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WO2007114307A1 - Glycopeptide de type mucine comportant un squelette de polylactosamine - Google Patents

Glycopeptide de type mucine comportant un squelette de polylactosamine Download PDF

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Publication number
WO2007114307A1
WO2007114307A1 PCT/JP2007/057001 JP2007057001W WO2007114307A1 WO 2007114307 A1 WO2007114307 A1 WO 2007114307A1 JP 2007057001 W JP2007057001 W JP 2007057001W WO 2007114307 A1 WO2007114307 A1 WO 2007114307A1
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Prior art keywords
group
bond
sugar
residue
compound
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PCT/JP2007/057001
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English (en)
Japanese (ja)
Inventor
Shinichiro Nishimura
Masataka Fumoto
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National Institute of Advanced Industrial Science and Technology AIST
Shionogi and Co Ltd
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National Institute of Advanced Industrial Science and Technology AIST
Shionogi and Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4727—Mucins, e.g. human intestinal mucin
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K9/00—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof
    • C07K9/001—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof the peptide sequence having less than 12 amino acids and not being part of a ring structure
    • C07K9/005—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof the peptide sequence having less than 12 amino acids and not being part of a ring structure containing within the molecule the substructure with m, n > 0 and m+n > 0, A, B, D, E being heteroatoms; X being a bond or a chain, e.g. muramylpeptides

Definitions

  • the present invention relates to a novel compound useful as a primer for producing a glycopeptide, and a method for producing a glycopeptide using the primer.
  • the present invention also relates to glycopeptides obtained by the production method.
  • a sugar chain is a major component of a living body along with nucleic acids and proteins, and is a well-known power source for living bodies.
  • in vivo information transmission, protein quality control, and structural stabilization It has become clear that it has various higher-order functions such as labels for protein transport.
  • sugar chains compared to nucleic acids and proteins, sugar chains have not been established as a general preparation method, and the functions of sugar chains often function as complex carbohydrates bound to lipids and proteins. For this reason, there are a lot of unexplained studies on functions including structural information.
  • the ability to look at many things that seem to perform their functions together with sugar chains is very difficult to study in detail.
  • the main reason for this is that the preparation of the sugar amino acid as a raw material is complicated and it is difficult to prepare sugar amino acids having various sugar chain structures, and sugar amino acids having a large sugar chain structure have large steric hindrance.
  • the yield and reaction rate are slow, and it is also important to extend the sugar chain by chemical synthesis after glycopeptide construction. Position ⁇
  • the power of 3D control is difficult. In other words, with the current technology, the reaction yield is low and the time required for preparation is long.
  • it is difficult to prepare the synthetic raw material itself for glycopeptide synthesis it is custom-made to rapidly prepare the necessary sugar chain structure. It is extremely difficult to construct a glycopeptide library containing a complex sugar chain structure that is required for production and comprehensive functional analysis of glycopeptides and glycoproteins.
  • glycopeptides are synthesized by Fmoc amino acid (amino acid with amino group protected with 9 fluorenylmethyloxycarboxyl group, hereinafter 9 fluorenylmethyloxycarboxyl group is abbreviated as Fmoc).
  • Fmoc glycosylamino acid the basic peptide part is synthesized on a solid phase carrier by an automatic peptide synthesizer, the peptide part is released from the solid phase carrier, and once purified, it is purified organically or enzymatically.
  • a method of extending sugar chains one by one using a simple synthesis method is used. For this reason, a complicated operation and a long time are required for sugar chain elongation.
  • C. — H. Wong et al. Used a glycopeptide linked to amino-silica as a primer, extended the sugar chain using glycosyltransferase, and then hydrolyzed oc-chymotrypsin.
  • the peptide chain of the resulting glycopeptide is as short as Asn (asparagine) Gly (glycine) Phe (furalanine).
  • the yield of the sugar chain elongation reaction by glycosyltransferase is 55 to 65%, which is very sufficient. Not a thing.
  • the glycopeptide chain obtained by this method has 8 amino acid residues and has a sufficient length as a peptide chain, but the obtained glycopeptide was first introduced into a solid support.
  • the yield based on amino acids is less than 10%, which is not sufficient.
  • impurities such as unreacted substances accumulate through peptide synthesis and sugar chain synthesis, so that isolation and purification of the target product becomes difficult if the peptide chain and sugar chain structure are complex.
  • automatic peptide synthesis is usually carried out in an organic solvent, and glycosylation by glycosyltransferase is usually carried out in an aqueous solution, and the properties of the carrier required for each reaction are different. Even automatic synthesis is difficult.
  • the peptide chain of the glycopeptide obtained by this method is Asn (asparagine) -Gly (glycine), which is too short to be called a glycopeptide.
  • the C-terminal glycine residue is a glycinamide residue, and in some cases, it is necessary to convert the glycinamide residue to a glycine residue.
  • a glycosyltransferase sugar receptor is bound to a solid phase carrier, this is used as an affinity adsorbent, and a tissue extract containing a glycosyltransferase that can bind to this sugar receptor is contacted.
  • the glycosyltransferase is bound to the affinity adsorbent.
  • the affinity adsorbent to which the glycosyltransferase is bound is brought into contact with a solution containing a sugar nucleotide that can be used as a sugar donor by the glycosyltransferase, thereby releasing the glycosyltransferase and the sugar adsorbent.
  • a tissue extract containing a glycosyltransferase capable of binding to a sugar receptor in which one sugar residue is extended is brought into contact, and the same process is repeated to synthesize a desired sugar chain on a solid support. It is.
  • there is no specific data showing the usefulness of this method or its application to the synthesis of non-natural glycopeptides and a method for releasing the resulting sugar chain on the solid phase carrier is also disclosed. It has not been.
  • a useful polymerizable aromatic amino acid derivative is disclosed (see Patent Document 2).
  • this method involves complicated operations such as column purification and polymerization after peptide synthesis, since the peptide having a sugar residue is radically polymerized, and thus is weak in radicals and difficult to prepare glycopeptides containing sulfur atoms. The problem is that it takes time to switch to a sugar chain elongation reaction by an enzyme. The title is left.
  • Mucin is the main glycoprotein of mucus that covers the lumen of the gastrointestinal tract such as the trachea, gastrointestinal tract, and gonads.
  • MUC1 is a membrane-bound glycoprotein of epithelial cells and the first mucin that has been studied in detail.
  • MUC1 is a tandem repeat (HGVTSAPDTRPAPGSTA PPA (SEQ ID NO: 41)), a repetitive amino acid sequence containing serine and threonine that can be attached to an O-linked sugar chain. is there. Since sugar chain attachments do not occur in all serines and threonines, the degree of sugar chain extension varies, so there are many glycoproteins with the same amino acid sequence but different functions. Yes.
  • Non-patent Document 9 Nakamori, S .; Ota, DM; Karen, R .; Shirotani, K .; Irimura , T. Gastroenterology, 1994, 106, 353—361.
  • MU C1 degree of glycosylation of MU C1 (where sugar chains are introduced) and the structure of sugar chains differ between those derived from normal epithelium and those derived from cancer cells
  • Non-patent Document 10 Lloyd, K.
  • Tn which is a cancer-related sugar chain antigen
  • a mother nucleus structure such as ⁇ ⁇ , and sialyl ⁇ , sialyl ⁇ ⁇ , and sialyl Lewis ⁇ antigen and sialyl Lewis X antigen combined with sialic acid
  • Non-patent Document 12 Koganty, RR; Reddish, MR; Longenecker , BM Drug Discov. Today, 1996, 1, 190-198 .;
  • Biomira-Merck is developing a synthetic MUC 1 peptide vaccine that incorporates a 25-amino acid sequence of MUC1 cancer mucin in a liposomal formulation: “L BLP25”. Phasell targets lung cancer and prostate cancer. A clinical trial is ongoing.
  • Bio mira Merck has developed KLH (Keyhole limp et hemocyanin) that stimulates antibody production and T-cell responses to STn (disaccharide) specifically expressed in mucin on cancer cells.
  • KLH Keyhole limp et hemocyanin
  • STn disaccharide
  • Nishimura et al. Developed a water-soluble polymer primer for synthesizing glycopeptides (Patent Document 3).
  • Patent Document 3 a glycopeptide derivative having an aldehyde group or a ketone group at the end and containing a photocleavable linker for peptide solid phase synthesis is converted into a polymer primer, which is used for sugar conversion. The production of peptides is described.
  • Nishimura et al. Also developed a method for synthesizing mucin-type peptides and a glycopeptide related to MUC1 (Patent Document 4).
  • Patent Document 4 describes that a glycopeptide derivative having an aldehyde group or a ketone group at the end and containing an amino acid residue that can be cleaved by a protease is converted into a primer, and a glycopeptide is produced using this.
  • Patent Document 1 Japanese Patent Publication No. 5-500905
  • Patent Document 2 JP 2001-220399 A
  • Patent Document 3 International Publication No. 2005Z108417 Pamphlet
  • Patent Document 4 Pamphlet of International Publication No. 2006Z030840
  • Non-patent literature l Carbohvdr. Res., 124, 23 (1983)
  • Non-Patent Document 2 Carbohydr. Res., 228, 255 (1992)
  • Non-Patent Document 3 React. Polym., 22, 171 (1994)
  • Non-Patent Document 4 Carbohydr. Res., 265, 161 (1994)
  • Non-Patent Document 5 J. Am. Chem. Soc., 116, 1136 (1994)
  • Non-Patent Document 6 J. Am. Chem. Soc., 116, 11315 (1994)
  • Non-Patent Document 7 J. Am. Chem. Soc., 119, 8766 (1997)
  • Non-Patent Document 8 J. Chem. Soc. Chem. Commun., 1849 (1994)
  • Non-Patent Document 9 Gastroenterology, 106, 353-361 (1994)
  • Non-Patent Document 10 Biol. Chem., 271, 33325-33334 (1996)
  • Non-Patent Document ll Glycobiology, 10, 439-449 (2000)
  • Non-Patent Document 12 Drug Discov. Today, 1, 190-198 (1996)
  • An object of the present invention is to provide a novel compound useful as a primer for producing a mucin-type glycopeptide having a borilactosamine skeleton, and a mucin-type glycopeptide having a polyratatosamine skeleton using the primer.
  • the purpose of the present invention is to provide a production method, and to produce mucin-type glycopeptides having a borilactosamine skeleton, which has been difficult to produce so far, which is useful in a wide range of fields such as biochemical research materials, medicines and foods.
  • Another subject of the present invention is a method for producing a glycopeptide having a voralactosamine skeleton by synthesizing a glycopeptide having a desired peptide sequence using a previously synthesized saccharide amino acid having a vorolactosamine skeleton. It is to provide.
  • a novel glycopeptide derivative having an aldehyde group or a ketone group at the end and containing an amino acid residue that can be cleaved by a protease has an aldehyde group or a ketone group.
  • this bond does not degrade under the hydrolysis conditions with proteases, and therefore functions as a primer suitable for the production of mucin-type glycopeptides having a polylactosamine skeleton, And the use of this primer,
  • the present inventors have found that a mucin-type glycopeptide having a ratatosamine skeleton can be easily purified, and that a mucin-type glycopeptide having a borilactosamine skeleton can be produced quickly and in a high yield.
  • the present inventors also protected a sugar amino acid having a pre-synthesized voralactosamine skeleton with a protecting group, and synthesized a sugar peptide having a desired peptide sequence using the protected amino acid.
  • the present inventors have found that a glycopeptide having a borilactosamine skeleton can be produced.
  • the method is independent of proteases or photocleavage. Therefore, this method makes it possible to synthesize glycopeptides containing amino acid residues having properties that are easily cleaved by protease or photocleavage.
  • the present invention also provides a mucin-type glycopeptide that has been useful in a wide range of fields such as biochemical research materials, pharmaceuticals, and foods by the method for producing a glycopeptide using the above-described primer and has been difficult to produce so far.
  • the present invention has been completed.
  • the MUC1 and MUC1 peptide library of the present invention are effective for elucidating the function of MUC1, and the possibility of new drug discovery based on the knowledge obtained therefrom is considered.
  • Studies using glycopeptides include, for example, immobilization of glycopeptide libraries 'chipies', antibody reaction screening, search for specific antibodies, structure-activity relationship investigations in antigen-antibody reactions, specificity's of highly selective monoclonal antibodies Development of antibody drugs and vaccine therapy using glycopeptides is also possible.
  • the present invention provides the following.
  • X is a hydrogen atom, c-c alkyl, c-
  • 6 c represents a reel or chromophore
  • n an integer of 0 to 20;
  • polyacrylamide polymerization degree 1 to: oligo or polypeptide of LO, oxygen atom or NH;
  • A represents an amino acid residue cleavable by a protease
  • A is a sugar amino acid residue substantially free of a site cleavable by a protease, or
  • glycopeptide residue containing any sugar amino acid without a site cleavable by a protease
  • sugar amino acid residue or the glycopeptide residue is represented by the following formula:
  • R 1 and R 6 are each independently hydrogen, N-acetylethylneuraminic acid (Neu5 Ac) group or N-acetylethyldarcosamine (GlcNAc) group;
  • R 2 and R 7 are galactose (Gal) groups
  • R 3 and R 8 are GlcNAc groups
  • R 4 and R 9 are Gal groups
  • R 5 is a GlcNAc group
  • R 10 is an N-acetyl-a-D-galactosamine (GalNAc) group
  • Z 2 and Z 3 are each independently hydrogen or a fucose group
  • n and m are each independently an integer of 0 or more;
  • the bond between R 1 and R 2 is a 2, 3 bond when R 1 is Neu5Ac group, and j81, 3 bond when R 1 is GlcNAc group;
  • R 2 and R 3 are a ⁇ ⁇ , 4 bond
  • R 3 and R 4 are a ⁇ ⁇ , 3 bond
  • R 4 and R 5 are a ⁇ ⁇ , 4 bond
  • R 6 and R 7 are an ⁇ 2,3 bond when R 6 is a Neu5Ac group, and a ⁇ , 3 bond when R 6 is a GlcNA c group;
  • R 7 and R 8 are a ⁇ ⁇ , 4 bond
  • R 8 and R 9 are a ⁇ ⁇ , 3 bond
  • R 9 and R 1G are a ⁇ ⁇ , 3 bond
  • R 5 and R 1G are a ⁇ ⁇ , 6 bond;
  • the bond between Z 1 and R 3 is ⁇ 1, 3 bond;
  • the bond between ⁇ 2 and R 5 is ⁇ 1, 3 bond
  • the bond between ⁇ 3 and R 8 is ⁇ 1, 3 bond
  • the cocoon is a protea derived from Bacillus Licheniformis
  • a compound according to item 1 having an amino acid sequence selected from the group consisting of the amino acid sequences shown.
  • the carrier is:
  • Protected ! which may be a polymer or copolymer of a butyl monomer having an aminooxy group or a hydrazide group, or a polymer having a protected amino acid group or hydrazide group Ethers;
  • Protected ! which may be a silica support having a aminooxy group or a hydrazide group, a resin support, a magnetic bead or a metal support; and
  • R 3 represents a hydroxyl group or an amino group
  • Lys represents a lysine residue
  • Cys represents cysteine
  • n is an integer from 1 to 15 and x: y is 1: 0 to 1: 1000,
  • a N C (— X) — (CH) A -A -A (II)
  • 6 c represents aryl or chromophore
  • n an integer of 0 to 20;
  • polyacrylamide polymerization degree 1 to: oligo or polypeptide of LO, oxygen atom or NH;
  • A is a protease derived from Bacillus Licheniformis.
  • A is a sugar amino acid residue substantially free of a site cleavable by a protease, or
  • glycopeptide residue containing any sugar amino acid without a site cleavable by a protease
  • sugar amino acid residue or the glycopeptide residue is represented by the following formula:
  • R and R are each independently hydrogen, Neu5Ac group or GlcNAc group;
  • R 2 and R 7 are Gal groups
  • R 3 and R 8 are GlcNAc groups
  • R 4 and R 9 are Gal groups
  • R 5 is a GlcNAc group
  • R 10 is a GalNAc group
  • Z 2 and Z 3 are each independently hydrogen or a Fuc group; and n and m are each independently an integer of 0 or more;
  • the bond between R 1 and R 2 is a 2, 3 bond when R 1 is Neu5Ac group, and j81, 3 bond when R 1 is GlcNAc group;
  • R 2 and R 3 are a ⁇ ⁇ , 4 bond
  • R 3 and R 4 are a ⁇ ⁇ , 3 bond
  • R 4 and R 5 are a ⁇ ⁇ , 4 bond
  • R 6 and R 7 are an ⁇ 2,3 bond when R 6 is a Neu5Ac group, and a ⁇ , 3 bond when R 6 is a GlcNA c group;
  • R 7 and R 8 are a ⁇ ⁇ , 4 bond
  • R 8 and R 9 are a ⁇ ⁇ , 3 bond
  • R 9 and R 1G are a ⁇ ⁇ , 3 bond
  • R 5 and R 1G are a ⁇ ⁇ , 6 bond
  • the bond between ⁇ 1 and R 3 is ⁇ 1 , 3 bond;
  • the bond between ⁇ 2 and R 5 is ⁇ 1, 3 bond
  • the bond between ⁇ 3 and R 8 is ⁇ 1, 3 bond
  • A Having a sugar residue represented by: A is the following formula:
  • s is an integer of 1 to 15, and x: y is a group represented by 1: 0 to 1: 1000).
  • (D) A method for producing a glycopeptide, comprising a step of allowing a protease to act on a compound in which a sugar residue is transferred and a sugar chain is extended.
  • (C) A method for producing a glycopeptide, comprising a step of allowing a protease to act on a compound having a sugar residue transferred and a sugar chain extended.
  • Step (B) Step (A) is repeated once or twice or more to extend the sugar chain
  • step (B) The compound obtained in step (A) and an optionally protected aminooxy group, N-alkylaminooxy group, hydrazide capable of reacting specifically with the ketone residue or aldehyde residue of the compound Reacting with a carrier comprising a functional group selected from the group consisting of a group, an azide group, a thiosemicarbazide group, a 1,2-dithiol group and a cysteine residue;
  • Step (B) By reacting the compound obtained in step (B) with a glycosyltransferase in the presence of a sugar nucleotide, a sugar residue is transferred from the sugar nucleotide to the compound, and the sugar chain is elongated.
  • (E) A method for producing a glycopeptide, comprising a step of allowing a protease to act on a compound in which a sugar residue is transferred and a sugar chain is elongated.
  • step (B) The compound obtained in step (A) and an optionally protected aminooxy group, N-alkylaminooxy group, hydrazide group capable of reacting specifically with the ketone residue or aldehyde residue of the compound And a carrier containing a functional group selected from the group consisting of an azide group, a thiosemicarbazide group, a 1,2-dithiol group and a cysteine residue, and at the same time, the step (A) Removing unreacted material in the step;
  • step (C) A sugar transfer enzyme is allowed to act on the compound bound to the carrier obtained in step (B) in the presence of a sugar nucleotide to transfer a sugar residue from the sugar nucleotide to the compound, A step of obtaining a compound in which a sugar chain is extended, the sugar nucleotide having a sugar residue selected from the group consisting of a Gal group, a GlcNAc group, a Fuc group and a Neu5Ac group;
  • Step (D) Step (C) is repeated once or twice or more to extend the sugar chain
  • (F) A method for producing a glycopeptide, comprising the step of causing a protease to act on a compound in which a plurality of sugar residues are transferred and the sugar chain is elongated.
  • X is a hydrogen atom, c1-c alkyl
  • 30 c represents 6 to c aryl or chromophore
  • n an integer of 0 to 20;
  • A represents a linker having a length of 1 to 20 methylene chains
  • Step (B) is repeated one or more times as necessary to extend the sugar chain;
  • a compound in which the sugar residue is transferred and the sugar chain is extended, and the ketone residue of the compound Protected which can react specifically with a group or aldehyde residue, may be an aminooxy group, N-alkylaminooxy group, hydrazide group, azide group, thiosemicarbazide group, 1,2-dithiol group and cysteine. Reacting with a carrier containing a functional group selected from the group consisting of residues ;and
  • a process for producing a glycopeptide comprising
  • Step (B) Step (A) is repeated one or more times as necessary to extend the sugar chain as necessary;
  • (C) a compound in which a sugar residue is transferred and a sugar chain is extended, and a protected compound capable of reacting specifically with a ketone residue or an aldehyde residue of the compound, and may be an aminooxy group, N-alkyl Reacting a carrier containing a functional group selected from the group consisting of an aminooxy group, a hydrazide group, an azide group, a thiosemicarbazide group, a 1,2-dithiol group and a cysteine residue;
  • (E) A method for producing a glycopeptide, comprising a step of allowing a protease to act on a compound in which a sugar residue is transferred and a sugar chain is elongated.
  • Said glycopeptide has the following formula:
  • each X 1 independently represents a hydrogen atom or the following formula:
  • R 1 and R 6 are each independently hydrogen, Neu5Ac group or GlcNAc group;
  • R 2 and R 7 are Gal groups
  • R 3 and R 8 are GlcNAc groups
  • R 4 and R 9 are Gal groups
  • R 5 is a GlcNAc group
  • R 10 is a GalNAc group
  • Z 2 and Z 3 are each independently hydrogen or a Fuc group; and n and m are each independently an integer of 0 or more;
  • the bond between R 1 and R 2 is a 2, 3 bond when R 1 is Neu5Ac group, and j81, 3 bond when R 1 is GlcNAc group;
  • R 2 and R 3 are a ⁇ ⁇ , 4 bond
  • R 3 and R 4 are a ⁇ ⁇ , 3 bond
  • R 4 and R 5 are a ⁇ ⁇ , 4 bond
  • R 6 and R 7 are an ⁇ 2,3 bond when R 6 is a Neu5Ac group, and a ⁇ , 3 bond when R 6 is a GlcNA c group;
  • R 7 and R 8 are a ⁇ ⁇ , 4 bond
  • R 8 and R 9 are a ⁇ ⁇ , 3 bond;
  • the bond between R 9 and R 1G is a ⁇ ,, 3 bond;
  • R 5 and R 1G are a ⁇ ,, 6 bond
  • the bond between ⁇ 1 and R 3 is ⁇ 1 , 3 bond;
  • the bond between ⁇ 2 and R 5 is ⁇ 1, 3 bond
  • the bond between ⁇ 3 and R 8 is ⁇ 1, 3 bond
  • ⁇ 1 represents a hydrogen atom, acetyl, acyl, alkyl or aryl
  • ⁇ 2 represents a hydroxyl group, ⁇ , alkyl or aryl.
  • each X 1 independently represents a hydrogen atom or the following formula:
  • R 1 and R 6 are each independently hydrogen, Neu5 Ac group or GlcNAc group. Yes;
  • R 2 and R 7 are Gal groups
  • R 3 and R 8 are GlcNAc groups
  • R 4 and R 9 are Gal groups
  • R 5 is a GlcNAc group
  • R 10 is a GalNAc group
  • Z 2 and Z 3 are each independently hydrogen or a Fuc group; and n and m are each independently an integer of 0 or more;
  • the bond between R 1 and R 2 is a 2, 3 bond when R 1 is Neu5Ac group, and j81, 3 bond when R 1 is GlcNAc group;
  • R 2 and R 3 are a ⁇ ⁇ , 4 bond
  • R 3 and R 4 are a ⁇ ⁇ , 3 bond
  • R 4 and R 5 are a ⁇ ⁇ , 4 bond
  • R 6 and R 7 are an ⁇ 2,3 bond when R 6 is a Neu5Ac group, and a ⁇ , 3 bond when R 6 is a GlcNA c group;
  • R 7 and R 8 are a ⁇ ⁇ , 4 bond
  • R 8 and R 9 are a ⁇ ⁇ , 3 bond
  • R 9 and R 10 are a ⁇ ⁇ , 3 bond
  • R 5 and R 1G are a ⁇ ⁇ , 6 bond
  • the bond between ⁇ 1 and R 3 is ⁇ 1 , 3 bond;
  • the bond between ⁇ 2 and R 5 is ⁇ 1, 3 bond
  • the bond between ⁇ 3 and R 8 is ⁇ 1, 3 bond
  • ⁇ 1 represents a hydrogen atom, acetyl, acyl, alkyl or aryl
  • ⁇ 2 represents a hydroxyl group, ⁇ , alkyl or aryl.
  • a method for producing a desired glycopeptide comprising the following steps:
  • R 1 and R 6 are each independently hydrogen, Neu5Ac group or GlcNAc group;
  • R 2 and R 7 are Gal groups
  • R 3 and R 8 are GlcNAc groups
  • R 4 and R 9 are Gal groups
  • R 5 is a GlcNAc group
  • R 10 is a GalNAc group
  • R 11 is a hydrogen atom or a methyl group
  • Z 2 and Z 3 are each independently hydrogen or a Fuc group; and n and m are each independently an integer of 0 or more;
  • the bond between R 1 and R 2 is a 2, 3 bond when R 1 is Neu5Ac group, and j8 1, 3 bond when R 1 is GlcNAc group;
  • R 2 and R 3 are a ⁇ ,, 4 bond
  • R 3 and R 4 are a ⁇ ,, 3 bond
  • R 4 and R 5 are a ⁇ ,, 4 bond
  • R 6 and R 7 are an ⁇ 2,3 bond when R 6 is a Neu5Ac group, and a,, 3 bond when R 6 is a GlcNA c group;
  • the bond between R 7 and R 8 is a ⁇ ,, 4 bond;
  • Bond between R 8 and R 9, ⁇ ⁇ , is 3 bonds;
  • R 9 and R 1G are a ⁇ ,, 3 bond
  • R 5 and R 1G are a ⁇ ,, 6 bond
  • the bond between ⁇ 1 and R 3 is ⁇ 1 , 3 bond;
  • the bond between ⁇ 2 and R 5 is ⁇ 1, 3 bond
  • the bond between ⁇ 3 and R 8 is ⁇ 1, 3 bond
  • ⁇ is a 9-fluormethylcarboxyl group or a tert-butoxycarbol group
  • a sugar amino acid having a sugar chain protected by protecting the sugar chain of the sugar amino acid represented by the following: a acetyl group, a benzoyl group, a methyl group, a methoxymethyl group, A step selected from the group consisting of trimethylsilyl group, t-butyldimethylsilyl group, dimethylphenol group and triisopropylpropyl group, benzyl group, benzylidene group, isopropylidene group, di-tert-butylsilylidene group power;
  • step (B) Using the sugar amino acid in which the sugar chain obtained in step (A) is protected and an amino acid N-protected with a 9-fluorenylmethyloxyl group or a tert-butoxycarbol group, the desired peptide sequence is prepared. Synthesizing a glycopeptide having a protected sugar chain;
  • step (C) a step of deprotecting the glycopeptide protected in sugar chain obtained in step (B) to produce the desired glycopeptide
  • a sugar chain amino acid containing about 1 to 3 sugars which is relatively easy to prepare, is used for synthesizing a mucin-type glycopeptide having a borilactosamine skeleton, and the sugar chain is elongated after the peptide synthesis. It is possible to synthesize a glycopeptide having a chain and to prepare a library of each sugar chain structure that is an intermediate of a sugar chain elongation reaction. Furthermore, since the sugar chain elongation reaction is carried out by carrying a glycopeptide on a water-soluble polymer, the acceleration effect of the reaction and the simplification of the molecular operation become possible, and the sugar chain elongation reaction can be automated.
  • the present invention makes it possible to synthesize mucin-type glycopeptides that are useful in a wide range of fields such as biochemical research materials, pharmaceuticals, and foods, and that have been difficult to produce.
  • the present invention also protects a sugar amino acid having a pre-synthesized voralactosamine skeleton with a protecting group, and synthesizes a glycopeptide having a desired peptide sequence using the protected amino acid, thereby It enables the production of glycopeptides having The method is independent of proteases or photocleavage. As a result, it is possible to synthesize a glycopeptide containing an amino acid residue having the property of being easily cleaved by protease or photocleavage.
  • glycopeptide library can be used as a standard sample for structural analysis and biochemical tests.
  • this glycopeptide library can be placed on a chip for comprehensive detection of glycopeptide recognition proteins, pathological diagnosis, cell adhesion sequence search, sequence analysis related to cell growth and apoptosis, etc. become.
  • FIG. 1 shows the procedure up to the production of the glycopeptide of the present invention.
  • the indicated numbers indicate the compound numbers in the examples.
  • SEQ ID NO: 1 to 20 Partial amino acid sequence of mucin type glycoprotein MUC1 10 residues
  • SEQ ID NO: 21 to 40 Partial amino acid sequence of mucin type glycoprotein MUC1
  • SEQ ID NO: 41 to 60 Mucin type glycoprotein MUC 1
  • a partial amino acid sequence of 20 residues of SEQ ID NOs: 61 to 66 Examples of amino acid sequences contained in a carrier contained in a compound BEST MODE FOR CARRYING OUT THE INVENTION
  • sucgar amino acid means a combination of a sugar residue and an amino acid residue, and is used interchangeably with “sugar amino acid residue”.
  • sugar amino acid residue substantially not including a site cleavable by a protease means that a compound represented by the above item (4) is treated with a protease. Even so, it refers to a sugar amino acid residue in which the sugar amino acid moiety is not cleaved by more than 50% by protease, preferably a sucrose amino acid residue not cleaved by more than 20%.
  • glycopeptide residue means a peptide residue containing at least one sugar amino acid, and is used interchangeably with “glycopeptide”.
  • the sugar residue constituting the sugar amino acid contained in the glycopeptide residue is not particularly limited, but monosaccharide to monosaccharide or monosaccharide to trisaccharide derivatives are preferred. Inductive materials are more preferably used.
  • sugar chain refers to a compound comprising one or more unit sugars (monosaccharide and Z or a derivative thereof). When two or more unit sugars are connected, each unit sugar is linked by dehydration condensation using a glycosidic bond.
  • unit sugars monosaccharide and Z or a derivative thereof.
  • sugar chains include polysaccharides contained in the living body (glucose, galactose, mannose, fucose, etc.
  • sugar chain may include both sugar chains and sugar chain-containing substances.
  • monosaccharide refers to a polyhydroxy aldehyde or polyhydroxy ketone and a hydrolyzate thereof that is not hydrolyzed into a simpler molecule and contains at least one hydroxyl group and at least one aldehyde group or ketone group. Refers to a derivative.
  • monosaccharides are represented by the general formula C H O, but are not limited to them: fucose (deoxyhexose), N n n 2n n
  • cetinoregenorecosamine cetinoregenorecosamine.
  • Otatos nonose and decourse. It is generally equivalent to an aldehyde or ketone of a chain polyhydric alcohol. The former is called aldose and the latter is called ketose.
  • galactose refers to any isomer, but is typically j8-D galactose, and is used to refer to j8-D-galactose unless otherwise specified.
  • acetylyldarcosamine refers to any isomer, but is typically N-acetyleno ⁇ D darcosamine, and unless otherwise specified, ⁇ acetylthio ⁇ -D— Used to refer to darcosamine.
  • fucose refers to any isomer, but is typically a L-fucose, and is used to refer to ⁇ L-fucose unless otherwise specified.
  • sialic acid is a general term for sialic acid and derivatives of neuraminic acid.
  • N-acyl (N-acetyl or N-glycolyl) neuraminic acid and N-acyl-0-acetylethylneuraminic acid are naturally known.
  • sialic acid is rarely present in the free state, it is mostly an acid-labile bond ( ⁇ -ketoside bond) in a monosaccharide, polysaccharide, glycoprotein, glycopeptide, or glycosphingolipid molecule.
  • the sialic acid used herein is preferably acetylethylneuraminic acid.
  • acetylylgalatatosamine refers to any isomer, but is typically ⁇ acetylyl-a-D galactosamine, and unless otherwise specified, N-acetylyl-a-D galactosamine. Used as a pointer.
  • sugar symbols, designations, abbreviations (Glc and the like) and the like are different when representing a monosaccharide and when used in a sugar chain. In the sugar chain, the unit sugar has a dehydration condensation with another unit sugar to which it is bonded, so that the mutual force also exists in a form excluding hydrogen or hydroxyl group.
  • a “sugar residue” for a sugar refers to those with 1 to n hydrogens, usually because sugars are linked to other chemical moieties via hydroxyl groups.
  • a sugar residue is a hydroxyl group of a sugar that has a hydrogen power of ⁇ to n (in this case, n is the number of hydroxyl groups present in the sugar), but is not limited to this. If there is, it can indicate that the hydrogen has been removed.
  • sucgar residue may be a monovalent, divalent, or trivalent to nvalent residue depending on the situation in which it is used.
  • specific names are expressed in the form of “sugar name” + “group”, for example, galactose group, Gal group, and the like.
  • Monosaccharides are generally joined by glycosidic bonds to form disaccharides and polysaccharides.
  • the direction of the bond with respect to the plane of the ring is indicated by ⁇ and j8.
  • Also described are specific carbon atoms that form a bond between two carbons.
  • Branches of sugar chains are represented by parentheses, and are arranged and placed immediately to the right of the unit sugar to be bound. For example,
  • C-6 of N-acetylyldarcosamine is bonded to C-1 of N-acetylyldarcosamine with a j8 glycoside
  • C-3 of N-acetylyldarcosamine is further linked to C-1 and 13 glycosides of galactose.
  • GlcNAc (3-1 ⁇ Gal) 6-1 ⁇ 8 GlcNAc.
  • Monosaccharides are represented on the basis of numbering as small as possible (latent) carbonyl groups. According to the general standard of organic chemical nomenclature, even when an atomic group superior to a (latent) carbon group is introduced into a molecule, it is usually represented by the above numbering.
  • a “monosaccharide derivative” refers to a substance that results from the substitution of one or more hydroxyl groups on an unsubstituted monosaccharide with another substituent. That Examples of such monosaccharide derivatives include saccharides having a carboxyl group (for example, aldonic acids in which the C-1 position is oxidized to form carboxylic acids (for example, D-dalconic acid in which D-glucose is oxidized), terminal C Uronic acid (D-glucose oxidized D-glucuronic acid) with carboxylic acid atom, sugar having amino group or amino group derivative (eg acetylated amino group) (eg N-acetyl- D-darcosamine, N-acetyl-D-galatatosamine, etc., sugars that have both amino and carboxyl groups (eg, Neu5Ac (sialic acid), N-acetylethylmuramic acid, etc.),
  • the “amino acid residue” constituting the glycopeptide residue of the present invention is not particularly limited as long as it has an amino group and a carboxy group in the molecule.
  • the Gly (glycine) residue and the Ala (alanine) residue are not particularly limited.
  • Val (parin) residue Leu (leucine) residue, lie (isoleucine) residue, Tyr (tyrosine) residue, Trp (tributophane) residue, Glu (glutamic acid) residue, Asp (aspartic acid) residue Group, Lys (lysine) residue, Arg (arginine) residue, His (histidine) residue, Cys (cystine) residue, Met (methionine) residue, Ser (serine) residue, Thr (threonine) residue And a-amino acid residues such as Asn (asnolagin) residue, Gin (glutamine) residue or Pro (proline) residue, or
  • 8-amino acid residue such as j8-Ala residue, etc.
  • the amino acid residue may be either D-form or L-form, but the L-form is preferred.
  • the glycopeptide residue the above-mentioned amino acid residues or 2-30 glycopeptide residues are preferred. More preferred are glycopeptide residues having 4 to 20 forces.
  • Gly or the like may be omitted when it is clear that a residue is indicated.
  • amino acid is a peptide bond
  • structure is as follows:
  • R represents a side chain. For example, if both R are H,
  • Gly-Gly or “-Gly” located at the end of the peptide chain represents a monovalent glycine residue.
  • the peptide chain When the constituent amino acid residue includes a proline residue and is expressed as Gly—Pro, the structure is as follows:
  • R A to R e represent side chains.
  • R A is H
  • R B force SCH
  • Re is CH (CH) CH
  • the peptide chain is also represented as Gly—Ala—Val.
  • Gly— represents a monovalent glycine residue
  • —Ala— represents a divalent alanine residue
  • —Val represents a monovalent palin residue
  • the serine residue or threonine residue is "Ser-” or " It is expressed as “Ser”, and the threonine residue is expressed as “Thr-” or “-Thr” and is a monovalent amino acid residue.
  • Sugar chains may be bound to these amino acid residues. When sugar chains are attached, these residues are divalent, and serine and threonine residues are as follows:
  • sugar amino acid of the present invention is not particularly limited as long as the amino acid residues and sugar residues listed above can theoretically be combined, but preferably, the combination is not limited. ,
  • n an integer of 1 to 10
  • Gal represents galactose
  • Glc represents gnoleose
  • Man represents mannose
  • Xyl represents xylose
  • GlcNAc represents N-acetyl D darcosamine
  • GalNAc represents N-acetyl D represents galactosamine.
  • N-terminal means an amino group that may be substituted at the end of the peptide main chain.
  • C-terminus refers to a substitution located at the end of the peptide backbone! And, it means a carboxyl group.
  • the “side chain” means a functional group extending from the peptide main chain in a direction orthogonal to the direction in which the peptide main chain extends or a portion containing the functional group.
  • the “primer” means a substance having an action that triggers the initiation of a reaction in an enzyme reaction.
  • transferase is a general term for enzymes that catalyze a group transfer reaction.
  • transferase may be used interchangeably with “transferase”.
  • the group transfer reaction is represented by the following formula (1): X-Y + ZH XH + ZY (1)
  • the group Y is transferred from one compound (donor) to another compound (acceptor).
  • glycosyltransferase refers to a sugar (corresponding to group Y in formula (1) above; unit sugar or sugar chain) separated from a certain place (corresponding to compound X—Y in formula (1) above).
  • the enzyme has an action of catalyzing the transfer to the site (corresponding to the compound Z—H of the above formula (1)).
  • examples of the glycosyltransferase include galactose transferase, glucose transferase, sialic acid transferase, mannose transferase, fucose transferase, xylose transferase, N-acetylylcosamine transferase, and N-acetyl galatatosamine transferase. However, it is not limited to them.
  • glycosyltransferases are specific for a particular binding mode.
  • j81,3-N-acetylylcosamine transferase is a glycosyltransferase that binds the 3rd position of a sugar such as galactose and the 1st position of N-acetylethyldarcosamine.
  • the glycosyltransferase used in the present invention is only required to be able to use sugar nucleotides as a sugar donor.
  • sugar chain elongation reaction refers to a reaction in which the chain length of a sugar chain is elongated in the presence of a glycosyltransferase as defined above.
  • biomolecule refers to a molecule related to a living body.
  • biomolecules are sometimes referred to herein as biological samples.
  • living body refers to a biological organism, including but not limited to animals, plants, fungi, viruses, and the like. Therefore, a biomolecule includes a molecule extracted from a living body, but is not limited thereto, and any molecule that can affect a living body falls within the definition of a biomolecule.
  • biomolecules include proteins, polypeptides, oligopeptides, peptides, glycopeptides, polynucleotides, oligonucleotides, nucleotides, sugar nucleotides, nucleic acids (eg, DNA such as cDNA, genomic DNA, mRNA Such as RNA), polysaccharides, oligosaccharides, lipids, small molecules (eg, hormones, ligands, signaling substances, small organic molecules, etc.), complex molecules thereof, and the like.
  • the biomolecule may preferably be a sugar chain or a complex molecule containing a sugar chain (eg, glycoprotein, glycolipid, etc.).
  • the source of such biomolecules may be any animal, plant, bacteria, or virus, as long as it is a material to which a biological sugar chain is bound or attached. More preferably, an animal-derived biological sample is used. Preferable examples include whole blood, plasma, serum, sweat, saliva, urine, knee fluid, amniotic fluid, and cerebrospinal fluid, and more preferable examples include plasma, serum, and urine.
  • Biological samples include biological samples that have not been previously separated from individuals. For example, it includes mucosal tissue that can be contacted with a test solution from the outside, or glandular tissue, preferably ductal epithelium attached to breast, prostate, and spleen.
  • the terms "protein”, “polypeptide”, “oligopeptide” and “peptide” are used interchangeably herein and refer to a polymer of amino acids of any length. .
  • the polymer may be linear or branched or cyclic.
  • the amino acid may be a modified amino acid, which may be natural or non-natural.
  • the term also includes assembly into a complex of multiple polypeptide chains. May be included.
  • the term also encompasses natural or artificially modified amino acid polymers. Such modifications include, for example, disulfide bond formation, daricosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification (eg, conjugation with a labeling component).
  • This definition also includes, for example, polypeptides containing one or more analogs of amino acids (eg, including non-natural amino acids, etc.), peptidomimetic compounds (eg, peptoids), and the art! Other modifications are included
  • sugar nucleotide means a nucleotide to which a sugar residue as defined above is bound, and the sugar nucleotide used in the present invention can be used by the above enzyme. If it is, it will not specifically limit.
  • uridine 5 monodiphosphate galactose, uridine 5, monodiphosphate-N acetyl dalcosamine, uridine 5, mono diphosphate N acetyl galactosamine, uridine-5, monodiphosphate glucuronic acid, uridine 5, 1 Examples include diphosphate xylose, guanosine 5, monodiphosphate fucose, guanosine 5, monodiphosphate mannose, cytidine-5, monomonophosphate 1N acetylneuraminic acid, and sodium salts thereof.
  • substitution refers to replacement of one or more hydrogen atoms in an organic compound or substituent with another atom or atomic group.
  • One hydrogen atom can be removed and substituted with a monovalent substituent, and two hydrogen atoms can be removed and substituted with a divalent substituent.
  • alkyl refers to a monovalent group formed by loss of one hydrogen atom in an aliphatic hydrocarbon (alkane) force such as methane, ethane, or propane.
  • n 2n + l is represented by one (where n is a positive integer).
  • Alkyl can be linear or branched.
  • substituted alkyl refers to an alkyl in which one or more hydrogen atoms are each independently substituted with a substituent as defined below.
  • C1-C2 alkyl C1-C3 alkyl, C1-C4 alkyl, C1-C5 alkyl, C1-C6 alkyl, C1-C7 alkyl, C1-C8 alkyl, C1-C9 alkyl, C1-C10 alkyl, C1-C11 alkyl C1-C12 alkyl, C1-C15 alkyl, Cl-C20 alkyl, C1-C25 alkyl or C1-C30 alkyl.
  • C 1 -C 10 alkyl means a linear or branched alkyl having 1 to 10 carbon atoms, such as methyl (CH—), ethyl (CH 1), n-propyl. (CH CH CH one)
  • aryl refers to a monovalent aromatic of 6 to 30 carbon atoms derived by removing one hydrogen atom from one carbon atom of the parent aromatic ring system. This refers to the hydrocarbon radical. Representative aryl groups include, but are not limited to, benzene, naphthalene, anthracene, biphenyl and the like.
  • chromophore refers to a functional group having an absorption band in the ultraviolet light or visible light region, or emitted light in the visible light region excited by electromagnetic waves in the ultraviolet light or visible light region.
  • a functional group that emits For example, nitro group, benzyl group, thiophenol group, paranitrophenol group, 2,4 di-trifluoro group, dansyl group, 2-aminobenzyl group, fluorescein isothiocyanate (FITC) group, 4-methoxy group Examples thereof include, but are not limited to, ⁇ -naphthylamide group.
  • keto acid is a general term for compounds having a carboxyl group and a carbocycle group of a ketone.
  • aldehyde acid is a general term for compounds having a carboxyl group and a carboxylic group of an aldehyde.
  • 1 30 6 30 N represents an integer of 0 to 20;
  • A represents a linker having a length of 1 to 20 methylene chains).
  • protecting reaction refers to a reaction in which a protective group such as Boc (tert-butoxycarbonyl group) is added to a functional group desired to be protected.
  • a protective group such as Boc (tert-butoxycarbonyl group)
  • Boc tert-butoxycarbonyl group
  • deprotection reaction refers to a reaction for removing a protecting group such as Boc.
  • Examples of deprotection reactions include reactions with trifluoroacetic acid (TFA) and reduction reactions with PdZC.
  • examples of the “protecting group” include, for example, a fluorenylmethoxycarbol (F moc) group, a acetyl group, a benzyl group, a benzoyl group, a tert-butoxycarbol group, t-Butyldimethyl group, silyl group, trimethylsilylethyl group, N-phthalimidyl group, trimethylsilylethyloxycarbonyl group, 2-trow 4,5 dimethoxybenzyl group, 2-trow 4,5-dimethoxybenzyloxy Carbon group, strong rubamate group, methyl group, methoxymethyl group, trimethylsilyl group, t-butyldimethylsilyl group, dimethylphenyl group, triisopropylpropylsilyl group, benzylidene group, isopropylidene group, ditert-butylsilylidene group, etc.
  • F moc fluorenylmethoxycar
  • the protecting group can be used, for example, to protect a reactive functional group such as an amino group or a force lpoxyl group. Depending on the reaction conditions and purpose, various protecting groups can be used properly.
  • a trimethylsilylethyloxycarboxyl group, a 2-trow 4,5 dimethoxybenzyloxycarboxyl group or a derivative thereof is preferable.
  • the target product is a contaminant (unreacted weight loss, by-product, solvent, etc.) from the reaction solution, and a method commonly used in the art (for example, extraction, distillation, After removal by washing, concentration, precipitation, filtration, drying, etc., followed by treatment by a combination of post-treatment methods commonly used in the art (eg adsorption, elution, distillation, precipitation, precipitation, chromatography, etc.) obtain.
  • a method commonly used in the art for example, extraction, distillation, After removal by washing, concentration, precipitation, filtration, drying, etc., followed by treatment by a combination of post-treatment methods commonly used in the art (eg adsorption, elution, distillation, precipitation, precipitation, chromatography, etc.) obtain.
  • the present invention provides the following formula:
  • X represents a hydrogen atom, C-C alkyl, C-C aryl or chromophore
  • A represents a sugar amino acid substantially free of a site cleavable by a protease.
  • glycopeptide residue containing any sugar amino acid without a site cleavable by a protease By using this as a primer, purification of a glycopeptide, which conventionally required multi-step purification, can be simplified, and the glycopeptide can be produced quickly and with high yield. Since the compound of the above formula (I) of the present invention always has an aldehyde group or a ketone group at the terminal, it may be protected with an aminooxy group, an N alkylaminooxy group, a hydrazide group, an azide group, a thiosemicarbazide group.
  • 1,2-dithiol group and a carrier containing a functional group selected from the group consisting of cysteine residues by reacting with a carrier containing a functional group selected from the group consisting of the above-mentioned formula (I) and using it as a polymer primer can do.
  • the bond obtained by this reaction is a strong bond that does not decompose under the subsequent hydrolysis with a protease (pH condition, etc.), so that there is an advantage that the purification of hydrolysis is very simple.
  • the combination with the cleavable amino acid residue (A 2 ) is a bond produced by the reaction of at least the terminal aldehyde or ketone group of the compound of formula (I) with the carrier in a pH range where hydrolysis by a protease can occur. Any combination is acceptable as long as it does not decompose. Amino acid residues force of some or all and A 2 of the polypeptide of A 1 also Narupepu tide recognizing protease may also be used.
  • Such combinations include a combination of a protease (glutamidase) derived from Bacillus Licheniformis and a glutamic acid residue or cysteine residue that can be cleaved by this protease; Combination of peptidase and Asn (recognition site (A 2 )) (cleaves the C-terminus of asparagine (Asn)); Combination of arginyl endopeptidase and Arg (recognition site (A 2 )) (arginine ( Cleaves C-terminal of Arg)); Combination of Achromopacter protease I and lysine (Lys) (recognition site (A 2 )) (cleaves C-terminal of lysine (Lys)); , Arginine (Arg) or lysine (Lys) (recognition site (A 2 )) combination (when Arg is recognized, C-terminal of arginine (Arg) is a
  • Trp When Trp is recognized, the C-terminal end of tributophan (Trp) is cleaved.); V8 protease and Glu (recognition site (A 2 )) (Cleaves the C-terminus of glutamic acid (Glu)); Factor Xa (factor Xa) and —lie—Glu—Gly—Arg— (recognition site, according to the definition in this document, the recognition site ( A 2 ) is arginine (Arg) and —lie—Glu—G1 y— is the end of A 1 ; this cleaves the C terminus of arginine (Arg).) Enterokinase and Asp— Asp— Asp— Asp— Lys— (recognition site, according to the definition of this specification, the recognition site (A 2 ) is lysine (Ly a s), -Asp- Asp- A sp- Asp- is the terminus of A 1;.
  • Bacillus spp. In particular by Bacillus lichen niformis ATCC 14580.
  • This strain can be obtained from the American Type Culture Collection (ATCC).
  • ATCC American Type Culture Collection
  • the genomic DNA of Bacillus liquor-formis ATCC 14580 strain can be prepared from cultured cells of the strain according to known methods (M. Stahl et al., Journal of Bacteriology, 154, 406-412 (1983)).
  • HGVTSAPDTR (SEQ ID NO: 2)
  • VTSAPDTRPA SEQ ID NO: 4
  • TRPAPGSTAP SEQ ID NO: 10
  • PAPGSTAPPA SEQ ID NO: 12
  • APGSTAPPAH (SEQ ID NO: 13),
  • HGVTSAPDTRP (SEQ ID NO: 22),
  • GVTSAPDTRPA SEQ ID NO: 23
  • VTSAPDTRPAP (SEQ ID NO: 24),
  • TSAPDTRPAPG (SEQ ID NO: 25),
  • APDTRPAPGST (SEQ ID NO: 27),
  • TRPAPGSTAPP SEQ ID NO: 30
  • PAPGSTAPPAH (SEQ ID NO: 32),
  • PAHGVTSAPDT (SEQ ID NO: 40),
  • HGVTSAPDTRPAPGSTAPPA (SEQ ID NO: 41), GVTSAPDTRPAPGSTAPPAH (SEQ ID NO: 42), VTSAPDTRPAPGSTAPPAHG (SEQ ID NO: 43), TSAPDTRPAPGSTAPPAHGV (SEQ ID NO: 44), SAPDTRPAPGSTAPPAHGVT (SEQ ID NO: 45), APDTRPAPGSTAPPAHGVTS (SEQ ID NO: 46), PDPA, TRPA No. DTRPAPGSTAPPAHGVTSAP (SEQ ID NO: 48),
  • TRPAPGSTAPPAHGVTSAPD (SEQ ID NO: 49),
  • RPAPGSTAPPAHGVTS APDT (SEQ ID NO: 50)
  • PAPGSTAPPAHGVTSAPDTR SEQ ID NO: 51
  • APGSTAPPAHGVTSAPDTRP (SEQ ID NO: 52),
  • PAHGVTSAPDTRPAPGSTAP (SEQ ID NO: 59)
  • AHGVTSAPDTRPAPGSTAPP (SEQ ID NO: 60),
  • the polymer carrier that can be used in the present invention can bind the group represented by the formula (I), and after the coupling, the compound of formula (I) can act by the action of glycosyltransferase as described below.
  • a vinyl monomer having an aminooxy group or a hydrazide group may be protected.
  • Polymers or copolymers of the above include acrylamides, methacrylate amides, acrylic acids, methacrylic acids, styrenes, fatty acid butyl esters, etc.
  • Polyethers which may have an aminooxy group or hydrazide group; silica carriers, rosin carriers, magnetic beads or amino beads having an aminooxy group or hydrazide group which may be protected
  • Metallic substrate e.g., the following expression:
  • the above-mentioned polymer or copolymer of a bull monomer having an aminooxy group or a hydrazide group which may be protected is at least one of a polymer or copolymer of an unsubstituted vinyl monomer.
  • acrylamides examples include N-alkyl acrylamides such as acrylamide, N-ethyl acrylamide, N-isopropyl acrylamide, etc., which may have protected amino group or hydrazide group. Illustrated.
  • methacrylamides examples include methacrylamide, N-methyl methacrylamide N-ethyl methacrylamide, N-isopropyl methacrylamide, etc., which may have an aminooxy group or a hydrazide group which may be protected.
  • N-alkyl methacrylamide and the like are exemplified.
  • acrylic acid examples include acrylic acid such as acrylic acid, methyl acrylate, ethyl acrylate, hydroxyethyl acrylate, and dimethylaminoethyl acrylate, which may have an aminooxy group or hydrazide group which may be protected.
  • acrylic acid such as acrylic acid, methyl acrylate, ethyl acrylate, hydroxyethyl acrylate, and dimethylaminoethyl acrylate, which may have an aminooxy group or hydrazide group which may be protected.
  • acid esters examples include acid esters.
  • the above methacrylic acids are protected and may have an aminooxy group or a hydrazide group.
  • Examples include methacrylic acid esters.
  • styrenes examples include styrene, p-hydroxystyrene, p-hydroxymethylstyrene and the like, which may be protected but may have an aminooxy group or a hydrazide group.
  • fatty acid vinyl ester examples include vinyl acetate and vinyl butyrate which may be protected and have an aminooxy group or a hydrazide group.
  • the polymer or copolymer of fatty acid bule ester in the present invention includes an alkali after polymerization reaction, etc. In this case, all or part of the ester bond is hydrolyzed.
  • polyethers may be protected! Polyethylene glycol which may have an aminooxy group or a hydrazide group, or may be protected, and may have an aminooxy group or a hydrazide group. Examples thereof include polyethylene and polyethylene glycol substituted with aryl groups.
  • the polymer carrier here may be either water-insoluble or water-soluble, but water-soluble is preferred.
  • the general molecular weight is about 10,000 to about 5000000, preferably 20000 to 2000000, more preferably ⁇ 50,000 to 1000000.
  • the form includes a bead shape, a fiber shape, a film shape, and a film shape, but is not particularly limited.
  • n is an integer of 1 to 15, preferably 1 to 10, and more preferably 1 to 5.
  • the ratio of x: y is 1: 0 to 1: 1000, preferably 1: 0 to 1: 100.
  • the molecular weight of the polymer carrier is about 10,000 to about 5000000, preferably ⁇ is 20000 to 2000000, more preferably ⁇ is 50000 to 1000000.
  • X represents a hydrogen atom, c-c alkyl, c-c aryl or chromophore
  • n an integer of 0 to 20;
  • polyacrylamide polymerization degree 1 to: oligo or polypeptide of LO, oxygen atom or NH;
  • A represents an amino acid residue cleavable by a protease
  • A is a sugar amino acid residue substantially free of a site cleavable by a protease, or
  • glycopeptide residue containing any sugar amino acid without a site cleavable by a protease
  • sugar amino acid residue or the glycopeptide residue is represented by the following formula:
  • R 1 and R 6 are each independently hydrogen, Neu5Ac group or GlcNAc group;
  • R 2 and R 7 are Gal groups
  • R 3 and R 8 are GlcNAc groups
  • R 4 and R 9 are Gal groups
  • R 5 is a GlcNAc group
  • R 10 is a GalNAc group
  • ZZ 2 and Z 3 are each independently hydrogen or a Fuc group; and n and m are each independently an integer greater than or equal to 0;
  • the bond between R 1 and R 2 is a 2, 3 bond when R 1 is Neu5Ac group, and j81, 3 bond when R 1 is GlcNAc group;
  • R 2 and R 3 are a ⁇ ⁇ , 4 bond
  • R 3 and R 4 is a ⁇ ⁇ , 3 bond
  • the bond between R 4 and R 5 is a ⁇ ⁇ , 4 bond
  • R 6 and R 7 are an ⁇ 2,3 bond when R 6 is a Neu5Ac group, and a ⁇ ⁇ , 3 bond when R 6 is a GlcNA c group;
  • R 7 and R 8 are a ⁇ ⁇ , 4 bond
  • R 8 and R 9 are a ⁇ ⁇ , 3 bond
  • R 9 and R 1G are a ⁇ ⁇ , 3 bond
  • R 5 and R 1G are a ⁇ ⁇ , 6 bond
  • the bond between ⁇ 1 and R 3 is ⁇ 1 , 3 bond;
  • the bond between ⁇ 2 and R 5 is ⁇ 1, 3 bond
  • the bond between ⁇ 3 and R 8 is ⁇ 1, 3 bond
  • the invention provides a compound of the following formula:
  • 30 c 6 c represents a reel or chromophore
  • n an integer of 0 to 20;
  • polyacrylamide polymerization degree 1 to: oligo or polypeptide of LO, oxygen atom or NH;
  • A represents an amino acid residue cleavable by a protease
  • A is a sugar amino acid residue substantially free of a site cleavable by a protease, or
  • glycopeptide residue containing any sugar amino acid without a site cleavable by a protease
  • sugar amino acid residue or the glycopeptide residue is represented by the following formula:
  • R 1 is hydrogen, a sialic acid group or a GlcNAc group
  • R 2 is a Gal group
  • R 3 is a GlcNAc group
  • R 4 is a Gal group
  • R 5 is a GlcNAc group
  • R 10 is a GalNAc group
  • R 8 is hydrogen or a sialic acid group
  • R 9 is a Gal group
  • Z 1 and Z 2 are each independently hydrogen or a Fuc group
  • n is an integer greater than or equal to 0;
  • R 1 and R 2 are an a 2, 3 bond when R 1 is a sialic acid group, and a j8 1, 3 bond when R 1 is a GlcNAc group;
  • R 2 and R 3 are a ⁇ ,, 4 bond
  • R 3 and R 4 are a ⁇ ,, 3 bond
  • R 4 and R 5 are a ⁇ ,, 4 bond
  • R 8 and R 9 are ⁇ 2, 3 bond;
  • R 9 and R 10 are a ⁇ ,, 3 bond
  • R 5 and R 1G are a ⁇ ,, 6 bond
  • the bond between ⁇ 1 and R 3 is ⁇ 1 , 3 bond
  • the present invention provides the following formula:
  • ⁇ N C (— X) — (CH) A -A -A (II)
  • 6 c represents a chromophore or chromophore
  • n an integer of 0 to 20;
  • polyacrylamide polymerization degree 1 to: oligo or polypeptide of LO, oxygen atom or NH;
  • A is a protease derived from Bacillus Licheniformis.
  • a cleavable glutamic acid residue or cysteine residue is a sugar amino acid residue substantially free of a site cleavable by a protease, or
  • glycopeptide residue containing any sugar amino acid without a site cleavable by a protease
  • sugar amino acid residue or the glycopeptide residue is represented by the following formula:
  • R 1 and R 6 are each independently hydrogen, Neu5Ac group or GlcNAc group;
  • R 2 and R 7 are Gal groups
  • R 3 and R 8 are GlcNAc groups
  • R 4 and R 9 are Gal groups
  • R 5 is a GlcNAc group
  • R 10 is a GalNAc group
  • Z 2 and Z 3 are each independently hydrogen or a Fuc group; and n and m are each independently an integer of 0 or more;
  • the bond between R 1 and R 2 is a 2, 3 bond when R 1 is Neu5Ac group, and j81, 3 bond when R 1 is GlcNAc group;
  • R 2 and R 3 are a ⁇ ⁇ , 4 bond
  • R 3 and R 4 are a ⁇ ⁇ , 3 bond
  • R 4 and R 5 are a ⁇ ⁇ , 4 bond
  • R 6 and R 7 are an ⁇ 2,3 bond when R 6 is a Neu5Ac group, and a ⁇ , 3 bond when R 6 is a GlcNA c group;
  • R 7 and R 8 are a ⁇ ⁇ , 4 bond
  • R 8 and R 9 are a ⁇ ⁇ , 3 bond
  • R 9 and R 1G are a ⁇ ⁇ , 3 bond
  • R 5 and R 1G are a ⁇ ⁇ , 6 bond
  • the bond between ⁇ 1 and R 3 is ⁇ 1 , 3 bond;
  • the bond between Z 2 and R 5 is ⁇ 1, 3 bond;
  • the bond between ⁇ 3 and R 8 is ⁇ 1, 3 bond
  • ⁇ is the following formula:
  • the present invention provides the following formula:
  • a N C (— X) — (CH) A -A -A (II)
  • X represents a hydrogen atom, c-c alkyl, c-c aryl or chromophore
  • n an integer of 0 to 20;
  • polyacrylamide polymerization degree 1 to: oligo or polypeptide of LO, oxygen atom or NH;
  • A is a protease derived from Bacillus Licheniformis. A cleavable glutamic acid residue or cysteine residue;
  • A is a sugar amino acid residue substantially free of a site cleavable by a protease, or
  • glycopeptide residue containing any sugar amino acid without a site cleavable by a protease
  • sugar amino acid residue or the glycopeptide residue is represented by the following formula:
  • R 1 is hydrogen, a sialic acid group or a GlcNAc group
  • R 2 is a Gal group
  • R 3 is a GlcNAc group
  • R 4 is a Gal group
  • R 5 is a GlcNAc group
  • R 10 is a GalNAc group
  • R 8 is hydrogen or a sialic acid group
  • R 9 is a Gal group
  • Z 1 and Z 2 are each independently hydrogen or a Fuc group
  • n is an integer greater than or equal to 0;
  • R 1 and R 2 are an a 2, 3 bond when R 1 is a sialic acid group, and a ⁇ ⁇ , 3 bond when R 1 is a GlcNAc group;
  • R 2 and R 3 are a j8 1, 4 bond
  • R 3 and R 4 are 1, 3 bonds
  • R 4 and R 5 are] 3 1, 4 bond;
  • R 8 and R 9 are 2, 3 bonds
  • R 9 and R 1G are a j8 1,3 bond
  • R 5 and R 1G are a j8 1,6 bond
  • the bond between Z 1 and R 3 is an ⁇ ⁇ , 3 bond
  • A is the following formula:
  • the mucin-type glycopeptide having a voralactosamine skeleton provided in the present invention in the formula (I) or ( ⁇ ⁇ ⁇ ), in A, n is an integer of 0 or more, and preferably, n is 0 to Five
  • n is an integer of 4 or less.
  • ⁇ Z 3 may or may not be present, but preferably m is 0 to 5, and when n is 1, Z 1 and Z 2 are sialic acid groups. More preferably, the sugar residue of the glycopeptide is [Chemical 27-2]
  • It can be a sugar residue or a derivative of a sugar residue selected from the group consisting of
  • the present invention provides a composition for a primer for producing a glycoamino acid or glycopeptide comprising the compound described in the above formula (I) or (IV).
  • the glycopeptide containing an amino acid residue that has a ketone residue or an aldehyde residue at the end and can be cleaved by protease is released to the solid phase carrier force, and at the same time, the amino acid side chain protecting group is removed Protect (if the amino acid side chain protecting group is not removed by acid treatment, the protecting group may be separately deprotected by a deprotection reaction);
  • reaction mixture or the mixture obtained by the ether precipitation method is purified by HPLC, and a glycopeptide (sugar chain protector) containing an amino acid residue that has a ketone residue or an aldehyde residue at the end and can be cleaved by a protease is obtained.
  • a glycopeptide sucgar chain protector
  • step 4 is omitted.
  • a polymer carrier is introduced into the reaction solution containing the glycopeptide of 3) and selectively reacted with the glycopeptide;
  • glycopeptide bound to the carrier is purified by gel filtration or dialysis, ultrafiltration, etc .;
  • Synthesis and purification of a polymeric primer from a glycopeptide containing an amino acid residue having a ketone residue or an aldehyde residue at the terminal and thus cleavable by a protease is as follows:
  • the glycopeptide containing an amino acid residue having a ketone residue or an aldehyde residue at the end and cleavable by a protease is released to the solid phase carrier force, and at the same time, the protecting group of the amino acid side chain is removed.
  • Protect if the amino acid side chain protecting group is not removed by acid treatment, the protecting group may be separately deprotected by a deprotection reaction);
  • a polymer carrier is introduced into the reaction solution containing the glycopeptide of 3) to selectively react with the glycopeptide. React;
  • the keto acid or aldehyde acid used in the above step 1) has the following formula:
  • X is a hydrogen atom, c-c alkyl
  • 1 30 c represents 6-c aryl or chromophore
  • n an integer of 0 to 20;
  • (A represents a linker having a length of 1 to 20 methylene chains).
  • the method for producing a glycopeptide of the present invention comprises the following steps:
  • step (B) By reacting the compound obtained in step (A) with a glycosyltransferase in the presence of a sugar nucleotide, the sugar residue is transferred from the sugar nucleotide to the compound, and the sugar chain is elongated.
  • the glycosyltransferase used in the present invention is preferably 13 1, 4-galactose transfer. Enzyme, j8 1,3-Galactosyltransferase, 1,3-Fucose transferase, / 3 1,3-N-Acetyldarcosaminetransferase, 13 1,6-N-Acetyldarcosaminetransferase, ⁇ 2,3-sialyltransferase, ⁇ 2,6-sialyltransferase.
  • the method for producing a glycopeptide of the present invention comprises the following steps:
  • a sugar residue is transferred from the sugar nucleotide to the compound by allowing a glycosyltransferase to act on the compound according to any one of items (4) to (7) in the presence of the sugar nucleotide.
  • (C) a step of allowing a protease to act on a compound in which sugar residues are transferred and sugar chains are elongated. Furthermore, the process of isolating glycopeptide may be included. In this production method, by-products other than the glycopeptide containing the target glycopeptide and the carrier can be easily separated.
  • the method for producing a glycopeptide of the present invention comprises the following steps:
  • a sugar residue is transferred from the sugar nucleotide to the compound by allowing a glycosyltransferase to act on the compound according to any one of items (4) to (7) in the presence of the sugar nucleotide.
  • step (1) Step (ii) Repeating step (1) one or more times to extend the sugar chain;
  • the method for producing a glycopeptide of the present invention comprises the following steps:
  • step (B) The compound obtained in step (A) and an optionally protected aminooxy group, N-alkylaminooxy group, hydrazide group capable of specifically reacting with the ketone residue or aldehyde residue of the compound Reacting with a carrier comprising a functional group selected from the group consisting of: azide group, thiosemicarbazide group, 1,2-dithiol group and cysteine residue;
  • step (C) By allowing a glycosyltransferase to act on the compound obtained in step (B) in the presence of a sugar nucleotide, the sugar residue is transferred from the sugar nucleotide to the compound, and the sugar chain is elongated.
  • (E) a step of allowing a protease to act on a compound in which sugar residues are transferred and sugar chains are elongated.
  • the method for producing a glycopeptide of the present invention comprises the following steps:
  • step (B) The compound obtained in step (A) and an optionally protected aminooxy group, N-alkylaminooxy group, hydrazide group capable of reacting specifically with the ketone residue or aldehyde residue of the compound Reacting with a carrier comprising a functional group selected from the group consisting of: azide group, thiosemicarbazide group, 1,2-dithiol group and cysteine residue;
  • step (C) By allowing a glycosyltransferase to act on the compound obtained in step (B) in the presence of a sugar nucleotide, the sugar residue is transferred from the sugar nucleotide to the compound, and the sugar chain is elongated.
  • Step (D) Step (C) is repeated once or twice or more to extend the sugar chain
  • (F) a step of allowing a protease to act on a compound in which a plurality of sugar residues are transferred and sugar chains are elongated.
  • the method for producing a glycopeptide of the present invention comprises the following steps:
  • step (B) The compound obtained in step (A) and an optionally protected aminooxy group, N-alkylaminooxy group, hydrazide group capable of reacting specifically with the ketone residue or aldehyde residue of the compound , A reaction with a carrier containing a functional group selected from the group consisting of an azide group, a thiosemicarbazide group, a 1,2-dithiol group and a cysteine residue, and simultaneously removing unreacted substances in step (A);
  • the sugar residue is transferred from the sugar nucleotide to the compound by allowing a sugar transferase to act on the compound bound to the carrier obtained in step (B) in the presence of the sugar nucleotide.
  • step (D) A step of allowing protease to act on the compound obtained by extending the sugar chain obtained in step (C) is included.
  • the method for producing a glycopeptide of the present invention comprises the following steps:
  • a peptide solid-phase synthesis is carried out using amino acids, sugar amino acids, and keto acids or aldehyde acids that can be cleaved by a protease, as described in any one of items (1) to (3) Obtaining a compound;
  • step (B) The compound obtained in step (A) and an optionally protected aminooxy group, N-alkylaminooxy group, hydrazide group capable of reacting specifically with the ketone residue or aldehyde residue of the compound , A reaction with a carrier containing a functional group selected from the group consisting of azide group, thiosemicarbazide group, 1,2-dithiol group and cysteine residue, and at the same time, removing unreacted substances in step (A) ;
  • the sugar residue is transferred from the sugar nucleotide to the compound by allowing a sugar transferase to act on the compound bound to the carrier obtained in step (B) in the presence of the sugar nucleotide.
  • Step (D) Step (C) is repeated once or twice or more to extend the sugar chain
  • (F) a step of allowing a protease to act on a compound in which a plurality of sugar residues are transferred and sugar chains are elongated.
  • the method for producing the glycopeptide of the present invention comprises the following steps:
  • a sugar residue is transferred from the sugar nucleotide to the compound by allowing a glycosyltransferase to act on the compound according to any one of items (1) to (3) in the presence of the sugar nucleotide.
  • Step (B) Step (A) is repeated one or more times as necessary to extend the sugar chain as necessary;
  • the method for producing a glycopeptide of the present invention comprises the following steps:
  • a sugar residue is transferred from the sugar nucleotide to the compound by allowing a glycosyltransferase to act on the compound according to any one of items (1) to (3) in the presence of the sugar nucleotide.
  • Step (B) Step (A) is repeated one or more times as necessary to extend the sugar chain as necessary;
  • (C) a compound in which a sugar residue is transferred and a sugar chain is extended, and a protected compound capable of reacting specifically with a ketone residue or an aldehyde residue of the compound, and may be an aminooxy group, N-alkyl Reacting a carrier containing a functional group selected from the group consisting of an aminooxy group, a hydrazide group, an azide group, a thiosemicarbazide group, a 1,2-dithiol group and a cysteine residue;
  • (E) a step of causing a protease to act on a compound in which a sugar residue is transferred and a sugar chain is elongated.
  • the method for producing a glycopeptide of the present invention comprises the following steps:
  • step (B) The compound obtained in step (A) and an optionally protected aminooxy group, N-alkylaminooxy group, hydrazide group capable of reacting specifically with the ketone residue or aldehyde residue of the compound , Azide group, thiosemicarbazide group, 1,2-dithiol group and cysteine Reacting with a soluble carrier containing a functional group selected from the group consisting of residues and removing unreacted substances in step (A) by reprecipitation, gel filtration, or ultrafiltration;
  • step (C) By allowing a glycosyltransferase to act on the compound solublely bound to the carrier obtained in step (B) in the presence of a sugar nucleotide, the sugar residue is transferred from the sugar nucleotide to the compound, A step of obtaining a compound in which a sugar chain is elongated, wherein the sugar nucleotide has a sugar residue selected from the group consisting of a Gal group, a GlcNAc group, a Fuc group and a sialic acid group
  • Step (D) Step (C) is repeated once or twice or more to extend the sugar chain

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Abstract

La présente invention concerne un nouveau composé pouvant servir d'amorce dans la production d'un glycopeptide de type mucine, doté d'un squelette de polylactosamine et utile dans une grande variété d'applications incluant un matériau conçu pour une étude biochimique, un produit pharmaceutique et un aliment, la production dudit composé étant auparavant difficile. L'invention concerne aussi un procédé de production d'un glycopeptide au moyen de l'amorce. L'invention concerne donc : un dérivé de glycopeptide de type mucine dont le nouveau squelette de polylactosamine comporte un groupement aldéhyde ou un groupement cétone au niveau de l'extrémité et également un résidu d'acide aminé clivable à l'aide d'une protéase (c'est-à-dire un composé représenté par la formule (I)) ; un procédé de production simple d'un glycopeptide de type mucine doté d'un squelette de polylactosamine au moyen du dérivé servant d'amorce ; et un procédé de production d'un peptide de type mucine doté d'un squelette de polylactosamine grâce à la synthèse d'un glycopeptide renfermant une séquence peptidique spécifique au moyen d'un acide glycoaminé de type mucine doté d'un squelette polylactosamine qui a été synthétisé préalablement.
PCT/JP2007/057001 2006-03-31 2007-03-29 Glycopeptide de type mucine comportant un squelette de polylactosamine Ceased WO2007114307A1 (fr)

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JP2006100923A JP2009143809A (ja) 2006-03-31 2006-03-31 ポリラクトサミン骨格を有するムチン型糖ペプチド
JP2006-100923 2006-03-31

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2020104486A1 (fr) * 2018-11-19 2020-05-28 Gnubiotics Sciences Sa Compositions de mucines du tractus gastro-intestinal et utilisations associées
US12144842B2 (en) 2018-02-22 2024-11-19 Gnubiotics Sciences Sa Process of preparation of glycan compositions and uses thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005108417A1 (fr) * 2004-05-07 2005-11-17 National Institute Of Advanced Industrial Science And Technology Amorce polymère pour synthèse de peptide de sucre
WO2006030840A1 (fr) * 2004-09-14 2006-03-23 National Institute Of Advanced Industrial Scienceand Technology Procédé pour la synthèse de peptides de type mucine et de glycopeptides apparentés à muc1

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005108417A1 (fr) * 2004-05-07 2005-11-17 National Institute Of Advanced Industrial Science And Technology Amorce polymère pour synthèse de peptide de sucre
WO2006030840A1 (fr) * 2004-09-14 2006-03-23 National Institute Of Advanced Industrial Scienceand Technology Procédé pour la synthèse de peptides de type mucine et de glycopeptides apparentés à muc1

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12144842B2 (en) 2018-02-22 2024-11-19 Gnubiotics Sciences Sa Process of preparation of glycan compositions and uses thereof
WO2020104486A1 (fr) * 2018-11-19 2020-05-28 Gnubiotics Sciences Sa Compositions de mucines du tractus gastro-intestinal et utilisations associées
CN113316586A (zh) * 2018-11-19 2021-08-27 格努生物科学股份公司 来自胃肠道粘蛋白的组合物及其用途

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