作者要感谢JUMPEI森本博士和托德多伦博士宝贵的援助。这项工作是由来自NHLBI（NO1-HV-00242）的合同支持。

1，斯普利特和池合成基础知识为了有效地产生在固相上的大量的化合物，分裂和池合成经常被用作一个一般的策略。 如图4的TentaGel珠是第一分割成三个部分。各部分进行反应以不同的试剂，生成的小珠的第一个残基。在第一反应后，所有的三个部分被汇集在一起​​，混合，然后再分成三个部分。每个部分将再次用不同的试剂反应，生成对珠子的第二个残基。经过两次分裂和池步骤，生成九种化合物。 在子单体合成，珠粒首先分成几个部分，以与不同溴代羧酸在偶联试剂的存在下进行反应。用溶剂洗涤后，所有的珠将被合并在一起并混合，然后再分成几个部分，以与不同的反应伯胺。胺化后，所有珠子被汇集在一起​​，并彻底清洗，在完成每个珠子上一个完整的单体。这个过程可以重复，直到期望的分集为止。 2，准备溴化酸从天然氨基酸在子单体合成，每种单体的合成中被分成两个独立的步骤：（1）耦合酰溴和2胺化伯胺（ 图2）。为了合成肽叔酰胺，手性酸溴化物与在α碳侧链将由天然氨基酸制备。这里我们描述的转化天然氨基酸成相应的酸溴化物，高立体声保真度的方法。我们用丙氨酸作为一个例子;其它氨基酸包括丝氨酸，苏氨酸，天冬氨酸，谷氨酸，天冬酰胺，谷氨酰胺，甘氨酸，缬氨酸，异亮氨酸，苯丙氨酸，也可以类似的天气下下转化成溴酸纳秒。请注意，某些氨基酸与官能团如酚，胍胺和需要进行变换之前受到保护。反应设置示于图3。 安全注意事项：对于需要涉及溴化氢，亚硝酸钠等腐蚀性/有毒化学物质，如护目镜，实验室外套，和耐化学腐蚀手套适当的安全设备下列反应。所有的反应应在通风橱中由有经验的化学家进行。 <ol><li>要加370毫升水中成630毫升48％的HBr溶液，以制备1升，30％的HBr溶液。加入500毫升乙二醇到1L浴容器;添加干冰以保持温度在-10℃。注意：48％的HBr水溶液呈强酸性，腐蚀性，小心处理。请认真阅读MSDS方可使用。 </li><li>添加D-丙氨酸（8.9克，0.1摩尔）和溴化钾（11.9克，0.1摩尔），以250毫升的三颈圆底烧瓶中磁力搅拌棒。加入100毫升，并在T准备了30％的HBr他前面的步骤。把该烧瓶中，在步骤2.1中制备的乙二醇浴中，并保持温度在-10℃。通过从烧瓶中10分钟的底部的长针， 如图3泡氩气中搅拌，以300rpm的磁力搅拌棒的溶液。 </li><li>在一个100毫升烧杯中，用20ml水溶解的NaNO 2（8.28克，0.12摩尔）。添加到溶液中的压力平衡滴液漏斗和密封滴液漏斗用隔膜。慢慢打开滴液漏斗中的阀并让的NaNO 2溶液滴入烧瓶中。控制阀门以调节滴落速率每秒约2滴。保持搅拌在300rpm，保持氩气从烧瓶底部鼓泡。烧瓶中应保持在乙二醇浴中于-10℃，直到所有的NaNO 2溶液。注意：该步骤中加入的NaNO 2溶液中产生热量和气体。滴水率应仔细CON受控和整个系统应通过氩气口是开放的。 </li><li>保持搅拌3更小时，并让温度从-10℃升温至室温。将得到的溶液应该是无色至淡黄色;如果颜色太暗，施加真空以除去过量的氮氧化物和可能的溴2在反应过程中生成的。 </li><li>从该溶液中使用的提取漏斗3倍35毫升乙醚提取产物。合并有机相并用饱和食盐水洗涤。将有机相也可与少量的NaHCO 3的前，用盐水洗涤以除去颜色，如果是暗洗涤。干燥有机相经Na 2 SO 4 6小时。 </li><li>过滤出用Na 2 SO 4和蒸发在真空下将溶剂，粗产物应得到明确至浅黄色油状物。粗产物可以通过蒸馏进一步在115℃下进行纯化，3个毫米汞柱，或由硅胶柱上用3:1己烷：乙酸乙酯。 </li><li>纯的产物，得到为透明油状物，6.6克（收率74％），密度=1.69克/ ml时，[α] D20 = 24°（甲醇），1 H NMR（400兆赫，CDCL 3）δ4.41（Q， J = 7.0赫兹，1H），1.86（D，J = 7.0赫兹，3H）。中的（S）-2 - bromopropanoic的情况下酸-D 4（从D 4-L-丙氨酸的方法制备），纯的产物，得到为透明油状物，产率78％，密度= 1.87克/毫升，[α] D20 = -19°（甲醇）1 H-NMR，无显著H信号被观察到。 ESI-MS - [M-1] - = 155.1（预期154.97）。为（S）-2 -溴-4 -甲基戊酸（从L-亮氨酸使用相同的程序制备的）的纯产物，得到为透明油状物，产率89％，[α] D20 = 37°（甲醇），1H NMR（400兆赫，CDCL 3）δ4.30（T，J = 7.7赫兹，1H），1.94（日，J = 10.8，3.9赫兹，2H），1.81（TT，J = 13.2，6.5赫兹，1H），0.96 （日，J = 18.2，6.6赫兹，7H）。中的（S）-2 - 溴-3 - 苯丙氨酸的情况下纯的产物，得到为淡黄色油nylpropanoic酸（由L-苯丙氨酸用同样的步骤制备），产率72％，[α] D20 = 17°（甲醇），1 H NMR（400兆赫，CDCL 3）δ7.38 - 7.19（M，5H），4.42（日，J = 8.1，7.3赫兹，1H），3.47（日，J = 14.2，8.2赫兹，1H），3.25（日，J = 14.2，7.2赫兹，1H）。 </li></ol>丙氨酸3，使用转氨酶同位素标记在组合文库的合成，尤其是在一珠一化合物（OBOC）库的分裂 - 和 - 池的合成，化合物，可以从每个胎圈获得的量是比较小的。 （通常1皮摩尔至10纳摩尔）。此外，质谱法被广泛地用于最终化合物，由于其高灵敏度的鉴定和表征。为了使用质谱分析，以确定在最终的PTA产物的手性中心的绝对构型，溴代酸对映体应isoto使用前pically标记。在这里，我们将介绍使用转氨酶和D 2 O来标记L-丙氨酸的方法。 <ol><li>溶解L-丙氨酸（300毫克，3.36毫摩尔）在50毫升聚乙烯管10毫升D 2 O中的。添加α-酮戊二酸（10毫克，0.068毫摩尔）作为共底物。热身管到37℃，用1M NaOD soution调整的PD 8.5-8.7。注：PD受pH值试纸测定。传统的电子pH计配有玻璃电极选择性的H +可以提供不正确的读出D +。 </li><li> （从猪心脏，罗氏诊断，印第安纳波利斯，在0.1毫克，EC 2.6.1.2）添加谷丙转氨酶的PD 8.5 - 8.7，从先前的步骤中制备37℃的解决方案。把该管在37℃的培养箱中，轻微摇动过夜孵育它，10至30rpm的转速是优选的。 </li><li>过夜孵育后，取0.5毫升D 2 O中的溶液中，并用1 H-NMR检查反应进程。的所有质子信号lanine，δ3.76（Q，J = 7.2赫兹，1H），1.46（D，J = 7.3赫兹，3H），1 H NMR 400 MHz的，应大大由于氘化抑制。如先前描述的23的98％以上的质子应该交换到氘。注：D 2 O可通过蒸馏部分恢复，如果大规模进行反应（&gt;200毫升D 2 O）。通常情况下，60％到80％D 2 O中的可以从溶液中蒸去溶剂。 </li><li>冷冻用液氮将上述溶液，并用冷冻干燥机，以获得白色氘化L-丙氨酸粉末冻干它。 </li></ol>拟肽接头区的4合成不需要PTA文库合成的接头区。然而，为了避免高背景中的MALDI质谱的低分子量范围（100-600），并改善该化合物的电离中，拟肽连接子具有多个极性残基是常用的。这种拟肽林KER可以通过标准的拟肽合成方法来合成。在这里我们将合成的N-甲氧基乙基甘氨酸一个五聚体作为连接体（ 如图5）。 <ol><li>在10毫升DMF中溶胀90μm的珠粒的TentaGel带RAM接头（1克，0.27毫摩尔/克）3小时，在12毫升注射器的反应器，轻微摇动。 </li><li>从反应器排出的二甲基甲酰胺，并加入10 ml 20％哌啶的DMF溶液从Rink酰胺连接器脱保护Fmoc基团。摇珠30分钟20％哌啶的解决方案。用DMF洗涤5倍，除去所有的哌啶。 </li><li>取数珠从注射器，并与四氯苯醌试验测试。珠应该把深褐色（四氯苯醌试验阳性伯胺），如果FMOC成功脱保护。 </li><li>准备了以下解决方案：1 20毫升，2M的溴乙酸/ DMF溶液; 2 20毫升，2M的DIC / DMF溶液; 3，10毫升，1M的methoxylethylamine / DMF溶液。 </li><li>加入5毫升2M的溴乙酸/ DMF溶液，以珠，轻轻摇晃。然后加入5毫升2M的DIC / DMF溶液于珠子;密封注射器的活塞，并把它放在摇床。摇动10分钟。 </li><li>彻底冲洗用DMF的珠子。加入2毫升1M甲氧基乙胺/ DMF溶液从步骤4.4制得的珠子。密封注射器的柱塞和振动筛30分钟摇晃。 </li><li>用DMF洗涤5倍的珠子。检查数珠与四氯苯醌试验，若阳性（珠变成蓝色），然后继续进行下一个步骤。否则，重复步骤4.6。 </li><li>重复步骤4.5至4.7，4倍，完成五聚体。 </li></ol> 5，分体式和池PTA图书馆与（R）的合成 - 和（S）-2 - 溴酸在这里，我们描述了与使用1克珠从步骤4.8 9,261化合物的理论多样性的小PTA库的合成。需要注意的是一个90微米的TentaGel珠每克含约2,900,000珠;因此，该冗余该库将是2.9×10 6 / 9,261 = 312份。我们将用溴乙酸，（R）-2 - bromopropanoic和同位素标记的（S）-2 -溴丙酸-D 4为羧酸，和7不同的胺（A1〜A7，参见图5为详细说明）进行胺化。注射器的反应器和真空歧管将被用来执行合成。 <ol><li>加入10 mL 1:1 DCM：DMF从步骤4.8注射器;使用1000微升移液器带有截断枪头平摊所有1 g树脂分为三个5毫升注射器反应堆。将它们标记为B（溴乙酸）中，R（（R）-2 - bromopropanoic）和S（（S）-2 -溴丙酸-D 4）。与DCM的3倍洗净所有3注射器和清洗注射器标有R和S采用无水THF 3倍，洗注射器标记为B用DMF 3倍。 </li><li>注射器R和 S。 BTC耦合溴丙酸的。 <ol><li>准备一个新鲜的BTC / THF溶液。添加pproximately 200毫克的BTC到小瓶在通风橱中，用盖子密封。称重在小瓶BTC的量。计算所需的溶剂量，并加入无水THF放入小瓶中，使20毫克/毫升的BTC / THF溶液。 </li><li>准备溴酸/ BTC的混合物。加（R）-2 -溴丙酸（89微升，0.95毫摩尔）和（S）-2 -溴丙酸-D 4（89微升，0.95毫摩尔）在2小药瓶分开。向每个小瓶中，加入5毫升上述20毫克/毫升的BTC / THF溶液中。密封两个小瓶，把他们在-20℃下冷冻20分钟。 </li><li>新增1,125微升，2:1 THF / DIPEA（750微升焦磷酸钾，375微升DIPEA，2.2毫摩尔）分别注射器R和 S。混合珠枪头。让他们坐了5分钟。 </li><li>取上述两种冷却溴酸/ BTC混合物从步骤5.2.2，添加2,4,6 - 三甲基吡啶（356微升，2.7毫摩尔），以每小瓶。白色的沉淀物会立即形成。直接套用相应的悬挂在碱化珠（步骤5.2.3注射器R和S），尽快，然后把它们放在摇床下120转摇2小时。 注意：在注射器反应器中的溶液应在反应的整个过程中，淡黄色悬浮液。较深的颜色是过热的迹象初始加酸氯化物溶液中释放出来。这可以通过进一步冷却得到解决或稀释的溴酸/ BTC的混合物。 </li></ol></li><li>注射器乙 。溴乙酸耦合DIC <ol><li>准备一个新的20毫升，2M的溴乙酸/ DMF溶液。制备一个20毫升，2M的DIC / DMF溶液。 </li><li>加入2毫升2M的溴乙酸/ DMF溶液注射器，B，轻轻摇晃。加入2毫升2M的DIC / DMF溶液注射器，B，轻轻摇晃。 </li><li>放注射器B上的相同摇床如注射器R和 S </strong&gt;，摇动2小时。需要注意的是溴乙酸/ DIC的偶联反应在30分钟内完成;延长反应时间为分裂和池合成的便利。 </li></ol></li><li> 2小时后，取注射器R，S和B的振动筛。与DCM的5倍，彻底冲洗所有三个注射器。然后用DMF 5倍。需要注意的是注射器R和S不 ​​能用DMF洗涤被用DCM或THF第一前洗涤。 </li><li>池从注射器R，S和B中的所有微珠成一个12毫升注射器反应器中。用DMF 5X清洗所有的珠子。 </li><li>加入10 ml 1:1 DCM：DMF中的注射器;使用1000微升移液器带有截断枪头平分所有的珠串成7个人2毫升注射器，将它们标记为A1-A7。 </li><li>胺化。制备10毫升，2M的伯胺/ DMF溶液为每个在图5中列出的7胺。加入5 ml EAC的ħ胺溶液到相应的注射器A1-A7。所有孵育7注射器在60℃培养箱中振荡培养过夜。 </li><li>孵育后，用DMF彻底清洗所有的珠子。花几分钟的珠子从每个注射器，并检查与四氯苯醌的测试。如果珠变绿（正）在3分钟内，继续进行下一个步骤。如果是负数，为负注射器重复步骤5.7。 </li><li>重复步骤5.1至5.8倍，完成三聚体。可选步骤：在每个周期结束后，我们建议通过质谱法如下所述检查综合素质。所有9,261化合物现在对合成的TentaGel珠OBOC库。 </li><li>质谱家教会的确认。 家教会是高度结构化的低聚物和具有N-甲基化肽的许多共同特点。之一的N-甲基化的肽的固相合成中的常见问题是在TFA裂解的酸降解。为了抑制酸降解，如环孢霉素分子的裂解从固体支持物通常在低温下进行。我们比较了用于从固体支持物上切割不同的PTA分子的各种裂解的条件。我们发现，一般来说，温度低，并减少TFA的浓度可有效抑制酸的降解，并提供更纯的化合物。 <ol><li>制备出10毫升15毫升管1时01 TFA / DCM溶液中。密封管，并把它在-20℃下冷冻20分钟。 </li><li>洗需要用DCM 5倍被切割的小珠。摇在DCM珠子15分钟，并用DCM 5倍再次洗涤磁珠。 </li><li>从注射器排出的DCM。使用光镜和一个吸管截断尖到每孔各个别珠转移到96孔板中，一个胎圈。 </li><li>覆盖96孔板盖玻片。把板在-20℃下冷冻15分钟。 </li><li>采取冷却1时01分TFA / DCM解决方案从步骤5.10.1，并添加20μl到每个包含一个珠孔中。把盖玻片背部和普吨的96孔板上在-20°C冰箱摇床。 </li><li>摇动20分钟。就拿96孔板出来，撕下盖玻片。吹干通过吹入空气或氩气在它从每个孔中的TFA / DCM。如果超过10个珠被裂解，一个Speed​​Vac中可以用来从整个板干燥TFA / DCM。请注意，在这一点上，不是所有的化合物的裂解掉珠，但在切割的化合物应该是绰绰有余进行质谱分析。 </li><li>加20微升6点04的ACN：H 2 O溶液，以每孔溶解在切割的化合物。现货0.6微升每种化合物溶液与对MALDI板0.6微升CHCA MALDI基质在一起。 </li><li>用MALDI质谱分析，以确定每个化合物的分子量和序列（MS / MS）。 </li></ol></li></ol> 6，四氯苯醌测试<ol><li>准备以下试剂新鲜为每个测试。溶液A：2％四氯苯醌（CAS：118-75-2）的DMF。解B：2％乙醛（CAS：75-07-0）的DMF。 </li><li>混合100微升溶液A用100μl溶液B的试验在1.5ml管之前;放下珠，轻轻摇晃。如果小珠变成5分钟内的蓝色，它表示的仲胺的小珠的表面上的存在。伯胺得到深棕色的颜色，而不是脸色发青。 </li></ol>

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作者宣称，他们有没有竞争的财务权益。

肽叔酰胺（自由贸易协定）是肽低聚物的超家族。除了充分研究的肽，拟肽和N-甲基化肽，这个家庭中的化合物有很大一部分仍然是充分研究，majorly由于缺乏接入通用N-烷基化肽的合成方法。这里我们描述了一种有效的方法来合成贸易协定与从氨基酸衍生的手性结构单元。此前，我们曾报道使用一个新的子单体路线的PTA分子23的合成库。我们已经表明，自由贸易协定是其通过骨干具有构象?...

肽模拟物是模拟天然肽的结构的化合物。它们被设计为保留的生物活性，同时克服了与天然肽，包括细胞的渗透性和稳定性对蛋白水解1-3有关的问题。由于这些化合物的低聚物的性质，大合成文库可以容易地通过单体或子单体的合成路线4-7访问。一个模拟肽的研究最多的类别是拟肽。拟肽是，可以很容易地使用子单体策略8,9来合成N-烷基化甘氨酸的低聚物。许多有用的蛋白质配体已成功识别从筛选对蛋白指标1，10-14大合成拟肽库。不过，从拟肽库确定了“命中”很少归档非常高的亲和力对蛋白质靶1,10-14,22。一马约旦拟肽和天然肽之间的区别是，大部分的拟肽的普遍缺乏，以形成二级结构，由于缺乏手性中心和构象约束的能力。为了解决这一问题，开发了在过去的十年多种策略，主要着眼于包含在主链氮原子的15-22侧链的修饰。最近，我们已经开发出一种新的合成路线，以引进天然氨基酸侧链到拟肽骨干创造肽叔酰胺23。 肽叔酰胺（贸易协定）的肽模拟物，包括但不限于肽（R 2 = H），拟肽（R 1 = H）和N-甲基化的肽的超家族（R 1≠H，R 2 = Me）的。 （ 见图1）本合成路线采用天然存在的氨基酸为手性和侧链上的源45; - 碳，和市售的伯胺，得到N-取代。因此，比单纯肽，拟肽或N-甲基化多肽的化学较大的空间可以探索。圆二色光谱表明，PTA分子是高度结构化的解决方案。在PTA-蛋白质复合物的一个表征清楚地表明，PTA的构象限制所需要的具有约束力。最近，我们还发现，一些PTA分子具有改善细胞通透性比他们的拟肽和肽同行。我们相信，这些PTA库可以是高亲和力配体蛋白指标的良好来源。在本文中，我们将讨论的样本中以及这些化合物的耦合和裂解一些改善条件细节一珠一化合物（OBOC）PTA库的合成。

<table border="0" cellpadding="0" cellspacing="0" width="889">
	<tbody>
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			<td height="21" style="height:21px;width:249px;">Acetaldehyde</td>
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			<td height="21" style="height:21px;width:249px;">Acetonitrile</td>
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			<td height="21" style="height:21px;width:249px;">Anhydrous Tetrahydrofuran (THF)</td>
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			<td>CAS:109-99-9</td>
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			<td height="21" style="height:21px;width:249px;">Benzylamine</td>
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			<td height="21" style="height:21px;width:249px;">bis(trichloromethyl) carbonate (BTC)</td>
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			<td height="21" style="height:21px;width:249px;">Bromoacetic acid</td>
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			<td height="21" style="height:21px;width:249px;">Chloranil</td>
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			<td height="20" style="height:21px;">D2O</td>
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			<td style="width:160px;">BJ-NS300-20</td>
			<td>CAS:75-09-2</td>
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			<td style="width:160px;">BJ-076-4</td>
			<td>CAS:68-12-2</td>
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			<td height="20" style="height:21px;width:249px;">Isopentylamine</td>
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			<td height="20" style="height:21px;width:249px;">KBr</td>
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			<td style="width:160px;">424070025</td>
			<td>CAS:7758-02-3</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">L-alanine</td>
			<td style="width:169px;">Anaspec</td>
			<td style="width:160px;">61385-100</td>
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			<td height="20" style="height:21px;width:249px;">3-Methoxypropylamine</td>
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			<td height="20" style="height:21px;width:249px;">2-Methoxyethylamine</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">143693</td>
			<td>CAS:109-85-3</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">N-(3-Aminopropyl)-2-pyrrolidinone</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">136565&#160;</td>
			<td>CAS:7663-77-6&#160;</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">N,N&#39;-Diisopropylcarbodiimide (DIC)</td>
			<td style="width:169px;">ACROS</td>
			<td style="width:160px;">115211000</td>
			<td>CAS:693-13-0</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">N,N-Diisopropylethylamine (DIPEA)</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">D125806</td>
			<td>CAS:7087-68-5</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;">NaNO2</td>
			<td style="width:169px;">ACROS</td>
			<td style="width:160px;">424340010</td>
			<td>CAS:7631-99-4</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:21px;width:249px;">NAOD 40% solution in water</td>
			<td style="width:169px;">ACROS</td>
			<td style="width:160px;">200058-506</td>
			<td>CAS:7732-18-5</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Piperidine</td>
			<td style="width:169px;">ALFA AESAR</td>
			<td style="width:160px;">A12442-AE</td>
			<td>CAS:110-89-4</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Piperonylamine</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">P49503&#160;</td>
			<td>CAS:2620-50-0</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Propylamine</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">240958</td>
			<td>CAS:107-10-8</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Trifluoroacetic acid</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">299537</td>
			<td>CAS:76-05-1</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">&#945;-Cyano-4-hydroxycinnamic acid&#160;</td>
			<td style="width:169px;">Sigma-Aldrich</td>
			<td style="width:160px;">39468</td>
			<td>CAS:28166-41-8&#160;&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">&#945;-ketoglutarate</td>
			<td style="width:169px;">ALFA AESAR</td>
			<td style="width:160px;">AAA10256-22</td>
			<td style="width:311px;">CAS:328-50-7</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Tentagel Resin with RINK linker</td>
			<td style="width:169px;">Rapp-Polymere</td>
			<td style="width:160px;">S30023</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Alanine transaminase</td>
			<td style="width:169px;">Roche</td>
			<td style="width:160px;">10105589001</td>
			<td>AKA: Glutamate-Pyruvate Transaminase (GPT)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:249px;">Incubator</td>
			<td style="width:169px;">New Brunswick Scientific</td>
			<td style="width:160px;">Innova44</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">NMR</td>
			<td style="width:169px;">Bruker</td>
			<td style="width:160px;">400MHz</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">MALDI mass spectrometer</td>
			<td style="width:169px;">Applied Biosystems&#160;</td>
			<td style="width:160px;">4800 MALDI-TOF/TOF</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Lyophilizer</td>
			<td style="width:169px;">SP Scientific</td>
			<td style="width:160px;">VirTis benchtop K</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Syringe reactor</td>
			<td style="width:169px;">INTAVIS&#160;</td>
			<td style="width:160px;">Reaction Column</td>
			<td>3ml, 5ml, 10ml, 20ml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:249px;">Vacuum manifold&#160;</td>
			<td style="width:169px;">Promega</td>
			<td style="width:160px;">A7231</td>
			<td>Vac-Man</td>
		</tr>
	</tbody>
</table>

split-and-pool synthesis and characterization of peptide tertiary amide library

肽是蛋白质配体的巨大来源。这些化合物的低聚性质使我们利用组合化学访问的固相合成大库。一个模拟肽的研究最多也类是拟肽。拟肽是易于合成，并已被证明是蛋白酶解抗性和细胞渗透性。在过去的十年中，许多有用的蛋白质配体已通过拟肽库的筛选鉴定。然而，大多数从拟肽库确定的配体不显示高亲和力，除了极少数例外。这可能是由于，在某种程度上，缺乏手性中心的构象和限制在类肽分子。最近，我们描述了一种新的合成路线来访问肽叔酰胺（家教）。贸易协定是肽模拟物，包括但不限于肽，拟肽和N-甲基化的肽的一个家族。与在两个α-碳和主链氮原子的侧链这些分子的构象可得到很大的空间位阻和烯丙基1,3应变的限制。 （ 图1）我们的研究表明，这些PTA分子是高度结构化的溶液中，并且可以被用来确定蛋白质的配体。我们相信，这些分子可以是高亲和力的蛋白配体未来的源泉。在这里，我们描述的合成方法结合两种分裂和池和子单体策略的力量来合成样品家教会的一珠一化合物（OBOC）库。

在这里，我们将展示从PTA三聚体与接头三个有代表性的电离光谱。 如图6A所示 ，当用50％TFA / DCM溶液在室温下裂解，显著降解是观察。在图6A中 ，峰593和484分别对应于连接体和三聚体的PTA，表明整个分子被成功地合成了在胎圈，但裂解过程中降解。当如上述那样低的温度条件下裂解，三氟乙酸诱导的降解的量被大大地抑制， 如图6B所示 。这样的裂解的机制已...

Watch this Scientific Journal Video about Split-and-pool Synthesis and Characterization of Peptide Tertiary Amide Library at JoVE.com

Split-and-pool Synthesis and Characterization of Peptide Tertiary Amide Library

肽叔酰胺（贸易协定）的肽模拟物，包括但不限于肽，拟肽和N-甲基化的肽的一个家族。在这里，我们描述它结合了两个分裂和池和子策略单体合成家教会的一珠一化合物库的合成方法。

分裂和池合成及肽叔酰胺图书馆表征

Peptidomimetics are great sources of protein ligands. The oligomeric nature of these compounds enables us to access large synthetic libraries on solid ...

split-pool-synthesis-characterization-peptide-tertiary-amide

Research

Education

JoVE Core

JoVE Journal

Organic Chemistry

Chemistry

Unit 1

Carboxylic Acid Derivatives

SCIENTISTS IN ACTION

18.0K 次观看.  The Scripps Research Institute. 肽叔酰胺（贸易协定）的肽模拟物，包括但不限于肽，拟肽和N-甲基化的肽的一个家族。在这里，我们描述它结合了两个分裂和池和子策略单体合成家教会的一珠一化合物库的合成方法。 

视频: Split-and-pool Synthesis and Characterization of Peptide Tertiary Amide Library

Nucleoside Triphosphates - From Synthesis to Biochemical Characterization

The traditional strategy for the introduction of chemical functionalities is the use of solid-phase synthesis by appending suitably modified phosphoramidite precursors to the nascent chain. However, the conditions used during the synthesis and the restriction to rather short sequences hamper the applicability of this methodology. On the other hand, modified nucleoside triphosphates are activated building blocks that have been employed for the mild introduction of numerous functional groups into nucleic acids, a strategy that paves the way for the use of modified nucleic acids in a wide-ranging palette of practical applications such as functional tagging and generation of ribozymes and DNAzymes. One of the major challenges resides in the intricacy of the methodology leading to the isolation and characterization of these nucleoside analogues.
	
	In this video article, we present a detailed protocol for the synthesis of these modified analogues using phosphorous(III)-based reagents. In addition, the procedure for their biochemical characterization is divulged, with a special emphasis on primer extension reactions and TdT tailing polymerization. This detailed protocol will be of use for the crafting of modified dNTPs and their further use in chemical biology.

The protocol described herein aims to explain and abridge the numerous obstacles in the way of the intricate route leading to modified nucleoside triphosphates. Consequently, this protocol facilitates both the synthesis of these activated building-blocks and their availability for practical applications.

核苷三磷酸 - 从合成到生化特征

The  traditional strategy for the introduction of chemical functionalities is the  use of solid-phase synthesis by appending suitably modified ...

科研

化学

Amide Coupling Reaction for the Synthesis of Bispyridine-based Ligands and Their Complexation to Platinum as Dinuclear Anticancer Agents

Amide coupling reactions can be used to synthesize bispyridine-based ligands for use as bridging linkers in multinuclear platinum anticancer drugs. Isonicotinic acid, or its derivatives, are coupled to variable length diaminoalkane chains under an inert atmosphere in anhydrous DMF or DMSO with the use of a weak base, triethylamine, and a coupling agent, 1-propylphosphonic anhydride. The products precipitate from solution upon formation or can be precipitated by the addition of water. If desired, the ligands can be further purified by recrystallization from hot water. Dinuclear platinum complex synthesis using the bispyridine ligands is done in hot water using transplatin. The most informative of the chemical characterization techniques to determine the structure and gross purity of both the bispyridine ligands and the final platinum complexes is 1H NMR with particular analysis of the aromatic region of the spectra (7-9 ppm). The platinum complexes have potential application as anticancer agents and the synthesis method can be modified to produce trinuclear and other multinuclear complexes with different hydrogen bonding functionality in the bridging ligand.

This protocol describes the use of amide coupling reactions of isonicotinic acid and diaminoalkanes to form bridging ligands suitable for use in the synthesis of multinuclear platinum complexes, which combine aspects of the anticancer drugs BBR3464 and picoplatin.

酰胺偶联反应为双吡啶基配体及其络合合成，以白金为双核抗癌药物

Amide coupling reactions can be used to synthesize bispyridine-based ligands for use as bridging linkers in multinuclear platinum anticancer drugs. ...

Synthesis and Characterization of Functionalized Metal-organic Frameworks

Metal-organic frameworks have attracted extraordinary amounts of research attention, as they are attractive candidates for numerous industrial and technological applications. Their signature property is their ultrahigh porosity, which however imparts a series of challenges when it comes to both constructing them and working with them. Securing desired MOF chemical and physical functionality by linker/node assembly into a highly porous framework of choice can pose difficulties, as less porous and more thermodynamically stable congeners (e.g., other crystalline polymorphs, catenated analogues) are often preferentially obtained by conventional synthesis methods. Once the desired product is obtained, its characterization often requires specialized techniques that address complications potentially arising from, for example, guest-molecule loss or preferential orientation of microcrystallites. Finally, accessing the large voids inside the MOFs for use in applications that involve gases can be problematic, as frameworks may be subject to collapse during removal of solvent molecules (remnants of solvothermal synthesis). In this paper, we describe synthesis and characterization methods routinely utilized in our lab either to solve or circumvent these issues. The methods include solvent-assisted linker exchange, powder X-ray diffraction in capillaries, and materials activation (cavity evacuation) by supercritical CO2 drying. Finally, we provide a protocol for determining a suitable pressure region for applying the Brunauer-Emmett-Teller analysis to nitrogen isotherms, so as to estimate surface area of MOFs with good accuracy.

Synthesis, activation, and characterization of intentionally designed metal-organic framework materials is challenging, especially when building blocks are incompatible or unwanted polymorphs are thermodynamically favored over desired forms. We describe how applications of solvent-assisted linker exchange, powder X-ray diffraction in capillaries and activation via supercritical CO2 drying, can address some of these challenges.

功能化的合成与表征金属 - 有机骨架

Metal-organic frameworks have attracted extraordinary amounts of research attention, as they are attractive candidates for numerous industrial and ...

The Synthesis, Characterization and Reactivity of a Series of Ruthenium N-triphosPh Complexes

Herein we report the synthesis of a tridentate phosphine ligand N(CH2PPh2)3 (N-triphosPh) (1) via a phosphorus based Mannich reaction of the hydroxylmethylene phosphine precursor with ammonia in methanol under a nitrogen atmosphere. The N-triphosPh ligand precipitates from the solution after approximately 1 hr of reflux and can be isolated analytically pure via simple cannula filtration procedure under nitrogen. Reaction of the N-triphosPh ligand with [Ru3(CO)12] under reflux affords a deep red solution that show evolution of CO gas on ligand complexation. Orange crystals of the complex [Ru(CO)2{N(CH2PPh2)3}-&#954;3P] (2) were isolated on cooling to RT. The 31P{1H} NMR spectrum showed a characteristic single peak at lower frequency compared to the free ligand. Reaction of a toluene solution of complex 2 with oxygen resulted in the instantaneous precipitation of the carbonate complex [Ru(CO3)(CO){N(CH2PPh2)3}-&#954;3P] (3) as an air stable orange solid. Subsequent hydrogenation of 3 under 15 bar of hydrogen in a high-pressure reactor gave the dihydride complex [RuH2(CO){N(CH2PPh2)3}-&#954;3P] (4), which was fully characterized by X-ray crystallography and NMR spectroscopy. Complexes 3 and 4 are potentially useful catalyst precursors for a range of hydrogenation reactions, including biomass-derived products such as levulinic acid (LA). Complex 4 was found to cleanly react with LA in the presence of the proton source additive NH4PF6 to give [Ru(CO){N(CH2PPh2)3}-&#954;3P{CH3CO(CH2)2CO2H}-&#954;2O](PF6) (6).

The Synthesis, Characterization and Reactivity of a Series of Ruthenium N-triphosPh Complexes

Ruthenium phosphine complexes are widely used for homogeneous catalytic reactions such as hydrogenations. The synthesis of a series of novel tridentate ruthenium complexes bearing the N-triphos ligand N(CH2PPh2)3 is reported. Additionally, the stoichiometric reaction of a dihydride Ru&#8211;N-triphos complex with levulinic acid is described.

一系列钌的合成，表征和反应<em&gt;Ñ</em&gt; -triphos<sup&gt;博士</sup&gt;配合

Herein we report the synthesis of a tridentate phosphine ligand N(CH2PPh2)3 (N-triphosPh) (1) via a phosphorus based Mannich reaction of the ...

Synthesis, Characterization, and Functionalization of Hybrid Au/CdS and Au/ZnS Core/Shell Nanoparticles

Plasmonic nanoparticles are an attractive material for light harvesting applications due to their easily modified surface, high surface area and large extinction coefficients which can be tuned across the visible spectrum. Research into the plasmonic enhancement of optical transitions has become popular, due to the possibility of altering and in some cases improving photo-absorption or emission properties of nearby chromophores such as molecular dyes or quantum dots. The electric field of the plasmon can couple with the excitation dipole of a chromophore, perturbing the electronic states involved in the transition and leading to increased absorption and emission rates. These enhancements can also be negated at close distances by energy transfer mechanism, making the spatial arrangement of the two species critical. Ultimately, enhancement of light harvesting efficiency in plasmonic solar cells could lead to thinner and, therefore, lower cost devices. The development of hybrid core/shell particles could offer a solution to this issue. The addition of a dielectric spacer between a gold nanoparticles and a chromophore is the proposed method to control the exciton plasmon coupling strength and thereby balance losses with the plasmonic gains. A detailed procedure for the coating of gold nanoparticles with CdS and ZnS semiconductor shells is presented. The nanoparticles show high uniformity with size control in both the core gold particles and shell species allowing for a more accurate investigation into the plasmonic enhancement of external chromophores.

The synthesis of uniform gold nanoparticles coated with semiconductor shells of CdS or ZnS is performed. The semiconductor coating is conducted by first depositing a silver sulfide shell and exchanging the silver cations for zinc or cadmium cations.

合成，表征和混合金/硫化镉和Au / ZnS核/壳纳米颗粒功能化

Plasmonic nanoparticles are an attractive material for light harvesting applications due to their easily modified surface, high surface area and large ...

Development of a Backbone Cyclic Peptide Library as Potential Antiparasitic Therapeutics Using Microwave Irradiation

Protein-protein interactions (PPIs) are intimately involved in almost all biological processes and are linked to many human diseases. Therefore, there is a major effort to target PPIs in basic research and in the pharmaceutical industry. Protein-protein interfaces are usually large, flat, and often lack pockets, complicating the discovery of small molecules that target such sites. Alternative targeting approaches using antibodies have limitations due to poor oral bioavailability, low cell-permeability, and production inefficiency.
Using peptides to target PPI interfaces has several advantages. Peptides have higher conformational flexibility, increased selectivity, and are generally inexpensive. However, peptides have their own limitations including poor stability and inefficiency crossing cell membranes. To overcome such limitations, peptide cyclization can be performed. Cyclization has been demonstrated to improve peptide selectivity, metabolic stability, and bioavailability. However, predicting the bioactive conformation of a cyclic peptide is not trivial. To overcome this challenge, one attractive approach it to screen a focused library to screen in which all backbone cyclic peptides have the same primary sequence, but differ in parameters that influence their conformation, such as ring size and position.
We describe a detailed protocol for synthesizing a library of backbone cyclic peptides targeting specific parasite PPIs. Using a rational design approach, we developed peptides derived from the scaffold protein Leishmania receptor for activated C-kinase (LACK). We hypothesized that sequences in LACK that are conserved in parasites, but not in the mammalian host homolog, may represent interaction sites for proteins that are critical for the parasites&#39; viability. The cyclic peptides were synthesized using microwave irradiation to reduce reaction times and increase efficiency. Developing a library of backbone cyclic peptides with different ring sizes facilitates a systematic screen for the most biological active conformation. This method provides a general, fast, and facile way to synthesize cyclic peptides.

A simple and general method for the synthesis of cyclic peptides using microwave irradiation is outlined. This procedure enables the synthesis of backbone cyclic peptides with a collection of different conformations while retaining the side chains and the pharmacophoric moieties., and therefore, allows to screen for the bioactive conformation.

一个主干环肽库作为潜在的抗寄生虫治疗微波辐射的发展

Protein-protein interactions (PPIs) are intimately involved in almost all biological processes and are linked to many human diseases. Therefore, there is ...

Synthesis and Characterization of Supramolecular Colloids

Control over colloidal assembly is of utmost importance for the development of functional colloidal materials with tailored structural and mechanical properties for applications in photonics, drug delivery and coating technology. Here we present a new family of colloidal building blocks, coined supramolecular colloids, whose self-assembly is controlled through surface-functionalization with a benzene-1,3,5-tricarboxamide (BTA) derived supramolecular moiety. Such BTAs interact via directional, strong, yet reversible hydrogen-bonds with other identical BTAs. Herein, a protocol is presented that describes how to couple these BTAs to colloids and how to quantify the number of coupling sites, which determines the multivalency of the supramolecular colloids. Light scattering measurements show that the refractive index of the colloids is almost matched with that of the solvent, which strongly reduces the van der Waals forces between the colloids. Before photo-activation, the colloids remain well dispersed, as the BTAs are equipped with a photo-labile group that blocks the formation of hydrogen-bonds. Controlled deprotection with UV-light activates the short-range hydrogen-bonds between the BTAs, which triggers the colloidal self-assembly. The evolution from the dispersed state to the clustered state is monitored by confocal microscopy. These results are further quantified by image analysis with simple routines using ImageJ and Matlab. This merger of supramolecular chemistry and colloidal science offers a direct route towards light- and thermo-responsive colloidal assembly encoded in the surface-grafted monolayer.

A protocol for the synthesis and characterization of colloids coated with supramolecular moieties is described. These supramolecular colloids undergo self-assembly upon the activation of the hydrogen-bonds between the surface-anchored molecules by UV-light.

超分子胶体的合成与表征

Control over colloidal assembly is of utmost importance for the development of functional colloidal materials with tailored structural and mechanical ...

Synthesis and Characterization of Fe-doped Aluminosilicate Nanotubes with Enhanced Electron Conductive Properties

The goal of the protocol is to synthesize Fe-doped aluminosilicate nanotubes of the imogolite type with the formula (OH)3Al2-xFexO3SiOH. Doping with Fe aims at lowering the band gap of imogolite, an insulator with the chemical formula (OH)3Al2O3SiOH, and at modifying its adsorption properties towards azo-dyes, an important class of organic pollutants of both wastewater and groundwater.
Fe-doped nanotubes are obtained in two ways: by direct synthesis, where FeCl3 is added to an aqueous mixture of the Si and Al precursors, and by post-synthesis loading, where preformed nanotubes are put in contact with a FeCl3&#8226;6H2O aqueous solution. In both synthesis methods, isomorphic substitution of Al3+ by Fe3+ occurs, preserving the nanotube structure. Isomorphic substitution is indeed limited to a mass fraction of ~1.0% Fe, since at a higher Fe content (i.e., a mass fraction of 1.4% Fe), Fe2O3 clusters form, especially when the loading procedure is adopted. The physicochemical properties of the materials are studied by means of X-ray powder diffraction (XRD), N2 sorption isotherms at -196 &#176;C, high resolution transmission electron microscopy (HRTEM), diffuse reflectance (DR) UV-Vis spectroscopy, and &#950;-potential measurements. The most relevant result is the possibility to replace Al3+ ions (located on the outer surface of the nanotubes) by post-synthesis loading on preformed imogolite without perturbing the delicate hydrolysis equilibria occurring during nanotube formation. During the loading procedure, an anionic exchange occurs, where Al3+ ions on the outer surface of the nanotubes are replaced by Fe3+ ions. In Fe-doped aluminosilicate nanotubes, isomorphic substitution of Al3+ by Fe3+ is found to affect the band gap of doped imogolite. Nonetheless, Fe3+ sites on the outer surface of nanotubes are able to coordinate organic moieties, like the azo-dye Acid Orange 7, through a ligand-displacement mechanism occurring in an aqueous solution.

Here, we present a protocol to synthesize and characterize Fe-doped aluminosilicate nanotubes. The materials are obtained by either sol-gel synthesis upon addition of FeCl3&#8226;6H2O to the mixture containing the Si and Al precursors or by post-synthesis ionic exchange of preformed aluminosilicate nanotubes.

以增强电子导电性能的合成与表征Fe掺杂碳纳米管硅铝的

The goal of the protocol is to synthesize Fe-doped aluminosilicate nanotubes of the imogolite type with the formula (OH)3Al2-xFexO3SiOH. Doping with Fe ...

Facile Protocol for the Synthesis of Self-assembling Polyamine-based Peptide Amphiphiles (PPAs) and Related Biomaterials

Polyamine-based Peptide Amphiphiles (PPAs) are a new class of self-assembling amphiphilic biomaterials-related to the peptide amphiphiles (PAs). Traditional PAs possess charged amino acids as solubilizing groups (lysine, arginine), which are directly connected to a lipid segment or can contain a linker region made of neutral amino acids. Tuning the peptide sequence of PAs can yield diverse morphologies. Similarly, PPAs possess a hydrophobic segment and neutral amino acids, but also contain polyamine molecules as water solubilizing (hydrophilic) groups. As is the case with PAs, PPAs can also self-assemble into diverse morphologies, including small rods, twisted nano-ribbons, and fused nano-sheets, when dissolved in water. However, the presence of both primary and secondary amines on a single polyamine molecule poses a significant challenge when synthesizing PPAs. In this paper, we show a simple protocol, based on literature precedents, to achieve a facile synthesis of PPAs using solid phase peptide synthesis (SPPS). This protocol can be extended to the synthesis of PAs and other similar systems. We also illustrate the steps that are needed for cleavage from the resin, identification, and purification.

Facile Protocol for the Synthesis of Self-assembling Polyamine-based Peptide Amphiphiles PPAs and Related Biomaterials

The synthesis of polyamine-based peptide amphiphiles (PPAs) is a significant challenge due to the presence of multiple amine nitrogens, which requires judicious use of protecting groups to mask these reactive functionalities. In this paper, we describe a facile method for the preparation of these new class of self-assembling molecules.

自组装多胺基多肽 Amphiphiles (PPAs) 及相关生物材料的简易合成协议

Polyamine-based Peptide Amphiphiles (PPAs) are a new class of self-assembling amphiphilic biomaterials-related to the peptide amphiphiles (PAs). ...

Synthesis and Characterization of 1,2-Dithiolane Modified Self-Assembling Peptides

This report focuses on the synthesis of an N-terminus 1,2-dithiolane modified self-assembling peptide and the characterization of the resulting self-assembled supramolecular structures. The synthetic route takes advantage of solid-phase peptide synthesis with the on-resin coupling of the dithiolane precursor molecule, 3-(acetylthio)-2-(acetylthiomethyl)propanoic acid, and the microwave-assisted thioacetate deprotection of the peptide N-terminus before final cleavage from the resin to yield the 1,2-dithiolane modified peptide. After the high-performance liquid chromatography (HPLC) purification of the 1,2-dithiolane peptide, derived from the nucleating core of the A&#946; peptide associated with Alzheimer&#39;s disease, the peptide is shown to self-assemble into cross-&#946; amyloid fibers. Protocols to characterize the amyloid fibers by Fourier-transform infrared spectroscopy (FT-IR), circular dichroism spectroscopy (CD) and transmission electron microscopy (TEM) are presented. The methods of N-terminal modification with a 1,2-dithiolane moiety to well-characterized self-assembling peptides can now be explored as model systems to develop post-assembly modification strategies and explore dynamic covalent chemistry on supramolecular peptide nanofiber surfaces.

A protocol for the synthesis of a 1,2-dithiolane modified peptide and the characterization of the supramolecular structures resulting from the peptide self-assembly.