CN106554296A - A kind of Fmoc(SO with excellent hydrophilic3H) amino acid of protection and preparation method thereof - Google Patents
A kind of Fmoc(SO with excellent hydrophilic3H) amino acid of protection and preparation method thereof Download PDFInfo
- Publication number
- CN106554296A CN106554296A CN201510640945.4A CN201510640945A CN106554296A CN 106554296 A CN106554296 A CN 106554296A CN 201510640945 A CN201510640945 A CN 201510640945A CN 106554296 A CN106554296 A CN 106554296A
- Authority
- CN
- China
- Prior art keywords
- amino acid
- fmoc
- acid
- preparation
- reaction
- 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.)
- Pending
Links
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Abstract
The present invention relates to a seed amino acid protecting group, be specifically related to a kind ofly there is excellent hydrophilic Fmoc (SO for the preparation of purifying3H) protected amino acid and preparation method thereof. its primary structure is as shown in Formulae II, wherein, R is alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), proline (Pro), phenylalanine (Phe), tryptophan (Trp), methionine (Met), glycine (Gly), serine (Ser), threonine (Thr), cysteine (Cys), tyrosine (Tyr), asparagine (Asn) glutamine (Gln), lysine (Lys), arginine (Arg), histidine (His), the side chain of aspartic acid (Asp) or glutamic acid (Glu).
Description
Technical field
The present invention relates to a seed amino acid protecting group, be specifically related to a kind ofly there is excellent hydrophilic Fmo for the preparation of purifying
c(SO3H) protected amino acid and preparation method thereof.
Background technology
In recent years, adopted Fmoc to protect tactful solid-phase peptide synthetic technology application very extensive. But, owing to closing at peptideExistence in one-tenth not exclusively connects or chain termination situation, causes lacking the formation of peptide and unwanted small peptide impurity. Therefore,In order to improve the purity of polypeptide, the purifying of polypeptide just seems and is even more important. People have also improved many by the whole bag of tricksThe purity of peptide.
But, to prepare in the process of purified polypeptide at high efficiency liquid phase, some polypeptide, because its hydrophily is poor, give preparation pureChange and bring very large difficulty, this also becomes the problem that often will face in peptide purification. Therefore, the present invention is by closing at peptideIn one-tenth process, insert and there is excellent hydrophilic Fmoc (SO to its end3H) protected amino acid, thereby the parent who improves polypeptideWater-based, increases the water-soluble of polypeptide crude product, prepares purifying thereby be conducive to high efficiency liquid phase. So this have a good parentWater-based Fmoc (SO3H) introducing of protected amino acid (as reaction equation 1), for the high efficiency liquid phase preparation of polypeptide provides oneGeneral method.
InsteadAnswer formula 1 Fmoc (SO3H) end protection polypeptide is synthetic
As everyone knows, Fmoc protected amino acid is one of main selection of a-amino acid protecting group, and Fmoc protecting groupIn polypeptide is synthetic, be widely used. In the prior art, patent WO1997NL00235 has introduced Fmoc protection ammoniaBase acid and preparation method thereof. The Fmoc protected amino acid structure of its description is as Formula I:
The amino acid of Formula I: Fmoc protection
And in polypeptide is synthetic, this have an excellent hydrophilic Fmoc (SO3H) protected amino acid will have general applicationBe worth.
Thick some reason of peptide indissoluble is that Fmoc does not remove totally and causes, and by the inventive method, Fmoc is protected to ammoniaThe water miscible improvement of base acid, can make Fmoc remove incomplete probability and reduce, and increases the water-soluble of thick peptide; AnotherAspect, in thick peptide, increase residual the water miscible of Fmoc impurity, has also helped the solubility of thick peptide to increase.
Summary of the invention
Object of the present invention is in order effectively to have increased the hydrophily of polypeptide, thereby it is synthetic middle amino effectively to solve liquid phaseThe poor problem of polypeptide hydrophily in acid indissoluble and purge process.
One aspect of the present invention provides a kind of Fmoc (SO3H) amino acid of protection, its primary structure is as chemical formula
II
Formulae II: Fmoc (SO3H) amino acid of protection
Wherein, R is alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), proline (PRo), phenylalanine (Phe), tryptophan (Trp), methionine (Met), glycine (Gly), serine (Ser),Threonine (Thr), cysteine (Cys), tyrosine (Tyr), asparagine (Asn) glutamine (Gln), badThe side chain of propylhomoserin (Lys), arginine (Arg), histidine (His), aspartic acid (Asp) or glutamic acid (Glu),Preferred above-mentioned amino acid be L ?amino acid or D ?amino acid.
Another aspect of the present invention provides Fmoc (SO described above3H) the amino acid whose preparation process of protection, asLower reaction equation 2:
Reaction equation 2
Method of the present invention realizes by following steps:
Step 1: fluorenes methoxy dicarbonyl chloride (Fmoc-Cl) carries out sulfonated 9-((chloroformyl oxygen base) the methyl)-9H-fluorenes-2-sulphur that obtainsAcid;
Optionally, step 2: amino acid whose α carboxyl blocking group is protected with blocking group;
Step 3: then the amino acid of the 9-of step 1 ((chloroformyl oxygen base) methyl)-9H-fluorenes-2-sulfonic acid and α carboxy protective is enteredRow condensation reaction;
Step 4: finally carry out debenzylation reaction and can obtain Fmoc (SO3H) protected amino acid, this amino acid can with itsFmoc (SO is prepared in his amino acid or the further coupling of polypeptide fragment3H) end protection dipeptides or polypeptide.
Wherein, the reaction dissolvent of step 1 is selected from the atent solvents such as carrene, acetone, DMF or itsMixed solvent, preferably carrene; The reaction temperature of step 1 is-25 DEG C to 25 DEG C, and preferably-10 DEG C to 10 DEG C, more excellentElect as-9 DEG C ,-8 DEG C ,-7 DEG C ,-6 DEG C ,-5 DEG C ,-4 DEG C ,-3 DEG C ,-2 DEG C ,-1 DEG C, 0,1 DEG C, 2 DEG C, 3 DEG C, 4 DEG C,5 DEG C, 6 DEG C, 7 DEG C, 8 DEG C, 9 DEG C, 10 DEG C; The reaction time of step 1 is 0.5 hour to 8 hours, preferably 2 to 4Hour, more preferably 2,2.5,3,3.5,4 hours.
Wherein, the blocking group of the carboxyl in step 2 is selected from benzyl, methyl, ethyl, and it is passable that intermediate is prepared in this reactionDirectly buy, can not protect.
Wherein, the solvent in step 3 is selected from atent solvent or its mixed solvents such as ethyl acetate, carrene, DMF,Reaction temperature in step 3 is-25 DEG C to 25 DEG C, preferably-10 DEG C to 10 DEG C, and more preferably-9 DEG C ,-8 DEG C ,-7 DEG C ,-6
℃、-5℃、-4℃、-3℃、-2℃、-1℃、0、1℃、2℃、3℃、4℃、5℃、6℃、7℃、8℃、9
℃。
Wherein, the hydrogen source in step 4 is selected from ammonium formate, hydrobromic acid, formic acid etc.
Another aspect of the present invention provides a kind of solid-phase peptide synthesis, and described method comprises: by Fmoc (SO3H)After protected amino acid and coupling agent dissolve and activate in solvent, join together in solid phase reaction post, until use detection sideTill method detects reaction terminating; Wherein Fmoc (SO3H) ?A ?the A of OH represent amino acid or polypeptide, be preferably alanine(Ala), valine (Val), leucine (Leu), isoleucine (Ile), proline (Pro), phenylalanine (Phe),Tryptophan (Trp), methionine (Met), glycine (Gly), serine (Ser), threonine (Thr), cysteine(Cys), tyrosine (Tyr) asparagine (Asn), glutamine (Gln), lysine (Lys), arginine (Arg),Histidine (His), aspartic acid (Asp), glutamic acid (Glu) or comprise 1 ?100 amino acid whose peptide chains (be preferably1 ?20, more preferably 1 ?10), more preferably comprise 1 (or 2,3,4,5,6,7Individual, 8,9,10) the amino acid whose polypeptide of above slightly solubility.
Further, solid phase carrier can be the resins such as amino resins, wang resin or 2 ?Cl resins, and resin substitution degree is0.1 ?1.0mmol/g, preferably 0.2 ?0.8mmol/g, more preferably 0.2 ?0.5mmol/g.
Coupling agent of the present invention is the composition of DIC and compd A or DIPEA and compd A and compd BComposition, wherein compd A is HOBt or HOAt, compd B is PyBOP, PyAOP, HATU, HBTUOr TBTU, be preferably the composition of DIPCDI and compd A.
Further, in coupling agent the ratio of each composition taking molar ratio computing as DIPCDI:A=1.2:1.1,
DIPEA:A:B＝2.0:1.1:1.0。
In addition, the time that every seed amino acid carries out coupling reaction is generally 1.5-4 hour, preferably 2-3 hour; Pressure is preferredFor normal pressure, also can under the pressure that suitably improves or reduce, carry out; Temperature is preferably room temperature (20 ± 5 DEG C), also can beAt the temperature suitably improving or reduce, carry out.
Detailed description of the invention
The preparation of embodiment 1:9-((chloroformyl oxygen base) methyl)-9H-fluorenes-2-sulfonic acid
Fluorenes methoxy dicarbonyl chloride (64.75g) is dissolved in carrene (500ml), is cooled to 0 DEG C. Slowly drip firstThe dichloromethane solution (14mL mesyl chloride is dissolved in 250mL carrene) of sulfonic acid chloride, controls temperature and is no more than10 DEG C, drip rear continuation and stir 3 hours. In reactant liquor, add cyclohexane (500mL). Reactant liquor centrifugal treating,Gained is mixed solvent (carrene: cyclohexane=1:1,750mL × 3) point three washings for solid, obtain off-white color solid.
Structured data:
H1NMR：(D3COD)64.2-4.5(m,3,CHCH2),5.4(s,H2O/RS03H),7.1-7.9(m,6,aryl),8.1(s,
1,H-1aryl)
The preparation of embodiment 2:L-alanine benzyl ester
ALANINE (3.5g) and p-methyl benzenesulfonic acid (8.2g) are dissolved in phenmethylol (16ml), add benzene (8ml)To reactant liquor, system is warming up to 100 DEG C, benzene refluxes and is with the about 15h of water. Reactant liquor is cooled to after room temperature, adds secondEther (15ml), is then placed in refrigerator freezing and spends the night. Filter, obtain white solid. White solid acetone crystallization. ?To crystal dissolve with carrene, 5% sodium bicarbonate aqueous solution (20mL × 3) washing three times, anhydrous sodium sulfate dryingOrganic phase, filters, and revolves desolventizing, obtains colorless oil.
Embodiment 3:N-(9-(2-sulfonic acid)-fluorenylmethyloxycarbonyl)-ALANINE benzyl ester
Product 9-((chloroformyl oxygen base) the methyl)-9H-fluorenes-2-sulfonic acid (7.6g) obtaining in embodiment 1 is dissolved in ethyl acetate (10ML) and in carrene (150mL) mixed solvent. Take in addition ALANINE benzyl ester (4g) and be dissolved in carrene(100mL) and in pyridine (50ml) mixed solvent, ice bath is cooled to 0 DEG C. By 9-((chloroformyl oxygen base) methyl)-9H-Fluorenes-2-sulfonic acid solutions slowly drips in ALANINE benzyl ester solution, and temperature control is no more than 10 DEG C. After dripping, room temperature reactionDetect without raw material residual (ninhydrin colour developing) to TLC. To the adding citric acid aqueous solution (10%) washing in reactant liquor three times,Organic phase is spin-dried for, and carrene dissolves residue, washes three times. Anhydrous sodium sulfate drying organic phase, filters, is spin-dried for,Obtain yellow oil.
Embodiment 4:N-(9-(2-sulfonic acid)-fluorenylmethyloxycarbonyl)-ALANINE (Fmoc (SO3H)-L-Ala) preparation
N-(9-(2-sulfonic acid)-fluorenylmethyloxycarbonyl)-ALANINE benzyl ester (4.5g) is dissolved in methyl alcohol (30ml), then adds0.5ml glacial acetic acid, Pd/C 0.5g. Pass under hydrogen room temperature and stir 3h. Reacting liquid filtering, revolves organic solvent and obtains product.
The preparation of embodiment 5:L-phenylalanine benzyl ester
L-Phe (6.5g) and p-methyl benzenesulfonic acid (8.2g) are dissolved in phenmethylol (30ml), add benzene (15ml),System is warming up to 100 DEG C, and benzene refluxes and is with the about 15h of water. Reaction is cooled to room temperature, adds ether (30ml) to put into iceCase freeze overnight. The solid that filtration obtains acetone crystallization. The crystal obtaining dissolves with carrene, 5% sodium bicarbonate waterSolution (30mL × 3) washing, anhydrous sodium sulfate drying organic phase, filters. Revolve desolventizing, obtain yellow oil.
Embodiment 6:N-(9-(2-sulfonic acid)-fluorenylmethyloxycarbonyl)-L-Phe benzyl ester
The preparation (7.6g) of product 9-((chloroformyl oxygen base) the methyl)-9H-fluorenes-2-sulfonic acid obtaining in embodiment 1 is dissolved in acetic acidIn ethyl ester (10mL) and carrene (150mL), take in addition L-Phe benzyl ester (5.3g) and be dissolved in dichloromethaneIn alkane (100mL) and pyridine (50ml). Ice bath is cooling lower to 9-((chloroformyl oxygen base) methyl)-9H-fluorenes-2-sulfonic acid solutionsSlowly drip in L-Phe benzyl ester solution, room temperature reaction detects without raw material residual (ninhydrin colour developing) to TLC.React completely, aqueous citric acid solution (10%) washing three times, organic phase is spin-dried for, and carrene dissolves residue, washingThree times. Anhydrous sodium sulfate drying organic phase, filters, and is spin-dried for and obtains yellow oil.
Embodiment 7:N-(9-(2-sulfonic acid)-fluorenylmethyloxycarbonyl)-L-Phe (Fmoc (SO3H)-L-Phe) preparation
N-(9-(2-sulfonic acid)-fluorenylmethyloxycarbonyl)-L-Phe benzyl ester (4.5g) is dissolved in methyl alcohol (30ml), then addsEnter 0.5ml glacial acetic acid, Pd/C 0.5g. Pass under hydrogen room temperature and stir 3h. Reacting liquid filtering, revolving organic solvent must produceProduct.
The preparation of embodiment 8:L-tryptophan benzyl ester
L-Trp (8.0g) and p-methyl benzenesulfonic acid (8.2g) are dissolved in phenmethylol (30ml), add benzene (15ml)To reactant liquor, system is warming up to 100 DEG C, benzene refluxes and is with the about 15h of water. Reactant liquor is cooled to after room temperature, adds secondEther (30ml), is then placed in refrigerator freezing and spends the night. Filter, obtain white solid. White solid acetone crystallization. ?To crystal dissolve with carrene, 5% sodium bicarbonate aqueous solution (30mL × 3) washing three times, anhydrous sodium sulfate dryingOrganic phase, filters, and revolves desolventizing, obtains colorless oil.
Embodiment 9:N-(9-(2-sulfonic acid)-fluorenylmethyloxycarbonyl)-L-Trp benzyl ester
Product 9-((chloroformyl oxygen base) the methyl)-9H-fluorenes-2-sulfonic acid (7.6g) obtaining in embodiment 1 is dissolved in ethyl acetate (10ML) and in carrene (150mL) mixed solvent. Take in addition L-Trp benzyl ester (6.2g) and be dissolved in carrene(100mL) and in pyridine (50ml) mixed solvent, ice bath is cooled to 0 DEG C. By 9-((chloroformyl oxygen base) methyl)-9H-Fluorenes-2-sulfonic acid solutions slowly drips in ALANINE benzyl ester solution, and temperature control is no more than 10 DEG C. After dripping, room temperature reactionDetect without raw material residual (ninhydrin colour developing) to TLC. To the adding citric acid aqueous solution (10%) washing in reactant liquor three times,Organic phase is spin-dried for, and carrene dissolves residue, washes three times. Anhydrous sodium sulfate drying organic phase, filters, is spin-dried for,Obtain yellow oil.
Embodiment 10:N-(9-(2-sulfonic acid)-fluorenylmethyloxycarbonyl)-L-Trp (Fmoc (SO3H)-L-Trp) preparation
N-(9-(2-sulfonic acid)-fluorenylmethyloxycarbonyl)-L-Trp benzyl ester (4.5g) is dissolved in methyl alcohol (30ml), then addsEnter 0.5ml glacial acetic acid, Pd/C 0.5g. Pass under hydrogen room temperature and stir 3h. Reacting liquid filtering, revolving organic solvent must produceProduct.
N end Fmoc (SO
3
H) preparation of protection polypeptide
Fmoc-Phe-Ala-Trp-Val-OH, this polypeptide contains more hydrophobic amino acid, water-soluble poor, and purifying is comparativelyDifficulty. Therefore synthetic Fmoc (SO3H)-Phe-Ala-Trp-Val-OH, has solved polypeptide hydrophily problem, purifying more for convenience,Purity is higher. This example is embodiment, but described in patent, method is not limited only to this polypeptide.
Embodiment 11:Fmoc (SO3The preparation of H)-Phe-Ala-Trp-Val-OH polypeptide resin
Take substitution degree and be 0.310mmol/g 2 ?chlorine resin 20g, join in solid phase reaction post, with DMF washing 2Inferior, use the swelling resin of DMF after 30 minutes. Take 2.04g (6mmol) Fmoc ?Val ?OH and 0.89g (6.6mmol) DIPEAWith DMF dissolving, mixed liquor is joined in reaction column to room temperature reaction 2 hours. Reaction finishes, with DMF washing treeFat 3 times, adds DBLK deprotection 6min+8min, uses DMF washing resin 6 times, and ninhydrin detects resin color.Take 3.16g (6mmol) Fmoc ?Trp ?OH and 0.89g (6.6mmol) HOBT DMF dissolve, under ice-water bath, add 1.2After mL (7.2mmol) DIC activation 3min, mixed liquor is joined in reaction column, room temperature reaction 2 hours, with ninhydrinDetection reaction terminal is (as cessation reaction of resin water white transparency; As resin colour developing extends reaction 1 hour). Reaction finishes,With DMF washing resin 3 times, add DBLK deprotection 6min+8min, with DMF washing resin 6 times, ninhydrin inspectionAssize fat has color. Take 1.87g (6mmol) Fmoc ?Ala ?OH and 0.89g (6.6mmol) HOBT DMF dissolve, iceUnder water-bath, add after 1.2mL (7.2mmol) DIPCDI activation 3min, mixed liquor is joined in reaction column to room temperature reaction2 hours, with ninhydrin detection reaction terminal (as cessation reaction of resin water white transparency; As resin colour developing extends reaction 1Hour). Reaction finishes, and with DMF washing resin 3 times, adds DBLK deprotection 6min+8min, washs with DMFResin 6 times, ninhydrin detects resin color. Take 2.83g (6mmol) Fmoc ?Ser (tBu) ?OH and0.89g (6.6mmol) HOBT dissolves with DMF, adds after 1.2mL (7.2mmol) DIC activation 3min under ice-water bath, willMixed liquor joins in reaction column, room temperature reaction 2 hours, with ninhydrin detection reaction terminal (as resin water white transparencyCessation reaction; As resin colour developing extends reaction 1 hour). Reaction finishes, and uses DMF washing resin 3 times, adds DBLKDeprotection 6min+8min, uses DMF washing resin 6 times, and ninhydrin detects resin color. Take 2.80g (6mmol)
Fmoc(SO3H) ?Phe ?OH and 0.89g (6.6mmol) HOBT dissolve with DMF, under ice-water bath, add 1.2After mL (7.2mmol) DIC activation 3min, mixed liquor is joined in reaction column, room temperature reaction 2 hours, examines with ninhydrinMeasured reaction terminal is (as cessation reaction of resin water white transparency; As resin colour developing extends reaction 1 hour). Reaction finishes,With DMF washing resin 3 times, add DBLK deprotection 6min+8min, with DMF washing resin 6 times, ninhydrin inspectionAssize fat has color. Coupling finishes, and resin shrinkage is drained, and obtains 22.68 grams of peptide resins.
Embodiment 12:Fmoc (SO3The preparation of H)-thick peptide of Phe-Ala-Trp-Val-OH
22.68 grams of peptide resins that embodiment 11 is obtained join in 500ml single port bottle, add in advance and prepareTFA:H2O:EDT:PhOMe: thioanisole=90:5:2:2:1 (V:V) 280ml, room temperature reaction 2 hours,Filter resin, collect filtrate. With a small amount of TFA washing resin, merging filtrate. Filtrate is slowly added to 2800ml ice secondIn ether, precipitate, centrifugal, ice ether washing 5 times, drying under reduced pressure obtains 4.58 grams of thick peptides, HPLC purity 81.6%.
Embodiment 13:Fmoc (SO3The preparation of H)-Phe-Ala-Trp-Val-OH essence peptide
4.58 grams of thick peptides that embodiment 12 is obtained are used high efficiency liquid phase purification. Taking anti-phase octadecylsilane as fixing phase,Taking 0.1% aqueous acetic acid/acetonitrile as mobile phase, collect object peak cut, concentrated freeze-dried, obtain sterling 3.0g, purity is largeIn 99%.
Abbreviation in the present invention and English implication
Claims (10)
1. a Fmoc (SO3H) amino acid (Fmoc (SO of protection3H)-A-OH), its primary structure asFormulae II
Wherein, R is alanine (Ala), valine (Val), leucine (Leu), isoleucine (Ile), dried meatPropylhomoserin (Pro), phenylalanine (Phe), tryptophan (Trp), methionine (Met), glycine (Gly), silkPropylhomoserin (Ser), threonine (Thr), cysteine (Cys), tyrosine (Tyr), asparagine (Asn) paddyGlutamine (Gln), lysine (Lys), arginine (Arg), histidine (His), aspartic acid (Asp)Or the side chain of glutamic acid (Glu).
2. a Fmoc (SO according to claim 13H) the amino acid whose preparation method of protection, itsComprise the steps:
Step 1: fluorenes methoxy dicarbonyl chloride (Fmoc-Cl) carries out sulfonated 9-((chloroformyl oxygen base) the methyl)-9H-that obtainsFluorenes-2-sulfonic acid;
Optionally, step 2: amino acid whose α carboxyl blocking group is protected with blocking group;
Step 3: then by the ammonia of the 9-of step 1 ((chloroformyl oxygen base) methyl)-9H-fluorenes-2-sulfonic acid and α carboxy protectiveCondensation reaction is carried out in base acid;
Step 4: the blocking group on amino acid whose α carboxyl is removed and obtain Fmoc (SO3H) protection ammoniaBase acid.
3. preparation method according to claim 2, wherein, the reaction dissolvent of step 1 be selected from carrene,The atent solvent such as acetone, DMF or its mixed solvent, preferably carrene; Step 1 anti-Answering temperature is-25 DEG C to 25 DEG C, preferably-10 DEG C to 10 DEG C, and more preferably-9 DEG C ,-8 DEG C ,-7 DEG C ,-6 DEG C ,-5
℃、-4℃、-3℃、-2℃、-1℃、0、1℃、2℃、3℃、4℃、5℃、6℃、7℃、8℃、9DEG C, 10 DEG C; The reaction time of step 1 is 0.5 hour to 8 hours, preferably 2 to 4 hours, more preferably 2,2.5,3,3.5,4 hours.
4. according to the preparation method described in claim 2-3 any one, wherein, the protection of the carboxyl in step 2Group is selected from benzyl, methyl, ethyl.
5. according to the preparation method described in claim 2-4 any one, wherein, the solvent in step 3 is selected from secondAtent solvent or its mixed solvents such as acetoacetic ester, carrene, DMF, the reaction temperature in step 3 is-25DEG C to 25 DEG C, preferably-10 DEG C to 10 DEG C, more preferably-9 DEG C ,-8 DEG C ,-7 DEG C ,-6 DEG C ,-5 DEG C ,-4 DEG C ,-3
℃、-2℃、-1℃、0、1℃、2℃、3℃、4℃、5℃、6℃、7℃、8℃、9℃。
6. according to the preparation method described in claim 2-5 any one, wherein, the hydrogen source in step 4 is selected from firstAcid ammonium, hydrobromic acid, formic acid etc.
7. a solid-phase peptide synthesis, described method comprises: by Fmoc (SO3H) protected amino acid
(Fmoc(SO3H) ?A ?OH) and after coupling agent dissolves and activate in solvent, join together in solid phase reaction post,Until reaction terminating detected by detection method;
Wherein, Fmoc (SO3H) ?A ?the A of OH represent amino acid or peptide chain, preferably represent alanine (Ala),Valine (Val), leucine (Leu), isoleucine (Ile), proline (Pro), phenylalanine (Phe),Tryptophan (Trp), methionine (Met), glycine (Gly), serine (Ser), threonine (Thr), halfCystine (Cys), tyrosine (Tyr) asparagine (Asn), glutamine (Gln), lysine (Lys),Arginine (Arg), histidine (His), aspartic acid (Asp), glutamic acid (Glu) or comprise 1 ?20 (1 ?10)Individual amino acid whose peptide chain, more preferably comprises the amino acid whose polypeptide of more than 1 slightly solubility.
8. synthetic method according to claim 7, wherein, the filler in solid phase reaction post be amino resins,Wang resin or 2 ?the resin such as Cl resin, resin substitution degree be 0.1 ?1.0mmol/g, preferably 0.2 ?0.8mmol/g,More preferably 0.2 ?0.5mmol/g.
9. coupling agent of the present invention is composition or DIPEA and compd A and the change of DIC and compd AThe composition of compound B, wherein compd A is HOBt or HOAt, compd B be PyBOP, PyAOP,HATU, HBTU or TBTU, be preferably the composition of DIPCDI and compd A, more electedly, and couplingIn agent, the ratio of each composition is taking molar ratio computing as DIPCDI:A=1.2:1.1, DIPEA:A:B=2.0:1.1:1.0.
10. an amino protecting agent for hydrophilic amino acid or polypeptide, the ammonia of described hydrophilic amino acid or polypeptideBase protective agent is 9-((chloroformyl oxygen base) methyl)-9H-fluorenes-2-sulfonic acid, preferably, and 9-((chloroformyl oxygen base) methyl)-9H-fluorenes-2-sulfonic acid is for increasing amino acid or the hydrophilic purposes of polypeptide.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510640945.4A CN106554296A (en) | 2015-09-30 | 2015-09-30 | A kind of Fmoc(SO with excellent hydrophilic3H) amino acid of protection and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510640945.4A CN106554296A (en) | 2015-09-30 | 2015-09-30 | A kind of Fmoc(SO with excellent hydrophilic3H) amino acid of protection and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106554296A true CN106554296A (en) | 2017-04-05 |
Family
ID=58417892
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510640945.4A Pending CN106554296A (en) | 2015-09-30 | 2015-09-30 | A kind of Fmoc(SO with excellent hydrophilic3H) amino acid of protection and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106554296A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111499687A (en) * | 2020-03-13 | 2020-08-07 | 宁波市第一医院 | Sphingosine-1-phosphate receptor 3 agonist, and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4293455A (en) * | 1980-04-07 | 1981-10-06 | Rockefeller University | N.sup.α -Desacetylthymosinα1 and process |
| WO2002036067A2 (en) * | 2000-11-01 | 2002-05-10 | Yeda Research And Development Co. Ltd. | Long-acting cytokine derivatives and pharmaceutical compositions comprising them |
| WO2004089279A2 (en) * | 2003-04-08 | 2004-10-21 | Yeda Research And Development Co. Ltd. | Long-acting derivatives of pyy agonists |
| CN102558338A (en) * | 2012-02-10 | 2012-07-11 | 深圳翰宇药业股份有限公司 | Method for synthesizing lixisenatide |
-
2015
- 2015-09-30 CN CN201510640945.4A patent/CN106554296A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4293455A (en) * | 1980-04-07 | 1981-10-06 | Rockefeller University | N.sup.α -Desacetylthymosinα1 and process |
| WO2002036067A2 (en) * | 2000-11-01 | 2002-05-10 | Yeda Research And Development Co. Ltd. | Long-acting cytokine derivatives and pharmaceutical compositions comprising them |
| WO2004089279A2 (en) * | 2003-04-08 | 2004-10-21 | Yeda Research And Development Co. Ltd. | Long-acting derivatives of pyy agonists |
| CN102558338A (en) * | 2012-02-10 | 2012-07-11 | 深圳翰宇药业股份有限公司 | Method for synthesizing lixisenatide |
Non-Patent Citations (1)
| Title |
|---|
| R. B. MERRIFIELD ET AL.: "9-(2-Sulfo)fluorenylmethyloxycarbonyl Chloride, a New Reagent for the Purification of Synthetic Peptides", 《JOURNAL OF THE ORGANIC CHEMISTRY》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111499687A (en) * | 2020-03-13 | 2020-08-07 | 宁波市第一医院 | Sphingosine-1-phosphate receptor 3 agonist, and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103980358B (en) | A kind of method preparing Arg34Lys26-(N-EPSILON-(N-ALPHA-Palmitoyl-L-GAMMA-glutamyl))-GLP-1[7-37] | |
| EP3398960B1 (en) | Method for preparing semaglutide | |
| US3853837A (en) | Novel nonapeptide amide analogs of luteinizing hormone releasing factor | |
| WO2018033127A1 (en) | Synthesis method for low-racemization impurity liraglutide | |
| DE CASTIGLIONE et al. | Synthesis of dermorphins, a new class of opiate‐like peptides | |
| WO2017020569A1 (en) | Synthesis-separation purification method for indissolvable polypeptide | |
| CN104387454A (en) | Method for preparing triptorelin by using fragment condensation | |
| Pallai et al. | Partial retro-inverso analogs of somatostatin: pairwise modifications at residues 7 and 8 and at residues 8 and 9 | |
| CN102875648B (en) | Method for preparing telaprevir | |
| CN106554391B (en) | Method for synthesizing marine biological peptide Xen2174 | |
| CN104177490A (en) | Method for preparing salmon calcitonin acetate by fragment condensation | |
| Katakai | Peptide synthesis using o-nitrophenylsulfenyl N-carboxy. alpha.-amino acid anhydrides | |
| CN106554296A (en) | A kind of Fmoc(SO with excellent hydrophilic3H) amino acid of protection and preparation method thereof | |
| CN104211801A (en) | Method for preparing lixisenatide | |
| JPS5973574A (en) | Cyclic dipeptide | |
| EP2864347B1 (en) | Synthesis of beta-turn peptidomimetic cyclic compounds | |
| EP3914605A1 (en) | Peptide precipitation method | |
| Manne et al. | Incorporation of pseudoproline monomer (Fmoc-Thr [ψMe, Mepro]–OH) facilitates efficient solid-phase synthesis of difficult peptides | |
| EP0124420A2 (en) | Gastric secretion inhibiting peptide derivatives, process for their preparation and medicines containing them | |
| Ariyoshi et al. | The synthesis of a sweet peptide, α-l-aspartyl-l-phenylalanine methyl ester, without the use of protecting groups | |
| WO1992001709A1 (en) | Process for synthesizing peptides | |
| Katsoyannis et al. | Synthesis of destetrapeptide B27-30 human (porcine) insulin. Biologically active insulin analog | |
| JPH08119916A (en) | Selective production of n-protected glutamic acid gamma-derivative | |
| CN106317161B (en) | Preparation method of fluoromethyl ketone peptide series compounds | |
| RU2592282C1 (en) | Method of producing nonapeptides |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170405 |
|
| RJ01 | Rejection of invention patent application after publication |