WO2015147138A1 - Procédé de production de peptide - Google Patents

Procédé de production de peptide Download PDF

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Publication number
WO2015147138A1
WO2015147138A1 PCT/JP2015/059317 JP2015059317W WO2015147138A1 WO 2015147138 A1 WO2015147138 A1 WO 2015147138A1 JP 2015059317 W JP2015059317 W JP 2015059317W WO 2015147138 A1 WO2015147138 A1 WO 2015147138A1
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WIPO (PCT)
Prior art keywords
peptide
amino acid
acid
ionic liquid
liquid
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PCT/JP2015/059317
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English (en)
Japanese (ja)
Inventor
真也 古川
高英 福山
日馨 柳
隆 稲山
遼太朗 中谷
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味の素株式会社
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Priority to JP2016510472A priority Critical patent/JP6579390B2/ja
Publication of WO2015147138A1 publication Critical patent/WO2015147138A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/02—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length in solution

Definitions

  • the present invention relates to a method for producing a peptide for separating a peptide from a liquid mixture of ionic liquid containing peptide and amino acid, and in particular, a method for producing a peptide for efficiently isolating a peptide synthesized using an ionic liquid from a liquid mixture, In particular, it relates to a method for industrial production.
  • Patent Document 1 discloses a method in which an ionic liquid and an oligopeptide, oligosaccharide, or oligonucleotide are bound to improve solubility in an organic solvent and also used as a protective group. In the polymerization of each reaction, a protecting group and a condensing agent are used.
  • Non-Patent Document 1 discloses Z-Asp (Z-Aspartic acid) + PM (L-Phenylalanine methylester) ⁇ Z-APM (Z-) in an ionic liquid (BP6 (1-butyl-3-methylimidazolium hexafluorophosphate)). Aspartame) is performed using an enzyme (Thermolysin), and it has been demonstrated that the enzyme reaction can be performed even in an ionic liquid. The yield in this reaction is as high as 90%, but it is extremely low, and the reaction method disclosed here is merely a solvent substitution of the enzyme reaction in an organic solvent.
  • Patent Document 2 discloses peptide synthesis in an ionic liquid (4-methyl-N-butylpyridinium tetrafluoroborate).
  • the synthesis method described here is a simple solvent replacement of an enzyme reaction in an organic solvent as in Non-Patent Document 1, and is a reaction at a considerably low concentration of 20 mM, and also uses a protecting group.
  • Non-Patent Document 2 demonstrates that amino acids can be bound to ionic liquid residues by ionic bonds, and that amino acids can be made ionic liquid. In particular, its use is not discussed. Are considering.
  • Non-Patent Document 3 describes a review of polypeptides, oligosaccharides, and other organic synthesis using ionic liquids, and there is a description that the substrate-ionic liquid is used as a reaction intermediate in the gist. No specific data is shown.
  • Non-Patent Document 4 describes reviews such as introductions regarding amino acid-binding ionic liquids, and there is no specific data, but mentions future use as a solvent or catalyst.
  • the peptide synthesis method using the proposed ionic liquid has a low yield of the obtained peptide, and is not sufficient as an industrial peptide production method. Under such circumstances, a production method for synthesizing a peptide with high yield using an ionic liquid has been proposed (Patent Documents 3 and 4).
  • JP 2008-537733 A JP 2008-301829 A WO2012 / 014808 publication WO2012 / 014809
  • the present invention relates to a liquid mixture comprising a peptide and an amino acid, wherein the peptide is separated from the liquid mixture in which at least one selected from the peptide and amino acid is present as an ionic liquid in which a cation and an ionic bond are formed as anions.
  • an object of the present invention is to provide a peptide production method for efficiently isolating a peptide synthesized using an ionic liquid from a liquid mixture.
  • the present invention when a liquid mixture containing a peptide synthesized using an ionic liquid is treated with an acid, preferably in an organic solvent, an unreacted amino acid and a peptide as a product can be precipitated and recovered stepwise. It was made based on the knowledge that it can be done. That is, the present invention includes the following contents.
  • a method for producing a peptide characterized by preferentially precipitating or separating an amino acid, or preferentially precipitating and separating an amino acid, followed by further acid treatment to preferentially precipitate and separate the peptide.
  • the method according to [1] wherein the liquid mixture is acid-treated, first the amino acids are preferentially precipitated and separated, and then further acid-treated to preferentially precipitate and separate the peptides.
  • the liquid mixture is a liquid mixture containing (1) an ionic liquid (ionic liquid A) having a peptide as an anion and (2) an ionic liquid (ionic liquid B) having an amino acid as an anion, [1] or [2 ]the method of.
  • the condensation reaction is a reaction in which (2) an amino group of an ionic liquid (ionic liquid B) having an amino acid as an anion and (3) an ester group of a peptide ester or amino acid ester are condensed to form an amide bond.
  • the amino acid is first precipitated by the acid treatment, and the acid treatment of the liquid mixture is performed in the presence of 50 to 100 mol% of acid with respect to the molar amount in the liquid mixture of the previously precipitated amino acid.
  • One of the methods of [7]. [10] The method according to any one of [1] to [9], wherein the liquid mixture to be acid-treated contains an organic solvent.
  • the ionic liquid is ionized by forming an ionic bond with at least one cation selected from alkylphosphonium ions, alkylimidazolium ions, alkylammonium ions, alkylpyridinium ions, alkylpyrrolidinium ions, and alkylpiperidinium ions.
  • the method according to any one of [1] to [14], which is liquefied.
  • a method for separating a peptide and an amino acid, wherein the amino acid is preferentially precipitated is also known in the art.
  • a liquid comprising a first amino acid or peptide and a second amino acid or peptide different from the first amino acid or peptide, wherein at least one of the first amino acid or peptide and the second amino acid or peptide
  • One kind is an acid treatment of the liquid present as an ionic liquid that forms an ionic bond with a cation as an anion, and preferentially precipitates and separates either the first amino acid or peptide and the second amino acid or peptide
  • a liquid comprising a first amino acid or peptide and a second amino acid or peptide different from the first amino acid or peptide, wherein at least one of the first amino acid or peptide and the second amino acid or peptide
  • One kind is an acid treatment of the liquid present as an ionic liquid that forms an ionic bond with a cation as an anion, and preferentially precipitates and separates either the first amino acid or peptide and the second amino acid or peptide
  • a method for separating a first amino acid or peptide from a second amino acid or peptide which is characterized.
  • a liquid mixture containing a peptide and an amino acid, wherein at least one selected from the peptide and amino acid forms an ionic bond with a cation as an anion to form an ionic liquid a simple mixture is obtained.
  • the peptide can be selectively precipitated and separated efficiently and with high yield.
  • the product peptide can be selectively precipitated and separated. Therefore, the present invention can easily separate an amino acid and a peptide. Therefore, the peptide production method of the present invention is a particularly excellent industrial production method.
  • a liquid mixture containing a peptide and an amino acid wherein the liquid mixture containing at least one selected from the peptide and amino acid as an anion and forming an ionic bond with a cation is acid-treated, After preferentially precipitating or separating the amino acid, preferentially precipitating and separating the amino acid, further acid treatment to preferentially precipitate and separate the peptide.
  • the liquid mixture of the present invention includes peptides and amino acids. At least one of the peptides and amino acids contained in the liquid mixture forms an ionic liquid. In this case, the amino acid or peptide that forms the ionic liquid becomes an anion, and is formed into an ionic liquid by forming an ionic bond with the cation. Both amino acids and peptides may be ionic liquids.
  • the ionic liquidized peptide and / or ionic liquidized amino acid contained in the liquid mixture is precipitated and separated as a peptide and / or amino acid in a form in which the ionic liquidization is released by acid treatment.
  • the amino acid is first preferentially precipitated by acid treatment of the liquid mixture.
  • the amino acid is precipitated and separated, it is further acid-treated to precipitate and separate the peptide preferentially.
  • the peptide may be isolated as it is.
  • preferentially precipitating and separating amino acids means that at least the ratio of the solid amino acid to peptide molar ratio or mass ratio (preferably the molar ratio) obtained by precipitation separation is higher for amino acids.
  • the molar ratio of amino acid: peptide is preferably 2: 1 or more, more preferably 5: 1 or more, more preferably 10: 1 or more, more preferably 15: 1 or more, more preferably 20: 1 or more.
  • preferentially precipitating and separating a peptide means that the peptide has a larger ratio of at least a molar ratio or a mass ratio (preferably a molar ratio) between a solid amino acid and a peptide obtained by precipitation separation.
  • the molar ratio of amino acid: peptide is 1: 1.2 or more, more preferably 1: 1.5 or more, more preferably 1: 2 or more, more preferably 1: 2.5 or more, more preferably 1: 3. That's it.
  • the liquid mixture includes (1) an ionic liquid having a peptide as an anion (ionic liquid A) and (2) an ionic liquid having an amino acid as an anion (ionic liquid B).
  • the liquid mixture is a reaction solution in which a peptide is produced by a condensation reaction.
  • condensation reaction and reaction solution typically include the condensation reaction described in WO2012 / 014808 and WO2012 / 014809 and the reaction solution obtained thereby. Descriptions of these patent publications are included in the description of this specification.
  • the liquid mixture is an amide bond formed by condensation reaction of (2) an amino group of an ionic liquid (ionic liquid B) having an amino acid as an anion and (3) an ester group of a peptide ester or amino acid ester.
  • ionic liquid B an amino group of an ionic liquid having an amino acid as an anion
  • ester group of a peptide ester or amino acid ester In the case of a reaction solution in which is formed.
  • Such a condensation reaction can be carried out according to the method described in WO2012 / 014809.
  • the condensation reaction is performed using an ionic liquid (ionic liquid B) having an amino acid as a reaction raw material as an anion as a reaction solvent.
  • ionic liquid B having an amino acid as a reaction raw material as an anion as a reaction solvent.
  • the condensation is performed in the presence of 20% by mass or less of water based on the total mass of the reaction system.
  • the peptide When the peptide is first precipitated by the acid treatment of the mixed solution, it is preferably 25 to 200 mol%, more preferably 30 to 150 mol%, still more preferably 50 to the molar amount of the peptide present in the mixed solution. Acid treatment with ⁇ 100 mol% acid precipitates and separates the peptide.
  • the peptide referred to herein includes those existing in the form of a salt, such as an ionic liquid peptide.
  • the amino acid When the amino acid first precipitates due to the acid treatment of the mixed solution, it is preferably 25 to 200 mol%, more preferably 30 to 150 mol%, more preferably 50 to 50 mol% with respect to the molar amount of the amino acid present in the mixed solution.
  • the acid treatment is preferably performed with an acid of preferably 50 to 300 mol%, more preferably 50 to 200 mol%, based on the molar amount of the peptide present in the mixed solution.
  • the peptide referred to herein includes those existing in the form of a salt, such as an ionic liquid peptide.
  • amino acids herein include those that exist in the form of salts, such as amino acids that are ionic liquids.
  • the acid treatment is preferably performed with an acid of 25 to 200 mol%, more preferably 30 to 150 mol%, and still more preferably 50 to 100 mol% with respect to the molar amount in the mixed solution of the peptide first precipitated by the acid treatment.
  • the acid treatment is preferably performed with an acid of 25 to 200 mol%, more preferably 30 to 150 mol%, and still more preferably 50 to 100 mol% with respect to the molar amount in the mixed solution of the peptide first precipitated by the acid treatment.
  • it is further acid-treated to preferentially separate and separate other peptides.
  • the acid is preferably used in an acid amount of preferably 50 to 300 mol%, more preferably 50 to 200 mol%, based on the molar amount of the other peptide present in the mixed solution.
  • the peptide referred to herein includes those existing in the form of a salt, such as an ionic liquid peptide.
  • the liquid mixture preferably contains an organic solvent.
  • the viscosity is generally high and is often not suitable for precipitation separation of the target product by acid treatment. Therefore, it is preferable to lower the viscosity with an organic solvent to make a liquid mixture suitable for precipitation separation of the target product.
  • the condensation reaction is performed using an ionic liquid as a reaction solvent, and when the reaction solution does not contain an organic solvent, it is preferable to add an organic solvent and perform acid treatment.
  • acid treatment may be performed as it is without adding the organic solvent to the reaction solution (liquid mixture).
  • the organic solvent may be added again before acid treatment of the liquid mixture after the precipitation and separation of the amino acid.
  • it may be used as it is without adding an organic solvent.
  • Subsequent acid treatment may be performed after solvent replacement, such as by distilling off the organic solvent under reduced pressure and adding another organic solvent.
  • the organic solvent is not particularly limited as long as it is an organic solvent that dissolves in the liquid mixture and does not decompose the ionic liquid.
  • organic solvent examples include hydrocarbons, ketones, halogenated hydrocarbons, nitrified hydrocarbons, ethers, esters, alcohols, N, N-dimethylformamide, dimethyl sulfoxide and the like.
  • hydrocarbon examples include benzene, toluene, xylene, solvent naphtha, normal hexane, isohexane, cyclohexane, methylcyclohexane, normal heptane, isooctane, and normal decane.
  • ketone examples include acetone, methyl ethyl ketone, methyl isobutyl ketone, diisobutyl ketone, cyclohexanone, diacetone alcohol and the like.
  • the halogenated hydrocarbon is preferably one having 1 to 6 carbon atoms, wherein a part or all of the hydrogen atoms bonded to the carbon atom are substituted with a halogen element. Specific examples include chloroform, dichloromethane, trichloroethylene, tetrachloroethylene, 1,1-dichloro 1-fluoroethane, dichloropentafluoropropane, 1-bromopropane, and o-dichlorobenzene.
  • nitrified hydrocarbon a compound in which a nitrile group is bonded to a hydrocarbon having 1 to 6 carbon atoms is preferable, and examples thereof include acetonitrile.
  • ethers include diisopropyl ether, 1,4-dioxane, methyl tert-butyl ether, 1,2-dimethoxyethane, diethyl ether, and tetrahydrofuran.
  • ester examples include ethyl formate, propyl formate, methyl acetate, ethyl acetate, propyl acetate, butyl acetate, methoxybutyl acetate, amyl acetate, normal propyl acetate, isopropyl acetate, methyl lactate, ethyl lactate, and butyl lactate.
  • the alcohol examples include methanol, ethanol, butanol, isopropyl alcohol, normal propyl alcohol, 2-butanol, isobutanol, tert-butanol, butanediol, ethylhexanol, benzyl alcohol and the like.
  • Examples of other solvents include N, N-dimethylformamide and dimethyl sulfoxide.
  • An organic solvent may be used individually by 1 type, and 2 or more types may be mixed and used for it.
  • the organic solvent is preferably an aprotic solvent, and particularly preferable organic solvents include chloroform, acetonitrile, tetrahydrofuran, ethyl acetate, or any mixed solvent thereof.
  • an acid treatment is performed by addition of the compound which generate
  • the amount of the organic solvent added in the present invention is not particularly limited as long as the liquid mixture is suitable for precipitation separation of amino acids and peptides. Usually, it is preferable to add 20 to 500 parts by volume, more preferably 200 to 400 parts by volume of the organic solvent per 100 parts by mass of the liquid mixture.
  • the amount of water in the liquid mixture is preferably 20% by mass or less, more preferably 10% by mass or less, and more preferably 0 to 5% by mass with respect to the total mass of the liquid mixture. preferable.
  • examples of the acid used for the acid treatment include inorganic acids such as hydrogen chloride, sulfuric acid, nitric acid, phosphoric acid, boric acid and hydrofluoric acid, and organic acids such as formic acid, acetic acid and propionic acid. . These acids may be used individually by 1 type, and 2 or more types may be mixed and used for them.
  • the acid treatment can be performed by adding these acids to the liquid mixture. Moreover, you may carry out by adding the compound which generate
  • TMSCl chlorotrimethylsilane
  • a compound that promotes acid generation from the compound is used together with a compound that generates an acid by reaction.
  • a compound that promotes acid generation it is preferable to use a lower alcohol having 1 to 4 carbon atoms. Specific examples include methanol, ethanol, n-propanol, isopropanol, and butanol.
  • the organic solvent in the method of the present invention contains a sufficient amount for acid generation, addition as a compound for promoting acid generation can be omitted.
  • the acid treatment is preferably added to the liquid mixture in the presence of an organic solvent such as after addition of the organic solvent.
  • an organic solvent can be added to the liquid mixture when no organic solvent is present, and an acid or a compound that generates an acid by reaction can be added.
  • the liquid mixture When the amino acid or peptide is precipitated, it is preferable to cool the liquid mixture in order to increase the precipitation efficiency. Cooling is preferably carried out after adding an acid to the liquid mixture or after generating an acid with a compound that generates an acid by reaction.
  • the cooling temperature is preferably 10 ° C. or lower. It does not specifically limit as a method to isolate
  • the target peptide separated as described above can be further purified to improve the purity by a known method such as recrystallization.
  • the separated peptide (solid) is dissolved in water or the like, and a lower alcohol such as methanol is gradually added to crystallize a highly pure peptide.
  • the ionic liquid contained in the liquid mixture is made into an ionic liquid by using a peptide and / or amino acid as an anion and forming an ionic bond with the cation.
  • the amino acid that becomes an anion is not particularly limited, and may be, for example, a neutral amino acid, an acidic amino acid, or a basic amino acid. .
  • the amino acids are leucine (Leu), phenylalanine (Phe), tyrosine (Tyr), glycine (Gly), alanine (Ala), valine (Val), isoleucine (Ile), serine (Ser), lysine (Lys), Proline (Pro), Threonine (Thr), Methionine (Met), Glutamic acid (Glu), Cysteine (Cys), Aspartic acid (Asp), Glutamine (Gln), Tryptophan (Trp), Histidine (His), Arginine (Arg) , Amino acids such as hydroxyproline, hydroxylysine, N-methylglycine, ornithine and the like.
  • ⁇ -amino acids such as ⁇ -alanine
  • ⁇ -amino acids such as ⁇ -aminobutyric acid
  • unnatural amino acids in which the side chain of these amino acids is modified with an organic group
  • amino acids may be either L-form or D-form.
  • neutral amino acids specifically, amino acids, such as glycine, alanine, valine, isoleucine, and leucine, can be mentioned.
  • the peptide that becomes an anion is not particularly limited.
  • the amino acid exemplified above is used as a residue.
  • examples include oligopeptides.
  • the number of residues of the peptide is not particularly limited, but is preferably a peptide of 5 or less, more preferably 3 or less, and still more preferably 2 residues.
  • Dipeptides include glycylglycine, glycylalanine, alanylglycine, valylglycine, alanylalanine, alanylvaline, prolylglycine, valylalanine, glycylphenylalanine, alanylphenylalanine and the like.
  • the functional group present in the side chain or the like in these amino acids or peptides may be protected with a protecting group generally used in peptide chemistry or the like as long as they can form an ionic liquid as an anion.
  • the amino group in the side chain may be protected with an amino protecting group such as formyl group, benzyloxycarbonyl group or tert-butoxycarbonyl group.
  • the side chain carboxy group is methyl, ethyl, tert-butyl, benzyl, trimethylsilyl, triethylsilyl, dimethylphenylsilyl, tert-butyldimethylsilyl, tert-butyldiethylsilyl, etc.
  • the hydroxyl group of the side chain is benzyl group, acetyl group, chloroacetyl group, benzoyl group, benzylcarbonyl group, 2-tetrahydropyranyl group, 2-tetrahydrofuranyl group, trimethylsilyl group, triethylsilyl group, dimethylphenylsilyl group, It may be protected with a protecting group such as a tert-butyldimethylsilyl group or a tert-butyldiethylsilyl group.
  • the amino acid and peptide in the present invention include a form in which the functional group present in the molecule is protected with a protecting group.
  • a compound having a quaternized hetero atom for example, a quaternary phosphonium salt, a quaternary ammonium salt, an imidazolium salt, a pyridinium salt, a pyrrolidinium salt, a piperidinium salt, etc.
  • cations include a cation that is at least one selected from alkylphosphonium ions, alkylimidazolium ions, alkylammonium ions, alkylpyridinium ions, alkylpyrrolidinium ions, and alkylpiperidinium ions.
  • the alkyl group in the alkylphosphonium ion or the like used here preferably has 1 to 12 carbon atoms, more preferably 1 to 6 carbon atoms, and most preferably 1 to 4 carbon atoms.
  • a plurality of alkyl groups may be the same or different, but are preferably the same.
  • the ionic liquid is not a salt melt but a salt composed of ions that melt at a low temperature of 100 ° C. or lower. Therefore, water is not an ionic liquid.
  • the compound having the quaternized heteroatom and the first amino acid or peptide are mixed in approximately equimolar amounts and heated under non-reduced pressure or reduced pressure (preferably 20 to 150 mmHg) (preferably 40 to 70 ° C.), water is evaporated and dehydration condensation can be performed to prepare an amino acid or peptide that has been made into an ionic liquid.
  • the ionic liquidized amino acid or peptide is carboxylate, that is, the carboxy group in the first amino acid or peptide forms an ionic bond with the compound having the quaternized hetero atom. preferable.
  • Non-patent document 2 (Acc. Chem. Res. 2007, 40, 1122-1129) about compounds having quaternized heteroatoms, amino acids and peptides, ionic liquidized amino acids or peptides is described in the present specification. It shall be included in the description.
  • the liquid mixture is a reaction solution in which a peptide is produced by a condensation reaction
  • a reaction solution typically includes a reaction solution obtained by the condensation reaction described in WO2012 / 014808 and WO2012 / 014809.
  • the liquid mixture is particularly a reaction solution obtained by the condensation reaction described in WO2012 / 014809, which contains a peptide of interest and an amino acid as an impurity, and at least one selected from the peptide and amino acid is a cation as an anion And a liquid mixture which exists as an ionic liquid in which an ionic bond is formed.
  • a liquid mixture obtained by condensation reaction with an amino acid ester or peptide ester using an ionic liquid having an amino acid or peptide as an anion as a reaction solvent and a reaction raw material is preferable.
  • the liquid mixture does not include a liquid mixture obtained by a condensation reaction between an ionic liquid having a peptide as an anion and a peptide ester, that is, a liquid mixture containing no amino acid.
  • the ionic liquid having an amino acid or peptide as an anion a substance having the same meaning as the ionic liquid having an amino acid or peptide described above as an anion and forming an ionic bond with a cation can be used.
  • a peptide ester or amino acid ester ester group and an amino group are subjected to a condensation reaction to form a peptide bond, and to obtain the target peptide, an amino group bonded to the ⁇ , ⁇ or ⁇ carbon atom of the amino acid, Alternatively, it is preferable to use an amino group which is not protected at the N-terminal of the peptide.
  • the C-terminal carboxy group of the peptide described above or a peptide synonymous with an amino acid is esterified with an alkyl group or the like.
  • the alkyl group preferably has 1 to 12 carbon atoms, more preferably 1 to 6 carbon atoms, and particularly preferably 1 to 4 carbon atoms.
  • the amino group bonded to the N-terminal amino group of the peptide ester or the ⁇ , ⁇ , or ⁇ carbon atom of the amino acid ester may be protected with a protective group, but it is preferable to use an amino group that is not protected.
  • the condensation reaction can be performed by a method according to the condensation reaction described in WO2012 / 014809.
  • an ionic liquidized amino acid or peptide can be reacted with an amino acid ester or peptide ester using a reaction solvent and a reaction raw material.
  • the ionic liquidized amino acid is used in an equimolar amount or more with respect to the amino acid ester or peptide ester, and preferably the molar ratio of the ionic liquid amino acid or peptide: amino acid ester or peptide ester is 20: It is used at 1: 1 to 1: 1, more preferably 10: 1 to 2: 1.
  • a peptide bond is formed between the amino group of the amino acid or peptide that has been liquefied and the carboxy group cleaved from the amino acid ester or ester of the peptide ester.
  • Alcohol is produced from the alcohol residue constituting the ester.
  • a peptide hydrolase or a condensing agent may not be present.
  • the amount of water is preferably 20% by mass or less, more preferably 10% by mass or less, and further preferably 0 to 5% by mass with respect to the total mass of the reaction system.
  • the condensation reaction is preferably carried out by mixing both and maintaining the temperature at 0 to 100 ° C., preferably at room temperature (20 ° C.) to 70 ° C.
  • the reaction time is not particularly limited, and is usually 10 minutes to 3 days, preferably 30 minutes to 2 days, and more preferably 1 hour to 24 hours.
  • the liquid mixture includes an amino group of an ionic liquid (ionic liquid B) having an amino acid as an anion and (3) an ester group of a peptide ester or an amino acid ester.
  • ionic liquid B an amino group of an ionic liquid having an amino acid as an anion
  • ester group of a peptide ester or an amino acid ester Is a reaction solution in which an amide bond is formed by a condensation reaction.
  • liquid mixture used in the present invention include, for example, a first amino acid or peptide (A), a second amino acid or peptide (B), and a peptide hydrolase (C) that are ionic liquidized.
  • first amino acid or peptide in the form of ionic liquid is used as a reaction solvent and as a reaction raw material to form a peptide bond between the first amino acid or peptide and the second amino acid or peptide.
  • the liquid mixture obtained by making it can be mentioned.
  • the embodiment in which the component (A) and the component (B) simultaneously become a peptide in the condensation reaction is excluded.
  • the condensation reaction can be carried out by a condensation reaction with a second amino acid using an ionic liquid having an amino acid or peptide as an anion as a reaction solvent and a reaction raw material.
  • the condensation reaction can be carried out by a condensation reaction with a second amino acid or peptide using an ionic liquid having an amino acid as an anion as a reaction solvent and a reaction raw material. Groups that do not participate in the reaction may be protected with a protecting group.
  • Such a condensation reaction can be carried out according to the method described in WO2012 / 014808. In the condensation reaction, it is preferable that 50% by mass or less of water, particularly 5 to 20% by mass of water is present with respect to the total mass of the reaction system.
  • Peptide hydrolase (C) is preferably at least one selected from the group consisting of protease, peptidase and hydrolase, particularly thermolysin. Such enzymes are readily available from Sigma-Aldrich Corporation. Peptide hydrolase (C) is preferably present in the reaction system in an amount of 1 to 4% by mass. By utilizing the positional specificity of the peptide hydrolase (C), the sequence of the constituent amino acids in the peptide that is the final reaction product can be easily controlled. Since water is substantially present in the reaction system, it is preferable to adjust the pH of the reaction system in the range of 4 to 10.5.
  • the reaction of the ionic liquidized first amino acid or peptide (A) with the second amino acid or peptide (B) is carried out by mixing them together at a temperature of 0 to 100 ° C., preferably room temperature (20 C.) to 70 ° C., preferably 30 to 40 ° C.
  • the reaction time is not particularly limited, and is usually 10 minutes to 3 days, preferably 30 minutes to 2 days, and more preferably 1 hour to 24 hours.
  • what can be used as an amino acid, a peptide, an ionic liquid, a preferable thing, a use ratio, etc. are the same as what was demonstrated about the said method.
  • the abundance ratio (molar ratio) of peptide and amino acid (including ionic liquid) contained in the liquid mixture containing peptide and amino acid is not particularly limited as long as the effect of the present invention is achieved.
  • the peptide: amino acid can be 99: 1 to 1:99, preferably 50: 1 to 1:50, more preferably 30: 1 to 1:30, and more preferably 20: 1 to 1:20.
  • the liquid mixture undergoes a condensation reaction between an amino group of an ionic liquid (ionic liquid B) having an amino acid as an anion and (3) an ester group of a peptide ester and / or an amino acid ester.
  • a preferable ratio can be 1:20 to 1: 1, preferably 1:10 to 1: 2, with peptide: amino acid depending on the reaction conditions.
  • a liquid comprising a first amino acid or peptide and a second amino acid or peptide different from the first amino acid or peptide, wherein at least one of the first amino acid or peptide and the second amino acid or peptide is
  • the liquid present as an ionic liquid that forms an ionic bond with a cation as an anion is acid-treated, and either one of the first amino acid or peptide and the second amino acid or peptide is preferentially precipitated and separated.
  • the liquid present as an ionic liquid that forms an ionic bond with a cation as an anion is acid-treated, and either one of the first amino acid or peptide and the second amino acid or peptide is preferentially precipitated and separated.
  • the liquid is preferably present as an ionic liquid in which both the first amino acid or peptide and the second amino acid or peptide form an ionic bond with a cation as an anion. That is, the liquid preferably includes an ionic liquid having a first amino acid or peptide as an anion and an ionic liquid having a second amino acid or peptide as an anion.
  • the liquid contains the first peptide and the second peptide
  • each of the first peptide and the second peptide for example, acid-treats the reaction solution obtained by the condensation reaction described in WO2012 / 014809, It can be obtained by selectively precipitating and separating the peptide which is the reaction product.
  • a condensation reaction between an amino group of an ionic liquid having a dipeptide as an anion and an ester group of an amino acid ester for example, a condensation reaction between an amino group of an ionic liquid having a dipeptide as an anion and an ester group of an amino acid ester
  • the reaction solution in which the amide bond is formed to synthesize the tripeptide is acid-treated, and the dipeptide or tripeptide is preferentially precipitated and separated from the reaction product, the amino group of the ionic liquid having the dipeptide as an anion, and the dipeptide
  • Examples include an embodiment in which a reaction solution in which an amide bond is formed by condensation reaction with an ester group of an ester to synthesize a tetrapeptide is subjected to an acid treatment, and the dipeptide or tetrapeptide is preferentially precipitated and separated from the reaction product.
  • Preferentially precipitating and separating any one of the first amino acid or peptide and the second amino acid or peptide means at least any of the solid first amino acid or peptide and second amino acid or peptide obtained by precipitation separation. It means that the ratio of the molar ratio or the mass ratio (preferably the molar ratio) between one (a) and the other amino acid or peptide (b) is larger in the above (a).
  • the molar ratio of (a) :( b) is greater than 1: 1, more preferably 2: 1 or more, more preferably 5: 1 or more, more preferably 10: 1 or more, more preferably 20 : 1 or more.
  • the liquid preferably contains an organic solvent.
  • an acid, an organic solvent, a cation, and other conditions the thing similar to the above is employable, for example.
  • the present invention can preferentially precipitate one amino acid or peptide without using the difference in solubility, so that even two types of amino acids and / or peptides having similar solubility can be separated. be able to.
  • the amino acids and peptides obtained by precipitation separation from the liquid mixture by the method of the present invention are usually obtained in the form of a free form (free form), but may be obtained in the form of a salt.
  • Peptides and amino acids obtained by precipitation separation in the method of the present invention are assumed to contain a salt form as described above. Examples of the salt include acid addition salts with acids used for acid treatment.
  • Peptides (oligopeptides and polypeptides) obtained by the method of the present invention can be used as active ingredients of pharmaceuticals, foods containing functional foods and seasonings, nutritional compositions such as infusions and feeds, active ingredients of pharmaceuticals, It can be used widely as an active ingredient of various reagents.
  • the present invention will be specifically described with reference to examples.
  • Example 1 Dipeptide [component (D)] produced by reaction of glycine methyl ester hydrochloride [component (B)] with glycine ionic liquid (GlyPBu 4 ) [component (A)], unreacted component (A) Isolation of glycine and tetrabutylphosphonium chloride The following reaction was carried out by the following procedure. The structures and abbreviations of the compounds used are shown below. PBu4 represents a tetrabutylphosphonium ion.
  • Example 2 Peptide [component (D)] produced by reaction of glycine methyl ester hydrochloride [component (B)] with alanine ionic liquid (AlaPBu 4 ) [component (A)], unreacted component (A) Isolation of alanine and tetrabutylphosphonium chloride in the following reaction was carried out by the following procedure. The structures and abbreviations of the compounds used are shown below. PBu 4 represents a tetrabutylphosphonium ion.
  • Example 2 In the same manner as in Example 1 except that an alanine ionic liquid (AlaPBu 4 ) [component (A)] was used instead of glycine ionic liquid (GlyPBu 4 ) [component (A)], the reaction and components were separated. went. Operation charts and results in Example 2 are shown below.
  • Example 3 Peptide [component (D)] produced by the reaction of glycine methyl ester hydrochloride [component (B)] and [emim] [Gly] [component (A)], unreacted component (A) Isolation of glycine and 1-ethyl-3-methylimidazolium chloride The following reaction was carried out by the following procedure. The structures and abbreviations of the compounds used are shown below.
  • [Emim] is 1-ethyl-3-methylimidazolium ion.
  • the reaction and components were separated in the same manner as in Example 1 except that [emim] [Gly] [component (A)] was used instead of GlyPBu 4 [component (A)]. Operation charts and results in Example 3 are shown below.
  • Example 4 Peptide [component (D)] produced by reaction of glycine methyl ester hydrochloride [component (B)] with [TBA] [Gly] [component (A)], unreacted component (A) Isolation of glycine and tetrabutylammonium chloride
  • the following reaction was carried out by the following operation. The structures and abbreviations of the compounds used are shown below.
  • [TBA] and NBu 4 are tetrabutylammonium ions.
  • the reaction and components were separated in the same manner as in Example 1 except that [TBA] [Gly] [component (A)] was used instead of GlyPBu 4 [component (A)]. Operation charts and results in Example 4 are shown below.
  • Example 5 Dipeptide [component (D)] produced by reaction of glycine methyl ester hydrochloride [component (B)] with glycine ionic liquid (GlyPBu 4 ) [component (A)], unreacted component (A) Isolation of Glycine and Tetrabutylphosphonium Chloride
  • glycine methyl ester hydrochloride component (B)
  • GlyPBu 4 glycine ionic liquid
  • A unreacted component
  • Isolation of Glycine and Tetrabutylphosphonium Chloride The reaction and components were separated in the same manner as in Example 1 except that acetonitrile (CH 3 CN) was used instead of chloroform as the organic solvent. Operation charts and results in Example 5 are shown below.
  • Example 6 Dipeptide [component (D)] produced by reaction of glycine methyl ester hydrochloride [component (B)] with glycine ionic liquid (GlyPBu 4 ) [component (A)], unreacted component (A) Isolation of glycine and tetrabutylphosphonium sulfate in the same manner as in Example 1 except that sulfuric acid was used instead of the combination of TMSCl and MeOH, which is a compound that generates an acid by reaction. Separation was performed. Operation charts and results in Example 6 are shown below.
  • Example 7 Dipeptide [component (D)] produced by reaction of glycine methyl ester hydrochloride [component (B)] with glycine ionic liquid (GlyPBu 4 ) [component (A)], unreacted component (A) Isolation of Glycine and Tetrabutylphosphonium Chloride
  • THF tetrahydrofuran
  • Example 8 Separation of glycine, glycylglycine and tetrabutylphosphonium acetate from a reaction mixture in which glycylglycine was synthesized by reaction of glycine ionic liquid (Gly-PBu4) and glycine methyl ester hydrochloride Glycine ionic liquid (glycine tetrabutyl Glycine methyl ester hydrochloride (125.5 mg, 1.0 mmol) was added to phosphonium salt: 1.78 g, 5.35 mmol), and the atmosphere was replaced with argon, followed by heating and stirring at 60 ° C. for 3 hours.
  • Glycine ionic liquid glycine tetrabutyl Glycine methyl ester hydrochloride
  • Filtrate residue (1.99 g): Tetrabutylphosphonium, glycylglycine and acetic acid signals were confirmed. A small amount of glycine ionic liquid was also detected. Furthermore, heavy water was removed from the filtrate residue solution after analysis by distillation under reduced pressure, and chloroform (6 mL) and acetic acid (170 mg, 2.8 mmol) were added at room temperature. The precipitated white solid was separated by filtration. The solvent contained in the filtrate was removed by distillation under reduced pressure, and proton NMR analysis was performed. Solid (87.4 mg): Glycine and glycylglycine signals were detected at a ratio of 1:24. From the weight and signal ratio, the yield of glycylglycine was calculated to be 65%. Filtrate residue (2.00 g): Tetrabutylphosphonium and acetic acid signals were confirmed.
  • Example 9 Separation of alanyl glycine from a reaction mixture in which alanyl glycine was synthesized by reaction of glycine ionic liquid (Gly-PBu4) and alanine methyl ester hydrochloride
  • glycine ionic liquid glycine tetra Butylphosphonium salt
  • alanine methyl ester hydrochloride was reacted and worked up.
  • the acetic acid used for crystallization was 3.2 mmol for the first time and 2.8 mmol for the second time.
  • Example 10 Separation of valylglycine from a reaction mixture in which valylglycine was synthesized by reaction of glycine ionic liquid (Gly-PBu4) and valine methyl ester hydrochloride
  • glycine ionic liquid glycine tetrabutylphosphonium
  • valine methyl ester hydrochloride glycine ionic liquid (glycine tetrabutylphosphonium) Salt) and valine methyl ester hydrochloride were reacted.
  • the acetic acid used for crystallization was 3.2 mmol for the first time and 4.7 mmol for the second time.
  • the molar ratio of glycine to valylglycine in the solid obtained by the second crystallization was 1: 5, and the yield of valylglycine was 25%.
  • Example 11 Separation of alanylalanine from a reaction mixture in which alanylalanine was synthesized by reaction of alanine ionic liquid (Ala-PBu4) and alanine methyl ester hydrochloride
  • alanine ionic liquid alanine tetra Butylphosphonium salt
  • alanine methyl ester hydrochloride were reacted and worked up.
  • the acetic acid used for crystallization was 3.2 mmol for the first time and 4.8 mmol for the second time.
  • the molar ratio of alanine to alanylalanine in the solid obtained by the second crystallization was 1:15, and the yield of alanylalanine was 67%.
  • Example 12 Separation of alanyl valine from a reaction mixture obtained by synthesizing alanyl valine by reaction of valine ionic liquid (Val-PBu4) and alanine methyl ester hydrochloride. And alanine methyl ester hydrochloride were reacted and worked up.
  • the acetic acid used for crystallization was 3.2 mmol for the first time and 2.8 mmol for the second time.
  • the molar ratio of valine to alanyl valine in the solid obtained by the second crystallization was 1: 6, and the yield of alanyl valine was 55%.
  • the results of Examples 8 to 12 are summarized in the following table.
  • Gly-PBu4 Glycine tetrabutylphosphonium salt
  • Ala-PBu4 Alanine tetrabutylphosphonium salt
  • Val-PBu4 Valine tetrabutylphosphonium salt
  • Example 13 Separation of glycylalanine from a reaction mixture in which glycylalanine was synthesized by reaction of alanine ionic liquid (Ala-PBu4) and glycine methyl ester hydrochloride
  • alanine ionic liquid alanine tetra Butylphosphonium salt
  • glycine methyl ester hydrochloride were reacted and worked up.
  • the acetic acid used for crystallization was 3.2 mmol for the first time and 6.7 mmol for the second time.
  • the molar ratio of alanine to glycylalanine in the solid obtained by the second crystallization was 1:24, and the yield of glycylalanine was 62%.
  • Example 14 Separation of prolyl glycine from a reaction mixture obtained by synthesizing prolyl glycine by reaction of glycine ionic liquid (Gly-PBu4) and proline methyl ester hydrochloride
  • glycine ionic liquid glycine tetra Butylphosphonium salt
  • proline methyl ester hydrochloride glycine tetra Butylphosphonium salt
  • the acetic acid used for crystallization was 3.3 mmol for the first time and 2.9 mmol for the second time.
  • the molar ratio of glycine to prolyl glycine in the solid obtained by the second crystallization was 1:85, and the yield of prolyl glycine was 76%.
  • Example 15 Separation of valylalanine from a reaction mixture obtained by synthesizing valylalanine by reaction of alanine ionic liquid (AlaPBu4) and valine methyl ester hydrochloride
  • alanine ionic liquid glycine tetrabutylphosphonium salt
  • valine methyl ester hydrochloride were reacted and worked up.
  • the acetic acid used for crystallization was 3.2 mmol for the first time and 2.8 mmol for the second time.
  • the molar ratio of alanine to valylalanine in the solid obtained by the second crystallization was 1:10, and the yield of valylalanine was 36%.
  • Example 16 Separation of glycylphenylalanine from a reaction mixture in which glycylphenylalanine was synthesized by reaction of phenylalanine ionic liquid (PhePBu4) and glycine methyl ester hydrochloride
  • phenylalanine ionic liquid phenylalanine tetrabutylphosphonium) Salt
  • glycine methyl ester hydrochloride were reacted.
  • the acetic acid used for crystallization was 3.2 mmol for the first time and 4.6 mmol for the second time.
  • the molar ratio of phenylalanine to glycylphenylalanine in the solid obtained by the second crystallization was 1: 1.2, and the yield of glycylphenylalanine was 55%.
  • Example 17 Separation of alanylphenylalanine from a reaction mixture in which alanylphenylalanine was synthesized by reaction of phenylalanine ionic liquid (PhePBu4) and alanine methyl ester hydrochloride
  • phenylalanine ionic liquid phenylalanine tetrabutylphosphonium
  • alanine methyl ester hydrochloride was reacted and worked up.
  • the acetic acid used for crystallization was 3.2 mmol for the first time and 4.9 mmol for the second time.
  • the molar ratio of phenylalanine to alanylphenylalanine in the solid obtained by the second crystallization was 1: 1, and the yield of alanylphenylalanine was 62%.
  • Example 18 Separation of glycine, glycylglycine and tetrabutylphosphonium chloride from a mixture of glycine ionic liquid (Gly-PBu4) and glycylglycine ionic liquid (Gly-Gly-PBu4) (molar ratio 4.5: 1) Glycine ionic liquid (glycine tetrabutylphosphonium salt: 3.00 g, 9.00 mmol;) and glycylglycine ionic liquid (glycylglycine tetrabutylphosphonium salt: purity 86.7%, 0.91 g, 2.02 mmol) mixed And a uniform liquid was obtained.
  • Glycine ionic liquid glycine tetrabutylphosphonium salt: 3.00 g, 9.00 mmol
  • glycylglycine ionic liquid glycylglycine tetrabutylphosphonium salt: purity
  • Example 19 Separation of glycine, glycylglycine and tetrabutylphosphonium chloride from a mixture (molar ratio 1: 1) of glycine ionic liquid (Gly-PBu4) and glycylglycine ionic liquid (Gly-Gly-PBu4) Liquid (glycine tetrabutylphosphonium salt: 2.00 g, 6.00 mmol;) and glycylglycine ionic liquid (glycylglycine tetrabutylphosphonium salt: purity 86.7%, 2.70 g, 6.00 mmol) A uniform solution was obtained.
  • Example 20 Separation of glycine, glycylglycine and tetrabutylphosphonium chloride from a mixture of glycine ionic liquid (Gly-PBu4) and glycylglycine ionic liquid (Gly-Gly-PBu4) (molar ratio 1: 4.5) Glycine ionic liquid (glycine tetrabutylphosphonium salt: 0.576 g, 1.73 mmol;) and glycylglycine ionic liquid (glycylglycine tetrabutylphosphonium salt: purity 86.7%, 3.46 g, 7.68 mmol) mixed And a uniform liquid was obtained.
  • Glycine ionic liquid glycine tetrabutylphosphonium salt: 0.576 g, 1.73 mmol
  • glycylglycine ionic liquid glycylglycine tetrabutylphosphonium salt
  • Example 21 Separation of glycine, glycylglycine and tetrabutylphosphonium chloride from a mixture of glycine ionic liquid (Gly-PBu4) and glycylglycine ionic liquid (Gly-Gly-PBu4) (molar ratio 4.5: 1) (Use ethyl acetate as the solvent to be added and hydrogen chloride as the acid to be added.) Glycine ionic liquid (glycine tetrabutylphosphonium salt: 3.00 g, 9.00 mmol;) and glycylglycine ionic liquid (glycylglycine tetrabutylphosphonium salt: purity 86.7%, 0.91 g, 2.02 mmol) mixed And a uniform liquid was obtained.
  • Glycine ionic liquid glycine tetrabutylphosphonium salt
  • Example 22 Glycylglycine and glycylglycyl from a mixture of glycylglycine ionic liquid (Gly-Gly-PBu4) and glycylglycylglycine ionic liquid (Gly-Gly-Gly-PBu4) (molar ratio 1: 1) Separation of glycine and tetrabutylphosphonium chloride Glycylglycine ionic liquid (glycylglycine tetrabutylphosphonium salt: 2.31 g, 5.1 mmol;) and glycylglycylglycine ionic liquid (glycylglycylglycine tetrabutylphosphonium salt: 2.
  • Example 23 Separation of tyrosylglycylglycine from a reaction mixture in which tyrosylglycylglycine was synthesized by reaction of glycylglycine ionic liquid (Gly-Gly-PBu4) with tyrosine methyl ester hydrochloride Glycylglycine ionic liquid (glycyl Tyrosine methyl ester hydrochloride (232.7 mg, 1.0 mmol) was added to glycine tetrabutylphosphonium salt (2.09 g, 5.34 mmol), followed by argon substitution, and the mixture was heated and stirred at 60 ° C. for 48 hours.
  • Glycylglycine ionic liquid Glycyl Tyrosine methyl ester hydrochloride
  • the precipitated white solid was separated by filtration, washed with chloroform (9 mL), and dried under reduced pressure at 40 ° C.
  • the obtained solid contained 0.316 g of glycylglycine and 0.033 g of tyrosylglycylglycine (weight ratio 91: 9, molar ratio 96: 4).
  • the purity as sylglycine was 77%, and the recovery rate was 50%.
  • the filtrate was dried under reduced pressure, chloroform (6 mL) was added, acetic acid (126.0 mg, 2.09 mmol) was added with cooling and stirring to 0 ° C., and the mixture was stirred at 0 ° C. for 30 min.
  • the precipitated white solid was separated by filtration, washed with chloroform (9 mL), and dried under reduced pressure at 40 ° C.
  • the obtained solid contained 0.095 g of glycylglycine and 0.014 g of tyrosylglycylglycine (weight ratio 87:13, molar ratio 85:15).
  • the purity as sylglycine was 83%, and the recovery rate was 22%.
  • the filtrate was dried under reduced pressure, chloroform (6 mL) was added, acetic acid (63.5 mg, 1.05 mmol) was added with cooling and stirring to 0 ° C., and the mixture was stirred at 0 ° C.
  • the extract was washed with chloroform (9 mL) and dried under reduced pressure at 40 ° C.
  • the obtained solid (0.003 g)
  • it contained 0.0012 g of glycylglycine and 0.0010 g of tyrosylglycylglycine (weight ratio 55:45, molar ratio 72:28).
  • the purity as sylglycine was 46%, and the recovery rate was 0.1%.
  • the filtrate was stored at room temperature for 1.5 days, and the precipitated white solid was separated by filtration, washed with chloroform (9 mL), and dried under reduced pressure at 40 ° C.

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Abstract

La présente invention concerne un procédé de production d'un peptide, caractérisé en ce qu'un mélange liquide contenant un peptide et un acide aminé est présent sous la forme d'un liquide ionique qui forme une liaison ionique avec un cation. Au moins un élément choisi parmi le peptide et l'acide aminé est un anion, et ce mélange liquide est soumis à un traitement acide, et le peptide est, de préférence, précipité et séparé, ou l'acide aminé est, de préférence, précipité et séparé, puis le produit obtenu est soumis à un autre traitement acide et le peptide est, de préférence, précipité et séparé.
PCT/JP2015/059317 2014-03-26 2015-03-26 Procédé de production de peptide WO2015147138A1 (fr)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008535483A (ja) * 2005-04-22 2008-09-04 ザ プロクター アンド ギャンブル カンパニー イオン性液体を使用するバイオマスからのバイオポリマーの抽出
JP2008537733A (ja) * 2005-03-15 2008-09-25 マクギル ユニバーシティー イオン液体担持合成
WO2012014809A1 (fr) * 2010-07-26 2012-02-02 味の素株式会社 Procédé de production d'un peptide
WO2012014808A1 (fr) * 2010-07-26 2012-02-02 味の素株式会社 Procédé de production d'un peptide

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008537733A (ja) * 2005-03-15 2008-09-25 マクギル ユニバーシティー イオン液体担持合成
JP2008535483A (ja) * 2005-04-22 2008-09-04 ザ プロクター アンド ギャンブル カンパニー イオン性液体を使用するバイオマスからのバイオポリマーの抽出
WO2012014809A1 (fr) * 2010-07-26 2012-02-02 味の素株式会社 Procédé de production d'un peptide
WO2012014808A1 (fr) * 2010-07-26 2012-02-02 味の素株式会社 Procédé de production d'un peptide

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
MIAO W. ET AL.: "Ionic-liquid-supported peptide synthesis demonstrated by the synthesis of leu5-enkephalin", J. ORG. CHEM., vol. 70, no. 8, 2005, pages 3251 - 3255, XP002409192 *
MIAO W. ET AL.: "Ionic-liquid-supported synthesis: a novel liquid-phase strategy for organic synthesis", ACC. CHEM. RES., vol. 39, no. 12, 2006, pages 897 - 908, XP055082093 *

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