400-998-5282
专注多肽 服务科研
400-998-5282
专注多肽 服务科研
可显着逆转小鼠低水平的吗啡镇痛感受性。
编号：156270
CAS号：121932-06-7
单字母：H2N-TYADFIASGRTGRRNAI-CONH2
| 参考文献（References）: | D.B. Glass et al., JBC, 264, 1457 (1989) 1. Glass, D.B., Cheng, H.C., Mende-Mueller, L., et al . Primary structural determinants essential for potent inhibition of cAMP- dependent protein kinase by inhibitory peptides corresponding to the active portion of the heat- stable inhibitor protein . The Journal of Biological Chemisty 264(15), 8802-8810 (1989) . <ALLPEPTIDE>2. Glass, D.B., Lundquist, L.J., Katz, B.M., et al . Protein kinase inhibitor- (6- 22)- amide peptide analogs with standard and nonstandard amino acid substitutions for phenylalanine10. Inhibition of cAMP- dependent protein kinase . The Journal of Biological Chemisty 264(24), 14579-14584 (1989) . 1. Glass, D.B., Cheng, H.C., Mende-Mueller, L., et al . Primary structural determinants essential for potent inhibition of cAMP- dependent protein kinase by inhibitory peptides corresponding to the active portion of the heat- stable inhibitor protein . The Journal of Biological Chemisty 264(15), 8802-8810 (1989) . <ALLPEPTIDE>2. Glass, D.B., Lundquist, L.J., Katz, B.M., et al . Protein kinase inhibitor- (6- 22)- amide peptide analogs with standard and nonstandard amino acid substitutions for phenylalanine10. Inhibition of cAMP- dependent protein kinase . The Journal of Biological Chemisty 264(24), 14579-14584 (1989) . |
| 溶解度: | DMF: 20 mg/ml DMSO: 30 mg/ml PBS (pH 7.2): 10 mg/ml |
| SMILES: | NC([C@@]([C@H](CC)C)([H])NC([C@@H](NC([C@H](CC(N)=O)NC([C@H](CCCNC(N)=N)NC([C@H](CCCNC(N)=N)NC(CNC([C@@]([C@@H](C)O)([H])NC([C@H](CCCNC(N)=N)NC(CNC([C@H](CO)NC([C@@H](NC([C@@]([C@H](CC)C)([H])NC([C@@H](NC([C@@H](NC([C@@H](NC([C@@H](NC([C@@]([C@@H](C)O)([H])N[H])=O)CC1=CC=C(O)C=C1)=O)C)=O)CC(O)=O)=O)CC2=CC=CC=C2)=O)=O)C)=O)=O)=O)=O)=O)=O)=O)=O)=O)C)=O)=O |
| InChI/InChI Code: | InChI=1S/C80H130N28O24/c1-10-37(3)60(63(83)118)106-66(121)41(7)96-72(127)52(32-55(81)113)103-69(124)49(22-17-29-92-80(88)89)100-68(123)47(20-15-27-90-78(84)85)98-57(115)35-94-76(131)62(43(9)111)108-70(125)48(21-16-28-91-79(86)87)99-56(114)34-93-67(122)54(36-109)105-65(120)40(6)97-77(132)61(38(4)11-2)107-74(129)51(30-44-18-13-12-14-19-44)102-73(128)53(33-58(116)117)101-64(119)39(5)95-71(126)50(104-75(130)59(82)42(8)110)31-45-23-25-46(112)26-24-45/h12-14,18-19,23-26,37-43,47-54,59-62,109-112H,10-11,15-17,20-22,27-36,82H2,1-9H3,(H2,81,113)(H2,83,118)(H,93,122)(H,94,131)(H,95,126)(H,96,127)(H,97,132)(H,98,115)(H,99,114)(H,100,123)(H,101,119)(H,102,128)(H,103,124)(H,104,130)(H,105,120)(H,106,121)(H,107,129)(H,108,125)(H,116,117)(H4,84,85,90)(H4,86,87,91)(H4,88,89,92)/t37-,38-,39-,40-,41-,42+,43+,47-,48-,49-,50-,51-,52-,53-,54-,59-,60-,61-,62-/m0/s1 |
| InChI Key: | VAKHFAFLRUNHLQ-PEBJKXEYSA-N |
| Formal Name: | L-threonyl-L-tyrosyl-L-alanyl-L-α-aspartyl-L-phenylalanyl-L-isoleucyl-L-alanyl-L-serylglycyl-L-arginyl-L-threonylglycyl-L-arginyl-L-arginyl-L-asparaginyl-L-alanyl-L-isoleucinamide |
PKA Inhibitor Fragment (6-22) amide 是 cAMP 依赖性蛋白激酶 A (PKA) 的抑制剂，Ki 值为 2.8 nM。PKA Inhibitor Fragment (6-22) amide 可显着逆转小鼠低水平的吗啡镇痛感受性。
PKA Inhibitor Fragment (6-22) amide is an inhibitor of cAMP-dependent protein kinase A (PKA), with a Ki of 2.8 nM. PKA Inhibitor Fragment (6-22) amide can significantly reverse low-level morphine antinociceptive tolerance in mice.
PKA抑制剂片段（6-22）酰胺乙酸酯是cAMP依赖性蛋白激酶（PKA）（Ki=2.5nM）的合成肽抑制剂，来源于热稳定的PKA抑制剂蛋白PKI。它是最短的合成PKI肽，对PKA的抑制具有很高的效力。
PKA Inhibitor Fragment (6-22) amide acetate is a synthetic peptide inhibitor of cAMP-dependent protein kinase (PKA) (Ki = 2.5 nM), derived from the heat-stable PKA inhibitor protein PKI. It is the shortest synthetic PKI peptide that retains high potency for PKA inhibition.
环磷酸腺苷依赖性蛋白激酶抑制肽 6 至 22 位酰胺片段，是由热稳定抑制蛋白改造而来的合成抑制多肽，也是保留强效抑制活性的最短肽段，依靠两端特定氨基酸结构实现高亲和力结合，发挥酶抑制作用。
PKA inhibitor fragment (6-22) amide is a synthetic peptide inhibitor of cAMP-dependent protein kinase (PKA; K = 1.7 nM) derived from the heat-stable PKA inhibitor protein PKI. It is the shortest synthetic PKI peptide that retains high potency for PKA inhibition. Both the arginine-containing pseudosubstrate site of the PKI peptide in its COOH terminus and the residue Phe in NH -terminal portion are required for this high affinity binding.
环磷酸腺苷依赖性蛋白激酶抑制肽 6 至 22 位酰胺片段，是由热稳定抑制蛋白改造而来的合成抑制多肽，也是保留强效抑制活性的最短肽段，依靠两端特定氨基酸结构实现高亲和力结合，发挥酶抑制作用。
PKA inhibitor fragment (6-22) amide is a synthetic peptide inhibitor of cAMP-dependent protein kinase (PKA; K = 1.7 nM) derived from the heat-stable PKA inhibitor protein PKI. It is the shortest synthetic PKI peptide that retains high potency for PKA inhibition. Both the arginine-containing pseudosubstrate site of the PKI peptide in its COOH terminus and the residue Phe in NH -terminal portion are required for this high affinity binding.
Katz BM, et, al. Synthesis, characterization and inhibitory activities of (4-N3[3,5-3H]Phe10)PKI(6-22)amide and its precursors: photoaffinity labeling peptides for the active site of cyclic AMP-dependent protein kinase. Int J Pept Protein Res. 1989 Jun;33(6):439 : https://pubmed.ncbi.nlm.nih.gov/2550380/
Dalton GD, et, al. Alterations in brain Protein Kinase A activity and reversal of morphine tolerance by two fragments of native Protein Kinase A inhibitor peptide (PKI). Neuropharmacology. 2005 Apr; 48(5): 648-57. : https://pubmed.ncbi.nlm.nih.gov/15814100/
多肽H2N-Thr-Tyr-Ala-Asp-Phe-Ile-Ala-Ser-Gly-Arg-Thr-Gly-Arg-Arg-Asn-Ala-Ile-NH2的合成步骤：
1、合成MBHA树脂：取若干克的MBHA树脂（如初始取代度为0.5mmol/g）和1倍树脂摩尔量的Fmoc-Linker-OH加入到反应器中，加入DMF，搅拌使氨基酸完全溶解。再加入树脂2倍量的DIEPA，搅拌混合均匀。再加入树脂0.95倍量的HBTU，搅拌混合均匀。反应3-4小时后，用DMF洗涤3次。用2倍树脂体积的10%乙酸酐/DMF 进行封端30分钟。然后再用DMF洗涤3次，甲醇洗涤2次，DCM洗涤2次，再用甲醇洗涤2次。真空干燥12小时以上，得到干燥的树脂{Fmoc-Linker-MHBA Resin}，测定取代度。这里测得取代度为 0.3mmol/g。结构如下图：
2、脱Fmoc：取2.67g的上述树脂，用DCM或DMF溶胀20分钟。用DMF洗涤2遍。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-Linker-MBHA 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2.4mmol Fmoc-Ile-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4.81mmol DIPEA，2.28mmol 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-Ile-Linker-MBHA 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-Ile-Linker-MBHA Resin
Fmoc-Ala-Ile-Linker-MBHA Resin
H2N-Ala-Ile-Linker-MBHA Resin
Fmoc-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Ile-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Ile-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Phe-Ile-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Phe-Ile-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Asp(OtBu)-Phe-Ile-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Asp(OtBu)-Phe-Ile-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Ala-Asp(OtBu)-Phe-Ile-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Ala-Asp(OtBu)-Phe-Ile-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Tyr(tBu)-Ala-Asp(OtBu)-Phe-Ile-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
H2N-Tyr(tBu)-Ala-Asp(OtBu)-Phe-Ile-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA Resin
Fmoc-Thr(tBu)-Tyr(tBu)-Ala-Asp(OtBu)-Phe-Ile-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA 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-Thr(tBu)-Tyr(tBu)-Ala-Asp(OtBu)-Phe-Ile-Ala-Ser(tBu)-Gly-Arg(Pbf)-Thr(tBu)-Gly-Arg(Pbf)-Arg(Pbf)-Asn(Trt)-Ala-Ile-Linker-MBHA 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-Thr-Tyr-Ala-Asp-Phe-Ile-Ala-Ser-Gly-Arg-Thr-Gly-Arg-Arg-Asn-Ala-Ile-NH2。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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