CN103864885B - The application of 1-hydroxyl-1,2,3-phentriazine-4 (3H)-one in Peptide systhesis - Google Patents
The application of 1-hydroxyl-1,2,3-phentriazine-4 (3H)-one in Peptide systhesis Download PDFInfo
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- CN103864885B CN103864885B CN201410107410.6A CN201410107410A CN103864885B CN 103864885 B CN103864885 B CN 103864885B CN 201410107410 A CN201410107410 A CN 201410107410A CN 103864885 B CN103864885 B CN 103864885B
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 59
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 50
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 32
- 229920001184 polypeptide Polymers 0.000 claims abstract description 29
- 150000001413 amino acids Chemical class 0.000 claims abstract description 26
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims abstract description 24
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 16
- 239000003960 organic solvent Substances 0.000 claims abstract description 15
- -1 carbobenzoxy-(Cbz) Chemical class 0.000 claims abstract description 14
- 150000004702 methyl esters Chemical class 0.000 claims abstract description 9
- 150000007524 organic acids Chemical class 0.000 claims abstract description 5
- 150000003839 salts Chemical class 0.000 claims abstract description 5
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 120
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 58
- 238000006243 chemical reaction Methods 0.000 claims description 56
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 49
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 48
- 239000000243 solution Substances 0.000 claims description 39
- 239000011734 sodium Substances 0.000 claims description 30
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 27
- 238000003756 stirring Methods 0.000 claims description 27
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical group C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 26
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 18
- 238000004821 distillation Methods 0.000 claims description 18
- 239000005457 ice water Substances 0.000 claims description 18
- 239000012074 organic phase Substances 0.000 claims description 18
- 238000002360 preparation method Methods 0.000 claims description 18
- FYSNRJHAOHDILO-UHFFFAOYSA-N thionyl chloride Chemical compound ClS(Cl)=O FYSNRJHAOHDILO-UHFFFAOYSA-N 0.000 claims description 18
- 239000013078 crystal Substances 0.000 claims description 16
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 13
- 235000002639 sodium chloride Nutrition 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 238000006482 condensation reaction Methods 0.000 claims description 10
- 238000000926 separation method Methods 0.000 claims description 10
- 239000007787 solid Substances 0.000 claims description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical class [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 9
- HSDAJNMJOMSNEV-UHFFFAOYSA-N benzyl chloroformate Chemical compound ClC(=O)OCC1=CC=CC=C1 HSDAJNMJOMSNEV-UHFFFAOYSA-N 0.000 claims description 9
- 238000004440 column chromatography Methods 0.000 claims description 9
- 229960004132 diethyl ether Drugs 0.000 claims description 9
- IXCSERBJSXMMFS-UHFFFAOYSA-N hydrogen chloride Substances Cl.Cl IXCSERBJSXMMFS-UHFFFAOYSA-N 0.000 claims description 9
- 229910000041 hydrogen chloride Inorganic materials 0.000 claims description 9
- 239000011259 mixed solution Substances 0.000 claims description 9
- 239000012046 mixed solvent Substances 0.000 claims description 9
- 238000001953 recrystallisation Methods 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 9
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 claims description 8
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 4
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 claims description 4
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical class [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 2
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims description 2
- 238000010792 warming Methods 0.000 claims description 2
- 238000000034 method Methods 0.000 abstract description 12
- 230000015572 biosynthetic process Effects 0.000 abstract description 4
- 230000008569 process Effects 0.000 abstract description 4
- 238000003786 synthesis reaction Methods 0.000 abstract description 4
- 230000006872 improvement Effects 0.000 abstract description 3
- 230000002262 irrigation Effects 0.000 abstract 1
- 238000003973 irrigation Methods 0.000 abstract 1
- 239000000047 product Substances 0.000 description 32
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 15
- 229920006395 saturated elastomer Polymers 0.000 description 14
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 9
- QOSSAOTZNIDXMA-UHFFFAOYSA-N Dicylcohexylcarbodiimide Chemical compound C1CCCCC1N=C=NC1CCCCC1 QOSSAOTZNIDXMA-UHFFFAOYSA-N 0.000 description 8
- 238000005481 NMR spectroscopy Methods 0.000 description 7
- 238000000605 extraction Methods 0.000 description 7
- 239000011780 sodium chloride Substances 0.000 description 7
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 5
- RRONHWAVOYADJL-HNNXBMFYSA-N (2s)-3-phenyl-2-(phenylmethoxycarbonylamino)propanoic acid Chemical compound C([C@@H](C(=O)O)NC(=O)OCC=1C=CC=CC=1)C1=CC=CC=C1 RRONHWAVOYADJL-HNNXBMFYSA-N 0.000 description 4
- 238000005160 1H NMR spectroscopy Methods 0.000 description 4
- 102000004190 Enzymes Human genes 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 4
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 3
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 3
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 238000006555 catalytic reaction Methods 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 3
- 230000006340 racemization Effects 0.000 description 3
- ASOKPJOREAFHNY-UHFFFAOYSA-N 1-Hydroxybenzotriazole Chemical compound C1=CC=C2N(O)N=NC2=C1 ASOKPJOREAFHNY-UHFFFAOYSA-N 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 150000001718 carbodiimides Chemical class 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000006911 enzymatic reaction Methods 0.000 description 2
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 description 2
- 230000020477 pH reduction Effects 0.000 description 2
- 210000000496 pancreas Anatomy 0.000 description 2
- 239000003208 petroleum Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 1
- 102000014413 Neuregulin Human genes 0.000 description 1
- 108050003475 Neuregulin Proteins 0.000 description 1
- TZRXHJWUDPFEEY-UHFFFAOYSA-N Pentaerythritol Tetranitrate Chemical compound [O-][N+](=O)OCC(CO[N+]([O-])=O)(CO[N+]([O-])=O)CO[N+]([O-])=O TZRXHJWUDPFEEY-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241000282894 Sus scrofa domesticus Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 150000008065 acid anhydrides Chemical class 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 150000001263 acyl chlorides Chemical class 0.000 description 1
- 150000008064 anhydrides Chemical class 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 210000000748 cardiovascular system Anatomy 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- RAABOESOVLLHRU-UHFFFAOYSA-N diazene Chemical compound N=N RAABOESOVLLHRU-UHFFFAOYSA-N 0.000 description 1
- 229910000071 diazene Inorganic materials 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000004068 intracellular signaling Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- CUCAZZQPISJXOS-QWRGUYRKSA-N methyl (2s)-2-[[(2s)-2-(phenylmethoxycarbonylamino)propanoyl]amino]propanoate Chemical compound COC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)OCC1=CC=CC=C1 CUCAZZQPISJXOS-QWRGUYRKSA-N 0.000 description 1
- CWVOQXJRSJVARM-HOTGVXAUSA-N methyl (2s)-3-methyl-2-[[(2s)-3-methyl-2-(phenylmethoxycarbonylamino)butanoyl]amino]butanoate Chemical compound COC(=O)[C@H](C(C)C)NC(=O)[C@H](C(C)C)NC(=O)OCC1=CC=CC=C1 CWVOQXJRSJVARM-HOTGVXAUSA-N 0.000 description 1
- SIHNOQQFBTTYLO-YJBOKZPZSA-N methyl (2s)-3-phenyl-2-[[(2s)-2-(phenylmethoxycarbonylamino)propanoyl]amino]propanoate Chemical compound C([C@@H](C(=O)OC)NC(=O)[C@H](C)NC(=O)OCC=1C=CC=CC=1)C1=CC=CC=C1 SIHNOQQFBTTYLO-YJBOKZPZSA-N 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000002858 neurotransmitter agent Substances 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 210000001428 peripheral nervous system Anatomy 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000017854 proteolysis Effects 0.000 description 1
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- 102000005962 receptors Human genes 0.000 description 1
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- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
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Landscapes
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses the application of 1-hydroxyl-1,2,3-phentriazine-4 (3H)-one in Peptide systhesis.Drip irrigation device of the present invention is: 1-hydroxyl-1; 2; the application of 3-phentriazine-4 (3H)-one in Peptide systhesis; mainly through realizing with under type: amino acid methyl ester hydrochloride and carbobenzoxy-(Cbz) protected amino acid are under the condition of organic solvent and organic acid trifluoroacetic acid; by polypeptide condensing agent 1-hydroxyl-1; 2,3-phentriazine-4 (3H)-one and catalyzer 1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate catalyzing and condensing obtain polypeptide.The present invention is that the process of polypeptide condensing agent improvement on synthesis is simple with 1-hydroxyl-1,2,3-phentriazine-4 (3H)-one, is easy to realize, with low cost and the yield of polypeptide is higher.
Description
Technical field
The present invention relates to amino acid polypeptide technical field, be specifically related to the application of 1-hydroxyl-1,2,3-phentriazine-4 (3H)-one in Peptide systhesis.
Background technology
Protein forms one of the most basic material of living organism, and it plays very important effect in the process such as growth, growth, metabolism of cell.Relative to protein, the effect of less polypeptide in organism is also very important.Peptide affects many important biochemical functions in organism, and they participate in receptor-mediated intracellular signaling as neurotransmitter, neuregulins and hormone.There will be a known 100 various active peptides to work in maincenter and peripheral nervous system, cardiovascular systems, immunity system and intestines.In addition, polypeptide class and the medicine based on polypeptide are also the study hotspots of medicine scholars always.The little peptide below 1500 by 2-14 Amino acid profile or molecular weight section, it is worth the highest in all peptides, there is extremely strong biological activity and diversity, there is important biological function, human body physiological function can be played comprehensively, strengthening Human Physiology active, is important physiological regulator ([moral] N. Xiu Ede, H.D. Jia Kubuke work; Liu Keliang, He Junlin etc. translate. " peptide: chemistry and biology " Beijing Science Press, 2005).But isolated peptides is very difficult from natural origin, first, the concentration of peptide is 10 in every milligram of fresh tissue usually
-15-10
-12mo1, it is in intracellular position therefore to only have super-sensitive measuring method just can detect; Secondly, bioactive peptide is obtained generally by its solid vulnerable restriction, as the restriction etc. in tissue source with separation method; Again, collect and store complicated process during organ, as the production of the Pancreas Sus domestica of Regular Insulin or the collection of Pancreas Bovis seu Bubali and storage, further increase the difficulty utilizing natural origin.In addition, if receive the infection of Causative virus for separating of the tissue of bioactive peptide proteolysis matter, then titanic peril can be caused to health.Therefore, the synthesis of polypeptide or polypeptide drug is the problem that must first solve.
The method of Peptide systhesis is divided into enzymatic method and chemical method.As a kind of method forming peptide bond, enzyme' s catalysis has the advantage of its uniqueness.Due to the specificity of enzyme, to amino acid whose side chain functionalities without the need to protection, enzyme' s catalysis is stereospecificity, and the phenomenon of racemization can not occur, and enzyme' s catalysis also has the advantage that can carry out in aqueous.But enzymatic method is also not overripened, not yet reaches the degree of general application, still need further research.At present in the synthesis of chemiluminescent polypeptide method, main employing activated carboxylic method connects reactive polypeptide, using activation of amino acids is the earliest the method for acyl chlorides, nitrine, symmetric anhydride and mixed acid anhydride, but due under these conditions, there is amino acid racemization, reaction reagent is dangerous and prepare the shortcomings such as complicated, replaced by condensation reagent method afterwards gradually.
Polypeptide condensing agent can be divided into diimine type, phosphorus ionic and urea ionic according to its structure.N, N'-dicyclohexylcarbodiimide (DCC) is first the carbodiimide type condensing agent (J.Am.Chem.Soc., 1955,77:1067.) developed in nineteen fifty-five; So far, DCC is still one of condensing agent the most frequently used in Peptide systhesis.But the N that reaction generates, N'-dicyclohexylurea (DCU) (DCU) solubleness in most of organic solvent is very little, mixes being difficult to eliminate in the product sometimes.By immobilized for carbodiimide on superpolymer the condensing agent of gained make the aftertreatment of reaction more simple, such as, the EDC (P-EDC) that resin is immobilized.1966, E.Wiinsch and F.Dress found in DCC condensation reaction, adds N-Hydroxysuccinimide (HOSu) (Chem.Ber.1966,99:110.), substantially increases polypeptide, inhibit racemization and rearrangement reaction.Therefore, on this basis, people attempt further and have developed a series of N-hydroxy derivatives, wherein 1-hydroxyl benzotriazole (HOBt) and 3-hydroxyl-1,2,3-phentriazine-4 (3H)-one (HOOBt) is more conventional, and HOOBt, because it is with low cost and by product is few, becomes desirable polypeptide condensing agent.
Summary of the invention
The object of this invention is to provide the application of a kind of 1-hydroxyl-1,2,3-phentriazine-4 (3H)-one in Peptide systhesis.
Technical scheme of the present invention is: 1-hydroxyl-1, 2, the application of 3-phentriazine-4 (3H)-one in Peptide systhesis, it is characterized in that mainly through realizing with under type: amino acid methyl ester hydrochloride and carbobenzoxy-(Cbz) protected amino acid are under organic solvent and organic acid condition, by polypeptide condensing agent 1-hydroxyl-1, 2, 3-phentriazine-4 (3H)-one (1-HOOBt) and catalyzer 1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDCHCl) catalyzing and condensing obtain polypeptide, described organic solvent is tetrahydrofuran (THF), dimethyl formamide, methyl alcohol, ethanol, propyl alcohol, Virahol, the trimethyl carbinol, ethyl acetate or sherwood oil, be preferably dimethyl formamide, described organic acid is trifluoroacetic acid, described polypeptide condensing agent 1-hydroxyl-1, 2, the structural formula of 3-phentriazine-4 (3H)-one is
.
The application of 1-hydroxyl-1,2,3-phentriazine-4 (3H)-one of the present invention in Peptide systhesis, is characterized in that: the temperature of reaction in Peptide systhesis is-20-50 DEG C.
1-hydroxyl-1 of the present invention, 2, the application of 3-phentriazine-4 (3H)-one in Peptide systhesis, it is characterized in that mainly through realizing with under type: the preparation of (1) amino acid methyl ester hydrochloride, under ice-water bath condition, thionyl chloride is instilled toward being equipped with in methyl alcohol and amino acid whose reaction vessel, after dripping, at room temperature stir 4-5h, concentrating under reduced pressure removes excessive methyl alcohol, repeatedly dissolve also concentrating under reduced pressure with methyl alcohol again and, to remove excessive hydrogenchloride, use dissolve with methanol product, drip ether and separate out white crystal amino acid methyl ester hydrochloride, (2) preparation of carbobenzoxy-(Cbz) protected amino acid, getting amino acid, to be dissolved in volumetric molar concentration be in the sodium hydroxide solution of 2mol/L, and ice-water bath is chilled to 0 DEG C, then Carbobenzoxy Chloride is dripped, dropwise rear continuation and stir 0.5h, be warming up to room temperature, continue stirring reaction 2h, after reaction terminates, reaction solution washed with diethylether twice, then use 6mol/L hydrochloric acid acidizing reaction mixed solution to pH=1-2, be then extracted with ethyl acetate 3 times, merge organic phase, and use anhydrous Na
2sO
4dried overnight, crosses and filters siccative, and underpressure distillation obtains thick product except desolventizing, obtains carbobenzoxy-(Cbz) protected amino acid with ethyl acetate and sherwood oil mixed solvent recrystallization, (3) condensation reaction, the carbobenzoxy-(Cbz) protected amino acid that the amino acid methyl ester hydrochloride obtained by step (1) and step (2) obtain is dissolved in organic solvent, then trifluoroacetic acid is added, by polypeptide condensing agent 1-hydroxyl-1, 2, 3-phentriazine-4 (3H)-one (1-HOOBt) and catalyzer 1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDCHCl) are dissolved in organic solvent respectively, join respectively in reaction solution in the organic solution of the temperature of-20-50 DEG C by the organic solution of polypeptide condensing agent and catalyzer, TLC monitors reaction process, use water successively, 1mol/L hydrochloric acid, saturated sodium carbonate solution, saturated common salt water washing, organic phase anhydrous sodium sulfate drying spends the night, cross and filter siccative, underpressure distillation removing organic solvent obtains thick product, finally carry out column chromatography for separation and obtain white solid polypeptide, described organic solvent is tetrahydrofuran (THF), dimethyl formamide, methyl alcohol, ethanol, propyl alcohol, Virahol, the trimethyl carbinol, ethyl acetate or sherwood oil, be preferably dimethyl formamide.
The application of 1-hydroxyl-1,2,3-phentriazine-4 (3H)-one of the present invention in Peptide systhesis, is characterized in that the principal reaction equation in Peptide systhesis is:
。
The present invention is that the process of polypeptide condensing agent improvement on synthesis is simple with 1-hydroxyl-1,2,3-phentriazine-4 (3H)-one, is easy to realize, with low cost and the yield of polypeptide is higher.
Embodiment
Be described in further details foregoing of the present invention by the following examples, but this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following embodiment, all technology realized based on foregoing of the present invention all belong to scope of the present invention.
Embodiment 1:Z-Phe-Leu-OMe
H
2the preparation of N-Leu-OMeHCl: under ice-water bath condition, 10.4ml (120mmol) thionyl chloride is slowly instilled toward being equipped with in 100ml methyl alcohol and the leucic 250ml round-bottomed flask of 13.2g (100mmol), after dripping, at room temperature stir 4-5h, concentrating under reduced pressure removes excessive methyl alcohol, then with methyl alcohol repeatedly dissolve several times and concentrating under reduced pressure to remove excessive hydrogenchloride, with a small amount of dissolve with methanol product, drip ether and separate out white crystal 17.2g, yield 95%.
The preparation of Cbz-Phe-OH: get 19.8g (120mmol) phenylalanine and be dissolved in the NaOH solution of 180ml2mol/L, ice-water bath is chilled to 0 DEG C; Then slowly drip 10.2g (60mmol) Carbobenzoxy Chloride, dropwise rear continuation and stir 0.5h; Then slowly rise to room temperature, continue stirring reaction 2h; After reaction terminates, reaction solution 20ml washed with diethylether twice, and then with 6mol/L hydrochloric acid acidizing reaction mixed solution to pH=1-2, then use 30ml extraction into ethyl acetate 3 times; Merge organic phase, and use anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; White crystal 16.3g is obtained, yield 91% with ethyl acetate and sherwood oil mixed solvent recrystallization.
Condensation reaction: the H getting 2.993g (10mmol) Cbz-Phe-OH, 2.180g (12mmol)
2n-Leu-OMeHCI is placed in 250ml round-bottomed flask, add 50ml dimethyl formamide to dissolve, add 36mmol trifluoroacetic acid, the EDCHCl getting 3mmol is dissolved in 10ml dimethyl formamide, under room temperature, the 40ml dimethyl formamide solution being dissolved with 12mmol1-HOOBt is slowly added above-mentioned solution.
TLC monitors reaction process, uses water (300ml), 1mol/L hydrochloric acid (300ml), saturated Na successively
2cO
3(300ml), saturated aqueous common salt (300ml) washing; Organic phase anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; Finally obtain white solid 4.125g, productive rate 92%, purity 95% with carrying out column chromatography for separation.
Target product characterizes:
1H-NMR(400MHz,CDCl
3):0.88(t,6H),1.41-1.60(m,3H),3.02-3.13(m,2H),3.69(s,3H),4.47(d,1H),4.53-4.58(m,1H),5.08(s,2H),5.39(brd,1H),6.30(brd,1H),7.17-7.32(m,10H).
13C-NMR(100MHz,CDCl
3):172.8,170.5,155.9,136.2,136.0,129.3,128.6,128.5,128.2,128.0,127.0,67.0,56.0,52.2,50.7,41.4,38.3,24.6,22.6,21.8;MS(ESI,m/z):449.2[M+Na]
+.
Embodiment 2:Z-Val-Val-OMe
H
2the preparation of N-Val-OMeHCl: under ice-water bath condition, 5.2ml (60mmol) thionyl chloride is slowly instilled in the 100ml round-bottomed flask that 50ml methyl alcohol and 5.9g (50mmol) α-amino-isovaleric acid be housed, after dripping, at room temperature stir 4-5h, concentrating under reduced pressure removes excessive methyl alcohol, then with methyl alcohol repeatedly dissolve several times and concentrating under reduced pressure to remove excessive hydrogenchloride, with a small amount of dissolve with methanol product, drip ether and separate out white crystal 7.9g, yield 94%.
The preparation of Cbz-Val-OH: get 7.0g (60mmol) α-amino-isovaleric acid and be dissolved in the NaOH solution of 90ml2mol/L, ice-water bath is chilled to 0 DEG C; Then slowly drip 5.1g (30mmol) Carbobenzoxy Chloride, dropwise rear continuation and stir 0.5h; Then slowly rise to room temperature, continue stirring reaction 2h; After reaction terminates, reaction solution 10ml washed with diethylether twice, and then with 6mol/L hydrochloric acid acidizing reaction mixed solution to pH=1-2, then use 15ml extraction into ethyl acetate 3 times; Merge organic phase, and use anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; By acetic acid second, cruel and sherwood oil mixed solvent recrystallization obtains white crystal 6.9g, yield 92%.
Condensation reaction: the H getting 1.257g (5mmol) Cbz-Val-OH, 1.006g (6mmol)
2n-Val-OMeHCI is placed in 100ml round-bottomed flask, add 25ml dimethyl formamide to dissolve, add 18mmol trifluoroacetic acid, the EDCHCl getting 1.5mmol is dissolved in 5ml dimethyl formamide, under room temperature, the 20ml dimethyl formamide solution being dissolved with 6mmol1-HOOBt is slowly added above-mentioned solution.
TLC monitors reaction process, uses water (150ml), 1mol/L hydrochloric acid (150ml), saturated Na successively
2cO
3(150ml), saturated aqueous common salt (150ml) washing; Organic phase anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; Finally obtain white solid 1.822g, yield 94%, purity 96% with carrying out column chromatography for separation.
Target product characterizes:
1H-NMR(400MHz,CDCl
3):0.88-0.98(m,12H),2.06-2.21(m,2H),3.73(s,3H),4.10(dd,1H),4.55(dd,1H),5.12(s,2H),5.47(brd,1H),6.55(brd,1H),7.31(m,5H).
13C-NMR(100MHz,CDCl
3):172.2,171.3,156.4,136.2,128.5,128.1，128.0,67.0,60.3,57.1，52.1,31.1,19.1,18.9,17.8,17.7;MS(ESI,m/z):387.2[M+Na]
+。
Embodiment 3:Z-Ala-Ala-OMe
H
2the preparation of N-Ala-OMeHCI: under ice-water bath condition, 6.2ml (72mmol) thionyl chloride is slowly instilled in the 120ml round-bottomed flask that 60ml methyl alcohol and 5.4g (60mmol) L-Ala be housed, after dripping, at room temperature stir 4-5h, concentrating under reduced pressure removes excessive methyl alcohol, then with methyl alcohol repeatedly dissolve several times and concentrating under reduced pressure to remove excessive hydrogenchloride, with a small amount of dissolve with methanol product, drip ether and separate out white crystal 7.5g, yield 90%.
The preparation of Cbz-Ala-OH: get 6.4g (72mmol) L-Ala and be dissolved in the NaOH solution of 108ml2mol/L, ice-water bath is chilled to 0 DEG C; Then slowly drip 6.1g (36mmol) Carbobenzoxy Chloride, dropwise rear continuation and stir 0.5h; Then slowly rise to room temperature, continue stirring reaction 2h; After reaction terminates, reaction solution 10ml washed with diethylether twice, and then with 6mol/L hydrochloric acid acidizing reaction mixed solution to pH=1-2, then use 18ml extraction into ethyl acetate 3 times; Merge organic phase, and use anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; By acetic acid second, cruel and sherwood oil mixed solvent recrystallization obtains white crystal 7.31g, yield 91%.
Condensation reaction: the H getting 446.4mg (2mmol) Cbz-Ala-OH, 335.0mg (2.4mmol)
2n-Ala-OMeHCI is placed in 50ml round-bottomed flask, add 10ml dimethyl formamide to dissolve, add 7.2mmol trifluoroacetic acid, the EDCHCl getting 0.6mmol is dissolved in 2ml dimethyl formamide, under room temperature, the 8ml dimethyl formamide solution being dissolved with 2.4mmol1-HOOBt is slowly added above-mentioned solution.
TLC monitors reaction process, uses water (60ml), 1mol/L hydrochloric acid (60ml), saturated Na successively
2cO
3(60ml), saturated aqueous common salt (60ml) washing; Organic phase anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; Finally obtain white solid 0.62g, productive rate 94%, purity 95% with carrying out column chromatography for separation.
Target product characterizes:
1h-NMR (400MHz, CDCl
3): 1.38 (d, 6H), 3.74 (s, 3H), 4.31 (t, 1H); 4.56 (p, 1H), 5.10 (s, 2H), 5.56 (brd; 1H), 6.79 (brd, 1H), 7.32 (m, 5H);
13c-NMR (100MHz, CDCl
3): 173.1,171.9,155.9,136.1,128.5,128.1,128.0,66.9,52.4,50.3,48.0,18.7,18.1; MS (ESI, m/z): 331.2 (M+Na]
ten.
Embodiment 4:Z-Ala-Phe-OMe
H
2the preparation of N-Phe-OMeHCI: under ice-water bath condition, 5.2ml (60mmol) thionyl chloride is slowly instilled in the 100ml round-bottomed flask that 50ml methyl alcohol and 8.3g (50mmol) phenylalanine be housed, after dripping, at room temperature stir 4-5h, concentrating under reduced pressure removes excessive methyl alcohol, then with methyl alcohol repeatedly dissolve several times and concentrating under reduced pressure to remove excessive hydrogenchloride, with a small amount of dissolve with methanol product, drip ether and separate out white crystal 10.0g, yield 93%.
The preparation of Cbz-Ala-OH: get 6.4g (72mmol) L-Ala and be dissolved in the NaOH solution of 108ml2mol/L, ice-water bath is chilled to 0 DEG C; Then slowly drip 6.1g (36mmol) Carbobenzoxy Chloride, dropwise rear continuation and stir 0.5h; Then slowly rise to room temperature, continue stirring reaction 2h; After reaction terminates, reaction solution 10ml washed with diethylether twice, and then with 6mol/L hydrochloric acid acidizing reaction mixed solution to pH=1-2, then use 18ml extraction into ethyl acetate 3 times; Merge organic phase, and use anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; By acetic acid second, cruel and sherwood oil mixed solvent recrystallization obtains white crystal 7.31g, yield 91%.
Condensation reaction: the H getting 1.111g (5mmol) Cbz-Ala-OH, 837.5mg (6mmol)
2n-Phe-OMeHCI is placed in 100ml round-bottomed flask, add 25ml tetrahydrofuran (THF) to dissolve, add 18mmol trifluoroacetic acid, the EDCHCl getting 1.5mmol is dissolved in 5ml tetrahydrofuran (THF), under room temperature, the 20ml tetrahydrofuran solution being dissolved with 6mmol1-HOOBt is slowly added above-mentioned solution.
TLC monitors reaction process, uses water (150ml), 1mol/L hydrochloric acid (150ml), saturated Na successively
2cO
3(150ml), saturated aqueous common salt (150ml) washing; Organic phase anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; Finally obtain white solid 1.873g, yield 92%, purity 95% with carrying out column chromatography for separation.
Target product characterizes:
1H-NMR(400MHz,CDCl
3:1.32(d,3H),3.06(dd,1H),3.14(dd,1H),3.71(s,3H),4.24(t,3H),4.85(dd,1H),5.08(dd,2H),5.34(brd,1H),6.56(brd,1H),7.08(d,2H),7.20-7.30(m,8H);
13c-NMR (100MHz, CDCl
3): 171.8,171.6,155.8,136.1,135.6,129.2,128.6,128.5,128.2,128.0,127.1,67.0,53.1,52.3,50.3,37.7,18.4; MS (ESI): 407.2 [M+Na]
ten.
Embodiment 5:Z-Ala-Val-OMe
H
2the preparation of N-Val-OMeHCl: under ice-water bath condition, 5.2ml (60mmol) thionyl chloride is slowly instilled in the 100ml round-bottomed flask that 50ml methyl alcohol and 5.9g (50mmol) α-amino-isovaleric acid be housed, after dripping, at room temperature stir 4-5h, concentrating under reduced pressure removes excessive methyl alcohol, then with methyl alcohol repeatedly dissolve several times and concentrating under reduced pressure to remove excessive hydrogenchloride, with a small amount of dissolve with methanol product, drip ether and separate out white crystal 7.9g, yield 94%.
The preparation of Cbz-Ala-OH: get 6.4g (72mmol) L-Ala and be dissolved in the NaOH solution of 108ml2mol/L, ice-water bath is chilled to 0 DEG C; Then slowly drip 6.1g (36mmol) Carbobenzoxy Chloride, dropwise rear continuation and stir 0.5h; Then slowly rise to room temperature, continue stirring reaction 2h, after reaction terminates, reaction solution 10ml washed with diethylether twice, and then with 6mol/L hydrochloric acid acidizing reaction mixed solution to pH=1-2, then use 18ml extraction into ethyl acetate 3 times; Merge organic phase, and use anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; By acetic acid second, cruel and sherwood oil mixed solvent recrystallization obtains white crystal 7.31g, yield 91%.
Condensation reaction: the H getting 222.2mg (1mmol) Cbz-Ala-OH, 201.2mg (1.2mmol)
2n-Val-OMeHCI is placed in 25ml round-bottomed flask, adds 5ml dissolve with ethanol, adds 3.6mmol trifluoroacetic acid, and the EDCHCl getting 0.3mmol is dissolved in 1ml ethanol, under room temperature, the 4ml ethanolic soln being dissolved with 1.2mmol1-HOOBt is slowly added above-mentioned solution.
TLC monitors reaction process, uses water (30ml), 1mol/L hydrochloric acid (30mL), saturated Na successively
2cO
3(30ml), saturated aqueous common salt (30ml) washing; Organic phase anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; Finally obtain white solid 0.3412g, yield 95%, purity 96% with carrying out column chromatography for separation.
Target product characterizes:
1h-NMR (400MHz, CDCl
3): 0.89 (dd, 6H), 1.38 (d, 3H); 2.11-2.19 (m, 1H), 3.73 (s, 3H); 4.33 (t, 1H), 4.53 (dd; 1H), 5.11 (s, 2H); 5.51 (brd, 1H), 6.69 (brd; 1H), 7.34 (m, 5H);
13c-NMR (100MHz, CDCl
3): 172.2,172.1,155.9,136.1,128.5,128.1,128.0,66.9,57.1,52.1,50.4,31.1,18.8,18.3,17.6; MS (ESI): 359.2 [M+Na]
ten.
Embodiment 6:Z-Ala-Gly-OEt
H
2the preparation of N-Gly-OEtHCI: under ice-water bath condition, 6.2ml (72mmol) thionyl chloride is slowly instilled in the 100ml round-bottomed flask that 60ml methyl alcohol and 4.5g (60mmol) glycine be housed, after dripping, at room temperature stir 4-5h, concentrating under reduced pressure removes excessive methyl alcohol, then with methyl alcohol repeatedly dissolve several times and concentrating under reduced pressure to remove excessive hydrogenchloride, with a small amount of dissolve with methanol product, drip ether and separate out white crystal 7.6g, yield 91%.
The preparation of Cbz-Ala-OH: get 6.4g (72mmol) L-Ala and be dissolved in the NaOH solution of 108ml2mol/L, ice-water bath is chilled to 0 DEG C; Then slowly drip 6.1g (36mmol) Carbobenzoxy Chloride, dropwise rear continuation and stir 0.5h; Then slowly rise to room temperature, continue stirring reaction 2h; After reaction terminates, reaction solution 10ml washed with diethylether twice, and then with 6mol/LHCl acidification reaction mixed solution to pH=1-2, then use 18ml extraction into ethyl acetate 3 times; Merge organic phase, and use anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; By acetic acid second, cruel and sherwood oil mixed solvent recrystallization obtains white crystal 7.31g, yield 91%.
Condensation reaction: the H getting 1.111g (5mmol) Cbz-Ala-OH, 837.5mg (6mmol)
2n-Gly-OEtHCI is placed in 100ml round-bottomed flask, add 25ml acetic acid ethyl dissolution, add 18mmol trifluoroacetic acid, the EDCHCl getting 1.5mmol is dissolved in 5ml ethyl acetate, in the temperature of-25 DEG C, the 20ml ethyl acetate solution being dissolved with 6mmol1-HOOBt is slowly added above-mentioned solution.
TLC monitors reaction process, uses water (150ml), 1mol/L hydrochloric acid (150ml), saturated Na successively
2cO
3(150ml), saturated aqueous common salt (150ml) washing; Organic phase anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; Finally obtain white solid 1.537g, yield 93%, purity 94% with carrying out column chromatography for separation.
Target product characterizes:
1H-NMR(400MHz,CDCl
3):1.27(t,3H),1.39(d,3H),4.00(d,2H),4.19(q,2H),4.33(t,1H),5.10(dd,2H),5.59(brd,1H),6.83(brs,1H),7.33(m,5H);
13C-NMR(100MHz,CDCl
3):172.6,169.7,156.0,136.1,128.5,128.1,128.0,67.0,61.5,50.3,41.2,18.5,14.0;MS(ESI):331.2[M+Na]
+。
Embodiment 7:Z-Phe-Gly-OEt
H
2the preparation of N-Gly-OEtHCI: under ice-water bath condition, 6.2ml (72mmol) thionyl chloride is slowly instilled in the 100ml round-bottomed flask that 60ml methyl alcohol and 4.5g (60mmol) glycine be housed, after dripping, at room temperature stir 4-5h, concentrating under reduced pressure removes excessive methyl alcohol, then with methyl alcohol repeatedly dissolve several times and concentrating under reduced pressure to remove excessive hydrogenchloride, with a small amount of dissolve with methanol product, drip ether and separate out white crystal 7.6g, yield 91%.
The preparation of Cbz-Phe-OH: get 19.8g (120mmol) phenylalanine and be dissolved in the NaOH solution of 180ml2mol/L, ice-water bath is chilled to 0 DEG C; Then slowly drip 10.2g (60mmol) Carbobenzoxy Chloride, dropwise rear continuation and stir 0.5h; Then slowly rise to room temperature, continue stirring reaction 2h; After reaction terminates, reaction solution 20ml washed with diethylether twice, and then with 6mol/LHCl acidification reaction mixed solution to pH=1-2, then use 30ml extraction into ethyl acetate 3 times; Merge organic phase, and use anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; White crystal 16.3g is obtained, yield 91% with ethyl acetate and sherwood oil mixed solvent recrystallization.
Condensation reaction: the H getting 299.3mg (1mmol) Cbz-Phe-OH, 167.5mg (1.2mmol)
2n-Gly-OEtHCI is placed in 25ml round-bottomed flask, add 5ml petroleum ether dissolution, add 3.6mmol trifluoroacetic acid, the EDCHCl getting 0.3mmol is dissolved in 1ml sherwood oil, in the temperature of 50 DEG C, the 4ml petroleum ether solution being dissolved with 1.2mmol1-HOOBt is slowly added above-mentioned solution.
TLC monitors reaction process, uses water (30ml), 1mol/L hydrochloric acid (30ml), saturated Na successively
2cO
3(30ml), saturated aqueous common salt (30ml) washing; Organic phase anhydrous Na
2sO
4dried overnight; Cross and filter siccative, underpressure distillation removes desolventizing and obtains thick product; Finally obtain white solid 0.387g, yield 95%, purity 94% with carrying out column chromatography for separation.
Target product characterizes:
1h-NMR (400MHz, CDCl
3): 1.25 (t, 3H), 3.03-3.14 (m, 2H); 3.89 (dd, 1H), 3.99 (dd, 1H); 4.17 (q, 2H), 4.50 (d; 1H), 5.05 (dd, 2H); 5.48 (brd, 1H), 6.52 (brs; 1H), 7.17-7.30 (m, 10H);
13c-NMR (100MHz, CDCl
3): 171.2,169.4,156.0,136.3,129.2,128.6,128.5,128.1,127.9,127.0,67.0,61.5,56.0,41.3,38.4,14.1; MS (ESI): 407.2 [M+Na]
ten.
Embodiment above describes ultimate principle of the present invention, principal character and advantage.The technician of the industry should understand; the present invention is not restricted to the described embodiments; what describe in above-described embodiment and specification sheets just illustrates principle of the present invention; under the scope not departing from the principle of the invention; the present invention also has various changes and modifications, and these changes and improvements all fall in the scope of protection of the invention.
Claims (5)
1.1-hydroxyl-1, 2, the application of 3-phentriazine-4 (3H)-one in Peptide systhesis, it is characterized in that mainly through realizing with under type: amino acid methyl ester hydrochloride and carbobenzoxy-(Cbz) protected amino acid are under organic solvent and organic acid condition, by polypeptide condensing agent 1-hydroxyl-1, 2, 3-phentriazine-4 (3H)-one and catalyzer 1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate catalyzing and condensing obtain polypeptide, described organic solvent is tetrahydrofuran (THF), dimethyl formamide, methyl alcohol, ethanol, propyl alcohol, Virahol, the trimethyl carbinol, ethyl acetate or sherwood oil, described organic acid is trifluoroacetic acid, described polypeptide condensing agent 1-hydroxyl-1, 2, the structural formula of 3-phentriazine-4 (3H)-one is
.
2. the application of 1-hydroxyl-1,2,3-phentriazine-4 (3H)-one according to claim 1 in Peptide systhesis, is characterized in that: the temperature of reaction in Peptide systhesis is-20-50 DEG C.
3. 1-hydroxyl-1 according to claim 1,2, the application of 3-phentriazine-4 (3H)-one in Peptide systhesis, it is characterized in that mainly through realizing with under type: the preparation of (1) amino acid methyl ester hydrochloride, under ice-water bath condition, thionyl chloride is instilled toward being equipped with in methyl alcohol and amino acid whose reaction vessel, after dripping, at room temperature stir 4-5h, concentrating under reduced pressure removes excessive methyl alcohol, repeatedly dissolve also concentrating under reduced pressure with methyl alcohol again and, to remove excessive hydrogenchloride, use dissolve with methanol product, drip ether and separate out white crystal amino acid methyl ester hydrochloride, (2) preparation of carbobenzoxy-(Cbz) protected amino acid, getting amino acid, to be dissolved in volumetric molar concentration be in the sodium hydroxide solution of 2mol/L, and ice-water bath is chilled to 0 DEG C, then Carbobenzoxy Chloride is dripped, dropwise rear continuation and stir 0.5h, be warming up to room temperature, continue stirring reaction 2h, after reaction terminates, reaction solution washed with diethylether twice, then use 6mol/L hydrochloric acid acidizing reaction mixed solution to pH=1-2, be then extracted with ethyl acetate 3 times, merge organic phase, and use anhydrous Na
2sO
4dried overnight, crosses and filters siccative, and underpressure distillation obtains thick product except desolventizing, obtains carbobenzoxy-(Cbz) protected amino acid with ethyl acetate and sherwood oil mixed solvent recrystallization, (3) condensation reaction, the carbobenzoxy-(Cbz) protected amino acid that the amino acid methyl ester hydrochloride obtained by step (1) and step (2) obtain is dissolved in organic solvent, then trifluoroacetic acid is added, by polypeptide condensing agent 1-hydroxyl-1, 2, 3-phentriazine-4 (3H)-one and catalyzer 1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate are dissolved in organic solvent respectively, join respectively in reaction solution in the organic solution of the temperature of-20-50 DEG C by the organic solution of polypeptide condensing agent and catalyzer, TLC monitors reaction process, use water successively, 1mol/L hydrochloric acid, saturated sodium carbonate solution, saturated common salt water washing, organic phase anhydrous sodium sulfate drying spends the night, cross and filter siccative, underpressure distillation removing organic solvent obtains thick product, finally carry out column chromatography for separation and obtain white solid polypeptide, described organic solvent is tetrahydrofuran (THF), dimethyl formamide, methyl alcohol, ethanol, propyl alcohol, Virahol, the trimethyl carbinol, ethyl acetate or sherwood oil.
4. the application of 1-hydroxyl-1,2,3-phentriazine-4 (3H)-one in Peptide systhesis according to claim 1 or 3, is characterized in that: described organic solvent is dimethyl formamide.
5. the application of 1-hydroxyl-1,2,3-phentriazine-4 (3H)-one in Peptide systhesis according to claim 1 or 3, is characterized in that the principal reaction equation in Peptide systhesis is:
。
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