Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07F—ACYCLIC, CARBOCYCLIC OR HETEROCYCLIC COMPOUNDS CONTAINING ELEMENTS OTHER THAN CARBON, HYDROGEN, HALOGEN, OXYGEN, NITROGEN, SULFUR, SELENIUM OR TELLURIUM
  • C07F7/00—Compounds containing elements of Groups 4 or 14 of the Periodic Table
  • C07F7/02—Silicon compounds
  • C07F7/08—Compounds having one or more C—Si linkages
  • C07F7/18—Compounds having one or more C—Si linkages as well as one or more C—O—Si linkages
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
  • C07K1/02—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length in solution
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07K—PEPTIDES
  • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
  • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
  • C07K5/06—Dipeptides

Definitions

• the present invention relates to a method for producing a novel silane-containing fused ring dipeptide compound.
• Non-patent Documents 1 to 3 Non-patent Documents 1 to 3
• amidation which is the most important step in peptide synthesis
• the cost of disposal and purification of by-products accounts for most of the costs of peptide synthesis and is one of the biggest barriers to the development of this field.
• Patent Document 4 technology to synthesize peptides consisting of various amino acid residues with high efficiency and high selectivity by deprotection
• Patent Document 7 technology to synthesize peptides consisting of various amino acid residues with high efficiency and high selectivity by deprotection
• Patent Document 7 technology to synthesize peptides consisting of various amino acid residues with high efficiency and high selectivity by deprotection
• Patent Document 7 technology to synthesize peptides consisting of various amino acid residues with high efficiency and high selectivity by deprotection.
• the method for producing the silane-containing fused ring dipeptide compound disclosed by the present inventors in Patent Document 8 includes a condensation reaction between an unprotected amino acid as an electrophilic species and an amino acid ester as a nucleophilic species, ester deprotection, and This method consists of a cyclization reaction.
• This method does not require protection of the N-terminus of the electrophilic amino acid, and also allows automatic deprotection of the nucleophilic amino acid ester in the system, allowing additional peptides to be added to both ends of the generated peptide. It has the advantage of being able to carry out an elongation reaction.
• an amino acid ester C-terminally protected amino acid
• R 11 , R 12 , R 13 , R 21 , and R 22 are each independently a hydrogen atom, a halogen atom, a hydroxyl group, a carboxyl group, a nitro group, a cyano group, or a thiol group, or one or more substituents
• R a1 and R a2 each independently represent a monovalent aliphatic hydrocarbon group or an aromatic hydrocarbon group which may have one or more substituents.
• R b1 , R b2 , and R b3 are each independently a hydrogen atom, a halogen atom, or a monovalent aliphatic hydrocarbon group or an aromatic carbonization group which may have one or more substituents.
• n b represents an integer of 1 or 2
• Z b is an amino group, carbonylamino group, acetamido group, which may have one or more substituents, or 5 containing one or more nitrogen atoms as ring constituent atoms.
• ⁇ represents a 10-membered monovalent heterocyclic group
• Z b represents a nitrogen-containing divalent linking group.
• R a3 represents a hydrogen atom, a carboxyl group, a hydroxyl group, a monovalent aliphatic hydrocarbon group that may have one or more substituents, an aromatic hydrocarbon group, or a heterocyclic group; , where, in the case of a monovalent aliphatic hydrocarbon group, aromatic hydrocarbon group, or heterocyclic group, it may be bonded to the nitrogen atom via a linking group, Alternatively, R a1 and R a3 are bonded to each other to form a heterocycle which may have one or more substituents together with the carbon atom to which R a1 is bonded and the nitrogen atom to which R a3 is bonded.
• a a1 and A a2 each independently represent a divalent aliphatic hydrocarbon group having 1 to 3 carbon atoms which may have one or more substituents, p a1 and p a2 each independently represent 0 or 1, m a is an integer of 1 or more and represents the number of structural units represented by the structure in [ ]. However, when m is 2 or more, the plurality of structural units represented by the structures in [ ] may be the same or different.
• PG a , R a1 , R a2 , R a3 , A a1 , A a2 , p a1 , p a2 , and m a represent groups having the same definition as the groups with the same symbols in the formula (Ra), R 111 , R 112 , R 113 , R 211 , R 212 , R x11 , R x12 , PG x1 , R 121 , R 122 , R 123 , R 221 , R 222 , R x21 , and R x22 are represented by the formula ( Represents a group with the same definition as the group with the same symbol in P4).
• Non-Patent Document 9 Non-Patent Document 9
• Patent Document 8 This condensed ring dipeptide compound has both the N-terminus and C-terminus of the dipeptide protected by silicon, which is easily deprotected, so it can be easily deprotected and can be used as both a nucleophilic species and an electrophilic species. It is an extremely useful compound.
• the fused ring tripeptide compound of the present invention is also possible to produce a novel silane-containing fused ring tripeptide compound (hereinafter may be abbreviated as "the fused ring tripeptide compound of the present invention", etc.) using the fused ring dipeptide compound of the present invention. It is.
• a novel silane-containing fused ring tripeptide compound and its manufacturing method which is one of the gist of the present invention, will also be explained ([VI. The fused ring tripeptide compound of the present invention and its manufacturing method]).
• a novel method for producing a polypeptide using the fused ring tripeptide compound of the present invention will also be explained ([VII. Method for producing a polypeptide using the fused ring tripeptide compound of the present invention]).
• amino acid means a compound having a carboxyl group and an amino group.
• the type of amino acid is not particularly limited.
• it may be a D form, an L form, or a racemic form.
• it may be any of ⁇ -amino acids, ⁇ -amino acids, ⁇ -amino acids, ⁇ -amino acids, ⁇ -amino acids, etc.
• amino acids include, but are not limited to, natural amino acids that constitute proteins, and specific examples include valine, leucine, isoleucine, alanine, arginine, glutamine, lysine, aspartic acid, and glutamic acid. , proline, cysteine, threonine, methionine, histidine, phenylalanine, tyrosine, tryptophan, asparagine, glycine, serine and the like.
• peptide refers to a compound in which multiple amino acids are linked via peptide bonds.
• the plurality of amino acid units constituting the peptide may be of the same type, or may be two or more different types of amino acid units.
• the number of amino acids constituting the peptide is not particularly limited as long as it is 2 or more. Examples include 2 (also referred to as a "dipeptide"), 3 (also referred to as a "tripeptide"), 4 (also referred to as a "tetrapeptide”), 5 (also referred to as a "pentapeptide"), 6, 7, 8, 9 , 10, 15, 20, 30, 40, 50, 100, or more.
• a peptide larger than a tripeptide is sometimes referred to as a "polypeptide.”
• an "amino group” refers to a group obtained by removing hydrogen from ammonia, a primary amine, or a secondary amine, each having the formula -NH 2 , -NRH, or -NRR' (where R and R' each means a substituent.) means a functional group represented by.
• hydrocarbon groups may be aliphatic or aromatic.
• the aliphatic hydrocarbon group may be chain or cyclic.
• the chain hydrocarbon group may be linear or branched.
• the cyclic hydrocarbon group may be monocyclic, bridged cyclic, or spirocyclic.
• the hydrocarbon group may be saturated or unsaturated, in other words may contain one or more carbon-carbon double bonds and/or triple bonds. That is, the term "hydrocarbon group” includes alkyl groups, alkenyl groups, alkynyl groups, cycloalkyl groups, cycloalkenyl groups, cycloalkynyl groups, aryl groups, and the like.
• one or more hydrogen atoms in the hydrocarbon group may be substituted with any substituent, and one or more carbon atoms in the hydrocarbon group may be substituted with any substituent depending on the valence. It may be replaced with any heteroatom.
• hydrocarbon oxy group means a group in which the above-defined hydrocarbon group is connected to one bond of an oxy group (-O-).
• the heterocyclic group may be saturated or unsaturated, in other words, it may contain one or more carbon-carbon double bonds and/or triple bonds. Further, the heterocyclic group may be monocyclic, bridged cyclic, or spirocyclic. Furthermore, the heteroatoms contained in the heterocyclic atoms of the heterocyclic group are not limited, but examples include nitrogen, oxygen, sulfur, phosphorus, silicon, and the like.
• amino acids and their residues may be represented by three-letter abbreviations that are well known to those skilled in the art.
• the three letter abbreviations of the main amino acids used in this disclosure are shown in the table below.
• ⁇ -homoamino acids and their residues may be represented by adding “Ho” in front of the three-letter abbreviation of the corresponding ⁇ -amino acid.
• fused ring dipeptide compound of the present invention One aspect of the present invention relates to a novel silane-containing fused ring dipeptide compound represented by the following formula (A) (hereinafter may be abbreviated as "the fused ring dipeptide compound of the present invention", etc.) as appropriate.
• R 11 , R 12 , R 13 , R 21 , and/or R 22 is a monovalent aliphatic hydrocarbon group, aromatic, which may have one or more substituents.
• a linking group is interposed between the aliphatic hydrocarbon group, aromatic hydrocarbon group, or heterocyclic group and the carbon atom to which it is bonded. You may do so.
• Such linking groups are not limited to, but are each independently selected from, for example, the structures shown below (in the following chemical formula, each A independently has one or more substituents). represents a monovalent aliphatic hydrocarbon group, aromatic hydrocarbon group, or heterocyclic group, which may be ).
• R 11 , R 12 , R 13 , R 21 , and/or R 22 is an aromatic hydrocarbon group (which may have one or more substituents), such
• the number of carbon atoms of the aromatic hydrocarbon group is not particularly limited, but the upper limit is, for example, 20 or less, 15 or less, 10 or less, 8 or less, or 6 or less. etc.
• the lower limit varies depending on the type of aromatic hydrocarbon group, but is usually 4 or more, for example 5 or more, or 6 or more. Specific examples of the number of atoms are, for example, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.
• R a1 and R a2 each independently represent a monovalent aliphatic hydrocarbon group or an aromatic hydrocarbon group which may have one or more substituents. When these groups have a substituent, the type thereof is arbitrarily selected from those detailed above. The number of substituents is also not limited, and is, for example, 5, 4, 3, 2, 1, or 0.
• R a1 and/or R a2 is an aliphatic hydrocarbon group (which may have one or more substituents), the substituent of the aliphatic hydrocarbon group (if it has a substituent)
• the number of carbon atoms is not particularly limited, but the upper limit is, for example, 20 or less, 15 or less, 10 or less, 8 or less, or 6 or less.
• the lower limit varies depending on the type of aliphatic hydrocarbon group, but is 1 or more for alkyl groups, 2 or more for alkenyl or alkynyl groups, and 3 or more for cycloalkyl groups, such as 4 or more, or 5 or more. It is.
• Specific examples of the number of atoms include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. It is.
• the side chains of these ⁇ -amino acids may contain one or more of the above-mentioned substituents (for example, halogen, etc.) and/or one or more of the below-mentioned protecting groups (carboxyl group protecting group and/or amino (protecting group), such as t-butyl substituted asparagine, t-butyl substituted glutamine, t-butyl substituted serine, t-butyl substituted threonine, t-butyl substituted tryptophan, t-butyl substituted lysine, Boc Substituted asparagine, Boc substituted glutamine, Boc substituted serine, Boc substituted threonine, Boc substituted tryptophan, Boc substituted lysine, t-butyl substituted aspartic acid, t-butyl substituted glutamic acid, trityl substituted asparagine, trityl substituted glutamine, trityl substituted substituted
• the N-terminal amino group (HNR 13 -) of the first amino acid of formula (R1) and the C-terminal carboxyl group (-COOH) of the second amino acid of formula (R2) are both unprotected.
• the method for producing a fused ring dipeptide compound of the present invention can produce a silane-containing fused ring dipeptide having a desired structure even when unprotected amino acids are used as both the first amino acid which is an electrophilic species and the second amino acid which is a nucleophilic species.
• a major advantage is that compounds can be specifically synthesized.
• n b represents an integer of 1 or 2.
• each of the aforementioned groups of Z b has a substituent
• the type thereof is as described above, but among them, an alkyl group (for example, a straight or branched chain having 1 to 10 carbon atoms) ), alkoxy group (-OR), amino group (-NH 2 ), alkylamino group (-NHR), dialkylamino group (-NR 2 : two
• the alkyl groups R may be the same or different.
• thioalkyl groups (-SR) and groups in which these groups are substituted with one or more halogen atoms (for example, bromine or chlorine atoms) are preferred.
• Specific examples of the number of substituents are, for example, 10, 9, 8, 7, 6, 5, 4, 3, 2, 1, or 0. When the number of substituents is two or more, these may be the same or different.
• step (ii) the second silane compound of formula (S2) forms a silyl ester with the second unprotected amino acid, which is a nucleophile, and protects the terminal carboxyl group of the second amino acid. do.
• step (iii) by mixing both reactants, the terminal carboxyl group of the first amino acid is differentiated from the first amino acid, which is an electrophilic species, and the second amino acid, which is an electrophilic species. It is presumed that this makes it possible to specifically react with the terminal amino group of the second amino acid.
• TMS-OTf trimethylsilyl trifluoromethanesulfonate
• dimethylsilylimidazole dimethylsilyl(2-methyl)imidazole, 1-(trimethylsilyl)imidazole (TMSIM), dimethylethylsilylimidazole (DMESI), dimethylisopropylsilylimidazole (DMIPSI) ), 1-(tert-butyldimethylsilyl)imidazole (TBSIM), 1-(trimethylsilyl)triazole, 1-(tert-butyldimethylsilyl)triazole, N-methyl-Ntrimethylsilyltrifluoroacetamide (MSTFA), N, O-bis(trimethylsilyl)trifluoroacetamide (BSTFA), N,O-bis(trimethylsilyl)acetamide (BSA), N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide (MTB
• step (iii) may include a step of mixing the reaction system with a fourth silane compound.
• the fourth silane compound is not essential, carrying out step (iii) in the presence of the fourth silane compound in the reaction system provides various advantages such as improved reaction efficiency and reaction specificity. There may be cases where Without being bound by theory, even if Si-OH is generated from the first silane compound and/or the second silane compound after the peptide bond is formed in step (iii), the fourth silane It is presumed that Si-OH can be blocked as Si-O-Si due to the coexistence of the compound.
• TMS-OTf trimethylsilyl trifluoromethanesulfonate
• dimethylsilylimidazole dimethylsilyl(2-methyl)imidazole, 1-(trimethylsilyl)imidazole (TMSIM), dimethylethylsilylimidazole (DMESI), dimethylisopropylsilylimidazole (DMIPSI) ), 1-(tert-butyldimethylsilyl)imidazole (TBSIM), 1-(trimethylsilyl)triazole, 1-(tert-butyldimethylsilyl)triazole, N-methyl-Ntrimethylsilyltrifluoroacetamide (MSTFA), N, O-bis(trimethylsilyl)trifluoroacetamide (BSTFA), N,O-bis(trimethylsilyl)acetamide (BSA), N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide (MTB
• a Lewis acid catalyst may be present in the reaction system.
• a Lewis acid catalyst By carrying out the reaction in the presence of a Lewis acid catalyst in the reaction system, various advantages such as improved reaction yield and stereoselectivity may be obtained.
• a Lewis acid catalyst it may be necessary to separate and remove the Lewis acid catalyst from the reaction product. Therefore, it is preferable to appropriately decide whether or not to use a Lewis acid catalyst, taking into consideration the purpose of using the production method of the present invention.
• the type thereof is not limited, but it is preferably a metal compound that functions as a Lewis acid.
• the metal elements constituting the metal compound include various metals belonging to Groups 2 to 15 of the Periodic Table of Elements. Specific examples of the metal elements include boron, magnesium, aluminum, gallium, indium, silicon, calcium, lead, bismuth, mercury, transition metals, lanthanoid elements, and the like.
• transition metals include scandium, titanium, vanadium, chromium, manganese, iron, cobalt, nickel, copper, zinc, yttrium, zirconium, niobium, molybdenum, technetium, ruthenium, rhodium, palladium, tin, silver, cadmium, Examples include hafnium, tantalum, tungsten, rhenium, osmium, iridium, platinum, gold, and thallium.
• lanthanoid elements include lanthanum, cerium, neodymium, samarium, europium, gadolinium, holmium, erbium, thulium, ytterbium, and the like.
• titanium, zirconium, hafnium, tantalum, niobium, boron, vanadium, tungsten, neodymium, iron, lead, and cobalt are preferred from the viewpoint of producing amide compounds with high stereoselectivity and exhibiting an excellent reaction promotion effect.
• metal elements contained in the metal compound may be one or two or more. When the metal compound contains two or more metal elements, these may be of the same type, or may be two or more different metal elements.
• the ligands constituting the metal compound are appropriately selected depending on the type of metal.
• Specific examples of the ligand include substituted or unsubstituted linear or branched chains having 1 to 10 carbon atoms, such as methoxy, ethoxy, propoxy, butoxy, trifluoroethoxy, and trichloroethoxy groups.
• alkoxy groups halogen atoms such as fluorine, chlorine, bromine, and iodine; allyloxy groups having 1 to 10 carbon atoms; acetylacetonate groups (acac), acetoxy groups (AcO), trifluoromethanesulfonate groups ( TfO); substituted or unsubstituted linear or branched alkyl group having 1 to 10 carbon atoms; phenyl group, oxygen atom, sulfur atom, group -SR (where R is a substituent, Examples include substituted or unsubstituted hydrocarbon groups having about 1 to 20 carbon atoms.), group -NRR' (where R and R' are each independently a hydrogen atom or a substituent). Examples of substituents include substituted or unsubstituted hydrocarbon groups having about 1 to 20 carbon atoms), cyclopentadienyl (Cp) groups, and the like.
• substituents include substituted or unsubstituted hydrocarbon groups having
• a titanium compound a zirconium compound, a hafnium compound, a tantalum compound, or a niobium compound is preferable. Specific examples of each are listed below. Incidentally, any one of these may be used alone, but two or more may be used in combination in any combination and ratio.
• a specific example of a titanium compound is TiX 1 4 (However, each of the four X 1s is independently a ligand as exemplified above. The four X 1s may be the same ligand or may be different from each other. ) may be mentioned.
• X 1 is an alkoxy group, preferably a straight chain or branched alkoxy group having 1 to 10 carbon atoms, especially a straight chain or branched alkoxy group having 1 to 5 carbon atoms, and more preferably a straight chain or branched alkoxy group having 1 to 5 carbon atoms. -4 linear or branched alkoxy groups, etc.
• X 1 is an allyloxy group, preferably an allyloxy group having 1 to 20 carbon atoms, particularly an allyloxy group having 1 to 15 carbon atoms, and more preferably an allyloxy group having 1 to 10 carbon atoms.
• These ligands may further have a substituent.
• X 1 is a halogen atom, preferred examples include a chlorine atom and a bromine atom.
• X 2 is an allyloxy group, preferably an allyloxy group having 1 to 20 carbon atoms, particularly an allyloxy group having 1 to 15 carbon atoms, and more preferably an allyloxy group having 1 to 10 carbon atoms.
• These ligands may further have a substituent.
• X 2 is a halogen atom, preferred examples include a chlorine atom and a bromine atom.
• X 3 is an allyloxy group, preferably an allyloxy group having 1 to 20 carbon atoms, particularly an allyloxy group having 1 to 15 carbon atoms, and more preferably an allyloxy group having 1 to 10 carbon atoms.
• These ligands may further have a substituent.
• X 3 is a halogen atom, preferred examples include a chlorine atom and a bromine atom. Among these, for example, HfCp 2 Cl 2 , HfCpCl 3 , HfCl 4 and the like are preferred.
• a specific example of a tantalum compound is TaX 4 5 (However, each of the five X 4s is independently a ligand as exemplified above. The five X 4s may be the same ligand or may be different from each other. ) is exemplified.
• X 4 is an alkoxy group, it is preferably a straight or branched alkoxy group having 1 to 10 carbon atoms, especially a straight or branched alkoxy group having 1 to 5 carbon atoms, and more preferably a straight or branched alkoxy group having 1 to 5 carbon atoms. -3 linear or branched alkoxy groups, etc.
• tantalum alkoxide compounds for example, compounds in which X 4 is an alkoxy group
• tantalum alkoxide compounds such as Ta(OMe) 5 , Ta(OEt) 5 , Ta(OBu) 5 , Ta(NMe 2 ) 5 , Ta(acac)(OEt) 4 , TaCl 5 , TaCl 4 (THF), TaBr 5 and the like are preferred.
• Compounds in which X 4 is oxygen, ie Ta 2 O 5 can also be used.
• niobium compound is NbX 5 5 (however, each of the five X 5s is independently a ligand as exemplified above. The five X 5s may be the same ligand or may be different from each other).
• examples include niobium compounds represented by: When X 5 is an alkoxy group, preferably a straight chain or branched alkoxy group having 1 to 10 carbon atoms, especially a straight chain or branched alkoxy group having 1 to 5 carbon atoms, and more preferably a straight chain or branched alkoxy group having 1 to 5 carbon atoms. -3 linear or branched alkoxy groups, etc.
• X 5 is an allyloxy group, preferably an allyloxy group having 1 to 20 carbon atoms, particularly an allyloxy group having 1 to 15 carbon atoms, and more preferably an allyloxy group having 1 to 10 carbon atoms.
• These ligands may further have a substituent.
• X 5 is a halogen atom, preferred examples include a chlorine atom and a bromine atom.
• niobium alkoxide compounds for example, compounds in which X 5 is an alkoxy group
• NbCl 4 (THF) NbCl 5 , Nb(OMe) 5 , Nb(OEt) 5 and the like are preferable.
• Compounds in which X 5 is oxygen, ie Nb 2 O 5 can also be used.
• the Lewis acid catalyst may be supported on a carrier.
• the carrier supporting the Lewis acid catalyst is not particularly limited, and any known carrier can be used. Furthermore, known methods can be employed as a method for supporting the Lewis acid catalyst on a carrier.
• a base may be present in the reaction system from the viewpoint of increasing reaction efficiency.
• the type of base is not limited, and any known base known to improve reaction efficiency can be used.
• examples of such bases include those having 1 to 1 carbon atoms, such as tetrabutylammonium fluoride (TBAF), triethylamine (Et 3 N), diisopropylamine (i-Pr 2 NH), diisopropylethylamine (i-Pr 2 EtN), etc.
• examples include amines having 1 to 4 linear or branched alkyl groups of 10. Any one of these may be used alone, or two or more may be used in combination in any combination and ratio.
• ⁇ Other ingredients In the method for producing a fused ring dipeptide compound of the present invention, other components may be present in the reaction system. Examples of such other components include, but are not limited to, iodine, trimethylsilyl chloride, trimethylsilyl bromide, trimethylsilyl iodide, and the like. Any one of these may be used alone, or two or more may be used in combination in any combination and ratio.
• the reaction may be carried out in a solvent.
• the solvent is not particularly limited, and examples thereof include aqueous solvents and organic solvents.
• Organic solvents include, but are not limited to, aromatic hydrocarbons such as toluene and xylene, pentane, petroleum ether, tetrahydrofuran (THF), 1-methyltetrahydrofuran (1-MeTHF), diisopropyl ether (i-Pr 2 O), diethyl ether (Et 2 O), ethers such as cyclopentyl methyl ether (CPME), nitrogen-based organic solvents such as acetonitrile (MeCN), chlorine-based organic solvents such as dichloromethane (DCM), ethyl acetate (AcOEt) and organic acids such as acetic acid. These solvents may be used alone or in combination of two or more.
• step (i) the first amino acid of formula (R1) among the substrate compounds is brought into contact with the first silane compound of formula (S1) to react;
• step (ii) the other substrate compound, the amino acid of formula (R2), is brought into contact with the second silane compound of formula (S2) to react.
• step (iii) the reactant of step (i) and the reactant of step (ii) are mixed and further reacted to obtain the fused ring dipeptide compound of formula (A).
• the fused ring dipeptide compound of the present invention is formed by the first amino acid of formula (R1) which is an electrophilic species and the second amino acid of formula (R2) which is a nucleophilic species.
• the reaction mechanism is not bound by theory, the present inventors speculate as follows. That is, in step (i), the first silane compound of formula (S1) forms a five-membered ring with the first unprotected amino acid, which is an electrophilic species, and protects the terminal amino group of the first amino acid.
• step (ii) the second silane compound of formula (S2) forms a silyl ester with the second unprotected amino acid, which is a nucleophile, and protects the terminal carboxyl group of the second amino acid. do.
• step (iii) by mixing both reactants, the terminal carboxyl group of the first amino acid is differentiated from the first amino acid, which is an electrophilic species, and the second amino acid, which is an electrophilic species. It is presumed that this makes it possible to specifically react with the terminal amino group of the second amino acid.
• the timing of adding other components such as the optionally used third and/or fourth silane compound, Lewis acid catalyst, and base to the reaction system is not particularly limited, and they may be added at any timing.
• the third silane compound it is preferably added to the system at the start of step (i).
• the fourth silane compound it is preferably added to the system at the start of step (iii).
• a Lewis acid catalyst it is preferably added to the system at the start of step (ii).
• a base it is preferably added to the system at the start of step (i).
• the components may be mixed in the solvent and brought into contact with each other.
• the amounts of each component used are not limited, but are preferably as follows.
• the amount of the first silane compound of formula (S1) to be used is not particularly limited as long as it does not interfere with the reaction, but for example, the amount of the first silane compound of formula (S1) per mole of the amino acid of formula (R1) For example, 0.1 mol or more, or 0.2 mol or more, or 0.3 mol or more, or 0.4 mol or more, or 0.5 mol or more, and also, for example, 20 mol or less, or 15 mol or less, or 10 It can be used in a range of mol or less, 8 mol or less, 6 mol or less, 4 mol or less, or 2 mol or less.
• the total amount of the two or more types of first silane compounds of formula (S1) may satisfy the above range. .
• the amount of the second silane compound of formula (S2) to be used is not particularly limited as long as it does not interfere with the reaction, but for example, the amount of the second silane compound of formula (S2) per mole of the amino acid of formula (R2) For example, 0.2 mol or more, or 0.4 mol or more, or 0.6 mol or more, or 0.8 mol or more, or 1.0 mol or more, and, for example, 40 mol or less, or 30 mol or less, or 20 It can be used in a range of mol or less, 15 mol or less, 10 mol or less, 6 mol or less, or 4 mol or less.
• the total amount of the two or more types of second silane compounds of formula (S2) may satisfy the above range. .
• the amount used is not particularly limited as long as it does not interfere with the reaction. mol or more, or 0.2 mol or more, or 0.3 mol or more, or 0.4 mol or more, or 0.5 mol or more, and also, for example, 20 mol or less, or 15 mol or less, or 10 mol or less, or 8 It can be used in a range of mol or less, 6 mol or less, 4 mol or less, or 2 mol or less. Note that when two or more types of third silane compounds are used together, the total amount of the two or more types of third silane compounds may satisfy the above range.
• the amount used is not particularly limited as long as it does not interfere with the reaction, but for example, 0.2 mol or more, or 0.4 mol or more, or 0.6 mol or more, or 0.8 mol or more, or 1.0 mol or more, and also, for example, 40 mol or less, or 30 mol or less, or 20 mol or less, or 15 It can be used in a range of mol or less, 10 mol or less, 6 mol or less, or 4 mol or less.
• the total amount of the two or more types of fourth silane compounds may satisfy the above range.
• the amount used is not particularly limited, but for example, 0.2 mol or more, or 0.4 mol or more, or 0.6 mol or more, per 1 mol of the amino acid of formula (R1), or 0.8 mole or more, or 1.0 mole or more, and also, for example, 40 mole or less, or 30 mole or less, or 20 mole or less, or 15 mole or less, or 10 mole or less, or 6 mole or less, or 4 mole or less It can be used within the range of.
• the amount used is not particularly limited, but when the amount of the amino acid of formula (R1) is 100 mol%, it is usually 0.1 mol% or more, for example 0.2 mol% or more, or 0. 3 mol% or more, and usually 30 mol% or less, for example 20 mol% or less, or 15 mol% or less of Lewis acid catalyst can be used.
• reaction conditions in the method for producing a fused ring dipeptide compound of the present invention are not limited as long as the reaction proceeds, but examples for each reaction procedure are as follows.
• reaction conditions for bringing the amino acid of formula (R1) into contact with the first silane compound of formula (S1) to react are not limited as long as the reaction proceeds, but for example, as follows. It is.
• reaction temperature in step (i) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• the reaction pressure in step (i) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (i) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (i) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• reaction conditions for bringing the amino acid of formula (R2) into contact with the second silane compound of formula (S2) to react are not limited as long as the reaction proceeds, but for example, as follows. It is.
• reaction temperature in step (ii) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• reaction pressure in step (ii) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (ii) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (ii) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• reaction conditions for bringing the reactant of step (i) and the reactant of step (ii) into contact and reacting are not limited as long as the reaction proceeds, but for example, the following conditions may be used. be.
• reaction pressure in step (iii) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (iii) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• steps (i), (ii), and (iii) may each be carried out in a sequential method (batch method) or in a continuous method (flow method). Details of specific sequential (batch) and continuous (flow) method implementation procedures are known in the art. Alternatively, step (i) and step (iii) may be performed continuously in one pod by adding the reactant of step (ii) to the reaction system of step (i).
• the fused ring dipeptide compound of the present invention obtained by the above production method may be further subjected to various post-treatments.
• the produced fused ring dipeptide compound of the present invention can be isolated and purified according to conventional methods such as column chromatography and recrystallization.
• the produced fused ring dipeptide compound of the present invention may be used for producing a polypeptide, either directly or after isolation and purification, by subjecting it to the method for producing a polypeptide of the present invention described below.
• the fused ring dipeptide compound of the present invention can be used in various reactions, it is particularly suitable for use in the production of polypeptides.
• the method for producing a polypeptide using the fused ring dipeptide compound of the present invention includes two types of embodiments (hereinafter, these embodiments will be referred to as "the first method for producing a polypeptide of the present invention” and “the method of producing the first polypeptide of the present invention”). ).
• the method for producing a polypeptide using the fused ring dipeptide compound of the present invention is not limited to these two embodiments.
• the first method for producing a polypeptide of the present invention is a method in which one molecule of the fused ring dipeptide compound of the present invention is used for producing one molecule of a polypeptide compound, the method comprising: a fused ring dipeptide compound of the formula (A); By reacting a protected amino acid or a protected peptide compound represented by the following formula (R3) with an amino acid ester or a peptide ester compound represented by the following formula (R4), a polypeptide represented by the following formula (P1) is produced.
• a method comprising obtaining a compound.
• ⁇ Silane-containing fused ring dipeptide compound (substrate compound) The amino acid used as a substrate compound in the first method for producing a polypeptide of the present invention is a silane-containing condensed ring dipeptide compound represented by the above formula (A) (the condensed ring dipeptide compound of the present invention). The details are as described above.
• ⁇ Protected amino acids ⁇ Peptides and amino acids ⁇ Peptide esters are compounds represented by the following formulas (R3) and (R4), respectively. be.
• R 31 , R 32 , R 41 , and R 42 each independently represent a hydrogen atom, a halogen atom, a hydroxyl group, a carboxyl group, a nitro group, a cyano group, or a thiol group, Alternatively, it represents an amino group, a monovalent aliphatic hydrocarbon group, a monovalent aromatic hydrocarbon group, or a monovalent heterocyclic group, which may have one or more substituents. In addition, when these groups have a substituent, the type is as described above. Specific examples of the number of substituents are, for example, 5, 4, 3, 2, 1, or 0.
• R 31 , R 32 , R 41 , and/or R 42 is a monovalent aliphatic hydrocarbon group which may have one or more substituents. , an aromatic hydrocarbon group, or a heterocyclic group, there is a linking group between the aliphatic hydrocarbon group, aromatic hydrocarbon group, or heterocyclic group and the carbon atom to which it is bonded. may be present.
• Such linking groups are not limited to, but are each independently selected from, for example, the structures shown below (in the following chemical formula, each A independently has one or more substituents). represents a monovalent aliphatic hydrocarbon group, aromatic hydrocarbon group, or heterocyclic group, which may be ).
• R 31 , R 32 , R 41 , and/or R 42 is an aliphatic hydrocarbon group
• the aliphatic hydrocarbon group (if it has a substituent)
• the number of carbon atoms (including the substituents) is not particularly limited, but the upper limit is, for example, 20 or less, 15 or less, 10 or less, 8 or less, or 6 or less.
• the lower limit varies depending on the type of aliphatic hydrocarbon group, but is 1 or more for alkyl groups, 2 or more for alkenyl or alkynyl groups, and 3 or more for cycloalkyl groups, such as 4 or more, or 5 or more. It is.
• Specific examples of the number of atoms include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. It is.
• R 31 , R 32 , R 41 , and/or R 42 is an aromatic hydrocarbon group
• the aromatic hydrocarbon group if it has a substituent
• the number of carbon atoms (including substituents) is not particularly limited, but the upper limit is, for example, 20 or less, 15 or less, 10 or less, 8 or less, or 6 or less.
• the lower limit varies depending on the type of aromatic hydrocarbon group, but is usually 4 or more, for example 5 or more, or 6 or more. Specific examples of the number of atoms are, for example, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.
• R 31 , R 32 , R 41 , and/or R 42 when R 31 , R 32 , R 41 , and/or R 42 is a heterocyclic group, the heterocyclic group (if any, the substituent)
• the total number of carbon atoms (including groups) and heteroatoms is not particularly limited, but the upper limit is, for example, 20 or less, 15 or less, 10 or less, 8 or less, or 6 or less.
• the lower limit varies depending on the type of heterocyclic structure, but is usually 3 or more, for example 4 or more, or 5 or more. Specific examples of the number of atoms are, for example, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.
• R 31 , R 32 , R 41 , and R 42 in formula (R3) and formula (R4) each independently represent a hydrogen atom, a hydroxyl group, a thiol group, a carboxyl group, a nitro group, a cyano group, or a halogen atom, Or, an amino group, an alkyl group, an alkenyl group, a cycloalkyl group, an alkoxy group, an aryl group, an aryloxy group, an acyl group, a heterocyclic group, or a heterocyclic group, which may have one or more substituents.
• it is an oxy group or the like.
• R 31 , R 32 , R 41 , and R 42 in formula (R3) and formula (R4) include, but are not limited to, the following.
• ⁇ Hydrogen atom hydroxyl group, thiol group, carboxyl group, nitro group, cyano group
• ⁇ Halogen atoms such as fluorine atoms, chlorine atoms, bromine atoms, and iodine atoms
• R 33 and R 43 are each independently a hydrogen atom, a carboxyl group, a hydroxyl group, or a monovalent group which may have one or more substituents. represents a hydrocarbon group or a heterocyclic group.
• substituents when a substituent is present, the type thereof is as described above. Specific examples of the number of substituents are, for example, 5, 4, 3, 2, 1, or 0.
• R 33 and/or R 43 may have one or more substituents, a monovalent aliphatic hydrocarbon group, an aromatic hydrocarbon group, Or, in the case of a heterocyclic group, a linking group may be interposed between the aliphatic hydrocarbon group, aromatic hydrocarbon group, or heterocyclic group and the nitrogen atom to which it is bonded.
• Such linking groups are not limited to, but are each independently selected from, for example, the structures shown below (in the following chemical formula, each A independently has one or more substituents). represents a monovalent aliphatic hydrocarbon group, aromatic hydrocarbon group, or heterocyclic group, which may be ).
• R 33 and/or R 43 when R 33 and/or R 43 is an aliphatic hydrocarbon group, the aliphatic hydrocarbon group (including the substituent if it has a substituent)
• the number of carbon atoms is not particularly limited, but the upper limit is, for example, 20 or less, 15 or less, 10 or less, 8 or less, or 6 or less.
• the lower limit varies depending on the type of aliphatic hydrocarbon group, but is 1 or more for alkyl groups, 2 or more for alkenyl or alkynyl groups, and 3 or more for cycloalkyl groups, such as 4 or more, or 5 or more. It is.
• Specific examples of the number of atoms include, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. It is.
• R 33 and/or R 43 is an aromatic hydrocarbon group
• the aromatic hydrocarbon group (including the substituent if it has a substituent)
• the number of carbon atoms is not particularly limited, but the upper limit is, for example, 20 or less, 15 or less, 10 or less, 8 or less, or 6 or less.
• the lower limit varies depending on the type of aromatic hydrocarbon group, but is usually 4 or more, for example 5 or more, or 6 or more. Specific examples of the number of atoms are, for example, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.
• R 33 and/or R 43 when R 33 and/or R 43 is a heterocyclic group, the carbon atom of such heterocyclic group (including the substituent if it has a substituent)
• the total number of heteroatoms is not particularly limited, but the upper limit is, for example, 20 or less, 15 or less, 10 or less, 8 or less, or 6 or less.
• the lower limit varies depending on the type of heterocyclic structure, but is usually 3 or more, for example 4 or more, or 5 or more. Specific examples of the number of atoms are 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20, for example.
• R 33 and/or R 43 in formula (R3) and formula (R4) may each independently have a hydrogen atom, a hydroxyl group, or a carboxyl group, or one or more substituents, Preferably, it is an alkyl group, an alkenyl group, a cycloalkyl group, an alkoxy group, an aryl group, an aryloxy group, an acyl group, a heterocyclic group, a heterocyclic oxy group, or the like.
• R 33 and/or R 43 in formula (R3) and formula (R4) include, but are not limited to, the following.
• ⁇ Hydrogen atom hydroxyl group, carboxyl group
• R 31 and R 33 are bonded to each other and have one or more substituents together with the carbon atom to which R 31 is bonded and the nitrogen atom to which R 33 is bonded. It may also form a good heterocycle.
• R 41 and R 43 are bonded to each other and have one or more substituents together with the carbon atom to which R 41 is bonded and the nitrogen atom to which R 43 is bonded. may form a heterocyclic ring.
• the type thereof is as described above. Specific examples of the number of substituents are, for example, 5, 4, 3, 2, 1, or 0.
• the total number of carbon atoms and heteroatoms in such a heterocyclic group has an upper limit of, for example, 20 or less, 15 or less, 10 or less, 8 or less, or 6 or less. It is.
• the lower limit varies depending on the type of heterocyclic structure, but is usually 3 or more, for example 4 or more, or 5 or more. Specific examples of the number of atoms are, for example, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.
• heterocycles include, but are not limited to, pyrrolinyl, pyrrolyl, 2,3-dihydro-1H-pyrrolyl, piperidinyl, piperazinyl, homopiperazinyl, and morpholino groups.
• thiomorpholino group 1,2,4,6-tetrahydropyridyl group, hexahydropyrimidyl group, hexahydropyridazyl group, 1,2,4,6-tetrahydropyridyl group, 1,2,4,6 -tetrahydropyridazyl group, 3,4-dihydropyridyl group, imidazolyl group, 4,5-dihydro-1H-imidazolyl group, 2,3-dihydro-1H-imidazolyl group, pyrazolyl group, 4,5-dihydro-1H -pyrazolyl group, 2,3-dihydro-1H-pyrazolyl group, oxazolyl group, 4,5-dihydro-1,3-oxazolyl group, 2,3-dihydro-1,3-oxazolyl group, 2,5-dihydro- 1,3-oxazolyl group, thiazolyl group,
• a 31 , A 32 , A 41 , and A 42 are each independently a carbon number 1 to 3 carbon atom that may have one or more substituents. represents a valent aliphatic hydrocarbon group. Specific examples include, but are not limited to, methylene groups, ethylene groups, propylene groups, isopropylene groups, and groups in which these groups are substituted with one or more of the above substituents. Can be mentioned. Specific examples of the number of substituents are, for example, 3, 2, 1, or 0.
• p31, p32, p41, and p42 each independently represent 0 or 1.
• m and n are each independently an integer of 1 or more representing the number of structural units represented by the structure in [ ]. That is, m represents the number of amino acid units in [ ] in formula (R3). When m is 1, the compound of formula (R3) becomes a protected amino acid, and when m is 2 or more, the compound of formula (R3) becomes a protected peptide. Similarly, n represents the number of amino acid units in [ ] in formula (R4). When n is 1, the compound of formula (R4) becomes an amino acid ester, and when n is 2 or more, the compound of formula (R4) becomes a peptide ester.
• m and n are not particularly limited as long as the reaction proceeds, but for example, 100 or less, 80 or less, 60 or less, 50 or less, 40 or less, 30 or less, 20 or less, 15 or less, 12 or less, or 10 or less. It is. Specific examples of m and n are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, etc.
• PG a represents a protecting group for an amino group.
• the protecting group PG a for an amino group is not particularly limited as long as it can protect the amino group from reacting during a given reaction and can be deprotected and converted to an amino group after the reaction. Not done. Details of such a protecting group for an amino group will be described later.
• PG b represents a carboxyl group protecting group.
• the carboxyl group protecting group PGb is particularly limited as long as it can protect the carboxyl group from reacting during a given reaction and can be deprotected and converted into a carboxyl group after the reaction. Not done.
• the details of the carboxyl group protecting group are as described above.
• the amino group protecting group PG a used in each production method of the present invention (the method for producing a fused ring dipeptide compound of the present invention, and the method for producing the first and second polypeptides of the present invention) is a known
• a wide variety of types are known. Examples include monovalent aliphatic hydrocarbon groups or aromatic hydrocarbon groups which may have one or more substituents, or monovalent aliphatic hydrocarbon groups or aromatic hydrocarbon groups which may have one or more substituents. valent heterocyclic groups and the like. Among these, monovalent aliphatic hydrocarbon groups or aromatic hydrocarbon groups which may have one or more substituents are preferred.
• linking group may be present between such aliphatic hydrocarbon group, aromatic hydrocarbon group, or heterocyclic group and the nitrogen atom of the amino group it protects (the nitrogen atom to which PG a is bonded in formula (R3)).
• Such linking groups are not limited to, but are each independently selected from, for example, the linking groups shown below (in the following chemical formula, each A independently has one or more substituents). Represents a monovalent aliphatic hydrocarbon group, aromatic hydrocarbon group, or heterocyclic group that may be ).
• the number of carbon atoms in the amino group protecting group PG a is usually 1 or more, or 3 or more, and usually 20 or less, or 15 or less.
• the amino group protecting group PG a is a monovalent aliphatic hydrocarbon group or an aromatic hydrocarbon group, an acyl group, a hydrocarbon oxycarbonyl group, which may have one or more substituents, It is preferably one or more groups selected from the group consisting of a hydrocarbon sulfonyl group, and an amide group.
• amino group protecting group PG a Specific examples of the amino group protecting group PG a are listed below.
• names for the protecting group of an amino group in addition to the name of the functional group bonded to the nitrogen atom of the amino group, there are also names that include the nitrogen atom, and the following names also include both. It is.
• unsubstituted or substituted hydrocarbon groups include alkyl groups such as methyl, ethyl, and propyl; alkenyl groups such as ethenyl, propenyl, and allyl; alkynyl groups such as propargyl; cyclopropyl; cycloalkyl groups such as cyclobutyl, cyclopentyl, and cyclohexyl groups; aryl groups such as phenyl, benzyl, paramethoxybenzyl, tolyl, and triphenylmethyl (troc groups); substituted hydrocarbons such as cyanomethyl Examples include groups. The number of carbon atoms is usually 1 or more, or 3 or more, and usually 20 or less, or 15 or less.
• unsubstituted or substituted acyl groups include benzoyl group (Bz), orthomethoxybenzoyl group, 2,6-dimethoxybenzoyl group, paramethoxybenzoyl group (PMPCO), cinnamoyl group, phthaloyl group (Phth), etc. Can be mentioned.
• unsubstituted or substituted hydrocarbon oxycarbonyl groups include tert-butoxycarbonyl group (Boc), benzyloxycarbonyl group (Cbz or Z), methoxycarbonyl group, ethoxycarbonyl group, 2-trimethylsilylethoxycarbonyl group, 2-phenylethoxycarbonyl group, 1-(1-adamantyl)-1-methylethoxycarbonyl group, 1-(3,5-di-t-butylphenyl)-1-methylethoxycarbonyl group, vinyloxycarbonyl group, allyl Oxycarbonyl group (Alloc), N-hydroxypiperidinyloxycarbonyl group, p-methoxybenzyloxycarbonyl group, p-nitrobenzyloxycarbonyl group, 2-(1,3-dithianyl)methoxycarbonyl, m-nitrophenoxycarbonyl group, 3,5-dimethoxybenzyloxycarbonyl group, o-nitrobenzy
• unsubstituted or substituted hydrocarbon sulfonyl group examples include a methanesulfonyl group (Ms), a toluenesulfonyl group (Ts), a 2- or 4-nitrobenzenesulfonyl group (Ns), and the like.
• Ms methanesulfonyl group
• Ts toluenesulfonyl group
• Ns 2- or 4-nitrobenzenesulfonyl group
• unsubstituted or substituted amide groups include acetamide, o-(benzoyloxymethyl)benzamide, 2-[(t-butyldiphenylsiloxy)methyl]benzamide, 2-toluenesulfonamide, 4-toluenesulfonamide. , 2-nitrobenzenesulfonamide, 4-nitrobenzenesulfonamide, tert-butylsulfinylamide, 4-toluenesulfonamide, 2-(trimethylsilyl)ethanesulfonamide, benzylsulfonamide, and the like.
• a protecting group that can be deprotected by one type of method is also mentioned as an example of the protecting group PG a for an amino group.
• Preferred specific examples of the amino group protecting group PG a include a mesyl group (Ms), a tert-butoxycarbonyl group (Boc), a benzyl group (Bn or Bzl), a benzyloxycarbonyl group (Cbz), and a benzoyl group (Bz).
• Ms mesyl group
• Boc tert-butoxycarbonyl group
• Bn or Bzl a benzyl group
• Cbz benzyloxycarbonyl group
• Bz benzoyl group
• amino group protecting group PG a include mesyl group (Ms), tert-butoxycarbonyl group (Boc), benzyloxycarbonyl group (Cbz), benzyl group (Bn), paramethoxybenzyl group (PMB).
• amino group protecting group PG a examples include mesyl group (Ms), tert-butoxycarbonyl group (Boc), benzyloxycarbonyl group (Cbz), benzyl group (Bn), paramethoxybenzyl group (PMB). ), 2,2,2-trichloroethoxycarbonyl group (Troc), allyloxycarbonyl group (Alloc), paramethoxybenzoyl group (PMPCO), benzoyl group (Bz), cyanomethyl group, cinnamoyl group, and the like.
• Ms mesyl group
• Boc tert-butoxycarbonyl group
• Cbz benzyloxycarbonyl group
• Bn benzyl group
• PMB paramethoxybenzyl group
• Troc 2,2,2-trichloroethoxycarbonyl group
• allyloxycarbonyl group Alloc
• paramethoxybenzoyl group PMPCO
• a condensing agent and/or a racemization inhibitor may be present in the reaction system.
• the method may further include a step of deprotecting the amino group protecting group PG a and/or the carboxyl group protecting group PG b of the polypeptide compound of formula (P1).
• Carboxyl group protecting group examples include monovalent aliphatic hydrocarbon groups, aromatic hydrocarbon groups, and heterocyclic groups that may have one or more substituents.
• a substituent when a substituent is present, the type thereof is as described above. Specific examples of the number of substituents are, for example, 5, 4, 3, 2, 1, or 0.
• the number of carbon atoms in the aliphatic hydrocarbon group is not particularly limited, but the upper limit is, for example, 20 or less, 15 or less, 10 or less, 8 or less, or 6 or less.
• the lower limit varies depending on the type of aliphatic hydrocarbon group, but is 1 or more for alkyl groups, 2 or more for alkenyl or alkynyl groups, and 3 or more for cycloalkyl groups, such as 4 or more, or 5 or more. It is.
• Specific examples of the number of atoms include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. It is.
• the number of carbon atoms in the aromatic hydrocarbon group is not particularly limited, but the upper limit is, for example, 20 or less, 15 or less, 10 or less, 8 or less, or 6 or less.
• the lower limit varies depending on the type of aromatic hydrocarbon group, but is usually 4 or more, for example 5 or more, or 6 or more.
• Specific examples of the number of atoms are, for example, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.
• carboxyl group protecting group PG b examples include, but are not limited to, the following.
• a condensing agent may be present in the system from the viewpoint of increasing the efficiency of the peptide formation reaction.
• the type of condensing agent is not limited, and any known condensing agent known to improve condensation reaction efficiency can be used. Examples of such condensing agents include: Any one of these may be used alone, or two or more may be used in combination in any combination and ratio.
• Carbodiimide condensing agent 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide (wsc, edc), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (wscHCl, edcHCl), N , N'-dicyclohexylcarbodiimide (DCC), N,N'-diisopropylcarbodiimide (DIC), and the like.
• ⁇ Phosphonium condensing agent 1H-benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP), 1H-benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate (PyBOP), (7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate (PyAOP), chlorotripyrrolidinophosphonium hexafluorophosphate (PyCloP), bromotris(dimethylamino)phosphonium Hexafluorophosphate (Brop), 3-(diethoxyphosphoryloxy)-1,2,3-benzotriazin-4(3H)-one (DEPBT), etc.
• BOP 1H-benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate
• - Imidazole condensing agent N,N'-carbonyldiimidazole (CDI), 1,1'-carbonyldi(1,2,4-triazole) (CDT), etc.
• ⁇ Uronium-based condensing agent O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HBTU), O-(7-azabenzotriazole-1 -yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), O-(N-succinimidyl)-N,N,N',N'-tetramethyluronium tetra Fluoroborate (TSTU) etc.
• HBTU O-(benzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate
• HATU O-(7-azabenzotriazole-1 -yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate
• TSTU O-(N-s
• Triazine condensing agent 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride n-hydrate (DMT-MM), etc.
• a racemization inhibitor may be used in combination from the viewpoint of preventing racemization during the peptide formation reaction.
• the type of racemization inhibitor is not limited either, and any known racemization inhibitor known to prevent racemization during the condensation reaction can be used.
• racemization inhibitors include 1-hydroxybenzotriazole (HOBt), 1-hydroxy-7-azabenzotriazole (HOAtN-hydroxysuccinimide (HOSu), N,N'-disuccinimidyl carbonate). (DSC), etc. Any one of these may be used alone, or two or more may be used in combination in any combination and ratio.
• a base may coexist in the system from the viewpoint of increasing reaction efficiency.
• the type of base is not limited, and any known base known to improve reaction efficiency can be used.
• examples of such bases include those having 1 to 1 carbon atoms, such as tetrabutylammonium fluoride (TBAF), triethylamine (Et 3 N), diisopropylamine (i-Pr 2 NH), diisopropylethylamine (i-Pr 2 EtN), etc.
• examples include amines having 1 to 4 linear or branched alkyl groups of 10, and inorganic bases such as cesium fluoride. Any one of these may be used alone, or two or more may be used in combination in any combination and ratio.
• the substrate compound is the above-mentioned fused ring dipeptide compound of formula (A), the protected amino acid or protected peptide of formula (R3), the amino acid ester or peptide ester of formula (R4), and
• other components include, but are not limited to, conventional catalysts that can be used in amidation reactions, silane compounds, phosphorus compounds, and the like. Any one of these may be used alone, or two or more may be used in combination in any combination and ratio.
• catalysts include various Lewis acid catalysts detailed in the section of the method for producing the fused ring dipeptide compound of the present invention, such as titanium compounds, zirconium compounds, hafnium compounds, tantalum compounds, niobium compounds, etc., and methylaluminum compounds.
• Lewis acid catalysts such as titanium compounds, zirconium compounds, hafnium compounds, tantalum compounds, niobium compounds, etc.
• methylaluminum compounds such as titanium compounds, zirconium compounds, hafnium compounds, tantalum compounds, niobium compounds, etc.
• methylaluminum compounds such as titanium compounds, zirconium compounds, hafnium compounds, tantalum compounds, niobium compounds, etc.
• methylaluminum compounds such as titanium compounds, zirconium compounds, hafnium compounds, tantalum compounds, niobium compounds, etc.
• methylaluminum compounds such as titanium compounds, zirconium compounds, hafnium compounds
• silane compounds include HSi(OCH ( CF3 ) 2 ) 3 , HSi( OCH2CF3 ) 3 , HSi( OCH2CF2CF2H ) 3 , HSi( OCH2CF2CF2CF2CF ) .
• trimethylsilyl trifluoromethanesulfonate TMS-OTf
• TMSIM 1-(trimethylsilyl)imidazole
• DMESI dimethylethylsilylimidazole
• DMIPSI dimethylisopropylsilylimidazole
• TBSIM 1-(tert-butyldimethylsilyl)imidazole
• TBSIM 1-(trimethylsilyl)triazole, 1-(tert-butyldimethylsilyl)triazole, dimethylsilylimidazole, dimethylsilyl (2 -Methyl)imidazole
• TMBS trimethylbromosilane
• TMCS trimethylchlorosilane
• MSTFA N,O-bis(trimethylsilyl)trifluoroacetamide
• phosphorus compounds include phosphine compounds (e.g., trimethylphosphine, triethylphosphine, tripropylphosphine, trimethyloxyphosphine, triethyloxyphosphine, triproxyphosphine, triphenylphosphine, trinaphthylphosphine, triphenyloxyphosphine, Tris(4-methylphenyl)phosphine, tris(4-methoxyphenyl)phosphine, tris(4-fluorophenyl)phosphine, tris(4-methylphenyloxy)phosphine, tris(4-methoxyphenyloxy)phosphine, tris(4-fluorophenyl)phosphine -fluorophenyloxy)phosphine etc.), phosphate compounds (e.g., trimethylphosphine, triethylphosphine, tripropylphosphine,
• a solvent may be used during the reaction.
• the solvent is not particularly limited, and examples thereof include aqueous solvents and organic solvents.
• Organic solvents include, but are not limited to, aromatic hydrocarbons such as toluene and xylene, pentane, petroleum ether, tetrahydrofuran (THF), 1-methyltetrahydrofuran (1-MeTHF), diisopropyl ether (i-Pr 2 O), ethers such as diethyl ether (Et 2 O) and cyclopentyl methyl ether (CPME), nitrogen-based organic solvents such as acetonitrile (MeCN), chlorine-based organic solvents such as dichloromethane (DCM), and ethyl acetate (AcOEt). and organic acids such as acetic acid. These solvents may be used alone or in combination of two or more.
• the ring of the amino acid residue on the left side in the formula of the fused ring dipeptide compound of formula (A) opens, and the protected amino acid or protected peptide of formula (R3) is linked to the N-terminus, and )
• the ring of the amino acid residue on the right side in the formula of the fused ring dipeptide compound is opened, and the protected amino acid or protected peptide of formula (R4) is linked to the C-terminus, and as a result, the polypeptide compound of formula (P1) is formed.
• the order of mixing each substrate compound is not limited as long as the above reaction occurs. Examples include the following two embodiments, but the mixing order of each substrate compound is not limited to these.
• step (i) a protected amino acid or a protected peptide of formula (R3) is added to a fused ring dipeptide compound of formula (A) and reacted, and then, as step (ii), a compound of formula Examples include an embodiment in which the amino acid ester or peptide ester of (R4) is added to the reaction system and reacted.
• step (i) the ring of the amino acid residue on the left side in the formula of the fused ring dipeptide compound of formula (A) is opened, and the protected amino acid or protected peptide of formula (R3) is linked to the N-terminus.
• step (ii) the ring of the amino acid residue on the right side in the formula of the fused ring dipeptide compound of formula (A) is opened, and the protected amino acid or protected peptide of formula (R4) is linked to the C-terminus, As a result, a polypeptide compound of formula (P1) is formed.
• step (i) an amino acid ester or peptide ester of formula (R4) is added to the fused ring dipeptide compound of formula (A) and reacted, and then as step (ii), Examples include an embodiment in which the protected amino acid or protected peptide (R3) is added to the reaction system and reacted.
• step (i) the ring of the amino acid residue on the right side in the formula of the fused ring dipeptide compound of formula (A) is opened, and the protected amino acid or protected peptide of formula (R4) is connected to the C-terminus.
• step (ii) the ring of the amino acid residue on the left side in the formula of the fused ring dipeptide compound of formula (A) is opened, and the protected amino acid or protected peptide of formula (R3) is linked to the N-terminus, As a result, a polypeptide compound of formula (P1) is formed.
• timing of adding other components such as an optional condensing agent and a base to the reaction system is not particularly limited, and they may be added at any timing.
• a condensing agent and/or a base it is preferable to add it to the system at the beginning of step (i) and/or step (ii) in both the first and second embodiments.
• a racemization inhibitor is used in addition to the condensing agent, it is preferably added to the system together with the condensing agent.
• the components may be mixed in the solvent and brought into contact with each other.
• the ratio of the fused ring dipeptide compound of formula (A) to the protected amino acid or protected peptide of formula (R3) is not particularly limited, but the ratio of the fused ring dipeptide compound of formula (A) to the fused ring dipeptide compound of formula (R3) is not particularly limited.
• Protected amino acid or protected peptide for example, 0.1 mol or more, or 0.2 mol or more, or 0.3 mol or more, or 0.4 mol or more, or 0.5 mol or more, and, for example, 20 mol or less, Alternatively, it can be used in a range of 15 mol or less, 10 mol or less, 8 mol or less, 6 mol or less, 4 mol or less, or 2 mol or less.
• the quantitative ratio of the fused ring dipeptide compound of formula (A) to the amino acid ester or peptide ester of formula (R4) is not particularly limited, of amino acid ester or peptide ester, for example, 0.1 mol or more, or 0.2 mol or more, or 0.3 mol or more, or 0.4 mol or more, or 0.5 mol or more, and, for example, 20 mol or less, Alternatively, it can be used in a range of 15 mol or less, 10 mol or less, 8 mol or less, 6 mol or less, 4 mol or less, or 2 mol or less.
• the target production amount of the polypeptide compound of formula (P1) of the present invention to be produced is the amount of the fused ring dipeptide compound of formula (A) as a substrate, the protected amino acid or protected peptide of formula (R3). , and the amino acid ester or peptide ester of formula (R4) must be used in an amount of 1 mole or more, respectively.
• the amount used is not particularly limited, but the amount of the base is, for example, 0.2 mol or more, or 0.4 mol or more, or 0.6 mol or more, per 1 mol of the fused ring dipeptide compound of formula (A). mol or more, or 0.8 mol or more, or 1.0 mol or more, and also, for example, 40 mol or less, or 30 mol or less, or 20 mol or less, or 15 mol or less, or 10 mol or less, or 6 mol or less, or It can be used in a range of 4 moles or less.
• the amount used is not particularly limited, but the amount of the condensing agent is, for example, 0.2 mol or more, 0.4 mol or more, or 0. .6 moles or more, or 0.8 moles or more, or 1.0 moles or more, and also, for example, 40 moles or less, or 30 moles or less, or 20 moles or less, or 15 moles or less, or 10 moles or less, or 6 moles or less , or 4 moles or less.
• the amount used is not particularly limited, but for example, 0.2 mol or more of the racemization inhibitor per 1 mol of the fused ring dipeptide compound of formula (A), or 0.4 mol or more, or 0.6 mol or more, or 0.8 mol or more, or 1.0 mol or more, and also, for example, 40 mol or less, or 30 mol or less, or 20 mol or less, or 15 mol or less, Alternatively, it can be used in a range of 10 mol or less, 6 mol or less, or 4 mol or less.
• reaction conditions in the first peptide production method of the present invention are not limited as long as the reaction proceeds, but examples are as follows for each reaction procedure for each of the first and second aspects. .
• reaction conditions for adding and reacting the fused ring dipeptide compound of formula (A) with the protected amino acid or protected peptide of formula (R3) in step (i) are such that the reaction proceeds.
• examples include the following.
• reaction temperature in step (i) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• the reaction pressure in step (i) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (i) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (i) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• step (ii) the reaction conditions when adding the amino acid ester or peptide ester of formula (R4) to the reaction system and reacting are not limited as long as the reaction proceeds, and are, for example, as follows.
• reaction temperature in step (ii) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• reaction pressure in step (ii) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (ii) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (ii) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• reaction conditions for adding and reacting the fused ring dipeptide compound of formula (A) with the amino acid ester or peptide ester of formula (R4) in step (i) are as follows: Although there are no limitations as far as the process progresses, examples include the following.
• reaction temperature in step (i) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• the reaction pressure in step (i) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (i) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (i) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• step (ii) the reaction conditions when adding the protected amino acid or protected peptide of formula (R3) to the reaction system and reacting are not limited as long as the reaction proceeds, and are, for example, as follows.
• reaction temperature in step (ii) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• reaction pressure in step (ii) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (ii) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (ii) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• step (i) and step (ii) may be performed in a sequential method (batch method) or in a continuous method (flow method). It's okay. Details of specific sequential (batch) and continuous (flow) method implementation procedures are known in the art. Alternatively, step (i) and step (ii) may be performed continuously in one pod.
• ⁇ Polypeptide (target compound) The polypeptide compound that is the target compound finally produced in the first polypeptide production method of the present invention is a compound represented by the following formula (P1).
• R 11 , R 12 , R 13 , R 21 , and R 22 represent the same groups as defined in formula (A) above
• PG a , R 31 , R 32 , R 33 , A 31 , A 32 , p31, p32, and m represent the same groups as defined in the formula (R3)
• PG b , R 41 , R 42 , R 43 , A 41 , A 42 , p41, p42, and n are , represents the same group as defined in formula (R4) above.
• the compound of formula (R3) is a protected dipeptide and the compound of formula (R4) is an amino acid ester (that is, when m is 2 and n is 1)
• the compound of formula (R3) is
• the compound of formula (R4) is a protected amino acid and is a dipeptide ester (that is, when m is 1 and n is 2)
• the compound of formula (R3) is a protected dipeptide and the compound of formula (R4) is a dipeptide ester (that is, when m is 2 and n is 2)
• polypeptide compound of formula (P1) obtained by the above-mentioned production method may be further subjected to various post-treatments.
• post-treatments include isolation and purification of the obtained polypeptide compound of formula (P1), deprotection of the amino group-protecting group PG a and/or the carboxyl group-protecting group PG b , and the like. Such post-processing will be described in detail later.
• the second method for producing a polypeptide of the present invention is a method using two molecules of the fused ring dipeptide compound of the present invention for producing one molecule of polypeptide, and includes at least the following steps (i) to (iii). It's a method.
• (i) A step of adding and reacting a protected amino acid or a protected peptide represented by the following formula (R3) to a silane-containing condensed ring dipeptide compound represented by the following formula (A1).
• R3 silane-containing condensed ring dipeptide compound represented by the following formula (A1).
• a step of adding a silane-containing condensed ring dipeptide compound represented by the following formula (A2) to the reaction product of step (i) and further reacting it.
• a polypeptide compound represented by the following formula (P1) is obtained by adding an amino acid ester or a peptide ester represented by the following formula (R4) to the reaction product of step (ii) and further reacting it.
• the silane-containing condensed ring dipeptide compound used as a substrate compound in the method for producing the second polypeptide of the present invention is a silane-containing condensed ring dipeptide compound represented by the above formula (A) (the condensed ring dipeptide compound of the present invention). ), but differs from the method for producing the first polypeptide of the present invention described above in that two molecules of a silane-containing condensed ring dipeptide compound are used to synthesize one molecule of polypeptide.
• they shall be represented by the following formulas (A1) and (A2), respectively.
• R 111 , R 112 , R 113 , R 121 , and R 122 in formula (A1) and R 211 , R 212 , R 213 , R 221 , and R 222 in formula (A2) each independently represent the formula It represents the same definition as R 11 , R 12 , R 21 , and R 22 in (A).
• R a11 and R a12 in formula (A1) and R a21 and R a22 in formula (A2) each independently represent the same definitions as R a1 and R a2 in formula (A). The details are as described above.
• ⁇ Protected amino acids ⁇ Peptides and amino acids ⁇ Peptide esters (substrate compounds):
• the protected amino acid or protected peptide and the amino acid ester or peptide ester used as the substrate compound in the method for producing the second polypeptide of the present invention are expressed by the formulas described above. (R3) and a compound represented by formula (R4). The details are as described above.
• a condensing agent may be present in the system from the viewpoint of increasing the efficiency of the peptide formation reaction.
• a racemization inhibitor may be used in combination. The details of the condensing agent and the racemization inhibitor are as described above in the description of the first peptide production method of the present invention.
• a base may also be allowed to coexist in the system from the viewpoint of increasing reaction efficiency.
• the details of the base are also as described in detail in the explanation of the first peptide production method of the present invention.
• the substrate compounds are the fused ring dipeptide compounds of formula (A1) and formula (A2), the protected amino acid or protected peptide of formula (R3), and the amino acid ester of formula (R4).
• other components may be present in addition to the peptide ester and optionally used base, condensing agent, and racemization inhibitor. Examples include catalysts, silane compounds, phosphorus compounds, and the like. The details of these other components are also as described in detail in the explanation of the first peptide production method of the present invention.
• reaction may be carried out in a solvent.
• a solvent The details of such a solvent are also as described in detail in the explanation of the first peptide production method of the present invention.
• the N-terminus of the latter amino acid residue is connected to the C-terminus of the latter amino acid residue, and the ring of the amino acid residue on the left side of the fused ring dipeptide compound of formula (A1) opens, and the N-terminus of the latter amino acid residue opens.
• the protected amino acid or protected peptide of formula (R3) is linked, and furthermore, the ring of the amino acid residue on the right side in the formula of the fused ring dipeptide compound of formula (A2) is opened to form the protected amino acid or protected peptide of formula (R4). are linked, resulting in the formation of a polypeptide compound of formula (P2).
• the order of mixing each substrate compound is not limited as long as the above reaction occurs. Examples include the following two embodiments, but the mixing order of each substrate compound is not limited to these.
• step (i) a protected amino acid or a protected peptide of formula (R3) is added to the fused ring dipeptide compound of formula (A1) and reacted, and then as step (ii), a fused ring dipeptide compound of formula (A1) is reacted with ) is added to the reaction system and reacted, and as step (iii), the amino acid ester or peptide ester of formula (R4) is added to the reaction system and reacted.
• step (i) the ring of the amino acid residue on the left side in the formula of the fused ring dipeptide compound of formula (A1) is opened, and the protected amino acid or protected peptide of formula (R3) is connected to the N-terminus.
• step (ii) the ring of the amino acid residue on the right side in the formula of the fused ring dipeptide compound of formula (A1) and the ring of the amino acid residue on the left side of the formula of the fused ring dipeptide compound of formula (A2) are both opened. Then, the N-terminus of the latter amino acid residue is linked to the C-terminus of the former amino acid residue, and in step (iii), the ring of the amino acid residue on the right side of the formula (A2) is Upon opening, a protected amino acid or protected peptide of formula (R4) is linked, resulting in the formation of a polypeptide compound of formula (P2).
• step (i) an amino acid ester or peptide ester of formula (R4) is added to the fused ring dipeptide compound of formula (A2) and reacted, and then as step (ii), the fused ring dipeptide compound of formula (A2) is reacted. ) is added to the reaction system and reacted, and as step (iii), the protected amino acid or protected peptide of formula (R3) is added to the reaction system and reacted.
• step (i) the ring of the amino acid residue on the right side in the formula of the fused ring dipeptide compound of formula (A2) is opened, and the protected amino acid or protected peptide of formula (R4) is linked
• step (ii) the ring of the amino acid residue on the right side in the formula of the fused ring dipeptide compound of formula (A1) and the ring of the amino acid residue on the left side of the formula of the fused ring dipeptide compound of formula (A2) are both opened, and the former
• the N-terminus of the latter amino acid residue is connected to the C-terminus of the amino acid residue
• step (iii) the ring of the left amino acid residue in the fused ring dipeptide compound of formula (A1) is opened, and the ring of the amino acid residue on the left in the formula (A1) is opened.
• a protected amino acid or a protected peptide of formula (R3) is linked to the N-terminus, resulting in the formation of a polypeptide compound of
• timing of adding other components such as an optional condensing agent and a base to the reaction system is not particularly limited, and they may be added at any timing.
• a condensing agent and/or a base in both the first and second embodiments, it is necessary to It is preferable to add.
• a racemization inhibitor is used in addition to the condensing agent, it is preferably added to the system together with the condensing agent.
• the components may be mixed in the solvent and brought into contact with each other.
• the ratio of the fused ring dipeptide compound of formula (A1) to the fused ring dipeptide compound of formula (A2) is not particularly limited, but the ratio of the fused ring dipeptide compound of formula (A2) to 1 mol of the fused ring dipeptide compound of formula (A1) is
• the fused ring dipeptide compound may be, for example, 0.1 mol or more, or 0.2 mol or more, or 0.3 mol or more, or 0.4 mol or more, or 0.5 mol or more, and, for example, 20 mol or less, or 15 mol or more. It can be used in a range of mol or less, or 10 mol or less, or 8 mol or less, or 6 mol or less, or 4 mol or less, or 2 mol or less.
• the quantitative ratio of the fused ring dipeptide compound of formula (A1) to the protected amino acid or protected peptide of formula (R3) is not particularly limited, Protected amino acid or protected peptide, for example, 0.1 mol or more, or 0.2 mol or more, or 0.3 mol or more, or 0.4 mol or more, or 0.5 mol or more, and, for example, 20 mol or less, Alternatively, it can be used in a range of 15 mol or less, 10 mol or less, 8 mol or less, 6 mol or less, 4 mol or less, or 2 mol or less.
• the quantitative ratio of the fused ring dipeptide compound of formula (A1) to the amino acid ester or peptide ester of formula (R4) is not particularly limited, of amino acid ester or peptide ester, for example, 0.1 mol or more, or 0.2 mol or more, or 0.3 mol or more, or 0.4 mol or more, or 0.5 mol or more, and, for example, 20 mol or less, Alternatively, it can be used in a range of 15 mol or less, 10 mol or less, 8 mol or less, 6 mol or less, 4 mol or less, or 2 mol or less.
• the fused ring dipeptide compound of formula (A) as a substrate, the protected amino acid or protected peptide of formula (R3) , and the amino acid ester or peptide ester of formula (R4) must be used in an amount of 1 mole or more, respectively.
• the amount used is not particularly limited, but the base may be, for example, 0.2 mol or more, 0.4 mol or more, or 0.6 mol or more, per 1 mol of the fused ring dipeptide compound of formula (A1). mol or more, or 0.8 mol or more, or 1.0 mol or more, and also, for example, 40 mol or less, or 30 mol or less, or 20 mol or less, or 15 mol or less, or 10 mol or less, or 6 mol or less, or It can be used in a range of 4 moles or less.
• the amount used is not particularly limited, but the amount of the condensing agent is, for example, 0.2 mol or more, 0.4 mol or more, or 0. .6 moles or more, or 0.8 moles or more, or 1.0 moles or more, and also, for example, 40 moles or less, or 30 moles or less, or 20 moles or less, or 15 moles or less, or 10 moles or less, or 6 moles or less , or 4 moles or less.
• the amount used is not particularly limited, but for example, 0.2 mol or more of the racemization inhibitor per 1 mol of the fused ring dipeptide compound of formula (A1), or 0.4 mol or more, or 0.6 mol or more, or 0.8 mol or more, or 1.0 mol or more, and also, for example, 40 mol or less, or 30 mol or less, or 20 mol or less, or 15 mol or less, Alternatively, it can be used in a range of 10 mol or less, 6 mol or less, or 4 mol or less.
• reaction conditions in the second peptide production method of the present invention are not limited as long as the reaction proceeds, but examples are as follows for each reaction procedure for each of the first and second aspects. .
• the reaction conditions when adding and reacting the fused ring dipeptide compound of formula (A1) with the protected amino acid or protected peptide of formula (R3) in step (i) are such that the reaction proceeds.
• examples include the following.
• reaction temperature in step (i) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• the reaction pressure in step (i) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (i) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (i) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• reaction conditions for adding the fused ring dipeptide compound of formula (A2) to the reaction system and reacting are not limited as long as the reaction proceeds, and are, for example, as follows.
• reaction temperature in step (ii) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• reaction pressure in step (ii) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (ii) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (ii) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• reaction conditions when adding the amino acid ester or peptide ester of formula (R4) to the reaction system and reacting are not limited as long as the reaction proceeds, and are, for example, as follows.
• reaction temperature in step (iii) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• reaction pressure in step (iii) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (iii) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (iii) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• reaction conditions for adding and reacting the fused ring dipeptide compound of formula (A2) with the amino acid ester or peptide ester of formula (R4) in step (i) are as follows: Although there are no limitations as far as the process progresses, examples include the following.
• reaction temperature in step (i) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• the reaction pressure in step (i) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (i) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (i) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• reaction conditions for adding the fused ring dipeptide compound of formula (A1) to the reaction system and reacting are not limited as long as the reaction proceeds, and are, for example, as follows.
• reaction temperature in step (ii) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• reaction pressure in step (ii) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (ii) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (ii) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• reaction conditions for adding the protected amino acid or protected peptide of formula (R3) to the reaction system and reacting are not limited as long as the reaction proceeds, and are, for example, as follows.
• reaction temperature in step (iii) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• reaction pressure in step (iii) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (iii) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (iii) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• step (i), step (ii), and step (ii) may each be performed in a sequential method (batch method), or in a continuous method (flow method). (Act) may be implemented. Details of specific sequential (batch) and continuous (flow) method implementation procedures are known in the art. Further, step (i) and step (ii) and/or step (ii) and step (ii) may be performed continuously in one pod.
• ⁇ Polypeptide (target compound) The polypeptide that is the target compound finally produced in the second polypeptide production method of the present invention is a compound represented by the following formula (P2).
• R 111 , R 112 , R 113 , R 121 , and R 122 represent the same groups as defined in formula (A1) above, and R 211 , R 212 , R 213 , R 221 , and R 222 represents the same group as defined in the above formula (A2), and PG a , R 31 , R 32 , R 33 , A 31 , A 32 , p31, p32, and m are as defined in the above formula (R3).
• PG b , R 41 , R 42 , R 43 , A 41 , A 42 , p41, p42, and n represent the same group as defined in formula (R4) above.
• the compound of formula (R3) is a protected dipeptide and the compound of formula (R4) is an amino acid ester (that is, when m is 2 and n is 1)
• the compound of formula (R3) is
• the compound of formula (R4) is a protected amino acid and is a dipeptide ester (that is, m is 1 and n is 2)
• polypeptide compound of formula (P2) obtained by the above-mentioned production method may be further subjected to various post-treatments.
• post-treatments include isolation and purification of the obtained polypeptide compound of formula (P2), deprotection of the amino group-protecting group PG a and/or the carboxyl group-protecting group PG b , and the like. Such post-processing will be described in detail later.
• the fused ring tripeptide compound of the present invention is a silane-containing fused ring tripeptide compound having a structure represented by the following formula (B).
• R 11 , R 12 , R 13 , R 21 , R 22 , R a1 , and R a2 each independently represent a group having the same definition as the group with the same symbol in formula (A). .
• the details are as described above.
• R x1 and R x2 each independently have a hydrogen atom, a halogen atom, a hydroxyl group, a carboxyl group, a nitro group, a cyano group, or a thiol group, or one or more substituents.
• PG x represents a monovalent protecting group. Examples include, but are not limited to, monovalent aliphatic hydrocarbon groups, aromatic hydrocarbon groups, and heterocyclic groups that may have one or more substituents. Can be mentioned. In addition, when a substituent is present, the type thereof is as described above. Specific examples of the number of substituents are, for example, 5, 4, 3, 2, 1, or 0.
• the number of carbon atoms in the aliphatic hydrocarbon group is not particularly limited, but is within the upper limit. is, for example, 20 or less, 15 or less, 10 or less, 8 or less, or 6 or less.
• the lower limit varies depending on the type of aliphatic hydrocarbon group, but is 1 or more for alkyl groups, 2 or more for alkenyl or alkynyl groups, and 3 or more for cycloalkyl groups, such as 4 or more, or 5 or more. It is.
• Specific examples of the number of atoms include, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20. It is.
• the number of carbon atoms in the aromatic hydrocarbon group is not particularly limited, but is within the upper limit. is, for example, 20 or less, 15 or less, 10 or less, 8 or less, or 6 or less.
• the lower limit varies depending on the type of aromatic hydrocarbon group, but is usually 4 or more, for example 5 or more, or 6 or more.
• Specific examples of the number of atoms are, for example, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20.
• the fused ring tripeptide compound of the present invention can be produced by a method including the following steps (hereinafter appropriately referred to as "the fused ring tripeptide compound of the present invention").
• the fused ring tripeptide compound of the present invention (iv) A step of preparing a fused ring dipeptide compound of formula (A).
• a fused ring dipeptide compound of formula (A) (substrate compound):
• step (iv) a fused ring dipeptide compound of formula (A) is prepared.
• the method is not particularly limited, it is preferably produced by the method for producing the fused ring dipeptide compound of the present invention described above.
• step (v) the fused ring dipeptide compound of formula (A) prepared in step (iv) is reacted with an amino acid ester represented by formula (Rx) below.
• R x1 , R x2 , and PG x each independently represent a group having the same definition as the group with the same symbol in formula (B). The details are as described above.
• step (v) when reacting the fused ring dipeptide compound of formula (A) and the amino acid ester of formula (Rx), it is preferable to optionally coexist a fifth silane compound in the reaction system.
• the fifth silane compound When using the fifth silane compound, its type is not particularly limited, but examples include 1-(trimethylsilyl)imidazole (TMSIM), trimethylbromosilane (TMBS), trimethylchlorosilane (TMCS), tris(haloalkyl)silane , N-(trimethylsilyl)dimethylamine (TMSDMA), trimethylsilyl trifluoromethanesulfonate (TMS-OTf), dimethylsilylimidazole, dimethylsilyl(2-methyl)imidazole, dimethylethylsilylimidazole (DMESI), dimethylisopropylsilylimidazole (DMIPSI) ), 1-(tert-butyldimethylsilyl)imidazole (TBSIM), 1-(trimethylsilyl)triazole, 1-(tert-butyldimethylsilyl)triazole, N-methyl-Ntrimethylsilyltrifluoroacetamide (M
• step (v) other components may be present in the reaction system.
• other components include, but are not limited to, Lewis acid catalysts, bases, phosphoric acid, and the like. These details are as described above. Any one of these may be used alone, or two or more may be used in combination in any combination and ratio.
• the reaction may be carried out in a solvent.
• the solvent is not particularly limited, and examples thereof include aqueous solvents and organic solvents.
• Organic solvents include, but are not limited to, aromatic hydrocarbons such as toluene and xylene, pentane, petroleum ether, tetrahydrofuran (THF), 1-methyltetrahydrofuran (1-MeTHF), diisopropyl ether (i-Pr 2 O), ethers such as diethyl ether (Et 2 O) and cyclopentyl methyl ether (CPME), nitrogen-based organic solvents such as acetonitrile (MeCN), chlorine-based organic solvents such as dichloromethane (DCM), and ethyl acetate (AcOEt). and organic acids such as acetic acid. These solvents may be used alone or in combination of two or more.
• ⁇ Amount ratio of each ingredient is not particularly limited as long as the reaction is not inhibited.
• the amino acid ester of formula (Rx) may be added, for example, at least 0.1 mole, or at least 0.2 mole, or at least 0.3 mole, or at least 0.2 mole.
• the amount used is not particularly limited as long as it does not interfere with the reaction, but for example, the amount of the fifth silane compound is For example, 0.2 mole or more, or 0.4 mole or more, or 0.6 mole or more, or 0.8 mole or more, or 1.0 mole or more, and for example, 40 mole or less, or 30 mole or less. , or 20 mol or less, or 15 mol or less, or 10 mol or less, or 6 mol or less, or 4 mol or less.
• the total amount of the two or more types of fourth silane compounds may satisfy the above range.
• reaction conditions in step (v) are not limited as long as the reaction proceeds, but are exemplified as follows.
• the reaction temperature in step (v) is not limited as long as the reaction proceeds, but it is preferably carried out under heating conditions.
• the temperature can be, for example, 10°C or higher, 20°C or higher, 30°C or higher, 40°C or higher, or 50°C or higher.
• the upper limit is not particularly limited, but may be, for example, 120°C or lower, 110°C or lower, or 100°C or lower.
• reaction pressure in step (v) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (v) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (v) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• the fused ring tripeptide compound of the present invention obtained by the reaction in step (v) may be further subjected to various post-treatments.
• the produced fused ring tripeptide compound of the present invention can be isolated and purified according to conventional methods such as column chromatography and recrystallization.
• the produced fused ring dipeptide compound of the present invention may be used for producing a polypeptide, either directly or after isolation and purification, by subjecting it to the method for producing a polypeptide of the present invention described below.
• the fused ring tripeptide compound of the present invention can be used in various reactions, it is particularly suitable for use in the production of polypeptides.
• the method for producing a polypeptide using the fused ring tripeptide compound of the present invention includes two types of embodiments (these embodiments are hereinafter referred to as "the third method for producing the polypeptide of the present invention” and “the present method”). ). However, the method for producing a polypeptide using the fused ring tripeptide compound of the present invention is not limited to these two embodiments.
• the third method for producing a polypeptide of the present invention is a method for producing one molecule of a polypeptide compound of tetrapeptide or larger using one molecule of the fused ring tripeptide compound of the present invention, comprising the following step (vi).
• This is a method that includes (vi) By reacting the silane-containing condensed ring tripeptide compound represented by the above formula (B) with the protected amino acid or protected peptide compound represented by the following formula (Ra), a compound represented by the following formula (P3) is obtained.
• ⁇ Silane-containing fused ring tripeptide compound (substrate compound) In the third method for producing a polypeptide of the present invention, the silane-containing condensed ring tripeptide compound represented by the above formula (B) (the condensed ring tripeptide compound of the present invention) is used as a substrate compound. The details are as described above.
• the protected amino acid or protected peptide compound used as a substrate compound in the third method for producing a polypeptide of the present invention is a compound represented by the following formula (Ra).
• PG a represents a protecting group for an amino group. Details of PG a of formula (Ra) are as described above for PG a of formulas (R3) and (R4) in the first and second peptide production methods of the present invention.
• R a1 and R a2 each independently have a hydrogen atom, a halogen atom, a hydroxyl group, a carboxyl group, a nitro group, a cyano group, or a thiol group, or one or more substituents. represents an amino group, a monovalent aliphatic hydrocarbon group, a monovalent aromatic hydrocarbon group, or a monovalent heterocyclic group, which may be
• R a1 and R a2 in formula (Ra) are as described above for R 31 , R 32 , R 41 , and R 42 in formula (R3) and formula (R4).
• R a3 is a hydrogen atom, a carboxyl group, a hydroxyl group, a monovalent aliphatic hydrocarbon group that may have one or more substituents, an aromatic hydrocarbon group, or Represents a heterocyclic group.
• a monovalent aliphatic hydrocarbon group, aromatic hydrocarbon group, or heterocyclic group it may be bonded to the nitrogen atom via a linking group.
• R a1 and R a3 are bonded to each other to form a heterocycle which may have one or more substituents together with the carbon atom to which R a1 is bonded and the nitrogen atom to which R a3 is bonded. You can leave it there.
• the details of R a3 in formula (Ra) are as described above for R 33 and R 43 in formula (R3) and formula (R4).
• a a1 and A a2 each independently represent a divalent aliphatic hydrocarbon group having 1 to 3 carbon atoms which may have one or more substituents.
• the details of A a1 and A a2 in formula (Ra) are as described above for A 31 , A 32 , A 41 , and A 42 in formula (R3) and formula (R4).
• p a1 and p a2 each independently represent 0 or 1.
• m a is an integer of 1 or more and represents the number of structural units represented by the structure in [ ]. However, when m is 2 or more, the plurality of structural units represented by the structures in [ ] may be the same or different.
• the details of m a in formula (Ra) and the structural units in [ ] are as described above for m and n and the structural units in [ ] in formula (R3) and formula (R4).
• a base may also be allowed to coexist in the system from the viewpoint of increasing reaction efficiency.
• the type of base is not limited, and any known base known to improve reaction efficiency can be used.
• examples of such bases include those having 1 to 1 carbon atoms, such as tetrabutylammonium fluoride (TBAF), triethylamine (Et 3 N), diisopropylamine (i-Pr 2 NH), diisopropylethylamine (i-Pr 2 EtN), etc.
• examples include amines having 1 to 4 linear or branched alkyl groups of 10, and inorganic bases such as cesium fluoride. Any one of these may be used alone, or two or more may be used in combination in any combination and ratio.
• a condensing agent may be present in the system from the viewpoint of increasing the efficiency of the peptide formation reaction.
• a racemization inhibitor may be used in combination. The details of the condensing agent and the racemization inhibitor are as described above in the description of the first and second peptide production methods of the present invention.
• the substrate compound is the fused ring tripeptide compound of formula (B), the protected amino acid or protected peptide of formula (Ra), an optional base, a condensing agent, and
• other components may also be present. Examples include catalysts, silane compounds, phosphorus compounds, and the like. The details of these other components are also as explained in detail in the explanation of the first and second peptide production methods of the present invention.
• reaction may be carried out in a solvent.
• a solvent The details of such a solvent are also as described in detail in the explanation of the first and second peptide production methods of the present invention.
• the amounts of each component used are not limited, but are preferably as follows.
• the quantitative ratio of the fused ring tripeptide compound of formula (B) to the protected amino acid or protected peptide of formula (Ra) is not particularly limited, the ratio of the fused ring tripeptide compound of formula (B) to 1 mole of the fused ring tripeptide compound of formula ( Ra) protected amino acid or protected peptide, for example 0.1 mol or more, or 0.2 mol or more, or 0.3 mol or more, or 0.4 mol or more, or 0.5 mol or more, and for example 20 mol It can be used in a range of 15 moles or less, 10 moles or less, 8 moles or less, 6 moles or less, 4 moles or less, or 2 moles or less.
• the target production amount of the polypeptide compound of formula (P3) of the present invention to be manufactured is the fused ring tripeptide compound of formula (B) as a substrate and the protected amino acid or protected of formula (Ra). It is necessary to use 1 mole or more of each peptide.
• the amount used is not particularly limited, but the amount of the base is, for example, 0.2 mole or more, or 0.4 mole or more, or 0.2 mole or more, or 0.4 mole or more, or 0.2 mole or more, or 0.4 mole or more, per 1 mole of the fused ring tripeptide compound of formula (B). 6 moles or more, or 0.8 moles or more, or 1.0 moles or more, and also, for example, 40 moles or less, or 30 moles or less, or 20 moles or less, or 15 moles or less, or 10 moles or less, or 6 moles or less, Alternatively, it can be used in a range of 4 moles or less. In addition, when adding a base in a plurality of steps, it is preferable to add the base in an amount within the above range in each step.
• the amount used is not particularly limited, but the condensing agent may be, for example, 0.2 mol or more, or 0.4 mol or more, or 0.6 mole or more, or 0.8 mole or more, or 1.0 mole or more, and also, for example, 40 mole or less, or 30 mole or less, or 20 mole or less, or 15 mole or less, or 10 mole or less, or 6 mole It can be used in a range of 4 moles or less.
• the amount used is not particularly limited, but the amount of the racemization inhibitor is, for example, 0.2 mol or more per 1 mol of the fused ring tripeptide compound of formula (B). , or 0.4 mol or more, or 0.6 mol or more, or 0.8 mol or more, or 1.0 mol or more, and also, for example, 40 mol or less, or 30 mol or less, or 20 mol or less, or 15 mol or less , or 10 mol or less, 6 mol or less, or 4 mol or less.
• step (vi) the fused ring tripeptide compound of formula (B) is reacted with the protected amino acid or protected peptide compound of formula (Ra).
• the reaction conditions are not limited as long as the reaction proceeds, but are exemplified as follows.
• reaction temperature in step (vi) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• reaction pressure in step (vi) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (vi) is also not limited as long as the reaction proceeds, but it can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (vi) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• the polypeptide compound that is the target compound finally produced in the third polypeptide production method of the present invention is a compound represented by the following formula (P3). (P3)
• PG a , R a1 , R a2 , R a3 , A a1 , A a2 , p a1 , p a2 , and m a are groups with the same definition as the groups with the same symbols in the formula (Ra) above. and R 11 , R 12 , R 13 , R 21 , R 22 , R x1 , R x2 , and PG x represent groups having the same definition as the groups with the same symbols in the formula (B).
• polypeptide compound of formula (P3) obtained by the above-mentioned production method may be further subjected to various post-treatments.
• post-treatments include isolation and purification of the obtained polypeptide compound of formula (P3), deprotection of the amino group-protecting group PG a and/or the carboxyl group-protecting group PG x , and the like. Such post-processing will be described in detail later.
• the fourth method for producing a polypeptide of the present invention is a method for producing one molecule of a hexapeptide compound using two molecules of the fused ring tripeptide compound of the present invention. Further, optionally, by further reacting the obtained hexapeptide compound with a protected amino acid or a protected peptide compound, it is also possible to produce a polypeptide compound of heptapeptide or higher.
• the fourth method for producing a polypeptide of the present invention is a method that includes at least the following steps (vii) and (viii). Furthermore, the following step (ix) may be optionally included.
• (vii) A step of mixing a silane-containing condensed ring tripeptide compound represented by the following formula (B1) with a base.
• (viii) A step of obtaining a hexapeptide compound represented by the following formula (P4) by reacting the mixture of step (vii) with a silane-containing condensed ring tripeptide compound represented by the following formula (B2).
• the silane-containing condensed ring tripeptide compound used as a substrate compound in the method for producing the fourth polypeptide of the present invention is the silane-containing condensed ring tripeptide compound represented by the above formula (B) (the condensed ring tripeptide compound of the present invention).
• B the condensed ring tripeptide compound of the present invention.
• the compound is similar to the above-mentioned third polypeptide production method of the present invention, in that two molecules of a silane-containing condensed ring tripeptide compound are used when synthesizing one molecule of polypeptide. is different.
• the compound on the nucleophile side is represented by the following formula (B1)
• the compound on the electrophile side is represented by the following formula (B2).
• R 111 , R 112 , R 113 , R 211 , R 212 , R a11 , R a12 , R x11 , and R x12 are R 11 , R 12 , and R 13 in formula (B), respectively.
• R 21 , R 22 , R a1 , R a2 , R x1 , and R x2 are R 11 , R 12 , and R 13 in formula (B), respectively.
• PG x1 represents a carboxyl group protecting group, like PG x in formula (B).
• the carboxyl protecting group PG x of formula (B1) which is a compound on the nucleophile side, is not limited to, but includes, but is not limited to, a methyl group, an ethyl group, a propyl group, an isopropyl group, an n-butyl group, an isobutyl group, Alkyl groups such as tert-butyl group, sec-butyl group, pentyl group, isopentyl group, neopentyl group, hexyl group, heptyl group, octyl group, decyl group, nonyl group; Included are substituted haloaryl groups and haloarylalkyl groups.
• R 121 , R 122 , R 123 , R 221 , R 222 , R a21 , R a22 , R x21 , and R x22 are R 11 , R 12 , and R 13 in formula (B), respectively.
• R 21 , R 22 , R a1 , R a2 , R x1 , and R x2 are R 11 , R 12 , and R 13 in formula (B), respectively.
• R x23 represents -O-PG x , -NH-PG x or -S-PG x .
• PG x represents a monovalent protecting group having the same definition as PG x in the above formula (B).
• PG x is as described above.
• the protecting group PG x of formula (B2) which is an electrophilic species, includes alkynyl groups such as propargyl group; phenyl group, benzyl group, tolyl group, cumyl group, 1,1-diphenylethyl group, triphenylmethyl group.
• aryl groups and arylalkyl groups such as fluorenyl groups, naphthyl groups, and anthracenyl groups; haloaryl groups and haloarylalkyl groups such as pentafluorophenyl groups in which the aryl groups and arylalkyl groups are substituted with one or more halogen groups; Group; trimethylsilyl (TMS) group, triethylsilyl (TES) group, triisopropylsilyl (TIPS) group, tri-tert-butylsilyl (TBS) group, tert-butyldiphenylsilyl (TBDPS) group, tris(trialkylsilyl)silyl group Silicon-based protecting groups such as; and the like are preferred.
• TMS trimethylsilyl
• TES triethylsilyl
• TIPS triisopropylsilyl
• TBS tri-tert-butylsilyl
• TDPS
• a base is used in step (vii).
• the type of base is not limited, and any known base known to improve reaction efficiency can be used.
• examples of such bases include those having 1 to 1 carbon atoms, such as tetrabutylammonium fluoride (TBAF), triethylamine (Et 3 N), diisopropylamine (i-Pr 2 NH), diisopropylethylamine (i-Pr 2 EtN), etc.
• examples include amines having 1 to 4 linear or branched alkyl groups of 10, and inorganic bases such as cesium fluoride. Any one of these may be used alone, or two or more may be used in combination in any combination and ratio.
• a condensing agent may be present in the system from the viewpoint of increasing the efficiency of the peptide formation reaction.
• a racemization inhibitor may be used in combination. The details of the condensing agent and the racemization inhibitor are as detailed above in the description of the first to third peptide production methods of the present invention.
• the fused ring tripeptide compounds of the aforementioned formulas (B1) and (B2) which are substrate compounds, and a base, as well as an optional condensing agent and a racemization inhibitor, are used.
• other components may also be present. Examples include catalysts, silane compounds, phosphorus compounds, and the like. The details of these other components are also as detailed above in the description of the first to third peptide production methods of the present invention.
• reaction may be carried out in a solvent.
• a solvent The details of such a solvent are also as detailed above in the explanation of the first to third peptide production methods of the present invention.
• step (vii) a fused ring tripeptide compound of formula (B1), which is a substrate compound on the nucleophile side, is mixed with a base.
• step (viii) the mixture of step (vii) is mixed with the fused ring tripeptide compound of formula (B2), which is the substrate compound on the electrophilic species side.
• the fused ring tripeptide compound of formula (B1) opens the ring and functions as a nucleophilic species
• the fused ring tripeptide compound of formula (B2) opens the ring and functions as an electrophilic species, forming an amide bond.
• a hexapeptide compound of formula (P4) will be obtained.
• timing of adding optionally used other components such as a condensing agent to the reaction system is not particularly limited, and they may be added at any timing. However, when using a condensing agent and/or a base, it is preferable to add it to the system at the beginning of step (vii) and/or step (viii). Furthermore, when a racemization inhibitor is used in addition to the condensing agent, it is preferably added to the system together with the condensing agent. Furthermore, when the reaction is carried out using a solvent, the components may be mixed in the solvent and brought into contact with each other.
• the amounts of each component used are not limited, but are preferably as follows.
• the ratio of the fused ring tripeptide compound of formula (B1) to the fused ring tripeptide compound of formula (B2) is not particularly limited, but the ratio of the fused ring tripeptide compound of formula (B1) to 1 mole of the fused ring tripeptide compound of formula (B1) is B2) fused ring tripeptide compound, for example 0.05 mol or more, or 0.1 mol or more, or 0.2 mol or more, or 0.3 mol or more, or 0.4 mol or more, or 0.5 mol In addition, it can be used in a range of, for example, 20 mol or less, 15 mol or less, 10 mol or less, 8 mol or less, 6 mol or less, 4 mol or less, or 2 mol or less.
• the amount of the base to be used is not particularly limited, but the base may be, for example, 0.2 mol or more, or 0.4 mol or more, or 0.6 mol or more, or 0.8 mol or more, or 1.0 mol or more, and also, for example, 40 mol or less, or 30 mol or less, or 20 mol or less, or 15 mol or less, or 10 mol or less, or 6 mol or less, or 4 mol or less Can be used within a range.
• the amount used is not particularly limited, but the condensing agent may be, for example, 0.2 mol or more, or 0.4 mol or more, or 0.6 mole or more, or 0.8 mole or more, or 1.0 mole or more, and also, for example, 40 mole or less, or 30 mole or less, or 20 mole or less, or 15 mole or less, or 10 mole or less, or 6 mole It can be used in a range of 4 moles or less.
• the amount used is not particularly limited, but the amount of the racemization inhibitor is, for example, 0.2 mol or more per 1 mol of the fused ring tripeptide compound of formula (B1). , or 0.4 mol or more, or 0.6 mol or more, or 0.8 mol or more, or 1.0 mol or more, and also, for example, 40 mol or less, or 30 mol or less, or 20 mol or less, or 15 mol or less , or 10 mol or less, 6 mol or less, or 4 mol or less.
• reaction conditions in the fourth peptide production method of the present invention are not limited as long as the reaction proceeds, but examples for each reaction procedure are as follows.
• reaction conditions when mixing the fused ring tripeptide compound of formula (B1) and a base are not limited as long as the reaction proceeds, but are, for example, as follows.
• reaction temperature in step (vii) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• reaction pressure in step (vii) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (vii) is also not limited as long as the reaction proceeds, but the reaction can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (vii) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• step (viii) the reaction conditions for adding the fused ring tripeptide compound of formula (B2) to the mixture of step (vii) and reacting are not limited as long as the reaction proceeds, but for example, the following. That's right.
• reaction temperature in step (viii) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• reaction pressure in step (viii) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (viii) is also not limited as long as the reaction proceeds, but it can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (viii) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• step (vii) and step (viii) may each be carried out in a sequential method (batch method) or in a continuous method (flow method). Details of specific sequential (batch) and continuous (flow) method implementation procedures are known in the art. Further, step (vii) and step (viii) may each be performed continuously in one pod.
• ⁇ Hexapeptide (target compound) In the fourth method for producing a polypeptide of the present invention, the hexapeptide compound that is the target compound produced after step (viii) is a compound represented by the following formula (P4).
• R 111 , R 112 , R 113 , R 211 , R 212 , R x11 , R x12 , and PG x1 represent groups having the same definition as the groups with the same symbols in formula (B1)
• R 121 , R 122 , R 123 , R 221 , R 222 , R x21 , and R x22 represent groups having the same definition as the groups with the same symbols in the formula (B2).
• the hexapeptide compound of formula (P4) obtained by the above-mentioned production method may be further subjected to various post-treatments.
• Such post-treatments include isolation and purification of the obtained hexapeptide compound of formula (P4), deprotection of the carboxyl group protecting group PG x1 , and the like. Such post-processing will be described in detail later.
• step (ix) the hexapeptide compound of formula (P4) obtained in step (viii) is Mix the protected amino acid or protected peptide compound.
• the amount of the protected amino acid or protected peptide compound of formula (Ra) to be used is not particularly limited, but the base may be, for example, 0.2 mol or more, or 0.4 mol, per 1 mol of the fused ring tripeptide compound of formula (B1). or more, or 0.6 mol or more, or 0.8 mol or more, or 1.0 mol or more, and also, for example, 40 mol or less, or 30 mol or less, or 20 mol or less, or 15 mol or less, or 10 mol or less, Alternatively, it can be used in a range of 6 mol or less, or 4 mol or less.
• reaction conditions in step (ix) are limited as long as the reaction proceeds.
• examples are as follows.
• reaction temperature in step (ix) is not limited as long as the reaction proceeds, but is, for example, 0°C or higher, or 10°C or higher, or 20°C or higher, or, for example, 100°C or lower, or 80°C or lower, or 60°C or lower. It can be done.
• reaction pressure in step (ix) is also not limited as long as the reaction proceeds, and may be carried out under reduced pressure, normal pressure, or increased pressure, but usually it can be carried out at normal pressure.
• reaction atmosphere in step (ix) is also not limited as long as the reaction proceeds, but it can be carried out in an atmosphere of an inert gas such as argon or nitrogen.
• reaction time of step (ix) is also not limited as long as the reaction proceeds, but from the viewpoint of allowing the reaction to proceed sufficiently and efficiently, for example, 10 minutes or more, or 20 minutes or more, or 30 minutes or more, for example. It can be within 80 hours, or within 60 hours, or within 50 hours.
• steps (vii) and (viii) and step (ix) may be performed in a sequential method (batch method). It may also be carried out using a continuous method (flow method). Details of specific sequential (batch) and continuous (flow) method implementation procedures are known in the art. Further, steps (vii) and (viii) and step (ix) may be performed consecutively in one pod.
• the polypeptide compound of heptapeptide or higher which is the target compound produced after step (ix), is a compound represented by the following formula (P5). be.
• PG a , R a1 , R a2 , R a3 , A a1 , A a2 , p a1 , p a2 , and m a represent groups having the same definition as the groups with the same symbols in the formula (Ra), and R 111 , R 112 , R 113 , R 211 , R 212 , R x11 , R x12 , PG x1 , R 121 , R 122 , R 123 , R 221 , R 222 , R x21 , and R x22 in the formula (P4) Represents a group with the same sign and a group with the same definition. Further, the encircled symbol A at the right end of the upper structure and the left end of the lower structure in formula (P5) means that the upper structure and the lower structure are continuous at this position.
• polypeptide compound of formula (P5) obtained by the above-mentioned production method may be further subjected to various post-treatments.
• post-treatments include isolation and purification of the obtained polypeptide compound of formula (P5), deprotection of the amino group-protecting group PG a and/or the carboxyl group-protecting group PG b , and the like. Such post-processing will be described in detail later.
• polypeptide compounds of formulas (P1) to (P5) obtained by the above production method may be further subjected to various post-treatments.
• polypeptide compounds of formulas (P1) to (P5) obtained by the above-mentioned production method can be isolated and purified according to conventional methods such as column chromatography and recrystallization.
• the amino group protected by the protecting group PG a can also be deprotected.
• the method for deprotecting the protected amino group is not particularly limited, and various methods can be used depending on the type of protecting group PG a . Examples include deprotection by hydrogenation, deprotection by weak acids, deprotection by fluorine ions, deprotection by one-electron oxidizing agents, deprotection by hydrazine, deprotection by oxygen, and the like.
• deprotection by hydrogenation In the case of deprotection by hydrogenation, (a) deprotection by reduction in the presence of hydrogen gas using a metal catalyst such as palladium, palladium-carbon, palladium hydroxide, palladium hydroxide-carbon, etc. as a reduction catalyst; (b) In the presence of a metal catalyst such as palladium, palladium-carbon, palladium hydroxide, palladium hydroxide-carbon, etc., sodium borohydride, lithium aluminum hydride, lithium borohydride, diborane, etc. Examples include a method of reducing and deprotecting using a hydrogenation reducing agent.
• a metal catalyst such as palladium, palladium-carbon, palladium hydroxide, palladium hydroxide-carbon, etc.
• sodium borohydride lithium aluminum hydride, lithium borohydride, diborane, etc. Examples include a method of reducing and deprotecting using a hydrogenation reducing agent.
• the carboxyl group protected by the protecting group PG b can also be deprotected.
• the method for deprotecting the protected carboxyl group is not particularly limited, and various methods can be used depending on the type of the protecting group PG b . Examples include deprotection by hydrogenation, deprotection by base, deprotection by weak acid, and the like. In the case of deprotection using a base, examples include a method of deprotecting using a strong base such as lithium hydroxide, sodium hydroxide, potassium hydroxide, etc. as the base.
• polypeptide compounds of formulas (P1) to (P5) obtained by the above-mentioned production method can be used to obtain the protected peptide of formula (R3) or formula (Ra) and / Or it may be used as a peptide ester of formula (R4) and subjected again to the first to fourth peptide production methods of the present invention.
• the polypeptide compounds of formulas (P1) to (P5) obtained by the above-mentioned production method can be subjected to other conventionally known amidation methods or peptide production methods. Good too.
• polypeptides can be synthesized by linking other amino acids or peptides to the polypeptide compounds of formulas (P1) to (P5) through amide bonds and elongating the amino acid residues. By successively repeating these steps, it is theoretically possible to synthesize polypeptides with any number of amino acid residues and any amino acid sequence.
• Patent Document 4 mentioned above
• Patent Document 5 International Publication No. 2021/085635
• Patent Document 6 International Publication No. 2021/085636
• Patent Document 7 International Publication No. 2021/149814
• Patent Document 8 International Publication No. 2022/190486
• a silane-containing condensed ring dipeptide compound of formula (A) shown in the table below (1 equivalent; 0.25 mmol) synthesized by the method described in Example 1 and an amino acid ester of formula (Rx) (3 equivalents) were placed in a 20 mL test tube.
• TMS-IM trimethylsilylimidazole
• a silane-containing condensed ring dipeptide compound shown in the above reaction formula (a compound in which PG x1 is Me in formula (B1): 1 equivalent) synthesized by the method described in Example 2, and 1 M tetrabutylammonium fluoride were placed in a 20 mL test tube.
• a solution of TBAF (base; 1 equivalent) in tetrahydrofuran (THF) and dichloromethane (DCM; 1.5 mL) were added, and the mixture was stirred at 50° C. for 24 hours.
• a silane-containing condensed ring dipeptide compound shown in the above reaction formula (a compound in which PG x1 is Me in formula (B1): 1 equivalent) synthesized by the method described in Example 2, and 1 M tetrabutylammonium fluoride were placed in a 20 mL test tube.
• a solution of TBAF (base; 1 equivalent) in tetrahydrofuran (THF) and dichloromethane (DCM; 1.5 mL) were added, and the mixture was stirred at 50° C. for 24 hours.

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