400-998-5282
专注多肽 服务科研
400-998-5282
专注多肽 服务科研
构建了特异性结合核仁素的多肽探针FITC-KDEPQRRSARLSAKPAPPK- PEPKPKKAPAKK
编号：411573
CAS号：
单字母：H2N-KDEPQRRSARLSAKPAPPKPEPKPKKAPAKK-OH
在肿瘤微环境中, 肿瘤血管的生成是肿瘤生长和转移的重要条件. 肿瘤血管内皮细胞通过分泌血管生成因子, 促进肿瘤血管的生成及肿瘤细胞的粘附和转移. 核仁素作为肿瘤内皮细胞的标志物, 是检测肿瘤血管新生的重要靶点. Ruoslahti等构建了特异性结合核仁素的多肽探针FITC-KDEPQRRSARLSAKPAPPK- PEPKPKKAPAKK, 同时, Ying等利用相同的靶向多肽构建了CdTe量子点-多肽探针, 用于人乳腺癌细胞MDA-MB-435的活细胞成像.
靶向多肽可以根据其功能和用途分为不同的类别。在PDC（多肽偶联药物）中，靶向多肽通常被分为细胞穿透肽和细胞靶向肽两大类。
细胞穿透肽：这类多肽能够跨越细胞膜，转运具有生物活性的大分子物质，如多肽、蛋白质、核酸等化学药物，使其顺利进入细胞。一些常见的细胞穿透肽包括Pep-1、Pentratin、PepFact14、Transportan等。
细胞靶向肽：这类多肽的作用主要是引导化学药物或生物活性分子与特定类型的细胞结合，以提高其靶向性和治疗效率。常见的细胞靶向肽包括PEGA、生长激素抑制素类似物、蛙皮素类似物、RGD肽类等。
多肽H2N-Lys-Asp-Glu-Pro-Gln-Arg-Arg-Ser-Ala-Arg-Leu-Ser-Ala-Lys-Pro-Ala-Pro-Pro-Lys-Pro-Glu-Pro-Lys-Pro-Lys-Lys-Ala-Pro-Ala-Lys-Lys-COOH的合成步骤：
1、合成CTC树脂：称取1.09g CTC Resin（如初始取代度约为0.51mmol/g）和0.67mmol Fmoc-Lys(Boc)-OH于反应器中，加入适量DCM溶解氨基酸（需要注意，此时CTC树脂体积会增大好几倍，避免DCM溶液过少），再加入1.67mmol DIPEA（Mw：129.1,d：0.740g/ml），反应2-3小时后，可不抽滤溶液，直接加入1ml的HPLC级甲醇，封端半小时。依次用DMF洗涤2次，甲醇洗涤1次，DCM洗涤一次，甲醇洗涤一次，DCM洗涤一次，DMF洗涤2次（这里使用甲醇和DCM交替洗涤，是为了更好地去除其他溶质，有利于后续反应）。得到 Fmoc-Lys(Boc)-CTC Resin。结构图如下：
2、脱Fmoc：加3倍树脂体积的20%Pip/DMF溶液，鼓氮气30分钟，然后2倍树脂体积的DMF 洗涤5次。得到 H2N-Lys(Boc)-CTC Resin 。（此步骤脱除Fmoc基团，茚三酮检测为蓝色，Pip为哌啶）。结构图如下：
3、缩合：取1.67mmol Fmoc-Lys(Boc)-OH 氨基酸，加入到上述树脂里，加适当DMF溶解氨基酸，再依次加入3.34mmol DIPEA，1.58mmol HBTU。反应30分钟后，取小样洗涤，茚三酮检测为无色。用2倍树脂体积的DMF 洗涤3次树脂。（洗涤树脂，去掉残留溶剂，为下一步反应做准备）。得到Fmoc-Lys(Boc)-Lys(Boc)-CTC Resin。氨基酸：DIPEA：HBTU：树脂=3：6：2.85：1（摩尔比）。结构图如下：
4、依次循环步骤二、步骤三，依次得到
H2N-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Arg(Pbf)-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Arg(Pbf)-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Pro-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Pro-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Glu(OtBu)-Pro-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Glu(OtBu)-Pro-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Asp(OtBu)-Glu(OtBu)-Pro-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
H2N-Asp(OtBu)-Glu(OtBu)-Pro-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
Fmoc-Lys(Boc)-Asp(OtBu)-Glu(OtBu)-Pro-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin
以上中间结构，均可在专肽生物多肽计算器-多肽结构计算器中，一键画出。
最后再经过步骤二得到 H2N-Lys(Boc)-Asp(OtBu)-Glu(OtBu)-Pro-Gln(Trt)-Arg(Pbf)-Arg(Pbf)-Ser(tBu)-Ala-Arg(Pbf)-Leu-Ser(tBu)-Ala-Lys(Boc)-Pro-Ala-Pro-Pro-Lys(Boc)-Pro-Glu(OtBu)-Pro-Lys(Boc)-Pro-Lys(Boc)-Lys(Boc)-Ala-Pro-Ala-Lys(Boc)-Lys(Boc)-CTC Resin，结构如下：
5、切割：6倍树脂体积的切割液（或每1g树脂加8ml左右的切割液），摇床摇晃 2小时，过滤掉树脂，用冰无水乙醚沉淀滤液，并用冰无水乙醚洗涤沉淀物3次，最后将沉淀物放真空干燥釜中，常温干燥24小试，得到粗品H2N-Lys-Asp-Glu-Pro-Gln-Arg-Arg-Ser-Ala-Arg-Leu-Ser-Ala-Lys-Pro-Ala-Pro-Pro-Lys-Pro-Glu-Pro-Lys-Pro-Lys-Lys-Ala-Pro-Ala-Lys-Lys-COOH。结构图见产品结构图。
切割液选择：1）TFA：H2O=95%：5%
2）TFA：H2O：TIS=95%：2.5%：2.5%
3）三氟乙酸：茴香硫醚：1，2-乙二硫醇：苯酚：水=87.5%：5%：2.5%：2.5%：2.5%
（前两种适合没有容易氧化的氨基酸，例如Trp、Cys、Met。第三种适合几乎所有的序列。）
6、纯化冻干：使用液相色谱纯化，收集目标峰液体，进行冻干，获得蓬松的粉末状固体多肽。不过这时要取小样复测下纯度 是否目标纯度。
7、最后总结：
杭州专肽生物技术有限公司（ALLPEPTIDE https://www.allpeptide.com）主营定制多肽合成业务，提供各类长肽，短肽，环肽，提供各类修饰肽，如：荧光标记修饰（CY3、CY5、CY5.5、CY7、FAM、FITC、Rhodamine B、TAMRA等），功能基团修饰肽（叠氮、炔基、DBCO、DOTA、NOTA等），同位素标记肽（N15、C13），订书肽（Stapled Peptide）,脂肪酸修饰肽（Pal、Myr、Ste），磷酸化修饰肽（P-Ser、P-Thr、P-Tyr），环肽（酰胺键环肽、一对或者多对二硫键环），生物素标记肽，PEG修饰肽，甲基化修饰肽等。
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