WO2015045052A1 - Procédé pour l'introduction d'acide aminé contenant du fluor dans un peptide ou une protéine - Google Patents

Procédé pour l'introduction d'acide aminé contenant du fluor dans un peptide ou une protéine Download PDF

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WO2015045052A1
WO2015045052A1 PCT/JP2013/076025 JP2013076025W WO2015045052A1 WO 2015045052 A1 WO2015045052 A1 WO 2015045052A1 JP 2013076025 W JP2013076025 W JP 2013076025W WO 2015045052 A1 WO2015045052 A1 WO 2015045052A1
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amino acid
protein
peptide
fluorine
trna
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PCT/JP2013/076025
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Japanese (ja)
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秀和 ▲斉▼木
雪憲 長谷川
英一 小関
真清 瀧
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株式会社島津製作所
国立大学法人電気通信大学
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Priority to PCT/JP2013/076025 priority Critical patent/WO2015045052A1/fr
Priority to JP2015538697A priority patent/JP6099756B2/ja
Publication of WO2015045052A1 publication Critical patent/WO2015045052A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y203/00—Acyltransferases (2.3)
    • C12Y203/02—Aminoacyltransferases (2.3.2)
    • C12Y203/02006—Leucyltransferase (2.3.2.6)

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  • the present invention relates to a technique applied to biodistribution measurement of PET diagnostic reagents, antibody-type diagnostic agents, peptide-type diagnostic agents, and protein-type pharmaceutical agents. More specifically, the present invention relates to a method for introducing a fluorine-containing amino acid into a peptide or protein (fluorine introduction method), particularly, a method for introducing an 18 F-containing amino acid into a peptide or protein ( 18 F labeling method).
  • the radioisotope 18 F (half-life: 110 minutes) of fluorine is known in the clinical field as the most useful nuclide for PET probe applications.
  • [ 18 F] FDG which is a glucose derivative
  • This diagnostic agent uses the accumulation of [ 18 F] FDG in cancer tissue due to high sugar metabolic activity in cancer tissue, and is used for ultra-early diagnosis of cancer and judgment of therapeutic effect. .
  • Patent Document 1 JP-A-2009-106268 (Patent Document 1) describes FMT (fluoromethyl-L-tyrosine) and FET (fluoroethyl-L-tyrosine), which are fluorine-containing amino acids, in one-pot using an enzymatic method. A method of introducing into the N-terminus of a peptide or protein (Nexta method) is described.
  • [ 18 F] SFB N-succinimidyl-4- [ 18 F] fluorobenzoate
  • 18 F] SFB N-succinimidyl-4- [ 18 F] fluorobenzoate
  • the method of introducing an 18 F-labeled amino acid into a peptide or protein is preferable because it does not impair the function of the peptide or protein to be labeled.
  • Examples of the technique for introducing 18 F-labeled amino acid include peptide synthesis, genetic engineering, and enzymatic chemistry.
  • the peptide synthesis method has a problem that it takes one day or more to introduce one amino acid molecule, the genetic engineering method is troublesome, and the enzyme chemical method has a problem that the introduction efficiency is not so good.
  • the fluorine-containing amino acid to be introduced is an [ 18 F] -containing amino acid
  • there is a problem in using the conventional Nexta method That is, first, it takes about 20 to 30 minutes to prepare [ 18 F] -containing amino acid (synthesis, purification, radiochemical purity test, etc.). Subsequently, when the prepared [ 18 F] -containing amino acid is subjected to the conventional Nexta method, the Nexta method itself requires 20 minutes. Therefore, in order to introduce an 18 F-labeled amino acid into a peptide or protein, a total time of 40 to It takes about 50 minutes. In this case, it is necessary to reduce the time of the Nexta method, judging from the half-life of the radioisotope 18 F being 110 minutes.
  • an object of the present invention is to provide a method capable of introducing a fluorine-containing amino acid into a peptide or protein in a shorter time than a conventional method using an enzymatic chemical method.
  • an object of the present invention is to provide a method capable of introducing an [ 18 F] -containing amino acid into a peptide or protein in a shorter time than a conventional method using an enzymatic chemical method.
  • the present inventors have previously formed a complex of a peptide or protein into which a fluorine-containing amino acid is to be introduced and an aminoacyl-tRNA protein transferase, and then performing a Nexta-like reaction, It has been found that the time for introducing fluorine-containing amino acids can be greatly reduced, and the present invention has been completed.
  • the present invention includes the following inventions.
  • the peptide or protein is used in an amount of 2/1 to 10,000 / 1, expressed in molar ratio with respect to the aminoacyl tRNA protein transferase. ) Or the method according to (2).
  • the fluorine-containing amino acids are 2-amino-3- (4- [ 18 F] fluoromethoxyphenyl) propionic acid ([ 18 F] FMT) and 2-amino-3- [4- (2- [ 18
  • the N-terminal residue of the peptide or protein to which the fluorine-containing amino acid is to be introduced is a basic amino acid residue, and the fluorine-containing amino acid is introduced to the N-terminus of the peptide or protein, The method according to any one of (6).
  • peptide means an amino acid sequence containing two or more amino acids.
  • protein refers to a peptide having a molecular weight of 5,000 or more.
  • the protein in the present invention is a peptide that can exhibit some function / activity in vivo, for example, when it is produced in vivo.
  • amino acid includes ⁇ -, ⁇ -, and ⁇ -amino acids.
  • introducing fluorine is performed by introducing a fluorine-containing amino acid. Therefore, in this specification, “fluorine introduction” is used in the same meaning as the introduction of a fluorine-containing amino acid.
  • the following (2-1) to (2-10) may be used in order to more efficiently introduce a fluorine-containing amino acid into a peptide or protein using an enzymatic method. That is, in the above method, the efficiency of introducing the 18 F-labeled amino acid is improved by adding a specific sequence to the peptide or protein to which the 18 F-labeled amino acid is to be introduced in advance to obtain an activated peptide or protein. Introduction of the activation sequence is an optional step.
  • an activation sequence having at least an amino acid residue whose side chain is —H, —CH 3 or —C (CH 3 ) C 2 H 5 is added to a peptide or protein to which fluorine is to be introduced, and the activation peptide Or obtaining a protein; Mixing the obtained activated peptide or protein and aminoacyl-tRNA protein transferase to form a complex of the peptide or protein and the aminoacyl-tRNA protein transferase; Mixing the complex with a reagent solution containing a fluorine-containing amino acid to be introduced, tRNA, and aminoacyl-tRNA synthetase, and introducing the fluorine-containing amino acid into the peptide or protein.
  • the second amino acid residue from the N-terminus of the peptide or protein into which fluorine is to be introduced is an amino acid residue having a hydroxyl group in the side chain, a hydrophilic amino acid residue having a positive charge in the side chain, and a side chain of -H , —CH 3 , —C (CH 3 ) C 2 H 5 and —CH 2 CONH 2, which is not any of the amino acid residues, the method according to (2-2) above.
  • a complex of a peptide or protein to which a fluorine-containing amino acid is to be introduced and an aminoacyl-tRNA protein transferase is formed in advance, and then a reaction similar to the Nexta method is performed, whereby the reaction of introducing a fluorine-containing amino acid is performed.
  • Time can be greatly shortened (for example, time of 1 minute or less).
  • the method of the present invention is very useful for introducing [ 18 F] -containing amino acids into peptides or proteins, judging from the half-life of radioisotope 18 F being 110 minutes.
  • Example 1 the measurement result of fluorescence detection HPLC in the initial 0 minute, incubation 1 minute, and incubation 10 minutes is shown.
  • the peptide or protein into which fluorine is to be introduced does not matter whether it has a natural amino acid sequence or a non-natural amino acid sequence.
  • a peptide or protein having a natural amino acid sequence can be obtained, for example, by isolation from various living organisms. Alternatively, it may be artificially produced using a known technique such as a genetic engineering technique or organic synthesis.
  • a peptide or protein having an unnatural amino acid sequence can also be artificially produced using a known technique such as a genetic engineering technique or organic synthesis.
  • N-terminal residue is a basic amino acid residue (for example, lysine residue or arginine residue) for the progress of the Nexta method.
  • an activation sequence is added to the peptide or protein into which fluorine is to be introduced in advance to activate. It may be a peptide or a protein.
  • Activation sequence The peptide or protein into which fluorine is to be introduced is optionally converted into an activated peptide or protein by adding an activation sequence prior to complex formation (before fluorine introduction).
  • the activation sequence includes at least a peptide sequence consisting of specific 2 amino acid residues.
  • Each of the specific 2 amino acid residues may be a residue derived from a natural amino acid or a residue derived from an unnatural amino acid.
  • the N-terminal amino acid residue of the activation sequence is a basic amino acid. Specific examples include lysine residues and arginine residues.
  • amino acid residue adjacent to the N-terminal amino acid residue of the activation sequence examples include the following.
  • -Amino acid residue having a hydroxyl group in the side chain hydroxyl group includes normal hydroxyl group and phenolic hydroxyl group
  • a hydrophilic amino acid residue having a positive charge in the side chainAmino acid residue in which the side chain is —H, —CH 3 , —C (CH 3 ) C 2 H 5 or —CH 2 CONH 2
  • amino acid residues having a normal hydroxyl group (not a phenolic hydroxyl group) in the side chain and hydrophilicity having a positive charge in the side chain Amino acid residues are more preferred.
  • amino acids having a hydroxyl group in the side chain include serine, threonine, and tyrosine.
  • hydrophilic amino acids having a positive charge in the side chain include lysine and arginine.
  • An example of an amino acid whose side chain is —H is glycine, an example of an amino acid whose side chain is —CH 3 is alanine, and an example of an amino acid whose side chain is —C (CH 3 ) C 2 H 5 is isoleucine.
  • An example of an amino acid whose side chain is —CH 2 CONH 2 is asparagine.
  • the activation efficiency should be approximately in the order of serine, arginine, threonine, lysine, glycine, alanine, isoleucine, tyrosine and asparagine. Has been confirmed by the present inventors.
  • the activation sequence can include a spacer group attached to the second amino acid residue.
  • the spacer group can give the activation sequence a structure that preferably matches the substrate binding pocket of the enzyme used in the fluorine-containing amino acid introduction step described below.
  • the spacer group may be a peptide chain or a structure other than that.
  • structures other than peptides include divalent organic groups.
  • the divalent organic group include an alkylene oxide-containing group.
  • the alkylene oxide in the alkylene oxide-containing group may be an alkylene oxide having 2 to 6 carbon atoms, preferably ethylene oxide or propylene oxide.
  • the divalent organic group may be a polyalkylene oxide-containing group having these as a repeating unit.
  • the spacer group may be appropriately labeled with an isotope or a fluorescent label by those skilled in the art according to various measurement methods and detection methods combined with the present invention.
  • the activation sequence is optionally added to the peptide or protein into which fluorine is to be introduced.
  • the activation sequence is added to the N-terminus of the peptide or protein into which fluorine is to be introduced. Thereby, an activated peptide or protein is obtained.
  • the second amino acid residue is a peptide or protein into which fluorine is to be introduced.
  • the spacer group binds to the peptide or protein into which fluorine is to be introduced.
  • the method of adding the activation sequence can be appropriately selected by those skilled in the art according to the binding mode between the activation sequence and the peptide or protein into which fluorine is to be introduced.
  • a normal peptide synthesis method can be used. More specifically, there is a peptide solid phase synthesis method.
  • a polystyrene polymer gel bead with a diameter of about 0.1 mm whose surface is modified with an amino group is used as the solid phase, and amino acid chains are extended one by one through a dehydration reaction.
  • the sequence of the target peptide is completed, it is cut out from the solid surface to obtain the target substance.
  • it in order to change the N terminus of a protein to an arbitrary sequence, it can be rearranged into an arbitrary sequence using an engineered gene at the stage of protein synthesis using E. coli.
  • the binding mode is not a peptide bond (eg PEG chain)
  • an active ester group succinimide or the like at the end of the target linker and react with the N-terminal amino acid of the peptide / protein.
  • a peptide or protein into which a fluorine-containing amino acid is to be introduced and an aminoacyl-tRNA protein transferase are mixed to form a complex of the peptide or protein and the aminoacyl-tRNA protein transferase.
  • the activation sequence is arbitrarily introduced, the obtained activation peptide or protein and aminoacyl tRNA protein transferase are mixed to form a complex of the peptide or protein and the aminoacyl tRNA protein transferase.
  • the aminoacyl-tRNA protein synthase is an enzyme that transfers an amino acid added to the 3 ′ end of tRNA to the amino terminus (N-terminus) of the protein and catalyzes a peptide bond forming reaction without using the genetic code.
  • the aminoacyl tRNA protein transferase may be a wild type or a mutant type having the same function as the wild type.
  • the aminoacyl-tRNA protein transferase can be prepared by appropriately synthesizing by a person skilled in the art according to a known method (for example, a genetic engineering technique) or by purchasing a commercially available product.
  • L / F transferase leucyl / phenylalanyl tRNA protein transferase
  • L / F transferase is derived from E. coli and is an enzyme that catalyzes the reaction of transferring hydrophobic amino acids such as phenylalanine, leucine, and methionine bound to tRNA to peptides or proteins having lysine or arginine at the N-terminus. is there.
  • an excess molar amount of the peptide or protein is used with respect to the aminoacyl-tRNA protein transferase.
  • the peptide or protein (P) is expressed in molar ratio (P / T) to the aminoacyl tRNA protein transferase (T), for example, in an amount of 2/1 to 10,000 / 1.
  • the amount is preferably 10/1 to 2,000 / 1, more preferably 50/1 to 500/1.
  • the reaction cycle is efficiently performed, and the time for introducing the fluorine-containing amino acid into the complex is greatly reduced (for example, 1 minute) The following time) There is a great advantage when the fluorine-containing amino acid is an [ 18 F] -containing amino acid.
  • An amino acid having fluorine is a molecule in which fluorine is introduced into the basic skeleton of amino acid (a structure having an amino group and a carboxyl group in the same molecule).
  • a molecule in which fluorine is introduced into a natural or unnatural amino acid as a basic skeleton by fluorine substitution or by labeling with a fluorine-containing group can be mentioned.
  • the fluorine nuclide is not particularly limited, but the radioisotope 18 F is particularly preferable because it is useful in applications to PET probes.
  • An amino acid having fluorine is allowed to have a label or modification other than fluorine as appropriate.
  • labels include stable isotope labels and fluorescent labels. In this case, measurement and detection can be performed based on a mass value based on a stable isotope and a signal based on a fluorescent label.
  • modification include modification with an enzyme substrate and an antigenic substance. In this case, detection and purification can be performed based on an enzyme chemical technique or an enzyme immunoscience technique.
  • amino acids having fluorine include the following. 2-amino-3- (2,3,4,5,6-pentafluorophenyl) propionic acid (ie pentafluorophenylalanine) 2-amino-3- (4-fluoromethoxyphenyl) propionic acid (ie fluoromethyltyrosine) 2-amino-3- [4- (2-fluoroethoxy) phenyl] propionic acid (ie fluoroethyltyrosine)
  • 2-amino-3- (4-fluoromethoxyphenyl) propionic acid and 2-amino-3- [4- (2-fluoroethoxy) phenyl] propionic acid are preferred, and their radioactive properties
  • the isotopically labeled 2-amino-3- (4- [ 18 F] fluoromethoxyphenyl) propionic acid and 2-amino-3- [4- (2- [ 18 F] fluoroethoxy) phenyl] propionic acid More preferred.
  • the amino acid having fluorine is appropriately selected according to aminoacyl tRNA synthetase and aminoacyl tRNA protein transferase described later.
  • a fluorine-containing amino acid is introduced into the complex of the peptide or protein (or optionally activated peptide or protein) and the aminoacyl-tRNA protein transferase.
  • aaRS aminoacyl tRNA synthetase
  • tRNA can be converted to aminoacyl-tRNA by aminoacylation by aminoacyl-tRNA synthetase, and the aminoacyl group of aminoacyl-tRNA can be converted into a peptide or a peptide by aminoacyl-tRNA protein transferase (in the present invention, the complex). Any material can be used as long as it can be transferred to a protein, whether natural or non-natural. Therefore, it is not limited whether the tRNA structure has a cloverleaf secondary structure, and the size thereof is not limited.
  • the tRNA can be prepared by appropriate synthesis by a person skilled in the art according to a known method, or by purchasing a commercially available product. Specific examples of tRNA include tRNA Phe and tRNA Leu .
  • An aminoacyl-tRNA synthetase is an enzyme responsible for synthesizing an aminoacyl-tRNA that is generally a substrate for translation of the genetic code in a ribosome.
  • the aaRS may be a wild type or a mutant type.
  • the aaRS can be prepared by appropriately synthesizing by a person skilled in the art according to a known method (for example, genetic engineering technique), or by purchasing a commercially available product.
  • aaRS corresponding to lysine is called lysyl tRNA synthetase.
  • aaRS corresponding to the amino acid basic skeleton in the fluorine-containing amino acid to be introduced that is, having substrate specificity for the amino acid basic skeleton
  • an aaRS mutant in which a mutation is generated such that the substrate specificity for a fluorine-containing amino acid is higher than the substrate specificity for a fluorine-free amino acid is used.
  • E. coli-derived phenylalanyl tRNA synthetase mutant (Ala294 ⁇ Gly) (Chembiochem, 2002, 02-03, which has higher substrate specificity for fluoromethyltyrosine and fluoroethyltyrosine than that for phenylalanine. 235-237) can be used.
  • a buffer solution suitable for the introduction reaction of the fluorine-containing amino acid, salts, reducing agent, polyamine, energy source and the like can be appropriately contained.
  • the buffer include Hepes buffer and Tris-acetic acid.
  • the salts include magnesium salts and potassium salts.
  • the reducing agent include DTT (dithiothreitol).
  • the polyamine include spermidine.
  • the energy source include adenosine triphosphate.
  • the composition at the final concentration is 10 mM MgCl 2 , 1 mM Spermidine, 50 mM Hepes buffer (pH 7.6), and the composition at the final concentration is 2.5 mM ATP, 20 mM.
  • KCl, 2 mM DTT B solution can be mixed and used.
  • the concentration of each component in the mixed solution of the peptide or protein (or optionally activated peptide or protein) and the aminoacyl-tRNA protein transferase complex and the reagent solution can be arbitrarily set. it can.
  • the molar ratio of the peptide or protein complex to tRNA in the mixed solution is 30: 1 to 1: 1, more preferably 10: 1 to 4: 1.
  • the amount used may be small.
  • the molar ratio of the peptide or protein complex and aminoacyl-tRNA synthetase in the mixed solution is 100: 1 to 2: 1, more preferably 50: 1 to 20: 1.
  • the molar ratio of the peptide or protein complex to the fluorine-containing amino acid in the mixed solution is 100: 1 to 1: 1000, more preferably 25: 1 to 1: 400, still more preferably 1: 1 to 1: 120. It is. While the fluorine-containing amino acid can be introduced even in a small amount compared to the peptide or protein complex, it can inhibit the reaction system even in an excessive amount compared to the peptide or protein complex. It is not considered. Therefore, when using a valuable fluorine-containing amino acid such as a radioisotope fluorine-containing amino acid, the amount of fluorine-containing amino acid used can be kept small compared to the peptide or protein complex.
  • the amount of the activated peptide or protein complex used can be reduced to a small amount with respect to the fluorine-containing amino acid. Since a complex of the peptide or protein (optionally activated peptide or protein) and the aminoacyl-tRNA protein transferase is formed in advance, the reaction of introducing a fluorine-containing amino acid into the complex is very fast.
  • reaction conditions such as temperature and pH can be selected according to the enzyme used.
  • the reaction temperature is preferably 0 ° C to 50 ° C, more preferably 4 ° C to 37 ° C.
  • the reaction pH is preferably 6-9, more preferably 7-8.
  • a reaction time of about 10 minutes is sufficient, and the reaction is completed in about 1 minute.
  • the fluorine-containing amino acid is an [ 18 F] -containing amino acid.
  • the fluorine-containing amino acid to be introduced tRNA, and aminoacyl
  • a reaction solution in which a reagent solution containing tRNA synthetase is mixed is prepared.
  • Each component may be mixed at once or in any order.
  • the fluorine-containing amino acid can use 1 type or multiple types.
  • the reaction solution is subjected to incubation (for example, 37 ° C., 60 minutes), whereby the fluorine-containing amino acid introduction reaction proceeds.
  • the fluorine-containing amino acid is an [ 18 F] -containing amino acid
  • the incubation time should be a short time, for example, about 10 minutes or less, preferably about 5 minutes, more preferably about 1 minute.
  • the reaction can be stopped by, for example, mixing a TFA (trifluoroacetic acid) aqueous solution.
  • TFA trifluoroacetic acid
  • the peptide or protein into which the fluorine-containing amino acid has been introduced is recovered as a target product by concentrated desalting.
  • the purified target product can be purified by concentration desalting by ZipTip treatment.
  • kits which can be used for the method of introduce
  • Items include at least tRNA, aminoacyl tRNA synthetase, and aminoacyl tRNA protein transferase. Each item may be provided in a state where it exists in a separate container, or for example, it may be provided in a state where tRNA and aminoacyl tRNA synthetase coexist in one container.
  • the aminoacyl tRNA protein transferase is present in the container by itself.
  • one or more fluorine-containing amino acids for example, phenylalanine and derivatives thereof
  • the above-described buffer solution, salts, reducing agent, polyamine, energy source, and the like may be included.
  • Lys-Bradykinin was used as a peptide into which a fluorine-containing amino acid was to be introduced.
  • fluorine-containing amino acid cold 2-amino-3- [4- (2-fluoroethoxy) phenyl] propionic acid (fluoroethyltyrosine: FET) was used.
  • Leucyl / phenylalanyl tRNA protein transferase (L / FT) was used as the aminoacyl tRNA protein transferase.
  • composition at the final concentration in the amino acid introduction step is MgCl 2 10 mM Supermidine 1 mM Hepes buffer (pH 7.6) 50 mM L / FT 2 ⁇ M ea Lys-Bradykinin 0.17 mM ea Then, the mixture was incubated at 37 ° C. for 5 minutes to convert all L / FT molecules into L / FT / Lys-Bradykinin complexes (complex solution A).
  • composition at the final concentration in the amino acid introduction step is MgCl 2 10 mM Supermidine 1 mM Hepes buffer (pH 7.6) 50 mM tRNA 6 ⁇ M FET 2 ⁇ M ARS 2 ⁇ M Then, the mixture was incubated at 37 ° C. for 10 minutes to form aminoacyl tRNA (reagent solution B).
  • Reagent solution B was added to complex solution A and incubated at 37 ° C. for 10 minutes to introduce FETs into Lys-Bradykinin.
  • reaction efficiency was calculated by fluorescence detection HPLC (manufactured by JASCO Corporation, PU2085) at each time point of the initial reaction (0 minutes, when mixing the complex solution A and the reagent solution B), 1 minute of reaction, and 10 minutes of reaction.
  • Fluorescence detection conditions ex. 274 nm em. 300nm (The reaction efficiency was calculated by detecting the decrease in FET over time.) ⁇ Development conditions: 0.1% TFA (0-2 min) 0.1% TFA-AN (0-100%; 2-12 min)

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Abstract

L'invention porte sur un procédé qui utilise un procédé chimique enzymatique et qui peut introduire un acide aminé contenant du fluor dans un peptide ou une protéine sur un temps plus court qu'à l'aide des procédés classiques. Le procédé pour l'introduction d'un acide aminé contenant du fluor dans un peptide ou une protéine selon l'invention comprend : un processus consistant à mélanger le peptide ou la protéine dans lequel l'acide aminé contenant du fluor doit être introduit avec une aminoacyl-ARNt-protéine transférase pour former un complexe dudit peptide ou de ladite protéine et de ladite aminoacyl-ARNt-protéine transférase ; et un processus consistant à mélanger ledit complexe avec l'acide aminé contenant du fluor devant être introduit et une solution de réactifs contenant de l'ARNt et une aminoacyl-ARNt synthase pour introduire l'acide aminé contenant du fluor dans le peptide ou la protéine.
PCT/JP2013/076025 2013-09-26 2013-09-26 Procédé pour l'introduction d'acide aminé contenant du fluor dans un peptide ou une protéine WO2015045052A1 (fr)

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CN110483278A (zh) * 2019-08-06 2019-11-22 唐刚华 2,2-二氟-3-18f-氟代丙酸及其合成方法与应用
JP2021515561A (ja) * 2018-03-08 2021-06-24 ザ・リージエンツ・オブ・ザ・ユニバーシテイー・オブ・カリフオルニア 生体反応性組成物およびそれらの使用方法

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JP2021515561A (ja) * 2018-03-08 2021-06-24 ザ・リージエンツ・オブ・ザ・ユニバーシテイー・オブ・カリフオルニア 生体反応性組成物およびそれらの使用方法
CN110483278A (zh) * 2019-08-06 2019-11-22 唐刚华 2,2-二氟-3-18f-氟代丙酸及其合成方法与应用

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