Method for synthesizing cyclic depsipeptides

Abstract

The present invention relates to a method for synthesizing cyclic depsipeptides, particularly emodepsides, from their open forms.

Description

[Technical Field]

[0001] The present invention relates to a method for synthesizing cyclic depsipeptides, particularly emodepsides, that include specific carboxylic acid group protection tags. [Background technology]

[0002] Emodepside (cyclo[(R)-lactoyl-N-methyl-L-leucyl-(R)-3-(p-morpholinophenyl)lactoyl-N-methyl-L-leucyl-(R)-lactoyl-N-methyl-L-leucyl-(R)-3-(p-morpholinophenyl)lactoyl-N-methyl-L-leucyl) is an effective anthelmintic against several gastrointestinal nematodes. Its molecular structure, shown below, can be described as a cyclic octadepsipeptide, a depsipeptide being a peptide in which one or more of its amide groups are substituted with corresponding ester groups. On a technical scale, emodepside can be obtained by derivatization of the natural substance PF1022A in which two hydrogen atoms are replaced by a morpholine ring. [ka]

[0003] International Publication No. 93 / 19053 (European Patent No. 0634408 specification) is a general formula: [ka] (wherein A is a benzyl group having one or more suitable substituents or a phenyl group that may have one or more suitable substituents, A a (where is a benzyl group that may have one or more suitable substituents, or a phenyl group that may have one or more suitable substituents, B and D are each lower alkyl groups, and C is hydrogen or a lower alkyl group) This document discloses compounds and their pharmaceutically acceptable salts.

[0004] International Publication No. 2005 / 055973 discloses transdermal agents containing cyclic depsipeptides and / or praziquantel, in addition to their manufacture and use in combating endoparasites. Combinations of emodepside with praziquantel or epsiplantel, and also with 1,2-isopropylidene glycol, as endoparasiticides are disclosed in International Publication No. 2006 / 094664.

[0005] International Publication No. 2006 / 053641 concerns the use of endoparasitic depsipeptides for the manufacture of pharmaceuticals to prevent vertical infections caused by endoparasites.

[0006] In particular, the total synthesis and derivatization of PF1022A are discussed in the review article "Cyclodepsipeptides: A Rich Source of Biologically Active Compounds for Drug Research" by Sivatharushan Sivanathan and Jurgen Scherkenbeck, Molecules 2014, 19, 12368~12420; doi:10.3390 / molecules190812368. Total synthesis of several cyclodepsipeptides has been established in both solution and solid phase, enabling the preparation of combinatorial libraries. Furthermore, the biosynthesis of certain cyclodepsipeptides has been elucidated and is used for the chemienzymatic preparation of unnatural analogs. The review article also summarizes recent literature on cyclic tetra-~decadepsipeptides composed solely of α-amino acids and α-hydroxy acids.

[0007] In polypeptide synthesis, solid-phase strategies have the advantage of easy separation of reaction products, while liquid-phase strategies have the advantage of homogeneous reaction conditions. The hybrid approach is a tag-assisted strategy in which compounds with tagged groups are easily separated from untagged molecules.

[0008] In this regard, Japanese Patent No. 2000 / 044493 (republished) discloses a protecting group for synthesizing a compound library consisting of compounds that can be bound in a 1:1 ratio to a protected compound, have a single molecular structure, and have a molecular weight >= 500, comprising 3,4,5-tris-(n-octadecyloxy)benzyl alcohol, 3,4,5-tris-(n-octadecyloxy)benzyl chloride, or methyl 3,4,5-tris-(n-octadecyloxy)benzoate.

[0009] European Patent No. 2003104 relates to an organic synthesis reagent that allows chemical reactions to be carried out in the liquid phase and enables the easy and low-cost separation of one or more unwanted compounds from the liquid phase after the reaction is complete. The organic synthesis reagent described below has been reported to reversibly change from a liquid state to a solid state by changes in solution composition and / or solution temperature, and is intended for use in organic synthesis reactions. [ka]

[0010] R1 to R5 may be the same or different, and represent hydrogen, halogen, optionally substituted C1 to C30 alkyl group, optionally substituted C1 to C30 alkoxyl group, optionally substituted C1 to C30 aryl group, optionally substituted C1 to C30 acyl group, optionally substituted C1 to C30 thioalkyl group, optionally substituted C1 to C30 dialkylamino group, nitro group or amino group; at least two of R1 to R5 are C18 to C30 groups, and X represents a reagent active site having one or more atoms selected from the group consisting of carbon atoms, oxygen atoms, sulfur atoms and nitrogen atoms.

[0011] The publication "Tag-Assisted Liquid-Phase Peptide Synthesis Using Hydrophobic Benzyl Alcohols as Supports" by Yohei Okada, Hideaki Suzuki, Takashi Nakae, Shuji Fujita, Hitoshi Abe, Kazuo Nagano, Toshihide Yamada, Nobuyoshi Ebata, Shokaku Kim, and Kazuhiro Chiba, The Journal of Organic Chemistry 2013, 78, 320-327; doi:10.1021 / jo302127d reports the successful establishment of soluble tag-assisted liquid-phase peptide synthesis based on simple hydrophobic benzyl alcohols readily available from naturally abundant materials. It is reported that excellent precipitation yields are obtained at each step by combining the best characteristics of solid-phase and liquid-phase techniques. This approach is reported to be efficiently applicable to fragment coupling and enables the chemical synthesis of several bioactive peptides.

[0012] The publication "A Novel Protecting Group for Constructing Combinatorial Peptide Libraries" by Hitoshi Tamiaki, Tomoyuki Obata, Yasuo Azefu, and Kazunori Toma, Bulletin of the Chemical Society of Japan 2001, 74, 733-738; doi http: / / dx.doi.org / 10.1246 / bcsj.74.733 describes 3,4,5-tris(octadecyloxy)benzyl alcohol, prepared from gallic acid and stearyl bromide, HO-Bzl(OC 18 )3 is disclosed. Using conventional stepwise extension, N,C protective peptide, Fmoc-AA n -...-AA1-OBzl(OC 18 )3 was synthesized. The substituted benzyl ester was selectively cleaved by treatment with 4M hydrogen chloride in ethyl acetate, resulting in Fmoc-AAn -...-AA1-OH and HO-Bzl(OC 18 )3 was obtained. Therefore, it has been reported that the substituted benzyl group is effective for protecting the C-terminal carboxyl group in liquid-phase peptide synthesis. Since the substituted benzyl group has a moderately high molecular weight, Fmoc-AA n -...-AA1-OBzl(OC 18 )3 has been reported to be easily purified by size-exclusion chromatography; all of the protected peptides were reported to elute in the void fraction of a Sephadex LH-20 gel filtration column. The combination of the carboxyl protecting group Bzl(OC 18 )3 and simple purification by gel filtration has been reported to provide a novel route for constructing combinatorial peptide libraries in solution phase.

[0013] International Publication No. 2017 / 116702 pamphlet relates to a cyclic depsipeptide compound of the following formula or a pharmaceutically or veterinarily acceptable salt:

Chemical formula

Chemical formula

Prior art documents

Patent documents

[0014]

Patent document 1

Patent document 2

Patent document 3

[0015] [Non-Patent Document 1] "Cyclodepsipeptides: A Rich Source of Biologically Active Compounds for Drug Research" Sivatharushan Sivanathan and Jurgen Scherkenbeck, Molecules 2014, 19, 12368~12420;doi:10.3390 / molecules190812368 [Non-Patent Document 2] "Tag-Assisted Liquid-Phase Peptide Synthesis Using Hydrophobic Benzyl Alcohols as Supports" Yohei Okada, Hideaki Suzuki, Takashi Nakae, Shuji Fujita, Hitoshi Abe, Kazuo Nagano, Toshihide Yamada, Nobuyoshi Ebata, Shokaku Kim and Kazuhiro Chiba, The Journal of Organic Chemistry 2013, 78, 320~327;doi:10.1021 / jo302127d [Non-Patent Document 3] "A Novel Protecting Group for Constructing Combinatorial Peptide Libraries" Hitoshi Tamiaki, Tomoyuki Obata, Yasuo Azefu and Kazunori Toma, Bulletin of the Chemical Society of Japan 2001, 74, 733~738;doi http: / / dx.doi.org / 10.1246 / bcsj.74.733 [Overview of the Initiative] [Problems that the invention aims to solve]

[0016] The present invention aims to provide an improved synthetic route to emodepsides and related cyclic depsipeptides. Another object of the present invention is to provide novel depsipeptides that can be synthesized using this route. [Means for solving the problem]

[0017] This objective is achieved by the method described in claim 1 and by the depsipeptide described in claim 22. Preferred embodiments are the subject of the dependent claims. These can be freely combined unless explicitly indicated in the context.

[0018] Therefore, the present invention provides a method for synthesizing a cyclic depsipeptide according to general formula (I) from a depsipeptide according to general formula (II): [ka] (In the formula, B is an amine protecting group and A is a carboxylic acid protecting group.) And, - The step of deprotecting the amine group protected by the B group, thereby obtaining the deprotected amine group; - The step of deprotecting a carboxylic acid protected by an A group, thereby obtaining a deprotected carboxylic acid group; - A step of condensing a deprotected amine group with a carboxylic acid group to obtain a cyclic depsipeptide (I) and Includes, x and y are independently 0, 1, or 2, where x + y ≥ 1 (preferably x and y are 1), and R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, and R12 are each independently hydrogen, linear or branched C1-C8 alkyl, particularly methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, sec-pentyl, hexyl, isohexyl, sec-hexyl, heptyl, isoheptyl, sec-heptyl, tert- Heptyl, octyl, isooctyl, sec-octyl, linear or branched halogenated C1-C8 alkyl, especially sec-butyl fluoride, hydroxy-C1-C6 alkyl, especially hydroxymethyl, 1-hydroxyethyl, C1-C4-alkanoyloxy-C1-C6 alkyl, especially acetoxymethyl, 1-acetoxyethyl, C1-C4-alkoxy-C1-C6 alkyl, especially methoxymethyl, 1-methoxyethyl, aryl-C1-C4-alkyloxy-C1-C6 alkyl, especially benzyloxymethyl, 1-benzyloxyethyl L, mercapto-C1~C6-alkyl, especially mercaptomethyl, C1~C4-alkylthio-C1~C6-alkyl, especially methylthioethyl, C1~C4-alkylsulfinyl-C1~C6-alkyl, especially methylsulfinylethyl, C1~C4-alkylsulfonyl-C1~C6-alkyl, especially methylsulfonylethyl, carboxy-C1~C6-alkyl, especially carboxymethyl, carboxyethyl, C1~C4-alkoxycarbonyl-C1~C6-alkyl, especially methoxycarbonylmethyl, ethoxycarbonylethyl, C1~C 4-arylalkoxycarbonyl-C1~C6-alkyl, especially benzyloxycarbonylmethyl, carbamoyl-C1~C6-alkyl, especially carbamoylmethyl, carbamoylethyl, amino-C1~C6-alkyl, especially aminopropyl, aminobutyl, C1~C4-alkylamino-C1~C6-alkyl, especially methylaminopropyl, methylaminobutyl, C1~C4-dialkylamino-C1~C6-alkyl, especially dimethylaminopropyl, dimethylaminobutyl, guanidino-C1~C6-alkyl, especially guanidinopropyl,C1-C4 alkoxycarbonylamino-C1-C6 alkyl, especially tert-butoxycarbonylaminopropyl, tert-butoxycarbonylaminobutyl, 9-fluorenylmethoxycarbonyl-(Fmoc)amino-C1-C6 alkyl, especially 9-fluorenylmethoxycarbonyl(Fmoc)aminopropyl, 9-fluorenylmethoxycarbonyl-(Fmoc)-aminobutyl, C2-C8 alkenyl, especially vinyl, allyl, butenyl, C3-C7 cycloalkyl, especially cyclo Lopentyl, cyclohexyl, cycloheptyl, C3-C7-cycloalkyl-C1-C4-alkyl, especially cyclopentylmethyl, cyclohexylmethyl, cycloheptylmethyl, benzyl, substituted benzyl, phenyl, phenyl-C1-C4-alkyl, especially phenyl-methyl which may be substituted with a group from the group consisting of halogens, especially fluorine, chlorine, bromine or iodine, hydroxyl, C1-C4-alkoxy, especially methoxy or ethoxy, C1-C4-alkyl, especially methyl Provide a method.

[0019] Protecting groups A and B may be orthogonal such that group A is stable when group B is deprotected; however, it is also an embodiment of the present invention that A and B are deprotected simultaneously.

[0020] Surprisingly, the method according to the present invention has been found to enable the synthesis of cyclic depsipeptides (I), particularly emodepsides and closely related structures, in high overall yield.

[0021] After deprotection, the amine group and carboxylic acid group react to form a peptide bond. Suitable coupling agents when combined with a base such as N,N-diisopropylethylamine include PyBOP (benzotriazole-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate), BOP ((benzotriazole-1-yloxy)-tris-(dimethylamino)phosphonium hexafluorophosphate), BOP-Cl (bis(2-oxo-3-oxazolidinyl)phosphinate chloride), EDCI (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide), DEPBT (3-(diethoxyphosphoryloxy)-1, This includes 2,3-benzotriazine-4(3H)-one), HATU (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate), HBTU (2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate), and most preferably T3P(registered trademark) (propylphosphonic anhydride, 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphospholinane-2,4,6-trioxide, PPACA).

[0022] In one embodiment of the method according to the present invention, x is 1, y is 1, R1, R4, R7 and R10 are methyl, R6 and R12 are methyl, R5 and R11 are independently linear or branched C1-C4 alkyl or linear or branched halogenated C1-C4 alkyl, and R3 and R9 are independently benzyl or substituted benzyl. According to one preferred embodiment of the invention, R3 and / or R9 is p-morpholino-substituted benzyl.

[0023] According to embodiments of the present invention, A is acid-unstable, and B is unstable to hydrogenolysis. Therefore, a method for synthesizing a cyclic depsipeptide according to general formula (I) from a depsipeptide according to general formula (IIa): [ka] (In the formula, Y is an amine protecting group and X is a carboxylic acid protecting group.) And, - The step of deprotecting an amine group protected by a Y group in the presence of an acid, thereby obtaining a deprotected amine group; -The step of deprotecting the carboxylic acid protected by the X group via hydrocracking, thereby obtaining the deprotected carboxylic acid group; - A step of condensing a deprotected amine group with a carboxylic acid group to obtain a cyclic depsipeptide (I) and One embodiment of the present invention is to provide a method that includes the following.

[0024] Surprisingly, the method according to this embodiment has been found to enable the synthesis of cyclic depsipeptides (I), particularly emodepsides and closely related structures, in high overall yield.

[0025] The term "hydrocracking" should be understood in its broadest sense and is not explicitly limited to reactions with gaseous and / or molecular hydrogen, although this is one embodiment of the present invention. Suitable catalysts are Pd, Pd / C, Pt, and Pt / C. The term "hydrocracking" also means reactions in which hydrogen is formed in situ or formally only, and in which hydrocracking reactants such as hydrazine or diimide are used.

[0026] In one embodiment, X is a substituted or unsubstituted CH2-aryl group. According to one embodiment, X is selected from the group consisting of benzoyl (Bn), 4-methoxybenzoyl (PMB), 3,4-dimethoxybenzoyl (DPMB), 4-phenylbenzoyl (PPB), 2-naphthylmethyl (Nap), and benzyloxymethyl acetal (BOM).

[0027] In one embodiment of the present invention, Y is t-butyloxycarbonyl (Boc), trityl (Trt), p-methoxybenzylcarbamate (Moz), or p-nitrobenzylcarbamate (PNZ). The most preferred Y is Boc.

[0028] In another embodiment of the method according to the present invention, a depsipeptide according to general formula (IIa) is -In precursor (IV), the amine group protected by the PG2 group is deprotected in the presence of a base, thereby obtaining the deprotected amine group; -In precursor (III), the carboxylic acid protected by the PG3 group is deprotected in the presence of an acid, thereby obtaining the deprotected carboxylic acid group; - The deprotected amine group and carboxylic acid group are condensed to obtain a depsipeptide (IIa). By doing so, Obtained from precursors according to general formulas (IV) and (III): [ka] (In the formulas, R1-R12, X, Y, x, and y have the meanings defined above (in particular, x and y may be 1), PG2 is an amine protecting group, and PG3 is a carboxylic acid protecting group). The deprotection and condensation methods may be the same as those outlined in relation to the reactions of compounds (IIa) through (I) above.

[0029] Preferably, the precursor according to general formula (IV) is -In precursor (VI), the amine group protected by the PG4 group is deprotected in the presence of a base, thereby obtaining the deprotected amine group; -The deprotected amine group of precursor (VI) and the carboxylic acid group of precursor (V) are condensed to obtain precursor (IV). By doing so, Obtained from precursors according to general formulas (VI) and (V): [ka] (In the formulas, R7-R12, X and x have the meanings defined above (in particular, x may be 1), PG2 has the meaning defined above, and PG4 is an amine protecting group). The deprotection and condensation methods may be the same as those outlined in relation to the reaction of compounds (II) to (I) above.

[0030] The precursor according to general formula (III) is -In precursor (VII), the amine group protected by the PG5 group is deprotected in the presence of a base, thereby obtaining the deprotected amine group; -The deprotected amine group of precursor (VII) is condensed with the carboxylic acid group of precursor (VIII) to obtain precursor (III). By doing so, Obtained from precursors according to general formulas (VIII) and (VII): [ka] (In the formula, R1 to R6, Y and y have the meanings defined above (in particular, y may be 1), PG3 has the meaning defined above, and PG5 is an amine protecting group.) It is also preferable that the deprotection and condensation methods be the same as those outlined in relation to the reaction of compounds (II) to (I) described above.

[0031] The precursor according to general formula (VI) is obtained by esterification of the precursor according to general formula (IX) with X-LG: [ka] (In the formula, R10~R12 and X have the meanings defined above, PG4 also has the meaning defined above, and LG is a leaving group.) It is also preferable that the carboxylic acid group of (IX) be protected or tagged using standard procedures, and LG is usually a halide, especially a chloride. Alternatively, LG may be -OH, in which case a standard condensation protocol is usually applied.

[0032] The precursor according to general formula (VII) is obtained by esterification of the precursor according to general formula (X) with PG3-OH: [ka] (In the formula, R4~R6 and y have the meanings defined above, PG3 also has the meanings defined above, and PG5 also has the meanings defined above.) That is also desirable.

[0033] It is also preferable that precursors (III) and (IV) are the same.

[0034] It is also preferable that R3 and R9 are the same, R1 and R7 are the same, R2 and R8 are the same, R4 and R10 are the same, R5 and R11 are the same, and R6 and R12 are the same.

[0035] According to an alternative embodiment of the present invention, A is base-unstable and B is acid-unstable. Therefore, an alternative embodiment of the present invention is a method for synthesizing a cyclic depsipeptide according to general formula (I) from a depsipeptide according to general formula (IIb): [ka] (In the formula, PG1 is an amine protecting group, and TAG is a carboxylic acid protecting group.) And, - The step of deprotecting an amine group protected by a PG1 group in the presence of a base, thereby obtaining a deprotected amine group; - The step of deprotecting a carboxylic acid protected by a TAG group in the presence of an acid, thereby obtaining a deprotected carboxylic acid group; - A step of condensing a deprotected amine group with a carboxylic acid group to obtain a cyclic depsipeptide (I) and This provides a method that includes [something].

[0036] The TAG group is a partial Aryl-O-(CH2) group. n- (Aryl represents the aromatic moiety, and n is 13 or greater), x and y are independently 0, 1 or 2, where x + y ≥ 1 (preferably x and y are 1), and R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11 and R12 are each independently hydrogen, linear or branched C1-C8 alkyl, particularly methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, sec-pentyl, hexyl, isohexyl, sec-hexyl Xyl, heptyl, isoheptyl, sec-heptyl, tert-heptyl, octyl, isooctyl, sec-octyl, linear or branched halogenated C1-C8 alkyl, especially sec-butyl fluoride, hydroxy-C1-C6 alkyl, especially hydroxymethyl, 1-hydroxyethyl, C1-C4-alkanoyloxy-C1-C6 alkyl, especially acetoxymethyl, 1-acetoxyethyl, C1-C4-alkoxy-C1-C6 alkyl, especially methoxymethyl, 1-methoxyethyl, aryl-C1-C4-alkyloxy-C1-C6 -Alkyl, especially benzyloxymethyl, 1-benzyloxyethyl, mercapto-C1~C6-alkyl, especially mercaptomethyl, C1~C4-alkylthio-C1~C6-alkyl, especially methylthioethyl, C1~C4-alkylsulfinyl-C1~C6-alkyl, especially methylsulfinylethyl, C1~C4-alkylsulfonyl-C1~C6-alkyl, especially methylsulfonylethyl, carboxy-C1~C6-alkyl, especially carboxymethyl, carboxyethyl, C1~C4-alkoxycarbonyl-C1~C6-alkyl, especially Methoxycarbonylmethyl, ethoxycarbonylethyl, C1-C4-arylalkoxycarbonyl-C1-C6-alkyl, especially benzyloxycarbonylmethyl, carbamoyl-C1-C6-alkyl, especially carbamoylmethyl, carbamoylethyl, amino-C1-C6-alkyl, especially aminopropyl, aminobutyl, C1-C4-alkylamino-C1-C6-alkyl, especially methylaminopropyl, methylaminobutyl, C1-C4-dialkylamino-C1-C6-alkyl, especially dimethylaminopropyl, dimethylaminobutyl,Guanidino-C1~C6-alkyl, especially guanidinopropyl, C1~C4-alkoxycarbonylamino-C1~C6-alkyl, especially tert-butoxycarbonylaminopropyl, tert-butoxycarbonylaminobutyl, 9-fluorenylmethoxycarbonyl(Fmoc)amino-C1~C6-alkyl, especially 9-fluorenylmethoxycarbonyl(Fmoc)aminopropyl, 9-fluorenylmethoxycarbonyl(Fmoc)aminobutyl, C2~C8-alkenyl, especially vinyl, allyl, butenyl, C3~C7 - Represents cycloalkyl, especially cyclopentyl, cyclohexyl, cycloheptyl, C3-C7-cycloalkyl-C1-C4-alkyl, especially cyclopentylmethyl, cyclohexylmethyl, cycloheptylmethyl, benzyl, substituted benzyl, phenyl, phenyl-C1-C4-alkyl, especially phenylmethyl which may be substituted with a group from the group consisting of halogens, especially fluorine, chlorine, bromine or iodine, hydroxyl, C1-C4-alkoxy, especially methoxy or ethoxy, C1-C4-alkyl, especially methyl.

[0037] In the method according to the present invention, it has been surprisingly found that cyclic depsipeptides (I), particularly emodepsides and closely related structures, can be synthesized in high overall yield using hydrophobic carboxylic acid protecting groups (TAGs). These groups can make the molecules to which they are bound ("tagged" molecules) insoluble in polar solvents such as methanol. Thus, tagged molecules can be precipitated from the reaction mixture, and the tagging groups themselves can also be separated using this technique after deprotection. Furthermore, the hydrophobic tagging groups make tagged molecules soluble in nonpolar solvents such as dichloromethane.

[0038] Deprotection of amine groups by removing the protecting group PG1 can be carried out using standard base-assisted procedures, such as treatment with a piperidine solution in dichloromethane. Similarly, deprotection of carboxylic acid groups by removing the TAG group can be carried out using protocols for removing benzyl groups, such as treatment with a trifluoroacetic acid (TFA) solution in dichloromethane.

[0039] In another embodiment of the method according to the present invention, PG1 is 9-fluorenylmethoxycarbonyl (Fmoc), t-butylcarbamate (Boc), benzylcarbamate (Z), acetamide, trifluoroacetamide, phthalimide, benzyl (Bn), triphenylmethyl (Tr), benzylidene, or p-toluenesulfonamide (Ts),

[0040] TAG [ka] or [ka] (In the formula, m is between 15 and 25, p is between 8 and 18, and q is between 15 and 25.) Preferably, m is 18, 19, 20, 21, or 22, p is 11, 12, or 13, and q is 21, 22, or 23.

[0041] In another embodiment of the method according to the present invention, a depsipeptide according to general formula (IIb) is -In precursor (IVb), the amine group protected by the PG2 group is deprotected in the presence of a base, thereby obtaining the deprotected amine group; -In precursor (IIIb), the carboxylic acid protected by the PG3 group is deprotected in the presence of an acid, thereby obtaining the deprotected carboxylic acid group; - The deprotected amine group and carboxylic acid group are condensed to obtain a depsipeptide (IIb). By doing so, Obtained from precursors according to general formulas (IVb) and (IIIb): [ka] (In the formulas, R1-R12, TAG, PG1, x, and y have the meanings defined above (in particular, x and y may be 1), PG2 is an amine protecting group, and PG3 is a carboxylic acid protecting group). The deprotection and condensation methods may be the same as those outlined in relation to the reaction of compounds (II) to (I) above.

[0042] Preferably, the precursor according to general formula (IVb) is -In precursor (VIb), the amine group protected by the PG4 group is deprotected in the presence of a base, thereby obtaining the deprotected amine group; -The deprotected amine group of precursor (VIb) and the carboxylic acid group of precursor (Vb) are condensed to obtain precursor (IVb). By doing so, Obtained from the precursors of general formula (Vb) and (Vb): [ka] (In the formula, R7-R12, TAG and x have the meanings defined above (in particular, x may be 1), PG2 has the meaning defined above, and PG4 is an amine protecting group). The deprotection and condensation methods may be the same as those outlined in relation to the reaction of compound (IIb) to (I) above.

[0043] The precursor according to general formula (IIIb) is -In precursor (VIIb), the amine group protected by the PG5 group is deprotected in the presence of a base, thereby obtaining the deprotected amine group; - The deprotected amine group of precursor (VIIb) is condensed with the carboxylic acid group of precursor (VIIIb) to obtain precursor (IIIb). By doing so, Obtained from precursors according to general formulas (VIIIb) and (VIIb): [ka] (In the formula, R1 to R6, PG1 and y have the meanings defined above (in particular, y may be 1), PG3 has the meaning defined above, and PG5 is an amine protecting group.) It is also preferable that the deprotection and condensation methods be the same as those outlined in relation to the reaction of compound (IIb) to (I) described above.

[0044] The precursor according to general formula (VIb) is obtained by esterification of the precursor according to general formula (IXb) with TAG-OH: [ka] (In the formula, R10~R12 and TAG have the meanings defined above, and PG4 also has the meanings defined above.) It is also preferable that the carboxylic acid group of (IXb) be protected or tagged using a standard condensation system such as DCC / DMAP.

[0045] The precursor according to general formula (VIIb) is obtained by esterification of the precursor according to general formula (X) with PG3-OH: [ka] (In the formula, R4~R6 and y have the meanings defined above, PG3 also has the meanings defined above, and PG5 also has the meanings defined above.) That is also desirable.

[0046] It is also preferable that PG3 is the TAG defined above.

[0047] In another embodiment of the method according to the present invention, at least one of the reaction steps that yield the TAG-containing molecule is followed by precipitation of the crude reaction product in methanol, thereby purifying the crude reaction product.

[0048] In another embodiment of the method according to the present invention, at least one of the reaction steps in which the TAG-protected carboxylic acid group is deprotected is followed by precipitation of the cleaved TAG-OH in methanol and removal of the precipitate by filtration, thereby purifying the crude reaction product.

[0049] It is also preferable that precursors (IIIb) and (IVb) are the same.

[0050] It is also preferable that R3 and R9 are different from each other, R1 and R7 are the same, R2 and R8 are the same, R4 and R10 are the same, R5 and R11 are the same, and R6 and R12 are the same.

[0051] In another embodiment of the method according to the present invention, the depsipeptide is selected from one of the general formulas (II-1) to (II-14b): [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] (In equations (II-11) to (II-14), -LINK- is [ka] [ka] (In the formula, X1 can be C, N, S, or O, and X2 and X3 can be C or N); [ka] (In the formula, X1 can be C, N, S, or O, and X2, X3, and X4 can be C or N); [ka] (In the formula, X1, X2, X3, and X4 may be C or N) Selected from; R13 is selected from SO2NH(CH3), SO2NH2, OC(O)CH3, CF3, or one of the following lactone structures: [ka] ).

[0052] The present invention further provides a method for synthesizing a cyclic depsipeptide according to general formula (1) from a depsipeptide according to general formula (IIb): [ka] And, -E T (30) A step of preparing a mixture of a solvent compound (IIc) and a base, in which the value is between 30 and 43. - The step of slowly, preferably dropwise, adding a coupling agent solution to a solvent to form a cyclic depsipeptide (I), and Includes, x and y are independently 0, 1, or 2, where x + y ≥ 1 (preferably x and y are 1), and R1, R2, R3, R4, R5, R6, R7, R8, R9, R10, R11, and R12 are each independently hydrogen, linear or branched C1-C8 alkyl, particularly methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, sec-pentyl, hexyl, isohexyl, sec-hexyl, heptyl, isoheptyl, sec-heptyl, tert- Heptyl, octyl, isooctyl, sec-octyl, linear or branched halogenated C1-C8 alkyl, especially sec-butyl fluoride, hydroxy-C1-C6 alkyl, especially hydroxymethyl, 1-hydroxyethyl, C1-C4-alkanoyloxy-C1-C6 alkyl, especially acetoxymethyl, 1-acetoxyethyl, C1-C4-alkoxy-C1-C6 alkyl, especially methoxymethyl, 1-methoxyethyl, aryl-C1-C4-alkyloxy-C1-C6 alkyl, especially benzyloxymethyl, 1-benzyloxyethyl L, mercapto-C1~C6-alkyl, especially mercaptomethyl, C1~C4-alkylthio-C1~C6-alkyl, especially methylthioethyl, C1~C4-alkylsulfinyl-C1~C6-alkyl, especially methylsulfinylethyl, C1~C4-alkylsulfonyl-C1~C6-alkyl, especially methylsulfonylethyl, carboxy-C1~C6-alkyl, especially carboxymethyl, carboxyethyl, C1~C4-alkoxycarbonyl-C1~C6-alkyl, especially methoxycarbonylmethyl, ethoxycarbonylethyl, C1~C 4-arylalkoxycarbonyl-C1~C6-alkyl, especially benzyloxycarbonylmethyl, carbamoyl-C1~C6-alkyl, especially carbamoylmethyl, carbamoylethyl, amino-C1~C6-alkyl, especially aminopropyl, aminobutyl, C1~C4-alkylamino-C1~C6-alkyl, especially methylaminopropyl, methylaminobutyl, C1~C4-dialkylamino-C1~C6-alkyl, especially dimethylaminopropyl, dimethylaminobutyl, guanidino-C1~C6-alkyl, especially guanidinopropyl,C1-C4-alkoxycarbonylamino-C1-C6-alkyl, particularly tert-butoxycarbonylaminopropyl, tert-butoxycarbonylaminobutyl, 9-fluorenylmethoxycarbonyl(Fmoc)amino-C1-C6-alkyl, particularly 9-fluorenylmethoxycarbonyl(Fmoc)aminopropyl, 9-fluorenylmethoxycarbonyl(Fmoc)aminobutyl, C2-C8-alkenyl, particularly vinyl, allyl, butenyl, C3-C7-cycloalkyl, particularly cyclopentyl, cyclohexyl, cycloheptyl, C3-C7-cycloalkyl-C1-C4-alkyl, particularly cyclopentylmethyl, cyclohexylmethyl, cycloheptylmethyl, benzyl, substituted benzyl, phenyl, phenyl-C1-C4-alkyl, particularly phenylmethyl which may be substituted with a group from the group consisting of halogens, particularly fluorine, chlorine, bromine or iodine. Provide a method.

[0053] The term "slowly" means and / or includes, in particular, that the coupling agent solution is added at a rate of 2(mol)-% or less per minute, preferably 1(mol)-% or less per minute, and more preferably 0.5(mol)-% or less per minute.

[0054] Surprisingly, the method according to the present invention has been found to enable the synthesis of cyclic depsipeptides (I), particularly emodepsides and closely related structures, in high overall yield. While not bound by any theory, the inventors believe this is due to the low solubility of cyclic depsipeptides (I) in solvents, in contrast to the ring-opened form (IIb).

[0055] "E T (30) The term “value” is used by Reichardt, Angew. Chem. 1979, 119-131, which lists both methods for determining such values ​​and measurements for many standard solvents. T (30) refers to this.

[0056] According to one embodiment of the present invention, the solvent ET (30) The value is between 34 and 39.

[0057] In the meaning of this invention, the term "solvent" also includes mixtures of solvents.

[0058] According to one embodiment of the present invention, the solvent contains ethyl acetate, and according to one embodiment of the present invention, the solvent is ethyl acetate.

[0059] A suitable coupling agent in an embodiment of the present invention is shown above. According to one embodiment of the present invention, the coupling agent comprises T3P, and according to one embodiment of the present invention, the coupling agent is T3P.

[0060] According to one embodiment, the ratio (moles:moles) of the coupling agent to compound (IIc) before the reaction is 1:1 or more and 5:1 or less, preferably 2:1 or more and 3:1 or less.

[0061] According to one embodiment, the ratio (moles:moles) of the base to compound (IIb) before the reaction is 2:1 or more and 10:1 or less, preferably 4:1 or more and 6:1 or less.

[0062] Suitable bases insofar as they are embodiments of the present invention include N,N-diisopropylethylamine (DIEA), triethylamine, dimethylaminopyridine (DMAP), N-methylmorpholine, pyridine, 1,8-diazabicyclo[5.4.0]undeca-7-ene (DBU), 1,5-diazabicyclo[4.3.0]nona-5-ene (DBN), or mixtures thereof. N,N-diisopropylethylamine is most preferred.

[0063] According to one embodiment, the temperature during the addition of the coupling agent is 25°C or lower.

[0064] In one embodiment of the method according to the present invention, x is 1, y is 1, R1, R4, R7 and R10 are methyl, R6 and R12 are methyl, R5 and R11 are independently linear or branched C1-C4 alkyl or linear or branched halogenated C1-C4 alkyl, and R3 and R9 are independently benzyl or substituted benzyl. According to one preferred embodiment of the invention, R3 and / or R9 is p-morpholino-substituted benzyl.

[0065] In one embodiment of the method according to the present invention, R3 and R9 are the same, R1 and R7 are the same, R2 and R8 are the same, R4 and R10 are the same, R5 and R11 are the same, and R6 and R12 are the same.

[0066] According to a preferred embodiment of the present invention, compound IIb is synthesized from compound II or IIa by deprotecting protecting groups A and B or X and Y, respectively. Compound IIb can be isolated or used in situ to synthesize depsipeptide (I).

[0067] The present invention also relates to linear or cyclic depsipeptides selected from one of the general formulas (II-1) to (II-14b) or (I-1) to (I-9) shown below, or to pharmaceutically or veterinarily acceptable salts thereof: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] (In equations (II-11) to (II-14) and (I-1) to (I-9), respectively, A, B, X, and Y are defined as above. -LINK- is [ka] Selected from, R13 is selected from SO2NH(CH3), SO2NH2, OC(O)CH3, CF3, or one of the following lactone structures: [ka] ).

[0068] The terms “veterinary acceptable salt” and “pharmaceutical acceptable salt” are used throughout this specification to describe any salt of a compound that is acceptable for administration for medicinal or veterinary use and that provides an active compound at the time of administration.

[0069] Veterinary-acceptable salts include those derived from veterinary-acceptable inorganic or organic bases and acids. Suitable salts include those containing alkali metals such as lithium, sodium, or potassium, and alkaline earth metals such as calcium, magnesium, and barium. However, salts containing transition metals, including manganese, copper, zinc, and iron, are also suitable. Furthermore, salts containing ammonium cations and substituted ammonium cations, in which one or more hydrogen atoms are substituted by alkyl or aryl groups, are also included in this invention. However, salts derived from inorganic acids, including hydrohalic acids (HCl, HBr, HF, HI), sulfuric acid, nitric acid, and phosphoric acid, are particularly suitable. Suitable inorganic salts also include, but are not limited to, bicarbonates and carbonates. In some embodiments, examples of veterinary and agriculturally acceptable salts are organic acid addition salts formed from organic acids, including, but not limited to, maleates, dimaleates, fumarates, tosylates, methanesulfons, acetates, citrates, malons, tartrates, succinates, benzoates, ascorbicates, α-ketoglutarates, and α-glycerophosphates. Of course, other acceptable organic acids may also be used.

[0070] Alkali metal (e.g., sodium, potassium, or lithium) or alkaline earth metal (e.g., calcium) salts of compounds can also be produced by reacting a sufficiently acidic residue on the compound with an alkali metal or alkaline earth metal hydroxide.

[0071] Veterinary-acceptable salts can be obtained using standard procedures well known in the art, for example, by reacting a sufficiently basic compound, such as an amine, with a suitable acidic functional group present in the compound, or by reacting a suitable acid with a suitable basic functional group on the compound of the present invention. [Brief explanation of the drawing]

[0072] [Figure 1] This document describes a synthesis scheme for emodepsid using benzyl(R)-2-hydroxy-3-(4-morpholinophenyl)propanoate (EMD-8), a compound synthesized from p-fluorobenzaldehyde. [Figure 2-1] The synthesis scheme for emodepside is described below. [Figure 2-2] See Figure 2-1. [Figure 2-3] See Figure 2-1. [Figure 2-4] See Figure 2-1. [Figure 2-5] See Figure 2-1. [Figure 2-6] See Figure 2-1. [Figure 3-1] The reaction scheme for PF1022A is described below. [Figure 3-2] See Figure 3-1. [Figure 3-3] See Figure 3-1. [Figure 4] The NMR spectrum of PF1022A is described below. [Figure 5-1] The reaction scheme for PF1022A is described below. [Figure 5-2] See Figure 5-1. [Figure 5-3] See Figure 5-1. [Figure 6-1] The reaction scheme for emodepside is described below. [Figure 6-2] See Figure 6-1. [Figure 6-3] See Figure 6-1. [Figure 6-4] See Figure 6-1. [Figure 7] The NMR spectrum of the emodepside is described below. [Figure 8] The LC-UV spectrum of emodepside is described. [Figure 9-1] The reaction scheme for emodepside is described below. [Figure 9-2] See Figure 9-1. [Figure 9-3] See Figure 9-1. [Modes for carrying out the invention]

[0073] Examples The present invention will be further described with reference to the following embodiments, but will not be limited thereto.

[0074] 1. General method Starting structural units were prepared from commercially available reagents. Unless otherwise noted, reagents and solvents were purchased at the highest commercial quality and used without further purification. Methanol, anhydrous toluene, and CH2Cl2 were purchased from Kanto Chemical Co., Ltd. All reactions were monitored by thin-layer chromatography (TLC) using Merck silica gel 60 F254 pre-coated plates (0.25 mm). Flash chromatography was performed using Kanto Chemical silica gel (Kanto Chemical, Silica Gel 60N, spherical neutral, 0.040-0.050 mm, catalog number 37563-84). 1 H and 13 The 1C NMR spectrum was obtained using JEOL JNM-ECA-500. 1 For H-NMR, 500MHz, 13 The 1C-NMR spectrum was recorded at 125MHz. The chemical shift was CDCl3( 1 H;δ=7.26ppm, 13 C;δ=77.0ppm), CD3OD( 1 H;δ=3.31ppm, 13The spectrum is expressed as downfield ppm from the internal solvent peak for C (δ=49.0 ppm), and the J value is given in Hertz. The following abbreviations were used to describe multiplicity: s=singlet, d=doublet, t=triplet, q=quadruplet, dd=doubledoublet, ddd=doubledoubledoublet, dt=doubletriplet, dq=doublequadruplet, m=multitlet, br=broad. All infrared spectra were measured with a Horiba FT-210 spectrometer. High-resolution and low-resolution mass spectra were measured with a JEOL JMS-AX505 HA, JEOL JMS-700 MStation, and JEOL JMS-T100LP. Optical rotation was measured using a JASCO P-1010 polarimeter. Melting point was measured with a YANACO MP-500P or OptiMelt (Stanford Research Systems) instrument.

[0075] Common methods for Fmoc deprotection The Fmoc protecting substrate was dissolved in 5% piperidine / CH2Cl2 (typically 0.05 M relative to the substrate) at room temperature, and the solution was stirred for 3 hours. The reaction mixture was then cooled to -5°C, and MeOH was added (in a 5-fold excess of the reaction solution). The resulting heterogeneous solution was stirred for a further 30 minutes at -5°C, the colorless precipitate was filtered, and washed with additional MeOH to obtain the corresponding amine as a colorless powder.

[0076] Common methods for cleaving TAGa functional groups The TAGa-tagged substrate was dissolved in 50% TFA / CH2Cl2 (0.05 M relative to the substrate) at room temperature, and the solution was stirred for approximately 1 hour. The reaction mixture was then concentrated with toluene (x3) to remove the TFA. Next, CH2Cl2 was added to the flask at -5°C, followed by MeOH (5-fold excess CH2Cl2). The resulting heterogeneous solution was stirred for a further 30 minutes at -5°C, and the colorless precipitate was filtered off and washed with additional MeOH. The combined filtrate was concentrated under vacuum. 4 M HCl / dioxane (0.05 M relative to the product) was added to the resulting product, and the mixture was concentrated with toluene (x3) to obtain the corresponding carboxylic acid as a generally brown oil. The crude product was used in the next reaction without further purification.

[0077] The product was concentrated with toluene (x3) to obtain the corresponding carboxylic acid as a generally brown oil. The crude product was used in the next reaction without further purification.

[0078] 2. Synthesis of Emodepsides The synthesis of emodepside using benzyl(R)-2-hydroxy-3-(4-morpholinophenyl)propanoate (EMD-8), a compound synthesized from p-fluorobenzaldehyde, is described below according to the scheme in Figure 1. Using this compound, emodepside is synthesized according to the scheme in Figure 2. [ka]

[0079] 4-Molfolinobenzaldehyde (EMD-22): In a 100 L reactor, 4-fluorobenzaldehyde (EMD-21, 3.6 kg, 29.0 mol, 1.0 equivalent) in 1-methyl-2-pyrrolidine (36 L, 10.0 V), morpholine (7.6 kg, 87 mol, 3.0 equivalent), and K2CO3 (10.0 kg, 72.5 mol, 2.5 equivalent) were added. The resulting mixture was stirred at 125-130°C for at least 6 hours. The reaction was monitored by TLC until EMD-21 was gone. The reaction mixture was diluted with ethyl acetate (18 L, 5 V) and H2O (72 L, 20 V) and separated. The aqueous phase was extracted with ethyl acetate (18 L x 2), the organic phase was combined, and washed with H2O (36.0 L x 3). The organic extract was concentrated under vacuum at a temperature below 45°C until no more distillate spilled. The residue was eluted with heptane / ethyl acetate (5:1, v / v, 7.2 L) and concentrated under vacuum at a temperature below 45°C. Then, heptane / ethyl acetate (5:1, v / v, 21.6 L) was added to the residue at 20-25°C. The solution was stirred at 20-25°C for 16 hours. The mixture was filtered, and the filter cake was washed with heptane (7.3 L). The solid was dried under vacuum at 40-45°C. This yielded 4.67 kg (84.8%) of 4-morpholinobenzaldehyde (EMD-22) as a yellow solid. MS(ES,m / z):192(M+H); 1H NMR (DMSO-d6,300 MHz)9.74(s,1H),7.74(d,J=8.1 Hz,2H),7.07(d,J=8.4 Hz,2H),3.75-3.74(m,4H),3.35-3.33(m,4H). [ka]

[0080] (Z)-2-methyl-4-(4-morpholinobenzylidene)oxazole-5(4H)-one (EMD-22B): In a 100 L reactor, N-acetylglycine (1.22 kg, 10.46 mol, 1.0 equivalent) in tetrahydrofuran (20 L, 10 V), acetic anhydride (3.2 kg, 31.38 mol, 3.0 equivalent), and zinc(II) chloride (1.48 kg, 10.46 mol, 1.0 equivalent) were added. The resulting mixture was stirred at 70°C for 1 hour, and EMD-22 (2.0 kg, 10.46 mol, 1.0 equivalent) was added. The mixture was then stirred at 70°C for a further 16 hours and monitored by LCMS. After cooling to 20-25°C, H2O (40 L) was added. The mixture was then stirred at 0-5°C for 3 hours and filtered. The filtered cake was washed with H2O (10 L) and dried under vacuum at 40-45°C. This yielded 2.29 kg (80.4%) of (Z)-2-methyl-4-(4-morpholinobenzylidene)oxazole-5(4H)-one (EMD-22B) as a brown solid. MS(ES, m / z): 273(M+H); 1 H NMR (DMSO-d6,300 MHz)7.84(s,1H),7.47(d,J=9.0 Hz,2H),7.02(d,J=9.1 Hz,2H),3.74-3.71(m,4H),3.32-3.27(m,4H). [ka]

[0081] (E)-2-hydroxy-3-(4-morpholinophenyl)acrylic acid (EMD-23): In a 50 L reactor, EMD-22B (2.2 kg, 8.08 mol, 1.0 equivalent) in 1,4-dioxane (8.8 L, 4.0 V) and HCl (8.8 L, 4.0 V) were added at 20-25°C. The resulting mixture was stirred at 80°C for 3 hours and monitored by LC-MS. After cooling to 0-10°C, the mixture was stirred at 0-10°C for 16 hours and filtered. The filtered cake was dried under vacuum at 40-45°C. The crude product was eluted with H2O (4.4 L) and stirred at 0-10°C for 2 hours. The mixture was filtered and the filtered cake was dried under vacuum at 40-45°C. This yielded 1.17 kg (56.0%) of (E)-2-hydroxy-3-(4-morpholinophenyl)acrylic acid (EMD-23) as a Slater solid. MS(ES,m / z):250(M+H); 1 H NMR (DMSO-d6,300 MHz)7.66(d,J=8.5 Hz,2H),6.97(d,J=8.6 Hz,2H),6.35(s,1H),4.05(s,1H,-OH),3.77-3.74(m,4H),3.19-3.16(m,4H). [ka]

[0082] (R)-2-hydroxy-3-(4-morpholinophenyl)propanoic acid (EMD-24): In a 2 L round-bottom flask purged and maintained under an inert nitrogen atmosphere, EMD-23 (66.0 g, 0.26 mol, 1.0 equivalent) in a solution of N,N-dimethylformamide (660 mL, 10.0 V), Et3N (107.2 g, 1.06 mol, 4.0 equivalents), and RuCl(R,R)-TsDPEN (1.69 g, 0.0026 mol, 0.01 equivalent) were added at 20-25°C. Formic acid (36.59 g, 0.79 mol, 3.0 equivalents) was added dropwise to the above mixture under an N2 atmosphere at 20-25°C for 2 hours. The mixture was then stirred at 20-25°C and monitored by LC-MS. The reaction product was identified as M1 and used in the next step without further purification. [ka]

[0083] Benzyl (R)-2-hydroxy-3-(4-morpholinophenyl)propanoate (EMD-8): To the mixture (M1), K2CO3 (109.0 g, 0.78 mol, 3.0 equivalents) was added at 20 - 25 °C for 1 hour, and benzyl bromide (54.0 g, 0.32 mol, 1.2 equivalents) was added dropwise. Then, the mixture was stirred at 55 - 60 °C for an additional 16 hours. The reaction mixture was diluted with ethyl acetate (330 mL, 5V) and H2O (1.2 L, 20V) and separated. The aqueous phase was extracted with ethyl acetate (330 mL x 2), and the organic phases were combined and washed with H2O (660 mL x 3). The organic extract was concentrated under vacuum at below 45 °C until no distillate dripped out. The residue was eluted with heptane / ethyl acetate (3:1, v / v, 132 mL) and concentrated under vacuum at below 45 °C. Then, heptane / ethyl acetate (3:1, v / v, 264 mL) was added to the above residue at 20 - 25 °C. The mixture was filtered, and the filter cake was washed with heptane (132 mL). The solid was dried under vacuum at 40 - 45 °C. Thus, 52 g (58%) of benzyl (R)-2-hydroxy-3-(4-morpholinophenyl)propanoate (EMD-8) was obtained as a yellow solid. MS (ES, m / z): 342 (M+H); 1 H NMR (DMSO-d6, 300 MHz) 7.40 - 7.27 (m, 5H), 7.05 (d, J = 8.1 Hz, 2H), 6.82 (d, J = 8.1 Hz, 2H), 5.17 (s, 2H), 4.24 (t, J = 7.5 Hz, 1H), 3.74 - 3.71 (m, 4H), 3.06 - 3.03 (m, 4H), 2.90 - 2.73 (m, 2H).

Chemical Structure

[0084] (R)-1-(benzyloxy)-3-(4-morpholinophenyl)-1-oxopropan-2-yl-N-(tert-butoxycarbonyl)-N-methyl-L-leucinate (EMD-9B): In a 50 L reactor purged and maintained under an inert nitrogen atmosphere, EMD-8 (1.96 kg, 5.75 mol, 1.0 equivalent) in dichloromethane (14.1 L, 10 V), N-(tert-butoxycarbonyl)-N-methyl-L-leucine (1.41 kg, 5.75 mol, 1.0 equivalent), DMAP (0.77 kg, 6.32 mol, 1.1 equivalent), and EDCI (1.21 kg, 6.32 mol, 1.1 equivalent) were added at 20-25°C. The mixture was stirred at this temperature for at least 3 hours and monitored by LCMS. The mixture was concentrated at a temperature below 40°C until no more distillate spilled. The residue was dissolved in MTBE (14.1 L) and HCl (aqueous solution, 1N, 14.1 L) and filtered. The organic matter was washed with HCl (aqueous solution, 1N, 14.1 L) and NaHCO3 (saturated, 14.1 L x 2), and concentrated at a temperature below 40°C until no more distillate spilled. The residue was then dissolved in heptane / MTBE (28.2 L, 8:1, v / v) and concentrated at a temperature below 40°C until no more distillate spilled. Heptane / MTBE (15.5 L, 8:1, v / v) was added to the above residue at 20-25°C, seed crystals (0.2%, w / w) were added, and the mixture was stirred overnight at 20-25°C. The mixture was filtered; the filtered cake was washed with heptane (7.0 L). The solid was dried under vacuum at 40-45°C. This yielded 2.56 kg (78.3%) of (R)-1-(benzyloxy)-3-(4-morpholinophenyl)-1-oxopropan-2-yl-N-(tert-butoxycarbonyl)-N-methyl-L-leucinate (EMD-9B) as a pale yellow solid. MS(ES,m / z):569(M+H); 1H NMR(DMSO-d6,300 MHz)7.38-7.26(m,5H),7.10(d,J=8.2 Hz,2H),6.96(d,J=8.1 Hz,2H),5.25-5.22(m,1H),5.12-5.10(m,2H),4.07-3.99(m,1 H),3.79-3.76(m,,4H),3.19-2.97(m,6H),2.57(s,3H),1.42-1.34(m,11H),1.24-1.15(m,1H),0.88-0.82(m,6H). [ka]

[0085] (R)-2-((N-(tert-butoxycarbonyl)-N-methyl-L-leucyl)oxy)-3-(4-morpholinophenyl)propanoic acid (EMD-10B): In a 50 L reactor purged and maintained under an inert nitrogen atmosphere, a solution of EMD-9B (400 g, 0.7 mol, 1.0 equivalent) in EtOH (4.0 L, 10.0 V) was added at 20-25°C. The reactor was evacuated, nitrogen was passed through three times, and Pd / C (28.0 g, 7% w / w) was added. The reactor was then evacuated again, nitrogen was passed through three more times, and hydrogen bubbling was maintained below the surface of the reaction mixture. The mixture was stirred at 20-25°C for at least 4 hours and monitored by HPLC. After the reaction was complete, hydrogen bubbling was stopped. The mixture was filtered through celite® (2.0 kg), the filtration cake was rinsed with EA (0.8 L), and the filtrate was concentrated at below 40°C until no more distillate spilled. This yielded 317.7 g (94.4%) of (R)-2-((N-(tert-butoxycarbonyl)-N-methyl-L-leucyl)oxy)-3-(4-morpholinophenyl)propanoic acid (EMD-10B) as a dark yellow oil; MS(ES, m / z): 479(M+H); 11H NMR (DMSO-d6, 300 MHz) δ 7.10 (d, J = 8.4 Hz, 2H), 6.85 (d, J = 8.7 Hz, 2H), 5.03 - 5.02 (m, 1H), 4.81 - 4.53 (m, 1H), 3.73 (t, J = 7.2 Hz, 4H), 3.05 (t, J = 7.2 Hz, 4H), 2.96 - 2.90 (m, 1H), 2.63 (s, 3H), 1.42 - 1.16 (m, 12H), 0.89 - 0.84 (m, 6H).

Chem.

[0086] (R)-1-(benzyloxy)-1-oxopropan-2-ylmethyl-L-leucinate (EMD-12B): In a 10 L reactor purged and maintained under an inert nitrogen atmosphere, a solution of N-(tert-butoxycarbonyl)-N-methyl-L-leucine (240.3 g, 0.98 mol, 1.0 equivalent) in THF (3.9 L, 16 V), benzyl(S)-2-hydroxypropanoate (176.5 g, 0.98 mol, 1.0 equivalent), and triphenylphosphine (385.1 g, 1.47 mol, 1.5 equivalent) were added at 20-25°C. After cooling to below 10°C, diisopropyl azodiformate (297 g, 1.47 mol, 1.5 equivalent) was added dropwise with stirring at below 10°C. Next, the mixture was heated to 20-25°C and stirred for at least 2 hours. The reaction was monitored by HPLC until the benzyl(S)-2-hydroxypropanoate content was 0.5% or less. The resulting mixture was diluted with ethyl acetate (3.9 L) and washed with saturated NaHCO3 (3.9 L x 2) and saline solution (3.9 L x 2). The organic phase was concentrated at a temperature below 40°C until no more distillate spilled. The residue was replaced with tert-butyl methyl ether (240 mL x 2), concentrated at a temperature below 40°C until no more distillate spilled, and slurried with tert-butyl methyl ether (960 mL) for at least 3 hours. The mixture was filtered, and the filtered cake was washed with tert-butyl methyl ether (240 mL). The filtrate was concentrated at a temperature below 40°C until no more distillate spilled. This yielded the crude product (R)-1-(benzyloxy)-1-oxopropan-2-yl N-(tert-butoxycarbonyl)-N-methyl-L-leucinate (EMD-1B) as a yellow oil, which was used in the next step without further purification.

[0087] A 10 L reactor, purged and maintained under an inert nitrogen atmosphere, was given a 1.2 L (5.0 equivalent) solution of EMD-1B in HCl / EA at a temperature below 25°C. The mixture was stirred at 20–25°C for at least 1 hour and monitored by HPLC until the EMD-1B content was less than 0.5%. The solution was concentrated at a temperature below 40°C until no more distillate spilled. The residue was replaced with tert-butyl methyl ether (240 mL x 2), concentrated at a temperature below 40°C until no more distillate spilled, and dissolved in tert-butyl methyl ether (1.92 L). Seed crystals (0.1%, w / w) were then added and stirred at 20–25°C for at least 5 hours. The mixture was filtered, and the filter cake was washed with tert-butyl methyl ether (0.24 L). The solid was dried under vacuum at 40±5°C to obtain 270 g of crude product as a white solid. The solid was dissolved in ethyl acetate (810 mL) by heating to 40±5°C, and tert-butyl methyl ether (4.05 L) was added. The mixture was then cooled to 20-25°C, and seed crystals (0.1% w / w) were added. The mixture was stirred at 20-25°C for at least 5 hours, filtered, and the filtered cake was washed with tert-butyl methyl ether (0.24 L). The solid was dried under vacuum at 40±5°C. This yielded 226.7 g (67.3%, 2 steps) of (R)-1-(benzyloxy)-1-oxopropan-2-ylmethyl-L-leucinate (EMD-12B) as a white solid. MS(ES,m / z):308(M+H); 1 H NMR(DMSO-d6,300 MHz)7.41-7.35(m,5 H),5.28(q,J=7.1 Hz,1H),5.20(s,2H),2.51(s,3H),1.77-1.71(m,3H),1.50-1.48(m,3H),0.90-0.87(m,6H). [ka]

[0088] (R)-1-(benzyloxy)-1-oxopropan-2-yl N-((R)-2-((N-(tert-butoxycarbonyl)-N-methyl-L-leucyl)oxy)-3-(4-morpholinophenyl)propanoyl)-N-methyl-L-leucinate (EMD-13B): In a 10 L reactor purged and maintained under an inert nitrogen atmosphere, EMD-10B (275.0 g, 0.57 mol, 1.0 equivalent) in ethyl acetate (2.2 L, 8.0 V), EMD-12B (197.7 g, 0.57 mol, 1.0 equivalent), and N,N-diisopropylethylamine (372.2 g, 2.88 mol, 5.0 equivalent) were added at 20-25°C. After cooling to 10–20°C, propylphosphonic anhydride (916.4 g, 1.44 mol, 2.5 equivalents) was added dropwise with stirring at below 25°C. The mixture was then stirred at 20–25°C for at least 2.5 hours and monitored by HPLC until EMD-12B was less than 1.0%. The solution was diluted with heptane (2.75 L) at 0–10°C and slowly quenched with HCl (aqueous solution, 1.0 N, 2.75 L). The organic phase was washed with HCl (aqueous solution, 1.0 N, 2.75 L) and NaHCO3 (saturated, 2.75 L x 2) and concentrated at below 40°C until no more distillate spilled. As a result, 429.8 g (97.4%) of (R)-1-(benzyloxy)-1-oxopropan-2-yl N-((R)-2-((N-(tert-butoxycarbonyl)-N-methyl-L-leucyl)oxy)-3-(4-morpholinophenyl)propanoyl)-N-methyl-L-leucinate (EMD-13B) was obtained as a yellow concentrated oil. MS(ES,m / z):768(M+H); [ka] (R)-1-(benzyloxy)-1-oxopropan-2-yl-N-methyl-N-((R)-2-((methyl-L-leucyl)oxy)-3-(4-morpholinophenyl)propanoyl)-L-leucinate (EMD-14B): A solution of EMD-13B (245 g, 0.32 mol, 1.0 equivalent) in HCl / EA (735, 3.0 V) was added to a 5 L reactor purged and maintained under an inert nitrogen atmosphere at below 25°C. The mixture was stirred at 20-25°C for at least 1 hour and monitored by HPLC until the EMD-13B content was 0.5% or less. The resulting solution was concentrated at below 40°C until no more distillate spilled, and then replaced with ethyl acetate (245 mL x 2) at below 40°C. The residue was dissolved in ethyl acetate (735 mL), and N,N-diisopropylethylamine (245 g) was added at 20-25°C. The mixture was washed with NaHCO3 (saturated, 735 mL x 2). The organic phase was concentrated at below 40°C until no more distillate spilled. This yielded 186.9 g (89.9%) of (R)-1-(benzyloxy)-1-oxopropan-2-yl-N-methyl-N-((R)-2-((methyl-L-leucyl)oxy)-3-(4-morpholinophenyl)propanoyl)-L-leucinate (EMD-14B) as a yellow oil. MS(ES,m / z):668(M+H); 1 H NMR(DMSO-d6,300 MHz)7.41-7.32(m,5 H),7.17(d,J=8.5 Hz,2H),6.86(d,J=8.6 Hz,2H),5.50-5.47(m,1H),5.20-5.04(m,4H),4.23-3.99(m,1H),3.74-3.72(m,4H),3.09- 2.76(m,10H),2.22-1.99(m,3H),1.63-1.35(m,5H),1.29-1.16(m,4H),0.97-0.70(m,12H). [ka]

[0089] (6S,9R,12S,15R)-6,12-diisobutyl-2,2,5,11,15-pentamethyl-9-(4-morpholinobenzyl)-4,7,10,13-tetraoxo-3,8,14-trioxa-5,11-diazahexadecane-16-acid (EMD-15B): In a 10 L reactor purged and maintained under an inert nitrogen atmosphere, a solution of EMD-13B (274.2 g, 0.36 mol, 1.0 equivalent) in EtOH (2.8 L, 10.0 V) was added at 20-25°C. The reactor was then evacuated, nitrogen was passed through three times, and Pd / C (19.2 g, 7% w / w) was added. The reactor was then evacuated again, nitrogen was passed through three more times, and hydrogen bubbling was maintained below the surface of the reaction mixture. The mixture was stirred at 20-25°C for at least 4 hours and monitored by HPLC. After the reaction was complete, hydrogen bubbling was stopped. The mixture was filtered through celite® (2.0 kg), the filtrate was rinsed with ethyl acetate (0.56 L), and the filtrate was concentrated at below 40°C until no more distillate spilled. This yielded 237.4 g (98.0%) of (6S,9R,12S,15R)-6,12-diisobutyl-2,2,5,11,15-pentamethyl-9-(4-morpholinobenzyl)-4,7,10,13-tetraoxo-3,8,14-trioxa-5,11-diazahexadecane-16-acid (EMD-15B) as yellow oil; MS(ES,m / z):678(M+H); 1 H NMR(DMSO-d6,300 MHz)7.15(d,J=8.4 Hz,2H),6.85(d,J=8.3 Hz,2H),5.52-5.40(m,1H),5.09-5.02(m,1H),4.91-4.53(m,2 H),3.74-3.72(m,4H),3.15-3.04(m,5H),2.94-2.86(m,4H),2.65-2.64(m,3H),1.43-1.28(m,16H),0.93-0.77(m,12H). [ka]

[0090] (R)-1-(benzyloxy)-1-oxopropan-2-yl N-methyl-N-((6S,9R,12S,15R,18S,21R)-6,12,18-triisobutyl-2,2,5,11,15,17-hexamethyl-9,21-bis(4-morpholinobenzyl)-4,7,10,13,16,19-hexaoxo-3,8,14,20-tetraoxa-5,11,17-triazadocosan-22-oil)-L-leucinate (EMD-16B): In a 10 L reactor purged and maintained under an inert nitrogen atmosphere, EMD-15B (147.6 g, 0.22 mol, 1.0 equivalent) in ethyl acetate (2.2 L, 8.0 V) and EMD-14B (145.5 N,N-diisopropylethylamine (139.6 g, 1.08 mol, 5.0 equivalents) was added at 20-25°C. After cooling to 10-20°C, propylphosphonic anhydride (346.4 g, 0.54 mol, 2.5 equivalents) was added dropwise with stirring at below 25°C. The mixture was then stirred at 20-25°C for at least 2.5 hours and monitored by HPLC until EMD-12B was less than 0.5%. The solution was diluted with heptane (2.2 L) at 0-10°C and slowly quenched with HCl (aqueous solution, 1.0 N, 2.2 L). The organic phase was washed with HCl (aqueous solution, 1.0 N, 2.2 L) and NaHCO3 (saturated, 2.2 L x 2), concentrated at below 40°C until no more distillate spilled, and replaced with heptane / MTBE (0.44 L, 1.5:1, v / v) at below 40°C. The residue was dissolved in heptane / MTBE (1.65 L, 1.5:1, v / v), and seed crystals (0.1%, w / w) were added at 20-25°C. The mixture was stirred at 20-25°C for at least 16 hours and filtered. The filtered cake was washed with heptane (0.66 L) and dried under vacuum at 40-45°C.As a result, 242.4 g (83.4%) of (R)-1-(benzyloxy)-1-oxopropan-2-yl N-methyl-N-((6S,9R,12S,15R,18S,21R)-6,12,18-triisobutyl-2,2,5,11,15,17-hexamethyl-9,21-bis(4-morpholinobenzyl)-4,7,10,13,16,19-hexaoxo-3,8,14,20-tetraoxa-5,11,17-triazadocosan-22-oil)-L-leucinate (EMD-16B) was obtained as a white solid: MS (ES, m / z): 1328 (M+H). [ka] (R)-1-(benzyloxy)-1-oxopropan-2-ylN-((2R,5S,8R,11S,14R,17S)-5,11-diisobutyl-6,8,12,19 -Tetramethyl-17-(methylamino)-2,14-bis(4-morpholinobenzyl)-4,7,10,13,16-pentaoxo-3,9,15-trioxa-6,12-diazycosanoyl)-N-methyl-L-leucinate (EMD-20B): A solution of EMD-16B (376.4 g, 0.28 mol, 1.0 equivalent) in HCl / EA (1128 mL, 3.0 V) was added to a 5 L reactor purged and maintained under an inert nitrogen atmosphere at below 25°C. The mixture was stirred at 20-25°C for at least 1 hour and monitored by HPLC until the EMD-16B content was 0.5% or less. The resulting solution was concentrated at below 40°C until no more distillate spilled, and then replaced with ethyl acetate (376.4 mL × 2) at below 40°C. The residue was dissolved in ethyl acetate (1128 mL), and N,N-diisopropylethylamine (376.4 g) was added at 20-25°C. The mixture was washed with NaHCO3 (saturated, 1128 mL x 2). The organic phase was concentrated at a temperature below 40°C until no more distillate spilled. As a result, (R)-1-(benzyloxy)-1-oxopropan-2-yl N-((2R,5S,8R,11S,14R,17S)-5,11-diisobutyl-6,8,12,19-tetramethyl-17-(methylamino)-2,14-bis(4-morpholinobenzyl)-4,7,10,13,16-pentaoxo-3,9,15-trioxa-6,12-diazycosanoyl)-N-methyl-L-leucinate (EMD-20B) 313.56 g (90.1%) was obtained as yellow oil; MS(ES,m / z):1228(M+H); [ka] (3S,6R,9S,12R,15S,18R,21S,24R)-3,9,15,21-tetraisobutyl-8,12,14,20,24-pentamethyl-6,18-bis(4-morpholinobenzyl)-4,7,10,13,16,19,22-heptaoxo-5,11,17,23-tetraoxa-2,8,14,20-tetraazapentacosan-25-acid (EMD-18B): Add a solution of EMD-20B (313.56 g, 0.26 mol, 1.0 equivalent) in EtOH (3.2 L, 10.0 V) at 20-25°C to a 10 L reactor purged and maintained under an inert nitrogen atmosphere. Empty the reactor, pass nitrogen through three times, and adjust Pd / C (21.95 g, 7% w / w) was added. The reactor was then evacuated, nitrogen was passed through it three more times, and hydrogen bubbling was maintained below the surface of the reaction mixture. The mixture was stirred at 20-25°C for at least 4 hours and monitored by HPLC until EMD-20B was less than 1.0%. After the reaction was complete, hydrogen bubbling was stopped. The mixture was filtered through celite® (2.0 kg), the filtrate was rinsed with ethyl acetate (0.64 L x 3), and the filtrate was concentrated at below 40°C until no more distillate spilled. The residue was slurried with ethyl acetate (0.7 L) for at least 3 hours and filtered; the filtrate was rinsed with ethyl acetate (0.32 L x 2). The solid was dried under vacuum at 40-45°C. As a result, 234.2 g (80.6%) of (3S,6R,9S,12R,15S,18R,21S,24R)-3,9,15,21-tetraisobutyl-8,12,14,20,24-pentamethyl-6,18-bis(4-morpholinobenzyl)-4,7,10,13,16,19,22-heptaoxo-5,11,17,23-tetraoxa-2,8,14,20-tetraazapentacosan-25-acid (EMD-18B) was obtained as a grayish-white solid; MS(ES,m / z):1138(M+H); 11H NMR (DMSO-d6, 300 MHz) δ 7.34 - 7.16 (m, 8H), 5.68 - 5.28 (m, 3H), 5.11 - 5.04 (m, 3H), 4.93 - 4.86 (m, 1H), 4.03 - 3.99 (m, 1H), 3.85 - 3.82 (m, 8H), 3.23 - 3.22 (m, 8H), 3.23 - 2.76 (m, 13H), 2.49 (s, 2H), 2.07 (s, 2H), 1.66 - 1.45 (m, 8H), 1.43 - 1.16 (m, 10H), 0.99 - 0.64 (m, 24H). [Chemical formula]

[0091] (3S,6R,9S,12R,15S,18R,21S,24R)-3,9,15,21-tetraisobutyl-4,6,10,16,18,22-hexamethyl-12,24-bis(4-morpholinobenzyl)-1,7,13,19-tetraoxa-4,10,16,22-tetraazacyclotetracosane-2,5,8,11,14,17,20,23-octane(emodepside): In a 10 L reactor purged and maintained under an inert nitrogen atmosphere, EMD-18B (234.2 g, 0.21 mol, 1.0 equivalent) in ethyl acetate (3.8 L, 16.0 V) solution, N,N-diisopropylethylamine (131.84 g, 1.02 5.0 equivalents of 5 mol of propylphosphonic acid anhydride were added at 20-25°C. After cooling to 10-20°C, 324.5 g of 0.51 mol of propylphosphonic anhydride (2.5 equivalents) was added dropwise with stirring at below 25°C. The mixture was then stirred at 20-25°C for at least 2.5 hours and monitored by HPLC until EMD-18B was less than 0.5%. The mixture was diluted with heptane (2.38 L) at 0-10°C and slowly quenched with HCl (aqueous solution, 1.0 N, 2.38 L). The organic phase was washed with HCl (aqueous solution, 1.0 N, 2.38 L) and NaHCO3 (saturated, 2.38 L x 2), concentrated at below 40°C until no more distillate spilled, and then replaced with EtOH (0.48 L x 2) at below 40°C. The residue was dissolved in EtOH (0.72 L) at 45–55°C, and seed crystals (0.1%, w / w) were added at 20–25°C. The mixture was stirred at 20–25°C for at least 16 hours and filtered. The filtered cake was rinsed with EtOH (0.24 L). The solid was dried under vacuum at 40±5°C. The crude product was recrystallized with ethyl acetate. As a result, (3S,6R,9S,12R,15S,18R,21S,24R)-3,9,15,21-tetraisobutyl-4,6,10,16,18,22-hexamethyl-12,24-bis(4-morpholinobenzyl)-1,7,13,19-tetraoxa-4,10,16,22-tetraazacyclotetracosane-2,5,8,11,14,17,20,23-octane(emodepside) 118.24 g (51.3%) was obtained as a white solid; MS(ES,m / z):1120(M+H); 1H NMR(DMSO-d6,300 MHz)7.16(d,J=8.7 Hz,4H),6.87(d,J=8.2 Hz,4H),5.68(q,J=8.0 Hz,1H),5.51-4.04(m,7H),3.74-3.71(m,8H),3.08-3.04(m,8H),2.99-2.96(m,4H),2.90-2.88(m,4H), 2.83-2.82(m,4H),2.78(s,2H),2.70(s,2H),1.78-1.38(m,8H),1.31-1.14(m,6H),0.97-0.69(m,28H).

[0092] 3. Preparation of PF1022A (Method 1; refer to the reaction scheme in Figure 3 and the NMR spectrum in Figure 4) 3-1. Synthesis Procedure N-Fmoc-N-MeLeu-D-Lac-O-TAGa [ka] To a stirred solution of HO-TAGa (1.69 g, 1.85 mmol) in CH2Cl2 (37 mL), Unit 1 (12.0 mL, 2.40 mmol), 4-dimethylaminopyridine (12 mg, 93.0 μmol), and a 0.2 M toluene solution of N,N'-dicyclohexylcarbodiimide (1.30 g, 2.78 mmol) were added at room temperature under a N2 atmosphere. After stirring for 2 hours, the reaction mixture was cooled to -5°C and MeOH (185 mL) was added. The resulting heterogeneous solution was stirred further at -5°C for 15 minutes, the colorless precipitate was filtered, and washed with additional MeOH (500 mL) to obtain N-Fmoc-N-MeLeu-D-Lac-O-TAGa (2.46 g, 100%) as a colorless powder. mp: 44~45℃ [α] D 24 :-10.2(c 1.0, CHCl3) 1H-NMR(500 MHz,CDCl3)δ:7.78-7.74(complex m,2H),7.60-7.56(complex m,2H),7.39(m,2H),7.29(m,2H),6.50(s,4 / 3H),6.48(s,2 / 3H),5.12-5.00(complex-m,4H),4.67(dd,J=6.3,9.7 Hz,3 / 10H),4.58(dd,J=6.3 Hz,10.9 Hz,4 / 10H),4.50(dd,J=6.9 Hz,10.3 Hz,6 / 10H),4.38-4.34(complex m,9 / 10H),4.30(m,5 / 10H),4.23(t,J=6.3 Hz,3 / 10H),3.93(m,6H),2.86(s,2H),2.83(s 1H),1.76(m,6H),1.64-1.42(complex m,9H),1.31-1.14(complex m,87H),0.96-0.87(complex m,14H),0.78(d,J=6.9 Hz,1H). HRMS (FAB, NBA Matrix) m / z: 1334.0748 (M + , C 86 H 143 Calculated value for NO9: 1334.0763)

[0093] N-MeLeu-D-Lac-O-TAGa(1) [ka] Following the procedure described for the general deprotection of Fmoc, N-Fmoc-N-MeLeu-D-Lac-O-TAGa (1.00 g, 0.749 mmol) was converted to 1 (816 mg, 98%) as a colorless powder. 1H-NMR(500 MHz,CDCl3)δ:6.50(s,2H),5.16(q,J=6.9 Hz,1H),5.08(d,J=12.0 Hz,1H),5.04(d,J=12.0 Hz,1H),3.93(m,6H),3.30(t,J=7.5 Hz,1H),2.37(s,3H),1.75(m,6H),1.53-1.42(complex m,10H),1.32-1.25(complex m,86H),0.93-0.86(complex m,15H). 13 C-NMR(125 MHz, CDCl3)δ:170.6,153.3,138.4,130.2,106.4,73.5,69.2,68.9,67.6,61.4,42. 2,34.5,32.0,30.4,29.8(x2),29.5(x2),26.2,25.0,22.8,22.6,22.5,17.1,14.2. HRMS (FAB, NBA Matrix) m / z: 1113.0151 [(M+H) + , C 71 H 134 Calculated value for NO7: 1113.0160]

[0094] N-Fmoc-N-MeLeu-D-PhLac-N-MeLeu-D-LacO-TAGa(2) [ka] To a stirred solution of 1 (800 mg, 0.719 mmol) in CH2Cl2 (25 mL), unit 2 (426 mg, 0.827 mmol), N,N-diisopropylethylamine (0.429 mL, 2.52 mmol), and PyBroP (496 mg, 1.222 mmol) were added at room temperature. After stirring for 88 hours, the reaction mixture was crystallized according to the procedure described in the synthesis of N-Fmoc-N-MeLeu-D-Lac-O-TAGa to obtain 2 (1.11 g, 96%) as a colorless powder. 1H-NMR(500 MHz,CDCl3)δ:7.76-7.72(complex m,2H),7.62-7.40(complex m,2H),7.39-7.17(complex m,9H),6.47(m,2H),5.47-5.27(complex m,2H),5.15-4.97(complex m,4H),4.69-4.13(complex m,3H),3.92(complex m,6H),3.07(m,2H),2.85(complex,6H),1.80-1.25(complex m,105H),1.02-0.72(complex m,21H). HRMS (FAB, NBA Matrix) m / z: 1632.2163 [(M+Na) + , C 102 H 164 N2O 12 [Calculated value of Na: 1632.2182]

[0095] N-MeLeu-D-PhLac-N-MeLeu-D-Lac-OH(3) [ka] Following the procedure described for the general TAGa cleavage, 2 (614 mg, 0.381 mmol) was converted to 3 (261 mg, 95%) as yellow oil, which was used in the next reaction without further purification.

[0096] N-MeLeu-D-PhLac-N-MeLeu-D-Lac-O-TAGa(4) [ka] Following the procedure described for the general Fmoc deprotection, 2 (472 mg, 0.293 mmol) was converted to 4 (404 mg, 100%) as a colorless powder. 1H-NMR(500 MHz, CDCl3)δ:7.27(m,5H),6.49(s,2H),5.50(dd,J=5.7,8.6 Hz,1H),5.32(dd,J=4.6,10.9 Hz,1H),5.09-5.00(complex m,3H),3.93(m,6H),3.28(t,J=6.9 Hz,1H),3.09(m,2H),2.92(s,3H),2.27(s,3H),1.82-1.25(complex m,105H),0.89-0.77(complex m,21H). 13 C-NMR(125 MHz, CDCl3)δ:175.6,175.1,175.0,174.8,170.9,170.5(x2),170.4,169.8,169.7,166.7,165.1,153.3,138.5,138.3,135.8(x2 ),130.2,129.5,129.3,128.7,128.6,128.5(x2),127.2,107.0,106.9,105.4,78.1,73.5,71.9,71.6,69.4,69.2,68.9,67.7,67. 5,66.9,65.6,61.3,61.2,59.7,57.7,54.7,42.3,42.2,42.1,40.0,38.8,37.5,36.9,34.4,34.3,32.8,32.0,31.3,30.4,29.8(x2 ),29.6,29.5(x2),26.2,25.0,24.8,24.7,24.6,23.4,22.9,22.8,22.5(x2),22.4(x2),21.9,21.4,20.5,17.0,16.9,16.8,14.2. HRMS(FAB、NBAマトリックス)m / z:1388.1664[(M+H) + C 87 H 155 N2O 10 Calculated value:1388.1682]

[0097] N-Fmoc-N-MeLeu-D-PhLac-N-MeLeu-D-Lac-N-MeLeu-D-PhLac-N-MeLeu-D-Lac-O-TAGa(5)

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[0098] N-MeLeu-D-PhLac-N-MeLeu-D-Lac-N-MeLeu-D-PhLac-N-MeLeu-D-Lac-O-TAGa [ka] Following the procedure described for the general Fmoc deprotection, 5 (450 mg, 0.206 mmol) was converted to the corresponding amine (398 mg, 98%) as a colorless powder. 11H-NMR (500 MHz, CDCl3) δ: 7.25 (m, 10H), 6.48 (s, 2H), 5.57 - 4.99 (complex m, 9H), 3.93 (m, 6H), 3.26 - 2.78 (complex m, 14H), 2.24 (s, 3H), 1.80 - 1.25 (complex m, 114H), 0.90 - 0.78 (complex m, 33H). 13 13C-NMR (125 MHz, CDCl3) δ: 175.6, 175.2 (x2), 171.5, 171.1, 170.8 (x2), 170.6, 170.5 (x2), 170.3, 153.3, 138.5 (x2), 138.3, 136.1, 135.9, 135.5, 130.2, 130.1, 130.0, 129.8, 129.7, 129.6, 129.4 (x2), 128.8, 128.7, 128.6, 128.5, 127.4 (x2), 127.2, 127.1, 126.9, 126.8, 107.0 (x2), 106.9, 73.5, 72.5, 72.2, 71.7, 71.6, 69.8, 69.4, 69.2, 68.1, 68.0, 67.8, 67.5 (x2), 66.9, 61.4, 57.4, 55.2, 54.8, 54.6 (x2), 38.8, 37.7, 37.6, 37.4 (x2), 37.1 (x2), 37.0, 36.9, 34.7, 34.6, 32.2, 32.0, 31.9, 31.7, 31.6, 31.4, 31.3, 31.2, 30.4, 29.8 (x2), 29.6, 29.5 (x2), 26.2, 24.9, 24.8 (x2), 24.7, 24.6, 24.5, 23.5, 23.4, 23.3, 23.1 (x2), 23.0, 22.8, 22.7, 22.5, 22.4, 22.3, 22.0, 21.5, 21.4, 21.3 (x2), 20.5, 16.9, 16.8, 16.6, 14.2. HRMS (FAB, NBA matrix) m / z: 1862.4425 [(M + H) + 、C 113 H 193 N4O 16 の calculated value: 1862.4412]

[0099] N-MeLeu-D-PhLac-N-MeLeu-D-Lac-N-MeLeu-D-PhLac-N-MeLeu-D-Lac-OH [ka] Following the procedure described for the general TAGa cleavage, N-MeLeu-D-PhLac-N-MeLeu-D-Lac-N-MeLeu-D-PhLac-N-MeLeu-D-LacO-TAGa (380 mg, 0.204 mmol) was converted to the corresponding carboxylic acid as a yellow oil (198 mg, 98%), which was used in the next reaction without further purification. PF1022A [ka]

[0100] Reaction to a substrate at a concentration of 0.005 M The crude product from the previous reaction (90 mg, 0.0895 mmol) was dissolved in CH2Cl2 (18 mL, 0.005 M). N,N-diisopropylethylamine (76 μL, 4.48 mmol) and PyBOP (93 mg, 0.179 mmol) were added to the reaction mixture at room temperature. After stirring for 48 hours, the reaction mixture was quenched at 0°C with saturated NaHCO3 aqueous solution (18 mL), and the mixture was extracted with CHCl3 (20 mL x 2). The combined organic layer was washed with 10% NaHSO4 aqueous solution (60 mL) and saline solution (60 mL), dried over Na2SO4, filtered, and concentrated under vacuum. The crude product was purified by silica gel column chromatography (CHCl3:MeOH = 400:1~40:1) to obtain PF1022A (55 mg, 65%) as a colorless solid.

[0101] Reaction to a substrate at a concentration of 0.05 M Following the cyclization procedure described above, the crude product from the previous reaction (90 mg, 0.0895 mmol) was cyclized in CH2Cl2 (1.8 ml, 0.05 M) with N,N-diisopropylethylamine (76 μL, 4.48 mmol) and PyBOP (93 mg, 0.179 mmol) at room temperature for 48 hours to obtain PF1022A (40 mg, 49%) as a colorless solid. mp: 100~103℃ [α] 22 D -99.4℃ (C 0.06, MeOH) IR(KBr)ν:1743, 1666, 1466, 1412, 1265, 1188, 1126, 1080, 1026, 748, 701 1 H-NMR(500 MHz,CD3OD)δ:7.29(m,10H),5.82(t,J=7.5 Hz,1H),5.72(m,2H,rotamer),5.54(q,J=6.9 Hz,1H),5.44(dd,J=4.6,11.7 Hz,1H),5.40(dd,J=4.6,11.7 Hz,1H,rotamer),5.23(dd,J=4.6,11.7 Hz,1H),5.18(q,J=6.9 Hz,1H,rotamer),4.77(dd,J=3.4,11.2 Hz,1H),3.14(m,4H),3.00(s,5 / 2H,rotamer),2.91(m,7H,rotamer),2.82(s,5 / 2H,rotamer),1.84(m,1H),1.77-1.47(complex m,11H),1.39(d,J=6.3 Hz,3H),1.05(d,J=6.3 Hz,3H),0.98(d,J=6.9 Hz,3H),0.80(d,J=6.3 Hz,3H),0.95-0.82(complex m,18H). 13C-NMR(125 MHz, CD3OD)δ:174.4,173.5,173.1,173.1,172.3,172.0,171.0,170.7,136.4,136. 2,130.8,130.7,130.7,129.8,129.7,129.7,128.4,128.3,72.5,72.3,69.9,68.5, 58.6,55.7,55.5,55.4,39.0,38.9,38.6,38.6,37.8,37.3,32.0,31.3,31.1,30.0,26.2,26.1,25.5,25.2,23.9,23.7,23.7,23.6,21.7,21.6,21.4,21.1,17.5,17.2. HRMS(FAB、NBAマトリックス)m / z:971.5353[(M+Na) + C 52 H 76 N4O 12 Calculated value of Na1:971.5357] *Reference (J. Antibiot., 1992, 45, 692-697) mp:104~106℃ [α] 22 D -102℃ (c 0.1, MeOH) 1 H-NMR(400 MHz,CD3OD)δ:7.30-7.20(PhH,10H),5.80 and 5.75(C α H-PhLac, 1Hx2), 5.54 and 5.16 (C α H-Lac,1Hx2),5.43,5.42,5.22 and 4.78(C α H-Leu, 1Hx4), 3.22-3.15(C β H2-PhLac,2Hx2),3.00,2.90,2.88 and 2.80(N-Me-Leu,3Hx4),1.87-1.50(C β H2-Leu,2Hx4),1.40(C γ H-Leu, 1Hx4), 1.38(C β H3-Lac,3H), 1.02-0.75(C δ H3-Leu,6Hx4),0.88(C β H3-Lac,3H). 13 13C-NMR(100 MHz, CD3OD) δ: 174.4, 173.4, 172.4, 172.4, 172.1, 172.1, 171.0, 170.8, 136.5, 136.2, 130.7, 130.7, 130.7, 130.7, 129.7, 129.7, 129.7, 129.7, 128.3, 128.2, 72.5, 72.3, 69.9, 68.4, 58.6, 55.7, 55.5, 55.4, 39.0, 38.9, 38.6, 38.6, 37.9, 37.4, 32.0, 31.3, 31.1, 29.9, 26.2, 26.1, 25.6, 25.2, 23.6, 23.6, 23.5, 23.5, 21.7, 21.6, 21.4, 21.0, 17.5, 17.2.

[0102] 4. Preparation of PF1022A (Method 2; refer to the reaction scheme in Figure 5) 4-1. Synthesis procedure N-Fmoc-N-MeLeu-D-PhLac-O-TAGa

Chemical formula

[0103] N-MeLeu-D-PhLac-O-TAGa(6)

Chemical Structure

[0104] N-Fmoc-N-MeLeu-D-Lac-N-MeLeu-D-PhLacO-TAGa(7) [ka] To a stirred solution of 6 (358 mg, 0.301 mmol) in CH2Cl2 (6.0 ml), unit 1 (3.31 mL, 0.331 mmol), N,N-diisopropylethylamine (0.15 mL, 0.903 mmol), and a 0.1 M toluene solution of PyBroP (211 mg, 0.452 mmol) were added at room temperature. After stirring for 13 hours, the reaction mixture was crystallized according to the procedure described in the synthesis of N-Fmoc-N-MeLeu-D-Lac-O-TAGa to obtain 7 (468 mg, 97%) as a colorless powder. mp: 47~48℃ [α] D 26 :-13.9(c 1.10, CHCl3) 1H-NMR(500 MHz, CDCl3)δ:7.75(m,2H),7.59(m,2H),7.39(m,2H),7.30-7.09(complex m,7H),6.44(m,2H),5.36-4.96(complex m,6H),4.71-4.24(complex m,3H),3.93(t,J=6.5 Hz,6H),3.14(m,2H),2.90-2.81(complex s,6H),1.80-1.25(complex m,105H),0.96-0.76(complex m,21H). 13 C-NMR(125 MHz, CDCl3)δ:171.6,171.2,171.1,170.9,169.3,157.0,153.3,144.3,143.9,141.4(x2),138.4,135.9,130.0,129.9,129.8,129.7,129. 6,129.5,129.3,128.7,128.5,128.4,127.7,127.6,127.0,125.3,12 5.1,125.0,124.9,120.0,107.5,107.2,74.1,74.0,73.9,73.8,73.5, 71.3,69.2,67.9,67.7,57.4,57.2,56.7,56.6(x2),54.5(x2),47.4( x2),47.3,37.6,37.3,37.2,37.1,37.0,32.0,31.1,30.7,30.6(x2),3 0.5,30.4,30.3,30.2,29.8(x2),29.6,29.5(x2),26.2,24.9,24.8,2 4.7,24.6,24.4,23.4,23.2,22.9,22.8,22.1,21.4,21.2,16.8,14.2. HRMS(FAB、NBAマトリックス)m / z:1609.2274(M + C 102 H 168 N2O 12 Calculated value: 1609.2284)

[0105] N-MeLeu-D-Lac-N-MeLeu-D-PhLacOH(8)

change

[0106] N-MeLeu-D-Lac-N-MeLeu-D-PhLacO-TAGa(9) [ka] Following the procedure described for the general procedure of Fmoc deprotection, 7 (204 mg, 0.127 mmol) was converted to 9 (176 mg, 100%) as a colorless powder. mp: 47~48℃ [α] D 26 :-2.1(c 1.1, CHCl3) 1 H-NMR(500 MHz,CDCl3)δ:7.19(m,5H),6.45(s,2H),5.45(q,J=6.9 Hz,1H),5.31(ddd,J=4.6,11.2,19.6 Hz,1H),5.22(dd,J=5.2,6.9 Hz,1H),5.03(m,2H),3.94(m,6H),3.33-2.95(complex m,3H),2.80(s,3H),2.38(s,3H),1.82-1.25(complex m,105H),0.96-0.75(complex m,21H). 13C-NMR(125 MHz, CDCl3)δ:175.0,171.1,171.0,169.2,153.3,130.0,129.8,129.7,129.3,128.7,128 .6,128.5,127.4,127.1,126.9,107.5,107.2,73.8,73.5,71.3,69.2,67.9,67.7,67.4,67 .3,61.2,57.5,54.6,42.3,42.2,37.3,37.2,37.0,34.6,32.0,31.1,30.4,29.8(x2),29. 6,29.5(x2),26.2,25.0,24.8,23.4,23.0,22.8,22.7,22.4,22.1,21.3,16.9,16.8,14.2. HRMS (FAB, NBA Matrix) m / z: 1388.1676 [(M+H) + , C 87 H 155 N2O 10 Calculated value: 1388.1682]

[0107] N-Fmoc-N-MeLeu-D-Lac-N-MeLeu-D-PhLac-N-MeLeu-D-Lac-N-MeLeu-D-PhLac-O-TAGa(10) [ka] To a stirred solution of 9 (155 mg, 96.2 μmol) in CH2Cl2 (1.9 mL), 8 (103 mg, 0.144 mmol), N,N-diisopropylethylamine (73 μL, 0.434 mmol), and PyBroP (112 mg, 0.241 mmol) were added at room temperature. After stirring for 66 hours, the reaction mixture was crystallized according to the procedure described in the synthesis of N-Fmoc-N-MeLeu-D-Lac-O-TAGa to obtain 10 (201 mg, 100%) as a colorless powder. mp: 47~48℃ [α] D 27 :-27.2(c 1.1, CHCl3) 1H-NMR(500 MHz,CDCl3)δ:7.75(m,2H),7.59(m,2H),7.38(m,2H),7.30-7.12(complex m,12H),6.44(m,2H),5.43-4.96(complex m,10H),4.69-4.22(complex m,3H),3.93(m,6H),3.24-2.69(complex m,16H),1.80-1.25(complex m,114H),0.95-0.75(complex m,33H). 13 C-NMR(125 MHz,CDCl3)δ:174.1,171.5,171.4,171.3,171.2,171.1(x2),170.9,170.8,170.7,170.6,170.5,170.4,170.0,169.2,157.0,156.5,153.3,144.3,144.2,144.0(x2),141.4(x2),138.6,138.4,136.1,136.0,135.9(x2),135.7,135.6,135.3,130.0,129.8,129.7,129.6,129.5,129.3,128.8,128.7,128.6,128.4,127.7,127.4,127.1(x2),127.0.125.3,125.2,125.1,125.0,120.0,107.5,107.2,107.1,73.9,73.5,72.5,71.3,69.2,68.0,67.7,57.5,56.6(x2),55.1,55.0,54.9,54.7,54.4,47.4,47.3,40.9,40.6,38.7,37.6,37.3,37.2,37.1,32.0,31.9,31.7,31.5,31.3,31.2,31.0,30.7,30.6,30.4,30.3,29.8(x2),29.6,29.5(x2),25.0,24.9,24.8,24.7,24.5,24.4(x2),23.4,23.3,23.1,23.0,22.9,22.8,22.3,22.1,22.0,21.3,21.2,16.8,16.6,16.5,14.2. HRMS(FAB、NBAマトリックス)m / z:2106.4910[(M+Na) + 、C 128 H 202N4O 18 [Calculated value of Na: 2106.4912]

[0108] N-MeLeu-D-Lac-N-MeLeu-D-PhLac-N-MeLeu-D-Lac-N-MeLeu-D-PhLac-O-TAGa [ka] Following the procedure described for the general procedure of Fmoc deprotection, 10 (181 mg, 86.8 μmol) was converted to the corresponding amine (163 mg, 100%) as a colorless powder. mp: 47~48℃ [α] D 27 :-18.7(c 1.3, CHCl3) 1 H-NMR(500 MHz,CDCl3)δ:7.20(m,10H),6.44(s,2H),5.50-4.95(complex m,9H),3.93(m,6H),3.31-2.72(complex m,14H),2.36(m,3H),1.80-1.25(complex m,114H),0.99-0.81(complex m,33H). 13C-NMR (125 MHz, CDCl3)δ:171.4,171.2,171.0,170.9,170.6,170.4,153.3,138.3,13 6.1,136.0,135.9,135.8,130.0,129.8(x2),129.7,129.6(x3),129.5,129 .4,129.2,129.2(x2),128.8,128.7,128.6,128.4,127.4,127.3,127.2(x2 ),127.1,127.0,107.5,107.2,107.1,73.9,73.5,72.3,72.2,71.3,69.2,6 8.0(x2),67.7,67.4,61.3,55.2,55.0,54.8,54.5,54.4,42.4,40.6,38.7 ,37.7,37.2,37.1,34.7(x2),32.0,31.7,31.5,31.0,30.4,29.8(x2),29.6 ,29.5(x2),26.2,25.0,24.9(x2),24.7,23.5,23.4(x2),23.3,23.2,23.0, 22.8,22.4,22.1,21.9,21.3(x2),16.9,16.8(x3),16.7,16.6,16.5,14.2. HRMS (FAB, NBA Matrix) m / z: 1862.4462 [(M+H) + , C 113 H 193 N4O 16 Calculated value: 1862.4412]

[0109] N-MeLeu-D-Lac-N-MeLeu-D-PhLac-N-MeLeu-D-Lac-N-MeLeu-D-PhLac-OH [ka] Following the procedure described for the general TAGa cleavage, N-MeLeu-D-Lac-N-MeLeu-D-PhLac-N-MeLeu-D-Lac-N-MeLeu-D-PhLac-O-TAGa (143 mg, 0.768 mmol) was converted to the corresponding carboxylic acid (78 mg, 100%) as a yellow oil, which was used in the next reaction without further purification.

[0110] PF1022A [ka] The crude product from the previous reaction (78 mg, 0.0777 mmol) was dissolved in CH2Cl2 (16 mL, 0.005 M). N,N-diisopropylethylamine (66 μL, 0.389 mmol) and PyBOP (81 mg, 0.155 mmol) were added to the reaction mixture at room temperature. After stirring for 48 hours, the reaction mixture was quenched at 0°C with saturated NaHCO3 aqueous solution (16 mL), and the mixture was extracted with CHCl3 (20 mL x 2). The combined organic layers were washed with 10% NaHSO4 aqueous solution (60 mL) and saline solution (60 mL), dried over Na2SO4, filtered, and concentrated under vacuum. The crude product was purified by silica gel column chromatography (CHCl3:MeOH = 400:1~40:1) to obtain PF1022A (48 mg, 65%) as a colorless solid.

[0111] All physical data of the synthesized PF1022A matched the data of the genuine PF1022A. 5. Preparation of emodepside (Method 1; refer to the reaction scheme in Figure 6, the NMR spectrum in Figure 7, and the LC-UV spectrum in Figure 8)

[0112] 5-1. Synthesis Procedure N-Fmoc-N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-O-TAGa(11) [ka] To a stirred solution of 1 (259 mg, 0.233 mmol) in CH2Cl2 (4.7 mL), a 0.2 M toluene solution of unit 3 (0.122 mL, 0.244 mmol), N,N-diisopropylethylamine (0.12 mL, 0.698 mmol), and PyBroP (163 mg, 0.349 mmol) was added at room temperature. After stirring for 40 hours, the reaction mixture was crystallized according to the procedure described in the synthesis of N-Fmoc-N-MeLeu-D-Lac-O-TAGa to obtain 11 (395 mg, 100%) as a colorless powder. 1 H-NMR(500 MHz,CDCl3)δ:7.74(m,2H),7.57(m,2H),7.42-7.27(complex m,4H),7.05(m,2H),6.70(d,J=8.6 Hz,2H),6.48(s,2H),5.40(dd,J=6.9,8.4 Hz,3 / 10H,rotamer),5.35-5.27(complex m,17 / 10H),5.10-4.97(complex m,4H),4.72-4.12(complex m,3H),3.93(m,6H),3.75(m,4H),3.00-2.79(complex m,12H),1.80-1.60(complex m,9H),1.49-1.42(complex m,9H),1.27(complex m,87H),0.93-0.80(complex m,21H). HRMS (FAB, NBA Matrix) m / z: 1717.2701 [(M+Na) + , C 106 H 171 N3O 13 Calculated value of Na: 1717.2710]

[0113] N-Fmoc-N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-OH(12) [ka] Following the procedure described for general TAGa cleavage, 11 (210 mg, 0.124 mmol) was converted to 12 (100 mg, approximately 0.124 mmol) as brown oil. This crude product was used in the next reaction without further purification.

[0114] N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-O-TAGa(13) [ka] Following the procedure described for the general procedure of Fmoc deprotection, 11 (158 mg, 0.0934 mmol) was converted to 13 (138 mg, 100%) as a colorless powder. 1 H-NMR(500 MHz,CDCl3)δ:7.15(m,2H),6.83(d,J=8.6 Hz,2H),6.49(s,2H),5.47(dd,J=6.9,8.0 Hz,1H),5.32(dd,J=4.9,11.2 Hz,1H),5.09-5.00(complex m,3H),3.94(m,6H),3.85(m,4H),3.27(t,J=6.9 Hz,1H),3.13-2.79(complex m,9H),2.29(s,3H),1.81-1.25(complex m,105H),1.01-0.78(complex m,21H). HRMS (FAB, NBA Matrix) m / z: 1473.2214 [(M+H) + , C 91 H 162 N3O 11 Calculated value: 1473.2209]

[0115] N-Fmoc-N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-O-TAGa(14) [ka] To a stirred solution of 13 (138 mg, 0.934 mmol) in CH2Cl2 (1.9 mL), 12 (100 mg, approximately 0.124 mmol), N,N-diisopropylethylamine (48 μL, 0.280 mmol), and PyBroP (65 mg, 0.140 mmol) were added at room temperature. After stirring for 18 hours, the reaction mixture was crystallized according to the procedure described in the synthesis of N-Fmoc-N-MeLeu-D-Lac-O-TAGa to obtain 14 (211 mg, 100%) as a colorless powder. 1H-NMR(500 MHz,CDCl3)δ:7.75(m,2H),7.58(m,2H),7.42-7.28(complex m,4H),7.09(m,4H),6.77(m,4H),6.48(s,2H),5.45-5.15(complex m,6H),5.06-4.97(complex m,4H),4.73-4.12(complex m,3H),3.95-3.73(complex m,14H),3.15-2.73(complex m,24H),1.80-1.25(complex m,114H),0.96-0.77(complex m,33H). HRMS (FAB, NBA Matrix) m / z: 2276.5962 [(M+Na) + , C 136 H 216 N6O 20 [Calculated value of Na: 2276.5967]

[0116] N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-O-TAGa [ka] Following the procedure described for the general procedure of Fmoc deprotection, 14 (211 mg, 0.0934 mmol) was converted to the corresponding amine (178 mg, 94%) as a colorless powder. 1 H-NMR(500 MHz,CDCl3)δ:7.15(m,4H),6.82(m,4H),6.49(s,2H),5.14(t,J=7.45 Hz,1H),5.44-5.09(complex m,5H),5.07-5.00(complex m,3H),3.93(m,6H),3.85(m,8H),3.32(m,1H),3.17-2.74(complex m,21H),2.33(s,3H),1.81-1.64(complex m,12H),1.55-1.25(complex m,102H),1.03-0.77(complex m,33H). HR-MS(FAB, NBA Matrix + NaI)m / z:2032.5488[(M+H) + , C 121 H 207 N6O 18 [Calculated value: 2032.5467]

[0117] N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-OH [ka] Following the procedure described for the general TAGa cleavage, N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-N-MeLeu-D-LacO-TAGa (178 mg, 0.0876 mmol) was converted to the corresponding carboxylic acid (approximately 0.0876 mmol) as crude oil, which was used in the next reaction without further purification.

[0118] Emodepside (highly diluted using PyBOP) [ka] To the crude carboxylic acid product (approximately 0.0876 mmol) in CH2Cl2 (18 mL, 0.005 M), N,N-diisopropylethylamine (0.10 mL, 0.613 mmol) and PyBOP (91 mg, 0.175 mmol) were added at room temperature. After stirring for 19 hours, the reaction mixture was quenched at 0°C with saturated NaHCO3 aqueous solution (18 mL), and the mixture was extracted with CHCl3 (20 mL x 3). The combined organic layer was washed with 10% NaHSO4 aqueous solution (60 mL) and saline solution (60 mL), dried over Na2SO4, filtered, and concentrated under vacuum. The crude product was purified by silica gel column chromatography (CHCl3:MeOH = 400:1~100:1) to obtain emodepside (45 mg, 46% in 2 steps) as a pale yellow solid. mp: 98~103℃ [α] D24 :-40.3 (c 0.67, CHCl3) IR (neat) ν max : 2954, 2862, 1743, 1659, 1520, 1458, 1412, 1265, 1234, 1188, 1119, 1072, 1026, 926, 810. 1 1H-NMR (500 MHz, CDCl3) δ: 7.14 - 7.11 (complex m, 4H), 6.83 - 6.79 (complex m), 5.64 - 5.54 (complex m), 5.52 - 5.43, 5.43 - 5.39, 5.33, 5.18, 5.06 and 4.46 (5 m, 6H), 3.85 (apparently t, 8H), 3.15 - 3.04 (complex m, 8H), 3.04 - 2.92 (complex m, 4H), 3.00, 2.82, 2.79, 2.72 and 2.71 (5 s, 12H), 1.82 - 1.24 (complex m, 18H), 1.03 - 0.79 (complex m, 30H). 13 13C-NMR (125 MHz, CDCl3) δ: 171.6, 171.2, 171.0, 170.9, 170.3, 170.2, 170.1, 169.8, 169.7, 150.2, 130.4, 130.2, 115.7, 115.6, 71.2, 70.8, 68.5, 66.8, 66.7, 57.1, 54.0, 53.9, 49.3, 49.2, 38.0, 37.5, 37.1, 36.9, 36.8, 36.7, 36.6, 36.1, 31.1, 30.6, 30.4, 29.7, 29.6, 29.3, 25.0, 24.8, 24.6, 24.5, 24.5, 24.1, 23.6, 23.5, 23.4, 23.3, 23.3, 23.1, 22.6, 21.6, 21.5, 21.1, 21.1, 21.0, 20.8, 17.1, 15.7. HRMS (ESI) m / z: 1141.6404 [(M + Na) + , C 60 H 90 N6O 14 Na calculated value: 1141.6413] * Literature (Eur. J. Org. Chem., 2012, 1546 - 1553) 1 H-NMR(400 MHz,CDCl3)δ:7.17-7.09(m,4 H,Ar-H),6.86-6.77(m,4 H,Ar-H),5.67-5.54(m,2 H,CαH-Lac),5.53-5.38,5.34,5.19,5.08 and 4.47(5 m,6 H,CαH-Leu,CαH-morphPhLac),3.88-3.81(pseudo-t,8 H,OCH2morpholine),3.15-3.08(m,8 H,N-CH2-morpholine),3.07-2.85(m,4 H,CβH2-morphPhLac),3.00,2.83,2.80,2.74 and 2.73(5s,12 H,NCH3),1.83-1.20(m,18 H,CβH2-Leu,CγH-Leu,CH3-Lac),1.05-0.77(m,30 H,CδH3-Leu,CβH3-Lac). 13 C-NMR(100 MHz, CDCl3)δ:171.7,171.2,171.0,170.6,170.4,170.2,169.8,141.7,130.6,130.4,116.9,116.1,71.3,70.8,68.6,66.9,66.6,66.5, 57.1,54.0,49.9,38.1,37.5,37.2,36.7,36.2,31.2,30.5,29.4,24.9,24.7,24.2,23.6,23.5,23.5,23.4,21.2,21.1,20.9,17.1,15.8.

[0119] Emodepside (slow addition using T3P) [ka] TAGa cleavage: TFA (1.8 mL) was added to a stirred solution of a linear compound on TAG (179.0 mg, 0.088 mmol) in DCM (1.8 mL) at room temperature. After stirring at room temperature for 6 hours, the solution was concentrated under vacuum. The resulting mixture was dissolved in toluene (10 mL) and concentrated three times under reduced pressure to remove excess TFA. The crude residue was dissolved in CH2Cl2 (1.0 mL) and then recrystallized for the cleaved TAGa material by adding MeOH (8.0 mL) at room temperature. The precipitate was filtered through a Celite® pad and washed with MeOH (20 mL). The combined filtrate was concentrated under vacuum. 4 M HCl / dioxane (0.05 M relative to the product) was added to the resulting product, followed by dilution with toluene (10 mL) and concentration to obtain a product without TFA salts. To remove excess HCl from the crude product, the product was again dissolved in toluene (10 mL) and concentrated twice under reduced pressure.

[0120] Cyclization: To a stirred solution of T3P® (50% in Â, 110 μL, 0.187 mmol) in DIPEA (110 μL, 0.187 mmol), crude linear compound (0.088 mmol) in DCM (1.8 mL, 0.05 M, including washings) was added dropwise at room temperature over 2.5 hours. After stirring at room temperature for 20 hours, the reaction mixture was quenched with saturated NaHCO3 aqueous solution (3.0 mL) and extracted with CHCl3 (2.0 mL x 3). The combined organic layers were dried over sodium sulfate, filtered, and concentrated under vacuum. The crude residue was purified by silica gel column chromatography (CHCl3 / MeOH = 100 / 1) to obtain emodepside (86.4 mg, 88%) as amorphous material. Analytical data were identified using real samples.

[0121] 6. Preparation of emodepside (Method 2; see reaction scheme in Figure 9) N-Fmoc-N-MeLeu-D-morphPhLac-O-TAGa [ka] To a stirred solution of HO-TAGa (381 mg, 0.417 mmol) in CH2Cl2 (8.4 mL), a 0.2 M toluene solution of Unit 3 (2.71 mL, 0.542 mmol), 4-dimethylaminopyridine (2.5 mg, 20.8 μmol), and N,N'-dicyclohexylcarbodiimide (129 mg, 0.626 mmol) was added at room temperature under a N2 atmosphere. After stirring for 1 hour, the reaction mixture was crystallized according to the procedure described in the synthesis of N-Fmoc-N-MeLeu-D-Lac-O-TAGa to obtain N-Fmoc-N-MeLeu-D-morphPhLac-O-TAGa (628 mg, 100%) as a colorless powder. mp: 46~47℃ [α] D 27 = -3.1 (c 1.0, CHCl3) 1 H-NMR(500 MHz,CDCl3)δ:7.78(d,J=7.5 Hz,1H),7.74(d,J=7.5 Hz,1H),7.57(m,2H),7.39(m,2H),7.27(m,2H),6.98(d,J=8.6 Hz,1H),6.96(d,J=8.6 Hz,1H),6.67(m,2H),6.49(2 s,rotamer 4:3,2H),5.17(2 dd,rotamer,J=4.0 Hz,8.0 Hz,1H),5.08-4.98(complex m,3H),4.67-4.13(complex m,3H),3.93(m,6H),3.73(m,4H),3.08(dd,J=4.0 Hz,14.6 Hz,1H),3,07-2.95(complex m,5H),2.80(rotamer 4:3,3H),1.76(m,6H),1.64-1.53(complex m,3H),1.45(m,6H),1.28(complex m,84H),0.93-0.86(complex m,14H),0.76(d,J=6.3 Hz,1H). HRMS (FAB, NBA Matrix) m / z: 1495.1583 (M + , C 96 H 154 N2O 10 (Calculated value: 1495.1604)

[0122] N-MeLeu-D-morphPhLac-O-TAGa(15) [ka] Following the procedure described for the general deprotection of Fmoc, N-Fmoc-N-MeLeu-D-morphPhLac-O-TAGa (628 mg, 0.417 mmol) was converted to 15 (530 mg, 100%) as a colorless powder. mp: 49~50℃ [α] D 27 = +5.7(c 1.0, CHCl3) 1 H-NMR(500 MHz,CDCl3)δ:7.08(d,J=8.6 Hz,2H),6.80(d,J=8.6 Hz,2H),6.51(s,2H),5.24(dd,J=4.0,9.7 Hz,1H),5.07(q,J=12.0 Hz,2H),3.94(m,6H),3.85(m,4H),3.18(m,2H),3.10(m,4H),3.00(d,J=10.3,14.3 Hz,1H),2.21(s,3H),1.76(m,6H),1.47(m,6H),1.34-1.25(complex m,87H),0.88(t,J=6.9 Hz,9H),0.80(d,J=6.9 Hz, 3H), 0.79 (d, J = 6.9 Hz, 3H). HRMS (FAB, NBA Matrix) m / z: 1274.0986 [(M+H) + , C 81 H 145 Calculated value of N2O8: 1274.1001]

[0123] N-Fmoc-N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-O-TAGa(16) [ka] To a stirred solution of 15 (530 mg, 0.417 mmol) in CH2Cl2 (8.4 mL), a 0.2 M toluene solution of Unit 1 (0.22 mL, 0.44 mmol), N,N-diisopropylethylamine (0.212 mL, 1.25 mmol), and PyBroP (291 mg, 0.62 mmol) was added at room temperature. After stirring for 16 hours, the reaction mixture was crystallized according to the procedure described in the synthesis of N-Fmoc-N-MeLeu-D-Lac-O-TAGa to obtain 16 (670 mg, 95%) as a colorless powder. mp: 48~49℃ [α] D 27 = -12.4 (c 1.0, CHCl3) 1 H-NMR(500 MHz,CDCl3)δ:7.76(m,2H),7.61(m,2H),7.38(m,2H),7.30(m,2H),7.02(m,2H),6.74(m,2H),6.49(2 s,rotamer,2H),5.38-5.22(complex m,2H),5.15-4.98(complex m,4H),4.47(complex m,3H),3.94(m,6H),3.81(m,4H),3.12-2.78(complex m,12H),1.82-1.56(complex m,9H),1.53-1.40(complex m,8H),1.34-1.26(complex m,88H),0.98-0.75(complex m,21H). HRMS (FAB, NBA Matrix) m / z: 1694.2828 (M + , C 106 H 171 N3O 13 (Calculated value: 1694.2812)

[0124] N-Fmoc-N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-OH(17) [ka] Following the procedure described for the general TAGa cleavage, 16 (376 mg, 0.222 mmol) was converted to 17. In the case of this substrate, the reaction took longer than the general conditions for TAGa cleavage (approximately 1 hour). The reaction of 16 in 50% TFA / CH2Cl2 at room temperature required stirring for 8 hours to consume all the starting materials, yielding product 17 (178 mg, 0.222 mmol) as crude oil, which was used in the next reaction without further purification.

[0125] N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-O-TAGa(18) [ka] Following the procedure described for the general procedure of Fmoc deprotection, 16 (290 mg, 0.171 mmol) was converted to 18 (251 mg, 100%) as a colorless powder. mp: 43~45℃ [α] D 25 = -2.8 (c 1.0, CHCl3) 1 H-NMR(500 MHz,CDCl3)δ:7.07(d,J=8.6 Hz,2H),6.79(d,J=8.6 Hz,2H),6.49(s,2H),5.46(q,J=6.9 Hz,1H),5.33(dd,J=5.2,10.9 Hz,1H),5.17(dd,J=5.2,7.5 Hz,1H),5.06(m,2H),3.95(m,6H),3.84(m,4H),3.33(t,J=7.5 Hz,1H),3.11-3.06(complex m,6H),2.82(2 s,rotamer 4:1,3H),2.40(s,rotamer,3H),1.82-1.59(complex m,10H),1.52-1.43(complex m,8H),1.34-1.25(complex m,87H),0.97-0.86(complex m,21H). HRMS(FAB, NBA Matrix + NaI)m / z:1473.2222[(M+H) + , C 91 H162 N3O 11 Calculated value: 1473.2209]

[0126] N-Fmoc-N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-O-TAGa(19) [ka] To a stirred solution of 18 (251 mg, 0.171 mmol) in CH2Cl2 (3.4 mL), 17 (178 mg, 0.222 mmol), N,N-diisopropylethylamine (87 μL, 0.513 mmol), and PyBroP (120 mg, 0.257 mmol) were added at room temperature. After stirring for 46 hours, the reaction mixture was crystallized according to the procedure described in the synthesis of N-Fmoc-N-MeLeu-D-Lac-O-TAGa to obtain 19 (350 mg, 91%) as a colorless powder. mp: 50~52℃ [α] D 25 = -25.5 (c 1.0, CHCl3) 1 H-NMR(500 MHz,CDCl3)δ:7.75(m,2H),7.62(m,2H),7.38(m,2H),7.30(m,2H),7.06(m,4H),6.77(m,4H),6.49(2 s,rotamer,2H),5.42-4.98(complex m,10H),4.73-4.20(complex m,3H),3.94(m,6H),3.84(m,8H),3.17-2.66(complex m,24H),1.80-1.64(complex m,14H),1.46-1.25(complex m,100H),1.00-0.77(complex m,33H). HRMS (FAB, NBA Matrix) m / z: 2276.5940 [(M+Na) + , C 136 H 216 N6O 20 [Calculated value of Na: 2276.5967]

[0127] N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-O-TAGa [ka] Following the procedure described for the general procedure of Fmoc deprotection, 19 (114 mg, 0.0505 mmol) was converted to the corresponding amine (103 mg, 100%) as a colorless powder. mp: 45~47℃ [α] D 25 = -19.6 (c 1.0, CHCl3) 1 H-NMR(500 MHz,CDCl3)δ:7.76(m,4H),6.79(m,4H),6.48(s,2H),5.50-4.96(complex m,9H),3.94(m,6H),3.83(m,8H),3.30(m,1H),3.16-2.74(complex m,21H),2.38(m,3H),1.80-1.55(complex m,9H),1.47-1.24(complex m,105H),1.00-0.81(complex m,33H). HRMS (FAB, NBA Matrix) m / z: 2032.5468 [(M+H) + , C 121 H 207 N6O 18 [Calculated value: 2032.5467]

[0128] N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-OH [ka] Following the procedure described for the general TAGa cleavage, N-MeLeu-D-Lac-N-MeLeu-D-morphPhLac-N-MeLeu-D-Lac-N-MeLeu-D-morph-PhLac-O-TAGa (102 mg, 0.0502 mmol) was converted to the corresponding carboxylic acid. In the case of this substrate, the reaction took longer than the general conditions for TAGa cleavage (approximately 1 hour). The reaction at room temperature in 50% TFA / CH2Cl2 required stirring for 5 hours to consume all the starting materials, yielding the desired product (approximately 0.0502 mmol) as crude oil, which was used in the next reaction without further purification.

[0129] Emodepside [ka] To the crude carboxylic acid product (approximately 0.0502 mmol) in CH2Cl2 (10 mL, 0.005 M), N,N-diisopropylethylamine (60 μL, 0.351 mmol) and PyBOP (52 mg, 0.175 mmol) were added at room temperature. After stirring for 44 hours, the reaction mixture was quenched at 0°C with saturated NaHCO3 aqueous solution (10 mL), and the mixture was extracted with CHCl3 (20 ml x 3). The combined organic layer was washed with 10% NaHSO4 aqueous solution (60 ml) and saline solution (60 ml), dried over Na2SO4, filtered, and concentrated under vacuum. The crude product was purified by silica gel column chromatography (CHCl3:MeOH = 400:1 to 100:1) to obtain emodepside (25 mg, 44% in 2 steps) as a light brown solid.

[0130] All physical data for the synthetic emodepside matched those for the genuine compound.

[0131] Comparative Example 1 Similar to TAGa, but C 18 Instead of an alkyl chain, C 12 A tag (C12-TAG) containing the following was prepared and coupled with N-Fmoc-N-MeLeu-D-Lac-OH (unit 1) according to the following reaction scheme: [ka] Purification via silica gel chromatography was required after each reaction step. Compounds 2, C12-TAG, and 3 did not crystallize in methanol. This is in contrast to the synthetic procedure for N-Fmoc-N-MeLeu-D-Lac-O-TAGa (see Section 2.1 above), for example. The C12-TAG in this comparative example was not suitable for the intended tag-assisted synthesis, and further functionalization of compound 3 was not investigated.

[0132] Comparative Example 2 A commercially available C1-TAG was coupled with N-Fmoc-N-MeLeu-D-Lac-OH (unit 1) according to the following reaction scheme: [ka] Following the reaction steps, purification via column chromatography was required. Compounds C1-TAG and 6 did not crystallize in methanol. C1-TAG in this comparative example was not suitable for the intended tag-assisted synthesis, and further functionalization of compound 6 was not investigated.

Claims

1. A method for synthesizing a cyclic depsipeptide according to general formula (I) from a depsipeptide according to general formula (IIa): 【Chemistry 1】 (In the formula, Y is an amine protecting group and X is a carboxylic acid protecting group.) And, - The step of deprotecting an amine group protected by a Y group in the presence of an acid, thereby obtaining a deprotected amine group; - The step of deprotecting a carboxylic acid protected by an X group by hydrolysis, thereby obtaining a deprotected carboxylic acid group; - The steps of condensing the deprotected amine group and carboxylic acid group with a coupling agent to obtain a cyclic depsipeptide (I) and Includes, R2 and R8 are, independently of each other, hydrogen, linear or branched C1-C8 alkyl, linear or branched halogenated C1-C8 alkyl, hydroxy-C1-C6 alkyl, C1-C4-alkanoyloxy-C1-C6 alkyl, C1-C4-alkoxy-C1-C6 alkyl, aryl-C1-C4-alkyloxy-C1-C6 alkyl, mercapto-C1-C6 alkyl, C1-C4-alkylthio-C1-C6 alkyl, C1-C4-alkylsulfinyl-C1-C6 alkyl, C1-C4-alkylsulfonyl-C1-C6 alkyl, carboxy-C1-C6 alkyl, C1-C4-alkoxycarbonyl-C1-C6 alkyl, C1-C4 - Represents arylalkoxycarbonyl-C1-C6-alkyl, carbamoyl-C1-C6-alkyl, amino-C1-C6-alkyl, C1-C4-alkylamino-C1-C6-alkyl, C1-C4-dialkylamino-C1-C6-alkyl, guanidino-C1-C6-alkyl, C1-C4-alkoxycarbonylamino-C1-C6-alkyl, 9-fluorenylmethoxycarbonyl (Fmoc)amino-C1-C6-alkyl, C2-C8-alkenyl, C3-C7-cycloalkyl, C3-C7-cycloalkyl-C1-C4-alkyl, benzyl, substituted benzyl, phenyl, and optionally substituted phenyl-C1-C4-alkyl with halogens. x is 1, y is 1, R1, R4, R7 and R10 are methyl, R6 and R12 are methyl, R5 and R11 are independently linear or branched C1-C4 alkyl or linear or branched halogenated C1-C4 alkyl, and R3 and R9 are independently benzyl or p-morpholino-substituted benzyl. The coupling agent is T3P (registered trademark) (propylphosphonic anhydride, 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphospholinane-2,4,6-trioxide, PPACA). method.

2. X is unsubstituted - CH 2 - It is an aryl group, or X is selected from the following groups: benzyl (Bn), 4-methoxybenzyl (PMB), 3,4-dimethoxybenzyl (DPMB), 4-phenylbenzyl (PPB), 2-naphthylmethyl (Nap), and benzyloxymethyl acetal (BOM). The method according to claim 1.

3. The method according to claim 1 or 2, wherein Y is t-butyloxycarbonyl (BOC).

4. The depsipeptide according to the above general formula (IIa) is - In precursor (IV), the amine group protected by the PG2 group is deprotected in the presence of a base, thereby obtaining the deprotected amine group; - In precursor (III), the carboxylic acid protected by the PG3 group is deprotected in the presence of an acid, thereby obtaining the deprotected carboxylic acid group; - The deprotected amine group and carboxylic acid group are condensed to obtain a depsipeptide (IIa). By doing so, Obtained from precursors according to general formulas (IV) and (III): 【Chemistry 2】 (In the formula, R1 to R12, x and y have the meanings defined in claim 1, X has the meaning defined in claim 2, Y has the meaning defined in claim 3, PG2 is an amine protecting group, and PG3 is a carboxylic acid protecting group.) The method according to any one of claims 1 to 3.

5. The precursor according to the above general formula (IV) is - In precursor (VI), the amine group protected by the PG4 group is deprotected in the presence of a base, thereby obtaining the deprotected amine group; - The deprotected amine group of precursor (VI) and the carboxylic acid group of precursor (V) are condensed to obtain precursor (IV). By doing so, Obtained from precursors according to general formulas (VI) and (V): 【Transformation 3】 (In the formula, R7 to R12 and x have the meanings defined in claim 1, X has the meaning defined in claim 2, PG2 has the meaning defined in claim 4, and PG4 is an amine protecting group.) The method according to claim 4.

6. The precursor according to the above general formula (III) is - In precursor (VII), the amine group protected by the PG5 group is deprotected in the presence of a base, thereby obtaining the deprotected amine group; - The deprotected amine group of precursor (VII) and the carboxylic acid group of precursor (VIII) are condensed to obtain precursor (III). By doing so, Obtained from precursors according to general formulas (VIII) and (VII): 【Chemistry 4】 (In the formula, R1 to R6 and y have the meanings defined in claim 1, PG1 is an amine protecting group, PG3 has the meaning defined in claim 4, and PG5 is an amine protecting group.) The method according to claim 5.

7. The precursor according to general formula (VI) is obtained by esterification of the precursor according to general formula (IX) with X-LG: 【Transformation 5】 (In the formula, R10 to R12 have the meanings defined in claim 1, X has the meanings defined in claim 2, PG4 has the meanings defined in claim 5, and LG is a leaving group.) The method according to claim 6.

8. The precursor according to general formula (VII) is obtained by esterification of the precursor according to general formula (X) with PG3-OH: 【Transformation 6】 (In the formula, R4 to R6 and y have the meanings defined in claim 1, PG3 has the meaning defined in claim 4, and PG5 has the meaning defined in claim 6.) The method according to claim 7.

9. The method according to claim 4, 6, or 8, wherein PG3 is X.

10. The method according to any one of claims 4 to 6, wherein precursors (III) and (IV) are the same.

11. The method according to any one of claims 1 to 4, wherein R3 and R9 are the same, R2 and R8 are the same, and R5 and R11 are the same.

12. A method for synthesizing a cyclic depsipeptide according to general formula (I) from a depsipeptide according to general formula (IIb), wherein: 【Transformation 7】 (In the formula, PG1 is an amine protecting group, and TAG is a carboxylic acid protecting group.) - The step of deprotecting an amine group protected by a PG1 group in the presence of a base, thereby obtaining a deprotected amine group; - The step of deprotecting a carboxylic acid protected by a TAG group in the presence of an acid, thereby obtaining a deprotected carboxylic acid group; - The steps of condensing the deprotected amine group and carboxylic acid group with a coupling agent to obtain a cyclic depsipeptide (I) and Includes, The TAG group is partially Aryl-O-(CH 2 ) n It is characterized by containing - (Aryl represents the aromatic moiety, and n is 13 or greater), R2 and R8 are, independently of each other, hydrogen, linear or branched C1-C8 alkyl, linear or branched halogenated C1-C8 alkyl, hydroxy-C1-C6 alkyl, C1-C4-alkanoyloxy-C1-C6 alkyl, C1-C4-alkoxy-C1-C6 alkyl, aryl-C1-C4-alkyloxy-C1-C6 alkyl, mercapto-C1-C6 alkyl, C1-C4-alkylthio-C1-C6 alkyl, C1-C4-alkylsulfinyl-C1-C6 alkyl, C1-C4-alkylsulfonyl-C1-C6 alkyl, carboxy-C1-C6 alkyl, C1-C4-alkoxycarbonyl-C1-C6 alkyl, and C1-C4-arylalkoxycarbonyl -C1-C6-alkyl, carbamoyl-C1-C6-alkyl, amino-C1-C6-alkyl, C1-C4-alkylamino-C1-C6-alkyl, C1-C4-dialkylamino-C1-C6-alkyl, guanidino-C1-C6-alkyl, C1-C4-alkoxycarbonylamino-C1-C6-alkyl, tert-butoxycarbonylaminobutyl, 9-fluorenylmethoxycarbonyl (Fmoc)amino-C1-C6-alkyl, C2-C8-alkenyl, C3-C7-cycloalkyl, C3-C7-cycloalkyl-C1-C4-alkyl, benzyl, substituted benzyl, phenyl, phenyl-C1-C4-alkyl, hydroxyl, C1-C4-alkoxy or C1-C4-alkyl, x is 1, y is 1, R1, R4, R7 and R10 are methyl, R6 and R12 are methyl, R5 and R11 are independently linear or branched C1-C4 alkyl or linear or branched halogenated C1-C4 alkyl, and R3 and R9 are independently benzyl or p-morpholino-substituted benzyl. The coupling agent is T3P (registered trademark) (propylphosphonic anhydride, 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphospholinane-2,4,6-trioxide, PPACA). method.

13. PG1 is 9-fluorenylmethoxycarbonyl (Fmoc), t-butylcarbamate (Boc), benzylcarbamate (Z), acetamide, trifluoroacetamide, phthalimide, benzyl (Bn), triphenylmethyl (Tr), benzylidene, or p-toluenesulfonamide (Ts). TAG 【Transformation 8】 or 【Chemistry 9】 (In the formula, m is between 15 and 25, p is between 8 and 18, and q is between 15 and 25.) That is, The method according to claim 12.

14. The depsipeptide according to the above general formula (IIb) is - In precursor (IVb), the amine group protected by the PG2 group is deprotected in the presence of a base, thereby obtaining the deprotected amine group; - In precursor (IIIb), the carboxylic acid protected by the PG3 group is deprotected in the presence of an acid, thereby obtaining the deprotected carboxylic acid group; - The deprotected amine group and the carboxylic acid group are condensed to obtain the depsipeptide (IIb). By doing so, Obtained from precursors according to general formulas (IVb) and (IIIb): 【Chemistry 10】 (In the formula, R1 to R12, x and y have the meanings defined in claim 12, TAG has the meanings defined in claim 12 or 13, PG1 has the meanings defined in claim 6, and PG2 and PG3 have the meanings defined in claim 4.) The method according to claim 12 or 13.

15. The precursor according to the above general formula (IVb) is - In precursor (VIb), the amine group protected by the PG4 group is deprotected in the presence of a base, thereby obtaining the deprotected amine group; - The deprotected amine group of precursor (VIb) and the carboxylic acid group of precursor (Vb) are condensed to obtain precursor (IVb). By doing so, Obtained from precursors according to general formulas (VIb) and (Vb): 【Chemistry 11】 (In the formula, R7 to R12 and x have the meanings defined in claim 12, TAG has the meanings defined in claim 12 or 13, PG2 has the meanings defined in claim 4, and PG4 has the meanings defined in claim 5.) The method according to claim 14.

16. The precursor according to the above general formula (IIIb) is - In precursor (VIIb), the amine group protected by the PG5 group is deprotected in the presence of a base, thereby obtaining the deprotected amine group; - The deprotected amine group of precursor (VIIb) and the carboxylic acid group of precursor (VIIIb) are condensed to obtain precursor (IIIb). By doing so, Obtained from precursors by general formulas (VIIIb) and (VIIb): 【Chemistry 12】 (In the formula, R1 to R6 and y have the meanings defined in claim 12, PG1 has the meaning defined in claim 6, PG3 has the meaning defined in claim 4, and PG5 has the meaning defined in claim 6.) The method according to claim 15.

17. The precursor according to the above general formula (VIb) is obtained by esterification of the precursor according to the general formula (IXb) with TAG-OH: 【Chemistry 13】 (In the formula, R10 to R12 have the meanings defined in claim 12, TAG has the meanings defined in claim 12 or 13, and PG4 has the meanings defined in claim 5.) The method according to claim 15.

18. The precursor according to general formula (VIIb) is obtained by esterification of the precursor according to general formula (Xb) with PG3-OH: 【Chemistry 14】 (In the formula, R4 to R6 and y have the meanings defined in claim 12, PG3 has the meaning defined in claim 4, and PG5 has the meaning defined in claim 6.) The method according to claim 16.

19. The method according to claim 18, wherein PG3 is a TAG as defined in claim 12 or 13.

20. The method according to any one of claims 12 to 19, wherein at least one of the reaction steps that yield a TAG-containing molecule is followed by precipitation of the crude reaction product in methanol, thereby purifying the crude reaction product.

21. The method according to any one of claims 12 to 19, wherein at least one of the reaction steps in which the TAG-protected carboxylic acid group is deprotected is followed by precipitation of the cleaved TAG-OH in methanol and removal of the precipitate by filtration, thereby purifying the crude reaction product.

22. The aforementioned depsipeptide (II) is selected from one of the following general formulas: 【Chemistry 15】 【Chemistry 16】 【Chemistry 17】 [Chemistry 18] 【Chemistry 19】 【Chemistry 20】 (In the formula, A is a carboxylic acid protecting group and B is an amine protecting group.) The method according to any one of claims 1 to 21.

23. A method for synthesizing a cyclic depsipeptide according to general formula (I) from a depsipeptide according to general formula (IIc): 【Chemistry 21】 And, -E T (30) Value is 30 kcal mol -1 More than 43kcal mol -1 The following steps involve preparing a mixture of compound (IIc) and a base in a solvent, - The step of adding a coupling agent solution dropwise to a solvent to form a cyclic depsipeptide (I) Includes, R2 and R8 are, independently of each other, hydrogen, linear or branched C1-C8 alkyl, linear or branched halogenated C1-C8 alkyl, hydroxy-C1-C6 alkyl, C1-C4-alkanoyloxy-C1-C6 alkyl, C1-C4-alkoxy-C1-C6 alkyl, aryl-C1-C4-alkyloxy-C1-C6 alkyl, mercapto-C1-C6 alkyl, C1-C4-alkylthio-C1-C6 alkyl, C1-C4-alkylsulfinyl-C1-C6 alkyl, C1-C4-alkylsulfonyl-C1-C6 alkyl, carboxy-C1-C6 alkyl, C1-C4-alkoxycarbonyl-C1-C6 alkyl, C1-C4 - Represents arylalkoxycarbonyl-C1-C6-alkyl, carbamoyl-C1-C6-alkyl, amino-C1-C6-alkyl, C1-C4-alkylamino-C1-C6-alkyl, C1-C4-dialkylamino-C1-C6-alkyl, guanidino-C1-C6-alkyl, C1-C4-alkoxycarbonylamino-C1-C6-alkyl, 9-fluorenylmethoxycarbonyl (Fmoc)amino-C1-C6-alkyl, C2-C8-alkenyl, C3-C7-cycloalkyl, C3-C7-cycloalkyl-C1-C4-alkyl, benzyl, substituted benzyl, phenyl, and optionally substituted phenyl-C1-C4-alkyl with halogens. x is 1, y is 1, R1, R4, R7 and R10 are methyl, R6 and R12 are methyl, R5 and R11 are independently linear or branched C1-C4 alkyl or linear or branched halogenated C1-C4 alkyl, and R3 and R9 are independently benzyl or p-morpholino-substituted benzyl. The coupling agent is T3P (registered trademark) (propylphosphonic anhydride, 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphospholinane-2,4,6-trioxide, PPACA). method.

24. The E of the solvent T The value of (30) is 34 kcal / mol -1 or more and 39 kcal / mol -1 The method according to claim 23, wherein the value is 34 kcal / mol or more and 39 kcal / mol or less.

25. The method according to claim 23 or 24, wherein the ratio (moles:moles) of the base to compound (IIc) before the reaction is 2:1 or more and 10:1 or less.

26. The method according to any one of claims 23 to 25, wherein the ratio (moles:moles) of the coupling agent to compound (IIc) before the reaction is 1:1 or more and 5:1 or less.

27. The method according to any one of claims 23 to 26, wherein the temperature is maintained at 25°C or below during the addition of the coupling agent.

28. The method according to claim 1, wherein R2 and R8 each independently represent isobutyl.

29. The method according to claim 23, wherein R2 and R8 each independently represent isobutyl.

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