Method for producing lipid peptides

Abstract

[Problem] To provide a method by which a lipid peptide compound and a salt thereof can be produced with a higher yield and a higher purity than conventional production methods. [Solution] A method for producing a lipid peptide compound represented by formula (3) or a pharmaceutically usable salt thereof, said method comprising: a reaction step for reacting an ester compound represented by formula (1) with an α-amino acid compound represented by formula (2) and a base in a solvent containing a nonpolar organic solvent; an extraction step for neutralizing the solution, which is obtained in the reaction step and in which a salt of the lipid peptide represented by formula (3) is dissolved, by adding an organic acid thereto, then adding water and an alcohol for liquid separation and removing the nonpolar organic solvent; and a separation step for taking out the lipid peptide compound represented by formula (3) from the solution, after the extraction step, to outside the system.

Description

[Technical Field]

[0001] The present invention relates to a method for producing a lipid peptide. [Background technology]

[0002] In recent years, the use of novel lipid peptides, in which glycine or histidine is bound to palmitic acid or the like, as hydrogelators has been proposed, and the method for supplying these compounds has become important (Patent Document 1). On the other hand, solid-phase peptide synthesis is generally proposed as a method for producing lipid peptides, but it is only suitable for small-scale synthesis and large-scale production is difficult.

[0003] On the other hand, the present inventors have previously reported that when the amino group of an amino acid is amidated with an ester compound, a lipid peptide compound can be directly obtained without using a protecting group by reacting the compound in a solvent containing a nonpolar organic solvent in the presence of a base (Patent Document 2). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] International Publication No. 2010 / 013555 [Patent Document 2] International Publication No. 2011 / 027897 Summary of the Invention [Problem to be solved by the invention]

[0005] The method described in Patent Document 2 is extremely advantageous compared to conventional production methods in that it does not require more complicated operations and allows for the production of practical lipid peptide compounds that can be mass-produced inexpensively. However, there is a need to develop a method for producing lipid peptide compounds in high yield and high purity, which is suitable for further industrial production. The present invention has been made in consideration of the above circumstances, and its object is to provide a method for producing lipid peptide compounds with higher yield and higher purity than conventional production methods. [Means for solving the problem]

[0006] In order to achieve the above object, the present inventors have conducted extensive research focusing on a method for removing the nonpolar organic solvent after the reaction is completed and before neutralization, and on the raw materials. As a result, they have found that by adopting a method for removing the nonpolar organic solvent in which an organic acid is added to the solution after the reaction is completed to neutralize it, then adding water and alcohol to separate the liquids, and then removing the nonpolar organic solvent, and by using a stable ethyl ester as the raw material, the amount of the raw material can be reduced, the reaction can be carried out in a homogeneous system, and lipid peptide compounds can be produced in higher yields and with higher purity, thereby completing the present invention.

[0007] That is, in a first aspect, the present invention provides a compound represented by formula (1): [ka] (In the formula, R 1 represents an aliphatic group having a linear or branched chain structure and having 9 to 23 carbon atoms; R 2 represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms, which may have a branched chain having 1 or 2 carbon atoms; R 3 represents an alkyl group having 1 to 6 carbon atoms, a haloalkyl group having 1 to 6 carbon atoms, a hydroxyalkyl group having 1 to 6 carbon atoms, or an aryl group which may be substituted with an alkyl group having 1 to 6 carbon atoms. [ka] (In the formula, R 4 Ha-(CH2) na reaction step of reacting an α-amino acid compound represented by the formula (I) with a base in a solvent containing a nonpolar organic solvent, the reaction mixture being represented by the formula (I): The compound of formula (3) obtained in this reaction step [ka] (In the formula, R 1 , R 2 and R 4 represents the above-defined meaning. An extraction step in which an organic acid is added to a solution in which a salt of a lipid peptide represented by the formula (I) is dissolved, the solution is neutralized, water and alcohol are added, the solution is separated, and the non-polar organic solvent is removed; The present invention relates to a method for producing a lipid peptide compound represented by formula (3) or a pharmaceutically usable salt thereof, which comprises a separation step of removing the lipid peptide compound represented by formula (3) from the solution after the extraction step. As a second aspect, the present invention relates to the production method according to the first aspect, wherein the solvent contains a non-polar organic solvent and an alcohol. As a third aspect, the present invention relates to the production method according to the first aspect, wherein, in the formula, n represents a number of 1 to 4 and X represents an amino group, a guanidino group, or a -CONH group, or n represents 1 and X represents a pyrrole group, an imidazole group, a pyrazole group, or an imidazole group. As a fourth aspect, in the formula, R 1 represents a linear aliphatic group having 11 to 21 carbon atoms and which may have 0 to 2 unsaturated bonds. As a fifth aspect, in the formula, R 2 represents a hydrogen atom or an alkyl group having 1 to 3 carbon atoms, which may have a branched chain having 1 carbon atom. As a sixth aspect, in the formula, R 2 represents a hydrogen atom, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group, or a tert-butyl group; R 4represents an aminomethyl group, an aminoethyl group, a 3-aminopropyl group, a 4-aminobutyl group, a carbamoylmethyl group, a 2-carbamoylethyl group, a 3-carbamoylbutyl group, a 2-guanidinoethyl group, a 3-guanidinopropyl group, a pyrrolemethyl group, an imidazolemethyl group, a pyrazolemethyl group or a 3-indolemethyl group. As a seventh aspect, in the formula, R 2 represents a hydrogen atom, a methyl group, an isopropyl group, an isobutyl group, or a sec-butyl group, and R 4 represents a 4-aminobutyl group, a carbamoylmethyl group, a 2-carbamoylethyl group, a 3-guanidinopropyl group, an imidazolemethyl group, or a 3-indolemethyl group. As an eighth aspect, the present invention relates to the production method according to the first aspect, in which the organic acid is acetic acid. As a ninth aspect, the present invention relates to the production method according to any one of the first to eighth aspects, in which the base is at least one selected from an alkali metal, an alkali metal inorganic acid salt, an alkali metal hydroxide, an alkali metal alkoxide, an alicyclic amine, an alcohol solution thereof, or an alcohol dispersion thereof. According to a tenth aspect, the present invention provides the production method according to the ninth aspect, wherein the base is at least one selected from metallic sodium, metallic potassium, sodium carbonate, potassium carbonate, potassium phosphate, sodium phosphate, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, potassium methoxide, potassium ethoxide, potassium t-butoxide, 1,8-diazabicyclo[5.4.0]-7-undecene, 1,5-diazabicyclo[4.3.0]-5-nonene, an alcohol solution thereof, or an alcohol dispersion thereof. As an eleventh aspect, the present invention relates to the production method according to the tenth aspect, in which the base is sodium methoxide, a methanol solution thereof, or a methanol dispersion thereof. As a twelfth aspect, the present invention relates to the production method according to any one of the first to eleventh aspects, in which the nonpolar organic solvent is at least one selected from the group consisting of aromatic compounds, saturated aliphatic compounds, and unsaturated aliphatic compounds. According to a thirteenth aspect, the present invention relates to the production method according to the twelfth aspect, in which the non-polar organic solvent is at least one selected from the group consisting of toluene, xylene, ortho-dichlorobenzene, pentane, hexane, heptane, octane, cyclopentane, cyclohexane, methylcyclohexane, cycloheptane, and 1-hexene. According to a fourteenth aspect, in the production method according to the second aspect, the solvent contains toluene and methanol or ethanol. According to a fifteenth aspect, the production method according to any one of the first to fourteenth aspects, wherein the reaction of the ester compound represented by formula (1) with the α-amino acid compound represented by formula (2) is carried out at a reaction temperature of 65°C to 75°C. As a 16th viewpoint, Equation (1) [ka] (In the formula, R 1 represents an aliphatic group having a linear or branched chain structure and having 9 to 23 carbon atoms; R 2 represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms, which may have a branched chain having 1 or 2 carbon atoms; R 3 represents an alkyl group having 1 to 6 carbon atoms, a haloalkyl group having 1 to 6 carbon atoms, a hydroxyalkyl group having 1 to 6 carbon atoms, or an aryl group which may be substituted with an alkyl group having 1 to 6 carbon atoms. [ka] (In the formula, R 4 Ha-(CH2) na reaction step of reacting an α-amino acid compound represented by the formula (I) with a base in a solvent containing a nonpolar organic solvent, the reaction mixture being represented by the formula (I): The compound of formula (3) obtained in this reaction step [ka] (In the formula, R 1 , R 2 and R 4 represents the above-defined meaning. An extraction step of adding an organic acid to a solution in which a salt of a lipid peptide represented by (I) is dissolved to neutralize the solution, then adding water and alcohol to separate the solution, and removing the non-polar organic solvent; a pH adjustment step of adjusting the pH of the solution after the extraction step using hydrogen halide; and The present invention relates to a method for producing a lipid peptide compound represented by formula (3), which comprises a separation step of removing the lipid peptide compound represented by formula (3) from the solution after the pH adjustment step. The seventeenth point is: Formula (4) [ka] (wherein X is a halogen atom, an alkoxy group having 1 to 6 carbon atoms, or —OC(O)R 1 represents a group, and R 1 represents an aliphatic group having a linear or branched chain structure and having 9 to 23 carbon atoms, and a compound represented by formula (5) [ka] (In the formula, R 2 represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms, which may have a branched chain having 1 or 2 carbon atoms; R 3 represents an alkyl group having 1 to 6 carbon atoms, a haloalkyl group having 1 to 6 carbon atoms, a hydroxyalkyl group having 1 to 6 carbon atoms, or an aryl group which may be substituted with an alkyl group having 1 to 6 carbon atoms. [ka] (In the formula, R 1 , R 2 and R 3 represents the same as defined above. The ester compound represented by formula (1) obtained in the production step and the ester compound represented by formula (2) [ka] (In the formula, R 4 is a hydrogen atom, an alkyl group having 1 to 7 carbon atoms which may have a branched chain and which has 1 to 3 carbon atoms, a phenylmethyl group, a phenylethyl group, -(CH2) n a reaction step of reacting an α-amino acid compound represented by the formula (I) with a base in a solvent containing a nonpolar organic solvent, the reaction mixture being represented by the formula (I): The compound of formula (3) obtained in this reaction step [ka] (In the formula, R 1 , R 2 and R 4 represents the above-defined meaning. An extraction step in which an organic acid is added to a solution in which a salt of a lipid peptide represented by the formula (I) is dissolved, the solution is neutralized, water and alcohol are added, the solution is separated, and the non-polar organic solvent is removed; The present invention relates to a method for producing a lipid peptide compound represented by formula (3) or a pharmaceutically usable salt thereof, which comprises a separation step of removing the lipid peptide compound represented by formula (3) from the solution after the extraction step. According to an eighteenth aspect, the present invention relates to the production method according to the seventeenth aspect, in which the reaction of the compound represented by formula (4) with the compound represented by formula (5) is carried out in a homogeneous phase at a reaction temperature of 35°C to 45°C. [Effects of the Invention]

[0008] The production method of the present invention has the advantage that the desired lipid peptide compound and its salt can be obtained with high purity and in high yield, and is suitable for industrial production. DETAILED DESCRIPTION OF THE INVENTION

[0009] In this specification, "n" means normal, "i" means iso, "s" or "sec" means secondary, "t" or "tert" means tertiary, "c" means cyclo, "o" means ortho, "m" means meta, "p" means para, "Me" means methyl group, "Bu" means butyl group, and "tBu" means tertiary butyl group.

[0010] In the above formula (1), R 1 represents an aliphatic group having 9 to 23 carbon atoms, and preferably R 1 is preferably a linear aliphatic group having 11 to 21 carbon atoms or a linear aliphatic group having 11 to 21 carbon atoms and one or two unsaturated bonds.

[0011] Here, particularly preferred R 1 Specific examples of the aliphatic group represented by the formula (I) include a nonyl group, a decyl group, an undecyl group, a dodecyl group (lauryl group), a tridecyl group, a tetradecyl group (myristyl group), a pentadecyl group, a hexadecyl group (palmityl group), a heptadecyl group (margaryl group), an octadecyl group (stearyl group), a nonadecyl group, an icosyl group, and a heneicosyl group.

[0012] In the above formula (1), R 2 represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms which may have a branched chain and which has 1 or 2 carbon atoms.

[0013] Above R 2In the formula (I), the alkyl group having 1 to 4 carbon atoms, which may have a branched chain having 1 or 2 carbon atoms, means an alkyl group having 1 to 4 carbon atoms in the main chain and which may have a branched chain having 1 or 2 carbon atoms, and specific examples thereof include a methyl group, an ethyl group, an n-propyl group, an i-propyl group, an n-butyl group, an i-butyl group, a sec-butyl group, and a tert-butyl group.

[0014] R 2 is preferably a hydrogen atom or an alkyl group of 1 to 3 carbon atoms which may have a branched chain of 1 carbon atom, and more preferably a hydrogen atom. The alkyl group of 1 to 3 carbon atoms which may have a branched chain of 1 carbon atom means an alkyl group having 1 to 3 carbon atoms in the main chain and which may have a branched chain of 1 carbon atom, and specific examples thereof include a methyl group, an ethyl group, an n-propyl group, an i-propyl group, an i-butyl group, and a sec-butyl group, and preferably a methyl group, an i-propyl group, an i-butyl group, or a sec-butyl group.

[0015] In the above formula (1), R 3 represents an alkyl group having 1 to 6 carbon atoms, a haloalkyl group having 1 to 6 carbon atoms, a hydroxyalkyl group having 1 to 6 carbon atoms, or an aryl group optionally substituted with an alkyl group having 1 to 6 carbon atoms, preferably an alkyl group having 1 to 6 carbon atoms, and more preferably an ethyl group.

[0016] In the above formula (2), R 4 Ha-(CH2) n represents the -X group. Above -(CH2) n In the -X group, n represents a number from 1 to 4, and X represents an amino group, a guanidino group, a -CONH2 group, or a 5- or 6-membered ring or a fused heterocycle composed of a 5- and 6-membered ring, which may have 1 to 3 nitrogen atoms.

[0017] Above -(CH2) nIn the -X group, X preferably represents an amino group, a guanidino group, a -CONH2 group, a pyrrole group, an imidazole group, a pyrazole group, or an indole group, and more preferably an imidazole group. n In the -X group, n is preferably 1 or 2, more preferably 1. Therefore, the above -(CH2) n The -X group preferably represents an aminomethyl group, a 2-aminoethyl group, a 3-aminopropyl group, a 4-aminobutyl group, a carbamoylmethyl group, a 2-carbamoylethyl group, a 3-carbamoylbutyl group, a 2-guanidinoethyl group, a 3-guanidinopropyl group, a pyrrolemethyl group, an imidazolemethyl group, a pyrazolemethyl group, or a 3-indolemethyl group, more preferably a 4-aminobutyl group, a carbamoylmethyl group, a 2-carbamoylethyl group, a 3-guanidinopropyl group, an imidazolemethyl group, or a 3-indolemethyl group, and even more preferably an imidazolemethyl group.

[0018] Therefore, among the lipid peptide compounds represented by the above formula (3), particularly preferred specific examples include compounds formed from the following lipid moiety and dipeptide moiety: Histidine (His), glycine (Gly), valine (Val), isoleucine (Ile), alanine (Ala), arginine (Arg), asparagine (Asn), glutamine (Gln), leucine (Leu), lysine (Lys), and tryptophan (Trp) are used as abbreviations for amino acids. : N-Lauroyl-Gly-His, N-Lauroyl-Gly-Trp, N-Lauroyl-Gly-Gln, N-Lauroyl-Gly-Asn, N-Lauroyl-Gly-Arg, N-Lauroyl-Gly-Lys, N-Lauroyl-Ala-His, N-Lauroyl-Ala-Trp, N-Lauroyl-Ala-Gln, N-Lauroyl-Ala-Asn, N-Lauroyl-Ala-Arg, N-Lauroyl-Ala-Lys, N-Lauroyl N-Lauroyl-Val-His, N-Lauroyl-Val-Trp, N-Lauroyl-Val-Gln, N-Lauroyl-Val-Asn, N-Lauroyl-Val-Arg, N-Lauroyl-Val-Lys, N-Lauroyl-Leu-His, N-Lauroyl-Leu-Trp, N-Lauroyl-Leu-Gln, N-Lauroyl-Leu-Asn, N-Lauroyl-Leu-Arg, N-Lauroyl-Leu-Lys, N-Lauroyl- Ile-His, N-Lauroyl-Ile-Trp, N-Lauroyl-Ile-Gln, N-Lauroyl-Ile-Asn, N-Lauroyl-Ile-Arg, N-Lauroyl-Ile-Lys, N-Myristoyl-Gly-His, N-Myristoyl-Gly-Trp, N-Myristoyl-Gly-Gln, N-Myristoyl-Gly-Asn, N-Myristoyl-Gly-Arg, N-Myristoyl-Gly-Lys, N-Myristoyl N-myristoyl-Ala-His, N-myristoyl-Ala-Trp, N-myristoyl-Ala-Gln, N-myristoyl-Ala-Asn, N-myristoyl-Ala-Arg, N-myristoyl-Ala-Lys, N-myristoyl-Val-His, N-myristoyl-Val-Trp, N-myristoyl-Val-Gln, N-myristoyl-Val-Asn, N-myristoyl-Val-Arg, N-myristoyl-Val-Lys,N-Myristoyl-Leu-His, N-Myristoyl-Leu-Trp, N-Myristoyl-Leu-Gln, N-Myristoyl-Leu-Asn, N-Myristoyl-Leu-Arg, N-Myristoyl-Leu-Lys, N-Myristoyl-Ile-His, N-Myristoyl-Ile-Trp, N-Myristoyl-Ile-Gln, N-Myristoyl-Ile-Asn, N-Myristoyl-Ile-Arg, N-Myristoyl-Ile-Lys, N-Palmitoyl-Gly-His, N-Palmitoyl-Gly-Trp, N-Palmitoyl -Gly-Gln, N-palmitoyl-Gly-Asn, N-palmitoyl-Gly-Arg, N-palmitoyl-Gly-Lys, N-palmitoyl-Ala-His, N-palmitoyl-Ala-Trp, N-palmitoyl-Ala-Gln, N-palmitoyl-Ala-Asn, N-palmitoyl-Ala-Arg, N-palmitoyl-Ala-Lys, N-palmitoyl-Val-His, N-palmitoyl-Val-Trp, N-palmitoyl-Val-Gln, N-palmitoyl-Val-Asn, N-palmitoyl-Val-Arg, N-Palmitoyl-Val-Lys, N-Palmitoyl-Leu-His, N-Palmitoyl-Leu-Trp, N-Palmitoyl-Leu-Gln, N-Palmitoyl-Leu-Asn, N-Palmitoyl-Leu-Arg, N-Palmitoyl-Leu-Lys, N-Palmitoyl-Ile-His, N-Palmitoyl-Ile-Trp, N-Palmitoyl-Ile-Gln, N-Palmitoyl-Ile-Asn, N-Palmitoyl-Ile-Arg, N-Palmitoyl-Ile-Lys, N-Margalloyl-Gly-His, N-Margalloyl -Gly-Trp, N-margalloyl-Gly-Gln, N-margalloyl-Gly-Asn, N-margalloyl-Gly-Arg, N-margalloyl-Gly-Lys, N-margalloyl-Ala-His, N-margalloyl-Ala-Trp, N-margalloyl-Ala-Gln, N-margalloyl-Ala-Asn, N-margalloyl-Ala-Arg, N-margalloyl-Ala-Lys, N-margalloyl-Val-His, N-margalloyl-Val-Trp, N-margalloyl-Val-Gln, N-margalloyl-Val-Asn,N-Margalloyl-Val-Arg, N-Margalloyl-Val-Lys, N-Margalloyl-Leu-His, N-Margalloyl-Leu-Trp, N-Margalloyl-Leu-Gln, N-Margalloyl-Leu-Asn, N-Margalloyl-Leu-Arg, N-Margalloyl-Leu-Lys, N-Margalloyl-Ile-His, N-Margalloyl-Ile-Trp, N-Margalloyl-Ile-Gln, N-Margalloyl-Ile-Asn, N-Margalloyl-Ile-Arg, N-Margalloyl-Ile-Lys, N-Stearoyl -Gly-His, N-Stearoyl-Gly-Trp, N-Stearoyl-Gly-Gln, N-Stearoyl-Gly-Asn, N-Stearoyl-Gly-Arg, N-Stearoyl-Gly-Lys, N-Stearoyl-Ala-His, N-Stearoyl-Ala-Trp, N-Stearoyl-Ala-Gln, N-Stearoyl-Ala-Asn, N-Stearoyl-Ala-Arg, N-Stearoyl-Ala-Lys, N-Stearoyl-Val-His, N-Stearoyl-Val-Trp, N-Stearoyl-Val-Gln, N-Stearoyl-Val-Asn, N-Stearoyl-Val-Arg, N-Stearoyl-Val-Lys, N-Stearoyl-Leu-His, N-Stearoyl-Leu-Trp, N-Stearoyl-Leu-Gln, N-Stearoyl-Leu-Asn, N-Stearoyl-Leu-Arg, N-Stearoyl-Leu-Lys, N-Stearoyl-Ile-His, N-Stearoyl-Ile-Trp, N-Stearoyl-Ile-Gln, N-Stearoyl-Ile-Asn, N-Stearoyl-Ile-Arg, N-Stearoyl -Ile-Lys, N-elidoyl-Gly-His, N-elidoyl-Gly-Trp, N-elidoyl-Gly-Gln, N-elidoyl-Gly-Asn, N-elidoyl-Gly-Arg, N-elidoyl-Gly-Lys, N-elidoyl-Ala-His, N-elidoyl-Ala-Trp, N-elidoyl-Ala-Gln, N-elidoyl-Ala-Asn, N-elidoyl-Ala-Arg, N-elidoyl-Ala-Lys, N-elidoyl-Val-His, N-elidoyl-Val-Trp,N-Elidoyl-Val-Gln, N-Elidoyl-Val-Asn, N-Elidoyl-Val-Arg, N-Elidoyl-Val-Lys, N-Elidoyl-Leu-His, N-Elidoyl-Leu-Trp, N-Elidoyl-Leu-Gln, N-Elidoyl-Leu-Asn, N-Elidoyl-Leu-Arg, N-Elidoyl-Leu-Lys, N-Elidoyl-Ile-His, N-Elidoyl-Ile-Trp, N-Elidoyl-Ile-Gln, N-Elidoyl-Ile-Asn, N-Elidoyl- Ile-Arg, N-Elideyl-Ile-Lys, N-Arachidoyl-Gly-His, N-Arachidoyl-Gly-Trp, N-Arachidoyl-Gly-Gln, N-Arachidoyl-Gly-Asn, N-Arachidoyl-Gly-Arg, N-Arachidoyl-Gly-Lys, N-Arachidoyl-Ala-His, N-Arachidoyl-Ala-Trp, N-Arachidoyl-Ala-Gln, N-Arachidoyl-Ala-Asn, N-Arachidoyl-Ala-Arg, N-Arachidoyl-Ala-Lys, N-Arachidoyl-Val-His, N- Arachidoyl-Val-Trp, N-arachidoyl-Val-Gln, N-arachidoyl-Val-Asn, N-arachidoyl-Val-Arg, N-arachidoyl-Val-Lys, N-arachidoyl-Leu-His, N-arachidoyl-Leu-Trp, N-arachidoyl-Leu-Gln, N-arachidoyl-Leu-Asn, N-arachidoyl-Leu-Arg, N-arachidoyl-Leu-Lys, N-arachidoyl-Ile-His, N-arachidoyl-Ile-Trp, N-arachidoyl-Ile-Gln, N-arachidoyl-Ile e-Asn, N-arachidoyl-Ile-Arg, N-arachidoyl-Ile-Lys, N-behenoyl-Gly-His, N-behenoyl-Gly-Trp, N-behenoyl-Gly-Gln, N-behenoyl-Gly-Asn, N-behenoyl-Gly-Arg, N-behenoyl-Gly-Lys, N-behenoyl-Ala-His, N-behenoyl-Ala-Trp, N-behenoyl-Ala-Gln, N-behenoyl-Ala-Asn, N-behenoyl-Ala-Arg, N-behenoyl-Ala-Lys, N-behenoyl-Val-His,N-Behenoyl-Val-Trp, N-Behenoyl-Val-Gln, N-Behenoyl-Val-Asn, N-Behenoyl-Val-Arg, N-Behenoyl-Val-Lys, N-Behenoyl-Leu-His, N-Behenoyl-Leu-Trp, N-Behenoyl-Leu-Gln, N-Behenoyl-Leu-Asn, N-Behenoyl-Leu-Arg, N-Behenoyl-Leu-Lys, N-Behenoyl-Ile-His, N-Behenoyl-Ile-Trp, N-Behenoyl-Ile-Gln, N-Behenoyl-Ile-Asn, N-Behenoyl-Ile-Arg, N-Behenoyl-Ile-Lys.,

[0019] Among the above compounds, more preferred lipid peptide compounds include N-lauroyl-Gly-His, N-lauroyl-Gly-Trp, N-lauroyl-Gly-Gln, N-lauroyl-Gly-Asn, N-lauroyl-Gly-Lys, N-lauroyl-Ala-His, N-lauroyl-Ala-Trp, N-lauroyl-Ala-Gln, N-lauroyl-Ala-Asn, N-lauroyl-Ala-Lys, N-lauroyl-Val-His, N-lauroyl-Val-Trp, N-lauroyl-Val-Gln, and N-lauroyl-Val-A sn, N-Lauroyl-Val-Lys, N-Myristoyl-Gly-His, N-Myristoyl-Gly-Trp, N-Myristoyl-Gly-Gln, N-Myristoyl-Gly-Asn, N-Myristoyl-Gly-Lys, N-Myristoyl-Ala-His, N-Myristoyl-Ala-Trp, N-Myristoyl-Ala-Gln, N-Myristoyl-Ala-Asn, N-Myristoyl-Ala-Lys, N-Myristoyl-Val-His, N-Myristoyl-Val-Trp, N-Myristoyl-Val-Gln, N-Myristoyl-V al-Asn, N-myristoyl-Val-Lys, N-palmitoyl-Gly-His, N-palmitoyl-Gly-Trp, N-palmitoyl-Gly-Gln, N-palmitoyl-Gly-Asn, N-palmitoyl-Gly-Lys, N-palmitoyl-Ala-His, N-palmitoyl-Ala-Trp, N-palmitoyl-Ala-Gln, N-palmitoyl-Ala-Asn, N-palmitoyl-Ala-Lys, N-palmitoyl-Val-His, N-palmitoyl-Val-Trp, N-palmitoyl-Val-Gln, N-palmitoyl N-palmitoyl-Val-Asn, N-palmitoyl-Val-Lys, N-margalloyl-Gly-His, N-margalloyl-Gly-Trp, N-margalloyl-Gly-Gln, N-margalloyl-Gly-Asn, N-margalloyl-Gly-Lys, N-margalloyl-Ala-His, N-margalloyl-Ala-Trp, N-margalloyl-Ala-Gln, N-margalloyl-Ala-Asn, N-margalloyl-Ala-Lys, N-margalloyl-Val-His, N-margalloyl-Val-Trp, N-margalloyl-Val-Gln,N-Margalloyl-Val-Asn, N-Margalloyl-Val-Lys, N-Stearoyl-Gly-His, N-Stearoyl-Gly-Trp, N-Stearoyl-Gly-Gln, N-Stearoyl-Gly-Asn, N-Stearoyl-Gly-Lys, N-Stearoyl-Ala-His, N-Stearoyl-Ala-Trp, N-Stearoyl-Ala-Gln, N-Stearoyl-Ala-Asn, N-Stearoyl-Ala-Lys, N-Stearoyl-Val-His, N-Stearoyl-Val-Trp, N-Stearoyl- Val-Gln, N-Stearoyl-Val-Asn, N-Stearoyl-Val-Lys, N-Elidoyl-Gly-His, N-Elidoyl-Gly-Trp, N-Elidoyl-Gly-Gln, N-Elidoyl-Gly-Asn, N-Elidoyl-Gly-Lys, N-Elidoyl-Ala-His, N-Elidoyl-Ala-Trp, N-Elidoyl-Ala-Gln, N-Elidoyl-Ala-Asn, N-Elidoyl-Ala-Lys, N-Elidoyl-Val-His, N-Elidoyl-Val-Trp, N- Elaidyl-Val-Gln, N-elaidyl-Val-Asn, N-elaidyl-Val-Lys, N-arachidoyl-Gly-His, N-arachidoyl-Gly-Trp, N-arachidoyl-Gly-Gln, N-arachidoyl-Gly-Asn, N-arachidoyl-Gly-Lys, N-arachidoyl-Ala-His, N-arachidoyl-Ala-Trp, N-arachidoyl-Ala-Gln, N-arachidoyl-Ala-Asn, N-arachidoyl-Ala-Lys, N-arachidoyl-Val-His, N-arachidoyl-Val -Trp, N-arachidoyl-Val-Gln, N-arachidoyl-Val-Asn, N-arachidoyl-Val-Lys, N-behenoyl-Gly-His, N-behenoyl-Gly-Trp, N-behenoyl-Gly-Gln, N-behenoyl-Gly-Asn, N-behenoyl-Gly-Lys, N-behenoyl-Ala-His, N-behenoyl-Ala-Trp, N-behenoyl-Ala-Gln, N-behenoyl-Ala-Asn, N-behenoyl-Ala-Lys, N-behenoyl-Val-His, N-behenoyl-Val-Trp,Examples include N-behenoyl-Val-Gln, N-behenoyl-Val-Asn, and N-behenoyl-Val-Lys.

[0020] The most preferred compounds include N-lauroyl-Gly-His, N-lauroyl-Gly-Gln, N-lauroyl-Gly-Asn, N-lauroyl-Gly-Lys, N-myristoyl-Gly-His, N-myristoyl-Gly-Gln, N-myristoyl-Gly-Asn, N-myristoyl-Gly-Lys, N-palmitoyl-Gly-His, N-palmitoyl-Gly-Trp, N-palmitoyl-Gly-His, N-palmitoyl-Gly-Asn, N-palmitoyl-Gly-Lys, N-palmitoyl-Gly-His, N-palmitoyl-Gly-Trp, N-palmitoyl-Gly-His, N-palmitoyl-Gly-His, N-palmitoyl-Gly-Asn ... N-palmitoyl-Gly-Gln, N-palmitoyl-Gly-Asn, N-palmitoyl-Gly-Lys, N-palmitoyl-Ala-His, N-palmitoyl-Ala-Trp, N-palmitoyl-Ala-Gln, N-palmitoyl-Ala-Asn, N-palmitoyl-Ala-Lys, N-palmitoyl-Val-His, N-palmitoyl-Val-Trp, N-palmitoyl-Val-Gln, N-palmitoyl N-Palmitoyl-Val-Lys, N-Margalloyl-Gly-His, N-Margalloyl-Gly-Gln, N-Margalloyl-Gly-Asn, N-Margalloyl-Gly-Lys, N-Stearoyl-Gly-His, N-Stearoyl-Gly-Gln, N-Stearoyl-Gly-Asn, N-Stearoyl-Gly-Lys, N-Elideyl-Gly-His, N-Elide Examples of such amino acids include N-elidoyl-Gly-Gln, N-elidoyl-Gly-Asn, N-elidoyl-Gly-Lys, N-arachidoyl-Gly-His, N-arachidoyl-Gly-Gln, N-arachidoyl-Gly-Asn, N-arachidoyl-Gly-Lys, N-behenoyl-Gly-His, N-behenoyl-Gly-Gln, N-behenoyl-Gly-Asn, and N-behenoyl-Gly-Lys. Furthermore, examples of lipid peptides having a branched chain structure include N-2-(4,4-dimethylpentan-2-yl)-5,7,7-trimethyloctanoyl-Gly-His and N-2-heptylundecanoyl-Gly-His.

[0021] The base used in the reaction of the ester compound represented by formula (1) with the α-amino acid compound represented by formula (2) is not particularly limited, but examples include alkali metals such as metallic sodium and metallic potassium; alkali metal inorganic acid salts such as sodium carbonate, potassium carbonate, potassium phosphate, and sodium phosphate; alkali metal hydroxides such as sodium hydroxide and potassium hydroxide; alkali metal alkoxides such as sodium methoxide and potassium t-butoxide; aliphatic amines such as triethylamine and tri-n-butylamine; alicyclic amines such as 1,8-diazabicyclo[5.4.0]-7-undecene (hereinafter also referred to as DBU) and 1,5-diazabicyclo[4.3.0]-5-nonene (hereinafter also referred to as DBN); aromatic amines such as pyridine and 2-methyl-5-ethylpyridine; and alcohol solutions or alcohol dispersions of these basic (solid) compounds. These may be used alone or in combination of two or more.

[0022] Among the above bases, in consideration of increasing the conversion rate and further improving the yield of the target product, sodium methoxide, sodium ethoxide, potassium methoxide, potassium ethoxide, potassium t-butoxide, DBU, or DBN is preferred, and an alcohol solution or alcohol dispersion of sodium methoxide or these metal alkoxides is preferred. Sodium methoxide may be in the form of a solid, a methanol solution, or a methanol dispersion. It may also be prepared in advance or in the reaction system using metallic sodium and methanol. Considering operability and yield, it is preferable to use a commercially available approximately 28% methanol solution of sodium methoxide. The amount of base used is not particularly limited and is usually about 1 to 10 equivalents relative to the compound of formula (1), preferably 1 to 5 equivalents, and more preferably 1.3 to 2 equivalents.

[0023] The non-polar organic solvent contained in the solvent used in the above reaction is not particularly limited, and can be appropriately selected from various solvents used in general organic synthesis so long as it does not affect the reaction. Specific examples thereof include saturated aliphatic compounds such as pentane, c-pentane, hexane, c-hexane, methyl c-hexane, heptane, c-heptane, octane, decane, and decalin; unsaturated aliphatic compounds such as 1-hexene and 1-octyne; and aromatic compounds such as benzene, toluene, xylene, and o-dichlorobenzene. These solvents can be used alone or in combination of two or more.

[0024] Among these nonpolar organic solvents, in consideration of preventing hydrolysis of the ester compound represented by formula (1) and increasing the conversion rate to further improve the yield of the target product, at least one selected from the group consisting of toluene, xylene, orthodichlorobenzene, pentane, hexane, heptane, octane, c-pentane, c-hexane, methyl c-hexane, c-heptane, and 1-hexene is preferred, and toluene is particularly suitable.

[0025] The solvent used in the reaction preferably contains an alcohol in addition to the nonpolar solvent. The alcohol used here is not particularly limited, and any alcohol solvent that does not affect the reaction can be appropriately selected from various alcohol solvents used in general organic synthesis. Specific examples thereof include methanol, ethanol, n-propanol, i-propanol, n-butanol, i-butanol, s-butanol, t-butanol, n-pentanol, i-pentanol, s-pentanol, t-pentanol, n-hexanol, i-hexanol, s-hexanol, t-hexanol, octanol, decanol, ethylene glycol, 1,3-butanediol, glycerin, etc. These solvents can be used alone or in combination of two or more.

[0026] The reaction temperature between the ester compound represented by the formula (1) and the α-amino acid compound represented by the formula (2) can be any temperature as long as it is equal to or lower than the boiling point of the solvent used. However, in consideration of obtaining the target product in a short time and with a high yield, the reaction temperature is preferably 20°C to 150°C, more preferably 40°C to 80°C, and even more preferably 65°C to 75°C. The reaction time cannot be generally defined because it varies depending on the reaction temperature, the base used, and the type of organic solvent, but is usually about 1 to 48 hours.

[0027] The reaction can be carried out by mixing all the reagents at room temperature and then heating to the reaction temperature, or by adding the required reagents dropwise to control the reaction. The reaction can also be carried out in a batch or continuous manner, under reduced pressure, normal pressure, or increased pressure. Dropwise addition of the base at normal pressure is more preferred.

[0028] After the reaction is complete, an organic acid is added to neutralize the reaction mixture, and then water and an alcohol are added to remove the non-polar organic solvent by separation. Acetic acid is preferred as the organic acid. In consideration of ease of separation, alkali metal salts of the lipid peptide compound are preferred.

[0029] Thereafter, hydrogen halide is added to the resulting product, preferably in a solvent containing water and alcohol, until the pH reaches the pre-calculated isoelectric point. For example, after the reaction of the ester compound represented by formula (1) with the α-amino acid compound represented by formula (2), the nonpolar organic solvent is removed, and a solution of hydrogen halide is added to the remaining aqueous solution of the lipid peptide compound salt containing alcohol. The isoelectric point, also known as the equipotential point, is the pH value at which the formal charge of the molecule becomes zero in the acid-base dissociation state of the molecule. The isoelectric point value can be calculated from the acid dissociation constant (pKa) of the molecule, and can be calculated from the molecular structure, for example, using Calculator Plugins, a calculation software manufactured by ChemAxon. The isoelectric point can also be calculated from an actually measured zeta potential value.

[0030] The hydrogen halide used in the pH adjustment operation is usually used in the form of an aqueous solution because it is easy to operate, and examples thereof include hydrochloric acid, hydrobromic acid, etc., and preferably hydrochloric acid. Note that when adjusting the pH using hydrogen halide, if the amount required for pH adjustment is exceeded, hydrochloride salts of lipid peptides will be formed, and the recovery rate of the free form will decrease, so care must be taken with the amount of hydrogen halide used.

[0031] After the reaction is completed, the solution is neutralized by adding an organic acid, and the crude lipid peptide compound (free form) is recovered by filtration or the like, and if necessary, subjected to post-treatments such as washing and recrystallization to obtain a purified product.

[0032] The ester compound represented by the above formula (1) used in the present invention can be obtained by reacting a compound represented by the following formula (4) with a compound represented by the following formula (5). [ka] (In the formula, X, R 1 , R 2 and R 3 represents the previously defined

[0033] As described above, in the production method of the present invention, an organic acid is added to the solution after the reaction to neutralize it, and then water and an alcohol are added and the nonpolar organic solvent is removed by a liquid separation operation, so that the nonpolar organic solvent can be easily recovered and disposed of. In addition, when a lipid peptide compound has gelling ability, polar solvents such as DMF that have been used in the production of lipid peptides tend to gel after cooling due to the action of the lipid peptide. However, using a nonpolar organic solvent can prevent gelation, making it very useful in production. Furthermore, although the solution becomes alkaline after the reaction, neutralization can be completed without gelation by using the amount of hydrogen chloride solution required for neutralization, and the free form can be recovered. The precipitated crude crystals of the free form can be purified by known techniques such as recrystallization to obtain the pure target product. On the other hand, when neutralization is not performed, the layer (lower layer) of the alcohol solution of the salt of the lipid peptide compound can be dropped into an organic solvent to reprecipitate and recover the salt of the lipid peptide compound as a solid. [Example]

[0034] The present invention will be explained in more detail below with reference to Synthesis Examples, Examples and Comparative Examples, but the present invention is not limited to the following Examples. The reagents used in the synthesis examples and examples were commercially available as shown below, and the instruments shown below were used for analysis and measurement of physical properties of each synthesized compound.

[0035] Methanol: Kanto Chemical Co., Ltd. (special grade) Tetrahydrofuran: Kanto Chemical Co., Ltd. (1st grade) i-Propanol: Kanto Chemical Co., Ltd. (1st grade) Toluene: Kanto Chemical Co., Ltd. (1st grade) Acetic acid: Kanto Chemical Co., Ltd. (1st grade) Palmitic acid chloride: Aldrich (palmitoyl chloride), NOF Corporation (distilled palmitic acid chloride) Glycine methyl ester hydrochloride: Tokyo Chemical Industry Co., Ltd. Glycine ethyl ester hydrochloride: Tokyo Chemical Industry Co., Ltd. L-histidine: Tokyo Chemical Industry Co., Ltd., Kyowa Hakko Bio Co., Ltd. Sodium methoxide 28% methanol solution: Nippon Soda Co., Ltd. (liquid sodium methylate 28%), Wako Pure Chemical Industries, Ltd. (28% sodium methoxide methanol solution) Sodium carbonate: Junsei Chemical Co., Ltd. (Grade 1), Tokuyama Corporation Hydrochloric acid: Kanto Chemical Co., Ltd. (1st grade) Acetonitrile: Kanto Chemical Co., Ltd. (special grade)

[0036] Amberlist 15JWET: Organo Corporation Amberlite FPC3500: Organo Corporation Kyoword 600: Kyowa Chemical Industry Co., Ltd. Galleon Earth V2: Mizusawa Industrial Chemicals Co., Ltd.

[0037] NMR: JNM-ECP300 (manufactured by JEOL Ltd.) pH meter: Mettler Toledo

[0038] The HPLC analysis conditions are as follows: Column: Inertsil ODS-3 (GL sciences) Developing solvent: MeOH / phosphate buffer (pH=2.1) = 85 / 15 (volume ratio) *Preparation of phosphate buffer solution (pH=2.1) Add 7.8 g (50 mmol) of sodium dihydrogen phosphate (NaH2PO42H2O) and 3.4 mL (50 mmol) of 85% phosphoric acid to water to make a total volume of 1 L. Oven temperature: 40°C Detection method: UV 205 nm Flow rate: 2.0mL / min Injection volume: 20 μL Retention times: N-palmitoyl-Gly-His-methyl...5.0 min, N-palmitoyl-Gly-His...5.5 min, N-palmitoyl-Gly...9.3 min, N-palmitoyl-Gly-methyl...11.2 min

[0039] [Example 1] Synthesis of N-palmitoyl-Gly-ethyl A 500 mL four-neck flask was charged with 15.2 g (0.11 mol) of glycine ethyl ester hydrochloride and 50 g of water. Then, 10.6 g (0.10 mol) of sodium carbonate (base), 75 g of water, and 100 g of toluene (organic solvent) were added and stirred. Then, 25.0 g (0.090 mol) of palmitic acid chloride was added dropwise over 1 hour at a reaction temperature of 40-45°C. After stirring for 2 hours, 75 g of 10% saline was added and the mixture was separated at 60°C. To the resulting organic layer, 200 g of toluene was added and azeotropic dehydration was performed to obtain 248.0 g of a toluene solution of N-palmitoyl-Gly-ethyl (100% yield).

[0040] · 1H-NMR(300MHz,CDCl3,δppm):5.94(1H,m),2.18(2H,q,J=7.5Hz),4.03(2H,d,J= 5.4Hz),2.24(2H,t,J=7.2Hz),1.62(4H,m),1.31(25H,m),0.88(3H,t,J=7.2Hz) MS(CI) m / z: 342.10(M + +1) Melting point: 79.5℃

[0041] [Example 2] A 1-L four-neck flask was charged with 14.1 g (0.090 mol) of histidine and 62.1 g of toluene, and 16.7 g (0.086 mol) of a 28% methanol solution of sodium methoxide (a base) was added dropwise. The toluene solution of N-palmitoyl-Gly-ethyl obtained in Example 1 was azeotropically dehydrated, and then added together with 12.4 g of methanol. The mixture was heated to 70°C. Then, 13.2 g (0.068 mol) of a 28% methanol solution of sodium methoxide (a base) was added dropwise, and stirring was continued at approximately 70°C for 3 hours. After the reaction was completed, the mixture was cooled to 55°C, the pH was adjusted to 7 using acetic acid, and 155.3 g of water and 62.1 g of 2-propanol were added to separate the liquids. The lower layer was removed, mixed with 869.8 g of water, and the mixture was adjusted to 40°C. The mixture was then neutralized with 35% hydrochloric acid to pH 4.5, resulting in the precipitation of crude crystals of N-palmitoyl-Gly-His· free form. After cooling, the crystals were filtered and dried under reduced pressure at 80°C, yielding 36.5 g of crude crystals. To the resulting solid, 900 g of water and 1,800 g of methanol were added and the mixture was heated and stirred at 60°C for 1 hour. The mixture was then allowed to cool to 25°C, and the precipitated solid was collected by filtration. The same procedure was repeated once more, and the resulting solid was dried under reduced pressure. Next, 650 g of tetrahydrofuran was added to the dried solid, and the mixture was stirred at 25°C for 1 hour. The solid was then collected by filtration. 1,300 g of methanol and 650 g of tetrahydrofuran were added to the resulting solid, and the mixture was heated to 60°C and dissolved. The mixture was then cooled to 0°C over 2 hours and stirred at 0°C overnight. The precipitated solid was collected by filtration and dried under reduced pressure, yielding 31.3 g of white crystals of N-palmitoyl-Gly-His· (free form) (purity: 100%, yield: 91.1%).

[0042] [Comparative Example 1] After the reaction in Example 2 was completed, the mixture was cooled to 60°C, and 12.7 g of ion exchange resin (Amberlite FPC3500) was added, changing the pH from 11 to pH 7. The ion exchange resin was filtered, and the resulting solution was reprecipitated in 135.0 g of acetonitrile to obtain free N-palmitoyl-Gly-His. HPLC analysis revealed that a methylated compound of N-palmitoyl-Gly-His was produced at an area percentage of 0.2%.

[0043] Comparative Example 2 Free N-palmitoyl-Gly-His· was obtained in the same manner as in Comparative Example 1, except that the ion exchange resin used was changed (Amberlyst 15JWET 13.3 g). HPLC analysis revealed that a methylated compound of N-palmitoyl-Gly-His was produced at an area percentage of 5.6%.

[0044] Comparative Example 3 The same procedure was carried out as in Example 3, except that the ion exchange resin was changed to a clay mineral (Galleon Earth V2 10.0 g), but the pH simply changed from 11 to 10 and neutralization was not complete.

[0045] Comparative Example 4 The same procedure was carried out as in Example 3, except that the ion exchange resin was changed to magnesium silicate (Kyoward 600 10.0 g), but the pH simply changed from 11 to 10 and neutralization was not complete.

[0046] [Example 3] Synthesis of N-palmitoyl-Gly-ethyl A 500L reactor was charged with 5.48 kg (39.3 mol) of glycine ethyl ester hydrochloride and 18 kg of water. 3.82 kg (36.0 mol) of sodium carbonate (base), 27 kg of water, and 36 kg of toluene (organic solvent) were added and stirred. 9.0 kg (32.7 mol) of palmitic acid chloride was then added dropwise over 1 hour at a reaction temperature of 40-45°C. After stirring for 2 hours, 27 kg of 10% brine was added and the mixture was separated at 60°C. 59.6 kg of toluene was added to the resulting organic layer, and azeotropic dehydration was performed to obtain 89.5 kg of a toluene solution of N-palmitoyl-Gly-ethyl (100% yield).

[0047] [Example 4] A 200 L reaction vessel was charged with 5.08 kg (32.7 mol) of histidine and 22.4 kg of toluene, and 6.00 kg (31.1 mol) of a 28% methanol solution of sodium methoxide (base) was added dropwise. The toluene solution of N-palmitoyl-Gly-ethyl obtained in Example 3 was then added together with 4.47 kg of methanol, and the mixture was heated to 70 °C. Subsequently, dropwise addition of 4.74 kg (24.6 mol) of a 28% methanol solution of sodium methoxide (base) was initiated, and stirring was continued at approximately 70 °C for 4 hours. After the reaction was completed, the mixture was cooled to 55 °C, the pH was adjusted to 7.6 using acetic acid, and 55.9 kg of water and 22.4 kg of 2-propanol were added to separate the mixture. The lower layer was removed, mixed with 313.1 kg of water, and the mixture was adjusted to 40°C. The mixture was then neutralized with 35% hydrochloric acid to pH 4.5, resulting in the precipitation of crude crystals of N-palmitoyl-Gly-His· free form. After cooling, the crystals were filtered and dried under reduced pressure at 80°C, yielding 36.5 kg of crude crystals. Recrystallization was carried out in a 500 L reaction vessel in the same manner as in Example 2, yielding 12.0 kg of white crystals of N-palmitoyl-Gly-His· free form (purity 100%, yield 91.1%).

Claims

1. Formula (1) 【Chemistry 1】 (In the formula, R １ represents an aliphatic group having a linear or branched chain structure and having 9 to 23 carbon atoms; R ２ represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms, which may have a branched chain having 1 or 2 carbon atoms; R ３ represents an alkyl group having 1 to 6 carbon atoms, a haloalkyl group having 1 to 6 carbon atoms, a hydroxyalkyl group having 1 to 6 carbon atoms, or an aryl group which may be substituted with an alkyl group having 1 to 6 carbon atoms. 【Chemistry 2】 (In the formula, R ４ Ha-(CH ２ ) ｎ represents an —X group, n is a number from 1 to 4, and X is an amino group, a guanidino group, or —CONH ２ a reaction step of reacting an α-amino acid compound represented by the formula (I) with a base in a solvent containing a non-polar organic solvent; The compound of formula (3) obtained in this reaction step 【Transformation 3】 (In the formula, R １ , R ２ and R ４ represents the above-defined meaning.) An extraction step of adding an organic acid to a solution in which a salt of a lipid peptide represented by the formula (I) is dissolved, neutralizing the solution, then adding water and alcohol to separate the solution, and removing the non-polar organic solvent; A method for producing a lipid peptide compound represented by formula (3) or a pharmaceutically usable salt thereof, comprising a separation step of removing the lipid peptide compound represented by formula (3) from the solution after the extraction step.

2. The method according to claim 1 , wherein the solvent comprises a non-polar organic solvent and an alcohol.

3. In the formula, n represents a number from 1 to 4, and X represents an amino group, a guanidino group, or -CONH ２ 2. The method according to claim 1, wherein n represents a group, or n represents 1 and X represents a pyrrole group, an imidazole group, a pyrazole group or an imidazole group.

4. In the formula, R １ The method according to claim 1, wherein represents a linear aliphatic group having 11 to 21 carbon atoms and which may have 0 to 2 unsaturated bonds.

5. In the formula, R ２ 2. The process according to claim 1, wherein represents a hydrogen atom or an alkyl group having 1 to 3 carbon atoms, which may have a branched chain containing 1 carbon atom.

6. In the formula, R ２ represents a hydrogen atom, a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a sec-butyl group, or a tert-butyl group; R ４ represents an aminomethyl group, an aminoethyl group, a 3-aminopropyl group, a 4-aminobutyl group, a carbamoylmethyl group, a 2-carbamoylethyl group, a 3-carbamoylbutyl group, a 2-guanidinoethyl group, a 3-guanidinopropyl group, a pyrrolemethyl group, an imidazolemethyl group, a pyrazolemethyl group, or a 3-indolemethyl group.

7. In the formula, R ２ represents a hydrogen atom, a methyl group, an isopropyl group, an isobutyl group, or a sec-butyl group; R ４ The method according to claim 6, wherein represents a 4-aminobutyl group, a carbamoylmethyl group, a 2-carbamoylethyl group, a 3-guanidinopropyl group, an imidazolemethyl group, or a 3-indolemethyl group.

8. The method according to claim 1 , wherein the organic acid is acetic acid.

9. 9. The method according to claim 1, wherein the base is at least one selected from the group consisting of alkali metals, inorganic alkali metal salts, alkali metal hydroxides, alkali metal alkoxides, alicyclic amines, alcohol solutions thereof, and alcohol dispersions thereof.

10. The production method according to claim 9, wherein the base is at least one selected from metallic sodium, metallic potassium, sodium carbonate, potassium carbonate, potassium phosphate, sodium phosphate, sodium hydroxide, potassium hydroxide, sodium methoxide, sodium ethoxide, potassium methoxide, potassium ethoxide, potassium t-butoxide, 1,8-diazabicyclo[5.4.0]-7-undecene, 1,5-diazabicyclo[4.3.0]-5-nonene, an alcohol solution thereof, or an alcohol dispersion thereof.

11. The method according to claim 10, wherein the base is sodium methoxide, a methanol solution thereof, or a methanol dispersion thereof.

12. 12. The method according to claim 1, wherein the non-polar organic solvent is at least one selected from the group consisting of aromatic compounds, saturated aliphatic compounds, and unsaturated aliphatic compounds.

13. The production method according to claim 12, wherein the nonpolar organic solvent is at least one selected from the group consisting of toluene, xylene, orthodichlorobenzene, pentane, hexane, heptane, octane, cyclopentane, cyclohexane, methylcyclohexane, cycloheptane, and 1-hexene.

14. The method of claim 2 , wherein the solvent comprises toluene and methanol or ethanol.

15. The production method according to any one of claims 1 to 14, wherein the reaction of the ester compound represented by formula (1) with the α-amino acid compound represented by formula (2) is carried out at a reaction temperature of 65°C to 75°C.

16. Formula (1) 【Chemistry 4】 (In the formula, R １ represents an aliphatic group having a linear or branched chain structure and having 9 to 23 carbon atoms; R ２ represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms, which may have a branched chain having 1 or 2 carbon atoms; R ３ represents an alkyl group having 1 to 6 carbon atoms, a haloalkyl group having 1 to 6 carbon atoms, a hydroxyalkyl group having 1 to 6 carbon atoms, or an aryl group which may be substituted with an alkyl group having 1 to 6 carbon atoms. 【Transformation 5】 (In the formula, R ４ Ha-(CH ２ ) ｎ represents an —X group, n is a number from 1 to 4, and X is an amino group, a guanidino group, or —CONH ２ a reaction step of reacting an α-amino acid compound represented by the formula (I) with a base in a solvent containing a non-polar organic solvent; The compound of formula (3) obtained in this reaction step 【Transformation 6】 (In the formula, R １ , R ２ and R ４ represents the above-defined meaning.) An extraction step in which an organic acid is added to a solution in which a salt of the lipid peptide represented by the formula (I) is dissolved to neutralize the solution, and then water and alcohol are added to separate the solution, and the non-polar organic solvent is removed; a pH adjustment step of adjusting the pH of the solution after the extraction step using hydrogen halide; and A method for producing a lipid peptide compound represented by formula (3), comprising a separation step of removing the lipid peptide compound represented by formula (3) from the solution after the pH adjustment step.

17. Formula (4) 【Transformation 7】 (wherein X is a halogen atom, an alkoxy group having 1 to 6 carbon atoms, or —OC(O)R １ represents a group, and R １ represents an aliphatic group having a linear or branched chain structure and having 9 to 23 carbon atoms, and a compound represented by formula (5): 【Transformation 8】 (In the formula, R ２ represents a hydrogen atom or an alkyl group having 1 to 4 carbon atoms, which may have a branched chain having 1 or 2 carbon atoms; R ３ represents an alkyl group having 1 to 6 carbon atoms, a haloalkyl group having 1 to 6 carbon atoms, a hydroxyalkyl group having 1 to 6 carbon atoms, or an aryl group which may be substituted with an alkyl group having 1 to 6 carbon atoms. 【Chemistry 9】 (In the formula, R １ , R ２ and R ３ represents the same as defined above, The ester compound represented by formula (1) obtained in the production step and the ester compound represented by formula (2) 【Chemistry 10】 (In the formula, R ４ is a hydrogen atom, an alkyl group having 1 to 7 carbon atoms which may have a branched chain and which has 1 to 3 carbon atoms, a phenylmethyl group, a phenylethyl group, -(CH ２ ) ｎ represents an —X group, n is a number from 1 to 4, and X is an amino group, a guanidino group, or —CONH ２ a reaction step of reacting an α-amino acid compound represented by the formula (I) with a base in a solvent containing a non-polar organic solvent; The compound of formula (3) obtained in this reaction step 【Chemistry 11】 (In the formula, R １ , R ２ and R ４ represents the above-defined meaning.) An extraction step of adding an organic acid to a solution in which a salt of a lipid peptide represented by the formula (I) is dissolved, neutralizing the solution, then adding water and alcohol to separate the solution, and removing the non-polar organic solvent; A method for producing a lipid peptide compound represented by formula (3) or a pharmaceutically usable salt thereof, comprising a separation step of removing the lipid peptide compound represented by formula (3) from the solution after the extraction step.

18. The method according to claim 17, wherein the reaction of the compound represented by formula (4) with the compound represented by formula (5) is carried out in a homogeneous phase at a reaction temperature of 35°C to 45°C.

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