CN101525368B - Deprotection agent for synthesizing polypeptide - Google Patents
Deprotection agent for synthesizing polypeptide Download PDFInfo
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- CN101525368B CN101525368B CN200810085323XA CN200810085323A CN101525368B CN 101525368 B CN101525368 B CN 101525368B CN 200810085323X A CN200810085323X A CN 200810085323XA CN 200810085323 A CN200810085323 A CN 200810085323A CN 101525368 B CN101525368 B CN 101525368B
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- polypeptide
- deprotection
- deprotection agent
- dbu
- hobt
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Abstract
The invention relates to a deprotection agent for synthesizing polypeptide solid phase, which is applied to amino-group deprotection in polypeptide Fmoc solid phase synthesis. The formulation of the deprotection agent comprises 0.1 to 10 percent of piperidine, 0.1 to 5 percent of DBU, 0.5 to 5 percent of HOBt, 0.1 to 2 percent of Tritionx-100, and DMF as a solvent. The deprotection agent is added with reasonable proportions of the piperidine, the HOBt, the DBU and the Tritionx-100 so that amino groups in polypeptide synthesis can be deprotected and bonded with amino acid efficiently, the synthesized polypeptide has high purity, an organic reagent used in the deprotection process is omitted, and the cost of synthesizing the polypeptide is reduced.
Description
Technical field
It is synthetic that the present invention relates to polypeptide, particularly the amino deprotection agent in the polypeptide Fmoc solid phase synthesis.
Background technology
Merrifield had successfully synthesized polypeptide in 1963, so the solid phase synthesis of polypeptide develops gradually.Polypeptide is synthetic to be one and to repeat to add amino acid whose process, and the solid phase synthesis order is generally synthetic to N end (aminoterminal) from C end (carboxyl terminal).The amino acid that to form the purpose peptide earlier is transformed into the amino acid that N-terminal has the protection base, and amino acid all is the protection of N-a-amino acid in Boc and Fmoc synthesis method.Carboxyl terminal is a free, and aminoterminal must first deprotection and carboxyl terminal activation, condensation spreading peptide chain then before reaction.Solid phase synthesis process has two kinds, i.e. Fmoc and Boc.The Boc synthesis method is with TFA acid deprotection, but inapplicable contain tryptophane etc. to the synthetic of the unsettled peptide class of acid because acid can make peptide come off from resin, loss is serious, and s.t. also can cause the side reaction of side chain.The Fmoc method overcomes above shortcoming with weakly alkaline piperidines/DMF=1: 4 are deprotection agent; The protection but this deprotection agent can not deaminize well, influence is the condensation reaction in step down, and synthetic polypeptide speed is low, purity is not high; And the organic solvent amount that consumes is also very big, and production cost is high.
Summary of the invention
The shortcoming that the polypeptide production cost is high, purity is low that incomplete deprotection causes with producing side reaction when synthetic for fear of polypeptide.The present invention is through to the deprotection agent Formula Design, improves the amino efficient of going to protect, the polypeptide of synthesis of high purity, and practiced thrift production cost.
Technical scheme of the present invention is
Prescription: piperidines 0.1%-10%
DBU 0.1％-5％
HOBt 0.5％-5％
Trition?x-100 0.1％-2％
Wherein solvent is DMF, and piperidines, DBU, Trition x-100 are the volume ratio components, and HOBt is a weight ratio ingredient.
Through in the deprotection agent piperidines/DMF of routine, adding acvator HOBt and tensio-active agent Trition x-100, catalyzer DBU.HOBt is an acvator, can make that reaction is fast when synthetic, racemization is few, side reaction is few; Trition x-100 tensio-active agent can make the polypeptide spread apart, makes deprotection connect next amino acid fully and efficiently; DBU is soft alkali as catalyzer, be prone to remove amino protecting group, and piperidines more impels amino Fmoc blocking group to remove having in the presence of the DBU.
Select the proper ratio scope of above prescription for use, saved consumptions such as using the organic reagent piperidines in the deprotection, also remove amido protecting efficiently, and easy condensation, thereby building-up reactions is accelerated, the polypeptide of synthesis of high purity.
The deprotection agent of this prescription proves that its effect is obvious than other deprotection agent, synthetic polypeptide purity height through in the test of multiple polypeptides solid phase synthesis repeatedly.Wherein the most adaptive ratio of piperidines is 0.5%-7%, and the most adaptive ratio of DBU is 0.5%-3%, and the most adaptive ratio of HOBt is 1%-2%, and the most adaptive ratio of Trition x-100 is 0.5%-1%.
Benefit of the present invention is that this deprotection agent can remove amido protecting efficiently, promotes condensation reaction.The polypeptide resultant velocity is accelerated, and purity is high, has reduced the usage quantity of synthetic middle organic reagent, has reduced polypeptide synthetic cost.
Embodiment
Embodiment 1
Prescription: piperidines 0.5%, DBU 0.5%, and HOBt 1%, Trition x-100 0.5%, surplus is DMF.
At thymus gland α
1Use this prescription to remove amido protecting in synthetic, through detecting, synthetic thick peptide purity is greater than 75%.Under identical experiment condition, use traditional deprotection agent, synthetic thick peptide purity is less than 50%.
Embodiment 2
Prescription: piperidines 0.1%, DBU 0.1%, and HOBt 0.5%, Trition x-100 0.1%, surplus is DMF.
At thymus gland α
1Use this prescription to remove amido protecting in synthetic, through detecting, final thick peptide purity is greater than 70%.
Under identical experiment condition, use traditional deprotection agent, final thick peptide purity is less than 50%.
Embodiment 3
Prescription: piperidinyl-1 0%, DBU 5%, and HOBt 5%, Trition x-100 2%, surplus is DMF.
At thymus gland α
1Remove amido protecting with this prescription in synthetic, through detecting, synthetic thick peptide purity under identical experiment condition, is used traditional deprotection agent greater than 70%, and synthetic thick peptide purity is less than 50%.
Claims (5)
1. deprotection agent for synthesizing polypeptide; The proportioning that it is characterized in that each component: piperidines 0.1%-10%, DBU 0.1%-5%, HOBt 0.5%-5%, Trition x-100 0.1%-2%; Wherein solvent is DMF; Piperidines, DBU, Trition x-100 are the volume ratio components, and HOBt is a weight ratio ingredient.
2. deprotection agent according to claim 1 is characterized in that the most adaptive ratio of piperidines is 0.5%-7%.
3. deprotection agent according to claim 1 is characterized in that the most adaptive ratio of DBU is 0.5%-3%.
4. deprotection agent according to claim 1 is characterized in that the most adaptive ratio of HOBt is 1%-2%.
5. deprotection agent according to claim 1 is characterized in that the most adaptive ratio of Tritionx-100 is 0.5%-1%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810085323XA CN101525368B (en) | 2008-03-06 | 2008-03-06 | Deprotection agent for synthesizing polypeptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810085323XA CN101525368B (en) | 2008-03-06 | 2008-03-06 | Deprotection agent for synthesizing polypeptide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101525368A CN101525368A (en) | 2009-09-09 |
| CN101525368B true CN101525368B (en) | 2012-05-30 |
Family
ID=41093461
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200810085323XA Active CN101525368B (en) | 2008-03-06 | 2008-03-06 | Deprotection agent for synthesizing polypeptide |
Country Status (1)
| Country | Link |
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| CN (1) | CN101525368B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101906150B (en) * | 2010-06-28 | 2013-01-09 | 上海昂博生物技术有限公司 | Preparation method of Bivalirudin |
| MX365465B (en) | 2013-03-21 | 2019-06-04 | Sanofi Aventis Deutschland | Synthesis of cyclic imide containing peptide products. |
| HUE034308T2 (en) | 2013-03-21 | 2018-02-28 | Sanofi Aventis Deutschland | Synthesis of hydantoin containing peptide products |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1865283A (en) * | 2005-05-17 | 2006-11-22 | 周达明 | Solid phase polypeptide synthesis preparation method for salcatonin |
-
2008
- 2008-03-06 CN CN200810085323XA patent/CN101525368B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1865283A (en) * | 2005-05-17 | 2006-11-22 | 周达明 | Solid phase polypeptide synthesis preparation method for salcatonin |
Non-Patent Citations (7)
| Title |
|---|
| ANDREW B. CLIPPINGDALE等.Peptide Thioester Preparation by Fmoc Solid Phase Peptide Synthesis for Use in Native Chemical Ligation.《Journal of Peptide Science》.2000 European Peptide Society and John Wiley & Sons, Ltd.,2000,第6卷第225－234页. * |
| Béla Tö |
| Béla Török等.Deprotection and cleavage of peptides bound to Merrifield resin by stable dimethyl ether–poly(hydrogen fluoride) (DMEPHF) complex. a new and convenient reagent for peptide chemistry.《CHEM. COMMUN.》.The Royal Society of Chemistry 2002,2002,第2882-2883页. * |
| k等.Deprotection and cleavage of peptides bound to Merrifield resin by stable dimethyl ether–poly(hydrogen fluoride) (DMEPHF) complex. a new and convenient reagent for peptide chemistry.《CHEM. COMMUN.》.The Royal Society of Chemistry 2002,2002,第2882－2883页. |
| rö |
| Xianzhang Bu等.An improved deblocking agent for direct Fmoc solid-phase synthesis of peptide thioesters.《Tetrahedron Letters》.2002 Elsevier Science Ltd.,2002,第43卷第2419-2422页. * |
| 韩香等.固相法在多肽合成领域的应用.《药学进展》.2004,第28卷(第1期),第10-14页. * |
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| Publication number | Publication date |
|---|---|
| CN101525368A (en) | 2009-09-09 |
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Effective date of registration: 20191204 Address after: The Town Tower Ridge District Xifeng road 571200 in Hainan Province, Ding'an County Patentee after: HAINAN JIANKE PHARMACEUTICAL CO.,LTD. Address before: 570125 financial garden C-903, North Road, Hainan, Haikou, China World Trade Center Patentee before: HAINAN JIANBANG PHARMACEUTICAL TECHNOLOGY CO.,LTD. |
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Effective date of registration: 20210301 Address after: 570100 room 13a01, Chengxi business center, 146 Longkun South Road, Longhua District, Haikou City, Hainan Province Patentee after: HAINAN JIANBANG PHARMACEUTICAL TECHNOLOGY Co.,Ltd. Address before: 571200 Qifeng Road, Taling New Area, Dingcheng Town, Dingan County, Hainan Province Patentee before: HAINAN JIANKE PHARMACEUTICAL Co.,Ltd. |
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Address after: Room 308, 3rd Floor, Chuangye Building, 168 Nanhai Avenue, Haikou Free Trade Zone, Hainan Province, 570100 Patentee after: HAINAN JIANBANG PHARMACEUTICAL TECHNOLOGY CO.,LTD. Address before: 570100 room 13a01, Chengxi business center, 146 Longkun South Road, Longhua District, Haikou City, Hainan Province Patentee before: HAINAN JIANBANG PHARMACEUTICAL TECHNOLOGY CO.,LTD. |