JPH07252295A - 6,7-dihydroxy-4-thiaheptanoic acid derivative - Google Patents

Abstract

PURPOSE:To obtain a low toxic new compound having excellent hyperleukocytotic activity, thus useful as a preventive/therapeutic agent for leukopenia developed by cancer radiotherapies or chemotherapies, or as a hematopoietic promoter or immunopotentiator in bone marrow transplantation. CONSTITUTION:The objective compound is expressed by formula I (R<1> and R<2> each is H or an aliphatic acyl; R<3> and R<4> each is H or a protecting group; X is an amino acid sequence made up of 1-10 amino acid residues; Y is an amino acid sequence bound to C-terminal made up of 1-5 amino acid sequences) or a salt thereof, e.g. (2R,6R)-2-glycylamino-6,7-bis (hexadecanoyloxy)-4- thiaheptanoyl-glutamyl-glycyl-D-glutamic acid. This compound of formula I can be obtained, for example, by condensation between a raw fragment of formula, H-X-OR<3> such as a compound of formula II and an amino group- protected raw fragment of formula III (R<5> is a protecting group for the amino group) such as 2-amino-6,7-bis(acyloxy-4-thiaheptanoic acid followed by further condensation of the resultant condensate with a compound of formula, R<4>-Y-OH.

Description

Detailed Description of the Invention

[0001]

The present invention relates to leukopenia or thrombocytopenia caused by various causes, diseases caused by a decrease in leukocytes or platelets, therapeutic bone marrow cells,
The present invention relates to a novel 6,7-dihydroxy-4-thiaheptanoic acid derivative useful as a therapeutic agent for a disease requiring an increase in white blood cells or platelets.

[0002]

2. Description of the Related Art Hoppe Sailors Tight Shrift Fur Physiologische Chemie (Hoppe-Se
yler's Zeitschrift fur Physiologische Chemie), 3
64, pages 593 to 606 (1983), a lipoprotein peptide derived from E. coli, which is represented by the following formula, is described.

[Chemical 2] Further, JP-A-4-46194 discloses a WS 1279A substance represented by the following formula.

[Chemical 3] These optically active isomers of 4-thiaheptanoic acid compounds are disclosed in JP-A-4-99796, Chemical Pharmaceuticals.
Bulletin (Chem. Pharm. Bull.), 39, 2590-2596.
Page (1991), Peptide Chemis
Try), 29, 361-366 (1991).
However, 2 of these 4-thiaheptanoic acid compounds
The amino group at position is substituted with C ₁₅ H ₃₁ CO-, and the structure is completely different from the amino acid sequence-substituted amino group bonded at the C-terminus of the compound of the present invention. Further, there is no description about the therapeutic action on thrombocytopenia. In the present specification, when an abbreviation is used for amino acids, peptides, etc., IUP is used.
AC-IUB Commission on Biochemicals
No Men Creature (Commission on Biochemical No
Menclature) or an abbreviation conventionally used in the art, and when an amino acid may have optical isomers, the L form is shown unless otherwise specified.

[0003]

When a chemotherapeutic method or radiation therapy for cancer is performed, the patient is caused to have severe leukopenia or severe thrombocytopenia. A major therapeutic problem is that leukopenia reduces resistance to infection and other diseases, and thrombocytopenia causes failure of the hemostatic mechanism, resulting in inability to obtain a sufficient therapeutic effect. Therefore, a drug that alleviates the suppression state of hematopoietic function caused by these treatment methods and promotes recovery of white blood cell count or platelet count is desired.
Further, there is a need for a drug capable of promoting the proliferation of transplanted bone marrow cells and promptly recovering the white blood cell count even when performing treatment by bone marrow transplantation. Furthermore, thrombocytopenia after bone marrow transplantation and autoimmune diseases with thrombocytopenia,
For example, this kind of drug can be used in the field of treatment of aplastic anemia, idiopathic thrombocytopenic purpura, etc.

[0004]

SUMMARY OF THE INVENTION In view of the above situation, the present inventors have repeatedly studied compounds having an action of increasing leukocytes and platelets, and as a result, a novel 4-thiaheptanoic acid derivative Promotes the growth of bone marrow cells,
It was found to increase peripheral leukocyte numbers and stimulate mouse bone marrow cells to promote proliferation and differentiation of megakaryocytes. Furthermore, the present inventors have completed the present invention as a result of continuing research based on these findings.

That is, the present invention provides (1) the general formula (I)

[Chemical 4] [In the formula, R ¹ and R ² are each hydrogen or an aliphatic acyl group, R ³ and R ⁴ are each hydrogen or a protective group,
X is an amino acid sequence consisting of 1 to 10 amino acid residues which may have a protecting group, and Y is a C-terminal consisting of 1 to 5 amino acid residues which may have a protecting group. Showing the amino acid sequence to be bound] or a salt thereof, (2) the compound according to (1) above, wherein the aliphatic acyl group has 2 to 30 carbon atoms, and (3) at least R ¹ and R ² . The compound according to (1) above, one of which is an aliphatic acyl group, and (4) 1 to 10, which may have a protecting group containing at least one residue of an amino acid having a hydrophilic group.
The compound according to (1) above, which is an amino acid sequence consisting of amino acid residues, (5) the compound according to (4) above, wherein the amino acid having a hydrophilic group is an acidic amino acid,
(6) The compound according to (4) above, wherein the amino acid having a hydrophilic group is a basic amino acid, (7) at least one of R ¹ and R ² is an aliphatic acyl group having 2 to 18 carbon atoms, X
Is an amino acid sequence consisting of 2 to 5 amino acid residues selected from glutamic acid, D-glutamic acid, glycine and lysine, which may have a protecting group, and Y is a glutamic acid which may have a protecting group. , Glycine, tyrosine,
The compound according to (1) above, which is an amino acid sequence consisting of 1 to 4 amino acid residues selected from lysine and β-alanine, (8) (2R, 6R) -2-glycylamino-
6,7-bis (hexadecanoyloxy) -4-thiaheptanoyl-glutamyl-glycyl-D-glutamic acid, the compound according to (1) above, (9) (2R, 6R).
2-glutamyl-glycylamino-6,7-bis (hexadecanoyloxy) -4-thiaheptanoyl-glutamyl-glycyl-D-glutamic acid (1).
The described compound, (10) (2R, 6R) -2-tyrosylamino-6,7-bis (hexadecanoyloxy) -4.
-Thiaheptanoyl-glutamyl-glycyl-D-glutamic acid, the compound according to (1), (11) (2)
R, 6R) -2-lysylamino-6,7-bis (hexadecanoyloxy) -4-thiaheptanoyl-glutamyl-glycyl-D-glutamic acid, the compound according to (1) above, (12) (2R, 6R). 2-glycylamino-6,7-bis (hexadecanoyloxy) -4-thiaheptanoyl-glycyl-glutamyl-glutamic acid, the compound described in (1) above, (13) the compound described in (1) above or a salt thereof. An immunopotentiating agent having a leukocytosis effect, which is contained, and (14) a therapeutic agent for thrombocytopenia, which comprises the compound according to (1) or a salt thereof.

With respect to the above general formula (I), examples of the aliphatic acyl group represented by R ¹ or R ² include an aliphatic acyl group derived from an aliphatic carboxylic acid. As the aliphatic acyl group, for example, an aliphatic acyl group having 1 to 34 carbon atoms, for example, a C _1-34 saturated aliphatic acyl group (eg,
Formyl, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, octanoyl, decanoyl, dodecanoyl, tridecanoyl, tetradecanoyl, pentadecanoyl, hexadecanoyl, heptadecanoyl, octadecanoyl, nonadecanoyl, nocosanoyl, tetracosanoyl, tetracosanoyl, tetracosanoyl, tetracosanoyl Noyl, methyl tridecanoyl, ethyl tridecanoyl, methyl tetradecanoyl, ethyl tetradecanoyl, methyl pentadecanoyl, ethyl pentadecanoyl, methyl hexadecanoyl, ethyl hexadecanoyl, methylheptadecanoyl, ethylheptadeca Noyl, methyl octadecanoyl, ethyl octadecanoyl, octacosanoyl, triacontanoyl, dotriacontanoyl, tetrato Akontanoiru, etc.), C _3-24
Unsaturated aliphatic acyl groups [eg, acryloyl, propioloyl, methacryloyl, crotonoyl, isocrotonoyl, myristoleoyl, oleoyl, palmitoreoyl, elideyl, (cis, cis) -9,12-octadecadienoyl, 9, 12,15-octadecatrienoyl, 9,11,13-octadecatrienoyl, 5,8,1
1,14-icosatetraenoyl, cis-15-tetracosaenoyl, etc.] and the like.

Preferably, an aliphatic acyl group having 2 to 30 carbon atoms, for example, a C _2-30 saturated aliphatic acyl group (eg, acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, octanoyl, decanoyl, dodecanoyl, tetra) Decanoyl, hexadecanoyl, ethyl dodecanoyl, methyl tridecanoyl, ethyl tridecanoyl, methyl tetradecanoyl, ethyl tetradecanoyl, methyl pentadecanoyl, ethyl pentadecanoyl, methyl hexadecanoyl, ethyl hexadecanoyl methyl hepta decanoyl, ethyl hepta decanoyl, methyl octadecanoyl, ethyl octadecanoyl, octadecanoyl, Nonakosanoiru, Tetorakosanoiru, Hekisakosanoiru, Okutakosanoiru, triacontanyl noil), the C _14-24 Saturated aliphatic acyl group [e.g.,
Myristoleoyl, oleoyl, palmitoreoyl, elideyl, (cis, cis) -9,12-octadecadienoyl, 9,12,15-octadecatrienoyl,
9,11,13-octadecatrienoyl, 5,8,11,
14-icosatetraenoyl, cis-15-tetracosaenoyl, etc.] and the like. Of these, aliphatic acyl groups having 2 to 18 carbon atoms, such as 10 to 1 carbon atoms
8 saturated aliphatic acyl groups (eg, decanoyl, dodecanoyl, tetradecanoyl, hexadecanoyl, octadecanoyl, etc.) or unsaturated aliphatic acyl groups having 14 to 18 carbon atoms [eg, myristoleoyl, oleoyl, Palmitoleoyl, (cis, cis) -9,12-octadecadienoyl, 9,12,15-octadecatrienoyl, etc.] are particularly preferable. At least one of R ¹ and R ² is preferably an aliphatic acyl group. Examples of such aliphatic acyl groups include saturated aliphatic acyl groups (eg, decanoyl, dodecanoyl, tetradecanoyl, hexadecanoyl, octadecanoyl, etc.), unsaturated aliphatic acyl groups [eg, myristoleoyl, Oleoyl, palmitoleoyl, (cis, cis) -9,12-octadecadienoyl, 9,12,15-octadecatrienoyl, etc.] are particularly preferable. Of these, the most preferred is a hexadecanoyl (palmitoyl) group.

In the above general formula (I), 1 to 10 optionally having a protecting group for the amino acid sequence represented by X.
The amino acid in each amino acid residue means a compound having an amino group and an acidic group (eg, carboxyl group, sulfo group) in the molecule. More specifically, for example, amino acids constituting proteins [eg, aliphatic monoaminomonocarboxylic acids such as glycine, alanine, valine, leucine and isoleucine, aliphatic oxyamino acids such as serine and threonine, aspartic acid, glutamic acid, etc. Monoaminodicarboxylic acids, monoaminodicarboxylic acid amides such as asparagine and glutamine, aromatic amino acids such as phenylalanine, tyrosine and tryptophan, iminocarboxylic acids such as proline and hydroxyproline, diaminomonocarboxylic acids such as arginine, lysine and histidine, methionine , Amino acids obtained from the natural world as microbial metabolites or animal and plant components [eg, L-α-aminobutyric acid, γ-aminobutyric acid, β-aminoisobutyric acid, β-ara] Nin, homoserine, α-methyl-D-serine, O-carbamyl-D
-Aserine, δ-hydroxy-γ-oxo-norvaline and other aliphatic monoaminomonocarboxylic acids, L-α-aminoadipic acid, L-β-aminoadipic acid, L-theanine, L-γ-methyleneglutamic acid, L -mono-aminodicarboxylic acids such as γ-methylglutamic acid, L-ornithine, β-lysine,
Diaminomonocarboxylic acids such as α, β-diaminopropionic acid, L-α, γ-diaminobutyric acid, diaminodicarboxylic acids such as diaminopimelic acid, sulfonic acid-containing monoaminomonocarboxylic acids such as cysteic acid, and aminosulfones such as taurine Acid, kynurenine, 3,4-dioxyphenyl-L
-Aromatic amino acids such as alanine, 2,3-dicarboxyaziridine, [S] -2-amino-3- (isoxazolin-5-one-
4-yl) -propionic acid, heterocyclic amino acids such as anticapsin, L-4-oxalysine, L-4-oxolysine, [3R,
5R] -3,6-diamino-5-hydroxyhexanoic acid and other carboxylic acids having two or more amino groups, lanthionine, S-
Sulfur-containing amino acids such as methyl-L-cysteine, pipecolic acid, azetidine-2-carboxylic acid, cyclic amino acids such as [1R, 2S] -2-aminocyclopentane-1-carboxylic acid, citrulline, alanosine, L-azaserine, etc. , Amino acids obtained by organic synthesis [eg, 6-aminohexanoic acid, 8-aminooctanoic acid, 12-
Aminododecanoic acid, 4-aminobenzoic acid, 4- (aminomethyl) benzoic acid, 4- (N- (carboxymethyl) aminomethyl) benzoic acid and the like) can be mentioned. The amino acid residue in X represents a divalent group derived from an amino acid having a bond to each of an amino group and an acidic group.

X is preferably an amino acid sequence consisting of 1 to 10 amino acid residues which may have a protecting group and which contains at least one amino acid residue having a hydrophilic group. Examples of the above-mentioned amino acid having a hydrophilic group include acidic amino acids, basic amino acids, and preferably acidic amino acids. Examples of acidic amino acids include one carboxyl group, one amino group, and other acidic functional groups (eg, carboxyl group, sulfo group, etc.)
A compound having one or more of Preferably 1
It is a compound having one amino group and two or more carboxyl groups. Preferable specific examples of acidic amino acids include aminodicarboxylic acids (eg, aspartic acid, glutamic acid, L-α-aminoadipic acid, L-β-aminoadipic acid, 2,3-dicarboxyaziridine, etc.). More preferred are α-aminodicarboxylic acids (eg, aspartic acid, glutamic acid, L-α-aminoadipic acid, etc.), and aspartic acid and glutamic acid are particularly preferred.

Examples of the basic amino acid include a carboxyl group, an amino group, and other basic functional groups [eg, amino group, basic nitrogen-containing heterocyclic group (eg, imidazolyl, indolyl, etc.), guanidino group, etc.] A compound having one or more of Preferably one amino group,
It is a compound having one carboxyl group and one or more of the above basic functional groups. Preferred specific examples of the basic amino acid include, for example, basic amino acids (eg, arginine, histidine, lysine, etc.) constituting proteins, basic amino acids obtained from the natural world as microbial metabolites or animal and plant components (eg, Diaminocarboxylic acid such as L-ornithine, β-lysine, α, β-diaminopropionic acid, L-α, γ-diaminobutyric acid, L-4-oxalysine, L-4-oxolysine, [3R, 5R] -3, 6-
Diamino-5-hydroxyhexanoic acid and the like). More preferably, a basic α-amino acid (eg, arginine, histidine, lysine, etc.) constituting a protein, a basic α-amino acid obtained from the natural world as a microbial metabolite or an animal or plant component (eg, L-ornithine, α,
β-diaminopropionic acid, L-α, γ-diaminobutyric acid, L-4-oxalysine, L-4-oxolysine, etc.). Particularly preferred are lysine, arginine and histidine.

1 in the amino acid sequence represented by X above
To 10 amino acid residues may have a protecting group used for protecting an amino group, a carboxyl group or a hydroxyl group in peptide synthesis, such as hydrolysis, hydrogenolysis, reduction, It is a protecting group that is eliminated by aminolysis or hydrazinolysis. Examples of the amino group-protecting group include urethane-type protecting groups (eg, benzyloxycarbonyl, t-butyloxycarbonyl, allyloxycarbonyl, t-amyloxycarbonyl, isobornyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2- Chlorbenzyloxycarbonyl, adamantyloxycarbonyl, 9-
Fluorenylmethyloxycarbonyl, 2,2,2-trichloroethyloxycarbonyl etc.), acyl type protecting groups (eg formyl, acetyl, propyl, trifluoroacetyl, chloroacetyl etc. substituents having 1 to 6 carbon atoms) A lower fatty acid residue which may have phthalyl,
Tosyl, 2-nitrosulfenyl, 4-methoxy-2-
Nitrosulfenyl, benzoyl etc.), aralkyl type protecting groups (eg trityl, benzhydryl, benzyl etc.) and the like. Of these, urethane type protecting groups are particularly preferable.

Carboxyl groups are protected by, for example, amidation, hydrazideation or esterification.
The amide group or hydrazide group is preferably substituted. The amide group is preferably substituted with, for example, a C7-19 aralkyl group which may be substituted with an alkoxy group (eg, 3,4-dimethoxybenzyl group or bis- (p-methoxyphenyl) -methyl group). . The hydrazide group is, for example, a halogen (eg, fluorine, chlorine, bromine, etc.) or a C1-12 alkyloxycarbonyl group optionally substituted by a C6-12 aryl group (eg, phenyl, p-biphenylyl, etc.) [eg, benzyloxy]. Carbonyl group, trichloroethyloxycarbonyl group, tert-butyloxycarbonyl group, 2- (p-biphenylyl) -isopropyloxycarbonyl group, etc.], halogenated C2-6 alkanoyl group (eg, trifluoroacetyl group, etc.), C7- It is preferably substituted with 19 aralkyl group (eg, trityl, benzhydryl, benzyl group, etc.) and the like. The carboxyl group is, for example, a lower alcohol which may be substituted by a lower alkoxy group or a halogen atom (eg,
Methanol, ethanol, cyanomethyl alcohol, benzoylmethyl alcohol, tert-butanol, etc.),
Alalkanols [eg, benzyl alcohol or benzhydrols optionally substituted by a lower alkyl group, a lower alkoxy group or a halogen atom (eg, benzhydrol, p-nitrobenzyl alcohol, p-
Methoxybenzyl alcohol, 2,4,6-trimethylbenzyl alcohol, etc.], and phenol and thiophenol optionally substituted with an electron-withdrawing substituent (eg, thiophenol, thiocresol, p-nitrothiophenol, 2 , 4,5- and 2,4,6-trichlorophenol, p-cyanophenol, p-methanesulfonylphenol, and N-hydroxyimides (eg, N
-Hydroxysuccinimide, N-hydroxyphthalimide, etc.), N-hydroxypiperidine, 8-hydroxyquinoline, etc. are preferred. A special protecting group for the carboxyl group which can be eliminated under neutral conditions is disclosed in the published German patent application 270,649.
And a hydrocarbyl-silyl-ethyl group described in No. 0 (eg, 2- (trimethylsilyl) -ethyl group, etc.).

Hydroxy groups can be protected, for example, by acylation or etherification. In the case of acylation, a group derived from carbonic acid (eg, benzyloxycarbonyl group or ethyloxycarbonyl group) is particularly suitable. Suitable groups for etherification are, for example, benzyl, tetrahydropyranyl or te
It is an rt-butyl group. Furthermore, for protection of the hydroxy group, Chem. Ber., Vol. 100 (1967),
The 2,2,2-trifluoro-1-tert-butyloxycarbonylaminoethyl group or 2,2,2-trifluoro-1-benzyloxycarbonylaminoethyl group described on pages 3838 to 3849 is suitable. . Most preferably, X represents an amino acid sequence having 2 to 5 amino acid residues selected from glutamic acid, D-glutamic acid, glycine and lysine, which may have a protecting group.

In the above general formula (I), the amino acids in 1 to 5 amino acid residues which may have a protecting group in the amino acid sequence bound at the C-terminal represented by Y are represented by X described above. It has the same meaning as the amino acid in 1 to 10 amino acid residues which may have a protecting group of the amino acid sequence. In this case, the protecting group has the same meaning as the protecting group for the amino acid sequence represented by X, which may have a protecting group for 1 to 10 amino acid residues. Of these, Y is preferably an amino acid sequence having 1 to 4 amino acid residues selected from glutamic acid, glycine, tyrosine, lysine, and β-alanine, which may have a protecting group. Particularly preferably, Y
Represents an amino acid sequence consisting of 1 to 2 amino acid residues selected from glutamic acid, glycine, tyrosine and lysine, which may have a protecting group. In the above general formula (I), the protective group represented by R ³ is preferably the above-mentioned protective group for a carboxyl group. In the above general formula (I), the protecting group represented by R ⁴ is
Preferably, the aforementioned amino-protecting group is used. In the general formula (I), when the amino acid residue may have optical isomers, any of the L-, D-, and DL- configurations can be adopted.

Next, a method for producing the above compound will be described. The protecting groups and reagents commonly used in the text below are represented by the following abbreviations. Fmoc: 9--fluorenylmethyloxycarbonyl Z: benzyloxycarbonyl Boc: t-butoxycarbonyl ^t Bu: t-butyl O ^t Bu: t-butoxy TFA: Trifluoroacetic acid TEA: Triethylamine DMF: N, N-dimethylformamide DCC: N, N'-dicyclohexylcarbodiimide BOP: Benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate DIC: N, N'-diisopropylcarbodiimide HONB: N-hydroxy-5-norbornene-2,
3-Dicarboximide HOBT: 1-Hydroxybenzotriazole DEPC: Diethyl cyanophosphate DCM: Dichloromethane THF: Tetrahydrofuran WSC: Water-soluble carbodiimide / hydrochloride DMAP: 4-Dimethylaminopyridine MeOH: Methanol R-: R-coordination S- : S-coordination

The compound serving as the basic skeleton of the compound in the present text is 2-amino-6,7-dihydroxy-4-thiaheptanoic acid represented by the following formula.

[Chemical 5] In the text, thiaheptanoyl is referred to as THT, and thiaheptanoic acid is referred to as THT-OH. In addition, n-hexadecanoyl is referred to as Pam and n-hexadecanoyloxy is referred to as PamO.

Among the compounds represented by the above general formula (I) or salts thereof, the compounds or salts thereof in which R ³ and R ⁴ represent a protecting group are converted into several fragments at any position of the peptide bond. It can be produced by dividing and sequentially condensing a fragment having a reactive carboxyl group and a fragment having a reactive amino group by a method known per se. As the condensation method, for example, a conventional means for peptide synthesis can be used. As a conventional method for peptide synthesis,
For example, either a liquid phase synthesis method or a solid phase synthesis method may be used. The means for synthesizing such a peptide may be according to any known method, for example, M. Bondaski (M. Bonda).
sky) and M. Ondetti, Peptide Synthesis, Interscience, New York, 1966; FM Finn and K. Hofman.
n), The Proteins, Volume 2,
H. Nenrath and Earl El
Edited by RL Hill, Academic Press,
New York, 1976: Nobuo Izumiya et al., "Basics and Experiments in Peptide Synthesis," Maruzen Co., Ltd. 1985; Haruaki Yajima,
Shunpei Sakakibara et al., Biochemistry Experiment Course 1, edited by The Biochemical Society of Japan, Tokyo Chemistry Doujin 1977; Shun Kimura et al., Continuing Biochemistry Experiment Course 2, edited by The Biochemical Society of Japan, Tokyo Chemistry Doujinshi 1987; J. M. Stewart (JM Stewart) and JD Young, Solid Phase Peptide Sy
nthesis), Pierce Chemical Company, Illinois, 1
It is manufactured by the method described in 984 or the like or a method similar thereto. Specific examples of the method include, for example, azide method, chloride method, acid anhydride method, mixed acid anhydride method,
DCC method, active ester method, method using Woodward reagent K, carbonyl imidazole method, redox method, N,
N'-dicyclohexylcarbodiimide (DCC) / N-
Hydroxy-5-norbornene-2,3-dicarboximide (HONB) method, N, N'-diisopropylcarbodiimide (DIC) / HONB method, water-soluble carbodiimide hydrochloride (WSC) / 1-hydroxybenzotriazole (HOBT) Method, a method using a benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate (BOP) reagent, a method using diethyl cyanophosphate (DEPC), and the like.

When these methods are used, examples of the compound having activated carboxyl group of the raw material include corresponding acid anhydrides, azides, active esters [eg alcohols (eg pentachlorophenol, 2, 4, 5-trichlorophenol, 2,4-dinitrophenol, cyanomethyl alcohol, p-nitrophenol, N-hydroxy-5-norbornene-2,3-dicarboximide, N-hydroxysuccinimide, N-hydroxyphthalimide, N -Hydroxybenztriazole)
And the like] and the like. Examples of the activated amino group of the raw material include the corresponding phosphoric acid amide.

The condensation reaction can be carried out in the presence of a solvent. The solvent can be appropriately selected from those known to be usable in the peptide condensation reaction. For example, anhydrous or hydrous formamide, dimethylformamide, N-
Amides such as methylpyrrolidone, sulfoxides such as dimethyl sulfoxide, aromatic amines such as pyridine, halogenated hydrocarbons such as chloroform and dichloromethane, ethers such as tetrahydrofuran and dioxane, nitriles such as acetonitrile, acetic acid Examples thereof include esters such as ethyl and ethyl formate, or a mixture thereof in an appropriate ratio. The reaction temperature is appropriately selected from the range known to be applicable to peptide bond formation reactions. Specifically, for example, usually about -2
It is appropriately selected from the range of 0 ° C to 40 ° C. The reaction time is
It is appropriately selected from the range known to be necessary for the peptide bond forming reaction. Specifically, for example, from a few minutes to 48
React for about an hour.

The above general formula (I) [in the formula, R ³ and R ⁴ are protecting groups, and other symbols have the same meanings as described above. ] When synthesizing the compound represented by or its salt, as a preferable specific example of a raw material fragment, for example, general formula (II) HX-OR ³ (II) [wherein, X and R ³ have the same meanings as described above] ] The compound or its salt represented by these is mentioned. Compound (II) or a salt thereof can be produced by appropriately using the conventional means for peptide synthesis described above. Specific preferred examples of the compound (II) include the compounds shown below. Compound number Structural formula HP-1 H-Gly-Gly-Gly-Glu (O ^t Bu) -O ^t Bu HP-2 H-Glu (O ^t Bu) -Gly-D-Glu (O ^t Bu) -O ^t Bu HP-3 H-Gly-D-Glu (O ^t Bu) -O ^t Bu HP-4 H-Gly-Glu (O ^t Bu) -Glu (O ^t Bu) -O ^t Bu HP-5 H-Glu (O ^t Bu) -Gly-Glu (O ^t Bu) -O ^t Bu HP-6 H-Gly-Lys (Boc) -Gly-O ^t Bu

Further, as another raw material fragment to be combined with the above compound (II), a compound represented by the general formula (IV)

[Chemical 6] [In the formula, R ⁵ represents an amino-protecting group, and R ¹ and R ²
Has the same meaning as defined above] or a salt thereof. The amino group-protecting group represented by R ⁵ in the above general formula (IV) has the same meaning as the above-mentioned amino group-protecting group.

The compound (IV) or a salt thereof has R ¹ and R ²
In the same group, a method known per se [eg, International Journal of Peptide a
nd Protein Research), 38, 1991, 54
Pages 5 to 554, Chemical and Pharmaceutical Bullet
in), Vol. 39, pp. 2590-2596].

The most general production route in the case where R ¹ and R ² are different groups will be described below. First, a glycerin derivative, for example, a monoacyl derivative such as glycidol is prepared. On the other hand, 2-amino in which the amino group and the carboxyl group are protected by reductively cleaving the disulfide bond of cystine in which the amino group and the carboxyl group are protected to give a cysteine derivative, and the above-mentioned monoacylglycerin derivative is added thereto. -6-hydroxy-7-
Acyloxy-4-thiaheptanoic acid can be obtained. In this reaction, using R-(+)-glycidol gives a (6R) -2-amino-6-hydroxy-7-acyloxy-4-thiaheptanoic acid derivative, S-
If (-)-glycidol is used, a (6S) -2-amino-6-hydroxy-7-acyloxy-4-thiaheptanoic acid derivative is obtained. By acylating the 6-position hydroxyl group of the 2-amino-6-hydroxy-7-acyloxy-4-thiaheptanoic acid derivative obtained by the above reaction by a conventional method, 2-amino-6, which has a different O-acyl group,
A 7-bis (acyloxy) -4-thiaheptanoic acid derivative can be obtained.

As a modification of the above-mentioned production method, a 2-amino-6,7-dihydroxy-4-thiaheptanoic acid derivative can be obtained by directly using glycidol or the like instead of the acylglycerin derivative. When converted to 2-amino- having the same O-acyl group
A 6,7-bis (acyloxy) -4-thiaheptanoic acid derivative can be easily obtained. 2-amino-obtained in this way
2-amino-6,7-bis (acyloxy) having an amino group protected by removing the protecting group of the carboxyl group of the 6,7-bis (acyloxy) -4-thiaheptanoic acid derivative by, for example, the method described below. -4-Thiaheptanoic acid can be produced. Specific preferred examples of the compound (IV) include 2-amino-protected with an amino group.
6,7-bis (acyloxy) -4-thiaheptanoic acid may be mentioned. More specific examples are the compounds shown below. Compound number Reference example number Structural formula FmocGC-1 7 (2R, 6R) -2-Fmoc amino-6,7-bis (PamO) -4-THT-OH

The compound (II) and the compound (IV) obtained as described above are condensed by the conventional means for peptide synthesis described above, and the protecting group for the amino group represented by R ⁵ is removed by the method described below. Protect and general formula (III)

[Chemical 7] [Wherein R ¹ and R ² have the same meanings as described above] or a salt thereof can be produced. Specific examples of the compound (III) include the following compounds. Compound Reference Example Number Number Structural formula HGCP-1 9 (2R, 6R) -2-Amino-6,7-bis (PamO) -4-THT-Gly-Gly-Gly- Glu-O ^t Bu HGCP-2 10 ( 2R, 6R) -2-Amino-6,7-bis (PamO) -4-THT-Glu (O ^t Bu) -Gly- D-Glu (O ^t Bu) -O ^t Bu HGCP-3 11 (2R, 6R) -2-Amino-6,7-bis (PamO) -4-THT-Gly- D-Glu (O ^t Bu) -O ^t Bu HGCP-4 12 (2R, 6R) -2-Amino-6, 7-bis (PamO) -4-THT-Gly-Glu (O ^t Bu)-Glu (O ^t Bu) -O ^t Bu HGCP-5 13 (2R, 6R) -2-amino-6,7-bis ( PamO) -4-THT-Glu (O ^t Bu) -Gly- Glu (O ^t Bu) -O ^t Bu HGCP-6 14 (2R, 6R) -2-Amino-6,7-bis (PamO) -4 -THT-Gly-Lys (Boc)-Gly-O ^t Bu

Of the compounds represented by the general formula (I) or salts thereof, compounds in which R ³ and R ⁴ represent a protecting group or salts thereof are the same as the compounds (III) or salts thereof and the general formula (V ) R ⁴ -Y-OH (V) [wherein R ⁴ and Y have the same meaning as defined above] or a salt thereof is subjected to a condensation reaction by appropriately using a conventional peptide synthesis method. It is obtained by attaching. Y
Is an amino acid sequence consisting of 2 to 5 amino acid residues which may have a protecting group, a peptide fragment obtained by appropriately dividing compound (V) or an amino acid can be obtained by appropriately using a conventional method for peptide synthesis. It can also be produced by sequential condensation with (III) or a salt thereof.

Compound (I) thus produced
Alternatively, by subjecting a compound or a salt thereof, in which R ³ and R ⁴ are protecting groups, to a deprotection reaction, a compound of the general formula (I)
R ³ and R ⁴ among the compounds represented by
A compound or a salt thereof in which is hydrogen can be produced. The deprotection group reaction can be carried out by a method known per se, for example, a method commonly used in peptide chemistry. [Nobuo Izumiya, Motonori Ohno, Tetsuo Kato, Tohiko Aoyagi, Synthetic Chemistry Series, Peptide Synthesis, published by Maruzen Co., Ltd., 1975] In a solvent that does not adversely affect the reaction,
It is carried out by contact with an acid. As the solvent, halogenated hydrocarbons (eg, dichloromethane, chloroform, 1,2-dichloroethane, etc.), alcohols (eg, methanol, ethanol, etc.), water and a mixture thereof in an appropriate ratio are used. As the acid, for example, haloacetic acid (eg, trifluoroacetic acid, etc.), hydrohalic acid (eg, hydrochloric acid, hydrobromic acid, etc.) and the like are used. The N-benzyloxycarbonyl group and the N-4-methoxybenzyloxycarbonyl group are removed by catalytic hydrogenation using, for example, a palladium catalyst (eg, palladium carbon, palladium / barium sulfate, palladium black, etc.) or a rhodium catalyst. It is advantageous. At that time, solvents known from literature such as cyclic ethers (eg, tetrahydrofuran), alcohols (eg, methanol, ethanol, etc.) may be used in combination with other inert solvents [eg, lower aliphatic acid amides (eg, dimethylformamide, etc.). ) Etc.] The N-9-fluorenylmethyloxycarbonyl group is advantageously removed by using organic amines such as diethylamine, piperidine, morpholine, 4-dimethylaminopyridine and dicyclohexylamine. The reaction is carried out in a solvent that does not adversely influence the reaction. Examples of the solvent include amides (eg, dimethylformamide, acetamide, etc.), alcohols (eg, methanol, ethanol, etc.), halogenated hydrocarbons (eg, dichloromethane, chloroform, 1,2-dichloroethane, etc.), or the like. Mixtures of these in appropriate proportions are used. The N-2,2,2-trichloroethyloxycarbonyl group is an organic carboxylic acid (eg, acetic acid,
It is advantageous to use a metal (eg, zinc, etc.) together with puionic acid (for example). The reaction is carried out in a solvent that does not adversely influence the reaction. As the solvent, the above organic carboxylic acids, alcohols (eg, methanol, ethanol, etc.), water, etc., or a mixture thereof in an appropriate ratio is used. The deprotection reaction of the acylated hydroxy group (deacylation reaction) is carried out by contacting with an acid in a solvent that does not adversely influence the reaction. As the solvent, halogenated hydrocarbons (eg, dichloromethane, chloroform, 1,2-dichloroethane, etc.), alcohols (eg, methanol, ethanol, etc.), water and a mixture thereof in an appropriate ratio are used. As the acid, for example, haloacetic acid (eg, trifluoroacetic acid, etc.), hydrohalic acid (eg, hydrochloric acid, hydrobromic acid, etc.) and the like are used. The O-benzyl group is advantageously removed by catalytic hydrogenation using for example a palladium catalyst (eg palladium on carbon, palladium / barium sulphate, palladium black etc.) or a rhodium catalyst. At that time, solvents known from literature such as cyclic ethers (eg, tetrahydrofuran), alcohols (eg, methanol, ethanol, etc.) may be used in combination with other inert solvents [eg, lower aliphatic acid amides (eg, dimethylformamide, etc.). ) Etc.] O-tetrahydropyranyl group or Ot
The ert-butyl group can be hydrolyzed with an acid as in the case of deacylation. The elimination of the carboxyl protecting group can be carried out by acid hydrolysis as in the above. Further, for example, a benzyl ester can be eliminated by catalytic hydrogenation in the same manner as in the case of eliminating the O-benzyl group described above. 2- (trimethylsilyl)
-Ethyl groups are allowed to act under neutral conditions, such as salts of hydrofluoric acid, for example salts of hydrofluoric acid, especially quaternary nitrogen bases (eg tetraethylammonium fluoride, etc.) in a suitable solvent. Can be desorbed by.

After completion of the reaction, the compound (I) or salt thereof produced in this manner is used for peptide separation means such as extraction, partitioning, reprecipitation, crystallization, recrystallization, various chromatography, high performance liquid chromatography. It is collected by. The compound (I) of the present invention can be converted into a salt with a base, particularly a pharmaceutically acceptable base, by a method known per se. Examples of the base include alkali metals such as sodium and potassium, alkaline earth metals such as calcium and magnesium, and organic bases such as tertiary amines such as triethylamine and pyridine. Furthermore, it can also be obtained as an acid addition salt, especially a pharmacologically acceptable acid addition salt. Examples of the acid include inorganic acids (eg, hydrochloric acid, sulfuric acid, phosphoric acid, etc.) and organic acids (eg, acetic acid, propionic acid, citric acid, tartaric acid, malic acid, oxalic acid, etc.). Compound (II) above
As the salts of to (V), the same salts as the salts of compound (I) are used.

The compound (I) or a salt thereof has the ability to remarkably improve the hematopoietic deficiency state, and can be used for radiotherapy or chemotherapy for cancer of mammals (eg, humans, monkeys, horses, cows, pigs, dogs, etc.). It can be used as a therapeutic or preventive agent for leukopenia caused by the above, as a hematopoietic agent at the time of bone marrow transplantation, as an immunopotentiator having a leukocytosis effect, and as a therapeutic agent for thrombocytopenia. Compound (I) or a salt thereof has low toxicity and can be safely used. When compound (I) or a salt thereof is administered to, for example, a human, it is mixed with itself or with an appropriate pharmacologically acceptable carrier, excipient, diluent, and orally or parenterally as a pharmaceutical composition. Can be safely administered to. The above pharmaceutical composition,
Oral preparations (eg, powders, granules, capsules, tablets), injections, infusions, external preparations (eg, nasal preparations, transdermal preparations, etc.), suppositories (eg, rectal suppositories, vaginal suppositories) ) And the like. These preparations can be produced by a method known per se which is generally used in the preparation process.

The compound (I) of the present invention or a salt thereof is a dispersant (eg, Tween 80 (manufactured by Atlas Powder Co., USA), HCO 60 (manufactured by Nikko Chemicals) polyethylene glycol, carboxymethyl cellulose, sodium alginate, etc.). , Preservatives (eg, methylparaben, propylparaben, benzyl alcohol, etc.), isotonicity agents (eg, sodium chloride, mannitol, sorbitol,
(Glucose etc.) or the like, or an aqueous injection, or dissolved, suspended or emulsified in olive oil, sesame oil, cottonseed oil, vegetable oil such as corn oil, propylene glycol or the like to form an oily injection, which can be made into an injection. In order to prepare a preparation for oral administration, the compound (I) of the present invention or a salt thereof can be prepared by, for example, an excipient (eg, lactose,
Sucrose, starch, etc.), disintegrant (eg, starch, calcium carbonate, etc.), binder (eg, starch, gum arabic, carboxymethylcellulose, polyvinylpyrrolidone, hydroxypropylcellulose, etc.) or lubricant (eg, talc, stearin) Magnesium acid, polyethylene glycol 6000, etc.) may be added to the mixture for compression molding, and if necessary, it may be coated by a method known per se for the purpose of taste masking, enteric coating or sustainability to prepare an oral administration preparation. it can. Examples of the coating agent include hydroxypropylmethyl cellulose, ethyl cellulose, hydroxymethyl cellulose, hydroxypropyl cellulose, polyoxyethylene glycol, Tween 80, Bluronic.
F 68, cellulose acetate phthalate, hydroxypropylmethyl cellulose phthalate, hydroxymethyl cellulose acetate succinate, Eudragit (manufactured by Rohm Co., Germany, methacrylic acid / acrylic acid copolymerization) and pigments such as titanium oxide and red iron oxide are used. An intermediate layer may be provided between the enteric coating layer and the central core by a method known per se.

As the external preparation, the compound (I) of the present invention or a salt thereof can be made into a solid, semi-solid or liquid external preparation according to a method known per se. For example, as the above solid, the compound (I) or a salt thereof is used as it is, or an excipient (eg, glycol, mannitol, starch, microcrystalline cellulose, etc.), a thickener (eg, natural gums, cellulose). Derivatives, acrylic acid polymers, etc.) are added and mixed to form a powdery composition. The above-mentioned liquid form is almost the same as the case of the injection, and is an oily or aqueous suspension. If it is semi-solid,
Aqueous or oily gels or ointments are preferred. In addition, all of these are pH adjusting agents (eg, carbonic acid,
Phosphoric acid, citric acid, hydrochloric acid, sodium hydroxide, etc., preservatives (eg, paraoxybenzoic acid esters, chlorobutanol, benzalkonium chloride, etc.) and the like may be added.

For example, in the case of a suppository, the compound (I) of the present invention or a salt thereof can be made into an oily or aqueous solid, semisolid or liquid suppository according to a method known per se. Examples of the oily base used in the above composition include glycerides of higher fatty acids [eg, cacao butter, witepsols (manufactured by Dynamite Nobel), etc.], intermediate fatty acids [eg, migliols (manufactured by Dynamite Nobel)]
], Or vegetable oil (eg, sesame oil, soybean oil, cottonseed oil, etc.) and the like. Examples of the aqueous base include polyethylene glycols and propylene glycol, and examples of the aqueous gel base include natural gums, cellulose derivatives, vinyl polymers and acrylic acid polymers.

When the compound (I) or a salt thereof is used as an injection in humans, the dose may vary depending on the disease to be treated, the administration route, the age of each patient to be administered and the degree of illness. If the active ingredient is about 0.05 per day
μg to 50 mg, preferably about 0.5 μg to 10 m
g, particularly preferably about 2.5 μg to 0.25 mg is used.

[0034]

EXAMPLES The present invention will be described in more detail below with reference to Reference Examples, Examples, Experimental Examples and Preparation Examples, but the present invention is not limited thereto. The numerical value showing the mixing ratio in the mixed solvent is the volume mixing ratio of each solvent. ¹
H-NMR spectrum was measured using Varian Gemini 200 (200 MHz) using tetramethylsilane as an internal standard.
Type spectrometer, and all δ values are shown in ppm. The symbols used in this specification have the following meanings. s: singlet, d: doublet, t: triplet,
q: quartet, dd: double doublet, dt: double triplet, m: multiplet, br: a wide range of reference examples 1 Z-Gly-Gly-Gly-OH production H-Gly-Gly-Gly-OH (10.0g, 52.9 mmol, manufactured by Peptide Lab.)
Was dissolved in 4N aqueous sodium hydroxide (13.3 ml), 4N aqueous sodium hydroxide (15.9 ml) and benzyloxycarbonyl chloride (9.31 ml) were added under ice cooling, and the mixture was stirred at 20 ° C. overnight. The reaction solution was washed with ether. Under ice-cooling, the aqueous layer was adjusted to pH 3 with 5M hydrochloric acid, left to stand in a cool place overnight, and the precipitated crystals were collected by filtration, washed with cold water and dried. The obtained crystal was used without purification. Reference Example 2 Production of H-Gly-Gly-Gly-Glu (O ^t Bu) -O ^t Bu (HP-1) a) Z-Gly-Gly-Gly-OH (0.84 g, 2.6 mmol) was added to DMF (15
ml) and H-Glu (O ^t Bu) -O ^t Bu (0.68 g, 2.6 mmo
l), HOBT (386 mg, 2.9 mmol) and WSC (548 mg, 2.
(9 mmol) was added, and the mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated and suspended in a 0.2 M aqueous citric acid solution (100 ml),
It was extracted twice with ethyl acetate (100 ml). The ethyl acetate layers were combined, washed successively with 10% (W / V) aqueous ammonium chloride solution, 5% (W / V) aqueous sodium hydrogen carbonate solution, water and saturated brine, dried over anhydrous sodium sulfate, and concentrated. Ether-hexane was added to the residue and the precipitate was collected by filtration to give Z-Gly.
-Gly-Gly-Glu (O ^t Bu) -O ^t Bu (ZP-1) is white powder (0.81 g,
Yield 55%). [Α] _D ²³ + 9.6 ° (c = 0.56, in chloroform) Elemental analysis value C ₂₇ H ₄₀ N ₄ O ₉ Calculated value: C, 57.43; H, 7.14; N, 9.92 Experimental value: C, 57.60; H, 7.12; N, 9.88 b) Dissolve this powder (735 mg) in methanol (25 ml), add 10% (W / W, the same below) palladium-carbon (73 mg), and add hydrogen at normal temperature and pressure. The mixture was stirred under an air stream for 2 hours. HP-1 powder (543 mg) was obtained by removing the catalyst and distilling off the solvent.
Got [Α] _D ²³ + 13.1 ° (c = 0.51, in chloroform) Elemental analysis value Calculated as C ₁₉ H ₃₄ N ₄ O ₇ · 0.5H ₂ O: C, 51.92; H, 8.03; N, 12.75 Experimental value : C, 52.09; H, 7.89; N, 12.56

Reference Example 3 Production of H-Glu (O ^t Bu) -Gly-D-Glu (O ^t Bu) -O ^t Bu (HP-2) In the same manner as in Reference Example 2, H-Glu (O ^t Bu). ) -Gly-D-Glu (O ^t Bu) -O
^t Bu (HP-2) was manufactured. Raw material Reaction conditions Product a) 1) Fmoc-Glu (O ^t Bu) -OH WSC 1.24 g FmocP-2 (2.78 g) HOBT 876 mg (3.73 g) DCM 85 ml 2) H-Gly-D-Glu (O ^t Bu) -20 ° C 16 h O ^t Bu (1.86 g) [α] _D ²³ −12 ° (c = 0.54, in chloroform) Elemental analysis value Calculated as C ₃₉ H ₅₃ N ₃ O ₁₀ : C, 64.71; H, 7.38; N, 5.80 Experimental value: C, 64.55; H, 7.43; N, 6.09 b) This powder (2.10 g) was dissolved in DCM (63 ml), piperidine (7.0 ml) was added, and the mixture was added at 20 ° C. Stir for 1 hour. After adding ethyl acetate (300 ml), the mixture was extracted with 1N hydrochloric acid water (70 ml) and 0.03 N hydrochloric acid water (100 ml X 2). PH of water layer
After adjusting to 6.0, mix with hexane: ether (= 1: 1) (10
After washing with 0 ml X 5), adjust to pH 8.6 and wash with ethyl acetate (1
It was extracted with 50 ml X 2). The extract was dried over anhydrous sodium sulfate and concentrated to give HP-2 as a colorless oil (1.31 g). [Α] _D ²³ -17 ° (c = 0.47, in chloroform) Elemental analysis C ₂₄ H ₄₃ N ₃ O ₈ Calculated: C, 57.47; H, 8.64 ; N, 8.38 Found: C, 57.40 H, 8.73; N,
8.60

Reference Example 4 In the same manner as in Reference Example 2, H-Gly-D-Glu (O ^t Bu) -O ^t Bu (HP-
3) was manufactured. Raw material Reaction conditions Product a) 1) Z-Gly-OH WSC 569 mg ZP-3 (620 mg) HOBT 401 mg (1.23 g) DCM 23 ml 2) HD-Glu (O ^t Bu) -O ^t Bu 20 ℃ 16 h (700 mg) b) ZP-3 10% Pd-C 120 mg HP-3 (1.23 g) MeOH 40 ml (830 mg) 20 ° C 1.5 h Compound ZP-3: [α] _D ²⁰ −10.3 ° ( c = 0.52, in chloroform) elemental analysis _{C 23 H 34 N 2 O 7} · 0.25H 2 O calculated: C, 60.71; H, 7.64 ; N, 6.16 Found: C, 60.79; H, 7.68 ; N , 6.28 Compound HP-3: [α] _D ²⁰ -17.9 ° (c = 0.52, in chloroform) Elemental analysis value Calculated as C ₁₅ H ₂₈ N ₂ O ₅ : C, 56.94; H, 8.92; N, 8.85 experiment Values: C, 56.81; H, 8.95; N, 9.04

Reference Example 5 In the same manner as in Reference Example 2, H-Gly-Glu (O ^t Bu) -Glu (O ^t Bu) -O ^t.
Bu (HP-4) was manufactured. Raw material Reaction conditions Product a) 1) Z-Gly-OH WSC 318 mg ZP-4 (347 mg) HOBT 224 mg (930 mg) DCM 22 ml 2) H-Glu (O ^t Bu) -Glu (O ^t Bu ) -O ^t Bu 20 ° C 16 h (670 mg) b) ZP-4 10% Pd-C 90 mg HP-4 (870 mg) MeOH 30 ml (650 mg) 20 ° C 2 h Compound ZP-4: melting point 92.4 -92.7 ℃ [α] _D ²⁴ −3.5 ° (c = 0.51, in chloroform) Elemental analysis value Calculated as C ₃₂ H ₄₉ N ₃ O ₁₀・ 0.25H ₂ O: C, 60.03; H, 7.79; N, 6.56 Experimental value: C, 60.18; H, 7.81; N, 6.26 Compound HP-4: [α] _D ²⁴ −6.8 ° (c = 0.54, in chloroform) Elemental analysis value C ₂₄ H ₄₃ N ₃ O ₈ 0.25H ₂ O Calculated: C, 56.95; H, 8.66; N, 8.30 Experimental: C, 56.92; H, 8.74; N, 8.06

Reference Example 6 In the same manner as in Reference Example 2, H-Glu (O ^t Bu) -Gly-Glu (O ^t Bu) -O ^t.
Bu (HP-5) was manufactured. Raw material Reaction conditions Product a) 1) Z-Glu (O ^t Bu) -OH WSC 447 mg ZP-5 (785 mg) HOBT 315 mg (1.20 g) DCM 22 ml 2) H-Gly-Glu (O ^t Bu ) -O ^t Bu 20 ° C 16 h (670 mg) b) ZP-5 10% Pd-C 110 mg HP-5 (1.10 g) MeOH 40 ml (840 mg) 20 ° C 2 h Compound ZP-5: (α ] _D ²⁴ + 4.2 ° (c = 0.53, in chloroform) Elemental analysis value C ₃₂ H ₄₉ N ₃ O ₁₀ Calculated value: C, 60.46; H, 7.77;
N, 6.61 Experimental: C, 60.48; H, 7.78;
N, 6.88 Compound HP-5: [α] _D ²⁴ + 7.0 ° (c = 0.52, in chloroform) Elemental analysis value C ₂₄ H ₄₃ N ₃ O ₈ 0.25H ₂ O Calculated value: C, 56.95 H, 8.66; N, 8.30 Experimental value: C, 56.94; H, 8.60; N, 8.04

Reference Example 7 In the same manner as in Reference Example 2, H-Gly-Lys (Boc) -Gly-O ^t Bu (HP-
6) was synthesized. Raw material Reaction conditions Product a) 1) Z-Gly-OH WSC 745 mg ZP-6 (813 mg) HOBT 525 mg (1.90 g) DMF 20 ml 2) H-Lys (Boc) -Gly-O ^t Bu 20 ° C 13 h (1.27 g) b) ZP-6 10% Pd / C 154 mg HP-6 (1.54 g) MeOH 40 ml (1.15 g) 20 ° C 5 h Compound ZP-6: [α] _D ²⁴ -13.4 ° ( c = 0.58, in chloroform) Elemental analysis value Calculated as C ₂₇ H ₄₂ N ₄ O ₈ 0.5H ₂ O: C, 57.95; H, 7.74; N, 10.01 Experimental value: C, 57.73; H, 7.44; N , 10.03 Compound HP-6: [α] _D ²⁴ −23.8 ° (c = 0.58, in chloroform) Elemental analysis value Calculated as C ₁₉ H ₃₆ N ₄ O ₆ · H ₂ O: C, 52.52; H, 8.81; N, 12.89 Experimental value: C, 52.72; H, 8.81; N, 12.86

Reference Example 8 (2R, 6R) -2-Fmoc Amino-6,7-bis (PamO) -4-THT-OH (Fmoc
Synthesis of GC-1) a) Literature [JW Metzge (JW Metzge
r) et al., International Journal of Peptide and Protein Research (Int. J. Peptide P
rotein Res.), Vol. 38, 545 pp., was prepared by the method described in 1991] (Fmoc- (R) -Cys-O t Bu) 2 (10.0 g, 12.5 m
mol) in DCM (80 ml), and under ice cooling, zinc powder (3.27
g, 50.0 mmol) and MeOH-36% (W / V) hydrochloric acid-concentrated sulfuric acid (10
0: 7: 1) mixture (hereinafter referred to as acid mixture) (40 ml)
Was added and the mixture was stirred at 20 ° C. for 20 minutes. Into the reaction mixture
Add (R)-(+)-glycidol (8.29 ml, 125 mmol) and add 4
The mixture was stirred at 0 ° C for 2.5 hours. The reaction mixture was concentrated to 50 ml, and the insoluble material was filtered off. Saturated saline solution in the filtrate
(200 ml) was added and extracted with DCM (2 x 300 ml), DC
The M layers were combined, dried over anhydrous sodium sulfate, and concentrated. The residue was subjected to silica gel column chromatography, and eluted successively with ethyl acetate-hexane (1: 1, 3: 1). Fractions containing the desired product were collected and concentrated, and (2R, 6R) -2-Fmoc
Amino-6,7-dihydroxy-4-THT-O ^t Bu (FmocGC-1a) was obtained as a white powder (10.9 g, yield 92%). (Α) _D ²¹ −8.8 ° (c = 0.65, in chloroform) Elemental analysis value Calculated as C ₂₅ H ₃₁ NO ₆ S ・ 0.5H ₂ O: C, 62.22; H, 6.68; N, 2.90; S, 6.64 Experimental value: C, 62.14; H, 6.66; N, 2.81; S, 6.54 b) Compound obtained above (FmocGC-1a) (3.90 g, 8.23
mmol) in THF (70 ml) and palmitic acid (6.33
g, 24.7 mmol), DIC (3.87 ml, 24.7 mmol) and 4-
Dimethylaminopyridine (DMAP, 402 mg, 3.29 mmol)
Was added, and the mixture was stirred at 20 ° C. for 13 hours. The reaction mixture was concentrated, the residue was suspended in 10% (W / V) aqueous citric acid (200 ml), and extracted with ethyl acetate (400 ml). The ethyl acetate layer was washed with water and concentrated, and the residue obtained was subjected to silica gel column chromatography, ethyl acetate-hexane (1: 2
It was eluted sequentially at 0, 1:10). Fractions containing the desired product were collected, concentrated, and crystallized from hexane to give (2R, 6R) -2-Fmocamino-6,7-bis (PamO) -4-THT-O ^t Bu (FmocGC-1b). Was obtained as colorless crystals (4.76 g, yield 61%). Melting point 58.0 ° C Elemental analysis value Calculated as C ₅₇ H ₉₁ NO ₈ S: C, 72.03; H, 9.65; N, 1.47; S, 3.37 Experimental value: C, 71.94; H, 9.58; N, 1.43 S, 3.36 c) Compound obtained above (FmocGC-1b) (2.50 g, 2.63
mmol) was dissolved in TFA (50 ml) and the mixture was left standing at 20 ° C for 1.5 hours. The reaction mixture was concentrated and the residue was washed with ethyl acetate (500
ml). The solution was washed with water, dried over anhydrous sodium sulfate, and concentrated. The obtained crystals were recrystallized from ethyl acetate to give FmocGC-1 as colorless crystals (2.20 g, yield 94%). Melting point 90.0 ° C [α] _D ²⁰ + 12.9 ° (c = 0.73, in chloroform) Elemental analysis value Calculated as C ₅₃ H ₈₃ NO ₈ S: C, 71.18; H, 9.35; N, 1.57; S, 3.59 experiment Values: C, 71.23; H, 9.12; N, 1.54; S, 3.47

Reference Example 9 (2R, 6R) -2-Amino-6,7-bis (PamO) -4-THT-Gly-Gly-Gly-
Synthesis of Glu-OH (HGCP-1) a) Compound (FmocGC-1) (1.31 g) synthesized in Reference Example 8
Dissolve in DMF (19 ml), and under ice cooling HONB (288 mg), DIC (2
50 μl) and the H-Gly-Gly-Gly-Glu (O ^t B obtained in Reference Example 2).
u) -O ^t Bu (HP-1) (630 mg) was added, and the mixture was stirred at 20 ° C. for 15 hr. After the reaction mixture was concentrated, the residue was dissolved in ethyl acetate and washed successively with 0.2 M aqueous citric acid solution, 10% aqueous ammonium chloride solution, 5% aqueous sodium hydrogen carbonate solution, water and saturated brine. The organic layer was dried over anhydrous sodium sulfate and concentrated. Acetonitrile was added to the residue and collected by filtration,
(2R, 6R) -2-Fmoc amino-6,7-bis (PamO) -4-THT-Gly-Gly-
Gly-Glu (O ^t Bu) -O ^t Bu (FmocGCP-1) white powder (1.64
g, yield 86%) was obtained. (Α) _D ²³ −6.4 ° (c = 0.50, in chloroform) Elemental analysis C ₇₂ H ₁₁₅ N ₅ O ₁₄ S Calculated value: C, 66.18; H, 8.87; N, 5.36; S, 2.45 Experimental value: C, 66.03; H, 8.87; N, 5.31; S, 2.22 b) The compound (FmocGCP-1) (1.18 g) obtained above was added to D
Dissolve in CM (12 ml), add piperidine (1.2 ml),
The mixture was stirred at 0 ° C for 1.3 hours. The reaction mixture was concentrated, subjected to silica gel chromatography, and eluted successively with chloroform-methanol (49: 1, 19: 1, 14: 1). Fractions containing the desired product were collected and concentrated, and (2R, 6R) -2-amino-6,7-bis (PamO) -4-THT-Gly-Gly-Gly-Glu (O ^t Bu) -O ^t Bu (HGCP-1)
Of white powder (0.92 g, yield 94%) was obtained. (Α) _D ²³ −6.7 ° (c = 0.63, in chloroform) Elemental analysis value Calculated as C ₅₇ H ₁₀₅ N ₅ O ₁₂ S ・ H ₂ O: C, 62.09; H, 9.78; N, 6.35; S, 2.91 Experimental value: C, 62.12; H, 9.59; N, 6.36; S, 2.87

Reference Example 10 In the same manner as in Reference Example 9, (2R, 6R) -2-Fmocamino-6,7-bis (PamO) -4-THT-Glu (O ^t Bu) -Gly-D-Glu. (O ^t Bu) -O ^t Bu (Fmo
cGCP-2), (2R, 6R) -2-Amino-6,7-bis (PamO) -4-THT-Glu
(O ^t Bu) -Gly-D-Glu (O ^t Bu) -O ^t Bu (HGCP-2) was synthesized. Raw material (g) Reaction conditions Product (g) a) FmocGC-1 HP-2 1.31 g FmocGCP-2 (2.04) HONB 453 mg (2.72) DIC 0.40 ml DMF 20 ml 20 ° C 16 hb) FmocGCP-2 Piperidine 2.0 ml HGCP-2 (2.00) DCM 18 ml (1.75) 20 ℃ 1 h Silica gel (ethyl acetate-methanol) 10: 0, 9: 1 compound Fmoc GCP-2: [α] _D ²³ −7.0 ° (c = 0.56, in chloroform ) Elemental analysis value Calculated as C ₇₇ H ₁₂₄ N ₄ O ₁₅ S: C, 67.12; H, 9.07; N, 4.07; S, 2.33 Experimental value: C, 67.01; H, 9.19; N, 4.01; S, 2.31 Compound HGCP-2: [α] _D ²³ −18 ° (c = 0.53, in chloroform) Elemental analysis value Calculated as C ₆₂ H ₁₁₄ N ₄ O ₁₃ S: C, 64.44; H, 9.94; N, 4.85; S , 2.77 Experimental value: C, 64.23; H, 9.95; N, 4.94; S, 2.64

Reference Example 11 In the same manner as in Reference Example 9, (2R, 6R) -2-Fmocamino-6,7-bis (PamO) -4-THT-Gly-D-Glu (O ^t Bu) -O ^t Bu (FmocGCP-3),
(2R, 6R) -2-Amino-6,7-bis (PamO) -4-THT-Gly-D-Glu (O ^t
Bu) -O ^t Bu (HGCP-3) was synthesized. Raw material (g) Reaction conditions Product (g) a) FmocGC-1 HP-3 830 mg FmocGCP-3 (2.12) HONB 471 mg (2.18) DIC 412 μl DMF 21 ml 20 ℃ 16 hb) FmocGCP-3 Piperidine 2.0 ml HGCP -3 (2.00) DCM 18 ml (1.72) 20 ℃ 1 h Silica gel (chloroform-methanol) 97: 3 Compound FmocGCP-3: [α] _D ²⁴ -12.5 ° (c = 0.55, in chloroform) Elemental analysis C ₆₈ Calculated as H ₁₀₉ N ₃ O ₁₂ S: C, 68.48; H, 9.21; N, 3.52; S, 2.69 Experimental value: C, 68.31; H, 9.25; N, 3.72; S, 2.48 Compound HGCP-3: ( α] _D ²⁴ −16.4 ° (c = 0.53, in chloroform) Elemental analysis value C ₅₃ H ₉₉ N ₃ O ₁₀ S ・ 0.8H ₂ O Calculated value: C, 64.64; H, 10.30; N, 4.27; S, 3.26 Experimental value: C, 64.62; H, 10.25;
N, 4.08; S, 3.22.

Reference Example 12 In the same manner as in Reference Example 9, (2R, 6R) -2-Fmocamino-6,7-bis (PamO) -4-THT-Gly-Glu (O ^t Bu) -Glu (O ^t Bu) -O ^t Bu (FmocG
CP-4), (2R, 6R) -2-Amino-6,7-bis (PamO) -4-THT-Gly-G
lu (O ^t Bu) -Glu (O ^t Bu) -O ^t Bu (HGCP-4) was synthesized. Raw material (g) Reaction conditions Product (g) a) FmocGC-1 HP-4 590 mg FmocGCP-4 (0.95) HONB 210 mg (1.22) DIC 185 μl DMF 9.5 ml 20 ° C 16 hb) FmocGCP-4 Piperidine 1.1 ml HGCP -4 (1.10) DCM 9.9 ml (0.97) 20 ℃ 1 h Silica gel (chloroform-methanol) 97: 3 Compound FmocGCP-4: [α] _D ²⁴ −7.0 ° (c = 0.50, in chloroform) Elemental analysis value C ₇₇ Calculated as H ₁₂₄ N ₄ O ₁₅ S: C, 67.12; H, 9.07; N, 4.07; S, 2.33 Experimental value: C, 67.16; H, 9.05; N, 4.10; S, 2.40 Compound HGCP-4: ( α] _D ²⁴ −10.6 ° (c = 0.50, in chloroform) Elemental analysis value Calculated as C ₆₂ H ₁₁₄ N ₄ O ₁₃ S ・ 0.5H ₂ O: C, 63.94; H, 9.95; N, 4.81; S, 2.75 Experimental value: C, 63.91; H, 9.80; N, 4.74; S, 2.71

Reference Example 13 In the same manner as in Reference Example 9, (2R, 6R) -2-Fmocamino-6,7-bis (PamO) -4-THT-Glu (O ^t Bu) -Gly-Glu (O ^t Bu) -O ^t Bu (FmocG
CP-5), (2R, 6R) -2-Amino-6,7-bis (PamO) -4-THT-Glu (O
^t Bu) -Gly-Glu (O ^t Bu) -O ^t Bu (HGCP-5) was synthesized. Raw material (g) Reaction conditions Product (g) a) FmocGC-1 HP-5 770 mg FmocGCP-5 (1.24) HONB 274 mg (1.63) DIC 241 μl DMF 12.0 ml 20 ° C 16 hb) FmocGCP-5 piperidine 1.5 ml HGCP -5 (1.50) DCM 13.5 ml (1.34) 20 ℃ 1 h Silica gel (chloroform-methanol) 97: 3 Compound FmocGCP-5: [α] _D ²⁴ -1.7 ° (c = 0.51, in chloroform) Elemental analysis C ₇₇ Calculated as H ₁₂₄ N ₄ O ₁₅ S: C, 67.12; H, 9.07; N, 4.07; S, 2.33 Experimental value: C, 66.97; H, 9.12; N, 4.06; S, 2.25 Compound HGCP-5: ( α] _D ²⁴ −9.6 ° (c = 0.53, in chloroform) Elemental analysis value Calculated as C ₆₂ H ₁₁₄ N ₄ O ₁₃ S ・ 1H ₂ O: C, 63.45; H, 9.96; N, 4.77; S, 2.73 Experimental value: C, 63.31; H, 9.81; N, 4.82; S, 2.65

Reference Example 14 In the same manner as in Reference Example 9, (2R, 6R) -2-Fmocamino-6,7-bis (PamO) -4-THT-Gly-Lys (Boc) -Gly-O ^t Bu (FmocGCP-6),
(2R, 6R) -2-Amino-6,7-bis (PamO) -4-THT-Gly-Lys (Boc)
-Gly-O ^t was Bu the (HGCP-6) was synthesized. Raw material (g) Reaction conditions Product (g) a) FmocGC-1 HP-6 1.01 g FmocGCP-6 (1.97) HONB 435 mg (2.24) DIC 0.38 ml DMF 30 ml 20 ℃ 13 hb) FmocGCP-6 piperidine 2.0 ml HGCP-6 (2.00) DCM 20 ml (1.23) 20 ° C 1.5 h Silica gel (chloroform-methanol) 50: 1 compound Fmoc GCP-6: [α] _D ²⁵ -10.0 ° (c = 0.66 in chloroform) Elemental analysis value C Calculated as ₇₂ H ₁₁₇ N ₅ O ₁₃ S: C, 66.89; H, 9.12; N, 5.42; S, 2.48 Experimental value: C, 67.10; H, 9.37; N, 5.18; S, 2.49 Compound HGCP-6: [Α] _D ²⁵ −15.2 ° (c = 0.56, in chloroform) Elemental analysis value Calculated as C ₅₇ H ₁₀₇ N ₅ O ₁₁ S: C, 63.95; H, 10.07; N, 6.54; S, 3.00 Experimental value: C, 63.70; H, 10.20; N, 6.50; S, 2.99

Example 1 (2R, 6R) -2- (H-Gly-amino) -6,7-bis (PamO) -4-THT-Glu-Gly-
Synthesis of D-Glu-OH (Compound 1) a) Compounds HGCP-2 (900 mg) and Boc-Gly-OH (150 mg)
Was dissolved in DCM (10 ml) and HOBT (116 mg) and W were added under ice-cooling.
SC (165 mg) was added, and the mixture was stirred at 20 ° C for 20 hr. The reaction mixture was concentrated, and the residue was dissolved in ethyl acetate (100 ml). Add this solution to 10% (W / V) aqueous ammonium chloride, 2% (W / V)
/ V) After washing successively with aqueous sodium hydrogen carbonate and water, the ethyl acetate layer was dried over anhydrous sodium sulfate and concentrated. The residue was suspended in acetonitrile and collected by filtration, (2R, 6R) -2- (Boc-Gly-amino) -6,7-bis (PamO) -4-THT-Glu (O ^t Bu) -Gly- D-Glu (O ^t Bu) -O
A white powder of ^t Bu (compound 1a) (811 mg, yield 80%) was obtained. (Α) _D ²⁵ −8.5 ° (c = 0.59, in chloroform) Elemental analysis value Calculated as C ₆₉ H ₁₂₅ N ₅ O ₁₆ S: C, 63.13; H, 9.60; N, 5.33; S, 2.44 Experimental value: C, 63.04; H, 9.52; N, 5.26; S, 2.14 b) Compound 1a (715 mg) was dissolved in TFA (8 ml), 20
It was allowed to stand at ℃ for 2 hours. The reaction mixture was concentrated, the residue was suspended in acetonitrile and collected by filtration to give (2R, 6R) -2- (H
A white powder (580 mg, yield 98%) of -Gly-amino) -6,7-bis (PamO) -4-THT-Glu-Gly-D-Glu-OH (Compound 1) was obtained. (Α) _D ²⁵ -17.7 ° (c = 0.69, 5% TFA-in chloroform) Elemental analysis value Calculated as C ₅₂ H ₉₃ N ₅ O ₁₄ S ・ 2H ₂ O: C, 57.81; H, 9.05; N, 6.48; S, 2.97 Experimental value: C, 57.66; H, 9.22; N, 6.23; S, 2.72

Example 2 (2R, 6R) -2- (H-Glu-amino) -6,7-bis (PamO) -4-THT-Glu-Gly-
Synthesis of D-Glu-OH (Compound 2) a) Compounds HGCP-2 (2.50 g) and Fmoc-Glu (O ^t Bu) -OH (1.
06 g) was dissolved in DCM (25 ml) and HOBT (322 mg) under ice cooling.
And WSC (456 mg) were added, and the mixture was stirred at 20 ° C for 16 hr. The reaction mixture was concentrated and the residue was ethyl acetate (200 ml)
Dissolved in. This solution was washed successively with 10% (W / V) ammonium chloride aqueous solution, 2% (W / V) aqueous sodium hydrogen carbonate solution and water, and then the ethyl acetate layer was dried over anhydrous sodium sulfate and concentrated. The residue was suspended in acetonitrile and collected by filtration, (2R, 6R) -2- (Fmo
c-Glu (O ^t Bu) -amino) -6,7-bis (PamO) -4-THT-Glu (O ^t Bu) -Gly
-D-Glu (O ^t Bu) -O ^t Bu (Compound 2a) white powder (3.00 g,
Yield 89%) was obtained. (Α) _D ²⁵ −9.7 ° (c = 0.74, in chloroform) Elemental analysis value Calculated as C ₈₆ H ₁₃₉ N ₅ O ₁₈ S ・ H ₂ O: C, 65.33; H, 8.99; N, 4.43; S, 2.03 Experimental value: C, 65.58; H, 8.96; N, 4.49; S, 2.08 b) Compound 2a (2.90 g) was dissolved in DCM (20 ml), piperidine (2.9 ml) was added, and the mixture was stirred at 20 ° C for 1.5 hours. did. The reaction mixture was concentrated, and then subjected to silica gel column chromatography, chloroform-methanol (50: 1,20:
Elution was carried out in sequence 1). Fractions containing the desired product were collected and concentrated to give (2R, 6R) -2- (H-Glu (O ^t Bu) -amino) -6,7-bis (PamO) -4-TH.
A white powder (1.88 g, yield 76%) of T-Glu (O ^t Bu) -Gly-D-Glu (O ^t Bu) -O ^t Bu (Compound 2b) was obtained. (Α) _D ²⁵ −13.1 ° (c = 0.56, in chloroform) Elemental analysis value Calculated as C ₇₁ H ₁₂₉ N ₅ O ₁₆ S ・ 0.5H ₂ O: C, 63.17; H, 9.71; N, 5.19; S , 2.38 Experimental value: C, 63.25; H, 9.66; N, 4.95; S, 2.24 c) Compound 2b (760 mg) was dissolved in TFA (8 ml), 20
It was allowed to stand at ℃ for 2 hours. The reaction mixture was concentrated, the residue was suspended in acetonitrile and collected by filtration to give (2R, 6R) -2- (H
A white powder (612 mg, yield 97%) of -Glu-amino) -6,7-bis (PamO) -4-THT-Glu-Gly-D-Glu-OH (Compound 2) was obtained. (Α) _D ²⁵ −4.0 ° (c = 0.56, in 5% TFA-chloroform) Elemental analysis value Calculated as C ₅₅ H ₉₇ N ₅ O ₁₆ S ・ 2H ₂ O: C, 57.32; H, 8.83; N, 6.08; S, 2.78 Experimental value: C, 57.44; H, 8.24; N, 5.84; S, 2.81

Example 3 (2R, 6R) -2- (H-Gly-Glu-amino) -6,7-bis (PamO) -4-THT-
Synthesis of Glu-Gly-D-Glu-OH (Compound 3) Compound (2R, 6R) -2-
(Boc-Gly-Glu-amino) -6,7-bis (PamO) -4-THT-Glu (O ^t B
u) -Gly-D-Glu (O ^t Bu) -O ^t Bu (Compound 3a) and (2R, 6R)-
2- (H-Gly-Glu-amino) -6,7-bis (PamO) -4-THT-Glu-Gly-
D-Glu-OH (Compound 3) was synthesized. Raw material (g) Reaction conditions Product (g) a) Boc-Gly-OH 2b 900 mg 3a (0.13) HOBT 99 mg (0.96) WSC 142 mg DCM 10 ml 20 ° C 14 hb) 3a TFA 8.0 ml 3 (0.73) 20 ° C 2 h (0.59) Compound 3a: [α] _D ²⁵ -15.3 ° (c = 0.57, in chloroform) Elemental analysis C ₇₈ H ₁₄₀ N ₆ O ₁₉ S ・ Calculated as 0.5H ₂ O: C, 62.16 H, 9.43; N, 5.58; S, 2.13 Experimental value: C, 62.24; H, 9.15; N, 5.31; S, 2.06 Compound 3: [α] _D ²⁵ -27.8 ° (c = 0.59, 5% TFA- Chloroform) Elemental analysis value Calculated as C ₅₇ H ₁₀₆ N ₆ O ₂₀ S ・ 0.5H ₂ O: C, 55.37; H, 8.72; N, 6.80; S, 2.59 Experimental value: C, 55.28; H, 8.35; N, 6.46; S, 2.39

Example 4 (2R, 6R) -2- (H-Glu-Gly-amino) -6,7-bis (PamO) -4-THT-
Synthesis of Glu-Gly-D-Glu-OH (Compound 4) In the same manner as in Example 2, (2R, 6R) -2- (Fmoc-Gly-amino) -6,7-
Bis (PamO) -4-THT-Glu (O ^t Bu) -Gly-D-Glu (O ^t Bu) -O ^t Bu
(Compound 4a), (2R, 6R) -2- (H-Gly-amino) -6,7-bis (Pam
O) -4-THT-Glu (O ^t Bu) -Gly-D-Glu (O ^t Bu) -O ^t Bu (Compound 4
b) was synthesized. Further, using compound 4b as a raw material, Example 2
Similar to (2R, 6R) -2- (Fmoc-Glu (O ^t Bu) -Gly-amino) -6,7
-Bis (PamO) -4-THT-Glu (O ^t Bu) -Gly-D-Glu (O ^t Bu) -O ^t Bu
(Compound 4c), (2R, 6R) -2- (H-Glu (O ^t Bu) -Gly-amino) -6,7
-Bis (PamO) -4-THT-Glu (O ^t Bu) -Gly-D-Glu (O ^t Bu) -O ^t Bu
(Compound 4d) and (2R, 6R) -2- (H-Glu-Gly-amino) -6,
7-bis (PamO) -4-THT-Glu-Gly-D-Glu-OH (Compound 4) was synthesized. Raw material (g) Reaction conditions Product (g) a) Fmoc-Gly-OH HGCP-2 2.00 g 4a (0.57) HOBT 257 mg (2.08) WSC 365 mg DCM 20 ml 20 ° C 16 hb) 4a Piperidine 2.0 ml 4b ( 2.03) DCM 20 ml (1.63) 20 ° C 1.5 h Silica gel (chloroform-methanol) 50: 1 to 20: 1 c) Fmoc-Glu (O ^t Bu)-4b 0.75 g 4c OH HOBT 92 mg (0.86) (0.30) WSC 130 mg DCM 8.0 ml 20 ℃ 16 hd) 4c Piperidine 0.8 ml 4d (0.81) DCM 8.0 ml (0.68) 20 ℃ 1.5 h Silica gel (chloroform-methanol) 50: 1 to 10: 1 e) 4d TFA 6.0 ml 4 ( 0.60) 20 ° C 1.5 h (0.49) Compound 4a: [α] _D ²⁴ -9.9 ° (c = 0.53, in chloroform) Elemental analysis value Calculated as C ₇₉ H ₁₂₇ N ₅ O ₁₆ S: C, 66.13; H, 8.92; N, 4.88; S, 2.23 Experimental value: C, 66.01; H, 9.01; N, 4.64; S, 2.26 Compound 4b: (α) _D ²⁴ -11.2 ° (c = 0.57, in chloroform) Elemental analysis value C Calculated for ₆₄ H ₁₁₇ N ₅ O ₁₄ S: C, 63.39; H, 9.72;
N, 5.78; S, 2.64 Experimental value: C, 63.17; H, 10.02;
N, 5.51; S, 2.62 Compound 4c: [α] _D ²⁴ -10.6 ° (c = 0.54, in chloroform) Elemental analysis value Calculated as C ₈₈ H ₁₄₂ N ₆ O ₁₉ S: C, 65.24; H, 8.83; N, 5.19; S, 1.98 Experimental value: C, 65.12; H, 8.90; N, 5.03; S, 2.03 Compound 4d: (α) _D ²⁴ -11.7 ° (c = 0.52, in chloroform) Elemental analysis Value calculated as C ₇₃ H ₁₃₂ N ₆ O ₁₇ S: C, 62.72; H, 9.52; N, 6.01; S, 2.29 Experimental value: C, 62.62; H, 9.70; N, 5.87; S, 2.31 Compound 4: (Α) _D ²⁴ -9.1 ° (c = 0.59, 5% TFA-in chloroform) Elemental analysis value Calculated as C ₅₇ H ₁₀₀ N ₆ O ₁₇ S ・ 3H ₂ O: C, 55.77; H, 8.70; N, 6.85; S, 2.61 Experimental value: C, 55.63; H, 8.61; N, 6.46; S, 2.78

Example 5 (2R, 6R) -2- (H-Tyr-amino) -6,7-bis (PamO) -4-THT-Glu-
Synthesis of Gly-D-Glu-OH (Compound 5) In the same manner as in Example 2, (2R, 6R) -2- (Fmoc-Tyr ( ^t Bu) -amino)
-6,7-Bis (PamO) -4-THT-Glu (O ^t Bu) -Gly-D-Glu (O ^t Bu) -O ^t
Bu (compound 5a), (2R, 6R) -2- (H-Tyr ( ^t Bu) -amino) -6,7-
Bis (PamO) -4-THT-Glu (O ^t Bu) -Gly-D-Glu (O ^t Bu) -O ^t Bu
(Compound 5b) and (2R, 6R) -2- (H-Tyr-amino) -6,7-
Bis (PamO) -4-THT-Glu-Gly-D-Glu-OH (Compound 5) was synthesized. Raw material (g) Reaction conditions Product (g) a) Fmoc-Tyr ( ^t Bu)-HGCP-2 1.2 g 5a OH HOBT 154 mg (1.49) (0.53) WSC 219 mg DCM 20 ml 20 ℃ 21 hb) 5a Piperidine 1.4 ml 5b (1.44) DCM 14 ml (1,24) 20 ° C 1.5 h Silica gel (chloroform-methanol) 50: 1 to 10: 1 c) 5b TFA 11.7 ml 5 (1.17) 20 ° C 2 h (0.98) Compound 5a : [Α] _D ²⁵ -11.6 ° (c = 0.59, in chloroform) Elemental analysis C ₉₀ H ₁₄₁ N ₅ O ₁₇ S ・ 0.5H ₂ O Calculated: C, 67.30; H, 8.91; N, 4.36; S, 2.00 Experimental value: C, 67.32; H, 8.74; N, 4.20; S, 2.03 Compound 5b: [α] _D ²⁵ -25.0 ° (c = 0.59, in chloroform) Elemental analysis value C ₇₅ H ₁₃₁ N ₅ O Calculated as ₁₅ S ・ 0.5H ₂ O: C, 65.09; H, 9.61; N, 5.06; S, 2.32 Experimental value: C, 65.25; H, 9.68; N, 5.07; S, 2.27 Compound 5: (α) _D ²⁵ + 7.0 ° (c = 0.61, in 5% TFA-chloroform) Elemental analysis value Calculated as C ₅₉ H ₉₉ N ₅ O ₁₅ S ・ 2.5H ₂ O: C, 59.27; H, 8.77; N, 5.86; S, 2.68 experimental value: C, 59.45; H, 8.48; N, 5.60; S, 2.54

Example 6 (2R, 6R) -2- (H-Lys-amino) -6,7-bis (PamO) -4-THT-Glu-
Synthesis of Gly-D-Glu-OH (Compound 6) In the same manner as in Example 2, (2R, 6R) -2- (Fmoc-Lys (Boc) -amino)
-6,7-Bis (PamO) -4-THT-Glu (O ^t Bu) -Gly-D-Glu (O ^t Bu) -O ^t
Bu (6a), (2R, 6R) -2- (H-Lys (Boc) -amino) -6,7-bis (Pam
O) -4-THT-Glu (O ^t Bu) -Gly-D-Glu (O ^t Bu) -O ^t Bu (6b) and (2R, 6R) -2- (H-Lys-amino) -6, 7-bis (PamO) -4-THT-
Glu-Gly-D-Glu-OH (Compound 6) was synthesized. Raw material (g) Reaction conditions Product (g) a) Fmoc-Lys (Boc)-HGCP-2 1.2 g 6a OH HOBT 154 mg (1.47) (0.53) WSC 219 mg DCM 15 ml 20 ° C 22 hb) 6a Piperidine 1.4 ml 6b (1.42) DCM 14 ml (1.13) 20 ° C 1.5 h Silica gel (chloroform-methanol) 50: 1 to 10: 1 c) 6b TFA 10 ml 6 (1.05) 20 ° C 2 h (0.77) Compound 6a: (α ] _D ²⁴ -9.7 ° (c = 0.52, in chloroform) Elemental analysis value Calculated as C ₈₈ H ₁₄₄ N ₆ O ₁₈ S: C, 65.81; H, 9.04; N, 5.23 Experimental value: C, 65.60; H, 8.87; N, 5.20 Compound 6b: (α) _D ²⁴ -13.4 ° (c = 0.69, in chloroform) Elemental analysis C ₇₃ H ₁₃₄ N ₆ O ₁₆ S Calculated: C, 63.36; H, 9.76; N, 6.07; S, 2.32 Experimental value: C, 63.22; H, 9.89; N, 5.90; S, 2.31 Compound 6: [α] _D ²⁴ + 0.9 ° (c = 0.80, 5% TFA-in chloroform) Elemental analysis value C Calculated as ₅₆ H ₁₀₂ N ₆ O ₁₄ S ・ 1.5H ₂ O: C, 58.87; H, 9.26; N, 7.36; S, 2.81 Experimental value: C, 58.91; H, 9.53; N, 7.39; S, 2.76

Example 7 (2R, 6R) -2- (H-β-Ala-amino) -6,7-bis (PamO) -4-THT-G
Synthesis of lu-Gly-D-Glu-OH (Compound 7) In the same manner as in Example 1, (2R, 6R) -2- (Boc-β-Ala-amino) -6,
7-bis (PamO) -4-THT-Glu (O ^t Bu) -Gly-D-Glu (O ^t Bu) -O ^t Bu
(Compound 7a) and (2R, 6R) -2- (H-β-Ala-amino) -6,7
-Bis (PamO) -4-THT-Glu-Gly-D-Glu-OH (Compound 7) was synthesized. Raw material (g) Reaction conditions Product (g) a) Boc-β-Ala-OH HGCP-2 800 mg 7a (0.14) HOBT 103 mg (0.80) WSC 146 mg DCM 8.0 ml 20 ° C 15 hb) 7a TFA 7.5 ml 7 (0.75) 20 ℃ 2 h (0.59) Compound 7a: [α] _D ²⁴ -8.2 ° (c = 0.57, in chloroform) Elemental analysis value Calculated as C ₇₀ H ₁₂₇ N ₅ O ₁₆ S: C, 63.37; H, 9.65; N, 5.28; S, 2.42 Experimental value: C, 63.37; H, 9.43; N, 5.17; S, 2.45 Compound 7: (α) _D ²⁴ -14.3 ° (c = 0.60, 5% TFA-chloroform (Middle) Elemental analysis value Calculated as C ₅₃ H ₉₅ N ₅ O ₁₄ S ・ 2H ₂ O: C, 58.16; H, 9.12; N, 6.40; S, 2.93 Experimental value: C, 58.17; H, 9.06; N, 6.56; S, 2.84

Example 8 (2R, 6R) -2- (H-Gly-amino) -6,7-bis (PamO) -4-THT-Glu-
Synthesis of Gly-Glu-OH (Compound 8) In the same manner as in Example 1, (2R, 6R) -2- (Boc-Gly-amino) -6,7-
Bis (PamO) -4-THT-Glu (O ^t Bu) -Gly-Glu (O ^t Bu) -O ^t Bu (Compound 8a) and (2R, 6R) -2- (H-Gly-amino) -6 , 7-bis
(PamO) -4-THT-Glu-Gly-Glu-OH (Compound 8) was synthesized. Raw material (g) Reaction conditions Product (g) a) Boc-Gly-OH HGCP-5 900 mg 8a (0.15) HOBT 116 mg (0.74) WSC 165 mg DCM 10 ml 20 ° C 16 hb) 8a TFA 6.5 ml 8 ( 0.65) 20 ℃ 2 h (0.53) Compound 8a: [α] _D ²⁴ -9.3 ° (c = 0.61, in chloroform) Elemental analysis value Calculated as C ₆₉ H ₁₂₅ N ₅ O ₁₆ S: C, 63.13; H, 9.60; N, 5.34; S, 2.44 Experimental value: C, 63.08; H, 9.31; N, 5.20; S, 2.48 Compound 8: (α) _D ²⁴ -15.1 ° (c = 0.60, 5% TFA-in chloroform) Elemental analysis value Calculated as C ₅₂ H ₉₃ N ₅ O ₁₄ S ・ 1.5H2O: C, 58.29; H, 9.03; N, 6.54; S, 2.99 Experimental value: C, 58.25; H, 8.88; N, 6.42; S , 2.82

Example 9 (2R, 6R) -2- (H-Gly-amino) -6,7-bis (PamO) -4-THT-Gly-
Synthesis of Glu-Glu-OH (Compound 9) In the same manner as in Example 1, (2R, 6R) -2- (Boc-Gly-amino) -6,7-
Bis (PamO) -4-THT-Gly-Glu (O ^t Bu) -Glu (O ^t Bu) -O ^t Bu (Compound 9a) and (2R, 6R) -2- (H-Gly-amino) -6 , 7-bis
(PamO) -4-THT-Gly-Glu-Glu-OH (Compound 9) was synthesized. Raw material (g) Reaction conditions Product (g) a) Boc-Gly-OH HGCP-4 800 mg 9a (0.13) HOBT 103 mg (0.82) WSC 146 mg DCM 8.0 ml 20 ℃ 18 hb) 9a TFA 7.1 ml 9 ( 0.74) 20 ℃ 2 h (0.56) Compound 9a: [α] _D ²⁴ -7.2 ° (c = 0.70, in chloroform) Elemental analysis value Calculated as C ₆₉ H ₁₂₅ N ₅ O ₁₆ S: C, 63.13; H, 9.60; N, 5.34; S, 2.44 Experimental value: C, 63.05; H, 9.46; N, 5.34; S, 2.38 Compound 9: (α) _D ²⁴ -14.9 ° (c = 0.56, 5% TFA-in chloroform) Elemental analysis value Calculated as C ₅₂ H ₉₃ N ₅ O ₁₄ S ・ 2H ₂ O: C, 57.81; H, 9.05; N, 6.48; S, 2.97 Experimental value: C, 57.68; H, 8.78; N, 6.31; S, 2.80

Example 10 (2R, 6R) -2- (H-Glu-Gly-amino) -6,7-bis (PamO) -4-THT-
Synthesis of Gly-Glu-Glu-OH (Compound 10) In the same manner as in Example 2, (2R, 6R) -2- (Fmoc-Gly-amino) -6,7-
Bis (PamO) -4-THT-Gly-Glu (O ^t Bu) -Glu (O ^t Bu) -O ^t Bu (Compound 10a), (2R, 6R) -2- (H-Gly-amino) -6 , 7-bis (PamO)-
4-THT-Gly-Glu (O ^t Bu) -Glu (O ^t Bu) -O ^t Bu (Compound 10b) was synthesized. Further, using compound 10b as a starting material, in the same manner as in Example 2, (2R, 6R) -2- (Fmoc-Glu (O ^t Bu) -Gly-amino) -6,7-bis (PamO) -4-THT-Gly. -Glu (O ^t Bu) -Glu (O ^t Bu) -O ^t Bu (Compound 10c), (2R, 6R) -2- (H-Glu (O ^t Bu) -Gly-amino) -6,7- Bis (PamO) -4-THT-Gly-Glu (O ^t Bu) -Glu (O ^t Bu) -O ^t Bu (Compound 10d) and (2R, 6R) -2- (H-Glu-Gly-amino) -6,7-bis (PamO) -4-THT-Gly-Glu-Glu-OH (Compound 10) was synthesized. Raw material (g) Reaction conditions Product (g) a) Fmoc-Gly-OH HGCP-4 5.00 g 10a (1.41) HOBT 643 mg (4.92) WSC 912 mg DCM 50 ml 20 ° C 14 hb) 10a Piperidine 5.0 ml 10b ( 4.70) DCM 50 ml (2.71) 20 ° C 2 h Silica gel (chloroform-methanol) 50: 1 to 20: 1 c) Fmoc-Glu (O ^t Bu)-10b 2.6 g 10c OH HOBT 312 mg (3.07) (0.56) WSC 442 mg DCM 50 ml 20 ° C 1.5 hd) 10c Piperidine 2.9 ml 10d (2.89) DCM 30 ml (2.07) 20 ° C 1.5 h Silica gel (chloroform-methanol) 50: 1 to 20: 1 e) 10d TFA 6.0 ml 10 ( 0.59) 20 ℃ 2 h (0.49) Compound 10a: [α] _D ²⁴ -8.5 ° (c = 0.91, in chloroform) Elemental analysis value Calculated as C ₇₉ H ₁₂₇ N ₅ O ₁₆ S: C, 66.13; H, 8.92; N, 4.88; S, 2.23 Experimental value: C, 65.99; H, 8.93; N, 5.00; S, 2.60 Compound 10b: (α) _D ²⁴ -6.1 ° (c = 0.81, in chloroform) Elemental analysis value C Calculated as ₆₄ H ₁₁₇ N ₅ O ₁₄ S ・ 0.5H ₂ O: C, 62.92; H, 9.74; N, 5.73; S, 2.62 Experimental value: C, 63.01; H, 9.78; N, 6.00; S, 2.54 Compound 10c: ] _D ²⁴ -7.9 ° (c = 0.67, in chloroform) Elemental analysis _{C 88 H 142 N 6 O 19} S · 0.5H 2 O Calculated: C, 64.88; H, 8.85 ;
N, 5.16; S, 1.97 experimental value: C, 64.87; H, 8.76;
N, 5.27; S, 1.99 Compound 10d: [α] _D ²⁴ -12.8 ° (c = 0.89, in chloroform) Elemental analysis value Calculated value as C ₇₃ H ₁₃₂ N ₆ O ₁₇ S ・ 0.5H ₂ O : C, 62.32; H, 9.53; N, 5.97; S, 2.28 Experimental value: C, 62.27; H, 9.44; N, 6.11; S, 2.23 Compound 10: [α] _D ²⁴ -2.4 ° (c = 0.76, 5% TFA- in chloroform) Elemental analysis value Calculated as C ₅₇ H ₁₀₀ N ₆ O ₁₇ S ・ 4H ₂ O: C, 54.97; H, 8.74; N, 6.75; S, 2.57 Experimental value: C, 54.84; H , 8.59; N, 6.60; S, 2.55

Example 11 (2R, 6R) -2- (H-Glu-Glu-Gly-amino) -6,7-bis (PamO) -4-
Synthesis of THT-Gly-Glu-Glu-OH (Compound 11) Using compound 10d as a starting material and in the same manner as in Example 2, (2R, 6R) -2- (F
moc-Glu (Ot ^B u) -Glu (O ^t Bu) -Gly-amino) -6,7-bis (PamO)
-4-THT-Gly-Glu (O ^t Bu) -Glu (O ^t Bu) -O ^t Bu (Compound 11a),
(2R, 6R) -2- (H-Glu (O ^t Bu) -Glu (O ^t Bu) -Gly-amino) -6,7-
Bis (PamO) -4-THT-Gly-Glu (O ^t Bu) -Glu (O ^t Bu) -O ^t Bu (Compound 11b) and (2R, 6R) -2- (H-Glu-Glu-Gly- amino)-
6,7-Bis (PamO) -4-THT-Gly-Glu-Glu-OH (Compound 11)
Was synthesized. Raw material (g) Reaction conditions Product (g) a) Fmoc-Glu (O ^t Bu)-10d 1.0 g 11a OH HOBT 106 mg (1.20) (0.35) WSC 151 mg DCM 25 ml 20 ° C 15 hb) 11a Piperidine 1.0 ml 11b (1.00) DCM 20 ml (0.67) 20 ° C 2 h Silica gel (chloroform-methanol) 50: 1 c) 11b TFA 5.7 ml 11 (0.57) 20 ° C 2 h (0.47) Compound 11a: [α] _D ^24- 11.1 ° (c = 0.60, in chloroform) Elemental analysis value C ₉₇ H ₁₅₇ N ₇ O ₂₂ S ・ Calculated as 0.5 H ₂ O: C, 64.21; H, 8.78; N, 5.40; S, 1.77 Experimental value: C , 64.00; H, 8.66; N, 5.24; S, 1.63 Compound 11b: (α) _D ²⁴ -10.1 ° (c = 0.55, in chloroform) Elemental analysis value C ₈₂ H ₁₄₇ N ₇ O ₂₀ S Calculated value: C , 62.21; H, 9.36; N, 6.19; S, 2.03 Experimental value: C, 62.03; H, 9.57; N, 6.10; S, 2.07 Compound 11: [α] _D ²⁴ -3.8 ° (c = 0.56, 5% Elemental analysis value Calculated as C ₆₂ H ₁₀₇ N ₇ O ₂₀ S ・ 2H ₂ O: C, 55.63; H, 8.36; N, 7.32; S, 2.40 Experimental value: C, 55.58; H, 8.12 ; N, 7.09; S, 2.39

Example 12 (2R, 6R) -2- (H-Glu-amino) -6,7-bis (PamO) -4-THT-Gly-
Synthesis of D-Glu-OH (Compound 12) As in Example 2, (2R, 6R) -2- (Fmoc-Glu (O ^t Bu) -amino)
-6,7-Bis (PamO) -4-THT-Gly--D-Glu (O ^t Bu) -O ^t Bu (Compound 12a), (2R, 6R) -2- (H-Glu (O ^t Bu ) -Amino) -6,7-bis (Pa
mO) -4-THT-Gly-D-Glu (O ^t Bu) -O ^t Bu (Compound 12b) and
(2R, 6R) -2- (H-Glu-amino) -6,7-bis (PamO) -4-THT-Gly
-D-Glu-OH (Compound 12) was synthesized. Raw material (g) Reaction conditions Product (g) a) Fmoc-Glu (O ^t Bu)-HGCP-3 1.2 g 12a OH HOBT 184 mg (1.22) (0.60) WSC 261 mg DCM 25 ml 20 ℃ 20 hb) 12a Piperidine 1.1 ml 12b (1.07) DCM 10 ml (0.70) 20 ° C 1.5 h Silica gel (chloroform-methanol) 50: 1 c) 12b TFA 5.4 ml 12 (0.54) 20 ° C 1.5 h (0.46) Compound 12a: [α] _D ²⁴ -16.3 ° (c = 0.62, in chloroform) Elemental analysis value Calculated as C ₇₇ H ₁₂₄ N ₄ O ₁₅ S: C, 67.12; H, 9.07; N, 4.07; S, 2.33 Experimental value: C, 66.97; H, 8.94; N, 4.07; S, 2.33 Compound 12b: (α) _D ²⁴ -21.2 ° (c = 0.55, in chloroform) Elemental analysis value Calculated as C ₆₂ H ₁₁₄ N ₄ O ₁₃ S: C, 64.44; H, 9.94; N, 4.85; S, 2.77 Experimental value: C, 64.13; H, 9.96; N, 4.81; S, 2.73 Compound 12: (α) _D ²⁴ + 4.5 ° (c = 0.41, 5% TFA-chloroform Medium) Elemental analysis value Calculated as C ₅₀ H ₉₀ N ₄ O ₁₃ S-4H ₂ O: C, 56.69; H, 9.32; N, 5.29; S, 3.03 Experimental value: C, 56.47; H, 8.87; N, 5.11; S, 3.04

Example 13 (2R, 6R) -2- (H-β-Ala-amino) -6,7-bis (PamO) -4-THT-G
Synthesis of ly-Gly-Gly-Glu-OH (Compound 13) HGCP-1 (43 mg) and Boc-β-Ala-OH (9 mg) DMF (1 m
l) Add TEA (0.014 ml) and DEPC (10 mg) to the solution and add 20
The mixture was stirred at ℃ for 5 hours. Water was added to the reaction solution, which was extracted with ethyl acetate, washed with a 5% (W / V) citric acid aqueous solution, a saturated sodium hydrogencarbonate aqueous solution and saturated saline, dried over anhydrous sodium sulfate, and the solvent was distilled off. The residue is subjected to silica gel column chromatography (chloroform: methanol = 93: 7).
Purified by (2R, 6R) -2- (Boc-β-Ala-amino) -6,7-bis (P
amO) -4-THT-Gly-Gly-Gly-Glu (O ^t Bu) -O ^t Bu (Compound 13a)
(25 mg, 50%) was obtained as a colorless waxy compound. IR (KBr) ν: 3290, 1735, 1630 cm ^-1 1H-NMR (CDCl ₃ ) δ: 0.88 (6H, t, J = 6.6Hz), 1.03-1.35
(48H, m), 1.41 (9H, s), 1.43 (9H, s), 1.47 (9H,
s), 1.50-1.70 (4H, m), 1.70-3.60 (14H, m), 3.85-4.
67 (11H, m), 6.50-8.30 (6H, m) To the obtained compound 13a (25 mg) was added 4N ethyl acetate (4
ml) solution was added and the mixture was stirred at 20 ° C. for 4 hours, and the solvent was evaporated to give compound 13 (21 mg, 100%) as a white powder. Melting point 98-100 ° C IR (KBr) ν: 3400, 1735, 1655 cm ^{-1 1} H-NMR (CDCl ₃ ) δ: 0.88 (6H, t, J = 6.9Hz), 1.03-1.47
(48H, m), 1.47-1.72 (4H, m), 1.72-3.20 (16H, m),
3.75-4.60 (10H, m), 5.10-5.25 (1H, m)

Example 14 (2R, 6R) -2- (H-Gly-amino) -6,7-bis (PamO) -4-THT-Gly-
Synthesis of Lys-Gly-OH (Compound 14) As in Example 1, (2R, 6R) -2- (Boc-Gly-amino) -6,7-
Bis (PamO) -4-THT-Gly-Lys (Boc) -Gly-O ^t Bu (Compound 14
a) and (2R, 6R) -2- (H-Gly-amino) -6,7-bis (PamO)-
4-THT-Gly-Lys-Gly-OH (Compound 14) was synthesized. Raw material (mg) Reaction conditions Product (mg) a) Boc-Gly-OH HGCP-6 250 mg 14a (45) HOBT 35 mg (253) WSC 49 mg DCM 10 ml 20 ° C 14 hb) 14a TFA 1.5 ml 14 ( 150) 20 ℃ 2 h (140) Compound 14a: [α] _D ²⁵ -16.2 ° (c = 0.55, in chloroform) Elemental analysis value C ₆₄ H ₁₁₈ N ₆ O ₁₄ S ・ 0.5H ₂ O Calculated value: C , 62.16; H, 9.70; N, 6.79; S, 2.59 Experimental value: C, 62.13; H, 9.61; N, 7.03; S, 2.59 Compound 14: [α] _D ²⁵ -18.3 ° (c = 0.59, in chloroform ) Elemental analysis value Calculated as C ₅₀ H ₉₄ N ₆ O ₁₀ S ・ 2TFA ・ 1.5H ₂ O: C, 52.88; H, 8.14; N, 6.85; S, 2.61 Experimental value: C, 53.01; H, 8.02; N, 6.86; S, 2.65

Example 15 Preparation of Sodium Salt of Compound 1 (Compound 15) Compound 1 (450 mg) was dissolved in 5% acetonitrile-0.5% aqueous sodium hydrogen carbonate solution (225 ml) while heating to pH.
After adjusting to 9.5, it was adsorbed on a column of Diaion HP-20 (45 ml). Resin is 5% aqueous acetonitrile (260 ml)
After washing with water, it was eluted with 40% aqueous acetonitrile (250 ml). The eluate was concentrated and freeze-dried to obtain a powder, which was suspended in acetone (5 ml).
The disodium salt of 1 (Compound 15, 352 mg) was obtained as a white powder. Elemental analysis calculated as C ₅₂ H ₉₁ N ₅ O ₁₄ SNa ₂・ 2H ₂ O: C, 55.55; H, 8.56; N, 6.23; S, 2.85; Na,
4.09 experimental value: C, 55.27; H, 8.56; N, 6.13; S, 3.35; Na,
4.6 Example 16 Preparation of sodium salt of compound 2 (compound 16) Compound 2 (400 mg) was added to 0.5% aqueous sodium hydrogen carbonate solution (20 mg).
It was dissolved in 0 ml) while heating, adjusted to pH 9.5, and then adsorbed on a column of Diaion HP-20 (80 ml). After washing the resin with water (540 ml), 40% acetonitrile water (24
It was eluted with 0 ml). The eluate was concentrated, and the powder obtained by freeze-drying was suspended in acetonitrile (5 ml), and the precipitate was collected by filtration to give the trisodium salt of compound 2 (compound
16, 352 mg) was obtained as a white powder. Elemental analysis _{C 55 H 94 N 5 O 16} SNa 3 · 3H 2 O Calculated: C, 53.43; H, 8.15 ; N, 5.66; S, 2.59; Na,
5.58 Experimental value: C, 53.39; H, 8.08; N, 5.56; S, 2.57; Na,
5.5

Example 17 Preparation of Compound 4 Sodium Salt (Compound 17) Compound 4 (300 mg) was added to 0.5% aqueous sodium hydrogen carbonate solution (15
It was dissolved in 0 ml) while heating, adjusted to pH 9.5, and adsorbed on a column of Diaion HP-20 (60 ml). After washing the resin with water (480 ml), 40% acetonitrile water (18
It was eluted with 0 ml). The eluate was concentrated and freeze-dried to suspend the resulting powder in acetonitrile (40 ml), and the precipitate was collected by filtration to give the trisodium salt of compound 4 (compound 17, 252 mg) as a white powder. It was Elemental analysis _{C 57 H 97 N 6 O 17} SNa 3 · 2H 2 O Calculated: C, 53.68; H, 7.98 ; N, 6.59; S, 2.51; Na,
5.41 Experimental value: C, 53.43; H, 7.99; N, 6.52; S, 3.03; Na,
5.3 Example 18 Preparation of Compound 6 Sodium Salt (Compound 18) Compound 6 (500 mg) was added to 0.5% aqueous sodium hydrogen carbonate solution (25
It was dissolved in 0 ml) with heating, adjusted to pH 9.5, and then adsorbed on a column of Diaion HP-20 (100 ml). After washing the resin with water (800 ml), 40% acetonitrile water (40
It was eluted with 0 ml) and 60% aqueous acetonitrile (200 ml). The eluate was concentrated, and the powder obtained by freeze-drying was suspended in acetonitrile (40 ml) and the precipitate was collected by filtration to give the disodium salt of compound 6 (compound 18, 277 mg).
Was obtained as a white powder. Elemental analysis value Calculated as C ₅₆ H ₁₀₀ N ₆ O ₁₄ SNa ₂・ 2H ₂ O: C, 56.26; H, 8.77; N, 7.03; S, 2.68; Na,
3.85 Experimental value: C, 56.10; H, 8.61; N, 7.01; S, 2.30; Na,
3.5

The structural formulas of the compounds obtained in the above Examples are shown in [Table 1]. [Table 1] Compounds Example No. Number Structural formula 1 1 (2R, 6R) -2- (H-Gly-amino) -6,7-bis (PamO) -4-THT-Glu-Gly-D-Glu- OH 2 2 (2R, 6R) -2- (H-Glu-amino) -6,7-bis (PamO) -4-THT-Glu-Gly- D-Glu-OH 3 3 (2R, 6R) -2 -(H-Gly-Glu-amino) -6,7-bis (PamO) -4-THT-Glu-Gly- D-Glu-OH 4 4 (2R, 6R) -2- (H-Glu-Gly- Amino) -6,7-bis (PamO) -4-THT-Glu-Gly-D-Glu-OH 5 5 (2R, 6R) -2- (H-Tyr-amino) -6,7-bis (PamO ) -4-THT-Glu-Gly- D-Glu-OH 6 6 (2R, 6R) -2- (H-Lys-amino) -6,7-bis (PamO) -4-THT-Glu-Gly- D-Glu-OH 7 7 (2R, 6R) -2- (H-β-Ala-amino) -6,7-bis (PamO) -4-THT-Glu-Gly- D-Glu-OH 8 8 ( 2R, 6R) -2- (H-Gly-amino) -6,7-bis (PamO) -4-THT-Glu-Gly-Glu-OH 9 9 (2R, 6R) -2- (H-Gly- Amino) -6,7-bis (PamO) -4-THT-Gly-Glu-Glu-OH 10 10 (2R, 6R) -2- (H-Glu-Gly-amino) -6,7-bis (PamO ) -4-THT-Gly-Glu- Glu-OH 11 11 (2R, 6R) -2- (H-Glu-Glu-Gly-amino) -6,7-bis (PamO) -4-THT-Gly- Glu-Glu-OH 12 12 (2R, 6R) -2- (H-Glu-amino) -6,7-bis (PamO) -4-THT-Gly-D-Glu-OH 13 13 (2R, 6R) -2- (H-β-Ala-amino) -6,7-bis (Pam O) -4-THT-Gly-Gly- Gly-Glu-OH 14 14 (2R, 6R) -2- (H-Gly-amino) -6,7-bis (PamO) -4-THT-Gly-Lys -Gly-OH ・ 2TFA 15 15 (2R, 6R) -2- (H-Gly-amino) -6,7-bis (PamO) -4-THT-Glu-Gly- D-Glu-OH ・ 2Na 16 16 (2R, 6R) -2- (H-Glu-amino) -6,7-bis (PamO) -4-THT-Glu-Gly-D-Glu-OH.3Na 17 17 (2R, 6R) -2- (H-Glu-Gly-amino) -6,7-bis (PamO) -4-THT-Glu-Gly-D-Glu-OH ・ 3Na 18 18 (2R, 6R) -2- (H-Lys-amino ) -6,7-Bis (PamO) -4-THT-Glu-Gly- D-Glu-OH ・ 2Na

Next, the biological activity of the compound (I) produced by the above method will be described. Experimental Example 1 [Table 2] shows the proliferation promoting action of the compound (I) obtained in the above-mentioned Examples on mouse bone marrow cells. Assay method: BALB / c mouse bone marrow cells 2 x 10 ⁶ / ml, 2 mM L-
Test compound in RPMI1640 medium (BW, USA) containing glutamine, 20 μg / ml gentamicin (Flow Laboratories, Scotland), 10% fetal bovine serum [Bio-Whittaker (BW), USA] Was added at an appropriate concentration and incubated at 37 ° C, 5% carbon dioxide for 3 days, followed by the MTT reduction method [Tada et al., Journal of Immunological Methods.
hods) 93, 157, 1986], the proliferation of bone marrow cells was measured. [Table 2] Mouse bone marrow cell proliferation promoting action ──────────────────────── Compound number Minimum effective concentration (MEC, ng / ml) * 1 ── ────────────────────── 1 6.25 5 6.25 ───────────────────────── * 1 The growth rate of the compound-free area is set to 1, and the concentration is shown to show 1.3 times or more growth.

Experimental Example 2 Leukocytosis of Compound (I) in Mouse [Table
3]. Assay method: 6-week-old female CDF1 / Crj mice
Cyclophosphamide dissolved in physiological saline was orally administered to 5 animals per group at a dose of 150 mg / kg, and from the next day, the compound suspended in 5% glucose was administered at the following dose for 1 day. It was subcutaneously administered once for 5 days. The day after the end of administration, ED
Approximately 100 µl of peripheral blood was collected from the orbital vein using a TA-treated glass capillary, and the white blood cell count was measured using a fully automated multi-parameter blood cell counter (Sysmex K-2000, manufactured by Toa Medical Electronics Co., Ltd.). [Table 3] Leukocytosis ────────────────────────────── Compound No. Dose White blood cell count * 1 (mg / kg / day) ────────────────────────────── 1 0.50 137 4 0.13 108 ───────────── ────────────────── * 1 Instead of cyclophosphamide, 0.2 ml of physiological saline was orally administered per 20 g of body weight, and from the next day,
The white blood cell count of mice subcutaneously administered with the same dose of 5% glucose once a day for 5 days is expressed as 100%. Cyclophosphamide was orally administered at a dose of 150 mg / kg, and from the day after that, 0.2 ml of 5% of 20 g of body weight was administered.
The mean and standard deviation of white blood cell counts of mice subcutaneously administered glucose once daily for 5 days were 34.6 +/- 9.
It was 5%.

Experimental Example 3 Acetylcholinesterase of compound (I) (hereinafter referred to as Ach
The positive cell increasing action (denoted as E) is shown in [Table 4]. Assay: AchE activity is a fluorescence assay [Ishihashi et al., Proceedings of the National Academy of
Sciences (Proceedings of the National Academy o
f Sciences, USA) Vol. 86, 5953-5957.
Page, 1989]. That is, BALB / c mouse non-adhesive bone marrow cells 1% Neutridoma (Boheringer Mannhaim (Boheringer Mannhaim (Boheringer Mannhaim), Germany) containing Iscope modified Dulbecco's medium [Gibco
1 x 10 at Gibco BRL, USA)
Suspend to a concentration of ⁶ / ml, and add a solution of Compound 1 in 25 μl aliquots to a 96-well flat-bottom plate.
After adding 00 μl each, the cells were cultured in 5% carbon dioxide at 37 ° C. for 5 days. After culturing, add 6% glutaraldehyde to
Each μl was added and fixed at 4 ° C for 30 minutes. This at 5 ° C
The supernatant was removed by centrifugation at 850 xg for 5 minutes, and the wells were washed with 100 µl of phosphate buffered saline (PBS). Then in each hole
100 μl of 0.2% polyoxyethylene-10-octylphenyl ether (POPE), 1 mM ethylenediaminetetraacetate (EDTA), 0.12M sodium chloride,
A buffer solution (pH 7.5) containing 50 mM N-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES) was injected, and a 1.6 mg / ml solution of acetylcholine iodide was added thereto.
10 μl of each was added and reacted at 33 ° C. for 3 hours. After the reaction was completed, 10 μl of each was added to 10 μl of 0.4 mM 7-diethylamino-3- (4′-maleindiphenyl) -4-methylcoumarin solution and 0.2% POPE, 1 mM EDTA, 50 m
100 μl of a buffer solution (pH 5.0) containing M sodium acetate was added and mixed, and the fluorescence intensity was measured at an excitation wavelength of 365 nm and a fluorescence wavelength of 450 nm. [Table 4] AchE positive cell increasing action ────────────────────────── Compound concentration (ng / ml) AchE increase rate ^a) ─── ─────────────────────── 1 10 1.92 ────────────────────── ───── a) AchE activity of compound-free group is shown as 1.

Experimental Example 4 Toxicity test: Trisodium salt of Compound 1 125 mg / kg
There were no deaths even after intravenous administration to mice.

As is clear from the above-mentioned biological data, the compound (I) or a salt thereof has the ability to remarkably improve the hematopoietic deficiency state, has low toxicity, and is capable of radiotherapy and chemotherapy for cancer of mammals. It can be safely used as a therapeutic or prophylactic agent for leukopenia caused by the above, as a hematopoietic promoter during bone marrow transplantation, as an immunopotentiator having a leukocyte increasing action, and as a therapeutic agent for thrombocytopenia.

Formulation Example Compound 1 (4 g) and mannitol (50 g) are dissolved in distilled water for injection (1 liter) containing 30% (W / W) of polyethylene glycol 400, sterile filtered, and placed in an ampoule. Dispensed by ml. An IV solution was prepared containing 4 mg of Compound 2 per ampoule.

[0070]

The compound (I) of the present invention or a salt thereof comprises
As a therapeutic or preventive agent for leukopenia caused by radiation therapy or chemotherapy for cancer, which has an excellent leukocytosis effect, as a hematopoietic promoter during bone marrow transplantation, and as an immunopotentiating agent having a leukocyte increase effect. Furthermore, it can be safely used as a therapeutic agent for thrombocytopenia.

Continuation of front page (51) Int.Cl. ⁶ Identification code Office reference number FI Technical display area C07K 7/06

Claims (14)

[Claims] 1. A general formula: [In the formula, R ¹ and R ² are each hydrogen or an aliphatic acyl group, R ³ and R ⁴ are each hydrogen or a protective group,
X is an amino acid sequence consisting of 1 to 10 amino acid residues which may have a protecting group, and Y is a C-terminal consisting of 1 to 5 amino acid residues which may have a protecting group. Showing the amino acid sequence to be bound] or a salt thereof. 2. The compound according to claim 1, wherein the aliphatic acyl group has 2 to 30 carbon atoms. 3. The compound according to claim 1, wherein at least one of R ¹ and R ² is an aliphatic acyl group. 4. 1 to 10 which X may have a protecting group containing at least one amino acid residue having a hydrophilic group.
An amino acid sequence consisting of individual amino acid residues.
The described compound. 5. The compound according to claim 4, wherein the amino acid having a hydrophilic group is an acidic amino acid. 6. The compound according to claim 4, wherein the amino acid having a hydrophilic group is a basic amino acid. 7. At least one of R ¹ and R ² has 2 carbon atoms.
To 18 aliphatic acyl groups, X may have a protecting group, an amino acid sequence consisting of 2 to 5 amino acid residues selected from glutamic acid, D-glutamic acid, glycine and lysine, and Y is a protecting group. The compound according to claim 1, which is an amino acid sequence consisting of 1 to 4 amino acid residues selected from glutamic acid, glycine, tyrosine, lysine, and β-alanine, which may have the following: 8. (2R, 6R) -2-glycylamino-
The compound according to claim 1, which is 6,7-bis (hexadecanoyloxy) -4-thiaheptanoyl-glutamyl-glycyl-D-glutamic acid. 9. (2R, 6R) -2-Glutamyl-glycylamino-6,7-bis (hexadecanoyloxy)-
The compound according to claim 1, which is 4-thiaheptanoyl-glutamyl-glycyl-D-glutamic acid. 10. (2R, 6R) -2-Tyrosylamino-
The compound according to claim 1, which is 6,7-bis (hexadecanoyloxy) -4-thiaheptanoyl-glutamyl-glycyl-D-glutamic acid. 11. (2R, 6R) -2-lysylamino-
The compound according to claim 1, which is 6,7-bis (hexadecanoyloxy) -4-thiaheptanoyl-glutamyl-glycyl-D-glutamic acid. 12. (2R, 6R) -2-glycylamino-
The compound according to claim 1, which is 6,7-bis (hexadecanoyloxy) -4-thiaheptanoyl-glycyl-glutamyl-glutamic acid. 13. An immunopotentiator having a leukocytosis effect, which comprises the compound according to claim 1 or a salt thereof. 14. A therapeutic agent for thrombocytopenia, which comprises the compound according to claim 1 or a salt thereof.