CN101525368B - Deprotection agent for polypeptide synthesis - Google Patents
Deprotection agent for polypeptide synthesis Download PDFInfo
- Publication number
- 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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- CN
- China
- Prior art keywords
- polypeptide
- deprotection
- deprotection agent
- dbu
- hobt
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 34
- 238000010511 deprotection reaction Methods 0.000 title claims abstract description 28
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 24
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 24
- 230000015572 biosynthetic process Effects 0.000 title abstract description 6
- 238000003786 synthesis reaction Methods 0.000 title abstract description 6
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 20
- NPZTUJOABDZTLV-UHFFFAOYSA-N hydroxybenzotriazole Substances O=C1C=CC=C2NNN=C12 NPZTUJOABDZTLV-UHFFFAOYSA-N 0.000 claims abstract description 13
- 239000002904 solvent Substances 0.000 claims abstract description 3
- 150000003053 piperidines Chemical class 0.000 claims description 12
- 230000003044 adaptive effect Effects 0.000 claims description 8
- 239000004615 ingredient Substances 0.000 claims description 2
- 230000002194 synthesizing effect Effects 0.000 claims description 2
- 238000010532 solid phase synthesis reaction Methods 0.000 abstract description 7
- 150000001413 amino acids Chemical class 0.000 abstract description 6
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 abstract description 5
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 abstract description 4
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 3
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 abstract 4
- 125000003277 amino group Chemical group 0.000 abstract 1
- 125000003368 amide group Chemical group 0.000 description 5
- 239000002253 acid Substances 0.000 description 3
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 238000007086 side reaction Methods 0.000 description 3
- 210000001541 thymus gland Anatomy 0.000 description 3
- 239000013543 active substance Substances 0.000 description 2
- 238000009833 condensation Methods 0.000 description 2
- 230000005494 condensation Effects 0.000 description 2
- 238000006482 condensation reaction Methods 0.000 description 2
- 238000001308 synthesis method Methods 0.000 description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 239000006035 Tryptophane Substances 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 125000006239 protecting group Chemical group 0.000 description 1
- 230000006340 racemization Effects 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 229960004799 tryptophan Drugs 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a deprotection agent for polypeptide synthesis, and relates to a deprotection agent for polypeptide solid phase synthesis, which is used for amino deprotection in polypeptide Fmoc solid phase synthesis. The formula comprises the following components: piperidine 0.1-10%, DBU 0.1-5%, HOBt 0.5-5%, Trition x-1000.1-2%, and DMF as solvent. The piperidine, HOBt, DBU and Trition x-100 which are reasonably proportioned are added into the deprotection agent, so that amino groups can be efficiently deprotected and amino acids can be grafted during polypeptide synthesis, the synthesized polypeptide has high purity, organic reagents used in deprotection are saved, and the polypeptide synthesis cost 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 polypeptide synthesis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200810085323XA CN101525368B (en) | 2008-03-06 | 2008-03-06 | Deprotection agent for polypeptide synthesis |
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 polypeptide synthesis |
Country Status (1)
| Country | Link |
|---|---|
| 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 |
| HUE034308T2 (en) | 2013-03-21 | 2018-02-28 | Sanofi Aventis Deutschland | Synthesis of hydantoin containing peptide products |
| CN105102427B (en) | 2013-03-21 | 2018-09-07 | 赛诺菲-安万特德国有限公司 | The synthesis of peptide prod containing cyclic imide |
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 (4)
| 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 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. * |
| 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. * |
Also Published As
| 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. |