WO2023166975A1 - Procédé de production de composé peptidique - Google Patents

Procédé de production de composé peptidique Download PDF

Info

Publication number
WO2023166975A1
WO2023166975A1 PCT/JP2023/005023 JP2023005023W WO2023166975A1 WO 2023166975 A1 WO2023166975 A1 WO 2023166975A1 JP 2023005023 W JP2023005023 W JP 2023005023W WO 2023166975 A1 WO2023166975 A1 WO 2023166975A1
Authority
WO
WIPO (PCT)
Prior art keywords
group
compound
hydrochloride
formula
peptide
Prior art date
Application number
PCT/JP2023/005023
Other languages
English (en)
Japanese (ja)
Inventor
章 大高
Original Assignee
国立大学法人徳島大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 国立大学法人徳島大学 filed Critical 国立大学法人徳島大学
Publication of WO2023166975A1 publication Critical patent/WO2023166975A1/fr

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/10—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length using coupling agents

Definitions

  • the present invention relates to a method for producing a peptide compound.
  • the targets for development of drug candidate compounds are changing day by day, from low-molecular-weight drugs to antibody drugs and then to peptide drugs.
  • the peptide drug can be obtained relatively easily by solid-phase peptide synthesis (SPPS), and is highly promising in that it can target specific cells.
  • SPPS solid-phase peptide synthesis
  • ADCs Antibody Drug Conjugates
  • ADCs Antibody Drug Conjugates
  • the three-dimensional structure of peptides represented by ⁇ -helices, leucine zippers, zinc fingers, coiled coils, etc. is known to be related to the physiological activities of the peptides.
  • an ⁇ -helical structure is formed by interactions between side chains of amino acid residues that constitute a peptide, and stapling that bridges the side chains of multiple amino acids strengthens the ⁇ -helical structure. It has been known.
  • the nucleophilic thiol group present in the side chain of cysteine is involved in alkylation, Michael reaction, disulfide formation, etc., and serves as a bridge for site-specific modification of peptides and proteins. It is known. It is also known that an intramolecular reaction by a nucleophilic thiol group in a peptide is related to cyclization of the peptide, stapling within the peptide, and the like.
  • Non-Patent Document 1 Non-Patent Document 1
  • the linker provided between the antibody and the low-molecular-weight compound poses problems in terms of biocompatibility and metabolism, making it necessary to optimize the linker.
  • the present inventors have found that the side chain of a cysteine residue having a specific protective group and the side chain of a tryptophan residue are in the presence of hydrochloride or metal chloride. It was found to react below to form cross-links. The present inventors also found that such a cross-linking reaction proceeds both intramolecularly within a compound and between molecules of different compounds.
  • the present invention is an invention completed based on these findings, and broadly includes the inventions shown in the following items.
  • R 1 represents a monovalent organic group.
  • R2 represents a divalent organic group.
  • R3 represents a monovalent organic group. Also, R 1 and R 3 may combine with each other to form a ring.
  • R 1 , R 2 and R 3 are the same as above.
  • R 4 represents R 5 —CH 2 — (R 5 represents an alkylcarbonylamino group) or a benzyl group which may have a substituent on the phenyl ring.
  • a production method comprising the step of cross-linking the compound represented by in the presence of hydrochloride or metal chloride.
  • R 1 is a monovalent organic group having a carbonyl group
  • R 2 is a divalent organic group having a carbonyl group and an amino group
  • R 3 is a monovalent organic group having an amino group. 2. The production method according to item 1 above, wherein the organic group is a divalent organic group.
  • Item 3 The production method according to Item 1, wherein each of R 1 , R 2 and R 3 is a peptide residue.
  • Item 4 The production method according to Item 1 above, wherein the substituent on the phenyl ring is an electron-donating group.
  • the electron-donating group is at least one selected from the group consisting of an alkyl group, an alkoxy group, an alkylamino group, an alkylcarbonyl group, an alkylaminocarbonyl group, an alkylcarbonylamino group, a hydroxyl group, an amino group, and a halogen group. 5. The manufacturing method according to item 4 above.
  • the hydrochloride is at least one selected from the group consisting of guanidine hydrochloride, dimethylamine hydrochloride, diisopropylamine hydrochloride, piperidine hydrochloride, tetrabutylammonium chloride, piperazine hydrochloride and morpholine hydrochloride.
  • the manufacturing method according to any one of items 1 to 5 above.
  • Item 7 Any one of items 1 to 5 above, wherein the metal chloride is at least one selected from the group consisting of magnesium chloride, zinc chloride, lithium chloride, iron chloride (III), calcium chloride and nickel chloride. The manufacturing method described in the item.
  • Item 8 The production method according to any one of Items 1 to 7 above, wherein the cross-linking reaction is performed under acidic conditions.
  • R 6 , R 7 , R 8 and R 9 each represent a monovalent organic group.
  • R 8 and R 9 are the same as above.
  • R 10 represents R 5 —CH 2 — (R 5 represents an alkylcarbonylamino group) or a benzyl group which may have a substituent on the phenyl ring.
  • a production method comprising the step of reacting a compound represented by in the presence of a hydrochloride or a metal chloride.
  • Item 10 The production method according to Item 9, wherein R 6 and R 8 are monovalent organic groups having a carbonyl group, and R 7 and R 9 are monovalent organic groups having an amino group.
  • Item 11 The production method according to Item 9, wherein each of R 6 , R 7 , R 8 and R 9 is a peptide residue.
  • Item 12 The production method according to Item 9 above, wherein the substituent on the phenyl ring is an electron-donating group.
  • the electron-donating group is at least one selected from the group consisting of an alkyl group, an alkoxy group, an alkylamino group, an alkylcarbonyl group, an alkylaminocarbonyl group, an alkylcarbonylamino group, a hydroxyl group, an amino group, and a halogen group. 13.
  • the hydrochloride is at least one selected from the group consisting of guanidine hydrochloride, dimethylamine hydrochloride, diisopropylamine hydrochloride, piperidine hydrochloride, tetrabutylammonium chloride, piperazine hydrochloride and morpholine hydrochloride. , the production method according to any one of items 9 to 13 above.
  • Item 15 Any one of items 9 to 13 above, wherein the metal chloride is at least one selected from the group consisting of magnesium chloride, zinc chloride, lithium chloride, iron (III) chloride, calcium chloride and nickel chloride. The manufacturing method described in .
  • Item 16 The production method according to any one of Items 9 to 15 above, wherein the cross-linking reaction is performed under acidic conditions.
  • Item 17 The production method according to any one of Items 9 to 16 above, wherein the cross-linking reaction is further performed in the presence of phenol and/or alkoxybenzene.
  • a novel cross-linking reaction between the side chain of a protected cysteine residue and the side chain of a tryptophan residue can be provided.
  • Such a cross-linking reaction can form a cross-link at a position different from the main chain bond. Therefore, when the cross-linking reaction is carried out within the molecule of the compound, it is possible to form staples between specific amino acid residues, thus making the conformation such as ⁇ -helix more rigid. can be done.
  • the cross-linking reaction is carried out between molecules of different compounds, the backbone of each molecule can be retained, so that while retaining the physiological activity expressed by each compound, direct Each compound can be crosslinked. Therefore, by using the method of the present invention, various peptide drugs can be produced industrially and advantageously.
  • FIG. 1-1 is a diagram showing the results of Example 1.
  • FIG. 1 to 16 are the results of HPLC analysis of the reactants in each sample. * in the figure is a non-peptidic impurity.
  • 1-2 are diagrams showing the results of Example 1.
  • FIG. 1 to 16 are the results of HPLC analysis of the reactants in each sample. * in the figure is a non-peptidic impurity.
  • FIG. 2 is a diagram showing the results of Example 2.
  • FIG. 1 to 5 are the results of HPLC analysis of the reactants in each sample.
  • 3 is a diagram showing the results of Example 3.
  • FIG. 1 to 5 are the results of HPLC analysis of reactants starting from compounds 6 to 10, respectively. 6 is the result of changing the reaction temperature from 20°C to 37°C in the reaction using compound 10 as a raw material.
  • FIG. 1 is the result of HPLC analysis of a reaction product using compound 16 as a raw material.
  • 2 is the result of HPLC analysis of the reaction product using compound 17 as a starting material.
  • 5 is a diagram showing the results of Example 5.
  • FIG. 1 is the result of HPLC analysis of a reaction product using compound 19 as a raw material.
  • 6 is a diagram showing the results of Example 6.
  • FIG. 7 is a diagram showing the results of Example 7.
  • FIG. 1 shows the results of HPLC analysis of compound 24, which is the raw material of compound 25, and 2 shows the results of HPLC analysis of the reaction product of preparing compound 25 using compound 24 as the raw material.
  • 3 is the result of measuring the CD spectra of compound 25 (solid line), compound 26 (dotted line), and control (dashed line).
  • Control is stERAP. * in the figure is a non-peptidic impurity.
  • 8 is a diagram showing the results of Example 8.
  • FIG. 1 is a silver-stained image after the reaction.
  • 2 is a biotin-stained image after the reaction.
  • a in 1 and 2 is a marker and B shows the result of the mixture before the reaction.
  • C shows the results of the reactants after the reaction.
  • FIG. 9 is a diagram showing the results of Example 9.
  • FIG. 1 is the result of HPLC analysis of the reaction product using compound 29 and compound 30 as raw materials.
  • 2 is the result of HPLC analysis of the reaction product using compound 29 and compound 32 as raw materials.
  • 3 is the result of HPLC analysis of the reaction product using compound 35 and compound 30 as raw materials.
  • 4 is the result of HPLC analysis of the reaction product using compound 35 and compound 32 as raw materials.
  • the first manufacturing method of the present invention is the following formula (1)
  • R 1 represents a monovalent organic group.
  • R2 represents a divalent organic group.
  • R3 represents a monovalent organic group. Also, R 1 and R 3 may combine with each other to form a ring.
  • R 1 , R 2 and R 3 are the same as above.
  • R 4 represents R 5 —CH 2 — (R 5 represents an alkylcarbonylamino group) or a benzyl group which may have a substituent on the phenyl ring.
  • R 1 represents a monovalent organic group.
  • a monovalent organic group having a carbonyl group can be mentioned as a preferred embodiment. More specifically, a monovalent organic group in which a carbonyl group is provided so as to form a peptide bond with an amino group adjacent to R 1 is more preferable.
  • Examples of the monovalent organic group having a carbonyl group include a fatty acid group, a monovalent organic group having miniPEG, a peptide residue, and the like.
  • the fatty acid group may be linear or branched, but is preferably linear in view of the ease of production. Although the number of carbon atoms in the above fatty acid is not particularly limited, it can usually be about 12 to 18.
  • MiniPEG is a registered trademark and is a compound represented by 8-amino-3,6-dioxaoctanoic acid.
  • peptide residues are preferred. Peptides constituting such peptide residues may be linear or branched, but are preferably linear in view of ease of production.
  • a peptide residue means an atomic group obtained by removing a hydrogen atom from the N-terminus of a peptide and removing a hydroxyl group from the C-terminus of the peptide.
  • the amino acids constituting the peptide are not limited to the 20 types of narrowly defined amino acids governed by codons, ⁇ -amino acids such as theanine and ornithine, ⁇ -amino acids such as ⁇ -alanine, All structural isomers of ⁇ -amino acids such as ⁇ -aminobutyric acid (GABA) and the like can be employed, and they may be D-amino acids or L-amino acids.
  • GABA ⁇ -aminobutyric acid
  • the number of amino acids contained in the peptide residues exemplified for R 1 is not particularly limited.
  • the number of such amino acids is, for example, usually about 2 to 20, preferably about 2 to 10.
  • the terminal not bonded to the amino group adjacent to R 1 can be protected with an acetyl group, a benzoyl group, a pivaloyl group, or the like.
  • R 2 represents a divalent organic group.
  • Such an organic group is not particularly limited as long as the effects of the present invention are exhibited, and a preferred embodiment is a divalent organic group having a carbonyl group and an amino group. More specifically, a divalent organic group containing a carbonyl group so as to form a peptide bond with the amino group adjacent to R 2 and an amino group so as to form a peptide bond with the carbonyl group adjacent to R 2 It is more preferable to
  • divalent organic group having a carbonyl group and an amino group examples include a fatty acid group having an amino group, a divalent organic group having miniPEG, a peptide residue, and the like.
  • divalent organic groups divalent peptide residues are preferred.
  • a peptide constituting such a peptide residue may be linear or branched, but is preferably linear in view of ease of production.
  • the number of amino acids contained in the peptide residues exemplified for R2 above is not particularly limited as long as the effects of the present invention are not inhibited.
  • the number of such amino acids is, for example, usually about 2 to 20, preferably about 2 to 10, more preferably about 2 to 5.
  • R 3 represents a monovalent organic group.
  • a monovalent organic group is not particularly limited, and a monovalent organic group having an amino group can be mentioned. More specifically, it is more preferable to use a monovalent organic group containing an amino group so as to form a peptide bond with the carbonyl group adjacent to R3 .
  • the monovalent organic group having an amino group examples include a fatty acid group having an amino group, a monovalent amino group having miniPEG, a peptide residue, and the like.
  • peptide residues are preferred.
  • a peptide constituting such a peptide residue may be linear or branched, but preferably linear.
  • the number of amino acids contained in the peptide residues exemplified for R3 is not particularly limited.
  • the number of such amino acids is, for example, usually about 2 to 20, preferably about 2 to 10.
  • the terminal not bonded to the carbonyl group adjacent to R 3 may be protected with a primary amino group, secondary amino group, aromatic amino group, or the like.
  • R 1 and R 3 are bonded to each other to form a ring. That is, the compound represented by Formula (1) can mention the aspect which forms a cyclic peptide.
  • Other embodiments of the compound represented by formula (1) include, for example, an embodiment containing a disulfide bond via a cysteine residue contained in formula (1), an oxoacid and hydroxyl Examples include an aspect containing an ester bond formed with a group.
  • R 4 in the above formula (2) represents R 5 —CH 2 — or a benzyl group which may have a substituent on the phenyl ring.
  • R 5 represents an alkylcarbonylamino group.
  • the alkylcarbonylamino group is a group represented by the formula below (R 11 represents an alkyl group) and is also called an alkanoylamino group.
  • the substituent on the phenyl ring is not particularly limited. Examples of such substituents include electron-donating groups.
  • the electron-donating group is not particularly limited as long as it does not inhibit the effects of the present invention. , hydroxyl group, amino group and halogen group.
  • alkyl group examples include methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, s-butyl group, t-butyl group, 1-ethylpropyl group and n-pentyl group. , a neopentyl group, an n-hexyl group, an isohexyl group, a 3-methylpentyl group, and other linear or branched alkyl groups having 1 to 6 carbon atoms (especially 1 to 4 carbon atoms). Among these alkyl groups, a methyl group is preferred.
  • alkoxy group examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy, n-pentyloxy, neopentyloxy, and n-hexyloxy groups.
  • Linear or branched alkoxy groups having 1 to 6 carbon atoms (especially 1 to 4 carbon atoms) can be mentioned.
  • a methoxy group and the like are preferable.
  • alkylamino group examples include methylamino group, ethylamino group, n-propylamino group, isopropylamino group, n-butylamino group, isobutylamino group, s-butylamino group, t-butylamino group, 1 -ethylpropylamino group, n-pentylamino group, neopentylamino group, n-hexylamino group, isohexylamino group, monoalkylamino groups such as 3-methylpentylamino group; dimethylamino group, diethylamino group, di- Linear or branched alkylamino groups having 1 to 6 carbon atoms (especially 1 to 4 carbon atoms) such as dialkylamino groups such as n-propylamino group can be mentioned.
  • alkylcarbonyl group examples include methylcarbonyl group, ethylcarbonyl group, n-propylcarbonyl group, isopropylcarbonyl group, n-butylcarbonyl group, isobutylcarbonyl group, s-butylcarbonyl group, t-butylcarbonyl group, 1 -An alkyl moiety such as an ethylpropylcarbonyl group, n-pentylcarbonyl group, neopentylcarbonyl group, n-hexylcarbonyl group, isohexylcarbonyl group, 3-methylpentylcarbonyl group has 1 to 6 carbon atoms (especially 4), for example, a linear or branched alkylcarbonyl group.
  • alkylaminocarbonyl groups include methylaminocarbonyl, ethylaminocarbonyl, n-propylaminocarbonyl, isopropylaminocarbonyl, n-butylaminocarbonyl, isobutylaminocarbonyl and s-butylaminocarbonyl groups.
  • alkyl moiety such as dialkylaminocarbonyl group such as dimethylaminocarbonyl group, diethylaminocarbonyl group and di-n-propylaminocarbonyl group has 1 to 6 carbon atoms (especially 1 to 4 carbon atoms) Linear or branched alkylaminocarbonyl groups and the like can be mentioned.
  • alkylcarbonylamino group examples include methylcarbonylamino group, ethylcarbonylamino group, n-propylcarbonylamino group, isopropylcarbonylamino group, n-butylcarbonylamino group, isobutylcarbonylamino group and s-butylcarbonylamino group.
  • t-butylcarbonylamino group 1-ethylpropylcarbonylamino group, n-pentylcarbonylamino group, neopentylcarbonylamino group, n-hexylcarbonylamino group, isohexylcarbonylamino group, 3-methylpentylcarbonylamino group, etc.
  • a linear or branched alkylcarbonylamino group having 1 to 6 carbon atoms (especially 1 to 4 carbon atoms) in the alkyl moiety of is mentioned.
  • halogen group examples include fluorine, chlorine, bromine, and iodine.
  • the hydrochloride used in the first production method of the present invention is not particularly limited as long as it can exhibit the effects of the present invention. Specifically, guanidine hydrochloride, dimethylamine hydrochloride, diisopropylamine hydrochloride, piperidine Hydrochloride, tetrabutylammonium chloride (nBu 4 NCl), piperazine hydrochloride, morpholine hydrochloride and the like can be mentioned.
  • guanidine hydrochloride dimethylamine hydrochloride, diisopropylamine hydrochloride, piperidine hydrochloride, piperazine hydrochloride, or morpholine hydrochloride is preferred, and guanidine hydrochloride, diisopropylamine hydrochloride, or piperidine hydrochloride is preferred. Most preferred.
  • the amount of hydrochloride used above can be appropriately set within a wide range.
  • the hydrochloride can be used in an amount of about 1 to 4 mol, preferably about 2 to 4 mol, relative to the molar amount of the compound represented by formula (2).
  • the metal chloride used in the first production method of the present invention is not particularly limited as long as it can exhibit the effects of the present invention, and specifically, magnesium chloride, zinc chloride, lithium chloride, iron (III) chloride. , calcium chloride, nickel chloride and the like. Among these metal chlorides, magnesium chloride is preferred.
  • the amount of metal chloride used can be set appropriately from a wide range.
  • the metal chloride can be used in an amount of about 25 to 40 molar amounts. A molar amount is preferred.
  • R 4 in formula (2) represents R 5 —CH 2 — (R 5 is an alkylcarbonylamino group), it is preferred to use a metal chloride.
  • the reaction conditions of the first production method of the present invention are not particularly limited as long as the effects of the present invention are exhibited, but for example, it is preferably carried out under acidic conditions.
  • the reaction system contains acids such as trifluoromethanesulfonic acid, methanesulfonic acid, trifluoroacetic acid, trimethylsilyl trifluoromethanesulfonate, 1-butyl-1-methylpyrrolidinium trifluoromethanesulfonic acid, and acetic acid. It is possible to mention that
  • the reaction solvent in the first production method of the present invention is not particularly limited as long as it does not inhibit the reaction.
  • trifluoroacetic acid and the like can be mentioned.
  • trifluoroacetic acid can also be used as an acid.
  • the reaction temperature in the first production method of the present invention is not particularly limited.
  • the reaction temperature can be about 20 to 70°C, preferably about 30 to 50°C.
  • the reaction time of the first production method of the present invention varies depending on the reaction temperature, and although it cannot be generalized, it can be, for example, about 1 to 5 hours.
  • the cross-linking reaction product (compound of formula (1)) obtained by the first production method of the present invention can be subjected to isolation and purification means to obtain a highly pure target product.
  • Specific isolation and purification means can be appropriately combined with known means such as column chromatography such as HPLC, thin layer chromatography, recrystallization, reprecipitation, distillation, and solvent extraction.
  • the second manufacturing method of the present invention is the following formula (3)
  • R 6 , R 7 , R 8 and R 9 each represent a monovalent organic group.
  • R 8 and R 9 are the same as above.
  • R 10 represents R 5 —CH 2 — (R 5 represents an alkylcarbonylamino group) or a benzyl group which may have a substituent on the phenyl ring. ] and a compound represented by in the presence of hydrochloride or metal chloride.
  • R 6 , R 7 , R 8 and R 9 in formulas (3), (4) and (5) above are each monovalent organic groups. These monovalent organic groups can be the same as the monovalent organic groups specifically described in the first production method of the present invention.
  • R 10 in the above formula (5) may have a substituent on R 5 —CH 2 — or the phenyl ring, like R 4 specifically explained in the first production method of the present invention. It shows a good benzyl group.
  • the usage ratio of the compound represented by formula (4) and the compound represented by formula (5) in the second production method of the present invention is not particularly limited.
  • the molar ratio of the former to the latter can be about 1:0.5-2, preferably about 1:0.8-1.5, more preferably 1:0.9-1.
  • a ratio of about 1 can be used.
  • the hydrochloride in the second production method of the present invention can be the same as the hydrochloride specifically explained in the first production method of the present invention.
  • the metal chloride in the second production method of the present invention can be the same as the metal chloride specifically explained in the first production method of the present invention.
  • R 10 in formula (5) represents R 5 —CH 2 — (R 5 is an alkylcarbonylamino group)
  • R 5 is an alkylcarbonylamino group
  • reaction conditions in the cross-linking reaction of the second production method of the present invention are also preferably carried out under acidic conditions, similar to the reaction conditions of the first production method of the present invention.
  • the means for providing acidic conditions can be as described in detail in the first production method of the present invention.
  • the reaction solvent in the second production method of the present invention can be the same as the solvent in the cross-linking reaction in the first production method of the present invention. More specifically, when R 10 is a benzyl group which may have a substituent on the phenyl ring, it is preferred to use methanesulfonic acid in addition to trifluoroacetic acid. When R 10 is R 5 —CH 2 — (R 5 is an alkylcarbonylamino group), the ionic liquid 1-butyl-1-methylpyrrolidinium trifluoromethanesulfonic acid (BMPy ⁇ OTf) can also be used as a solvent.
  • BMPy ⁇ OTf 1-butyl-1-methylpyrrolidinium trifluoromethanesulfonic acid
  • the cross-linking reaction can also be carried out in the presence of phenol and/or alkoxybenzene.
  • the reaction can proceed more efficiently.
  • the alkoxybenzene is not particularly limited as long as the effects of the present invention are exhibited, and examples include anisole (methoxybenzene), ethoxybenzene, propoxybenzene, and butoxybenzene. Among these alkoxybenzenes, anisole and ethoxybenzene are preferred, and anisole is particularly preferred. In addition, the said alkoxybenzene can be used individually or in combination of 2 or more types.
  • the amount of the phenol and/or alkoxybenzene used can be used in a wide range as long as it does not inhibit the effects of the present invention. can be done.
  • the reaction temperature in the cross-linking reaction of the second production method of the present invention can be the same as the reaction temperature of the first production method of the present invention.
  • the reaction time in the first production method of the present invention can be the same as the reaction time in the first production method of the present invention.
  • R 10 is a benzyl group which may have a substituent on the phenyl ring
  • the reaction temperature may be in the range of 0 to 70°C, which is lower than the reaction temperature of the first production method. can.
  • the compound obtained by the second production method of the present invention (compound of formula (3)) is subjected to isolation and purification means to obtain a highly pure target product.
  • isolation and purification means can be appropriately combined with known means such as column chromatography such as HPLC, thin layer chromatography, recrystallization, reprecipitation, distillation, and solvent extraction.
  • MS analysis in the examples described later was performed by Waters MICROMASS (registered trademark) LCT PREMIERTM (ESI-TOF) or LC-MS (Shimadzu, Japan, Prominence-I LC-2030, LCMS-2020). did.
  • HPLC analyzes in the examples below were performed using a HITACHI L-7150 L-2400 detector or a Waters Alliance 2695 Separations Module with ELS 2420 System using a Cosmosil 5C18-AR-II analytical column (Nacalai Tesque, 4 .6 ⁇ 250 mm, flow rate 1.0 mL ⁇ min ⁇ 1 ) and Cosmosil 5C18-AR-II semi-preparative columns (Nacalai Tesque, 10 ⁇ 250 mm, flow rate 3.0 mL ⁇ min ⁇ 1 ).
  • NMR measurement in the examples described later uses Bruker AV400N (400 MHz) or Bruker AV500 (500 MHz) for 1 H, and Bruker AV400N (100 MHz) or Bruker AV500 (125 MHz) for 13 C. did.
  • CD spectra in the examples described later were measured by a conventional method using a JASCO J-1500 CD spectrometer unless otherwise specified.
  • Fmoc-SPPS Fmoc SPPS was performed according to the following protocol.
  • Cys(Acm)(O)-containing peptide was obtained by subjecting the Cys(Acm)-containing peptide to an oxidation reaction after obtaining the Cys(Acm)-containing peptide by a conventional Fmoc solid-phase synthesis method.
  • Example 1 In the following reaction scheme, the following compound 2 was obtained by reacting the following compound 1 as a raw material under various conditions. Table 1 shows specific conditions. The reaction temperature in this reaction was 4° C. and the reaction time was 3 hours. In this reaction, the solvent used was TFA, which acted as an acid. Compound 1 was produced by the peptide synthesis method described above.
  • GAL indicates residues of a peptide consisting of glycine-alanine-leucine in order from the N-terminal side.
  • the carbonyl group adjacent to G (glycine) and the amino group of the glycine are peptide-bonded
  • the amino group adjacent to L (leucine) and the carbonyl group of leucine are peptide-bonded.
  • R indicates an arginine residue.
  • TFMSA trifluoromethanesulfonic acid
  • MSA methanesulfonic acid
  • nBu 4 NCl tetrabutylammonium chloride
  • FIG. 1 shows the HPLC chart of the reaction product obtained in this reaction.
  • MS data and NMR data of the compound 2 are as follows.
  • Example 2 In the reaction scheme below, compound 3 and compound 4 were reacted with compound 5 under various conditions shown in Table 2 below. The reaction temperature in this reaction was 4° C. and the reaction time was 3 hours. TFA was used as solvent in this reaction. For compound 3, the carbonyl group of L-tryptophan was converted to a methoxy group by a conventional method in the presence of thionyl chloride. Compound 4 was produced by the peptide synthesis method described above.
  • Ac- represents an acetyl group.
  • FIG. 2 shows the HPLC chart of the reaction product obtained in this reaction. Moreover, the MS data of the obtained compound 5 are as follows. ⁇ Compound 5> LRMS ( ESI -TOF) m/z: [M+H] + calcd for C25H37N5O6S 534.2 , found 533.9.
  • Example 3 In the reaction scheme below, the following compounds 6 to 10 were reacted under the conditions of 1M MSA and 4M guanidine hydrochloride as starting materials. The reaction temperature in this reaction was 20° C., and the reaction time was 3 hours. TFA was used as solvent in this reaction. In addition, the reaction was similarly performed at 37° C. instead of 20° C. using compound 10 as a starting material. Compounds 6-10 were made by the peptide synthesis method described above.
  • the MS data of the prepared compounds 6 to 10 are as follows. ⁇ Compound 6> LRMS (ESI-TOF ) m/z: [M+2H] + calcd for C66H102N22O15S 737.4 , found 737.1. ⁇ Compound 7> LRMS (ESI-TOF) m/z: [ M+2H] + calcd for C66H107N21O15S 732.9 , found 732.7. ⁇ Compound 8> LRMS (ESI - TOF) m/z: [M+2H] + calcd for C69H104N20O15S 742.4 , found 742.2.
  • Ac- represents an acetyl group
  • GAL represents a peptide residue consisting of glycine-alanine-leucine in order from the N-terminal side
  • GHRAL represents glycine-histidine-arginine-alanine- in order from the N-terminal side.
  • GKRAL indicates a peptide residue consisting of glycine-lysine-arginine-alanine-leucine in order from the N-terminal side
  • GFRAL indicates glycine-phenylalanine-arginine-alanine- in order from the N-terminal side
  • GMRAL indicates glycine-methionine-arginine-alanine- in order from the N-terminal side
  • Peptide residues consisting of leucine are indicated.
  • the carbonyl group adjacent to the N-terminal G (glycine) residue and the primary amino group of the glycine residue are peptide-bonded, and the amino acid adjacent to the C-terminal L (leucine) residue A peptide bond is formed between the group and the carbonyl group of the leucine residue.
  • RG indicates residues of a peptide consisting of arginine-glycine in order from the N-terminus.
  • the carbonyl group adjacent to the R (arginine) residue and the amino group of arginine are peptide-bonded, and the G (glycine) residue has a C-terminal amide structure.
  • FIG. 3 shows the HPLC chart of the reaction product obtained in the above reaction.
  • the MS data of Compounds 11 to 15 obtained are as follows. ⁇ Compound 11> LRMS ( ESI-TOF) m/z: [M+2H] + calcd for C58H92N22O13S 668.4 , found 668.2 . ⁇ Compound 12> LRMS (ESI-TOF ) m/z: [M+2H] + calcd for C58H97N21O13S 663.9 , found 663.7 .
  • cross-linking reaction temperature was 37°C more efficiently than 20°C.
  • Example 4 In the reaction scheme below, compounds 16 and 17 below were reacted under the conditions of 1 M MSA and 4 M guanidine hydrochloride. The reaction temperature in this reaction was 20° C., and the reaction time was 3 hours. TFA was used as solvent in this reaction. These compounds 16 and 17 were prepared by the peptide synthesis method described above.
  • Ac- represents an acetyl group
  • GA represents a peptide residue consisting of glycine-alanine in order from the N-terminus
  • GALRA is a peptide consisting of glycine-alanine-leucine-arginine-alanine in order from the N-terminus. Residues are indicated.
  • the carbonyl group adjacent to the G (glycine) residue on the N-terminal side and the amino group of the glycine residue are peptide-bonded, and the amino group adjacent to the A (alanine) residue on the C-terminal side A peptide bond is formed between the group and the carbonyl group of the alanine residue.
  • R represents an arginine residue.
  • FIG. 4 shows the HPLC chart of the reaction product obtained in the above reaction.
  • MS data of the obtained compounds 18 and 19 are as follows. ⁇ Compound 18> LRMS ( ESI -TOF) m/z: [ M +H] + calcd for C27H39N10O6S 631.3, found 631.3 . ⁇ Compound 19> LRMS (ESI-TOF ) m /z: [ M+2H] + calcd for C42H68N16O9S 486.3, found 486.3.
  • Example 5 In the reaction scheme below, the following compound 20 was used as a starting material and reacted under the conditions of 1M MSA and 4M guanidine hydrochloride. The reaction temperature in this reaction was 25° C. and the reaction time was 30 minutes. TFA was used as solvent in this reaction. This compound 19 was prepared by the peptide synthesis method described above.
  • Ac- represents an acetyl group
  • GYRAL represents a peptide residue consisting of glycine-tyrosine-arginine-alanine-leucine in order from the N-terminus
  • GAL consists of glycine-alanine-leucine in order from the N-terminus.
  • Peptide residues are indicated.
  • the carbonyl group adjacent to the G (glycine) residue and the amino group of the glycine residue are peptide-bonded, and the primary amino group adjacent to the L (leucine) residue and the leucine residue A peptide bond is formed with the carboxynyl group.
  • RG represents a peptide residue consisting of glycine-arginine in order from the N-terminus.
  • the carbonyl group adjacent to the R (arginine) residue and the amino group of the arginine residue are peptide-bonded, and the G (glycine) residue has a C-terminal amide structure.
  • compound 20 which has a cysteine residue and a tryptophan residue protected by a methylcarbonylamino group contained in the peptide, under acidic conditions in the presence of hydrochloride (guanidine hydrochloride), the cysteine residue and tryptophan residues.
  • hydrochloride guanidine hydrochloride
  • Example 6 As shown in the scheme below, compound 21 prepared by the peptide synthesis method described above was converted to N-methylpyrrolidone (NMP) containing tripyrrolidinophosphonium hexafluorophosphate (e.g., PyBOP) and N,N-diisopropylethylamine (DIEA). ) at room temperature for 3 hours to obtain a cyclic peptide of compound 22.
  • NMP N-methylpyrrolidone
  • DIEA N,N-diisopropylethylamine
  • this compound 22 was reacted under the conditions of 1M MSA and 4M guanidine hydrochloride.
  • the reaction temperature in this reaction was 4° C. and the reaction time was 3 hours.
  • TFA was used as solvent in this reaction.
  • NPI indicates a peptide residue consisting of asparagine-proline-isoleucine in order from the N-terminus.
  • the carbonyl group adjacent to the N (asparagine) residue and the amino group of the asparagine residue are peptide-bonded, and the primary amino group adjacent to the I (isoleucine) residue and the isoleucine residue A peptide bond is formed with the carbonyl group.
  • GI indicates a peptide residue consisting of glycine-isoleucine in order from the N-terminus.
  • the carbonyl group adjacent to the G (glycine) residue and the primary amino group of the glycine residue are peptide-bonded
  • the primary amino group adjacent to the I (isoleucine) residue and the carbonyl of the isoleucine residue are group is peptide-bonded.
  • Example 7 In the reaction scheme below, compound 24 was used as a starting material and reacted under the conditions of 1M MSA and 4M guanidine hydrochloride. The reaction temperature in this reaction was 4° C. and the reaction time was 3 hours. TFA was used as solvent in this reaction. Compound 24 was made by the peptide synthesis method described above.
  • Ac- represents an acetyl group
  • SDL represents a peptide residue consisting of serine-aspartic acid-leucine in order from the N-terminus.
  • the carbonyl group adjacent to the S (serine) residue and the amino group of the serine residue are peptide-bonded
  • the amino group adjacent to the L (leucine) residue and the carbonyl group of the leucine residue is a peptide bond.
  • LQLRQR indicates a peptide residue composed of leucine-glutamine-leucine-arginine-glutamine-arginine in order from the N-terminus.
  • the carbonyl group adjacent to the L (leucine) residue on the N-terminal side and the amino group of the leucine residue are peptide-bonded, and the R (arginine) on the C-terminal side has a C-terminal amide structure.
  • Compound 24 is an analog of the amino acid sequence of stERAP, a breast cancer inhibitory peptide, and the norleucine residue in compound 24 corresponds to the methionine residue of stERAP.
  • stERAP a bisamide-mediated cross-linking is performed between a glutamic acid residue provided between a methionine residue and a serine residue and a glutamine residue provided between leucine residues. It is Here, it was assumed that formation of an intramolecular cross-link between the cysteine residue and the tryptophan residue in compound 24 would give a compound having the same three-dimensional structure as stERAP, like compound 25.
  • FIG. 7 shows the HPLC chart of the reaction product obtained in the above reaction and the results of the CD spectrum after the reaction.
  • Example 8 0.01 mM tratusuzumab (Chugai Pharmaceutical Co., Ltd.) and compound 27 represented by the following formula were mixed and reacted in 0.1% TFA containing 30 mM magnesium chloride and 5% water at 37° C. for 24 hours.
  • the reaction system also contained 1-butyl-1-methylpyrrolidinium trifluoromethanesulfonic acid.
  • Compound 27 was made by the peptide synthesis method described above.
  • a in the above compound 27 is a group shown below, in compound 27 the carbonyl group adjacent to A and the amine group of A are bonded to the peptide, and the amine group adjacent to A and the carbonyl group of A bound to peptides.
  • Example 9 In the reaction scheme below, compounds 29, 30, 32, and 35 were reacted under conditions of 1M MSA and 4M diisopropylamine hydrochloride as starting materials. The reaction temperature in this reaction was 4° C. and the reaction time was 3 hours. TFA was used as solvent in this reaction.
  • H at the N-terminus represents a histidine residue
  • EGTFFTSDVSSYLEGQAAKEFIA is glutamic acid-glycine-threonine-phenylalanine-threonine-serine-aspartic acid-valine-serine-serine-tyrosine-leucine in order from the N-terminal side.
  • LVRGRG represents a peptide residue consisting of leucine-valine-arginine-glycine-arginine-glycine in order from the N-terminus.
  • the carbonyl group adjacent to the N-terminal L (leucine) residue and the amino group of the leucine are peptide-bonded.
  • A is the same as A above, and in compound 30, the carbonyl group adjacent to A) and the amine group of A are bonded to the peptide, and the amine group adjacent to A and the carbonyl of A group is attached to the peptide.
  • A3 is a group shown below. In compound 32, the carbonyl group adjacent to A3 and the amine group of A3 are bonded to the peptide, and the amine group adjacent to A3 and the amine group of A3 are bonded to the peptide . A carbonyl group is attached to the peptide.
  • FIG. 9 shows the HPLC chart of the reaction product obtained in the above reaction.
  • MS data of the obtained compounds 31, 33, 36 and 37 are as follows.
  • compounds 29 and 35 are bioactive peptides, and lipid structures are being actively bound to them.
  • Compounds 30 and 32 are expected to increase the in vivo stability of the peptide and suppress renal excretion by introducing a lipid structure.
  • Compounds 31, 33, 36 and 37 in which a cysteine sulfoxide protected by a methoxybenzyl group is used to introduce a lipid structure onto tryptophan via intermolecular cross-linking, exhibit more effective physiological activity in vivo. be able to.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)

Abstract

La présente invention permet de renforcer une structure d'hélice α impliquée dans la bioactivité d'un peptide. La présente invention permet de conduire à une réaction entre une chaîne latérale d'un résidu tryptophane et une chaîne latérale d'un résidu cystéine pourvu d'une protection spécifique, en présence d'un chlorhydrate ou d'un chlorure métallique.
PCT/JP2023/005023 2022-03-01 2023-02-14 Procédé de production de composé peptidique WO2023166975A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2022031052 2022-03-01
JP2022-031052 2022-03-01

Publications (1)

Publication Number Publication Date
WO2023166975A1 true WO2023166975A1 (fr) 2023-09-07

Family

ID=87883403

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2023/005023 WO2023166975A1 (fr) 2022-03-01 2023-02-14 Procédé de production de composé peptidique

Country Status (1)

Country Link
WO (1) WO2023166975A1 (fr)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20120214968A1 (en) * 2011-02-17 2012-08-23 Baosheng Liu Preparation of phalloidin and its derivatives

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20120214968A1 (en) * 2011-02-17 2012-08-23 Baosheng Liu Preparation of phalloidin and its derivatives

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
KOBAYASHI DAISHIRO, KOHMURA YUTAKA, HAYASHI JUNYA, DENDA MASAYA, TSUCHIYA KOICHIRO, OTAKA AKIRA: "Copper(II)-mediated C-H sulphenylation or selenylation of tryptophan enabling macrocyclization of peptides", CHEMICAL COMMUNICATIONS, ROYAL SOCIETY OF CHEMISTRY, UK, vol. 57, no. 82, 14 October 2021 (2021-10-14), UK , pages 10763 - 10766, XP093087595, ISSN: 1359-7345, DOI: 10.1039/D1CC04856B *
KOBAYASHI DAISHIRO, KOHMURA YUTAKA, SUGIKI TOSHIHIKO, KURAOKA EISUKE, DENDA MASAYA, FUJIWARA TOSHIMICHI, OTAKA AKIRA: "Peptide Cyclization Mediated by Metal‐Free S‐Arylation: S‐Protected Cysteine Sulfoxide as an Umpolung of the Cysteine Nucleophile", CHEMISTRY - A EUROPEAN JOURNAL, JOHN WILEY & SONS, INC, DE, vol. 27, no. 56, 7 October 2021 (2021-10-07), DE, pages 14092 - 14099, XP093087593, ISSN: 0947-6539, DOI: 10.1002/chem.202102420 *
KOBAYASHI DAISHIRO, KURAOKA EISUKE, HAYASHI JUNYA, YASUDA TAKUMA, KOHMURA YUTAKA, DENDA MASAYA, HARADA NORIO, INAGAKI NOBUYA, OTAK: "S-Protected Cysteine Sulfoxide-Enabled Tryptophan-Selective Modification with Application to Peptide Lipidation", ACS MEDICINAL CHEMISTRY LETTERS, AMERICAN CHEMICAL SOCIETY, US, vol. 13, no. 7, 14 July 2022 (2022-07-14), US , pages 1125 - 1130, XP093087597, ISSN: 1948-5875, DOI: 10.1021/acsmedchemlett.2c00161 *
TAISHIRO KOBAYASHI, YUTAKA MITSUMURA, EISUKE KURAOKA, MASAYA DENDA, AKIRA OTAKA: "27V03-pm03S Development of the thioether-forming reaction between Cys and Trp using S-protected cysteine sulfoxide", ABSTRACTS OF ANNUAL MEETING OF PHARMACEUTICAL SOCIETY OF JAPAN (CD-ROM), PHARMACEUTICAL SOCIETY OF JAPAN, JP, vol. 141, 3 May 2021 (2021-05-03) - 29 March 2021 (2021-03-29), JP , pages 27V03 - pm03S, XP009549254, ISSN: 0918-9823 *

Similar Documents

Publication Publication Date Title
US20070207947A1 (en) Stabilized Peptides
CN115925790A (zh) 用于合成α4β7肽拮抗剂的方法
JP6500105B2 (ja) ペプチドが結合された安定したアスコルビン酸誘導体、その製造方法、及びそれを含む化粧料組成物
WO2019069978A1 (fr) Procédé de production de composé peptidique
WO2011098999A2 (fr) Préparation chimique de thioesters d'ubiquitine et modifications de ces derniers
CN111040020B (zh) 一种烯烃硫醚类订书肽及其制备方法与应用
EP3932899A1 (fr) Procédé de production d'un composé peptidique, réactif formant un groupe protecteur et composé hydrocarboné aromatique polycyclique fusionné
JP7372320B2 (ja) 化合物又はその塩、その調製方法および応用
WO2023166975A1 (fr) Procédé de production de composé peptidique
WO2020201816A1 (fr) Composés cycliques et leurs procédés de fabrication et d'utilisation
Zhang et al. Influence of α-methylation in constructing stapled peptides with olefin metathesis
US6277958B1 (en) Method for preparing peptide thiol ester
CA2534224C (fr) Preparation de peptides somatostatine
CN105622424B (zh) 化合物及其制备方法和应用
EP2226314A1 (fr) Agents de reticulation
EP4083054A1 (fr) Procédé de production d'oligomère d'anp dans un procédé en solution
WO2006105199A2 (fr) Compositions et methodes de synthese d'un peptide et d'un conjugue apparente
WO2022090448A1 (fr) Nouveaux réactifs d'acylation
Jiang et al. Synthesis of Thiol‐Containing Oligopeptides via Tandem Activation of γ‐Thiolactones by Silver‐DABCO Pair
Matera-Witkiewicz et al. Short-chain oligopeptides with copper (II) binding properties: The impact of specific structural modifications on the copper (II) coordination abilities
WO2020210535A9 (fr) Macrocyclisation et multimérisation modulables de peptides et de protéines par cycloadditions diels-alder
JP5947821B2 (ja) セレウリドおよびその誘導体の製造方法、セレウリド製造の為の中間体ならびにセレウリド誘導体
AU2014234400A1 (en) Synthesis of hydantoin containing peptide products
CA2907521C (fr) Synthese de produits peptidiques contenant un imide cyclique
Zavitsanos et al. Interaction of Cu (ii) and Ni (ii) with the 63–93 fragment of histone H2B

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23763236

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2024504597

Country of ref document: JP

Kind code of ref document: A