁺: 663.2515；found:663.2509.

TC588。¹H NMR(600MHz,CDCl₃),δ(ppm)1.22-1.28(m,2H,CH₂),1.85-2.23(m,6H,CH₂ and CH),2.58 and 2.68(s,3H,CH₃),3.32-3.34(m,2H,CH₂),3.61-3.89(m,16H,CH₂),4.12-4.25(m,2H,CH₂),4.46-4.49(m,2H,CH₂),6.81-6.99(m,1H,CH),7.10(br,1H,CH),7.27-7.62(m,3H,CH),7.81-7.91(m,2H,CH),8.00-8.12(m,1H,CH)；¹³C NMR(150MHz,CDCl₃),δ(ppm)28.3,50.5,61.0,61.6,63.6,66.0,67.0,69.9,70.5,70.6,72.2,72.6,98.7,119.0, 125.5,126.5,129.3；HRMS calcd for C₃₆H₄₃ClN₆O₅S[M+H]⁺:707.2777；found:707.2791.

如上所示的反应方程式为TC580的合成路线，反应条件具体如下：

(a)Pd(PPh₃)₄,Na₂CO₃,toluene/H₂O,90℃,3-5h；

(b)NaBH₄,MeOH,40℃,1h；

(c)HATU,DIPEA,DCM,40℃,12h；

(d)tetraethylene glycol,NaH(60％),THF,60℃,4-6h；

(e)DPPA,DBU,THF,r.t.to reflux,18-24h；

(f)2N HCl in dioxane,r.t.,2h.

在干燥的、充满氮气的反应瓶中加入2-氟-5-溴吡啶4.4g、3-甲酰基苯硼酸3.94g、四 (三苯基膦)钯577mg、碳酸氢钠6.1g、甲苯50mL和水100mL，加热至90℃搅拌5小时。反应结束后加入50mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝20:1)分离得到纯净的中间体54。

2.9g中间体54及3.0g 43溶于50mL甲醇中，加热至40℃搅拌0.5小时。向此溶液分批加入760mg硼氢化钠(NaBH₄)，继续在40℃搅拌0.5小时。反应结束后加入150mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝50:1)分离得到纯净的中间体55。¹H NMR(500MHz,CDCl₃),δ(ppm)1.26-1.48(m,4H,CH₂),1.46(s,9H,CH₃), 1.70-1.72(m,2H,CH₂),2.06-2.09(m,2H,CH₂),2.49-2.53(m,1H,CH),2.70(s,3H,CH₃),3.24 (br,1H,CH),3.89(s,2H,CH₂),6.99(dd,J＝8.5,3.0Hz,1H,CH),7.37-7.43(m,3H,CH),7.54(s, 1H,CH),7.99(td,J＝8.0,2.5Hz,1H,CH),8.40(d,J＝2.5Hz,1H,CH),8.24-8.26(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)28.1,28.2,31.7,50.5,53.2,79.0,109.1(d,J＝37.3Hz), 125.5,126.7,127.8,129.0,134.4(d,J＝4.5Hz),136.5,139.5(d,J＝8.0Hz),140.5,145.5(d,J＝14.6Hz),155.3,162.8(d,J＝237.8Hz)；HRMS calcd for C₂₄H₃₂FN₃O₂[M+H]⁺:414.2551；found: 414.2556.

2.55g 3-氯苯并[b]噻吩-2-甲酸及5.7g O-(7-氮苯并三氮唑)-N,N,N,N-四甲基脲六氟磷酸酯(HATU)溶于25mL DCM中，室温搅拌0.5小时。向此溶液加入4.13g中间体55和3.22g 二异丙基乙胺(DIPEA)的25mL DCM溶液，加热至40℃搅拌过夜。反应结束后加入50mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝3:1)分离得到纯净的中间体56。¹H NMR(500MHz,CDCl₃),δ(ppm)1.24-1.46(m,11H,CH₂and CH₃),1.64-1.74(m,5H,CH₂and CH),1.87-1.97(m,2H,CH₂),2.58 and 2.71(s,3H,CH₃),3.78-3.87(m,2H,CH₂),4.68and 4.84(s,2H,CH₂),7.01-7.02(m,1H,CH),7.22-7.62(m,6H,CH),7.72-8.01(m,3H,CH), 8.26-8.45(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)14.1,20.9,28.3,28.5,29.5,30.8, 45.1,52.5,53.4,58.6,60.3,79.4,109.2(d,J＝37.4Hz),118.9,122.6(d,J＝31.8Hz),125.4,125.6, 126.4,129.3,136.8,137.1,139.7,145.7(d,J＝14.4Hz),155.3,163.0(d,J＝237.9Hz),163.5, 171.0；HRMS calcd for C₃₃H₃₅ClFN₃O₃S[M+Na]⁺:630.1964；found:630.1958.

在582mg四乙二醇的10mL THF溶液中加入60mg NaH(60％)，室温搅拌15分钟，然后加入中间体56，加热至60℃反应6小时。反应结束后加入20mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝1:1)分离得到纯净的中间体57。HRMS calcd forC₄₁H₅₂ClN₃O₈S[M+H]⁺:782.3236；found:782.3226.

391mg中间体57、550mg叠氮磷酸二苯酯(DPPA)及228mg 1,8-二氮杂二环十一碳-7- 烯(DBU)溶于5mL THF中，室温搅拌6-12小时，通过TLC检测57完全消失后，继续加热回流12小时。反应结束后加入20mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝3:1)分离得到纯净的中间体58。¹H NMR(600MHz,CDCl₃),δ(ppm) 1.26-1.49(m,11H,CH₃and CH₂),1.59-1.96(m,6H,CH₂),2.56-2.70(s,4H,CH₃and CH), 3.50-3.68(m,2H,CH₂),3.64-3.74(m,12H,CH₂),3.88-3.89(m,2H,CH₂),4.54(br,2H,CH₂),4.66 and 4.84(s,2H,CH₂),6.84-6.88(m,1H,CH),7.10-7.52(m,8H,CH),7.61-7.89(m,4H,CH), 8.26-8.40(m,1H,CH)；¹³C NMR(150MHz,CDCl₃),δ(ppm)28.0,29.1,30.4,31.0,33.8,36.0, 40.0,42.4,44.8,50.2,58.4,64.9,69.2,69.6,70.21,70.24,70.9,79.0,110.8,118.6,119.8,120.0, 120.2,122.1,122.3,122.4,124.0,124.7,125.1,126.1,128.8,129.0,129.2,129.6,129.7,135.2, 136.8,137.0,137.1,137.7,144.4,155.0,162.1,162.6,163.1；HRMScalcd for C₄₁H₅₁ClN₆O₇S [M+H]⁺:807.3301；found:807.3313.

在150mg中间体58的2mL二氧六环溶液中加入2mL 4M HCl的二氧六环溶液，室温搅拌2小时。反应结束后浓缩、柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝30:1) 分离得到纯净的中间体TC580。¹H NMR(600MHz,CDCl₃),δ(ppm)1.13-1.45(m,2H,CH₂), 1.74-2.15(m,6H,CH₂),2.50and 2.60(s,3H,CH₃),2.81-2.94(m,1H,CH),3.27-3.31(m,3H,CH₂and CH),3.56-3.68(m,10H,CH₂),3.74-3.84(m,3H,CH₂and CH),4.43 and 4.64(s,2H,CH₂), 6.80-7.22(m,3H,CH),7.30-7.62(m,6H,CH),7.76-7.95(m,2H,CH),8.14-8.35(m,1H,CH)；¹³C NMR(150MHz,CDCl₃),δ(ppm)28.6,29.1,30.2,30.7,35.3,43.9,46.2,51.7,57.2,59.9,66.8, 70.6,71.0,71.46,71.58,71.64,112.1,120.0,121.0,121.3,121.6,123.3,124.2,124.6,126.0,126.4, 126.9,127.1,128.2,130.4,130.6,130.7,131.1,131.5,133.0,136.6,138.5,139.0,139.3,140.2, 145.5,162.4,162.6,164.5,165.5；HRMScalcd for C₃₆H₄₃ClN₆O₅S[M+H]⁺:707.2777；found: 707.2786.

如上所示的反应方程式为TC578的合成路线，反应条件具体如下：

(a)tetraethylene glycol,NaH(60％),DMF,120℃,12h；

(b)DPPA,DBU,THF,r.t.to reflux,18-24h.

在582mg四乙二醇的10mL DMF溶液中加入60mg NaH(60％)，室温搅拌15分钟，然后加入512mg LY2940680，加热至120℃反应12小时。反应结束后加入20mL饱和氯化铵溶液淬灭反应，用二氯甲烷萃取三次，有机相依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300 目硅胶，洗脱剂为二氯甲烷：甲醇＝20:1)分离得到纯净的中间体59。¹H NMR(500MHz, CDCl₃)majorrotamer,(ppm)1.76-2.26(m,4H,CH₂),2.77(s,3H,CH₃),3.37-3.39(m,2H, CH₂),3.61(d,J＝4.0Hz,2H,CH₂),3.67-3.75(m,12H,CH₂),3.88-3.91(m,2H,CH₂),4.06(s,3H, CH₃),4.20-4.24(m,4H,CH₂),4.90-4.94(m,1H,CH),6.60(d,J＝2.0Hz,1H,CH),7.17(d,J＝ 1.5Hz,1H,CH),7.25-7.39(m,2H,CH),7.66(d,J＝2.0Hz,1H,CH),7.82-7.91(m,2H,CH), 8.03-8.14(m,2H,CH)；minor rotamer,(ppm)1.76-2.24(m,4H,CH₂),3.10(s,3H,CH₃), 2.93-2.99(m,2H,CH₂),3.49-3.52(m,1H,CH),3.61(d,J＝4.0Hz,2H,CH₂),3.67-3.75(m,12H, CH₂),3.88-3.91(m,2H,CH₂),4.02(s,3H,CH₃),4.11-4.18(m,4H,CH₂),6.58(d,J＝2.0Hz,1H, CH),7.15(d,J＝1.5Hz,1H,CH),7.25-7.39(m,2H,CH),7.64(d,J＝2.0Hz,1H,CH),7.82-7.91 (m,2H,CH),8.03-8.14(m,2H,CH)；¹³C NMR(125MHz,CDCl₃)major rotamer,20.9,27.4, 27.9,28.5,31.6,38.0,50.7,51.0,56.7,61.4,67.7,69.2,70.0,70.3,70.4,70.6,72.4,108.9,112.8(q, J＝4.5Hz),118.0,121.2,123.6(q,J＝272.2Hz),124.5,124.8(q,J＝2.5Hz),126.0,127.4(q,J＝ 31.4Hz),127.7,128.4,131.4,131.9,136.6,138.0,147.2,158.7,159.4,168.8,171.0；minor rotamer, (ppm)14.0,29.0,29.5,38.0,49.9,50.2,50.9,53.3,60.2,67.7,69.2,70.0,70.3,70.4,70.6, 72.4,109.0,113.0(q,J＝4.5Hz),117.9,121.2,123.6(q,J＝272.2Hz),124.0(q,J＝2.1Hz), 124.3,126.0,127.4(q,J＝31.4Hz),127.4,128.0,131.5,132.0,136.5,138.0,147.5,158.8,159.3, 168.7,171.0；HRMS calcdfor C₃₄H₄₁F₃N₆O₆[M+H]⁺:687.3112；found:687.3135.

343mg中间体59、550mg叠氮磷酸二苯酯(DPPA)及228mg 1,8-二氮杂二环十一碳 -7-烯(DBU)溶于5mLTHF中，室温搅拌6-12小时，通过TLC检测中间体59完全消失后，继续加热回流12小时。反应结束后加入20mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝3:1)分离得到纯净的中间体TC578。¹H NMR(500MHz,CDCl₃) major rotamer,(ppm)1.63-1.66(m,2H,CH₂),1.74-2.27(m,4H,CH₂),2.75(s,3H,CH₃), 3.38-3.40(m,4H,CH₂),3.66-3.74(m,10H,CH₂),3.88-3.91(m,2H,CH₂),4.06(s,3H,CH₃), 4.19-4.22(m,4H,CH₂),4.90-4.96(m,1H,CH),6.59(d,J＝2.0Hz,1H,CH),7.14(d,J＝1.5Hz, 1H,CH),7.24-7.27(m,2H,CH),7.66(d,J＝2.0Hz,1H,CH),7.80-7.89(m,2H,CH),8.04-8.12 (m,2H,CH)；minor rotamer,(ppm)1.63-1.66(m,2H,CH₂),1.76-2.24(m,4H,CH₂),3.10(s,3H, CH₃),2.91-2.98(m,2H,CH₂),3.47-3.54(m,1H,CH),3.66-3.74(m,10H,CH₂),3.88-3.91(m,2H,CH₂),4.02(s,3H,CH₃),4.19-4.22(m,4H,CH₂),6.58(d,J＝2.0Hz,1H,CH),7.13(d,J＝1.5Hz, 1H,CH),7.24-7.27(m,2H,CH),7.65(d,J＝2.0Hz,1H,CH),7.88-7.89(m,2H,CH),8.04-8.12 (m,2H,CH)；¹³C NMR(125MHz,CDCl₃)major rotamer,27.2,27.7,28.4,28.8,29.2,31.4, 37.8,42.4,49.8,50.2,50.4,50.6,50.8,56.4,67.6,69.0,69.6,70.20,70.2170.24,70.4,70.9,108.7, 112.7(q,J＝4.5Hz),117.7,121.0,123.1(q,J＝272.2Hz),124.9,125.0(q,J＝2.5Hz),126.4, 127.4(q,J＝31.4Hz),127.7,128.3,129.4,131.2,131.7,136.4,137.7,147.0,158.5,159.2,168.6； minor rotamer,(ppm)14.0,29.0,29.5,37.8,49.9,50.2,50.9,53.3,60.2,67.7,69.2,70.0, 70.3,70.4,70.6,72.4,109.0,112.9(q,J＝4.5Hz),117.6,120.9,123.1(q,J＝272.2Hz),124.0(q,J ＝2.1Hz),124.3,126.3,127.4(q,J＝31.4Hz),127.4,128.3,129.2,131.3,131.8,136.3,137.7, 147.2,158.7,159.0,168.4；HRMS calcd for C₃₄H₄₀F₃N₉O₅[M+H]⁺:712.3177；found:712.3178.

如上所示的反应方程式为TC1306的合成路线，反应条件具体如下：

(a)SOCl₂,70℃,6h；

(b)i.thiourea,H₂O,reflux,18h；ii.3M NaOH,reflux,3h；iii.HCl；

(c)K₂CO₃,DMF,r.t.,12h；

(d)DPPA,DBU,THF,r.t.to reflux,18-24h；

(e)aq.NaOH,reflux,12h；

(f)mCPBA,DCM,r.t.,1h；

(g)SnCl₂,EtOH,pH<3,reflux,6h；

(h)HATU,DIPEA,DCM,r.t.,12h.

合成TC1306及同系物的通用步骤：

3.0g三乙二醇溶于20mL氯化亚砜(SOCl₂)中，加热至70℃搅拌6小时。反应结束后浓缩得到粗产物，用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝1:1)分离得到纯净的中间体60。

1.68g中间体60及1.06g硫脲溶于30mL水中，加热至回流搅拌18小时，然后加入3MNaOH溶液10mL，继续回流搅拌3小时。反应结束后加入盐酸中和反应，浓缩除去水得到粗产物，柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝20:1)分离纯化得到无色液体中间体64。

830mg中间体64、775mg 2-氯-4-氟苯腈及1.06g碳酸钾溶于30mL DMF中，室温搅拌12小时。反应结束后加入100mL饱和氯化铵溶液淬灭反应，用二氯甲烷萃取三次，有机相依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝1:1)分离得到纯净的中间体68。¹H NMR(500MHz,CDCl₃),δ(ppm)3.22(t,J＝5.0Hz,2H,CH₂), 3.59-3.61(m,2H,CH₂),3.66(s,4H,CH₂),3.73-3.77(m,4H,CH₂),7.23-7.25(m,1H,CH),7.39(s, 1H,CH),7.52(d,J＝5.0Hz,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)31.7,61.6,69.2,70.2,70.6,72.5,109.0,116.1,124.9,127.0,133.6,137.0,146.3；HRMS calcd for C₁₃H₁₆ClNO₃S[M+H]⁺: 302.0612.

中间体69。¹H NMR(500MHz,CDCl₃),δ(ppm)3.22(t,J＝6.5Hz,2H,CH₂),3.61(t,J＝5.0Hz,2H,CH₂),3.64-3.69(m,8H,CH₂),3.72-3.76(m,4H,CH₂),7.25(dd,J＝8.5,2.0Hz,1H,CH),7.41(d,J＝2.0Hz,1H,CH),7.52(d,J＝8.0Hz,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)31.5,61.5,69.2,70.1,70.3,70.46,70.52,72.5,108.9,116.0,124.8,126.9,133.5,136.9, 146.3；HRMS calcd for C₁₅H₂₀ClNO₄S[M+Na]⁺:368.0694；found:368.0696.

中间体70。¹H NMR(500MHz,CDCl₃),δ(ppm)3.22(t,J＝5.0Hz,2H,CH₂),3.60-3.75(m, 18H,CH₂),7.26(dd,J＝5.0,10.0Hz,1H,CH),7.41(d,J＝5.0Hz,1H,CH),7.53(d,J＝5.0Hz, 1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)31.5,61.6,69.4,70.1,70.4,70.52,70.58,70.6, 72.7,109.0,116.1,124.9,127.1,133.6,137.0,146.4；HRMS calcd forC₁₇H₂₄ClNO₅S[M+Na]⁺: 412.0956；found:412.0951.

中间体71。¹H NMR(500MHz,CDCl₃),δ(ppm)3.20-3.24(m,2H,CH₂),3.59-3.77(m,22H, CH₂),7.26(d,J＝5.0Hz,1H,CH),7.41(s,1H,CH),7.53(dd,J＝10.0,5.0Hz,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)31.6,61.5,69.2,70.1,70.35,70.41,70.44,72.5,108.8,116.0, 124.8,126.8,133.4,136.8,146.4；HRMS calcd for C₁₉H₂₈ClNO₆S[M+Na]⁺:456.1218；found: 456.1267.

150mg中间体68、550mg叠氮磷酸二苯酯(DPPA)及228mg 1,8-二氮杂二环十一碳 -7-烯(DBU)溶于5mL THF中，室温搅拌6-12小时，通过TLC检测中间体68完全消失后，继续加热回流12小时。反应结束后加入20mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝1:1)分离得到纯净的中间体72。¹H NMR(500MHz,CDCl₃),δ(ppm) 3.20(t,J＝5.0Hz,2H,CH₂),3.38(t,J＝5.0Hz,2H,CH₂),3.64-3.68(m,4H,CH₂),3.76(t,J＝5.0 Hz,2H,CH₂),5.18(d,J＝10.0Hz,2H,CH₂),7.20-7.25(m,1H,CH),7.33-7.40(m,1H,CH),7.40 (d,J＝5.0Hz,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)115.3,120.09,120.13,120.6,125.6,129.6,129.9,150.36,150.42；HRMS calcd for C₁₃H₁₅ClN₄O₂S[M+Na]⁺:349.0496；found:349.0472.

中间体73。¹H NMR(500MHz,CDCl₃),δ(ppm)3.17-3.22(m,2H,CH₂),3.34-3.40(m,2H, CH₂),3.62-3.68(m,10H,CH₂),3.71-3.77(m,2H,CH₂),7.20-7.25(m,1H,CH),7.36-7.40(m,1H, CH),7.48-7.53(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)31.6,50.5,69.2,69.8,70.4, 108.8,115.9,124.8,126.8,133.4,136.8,146.4；HRMS calcd for C₁₅H₁₉ClN₄O₃S[M+Na]⁺: 393.0759；found:393.0756.

中间体74。¹H NMR(500MHz,CDCl₃),δ(ppm)2.91-2.98(m,4H,CH₂),3.14-3.19(m,2H, CH₂),3.32-3.36(m,2H,CH₂),3.60-3.63(m,10H,CH₂),3.68-3.73(m,2H,CH₂),7.19-7.29(m,1H, CH),7.35-7.37(m,1H,CH),7.46-7.49(m,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)31.3, 33.7,39.9,50.2,68.8,69.5,70.1,108.3,115.6,120.1,124.0,124.5,126.4,129.0,133.1,136.3, 146.3；HRMS calcd for C₁₇H₂₃ClN₄O₄S[M+Na]⁺:437.1021；found:437.1008.

中间体75。¹H NMR(500MHz,CDCl₃),δ(ppm)3.16-3.23(m,2H,CH₂),3.34-3.38(m,2H, CH₂),3.61-3.76(m,20H,CH₂),7.08-7.12(m,1H,CH),7.37-7.38(m,1H,CH),7.48-7.52(m,1H, CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)31.3,33.8,40.0,50.2,68.9,69.5,70.05,70.13,108.3, 115.6,120.1,123.9,124.5,126.4,129.0,133.1,136.4,146.3,151.0；HRMScalcd for C₁₉H₂₇ClN₄O₅S[M+Na]⁺:481.1283；found:481.1247.

652mg中间体72溶于10mL 1M NaOH水溶液中，加热回流反应12小时。反应结束后加入盐酸中和反应，浓缩除去水得到粗产物，柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝20:1)分离得到纯净的中间体76。¹H NMR(500MHz,CDCl₃),δ(ppm)3.21(t,J＝5.0Hz,2H,CH₂),3.39(t,J＝5.0Hz,2H,CH₂),3.67-3.69(m,6H,CH₂),3.76(t,J＝5.0Hz,2H,CH₂),7.22 (dd,J＝5.0,10.0Hz,1H,CH),7.37(d,J＝5.0Hz,1H,CH),7.91(d,J＝5.0Hz,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)31.7,50.6,69.5,70.1,70.6,124.7,124.9,128.7,132.6,135.3, 145.0,169.7；HRMS calcd for C₁₃H₁₆ClN₃O₄S[M+Na]⁺:368.0442；found:368.0432.

中间体77。¹H NMR(500MHz,CDCl₃),δ(ppm)3.17(t,J＝6.5Hz,2H,CH₂),3.38(t,J＝5.0Hz,2H,CH₂),3.66-3.67(m,10H,CH₂),3.72(t,J＝6.5Hz,2H,CH₂),7.14(d,J＝8.0Hz,1H,CH),7.29-7.30(m,1H,CH),7.79(d,J＝8.5Hz,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)31.6,50.4,69.3,69.8,70.3,70.38,70.41,70.42,124.6,126.6,128.5,132.1,134.4,143.5,169.5； HRMS calcd for C₁₅H₂₀ClN₃O₅S[M+Na]⁺:412.0704；found:412.0712.

中间体78。¹H _NMR(500MHz,CDCl₃),δ(ppm)3.15-3.23(m,2H,CH₂),3.37-3.41(m,2H,CH₂),3.68-3.76(m,16H,CH₂),7.19-7.20(m,1H,CH),7.34-7.36(m,1H,CH),7.87-7.90(m,1H, CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)31.6,50.6,69.4,69.9,70.4,70.5,113.1,124.6,125.1, 128.6,132.5,135.0,144.6,151.4,168.3；HRMS calcd for C₁₇H₂₄ClN₃O₆S[M+Na]⁺:456.0967； found:456.0925.

中间体79。¹H NMR(500MHz,CDCl₃),δ(ppm)3.23-3.72(m,24H,CH₂),7.34-7.90(m,3H, CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)31.7,50.7,54.0,70.5,124.7,128.7,137.4；HRMScalcd for C₁₉H₂₈ClN₃O₇S[M+Na]⁺:500.1229；found:500.1207.

518mg中间体76溶于10mL DCM中，加入776mg间氯过氧苯甲酸(mCPBA)，室温搅拌1小时。反应结束后浓缩除去溶剂得到粗产物，柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝10:1)分离得到纯净的中间体80。¹H NMR(500MHz,CDCl₃),δ(ppm)3.35(t,J ＝5.0Hz,2H,CH₂),3.48-3.59(m,8H,CH₂),3.91(t,J＝10.0Hz,2H,CH₂),7.92(dd,J＝10.0,5.0 Hz,1H,CH),8.06(d,J＝2.0Hz,1H,CH),8.11(d,J＝10.0Hz,1H,CH)；¹³C NMR(125MHz, CDCl₃),δ(ppm)50.5,56.4,64.5,70.0,70.2,70.5,126.2,130.9,132.5,133.5,135.0,144.5,168.3；HRMS calcd for C₁₃H₁₆ClN₃O₆S[M+Na]⁺:400.0341；found:400.0318.

中间体81。¹H NMR(500MHz,CDCl₃),δ(ppm)3.58-3.42(m,4H,CH₂),3.50(br,4H,CH₂), 3.57-3.62(m,6H,CH₂),3.82-3.84(m,2H,CH₂),7.64-7.65(m,1H,CH),7.79(br,2H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)50.5,55.9,64.1,69.7,70.0,70.2,70.3,126.0,129.5,130.6, 131.6,132.5,141.3；HRMS calcd for C₁₅H₂₀ClN₃O₇S[M+Na]⁺:444.0603；found:444.0598.

中间体82。¹H NMR(500MHz,CDCl₃),δ(ppm)3.41(t,J＝5.0Hz,2H,CH₂),3.46-3.69(m, 16H,CH₂),3.89(t,J＝5.0Hz,2H,CH₂),7.90(dd,J＝10.0,10.0Hz,1H,CH),8.04-8.07(m,2H, CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm)50.6,56.4,64.5,69.9,70.1,70.4,70.5,70.6,126.3, 130.8,132.3,134.7,144.2,167.2；HRMS calcd for C₁₇H₂₄ClN₃O₈S[M+Na]⁺:488.0865；found: 488.0855.

中间体83。¹H NMR(500MHz,CDCl₃),δ(ppm)3.40-3.69(m,22H,CH₂),3.88(t,J＝5.0Hz,2H,CH₂),7.88(d,J＝10.0Hz,1H,CH),8.02-8.05(m,2H,CH)；¹³C NMR(125MHz,CDCl₃), δ(ppm)50.5,56.3,64.4,69.8,70.0,70.2,70.28,70.32,70.36,70.4,126.2,130.5,132.0,134.3, 134.7,143.7,166.5；HRMS calcd for C₁₉H₂₈ClN₃O₉S[M+Na]⁺:532.1327.

在351mg 2-(2-氯-5-硝基苯基)吡啶的10mL乙醇溶液中加入盐酸调节pH<3，加入1.35g 二水合氯化亚锡，加热至回流搅拌6小时。反应结束后降至0℃，加入NaOH溶液调节pH>12，用硅藻土过滤除去沉淀，浓缩除去乙醇，用乙酸乙酯萃取3次，有机相依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝1:1)分离得到纯净的中间体84。

83mg 80及117mg HATU溶于5mLDCM中，室温搅拌0.5小时。向此溶液加入40mg 84和71mg DIPEA的3mL DCM溶液，室温搅拌12小时。反应结束后加入20mL饱和氯化铵溶液淬灭反应，分出有机相，水相用二氯甲烷萃取三次，并入有机相，依次用饱和氯化铵水溶液和饱和食盐水洗涤，然后用无水硫酸钠干燥，过滤除去硫酸钠，浓缩得到粗产品。粗产品用柱层析(200-300目硅胶，洗脱剂为正己烷：乙酸乙酯＝1:1)分离得到纯净的中间体 TC1306。¹HNMR(500MHz,CDCl₃),δ(ppm)3.16-3.21(m,4H,CH₂),3.33(t,J＝5.0Hz,2H, CH₂),3.42(t,J＝5.0Hz,2H,CH₂),3.47-3.52(m,4H,CH₂),3.57(t,J＝5.0Hz,2H,CH₂), 3.68-3.74(m,4H,CH₂),3.86(t,J＝5.0Hz,2H,CH₂),7.31-7.34(m,1H,CH),7.48(d,J＝10.0Hz, 1H,CH),7.70-7.95(m,6H,CH),7.59(d,J＝10.0Hz,1H,CH),8.95(s,1H,CH)；¹³C NMR(125 MHz,CDCl₃),δ(ppm).50.6,55.8,56.5,64.6,70.1,70.3,70.5,123.1,130.0；HRMS calcd forC2₅H₂₃Cl₂N₅O₅S[M+Na]⁺:586.0689；found:586.0722.

TC1156。¹H NMR(500MHz,CDCl₃),δ(ppm)3.46(t,J＝5.0Hz,2H,CH₂),3.52(t,J＝5.5 Hz,2H,CH₂),3.58(br,4H,CH₂),3.62-3.66(m,6H,CH₂),3.92(t,J＝5.5Hz,2H,CH₂),7.37-7.41 (m,1H,CH),7.50(d,J＝9.0Hz,1H,CH),7.70-7.71(m,1H,CH),7.75-7.76(m,2H,CH), 7.85-7.88(m,2H,CH),7.92-7.94(m,1H,CH),8.03(d,J＝1.5Hz,1H,CH),8.62-8.63(m,1H, CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm).50.3,55.8,63.9,69.3,69.6,69.82,69.84,70.0,121.6, 122.6,122.8,125.1,126.4,127.4,129.3,129.4,130.3,131.9,136.6,138.6,140.8,141.6,148.5, 155.8,164.2；HRMS calcd for C₂₆H₂₇Cl₂N₅O₆S[M+H]⁺:608.1132；found:608.1126.

TC1307。¹H NMR(500MHz,CDCl₃),δ(ppm)3.34-3.66(m,18H,CH₂),3.82-3.85(m,2H,CH₂),7.24-7.34(m,2H,CH),7.48(dd,J＝10.0,10.0Hz,1H,CH),7.61-7.97(m,6H,CH),8.54(d, J＝5.0Hz,1H,CH)；¹³C NMR(125MHz,CDCl₃),δ(ppm).50.6,56.3,56.4,64.3,64.4,69.8,69.9, 70.0,70.1,70.4,70.5,121.6,122.8,122.9,125.1,129.7,130.7,136.3；HRMScalcd for C2₂₉H₃₁Cl₂N₅O₇S[M+Na]⁺:674.1213；found:674.1216.

TC820。¹H NMR(600MHz,CDCl₃),δ(ppm)2.80(br,2H,CH₂),3.35(t,J＝5.4Hz,2H,CH₂),3.40(t,J＝5.4Hz,2H,CH₂),3.44-3.49(m,4H,CH₂),3.53-3.54(m,2H,CH₂),3.58-3.62(m, 10H,CH₂),3.82(t,J＝6.0Hz,2H,CH₂),7.30-7.32(m,1H,CH),7.46(d,J＝9.0Hz,1H,CH), 7.66-7.71(m,2H,CH),7.77-7.84(m,3H,CH),7.90-7.91(m,2H,CH),8.52(br,1H,CH),9.71(s, 1H,CH)；¹³C NMR(150MHz,CDCl₃),δ(ppm).50.5,56.4,64.4,69.8,70.0,70.29,70.34,70.36, 70.43,70.47,121.8,122.9,123.0,125.4,126.7,127.3,129.6,129.7,130.7,131.8,136.96,136.99, 138.2,140.3,142.3,148.5,155.5,163.9；HRMS calcd forC₃₀H₃₅Cl₂N₅O₈S[M+Na]⁺:718.1481； found:718.1423.

实施例2

平滑受体的双头配体通用合成方法描述如下：

将两个中间体分子各0.1mmol溶于3甲醇中，加入1M硫酸铜水溶液0.4mL及1M抗坏血酸钠水溶液0.4mL，室温搅拌1小时。反应结束后浓缩、柱层析(200-300目硅胶，洗脱剂为二氯甲烷：甲醇＝10:1至5:1不等)分离纯化得到相应的双头分子。

合成每一个双头配体对应的两个片段分子编号及反应产率如表1所示：

表1

将各配体进行结构表征，具体如下：

TC802:¹H NMR(800MHz,CDCl₃),(ppm)0.83(s,3H,CH₃),0.87-0.97(m,1H,CH₂),0.98(s, 3H,CH₃),1.02-1.22(m,4H,CH₂and CH),1.23(s,3H,CH₃),1.25-1.74(m,20H,CH₂and CH), 1.80-1.82(m,2H,CH₂),1.93-1.96(m,1H,CH₂),2.04-2.06(m,1H,CH₂),2.20-2.28(m,2H,CH₂), 3.42-3.50(m,3H,CH₂and CH),4.46-4.48(m,4H,CH₂),4.55(s,2H,CH₂),4.77-4.78(m,1H,CH), 5.31-5.32(m,1H,CH),6.99-7.00(m,1H,CH),7.21-7.27(m,1H,CH),7.40-7.51(m,6H,CH), 7.75-7.87(m,2H,CH)；¹³C NMR(200MHz,CDCl₃),13.5,19.3,20.8,22.3,23.7,24.0,25.9, 26.2,29.4,29.9,31.2,31.5,31.7,36.4,37.2,40.0,42.1,42.5,43.8,47.9,49.9,56.8,57.5,63.9,70.6, 71.5,75.0,119.2,121.3,122.7,123.8,125.6,126.7,128.2,129.3,137.0,140.8,145.5,163.7； HRMS calcd for C₆₀H₇₉ClN₆O₅S[M+H]⁺:1031.5594；found:1031.5553.

TC803:¹H NMR(800MHz,CDCl₃),(ppm)0.84(s,3H,CH₃),0.87-0.98(m,1H,CH₂),0.99(s, 3H,CH₃),1.02-1.19(m,4H,CH₂and CH),1.24(s,3H,CH₃),1.25-1.75(m,20H,CH₂and CH), 1.80-1.85(m,2H,CH₂),1.94-1.97(m,1H,CH₂),2.04-2.07(m,1H,CH₂),2.20-2.28(m,2H,CH₂), 3.46-3.50(m,3H,CH₂and CH),3.83-3.84(m,2H,CH₂),3.93-3.94(m,2H,CH₂),4.50-4.51(m, 2H,CH₂),4.55-4.57(s,4H,CH₂),4.82(br,1H,CH),5.32-5.33(m,1H,CH),6.96-7.24(m,3H, CH),7.45-7.52(m,5H,CH),7.68-7.86(m,3H,CH),8.17(br,1H,CH)；¹³C NMR(200MHz, CDCl₃),13.5,19.3,20.8,22.3,23.7,24.1,26.0,26.3,29.5,30.0,31.2,31.5,31.7,36.4,37.2, 40.0,42.2,42.5,43.8,50.0,50.2,56.8,57.5,64.1,64.9,69.5,69.8,70.6,71.5,75.0,108.6,119.0, 121.4,122.7,123.6,125.4,126.5,127.4,129.2,137.1,140.8,145.2,147.1,164.1；HRMS calcd for C₆₂H₈₃ClN₆O₆S[M+2H]²⁺:538.2964；found:538.2955.

TC804:¹H NMR(800MHz,CDCl₃),(ppm)0.85(s,3H,CH₃),0.87-0.98(m,1H,CH₂),1.00(s, 3H,CH₃),1.04-1.21(m,4H,CH₂and CH),1.25(s,3H,CH₃),1.26-1.75(m,20H,CH₂and CH), 1.82-1.84(m,2H,CH₂),1.95-2.02(m,1H,CH₂),2.07-2.08(m,1H,CH₂),2.20-2.29(m,2H,CH₂), 3.42-3.52(m,3H,CH₂and CH),3.56-3.70(m,8H,CH₂),3.85-3.87(m,4H,CH₂),4.51-4.53(m, 4H,CH₂),4.66(s,2H,CH₂),4.82(br,1H,CH),5.33-5.34(m,1H,CH),7.01-7.26(m,2H,CH), 7.45-7.50(m,4H,CH),7.70-7.87(m,3H,CH),8.17(br,1H,CH)；¹³CNMR(200MHz,CDCl₃), 13.6,19.4,20.9,22.3,23.7,24.2,26.0,26.4,29.6,30.0,31.3,31.6,31.7,36.4,37.2,40.1,42.2, 42.6,43.9,50.0,50.2,56.8,57.6,64.5,65.1,69.5,69.6,69.8,70.0,70.46,70.51,70.57,70.58,71.4, 71.6,75.1,108.7,119.0,121.5,122.7,123.8,125.4,126.5,127.4,129.2,137.2,140.8,144.9,147.1, 164.1；HRMS calcdfor C₆₄H₈₇ClN₆O₇S[M+2H]²⁺:560.3096；found:560.3088.

TC593:¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H,CH₃),0.88-0.99(m,1H,CH₂),1.01(s, 3H,CH₃),1.02-1.25(m,4H,CH₂and CH),1.26(s,3H,CH₃),1.27-1.78(m,20H,CH₂and CH), 1.82-1.86(m,2H,CH₂),1.95-2.00(m,1H,CH₂),2.07-2.11(m,1H,CH₂),2.20-2.32(m,2H,CH₂), 3.46-3.51(m,3H,CH₂and CH),3.59-3.74(m,8H,CH₂),3.84-3.90(m,4H,CH₂),4.50-4.54(m, 4H,CH₂),4.60(s,2H,CH₂),4.82(br,1H,CH),5.31-5.32(m,1H,CH),6.84-7.15(m,3H,CH), 7.45-7.50(m,5H,CH),7.67-7.88(m,3H,CH),8.17(br,1H,CH)；¹³CNMR(125MHz,CDCl₃), 13.4,19.2,20.7,22.2,23.6,24.0,26.0,26.2,29.4,30.0,31.1,31.4,31.6,36.3,37.1,39.9,42.2, 42.4,43.8,44.9,49.8,50.0,53.4,56.7,57.4,59.1,64.0,65.1,69.3,69.6,70.33,70.35,70.45,70.50, 70.56,70.59,71.3,74.9,108.6,115.4,118.9,121.3,122.4,122.6,123.5,125.2,125.4,126.4,127.3, 129.1,137.0,140.8,145.0,147.0,163.6,164.1；HRMS calcd for C₆₆H₉₁ClN₆O₈S[M+2H]²⁺: 582.3227；found:582.3217.

TC594:¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H,CH₃),0.88-0.99(m,1H,CH₂),1.01(s, 3H,CH₃),1.02-1.25(m,4H,CH₂and CH),1.26(s,3H,CH₃),1.27-1.78(m,20H,CH₂and CH), 1.82-1.86(m,2H,CH₂),1.95-2.00(m,1H,CH₂),2.07-2.11(m,1H,CH₂),2.20-2.32(m,2H,CH₂), 3.46-3.51(m,3H,CH₂and CH),3.59-3.74(m,8H,CH₂),3.84-3.90(m,4H,CH₂),4.50-4.54(m, 4H,CH₂),4.60(s,2H,CH₂),4.82(br,1H,CH),5.31-5.32(m,1H,CH),6.84-7.15(m,3H,CH), 7.45-7.50(m,5H,CH),7.67-7.88(m,3H,CH),8.17(br,1H,CH)；¹³CNMR(125MHz,CDCl₃), 13.4,19.2,20.7,22.2,23.6,24.0,26.0,26.2,29.4,30.0,31.1,31.4,31.6,36.3,37.1,39.9,42.2, 42.4,43.8,44.9,49.8,50.0,53.4,56.7,57.4,59.1,64.0,65.1,69.3,69.6,70.33,70.35,70.45,70.50, 70.56,70.59,71.3,74.9,108.6,115.4,118.9,121.3,122.4,122.6,123.5,125.2,125.4,126.4,127.3, 129.1,137.0,140.8,145.0,147.0,163.6,164.1；HRMS calcd for C₆₈H₉₅ClN₆O₉S[M+2H]²⁺: 604.3358；found:604.3347.

TC595:¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H,CH₃),0.88-0.99(m,1H,CH₂),1.01(s, 3H,CH₃),1.02-1.25(m,4H,CH₂and CH),1.26(s,3H,CH₃),1.27-1.78(m,20H,CH₂and CH), 1.82-1.86(m,2H,CH₂),1.95-2.00(m,1H,CH₂),2.07-2.11(m,1H,CH₂),2.20-2.32(m,2H,CH₂), 3.46-3.51(m,3H,CH₂and CH),3.59-3.74(m,8H,CH₂),3.84-3.90(m,4H,CH₂),4.50-4.54(m, 4H,CH₂),4.60(s,2H,CH₂),4.82(br,1H,CH),5.31-5.32(m,1H,CH),6.84-7.15(m,3H,CH), 7.45-7.50(m,5H,CH),7.67-7.88(m,3H,CH),8.17(br,1H,CH)；¹³CNMR(125MHz,CDCl₃), 13.4,19.2,20.7,22.2,23.6,24.0,26.0,26.2,29.4,30.0,31.1,31.4,31.6,36.3,37.1,39.9,42.2, 42.4,43.8,44.9,49.8,50.0,53.4,56.7,57.4,59.1,64.0,65.1,69.3,69.6,70.33,70.35,70.45,70.50, 70.56,70.59,71.3,74.9,108.6,115.4,118.9,121.3,122.4,122.6,123.5,125.2,125.4,126.4,127.3, 129.1,137.0,140.8,145.0,147.0,163.6,164.1；HRMS calcd for C₇₀H₉₉ClN₆O₁₀S[M+2H]²⁺: 626.3489；found:626.3477.

TC596:¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H,CH₃),0.88-0.99(m,1H,CH₂),1.01(s, 3H,CH₃),1.02-1.25(m,4H,CH₂and CH),1.26(s,3H,CH₃),1.27-1.78(m,20H,CH₂and CH), 1.82-1.86(m,2H,CH₂),1.95-2.00(m,1H,CH₂),2.07-2.11(m,1H,CH₂),2.20-2.32(m,2H,CH₂), 3.46-3.51(m,3H,CH₂and CH),3.59-3.74(m,8H,CH₂),3.84-3.90(m,4H,CH₂),4.50-4.54(m, 4H,CH₂),4.60(s,2H,CH₂),4.82(br,1H,CH),5.31-5.32(m,1H,CH),6.84-7.15(m,3H,CH), 7.45-7.50(m,5H,CH),7.67-7.88(m,3H,CH),8.17(br,1H,CH)；¹³CNMR(125MHz,CDCl₃), 13.4,19.2,20.7,22.2,23.6,24.0,26.0,26.2,29.4,30.0,31.1,31.4,31.6,36.3,37.1,39.9,42.2, 42.4,43.8,44.9,49.8,50.0,53.4,56.7,57.4,59.1,64.0,65.1,69.3,69.6,70.33,70.35,70.45,70.50, 70.56,70.59,71.3,74.9,108.6,115.4,118.9,121.3,122.4,122.6,123.5,125.2,125.4,126.4,127.3, 129.1,137.0,140.8,145.0,147.0,163.6,164.1；HRMS calcd for C₇₂H₁₀₃ClN₆O₁₁S[M+2H]²⁺: 648.3620；found:648.3609.

TC589:¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H,CH₃),0.88-0.99(m,1H,CH₂),1.01(s, 3H,CH₃),1.02-1.25(m,4H,CH₂and CH),1.26(s,3H,CH₃),1.27-1.78(m,20H,CH₂and CH), 1.82-1.86(m,2H,CH₂),1.95-2.00(m,1H,CH₂),2.07-2.11(m,1H,CH₂),2.20-2.32(m,2H,CH₂), 3.46-3.51(m,3H,CH₂and CH),3.59-3.74(m,8H,CH₂),3.84-3.90(m,4H,CH₂),4.50-4.54(m, 4H,CH₂),4.60(s,2H,CH₂),4.82(br,1H,CH),5.31-5.32(m,1H,CH),6.84-7.15(m,3H,CH), 7.45-7.50(m,5H,CH),7.67-7.88(m,3H,CH),8.17(br,1H,CH)；¹³CNMR(125MHz,CDCl₃), 13.4,19.2,20.7,22.2,23.6,24.0,26.0,26.2,29.4,30.0,31.1,31.4,31.6,36.3,37.1,39.9,42.2, 42.4,43.8,44.9,49.8,50.0,53.4,56.7,57.4,59.1,64.0,65.1,69.3,69.6,70.33,70.35,70.45,70.50, 70.56,70.59,71.3,74.9,108.6,115.4,118.9,121.3,122.4,122.6,123.5,125.2,125.4,126.4,127.3, 129.1,137.0,140.8,145.0,147.0,163.6,164.1；HRMS calcd for C₇₄H₁₀₇ClN₆O₁₂S[M+2H]²⁺: 670.3751；found:670.3737.

TC597:¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H,CH₃),0.88-0.99(m,1H,CH₂),1.01(s, 3H,CH₃),1.02-1.25(m,4H,CH₂and CH),1.26(s,3H,CH₃),1.27-1.78(m,20H,CH₂and CH), 1.82-1.86(m,2H,CH₂),1.95-2.00(m,1H,CH₂),2.07-2.11(m,1H,CH₂),2.20-2.32(m,2H,CH₂), 3.46-3.51(m,3H,CH₂and CH),3.59-3.74(m,8H,CH₂),3.84-3.90(m,4H,CH₂),4.50-4.54(m, 4H,CH₂),4.60(s,2H,CH₂),4.82(br,1H,CH),5.31-5.32(m,1H,CH),6.84-7.15(m,3H,CH), 7.45-7.50(m,5H,CH),7.67-7.88(m,3H,CH),8.17(br,1H,CH)；¹³CNMR(125MHz,CDCl₃), 13.4,19.2,20.7,22.2,23.6,24.0,26.0,26.2,29.4,30.0,31.1,31.4,31.6,36.3,37.1,39.9,42.2, 42.4,43.8,44.9,49.8,50.0,53.4,56.7,57.4,59.1,64.0,65.1,69.3,69.6,70.33,70.35,70.45,70.50, 70.56,70.59,71.3,74.9,108.6,115.4,118.9,121.3,122.4,122.6,123.5,125.2,125.4,126.4,127.3, 129.1,137.0,140.8,145.0,147.0,163.6,164.1；HRMS calcd for C₇₆H₁₁₁ClN₆O₁₃S[M+2H]²⁺: 692.3882；found:692.3866.

TC598:¹H NMR(500MHz,CDCl₃),(ppm)0.86(s,3H,CH₃),0.88-0.99(m,1H,CH₂),1.01(s, 3H,CH₃),1.02-1.25(m,4H,CH₂and CH),1.26(s,3H,CH₃),1.27-1.78(m,20H,CH₂and CH), 1.82-1.86(m,2H,CH₂),1.95-2.00(m,1H,CH₂),2.07-2.11(m,1H,CH₂),2.20-2.32(m,2H,CH₂), 3.46-3.51(m,3H,CH₂and CH),3.59-3.74(m,8H,CH₂),3.84-3.90(m,4H,CH₂),4.50-4.54(m, 4H,CH₂),4.60(s,2H,CH₂),4.82(br,1H,CH),5.31-5.32(m,1H,CH),6.84-7.15(m,3H,CH), 7.45-7.50(m,5H,CH),7.67-7.88(m,3H,CH),8.17(br,1H,CH)；¹³CNMR(125MHz,CDCl₃), 13.4,19.2,20.7,22.2,23.6,24.0,26.0,26.2,29.4,30.0,31.1,31.4,31.6,36.3,37.1,39.9,42.2, 42.4,43.8,44.9,49.8,50.0,53.4,56.7,57.4,59.1,64.0,65.1,69.3,69.6,70.33,70.35,70.45,70.50, 70.56,70.59,71.3,74.9,108.6,115.4,118.9,121.3,122.4,122.6,123.5,125.2,125.4,126.4,127.3, 129.1,137.0,140.8,145.0,147.0,163.6,164.1；HRMS calcd for C₇₈H₁₁₅ClN₆O₁₄S[M+2H]²⁺: 714.4013；found:714.4000.

TC805:¹H NMR(800MHz,CDCl₃),(ppm)0.85(s,3H,CH₃),0.87-0.98(m,1H,CH₂),1.00(s, 3H,CH₃),1.04-1.21(m,4H,CH₂and CH),1.25(s,3H,CH₃),1.26-1.75(m,20H,CH₂and CH), 1.82-1.84(m,2H,CH₂),1.95-2.02(m,1H,CH₂),2.07-2.08(m,1H,CH₂),2.20-2.29(m,2H,CH₂), 3.42-3.52(m,3H,CH₂and CH),3.56-3.70(m,8H,CH₂),3.85-3.87(m,4H,CH₂),4.51-4.53(m, 4H,CH₂),4.66(s,2H,CH₂),4.82(br,1H,CH),5.33-5.34(m,1H,CH),7.01-7.26(m,3H,CH), 7.45-7.50(m,5H,CH),7.70-7.87(m,3H,CH),8.17(br,1H,CH)；¹³CNMR(200MHz,CDCl₃), 13.6,19.4,20.9,22.3,23.7,24.2,26.0,26.4,29.6,30.0,31.3,31.6,31.7,36.4,37.2,40.1,42.2, 42.6,43.9,50.0,50.2,56.8,57.6,64.5,65.1,69.5,69.6,69.8,70.0,70.46,70.51,70.57,70.58,71.4, 71.6,75.1,108.7,119.0,121.5,122.7,123.8,125.4,126.5,127.4,129.2,137.2,140.8,144.9,147.1, 164.1；HRMS calcdfor C₆₈H₉₅ClN₆O₉S[M+H]⁺:1207.6643；found:1207.6626.

TC806:¹H NMR(800MHz,CDCl₃),(ppm)0.85(s,3H,CH₃),0.87-0.98(m,1H,CH₂),1.00(s, 3H,CH₃),1.04-1.21(m,4H,CH₂and CH),1.25(s,3H,CH₃),1.26-1.75(m,20H,CH₂and CH), 1.82-1.84(m,2H,CH₂),1.95-2.02(m,1H,CH₂),2.07-2.08(m,1H,CH₂),2.20-2.29(m,2H,CH₂), 3.42-3.52(m,3H,CH₂and CH),3.56-3.70(m,8H,CH₂),3.85-3.87(m,4H,CH₂),4.51-4.53(m, 4H,CH₂),4.66(s,2H,CH₂),4.82(br,1H,CH),5.33-5.34(m,1H,CH),7.01-7.26(m,2H,CH), 7.45-7.50(m,4H,CH),7.70-7.87(m,3H,CH),8.17(br,1H,CH)；¹³CNMR(200MHz,CDCl₃), 13.6,19.4,20.9,22.3,23.7,24.2,26.0,26.4,29.6,30.0,31.3,31.6,31.7,36.4,37.2,40.1,42.2, 42.6,43.9,50.0,50.2,56.8,57.6,64.5,65.1,69.5,69.6,69.8,70.0,70.46,70.51,70.57,70.58,71.4, 71.6,75.1,108.7,119.0,121.5,122.7,123.8,125.4,126.5,127.4,129.2,137.2,140.8,144.9,147.1, 164.1；HRMS calcdfor C₆₈H₉₅ClN₆O₉S[M+H]⁺:1207.6643；found:1207.6628.

TC808:¹H NMR(800MHz,CDCl₃),(ppm)0.65 and 0.70(s,3H,CH₃),0.87-0.94(m,2H,CH₂), 0.96-1.63(m,30H,CH₃,CH₂and CH),1.73-2.02(m,10H,CH₂),2.19-2.77(m,10H,CH₂and CH), 3.40-3.52(m,4H,CH₂),3.55-3.73(m,12H,CH₂),3.81-3.89(m,5H,CH₂andCH),4.49-4.53(m, 4H,CH₂),4.65 and 4.67(s,2H,CH₂),4.82(br,1H,CH₂),5.31-5.33(m,1H,CH),6.85-7.14(m,2H, CH),7.44-7.52(m,5H,CH),7.66-7.88(m,4H,CH),8.17(br,1H,CH)；¹³C NMR(200MHz, CDCl₃),50.2,61.6,64.3,65.2,69.4,69.8,70.46,70.49,70.55,70.63,71.9,108.7,119.0, 122.7,123.9,125.4,126.5,127.5,129.2,137.2,147.1,164.2；HRMS calcd for C₆₇H₉₄ClN₇O₈S [M+2H]²⁺:596.8359；found:596.8366.

TC815:¹H NMR(800MHz,CDCl₃),(ppm)0.67(s,3H,CH₃),0.90(d,J＝6.4Hz,3H,CH₃), 0.91-0.98(m,2H,CH₂),0.99(s,3H,CH₃),1.01-1.58(m,30H,CH₂and CH),1.79-1.86(m,5H, CH₂),2.04-2.29(m,2H,CH₂),3.43(t,J＝7.2Hz,2H,CH₂),3.48-3.52(m,1H,CH₂),3.57-3.62(m, 7H,CH₂),3.65-3.68(m,4H,CH₂),3.72-3.74(m,2H,CH₂),3.84(t,J＝4.8Hz,2H,CH₂),3.88(t,J ＝4.8Hz,2H,CH₂),4.50-4.53(m,4H,CH₂),4.67(s,2H,CH₂),4.82(br,1H,CH),5.33-5.34(m, 1H,CH),6.84-7.13(m,2H,CH),7.44-7.50(m,5H,CH),7.66-7.88(m,3H,CH),8.17(br,1H,CH)；¹³C NMR(200MHz,CDCl₃),11.8,18.7,19.5,20.0,24.2,25.9,26.1,28.2,29.6,30.0,31.6, 31.85,31.86,35.7,35.8,36.4,37.2,39.7,42.3,50.1,50.2,56.0,56.7,64.5,65.2,69.4,69.6,69.8, 69.9,70.48,70.52,70.59,70.63,71.5,71.7,108.7,119.1,121.6,122.7,123.8,125.4,126.5,127.4, 129.2,137.2,140.8,144.9,147.1,164.2；HRMS calcd for C₆₈H₉₅ClN₆O₈S[M+2H]²⁺:596.3383； found:596.3377.

TC673:¹H NMR(800MHz,CDCl₃),(ppm)0.84-1.04(m,12H,CH₂and CH),1.17-2.02(m, 38H,CH₂and CH),2.20-2.54(m,9H,CH₂and CH),3.58-3.73(m,16H,CH₂),3,79(br,2H,CH₂), 3,87-3.88(m,2H,CH₂),4.22-4.25(m,1H,CH),4.40(br,2H,CH₂),4.52-4.57(m,3H,CH₂and CH),5.34-5.39(m,1H,CH),6.88-7.14(m,2H,CH),7.32-7.62(m,5H,CH),7.74-7.95(m,3H, CH),8.14-8.16(m,1H,CH)；¹³C NMR(200MHz,CDCl₃),29.7,31.3,41.5,70.3,70.5,70.7, 141.6,143.3,147.2,150.8,153.6,155.0,164.3；HRMS calcd for C₇₂H₉₆ClN₇O₁₁S[M+2H]²⁺: 651.8361；found:651.8367.

TC814:¹H NMR(800MHz,CDCl₃),(ppm)0.63(s,3H,CH₃),0.90(d,J＝6.4Hz,3H,CH₃), 0.92-0.98(m,1H,CH₂),1.01(s,3H,CH₃),1.06-1.64(m,24H,CH₂and CH),1.73-1.84(m,5H, CH₂),1.93-2.01(m,2H,CH₂),3.40(t,J＝7.2Hz,2H,CH₂),3.57-3.72(m,20H,CH₂),3.84(t,J＝ 4.8Hz,2H,CH₂),3.88(t,J＝4.8Hz,2H,CH₂),4.50-4.52(m,4H,CH₂),4.67(s,2H,CH₂),4.82(br, 1H,CH),6.84-7.13(m,2H,CH),7.44-7.49(m,5H,CH),7.65-7.87(m,3H,CH),8.17(br,1H,CH)；¹³C NMR(200MHz,CDCl₃),11.9,18.5,20.7,23.3,24.1,26.1,26.3,27.1,28.2,29.6,30.4, 31.6,31.9,34.5,35.3,35.5,35.8,36.4,40.1,40.3,42.0,42.6,50.1,56.0,56.4,64.5,69.4,69.6,69.7, 69.9,70.42,70.46,70.52,70.54,70.58,71.63,71.67,72.0,99.9,108.7,119.1,122.7,123.8,125.4, 126.5,127.4,129.2,137.1,144.8,147.1,163.6,164.2；HRMS calcd for C₆₇H₉₅ClN₆O₉S[M+2H]²⁺: 598.3358；found:598.3363.

TC829:¹H NMR(800MHz,CDCl₃),(ppm)0.84(s,3H,CH₃),0.85-0.98(m,1H,CH₂),0.99(s, 3H,CH₃),1.03-1.21(m,4H,CH₂and CH),1.25(s,3H,CH₃),1.38-1.75(m,12H,CH₂and CH), 1.81-1.83(m,2H,CH₂),1.94-1.97(m,2H,CH₂),2.07-2.09(m,1H,CH₂),2.14-2.29(m,4H,CH₂), 3.48-3.54(m,3H,CH₂and CH),3.59-3.63(m,6H,CH₂),3.65-3.68(m,4H,CH₂),3.72-3.73(m, 2H,CH₂),3.84(t,J＝4.8Hz,2H,CH₂),3.88(t,J＝4.8Hz,2H,CH₂),4.50-4.53(m,4H,CH₂),4.66 (s,2H,CH₂),4.81(br,1H,CH),5.32-5.33(m,1H,CH),6.85-7.13(m,3H,CH),7.44-7.50(m,5H, CH),7.65-7.88(m,4H,CH),8.17(br,1H,CH)；¹³C NMR(200MHz,CDCl₃),13.5,13.6, 19.3,20.0,20.9,22.4,23.7,25.6,31.3,31.6,31.7,36.4,37.2,40.1,41.9,42.2,42.6,50.0,50.2,56.8, 58.6,64.5,65.2,69.4,69.5,69.8,69.9,70.1,70.47,70.51,70.58,70.63,71.6,74.7,76.9,81.6, 108.7,119.2,121.4,122.7,123.8,126.6,127.4,137.2,140.8,144.8,147.1,163.7,164.3；HRMS calcd forC₆₈H₉₁ClN₆O₉S[M+2H]²⁺:602.3201；found:602.3198.

TC813:¹H NMR(800MHz,CDCl₃),(ppm)0.85(s,3H,CH₃),0.87-0.99(m,1H,CH₂),1.00(s, 3H,CH₃),1.04-1.21(m,4H,CH₂and CH),1.25(s,3H,CH₃),1.26-1.75(m,20H,CH₂and CH), 1.82-1.87(m,4H,CH₂),1.95-2.02(m,4H,CH₂),2.06-2.09(m,1H,CH₂),2.20-2.30(m,2H,CH₂), 2.38-2.48(m,4H,CH₂),3.42-3.52(m,3H,CH₂and CH),3.58-3.68(m,12H,CH₂),3.72-3.74(m, 2H,CH₂),3.86-3.88(m,4H,CH₂),4.51-4.53(m,4H,CH₂),4.68(s,2H,CH₂),4.81(br,1H,CH), 5.33-5.34(m,1H,CH),6.83(br,1H,CH),7.36-7.62(m,6H,CH),7.75-7.87(m,3H,CH), 8.26-8.36(m,1H,CH)；HRMS calcd for C₆₈H₉₅ClN₆O₉S[M+2H]²⁺:604.3358；found:604.3327.

TC807:¹H NMR(800MHz,CDCl₃),(ppm)0.86(s,3H,CH₃),0.87-0.99(m,2H,CH₂),1.00(s, 3H,CH₃),1.05-1.22(m,3H,CH₂and CH),1.26(s,3H,CH₃),1.26-1.77(m,20H,CH₂and CH), 1.83-1.85(m,2H,CH₂),1.95-2.02(m,4H,CH₂),2.07-2.09(m,2H,CH₂),2.16-2.30(m,4H,CH₂), 2.75 and 3.10(s,3H,CH₃),2.93-2.97(m,1H,CH),3.38-3.45(m,3H,CH₂and CH),3.50-3.54(m, 2H,CH₂),3.58-3.73(m,10H,CH₂),3.87-3.89(m,4H,CH₂),4.01and 4.05(s,3H,CH₃),4.17-4.24 (m,3H,CH₂),4.52-4.53(m,2H,CH₂),4.68 and 4.69(s,2H,CH₂),4.90-4.94(m,1H,CH), 5.34-5.35(m,1H,CH),6.58-6.61(m,1H,CH),7.12-7.13(m,1H,CH),7.23-7.26(m,1H,CH), 7.65-7.66(m,1H,CH),7.74(s,1H,CH),7.83(t,J＝8.0Hz,1H,CH),7.88(t,J＝8.0Hz,1H,CH), 8.05(q,J＝8.0Hz,1H,CH),8.12(d,J＝8.0Hz,1H,CH)；¹³C NMR(200MHz,CDCl₃), 13.6,19.4,20.9,22.3,23.7,24.2,26.0,26.4,29.3,29.5,30.1,31.3,31.6,31.7,36.5,37.2,40.1,42.2, 42.6,43.9,50.0,50.2,51.2,56.9,57.6,64.6,67.9,69.44,69.48,69.7,70.0,70.51,70.58,70.9,71.5, 71.7,75.2,121.5；HRMS calcd for C₆₆H₉₂F₃N₉O₉[M+2H]²⁺:606.8558；found:606.8561.

TC809:¹H NMR(800MHz,CDCl₃),(ppm)0.69 and 0.72(s,3H,CH₃),0.94-0.95(m,1H,CH₂), 1.00(s,3H,CH₃),1.03-1.63(m,26H,CH₂and CH),1.76-1.86(m,4H,CH₂),1.96-2.01(m,4H, CH₂),2.20-2.29(m,4H,CH₂),2.76 and 3.10(s,3H,CH₃),2.92-2.98(m,1H,CH),3.38-3.52(m, 5H,CH₂and CH),3.58-3.73(m,14H,CH₂),3.87-3.89(m,4H,CH₂),4.02and 4.06(s,3H,CH₃), 4.18-4.21(m,3H,CH₂),4.52-4.53(m,2H,CH₂),4.67 and 4.68(s,2H,CH₂),4.91-4.94(m,1H, CH),5.33-5.34(m,1H,CH),6.58-6.61(m,1H,CH),7.12-7.13(m,1H,CH),7.24-7.28(m,1H, CH),7.65-7.66(m,1H,CH),7.74-7.76(m,1H,CH),7.83(t,J＝8.0Hz,1H,CH),7.87(t,J＝8.0 Hz,1H,CH),8.03-8.06(m,1H,CH),8.12(d,J＝8.0Hz,1H,CH)；¹³C NMR(200MHz,CDCl₃), 12.1,12.4,,19.4,20.8,23.8,23.9,24.1,29.2,29.5,31.5,31.6,31.7,36.4,37.2,38.2,39.2, 42.0,42.2,49.87,49.94,50.2,51.2,56.5,64.5,67.9,69.41,69.44,69.6,69.9,70.48,70.50,70.55, 70.8,71.1,71.3,71.6,109.1,118.1,121.4,123.7,124.7,126.2,131.4,131.9,138.1；HRMS calcd for C₆₅H₉₁F₃N₁₀O₈[M+2H]²⁺:599.3559；found:599.3556.

TC836:¹H NMR(800MHz,CDCl₃),(ppm)0.67(s,3H,CH₃),0.87-0.88(m,1H,CH),0.89(d, J＝4.8Hz,3H,CH₃),0.91-0.99(m,5H,CH₂and CH),1.00(s,3H,CH₃),1.01-1.59(m,28H,CH₂ and CH),1.75-1.85(m,5H,CH₂),1.95-2.01(m,4H,CH₂),2.18-2.30(m,4H,CH₂),2.76and 3.10 (s,3H,CH₃),2.93-3.00(m,2H,CH),3.36-3.53(m,5H,CH₂and CH),3.58-3.73(m,12H,CH₂), 3.87-3.89(m,4H,CH₂),4.01 and 4.05(s,3H,CH₃),4.18-4.21(m,3H,CH₂),4.52-4.54(m,2H, CH₂),4.68 and 4.69(s,2H,CH₂),4.90-4.94(m,1H,CH),5.33-5.34(m,1H,CH),6.58-6.59(m,1H, CH),7.12-7.13(m,1H,CH),7.24-7.28(m,1H,CH),7.65-7.66(m,1H,CH),7.75(s,1H,CH),7.83 (t,J＝8.0Hz,1H,CH),7.88(t,J＝8.0Hz,1H,CH),8.05(q,J＝8.0Hz,1H,CH),8.12(d,J＝8.0 Hz,1H,CH)；¹³C NMR(200MHz,CDCl₃),13.6,19.4,20.9,22.3,23.7,24.2,26.0,26.4,29.3, 29.5,30.1,31.3,31.6,31.7,36.5,37.2,40.1,42.2,42.6,43.9,50.0,50.2,51.2,56.9,57.6,64.6,67.9, 69.44,69.48,69.7,70.0,70.51,70.58,70.9,71.5,71.7,75.2,121.5；HRMS calcd for C₆₆H₉₂F₃N₉O₈ [M+2H]²⁺:598.8583；found:598.8580.

TC838:¹H NMR(800MHz,CDCl₃),(ppm)0.84(s,3H,CH₃),0.86-0.99(m,2H,CH₂),1.00(s, 3H,CH₃),1.05-1.22(m,3H,CH₂and CH),1.26(s,3H,CH₃),1.27-1.75(m,16H,CH₂and CH), 1.82-1.85(m,3H,CH₂),1.93-2.02(m,4H,CH₂),2.08-2.28(m,8H,CH₂),2.40-2.43(m,2H,CH₂), 2.68 and 2.75(s,3H,CH₃),2.99 and 3.10(s,3H,CH₃),3.41-3.56(m,4H,CH₂and CH),3.61-3.73 (m,16H,CH₂),3.86-3.89(m,5H,CH₂),3.92(s,2H,CH₂),4.01and 4.05(s,3H,CH₃),4.18-4.21 (m,3H,CH₂),4.52-4.54(m,2H,CH₂),4.68 and 4.69(s,2H,CH₂),5.34-5.35(m,1H,CH), 6.55-6.60(m,1H,CH),7.11-7.14(m,1H,CH),7.20-7.27(m,1H,CH),7.64-7.67(m,1H,CH), 7.75-7.76(m,1H,CH),7.85-7.90(m,1H,CH),8.04-8.08(m,1H,CH),8.13(d,J＝8.0Hz,1H, CH),8.51-8.52(m,1H,CH)；¹³C NMR(200MHz,CDCl₃),13.5,13.6,19.3,20.0,20.9,22.4, 23.7,29.3,31.2,31.6,31.7,36.4,37.2,37.8,41.9,42.2,42.7,50.0,50.2,56.9,58.6,64.5,67.9,69.4, 69.6,69.9,70.1,70.5,70.8,108.7,126.3,132.1,134.0,138.4；HRMS calcd for C₆₆H₈₈F₃N₉O₉ [M+2H]²⁺:604.8401；found:604.8407.

TC821:¹H NMR(800MHz,CDCl₃),(ppm)0.84(s,3H,CH₃),0.86-0.98(m,1H,CH₂),1.00(s, 3H,CH₃),1.03-1.20(m,4H,CH₂and CH),1.24(s,3H,CH₃),1.26-1.82(m,22H,CH₂and CH), 1.95-1.97(m,1H,CH₂),2.06-2.08(m,1H,CH₂),2.21-2.26(m,2H,CH₂),3.43-3.58(m,8H,CH₂), 3.55-3.59(m,12H,CH₂),3.82-3.83(m,4H,CH₂),4.52-4.58(m,3H,CH),5.33-5.34(m,1H,CH), 7.50(br,1H,CH),7.78(br,1H,CH),7.85(br,1H,CH),7.94-7.95(m,1H,CH)；¹³C NMR(200 MHz,CDCl₃),13.6,19.4,20.9,22.3,23.7,24.2,26.0,26.3,29.5,30.0,31.2,31.5,31.7,36.4, 37.2,40.0,42.2,42.6,43.9,50.0,56.4,56.8,57.6,64.5,70.1,70.32,70.38,70.45,71.4,71.6,75.2, 121.5,126.8,140.7；HRMS calcd forC₆₂H₈₇Cl₂N₅O₁₂S[M+2H]²⁺:598.7797；found:598.7799.

TC822:Colorless oil,isolated yield 72％.¹H NMR(600MHz,CDCl₃),(ppm)0.67and 0.71(s, 3H,CH₃),0.86-0.94(m,2H,CH₂),0.98(s,3H,CH₃),1.04-1.68(m,26H,CH₂andCH),1.80-2.02 (m,8H,CH₂and CH),2.20-2.29(m,4H,CH₂),2.85-2.96(m,2H,CH₂),3.42-3.65(m,20H,CH₂), 3.80-3.85(m,4H,CH₂),4.49(t,J＝4.8Hz,2H,CH₂),4.60(s,1H,CH),5.31-5.32(m,1H,CH), 7.29-7.32(m,1H,CH),7.47(d,J＝9.0Hz,1H,CH),7.69(d,J＝6.5Hz,1H,CH),7.76-7.80(m, 3H,CH),7.84-7.96(m,4H,CH),8.62-8.63(m,1H,CH)；¹³C NMR(150MHz,CDCl₃),11.9, 12.6,14.1,15.4,16.2,19.34,19.38,20.7,20.8,22.6,23.6,24.3,25.7,26.8,27.2,28.7,29.3,29.7, 31.5,31.6,36.4,36.5,37.2,39.0,42.1,42.3,42.4,42.5,49.7,50.1,52.2,56.2,56.4,56.7,64.2,64.5, 69.3,69.6,69.9,70.1,70.3,70.39,70.42,70.67,70.71,71.5,121.2,121.8,122.8,123.1,124.1, 125.2,126.7,127.5,129.7,130.0,130.6,132.0,136.2,137.1,139.1,140.4,140.7；HRMS calcd forC₆₁H₈₆Cl₂N₆O₁₁S[M+2H]²⁺:591.2799；found:591.2798.

TC823:¹H NMR(800MHz,CDCl₃),(ppm)0.67(s,3H,CH₃),0.90(d,J＝6.4Hz,3H,CH₃), 0.91-0.97(m,1H,CH₂),0.99(s,3H,CH₃),1.04-1.57(m,22H,CH₂and CH),1.80-1.84(m,2H, CH₂),1.95-2.00(m,2H,CH₂),2.21-2.27(m,2H,CH₂),3.44-3.60(m,20H,CH₂),3.83-3.90(m,4H, CH₂),4.57-4.59(m,3H,CH),5.33-5.34(m,1H,CH),7.53(br,1H,CH),7.85(br,2H,CH),7.95(br, 1H,CH),8.10(br,2H,CH)；¹³C NMR(200MHz,CDCl₃),11.8,18.6,19.3,21.0,24.2,25.9, 26.1,28.2,29.6,29.9,31.6,31.8,35.6,35.8,36.4,37.2,39.7,42.19,42.24,50.0,56.0,56.4,56.7, 64.5,69.9,70.11,70.33,70.38,70.47,71.5,71.7,75.2,121.6,126.8,129.7,132.7,140.3,140.7, 142.5,143.6；HRMS calcd for C₆₂H₈₇Cl₂N₅O₁₁S[M+2H]²⁺:590.7823；found:590.7816.

TC839:Colorless oil,isolated yield 45％.¹H NMR(600MHz,CDCl₃),(ppm)0.83(s,3H,CH₃), 0.87-0.93(m,1H,CH₂),0.99(s,3H,CH₃),1.02-1.19(m,3H,CH₂and CH),1.24(s,3H,CH₃), 1.26-2.42(m,26H,CH₂and CH),3.43-3.84(m,24H,CH₂),4.50-4.60(m,3H,CH),5.30-5.33(m, 1H,CH),7.47(br,2H,CH),7.75-7.95(m,6H,CH)；¹³C NMR(150MHz,CDCl₃),13.5,13.8, 19.3,20.3,20.8,22.4,23.7,25.4,31.2,31.5,31.7,36.4,37.1,40.0,41.9,42.1,42.6,49.9,56.4,56.8, 58.5,64.5,69.1,69.6,69.7,69.99,70.04,70.2,70.3,70.4,71.6,74.7,77.5,82.2,121.4,121.8, 126.8,129.7,131.9,140.4,140.7,142.5,164.0；HRMS calcd for C₆₂H₈₃Cl₂N₅O₁₂S[M+2H]²⁺: 596.7641；found:596.7639.

TC1055:¹H NMR(800MHz,CDCl₃),(ppm)0.84(s,3H,CH₃),0.87-0.92(m,1H,CH₂),1.00(s, 3H,CH₃),1.05-1.20(m,4H,CH₂and CH),1.23(s,3H,CH₃),1.26-2.25(m,24H,CH₂and CH), 3.40-3.64(m,10H,CH₂),3.73-3.84(m,4H,CH₂),5.30-5.34(m,1H,CH),7.31-7.44(m,2H,CH), 7.71-7.94(m,3H,CH)；¹³C NMR(200MHz,CDCl₃),13.6,19.4,20.9,22.4,23.7,24.1,26.0, 26.3,29.5,30.0,31.3,31.6,31.7,36.4,37.2,40.0,42.2,42.6,43.8,50.0,56.6,56.8,57.6,64.6,70.0, 70.3,71.7,75.2,121.5,140.7；HRMS calcd forC₅₆H₇₅Cl₂N₅O₉S[M+2H]²⁺:532.7404；found: 532.7390.

TC1056:¹H NMR(500MHz,CDCl₃),(ppm)0.84(s,3H,CH₃),0.87-0.92(m,1H,CH₂),0.99(s, 3H,CH₃),1.05-1.20(m,4H,CH₂and CH),1.25(s,3H,CH₃),1.26-2.25(m,24H,CH₂and CH), 3.39-3.72(m,14H,CH₂),3.81-3.90(m,4H,CH₂),5.30-5.34(m,1H,CH),7.46-7.66(m,2H,CH), 7.82-7.96(m,3H,CH)；¹³C NMR(125MHz,CDCl₃),13.6,19.3,20.8,22.2,23.6,24.1,25.9, 26.2,29.4,30.0,31.2,31.5,31.6,36.4,37.1,40.0,42.1,42.5,43.8,49.9,56.4,56.8,57.5,64.4,70.0, 70.27,70.35,70.43,71.4,71.5,75.1,109.9,121.4,126.8,140.7；HRMS calcd for C₅₈H₇₉Cl₂N₅O₁₀S [M+2H]²⁺:554.7535；found:554.7539.

TC1057:¹H NMR(800MHz,CDCl₃),(ppm)0.84(s,3H,CH₃),0.87-0.92(m,1H,CH₂),1.00(s, 3H,CH₃),1.04-1.21(m,3H,CH₂and CH),1.25(s,3H,CH₃),1.31-1.84(m,16H,CH₂and CH), 1.96-2.08(m,2H,CH₂),2.20-2.29(m,2H,CH₂),3.36-3.66(m,20H,CH₂),3.82-3.87(m,4H,CH₂), 5.32-5.34(m,1H,CH),7.33-7.47(m,2H,CH),7.63-7.96(m,6H,CH)；¹³CNMR(200MHz, CDCl₃),13.6,19.3,20.9,22.3,23.7,24.1,26.0,26.3,29.4,29.5,30.0,31.2,31.6,31.7,36.4, 37.2,40.0,42.2,42.6,43.8,50.0,56.4,56.8,57.6,64.4,69.1,69.3,69.66,69.71,69.81,69.89, 70.13,70.36,70.40,70.45,71.42,71.65,75.16,121.5,126.9,129.7,131.8,140.8；HRMS calcd for C₆₀H₈₃Cl₂N₅O₁₁S[M+2H]²⁺:576.7666；found:576.7652.

实施例3

分别对实施例2中合成的各配体进行细胞活性实验：

1、细胞活性实验：使用NIH 3T3细胞进行荧光素酶报告实验测定配体化合物作为SMO 受体拮抗剂的活性。信号使用100nM SAG(Sellect，货号S7779)激活。具体步骤如下：转基因有荧光素酶报告基因NIH3T3的细胞(Clontech，货号631197)在96孔板中培养至(6*10⁵) 个细胞/孔，然后加入浓度为10000nM、2000nM、400nM、80nM、16nM、3.2nM、0.64 nM、0.128nM、0.0256nM、0.00512nM的梯度浓度的配体化合物在37℃共孵育1小时，加入终浓度为100nM的激活剂SAG，继续在37℃共孵育24小时。加入

荧光素报告系统(Promega，货号，货号E2920)并使用Envision酶标仪(PerkinElmer)对Gli荧光素酶报告水平进行测定。平行测定三组数据，根据下述公式拟合曲线并得到化合物的IC50值。

Y＝Bottom+(Top-Bottom)/(1+10^((LogIC50-X)*HillSlope))

其中，X代表化合物浓度，Y代表酶标仪测定的荧光读数，Top表示曲线的上平台值，Bottom代表曲线的下平台值。HillSlope代表曲线最大斜率的绝对值。

2、实验结果如表2所示：

表2

上表中的部分双头化合物在细胞活性实验中的EC₅₀值优于相应的母体分子，说明这些化合物能够通过同时结合平滑受体的胞外域和跨膜域的方式来增强其结合力以及细胞活性，因此这一策略可以应用于合成高活性的靶向平滑受体的小分子药物。

实施例4

已报道的晶体结构中，胞外域配体一般为甾体，跨膜域配体结构则更类似于一般的合成药物分子(图1B)。为了进一步研究受体共结晶的小分子配体的结合取向以及两个结合域之间距离等信息(图1A)，并为了确认连接体在双头配体中的作用，可以通过一系列连接体长度不同，而两端用于结合平滑受体的药效团相同的双头分子，考察其活性与连接体长度的关系。具体的，将其中连接体长度逐渐变化的OHC-SAG型双头分子的连接体长度及活性信息归纳如下表所示：

表3

连接体长度与活性之间的关系如图3所示。由图3可知，当连接体长度合适时，双头配体的活性可达到最优。其原理可由图2解释：当连接体长度短于结合位点之间的距离时，同一个双头分子的两个药效团无法同时结合SMO受体的两个位点，因此无法产生协同效应并增强活性(图2a)；若连接体长度与结合位点间距相符，在一定长度范围内，两个药效团可以同时结合SMO受体的两个位点，由于协同效应，其结合贡献相互增强，表现为活性增强(图2b)；若连接体长度进一步增加，由于熵效应导致协同效应消失，两个药效团无法有效增强活性，因此活性有所下降(图2c)。

综上所述，本发明有效克服了现有技术中的种种缺点而具高度产业利用价值。

上述实施例仅例示性说明本发明的原理及其功效，而非用于限制本发明。任何熟悉此技术的人士皆可在不违背本发明的精神及范畴下，对上述实施例进行修饰或改变。因此，举凡所属技术领域中具有通常知识者在未脱离本发明所揭示的精神与技术思想下所完成的一切等效修饰或改变，仍应由本发明的权利要求所涵盖。

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