JPH11269195A - Peptide derivative - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new peptide derivative, and a chemokine receptor antagonist with the above peptide derivative as active ingredient. SOLUTION: This peptide derivative is shown by the formula: R<1> -X-A-Y-R<2> (A is a peptide composed of 2 or 3 amino acid residues including at least one amino acid residue selected from arginine, lysine and ornithine and also selected from the group consisting of arginine, lysine, ornithine, leucine, alanine, valine, isoleucine, glycine, glutamic acid and aspartic acid; X is either absent or, (C=O), (S=O), SO2 , NH-(C=O) or NH-(C=S); Y is either absent or, O, S or NR<3> ; R<1> and R<2> are each an alkyl residue or alkenyl, residue independently, or either R<1> or R<2> is H atom). The objective antagonist exhibits CXC and CC chemokine receptor antagonism, having excellent effect in treating or preventing AIDS, allergic diseases, inflammatory diseases and the like.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel peptide derivative and a salt thereof. Furthermore, the present invention relates to a chemokine receptor antagonist comprising this compound as an active ingredient. The compound of the present invention is used as a chemokine receptor antagonist for treating or preventing AIDS, allergic diseases, inflammatory diseases and the like.

[0002]

2. Description of the Related Art Chemokines (Chemokine, chemotactic cy)
is an abbreviation for tokine) is a group of proteins containing in this molecule four cysteines having chemotactic activity against leukemia, a CXC chemokine with one amino acid inserted between the first and second cysteines ( α chemokine) Subfamily and C adjacent to the first and second cysteines
There is the C chemokine (β chemokine) subfamily. CXC chemokines are used for T-Cellline-tropic HIV (T-HIV)
CD ₄ is to allow the entry into the positive cells. And the CC chemokine family includes RANTES, MIP-1β, MCP-1, MCP
-2, MCP-3, I-309, etc. CCR5, a second receptor for macrophage-tropic HIV (M-HIV), is present in CC chemokines. CC chemokines act alone, act on lymphocytes, eosinophils, basophils, and mast cells to migrate these cells, and also have actions such as degranulation and release of various inflammatory mediators. Interleukin 8 (IL-8) is also known as a chemokine.

[0003] A recent review (Internal Medicine 80 (2) 218-224 (19)
According to 97)), Berger et al. Found that they found that the G protein-binding receptor CXCR4 allowed entry of T cell line-tropic HIV (T-HIV) into CD4-negative cells. And this physiological ligand of CXCR4 is SDF-
It has also been found that this molecule belongs to the CXC chemokine (α-chemokine). CXCR4 is provided by Matsushima Group of Kanazawa University (currently the University of Tokyo) and SDF-1
Has already been found and cloned by the Honjo group at Kyoto University and the Kishimoto group at Osaka University. In addition, it has been reported that CC chemokines (β chemokines such as RANTES, MIP-1α, and β) detected by other groups in the United States immediately before the discovery of CXCR4 suppress HIV-1 infection. Ultimately, macrophages were found to have CCR5, and normal T cells were found to have CXCR4 along with CD4. And as the chemokine receptor antagonist, Arg Arg
Using a peptide consisting of 6 to 14 amino acids of Trp Trp Cys Xaa as an interleukin 8 antagonist (WO95 / 1670
2), non-peptide interleukin-8 antagonists containing an indole derivative as an active ingredient (JP-A-9-169729) or chemokine receptor antagonists containing a piperidine derivative (JP-A-9-249566) are known. Have been.

[0004]

DISCLOSURE OF THE INVENTION The present inventors have enthusiastically developed drugs for inhibiting diseases caused by chemokines such as interleukin 8, RANTES, MIP-1α and SDF-1. As a result of the study, it was found that a novel peptide derivative has an antagonistic effect on receptors such as IL-8, RANTES, MIP-1α, and SDF. Further, as a result of further study based on this finding, the present invention has been completed. That is, an object of the present invention is to provide a novel peptide derivative and a chemokine receptor antagonist containing the peptide derivative as an active ingredient.

[0005]

The present invention provides the following general formula:
It relates to the peptide derivative represented by (1) or a salt thereof. R ¹ -XAYR ² ... (1) wherein A contains at least one amino acid residue selected from the group consisting of arginine, lysine and ornithine;
X represents a peptide consisting of two or three amino acid residues selected from the group consisting of arginine, lysine, ornithine, leucine, alanine, valine, isoleucine, glycine, glutamic acid and aspartic acid, and X is absent or-(C = O)-,-(S = O)-, -SO _2- , -NH- (C = O)-, or
-NH- (C = S)-, Y is absent or -O-, -S- or -NR ^3- , R ¹ and R ² are each independently a carbon number
A straight-chain alkyl group, a branched alkyl group, a cyclic alkyl group, a cholesteryl group, a straight-chain alkenyl group containing 1 to 5 double bonds, a branched alkenyl group having 1 to 5 or R ¹ , R ²
Represents a hydrogen atom, a linear alkyl group having 1 to 8 carbon atoms, a branched alkyl group, a cyclic alkyl group, a linear alkenyl group, or a branched alkenyl group. R ² may have an acyloxy group at the terminal. R ³ represents a hydrogen atom, a linear alkyl group having 1 to 8 carbon atoms, a branched alkyl group, a cyclic alkyl group, a linear alkenyl group or a branched alkenyl group.

The present invention, in the general formula, R ¹ is shows a linear alkyl group having 1 to 8 carbon atoms, branched alkyl, cyclic alkyl group, a straight-chain alkenyl group or branched alkenyl group and, R ² is It is preferably a straight-chain alkyl group having 12 to 30 carbon atoms, a branched alkyl group, a cyclic alkyl group, a cholesteryl group, a straight-chain alkenyl group containing 1 to 5 double bonds or a branched alkenyl group. A is composed of A ¹ and A ² , A ¹ is an amino acid residue selected from the group consisting of D or L leucine, alanine, valine, isoleucine, glutamic acid and aspartic acid, and A ² is D or L Arginine, lysine and ornithine, it is desirable to bond the peptide using an amino acid residue selected from the group consisting of, in this case X in the general formula (1) is not present,-(C = O)-, Y is It is desirable to be -O- or -NH-. Examples of these salts include trifluoroacetate, hydrochloride, acetate, sulfate, lactate, maleate, methanesulfonate, oxalate, malonate, succinate, and fumaric acid. Salts, propionates, butyrate and the like can be mentioned.

The compound of the present invention can be produced by a known peptide synthesis method. For example, 1) condensing R ² -YH with a protected amino acid that protects the N-terminus of a predetermined amino acid (step 1), and 2) removing the amino acid N-terminal protecting group
(Steps 2) and 3) If necessary, condense amino acids sequentially (Step
3), 4) Condensation with R ¹ -XZ (Step 4) (where Z represents a halogen atom, a hydrogen atom or a hydroxyl group) 5) Removal of amino acid side chain protecting group (Step 5) be able to.

The following compounds can be exemplified as the peptide derivatives of the present invention. Acetyl-L-leucyl-L-arginine-n-dodecylamide Acetyl-L-leucyl-L-arginine-n-tridecylamide acetyl-L-leucyl-L-arginine-n-tetradecylamide acetyl-L-leucyl- L-arginine-n-hexadecylamide acetyl-L-leucyl-L-arginine stearylamide acetyl-L-leucyl-L-arginine nonadecyl ester acetyl-L-leucyl-L-arginine eicosyl ester acetyl-L-leucyl- L-arginine docosyl ester acetyl-L-leucyl-L-arginine hexacosyl ester acetyl-L-leucyl-L-arginine tricontyl ester acetyl-L-leucyl-L-arginine cholesteryl ester

[0009] The present invention relates to a chemokine receptor antagonist comprising the peptide derivative or a salt thereof as an active ingredient.
The peptide derivative or a salt thereof according to the present invention is IL-8,
It is used for the treatment or prevention of diseases caused by chemokines such as RANTES, MIP-1α and SDF-1, such as AIDS, allergic diseases and inflammatory diseases. The peptide derivative of the present invention or a salt thereof was administered to an ICR mouse in a single oral administration (dose of 500 mg / kg), and no deaths were observed after 6 days, indicating no acute toxicity. Is determined. The dose can be 0.1 to 150 mg, preferably 1 to 100 mg, per 1 kg of body weight per day, which can be administered once or in several divided doses. Oral administration is desirable, but not limited to. Parenteral, for example, injection, transdermal, enteral administration and the like can be appropriately selected. In addition, the dose can be appropriately changed depending on the symptoms of the patient.

[0010] For oral administration, the dosage form is such as powders, tablets, granules, capsules, by adding one or more pharmaceutically acceptable additives to the peptide derivative of the present invention or a salt thereof. It can be made into a suppository, an injection, an oral liquid or the like. Examples of additives include magnesium stearate, talc, lactose, dextrin, starches, methylcellulose, fatty acid glycerides, water, propylene glycol, macrogol, alcohol, crystalline cellulose, hydroxypropylcellulose, and low-substituted hydroxypropylcellulose. , Carmellose, povidone, polyvinyl alcohol, calcium stearate and the like. Further, a coloring agent, a stabilizing agent, an antioxidant, an antiseptic, a pH adjusting agent, a tonicity agent, a solubilizing agent, and / or a soothing agent can be added as necessary. Granules, tablets or capsules can also be coated with a coating base such as hydroxypropylmethylcellulose, hydroxypropylmethylcellulose phthalate and the like. It is desirable that the peptide derivative of the present invention is contained in a single dose in an amount of 0.1 to 150 mg, preferably 1 to 100 mg.

Next, a method for producing the peptide derivative of the present invention will be specifically described with reference to Example 1.

Example 1 Acetyl-L-leucyl-L-arginine-n-
Dodecyl amide (Compound No. 1) of the production method (1) acetyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6-sulphonyl -L- arginine -n- dodecyl amide (Ac- Synthesis of Leu-Arg (Pmc) -NHC ₁₂ H ₂₅ ) n-dodecylamine 124.4 mg, HOBt 90.7 mg, Ac-Leu-Arg
(pmc) 200 mg was dissolved in chloroform 2 ml and cooled to 0 ° C. There are 128.7mg of WSCI hydrochloride and 1.2ml of chloroform
, And left at 2 ° C overnight. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 15: 1) to obtain 250.7 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 7.97 (1 H, d, J = 7.5 Hz), 7.8
3 (1H, d, J = 8.0Hz), 7.18 (1H, t, J = 5.0Hz), 6.78-6.21 (3H,
br), 4.24 (1H, dt, J = 7.5,8.0Hz), 4.16-4.13 (1H, m), 3.0
8-2.94 (4H, m), 2.59 (2H, t, J = 7.0Hz), 2.47 (6H, 2s), 2.0
3 (3H, s), 1.83 (3H, s), 1.78 (2H, t, J = 6.0Hz), 1.68-1.51
(1H, m), 1.49-1.15 (32H, m), 0.88-0.82 (9H, m) MS m / z (FAB); 763 (MH ⁺ )

(2) Acetyl-L-leucyl-L-arginine
The -n- synthesis of dodecyl amide Ac-Leu-Arg (Pmc) -NHC 12 H 25 100mg was dissolved in chloroform 1 ml, was added trifluoroacetic acid 1 ml, and allowed to stand overnight at room temperature. The reaction product was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (chloroform: methanol = 8: 1) to obtain 50.3 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.00 (1H, d, J = 8.0 Hz), 7.9
3 (1H, d, J = 8.5Hz), 7.74 (1H, t, J = 5.5Hz), 7.48 (1H, t, J =
5.5Hz), 7.38-6.51 (3H, br), 4.26-4.19 (3H, m), 4.18-4.
11 (1H, m), 3.12-2.94 (4H, m), 1.84 (3H, s), 1.71-1.15 (2
5H, m), 1.10 (2H, t, J = 7.0Hz), 0.90-0.83 (9H, m) MS m / z (FAB); 497 (MH ⁺ )

[0013]

Example 2 Acetyl-L-leucyl-L-arginine-n-
Tridecyl amide (Compound No. 2) of the production method (1) acetyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6-sulphonyl -L- arginine -n- tridecyl amide ( Ac-Leu-Arg (Pmc) -NHC ₁₃ H ₂₇ ) n-tridecylamine 133.8 mg, HOBt 90.7 mg, Ac-Leu-Ar
200 mg of g (Pmc) was dissolved in 2 ml of chloroform and cooled to 0 ° C. There are 128.7mg of WSCI hydrochloride and 1.2ml of chloroform
, And left at 2 ° C overnight. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (chloroform: methanol = 15: 1) to obtain 255 mg of the title compound as a colorless syrup. ^{1 H NMR (500MHz, DMSO-} d 6) δ: 7.98 (1H, d, J = 7.5Hz), 7.8
3 (1H, d, J = 8.0Hz), 7.72 (1H, brt), 7.11-6.22 (3H, br), 4.
24 (1H, m), 4.19-4.10 (1H, m), 3.09-2.92 (4H, m), 2.59 (2
(H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.03 (3H, s), 1.83 (3H, s),
1.78 (2H, t, J = 6.5Hz), 1.68-1.52 (2H, m), 1.51-1.09 (33
H, m), 0.88-0.82 (9H, m) MS m / z (FAB); 777 (MH ⁺ )

(2) Acetyl-L-leucyl-L-arginine
Synthesis of -n-tridecylamide Ac-Leu-Arg (Pmc) -NHC ₁₃ H ₂₇ 150 mg in chloroform 1.5 ml
, And 1.5 ml of trifluoroacetic acid was added, and the mixture was left overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (chloroform: methanol = 10: 1) to obtain 11.5 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.00 (1H, d, J = 7.0 Hz), 7.9
4 (1H, d, J = 7.5Hz), 7.56 (1H, t, J = 5.5Hz), 7.50 (1H, brt),
7.41-6.52 (3H, br), 4.29-4.21 (1H, m), 4.20-4.11 (1H,
m), 3.11-2.98 (4H, m), 1.84 (3H, s), 1.71-1.11 (29H, m),
0.88-0.83 (9H, m) MS m / z (FAB); 511 (MH ⁺ )

[0015]

Example 3 Acetyl-L-leucyl-L-arginine-n-
Tetradecylamide (Compound No. 3) of the production method (1) acetyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6-sulphonyl -L- arginine -n- tetradecylamide ( Ac-Leu-Arg (Pmc) -NHC ₁₄ H ₂₉ ) n-tetradecylamine 143.3 mg, HOBt 90.7 mg, Ac-Leu-
200 mg of Arg (Pmc) was dissolved in 2 ml of chloroform and cooled to 0 ° C. 128.7 mg of WSCI hydrochloride was added to chloroform 1.2
The solution dissolved in ml was added and left at 2 ° C. for 2 days. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 15: 1) to give the title compound 256.5.
mg was obtained as a colorless syrup. ^{1 H NMR (500MHz, DMSO-} d 6) δ: 7.98 (1H, d, J = 7.5Hz), 7.8
3 (1H, d, J = 8.0Hz), 7.72 (1H, brt), 6.95-6.20 (3H, br), 4.
24-4.22 (1H, m), 3.09-2.95 (4H, m), 2.86 (2H, t, J = 7.0H
z), 2.47 (6H, 2s), 2.03 (3H, s), 1.83 (3H, s), 1.78 (2H,
t, J = 6.0Hz), 1.68-1.51 (2H, m), 1.50-1.02 (36H, m), 0.8
8-0.82 (9H, m) MS m / z (FAB); 791 (MH ⁺ )

(2) Acetyl-L-leucyl-L-arginine
Synthesis of -n-tetradecylamide Ac-Leu-Arg (Pmc) -NHC ₁₄ H ₂₉ 150 mg in chloroform 1.5 ml
, And 1.5 ml of trifluoroacetic acid was added, and the mixture was left overnight at room temperature. The reaction product was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (chloroform: methanol = 8: 1) to obtain 6.7 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.00 (1H, d, J = 8.0 Hz), 7.9
4 (1H, d, J = 8.0Hz), 7.75 (1H, t, J = 5.5Hz), 7.45 (1H, brt),
7.38-6.47 (3H, br), 4.26-4.24 (1H, m), 4.23-4.18 (1H,
m), 3.09-3.01 (4H, m), 1.84 (3H, s), 1.72-1.10 (31H, m),
0.90-0.83 (9H, m) MS m / z (FAB); 526 (MH ⁺ )

[0017]

Example 4 Acetyl-L-leucyl-L-arginine-n-
Hexadecyl amide (Compound No. 4) of the production method (1) acetyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6-sulphonyl -L- arginine -n- hexadecyl amide ( Ac-Leu-Arg (Pmc) -NHC ₁₆ H ₃₃ ) n-hexadecylamine 143.3 mg, HOBt 90.7 mg, Ac-Leu-
200 mg of Arg (Pmc) was dissolved in 2 ml of chloroform and cooled to 0 ° C. 128.7 mg of WSCI hydrochloride was added to chloroform 1.2
The solution dissolved in ml was added and left at 2 ° C. for 2 days. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 15: 1) to give the title compound 272.9
mg was obtained as a colorless syrup. ^{1 H NMR (500MHz, DMSO-} d 6) δ: 7.98 (1H, d, J = 7.5Hz), 7.8
3 (1H, d, J = 8.0Hz), 7.72 (1H, brt), 6.94-6.19 (3H, br), 4.
24 (1H, dt, J = 8.0,7.5Hz), 4.18-4.10 (1H, m), 3.05-2.94
(4H, m), 2.58 (2H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.03 (3H,
s), 1.83 (3H, s), 1.77 (2H, t, J = 6.5Hz), 1.64-1.51 (2H,
m), 1.50-1.07 (39H, m), 0.88-0.82 (9H, m) MS m / z (FAB); 819 (MH ⁺ )

(2) Acetyl-L-leucyl-L-arginine
Synthesis of -n- hexadecyl amide Ac-Leu-Arg (Pmc) -NHC 16 H 33 150mg chloroform 1.5ml
, And 1.5 ml of trifluoroacetic acid was added, and the mixture was left overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the obtained crude product was purified by silica gel column chromatography (chloroform: methanol = 8: 1) to obtain 29.8 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.00 (1H, d, J = 7.5 Hz), 7.9
4 (1H, d, J = 8.0Hz), 7.54 (1H, brt), 7.45 (1H, brt), 7.41-
6.59 (3H, br), 4.29-4.11 (2H, m), 3.16-2.98 (4H, m), 1.8
4 (3H, s), 1.73-1.14 (35H, m), 0.88-0.83 (9H, m) MS m / z (FAB); 553 (MH ⁺ )

[0019]

EXAMPLE 5 Preparation of acetyl -L- leucyl -L- arginine stearyl amide (Compound No. 5) (1) acetyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman -6 - sulfonyl -L- arginine stearyl amide (Ac-Leu-Arg (Pmc ) -NHC 18 H 37) synthesis stearylamine 181mg, HOBt 90.7mg, Ac-Leu -Arg (Pm
c) 200 mg was dissolved in 2 ml of chloroform and cooled to 0 ° C. A solution prepared by dissolving 128.7 mg of WSCI hydrochloride in 1.2 ml of chloroform was added thereto, and the mixture was allowed to stand at 2 ° C. overnight. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 15: 1) to obtain 284.1 mg of the title compound as a colorless syrup. ^{1 H NMR (500MHz, DMSO-} d 6) δ: 7.98 (1H, d, J = 7.5Hz), 7.8
3 (1H, d, J = 8.0Hz), 7.72 (1H, brt), 7.12-6.20 (3H, br), 4.
24 (1H, dt, J = 8.0,7.5Hz), 4.18-4.09 (1H, m), 3.07-2.92
(4H, m), 2.59 (2H, t, J = 7.0Hz), 2.47 (6H, 2s), 2.03 (3H,
s), 1.83 (3H, s), 1.78 (2H, t, J = 6.0Hz), 1.68-1.51 (2H,
m), 1.50-1.08 (43H, m), 0.88-0.82 (9H, m) MS m / z (FAB); 848 (MH ⁺ )

(2) Synthesis of acetyl-L-leucyl-L-arginine stearylamide Ac-Leu-Arg (Pmc) -NHC ₁₈ H ₃₇ 150 mg was added to chloroform 1.5 ml
, And 1.5 ml of trifluoroacetic acid was added, and the mixture was left overnight at room temperature. The reaction solution was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 15: 1) to obtain 15.8 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.01 (1H, d, J = 8.0 Hz), 7.9
4 (1H, d, J = 8.0Hz), 7.76 (1H, brt), 7.53 (1H, brt), 7.43-
6.55 (3H, br), 4.28-4.11 (2H, m), 3.13-2.96 (4H, m), 1.8
4 (3H, s), 1.72-1.08 (37H, m), 0.88-0.83 (9H, m) MS m / z (FAB); 581 (MH ⁺ )

[0021]

Example 6 Preparation of acetyl -L- leucyl -L- arginine nonadecyl ester (Compound No. 6) (1) acetyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman - Synthesis of 6-sulfonyl-L-arginine nonadecyl ester (Ac-Leu-Arg (Pmc) -OC ₁₉ H ₃₉ ) Dissolve 300 mg of 1-nonadecanol in 6 ml of chloroform,
294 μl triethylamine, methanesulfonyl chloride
163 μl was added and the mixture was stirred at room temperature overnight. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate to give the crude product MsO-C ₁₉ H ₃₉ and concentrated in vacuo.

On the other hand, 200 mg of Ac-Leu-Arg (Pmc)
It was dissolved in 4 ml and 0.4 ml of water, adjusted to pH 7 with a 20% aqueous cesium carbonate solution, and concentrated under reduced pressure. The residue obtained by azeotropic distillation with DMF and drying under reduced pressure was dissolved in 2 ml of DMF, and the MsO-C
The crude product ₁₉ H ₃₉ added thereto, dissolved in 4 ml of DMF, and stirred for 10 days at room temperature. Water was added to the reaction solution, and extracted with benzene / ethyl acetate = 1/2. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure.The crude product obtained was purified by silica gel column chromatography (chloroform → chloroform: methanol = 5: 1) to obtain 16.8 mg of the title compound as a colorless syrup. . ¹ H NMR (500 MHz, DMSO-d ₆ )
δ: 8.26 (1H, d, J = 7.0Hz), 7.93 (1H, d, J = 8.0Hz), 6.80-6.
30 (3H, br), 4.33-4.29 (1H, m), 4.20-4.10 (1H, m), 4.05-
3.91 (2H, m), 3.03-3.02 (2H, m), 2.58 (2H, t, J = 6.5Hz),
2.47 (6H, 2s), 2.03 (3H, s), 1.81 (3H, s), 1.77 (2H, t, J =
6.0Hz), 1.68-1.40 (47H, m), 0.89-0.84 (9H, m) MS m / z (FAB); 862 (MH ⁺ )

(2) Synthesis of acetyl-L-leucyl-L-arginine nonadecyl ester (Ac-Leu-Arg-OC ₁₉ H ₃₉₎ Ac-Leu-Arg (Pmc) -OC ₁₉ H _{39 (} 157.7 mg) was added to chloroform 1.5 ml
And trifluoroacetic acid (1.5 ml) was added thereto, followed by stirring at room temperature overnight. The reaction product was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (chloroform: methanol = 10: 1) to obtain 57.7 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.31 (1 H, d, J = 7.5 Hz), 7.9
8 (1H, d, J = 7.5Hz), 7.56 (1H, brt), 7.40-6.88 (3H, br), 4.
31-4.28 (1H, m), 4.23-4.14 (1H, m), 4.05-3.96 (2H, m),
3.14-3.02 (2H, m), 1.82 (3H, s), 1.81-1.68 (1H, m), 1.64
-1.38 (8H, m), 1.32-1.18 (32H, m), 0.89-0.83 (9H, m) MS m / z (FAB); 596 (MH ⁺ )

[0024]

EXAMPLE 7 Preparation of Acetyl -L- leucyl -L- arginine eicosyl ester (Compound No. 7) (1) acetyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman - Synthesis of 6-sulfonyl-L-arginine eicosyl ester (Ac-Leu-Arg (Pmc) -OC ₂₀ H ₄₁ ) 1-eicosanol 300 mg was dissolved in chloroform 3 ml,
280 μl triethylamine, methanesulfonyl chloride
156 μl was added, and the mixture was stirred at room temperature overnight. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate to give the crude product MsO-C ₂₀ H ₄₁ and concentrated in vacuo.

On the other hand, 150 mg of Ac-Leu-Arg (Pmc)
It was dissolved in 3 ml and 0.3 ml of water, adjusted to pH 7 with a 20% cesium carbonate aqueous solution, and concentrated under reduced pressure. The residue obtained by azeotropic distillation with DMF and concentration under reduced pressure was dissolved in 1.5 ml of DMF, and the MsO-
It was added a solution obtained by dissolving the crude product of C ₂₀ H ₄₁ in 4 ml of DMF, 7 days at room temperature and stirred for 6 days at 40 ° C.. Water was added to the reaction solution, and extracted with benzene / ethyl acetate = 1/2. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 60: 1) to give the title compound 126.5.
mg was obtained as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.26 (1H, d, J = 8.0 Hz), 7.9
3 (1H, d, J = 8.0Hz), 6.76-6.29 (3H, br), 4.33-4.29 (1H,
m), 4.18-4.11 (1H, m), 4.01-3.89 (2H, m), 3.08-2.99 (2
H, m), 2.58 (2H, t, J = 7.0Hz), 2.47 (6H, 2s), 2.03 (3H, s),
1.81 (3H, s), 1.77 (2H, t, J = 6.0Hz), 1.71-1.14 (49H, m),
0.89-0.84 (9H, m) MS m / z (FAB); 876 (MH ⁺ )

(2) Synthesis of acetyl-L-leucyl-L-arginine eicosyl ester (Ac-Leu-Arg-OC ₂₀ H ₄₁ ) Ac-Leu-Arg (Pmc) -OC ₂₀ H ₄₁ 125 mg of chloroform 1.2 ml And trifluoroacetic acid (1.2 ml) was added thereto, followed by stirring at room temperature overnight. The reaction solution was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 10: 1) to obtain 41.9 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.31 (1 H, d, J = 7.5 Hz), 7.9
8 (1H, d, J = 8.0Hz), 7.56 (1H, brt), 7.42-6.65 (3H, br), 4.
31 (1H, dt, J = 8.0,8.0Hz), 4.24-4.16 (1H, m), 4.05-3.96
(2H, m), 3.15-3.03 (2H, m), 1.83 (3H, s), 1.79-1.71 (1H,
m), 1.68-1.46 (4H, m), 1.42 (2H, t, J = 6.5Hz), 1.32-1.16
(36H, m), 0.89-0.84 (9H, m) MS m / z (FAB); 610 (MH ⁺ )

[0027]

Example 8 Preparation of Acetyl -L- leucyl -L- arginine docosyl ester (Compound No. 8) (1) acetyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman - Synthesis of 6-sulfonyl-L-arginine docosyl ester (Ac-Leu-Arg (Pmc) -OC ₂₂ H ₄₃ ) 1-docosanol 300 mg was dissolved in chloroform 6 ml, triethylamine 256 μl, methanesulfonyl chloride 1
42 μl was added, and the mixture was stirred at room temperature overnight. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate to give the crude product MsO-C ₂₂ H ₄₃ and concentrated in vacuo.

On the other hand, 200 mg of Ac-Leu-Arg (Pmc)
It was dissolved in 4 ml and 0.4 ml of water, adjusted to pH 7 with a 20% aqueous cesium carbonate solution, and concentrated under reduced pressure. The residue obtained by azeotropic distillation with DMF and concentration under reduced pressure was dissolved in 2 ml of DMF, and the MsO-C
It was added a solution obtained by dissolving the crude product of ₂₂ H ₄₃ in a mixture of DMF5.6ml and chloroform 5.6 ml, 1 day at room temperature, at 40 ° C.
Stirred for 6 days. Water was added to the reaction solution, and extracted with benzene / ethyl acetate = 1/2. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform → chloroform: methanol = 5: 1) to obtain 130 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.26 (1H, d, J = 7.5 Hz), 7.9
3 (1H, d, J = 8.0Hz), 6.75-6.28 (3H, br), 4.38-4.28 (1H,
m), 4.18-4.09 (1H, m), 3.99-3.89 (2H, m), 3.08-2.99 (2
H, m), 2.58 (2H, t, J = 7.0Hz), 2.47 (6H, 2s), 2.03 (3H, s),
1.81 (3H, s), 1.77 (2H, t, J = 6.5Hz), 1.72-1.18 (53H, m),
0.89-0.84 (9H, m) MS m / z (FAB); 904 (MH ⁺ )

(2) Synthesis of acetyl-L-leucyl-L-arginine docosyl ester (Ac-Leu-Arg-OC ₂₂ H ₄₃ ) 123.6 mg of Ac-Leu-Arg (Pmc) -OC ₂₂ H _{43 was} added to chloroform 1.2 ml
And trifluoroacetic acid (1.2 ml) was added thereto, followed by stirring at room temperature overnight. The reaction solution was concentrated under reduced pressure, and a crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 10: 1) to obtain 53.0 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.31 (1 H, d, J = 7.5 Hz), 7.9
8 (1H, d, J = 8.5Hz), 7.55 (1H, brt), 7.42-6.68 (3H, br), 4.
32-4.28 (1H, m), 4.24-4.18 (1H, m), 4.02-3.92 (2H, m),
3.11-3.06 (2H, m), 1.82 (3H, s), 1.80-1.71 (1H, m), 1.68
-1.38 (8H, m), 1.32-1.14 (38H, m), 0.89-0.84 (9H, m) MS m / z (FAB); 638 (MH ⁺ )

[0030]

Example 9 Preparation of acetyl -L- leucyl -L- arginine hexa cosyl ester (Compound No. 9) (1) acetyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman - Synthesis of 6-sulfonyl-L-arginine hexacosyl ester (Ac-Leu-Arg (Pmc) -OC ₂₆ H ₅₃ ) Dissolve 247 mg of 1-hexacosanol in 5 ml of chloroform, add 180 μl of triethylamine and 100 μl of methanesulfonyl chloride, Stirred at room temperature overnight. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate to give the crude product MsO-C ₂₆ H ₅₃ and concentrated in vacuo.

On the other hand, 200 mg of Ac-Leu-Arg (Pmc)
It was dissolved in 4 ml and 0.4 ml of water, adjusted to pH 7 with a 20% aqueous cesium carbonate solution, and concentrated under reduced pressure. The residue obtained by azeotropic distillation with DMF and drying under reduced pressure was dissolved in 2 ml of DMF, and the MsO-C
Was added a solution obtained by dissolving the crude product of ₂₆ H ₅₃ in a mixture of DMF9ml and chloroform 9 ml, was stirred for 7 days at 40 ° C.. Water was added to the reaction solution, and extracted with benzene / ethyl acetate = 1/2. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform → chloroform: methanol = 5: 1) to obtain 166.1 mg of the title compound as a colorless syrup. . ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.26 (1H, d, J = 7.0 Hz), 7.9
3 (1H, d, J = 8.0Hz), 7.11-6.26 (3H, br), 4.37-4.29 (1H,
m), 4.20-4.10 (1H, m), 4.03-3.89 (2H, m), 3.09-2.99 (2
H, m), 2.58 (2H, t, J = 7.0Hz), 2.47 (6H, 2s), 2.03 (3H, s),
1.81 (3H, s), 1.77 (2H, t, J = 6.5Hz), 1.72-1.08 (61H, m),
0.89-0.84 (9H, m) MS m / z (FAB); 960 (MH ⁺ )

(2) Synthesis of acetyl-L-leucyl-L-arginine hexacosyl ester (Ac-Leu-Arg-OC ₂₆ H ₅₃ ) 162.2 mg of Ac-Leu-Arg (Pmc) -OC ₂₆ H _{53 was} added to chloroform 1.6 ml
, 1.6 ml of trifluoroacetic acid was added, and the mixture was stirred at room temperature overnight. The reaction product was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 10: 1) to obtain 95.6 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.31 (1 H, d, J = 7.5 Hz), 7.9
8 (1H, d, J = 8.0Hz), 7.57 (1H, brt), 7.48-6.64 (3H, br), 4.
35-4.30 (1H, m), 4.22-4.21 (1H, m), 4.08-3.95 (2H, m),
3.14-3.05 (2H, m), 1.82 (3H, s), 1.81-1.71 (1H, m), 1.68
-1.39 (8H, m), 1.36-1.17 (44H, m), 1.06 (2H, t, J = 7.5Hz),
0.89-0.84 (9H, m) MS m / z (FAB); 694 (MH ⁺ )

[0033]

Example 10 Preparation of Acetyl -L- leucyl -L- arginine triacontyl ester (Compound No. 10) (1) acetyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman - Synthesis of 6-sulfonyl-L-arginine triacontyl ester (Ac-Leu-Arg (Pmc) -OC ₃₀ H ₆₁ ) 400 mg of 1-triacontanol was dissolved in 8 ml of chloroform, 254 μl of triethylamine and 141 μl of methanesulfonyl chloride were added. Stirred at room temperature overnight. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate to give the crude product MsO-C ₃₀ H ₆₁ and concentrated in vacuo.

On the other hand, 200 mg of Ac-Leu-Arg (Pmc)
It was dissolved in 4 ml and 0.4 ml of water, adjusted to pH 7 with a 20% aqueous cesium carbonate solution, and concentrated under reduced pressure. The residue obtained by azeotropic distillation with DMF and drying under reduced pressure was dissolved in 2 ml of DMF, and the MsO-C
Those of the crude product ₃₀ H ₆₁ was dissolved in a mixture of DMF 12ml and chloroform 12ml, and the mixture was stirred for 8 days at 40 ° C.. Water was added to the reaction solution, and the mixture was extracted with benzene: ethyl acetate = 1: 2. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform → chloroform: methanol = 5: 1) to obtain 230.5 mg of the title compound as a colorless syrup. . ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.26 (1H, d, J = 7.0 Hz), 7.9
3 (1H, d, J = 7.0Hz), 4.37-4.29 (1H, m), 4.18-4.09 (1H, m),
3.99-3.91 (2H, m), 3.08-3.00 (2H, m), 2.61-2.56 (2H, m
m), 2.47 (6H, 2s), 2.03 (3H, s), 1.81 (3H, s), 1.62-1.18
(69H, m), 0.88-0.84 (9H, m) MS m / z (FAB); 1016 (MH ⁺ )

[0035] (2) chloroform synthetic Ac-Leu-Arg (Pmc) -OC 30 H 61 223.6mg acetyl -L- leucyl -L- arginine triacontyl ester (Ac-Leu-Arg-OC 30 H 61) 2.2 ml
, And 2.2 ml of trifluoroacetic acid was added, followed by stirring at room temperature overnight. The reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (chloroform: methanol = 10: 1) to obtain 91.1 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.31 (1 H, d, J = 7.0 Hz), 7.9
7 (1H, d, J = 8.0Hz), 7.54 (1H, brt), 7.35-6.61 (3H, br), 4.
38-4.28 (1H, m), 4.26-4.18 (1H, m), 4.02-3.95 (2H, m),
3.48-3.39 (2H, m), 3.12-3.08 (2H, m), 1.82 (3H, s), 1.79
-1.69 (1H, m), 1.68-1.38 (8H, m), 1.32-1.12 (50H, m), 1.
06 (2H, t, J = 6.5Hz), 0.89-0.84 (9H, m) MS m / z (FAB); 750 (MH ⁺ )

[0036]

Example 11 Preparation of Acetyl-Leucyl-Arginine Cholesteryl Ester (Compound No. 11) (1) Synthesis of Acetyl-Leucyl-Arginine Cholesteryl Ester 200 mg of cholesterol was dissolved in 2 ml of DMF, and Ac-Leu-Arg (P
mc) 616.3mg, DMAP252.8mg, WSCI hydrochloride 346.6mg in DMF
Add the solution dissolved in 4 ml, and at room temperature for 24 hours at 40 ° C.
Stir for 3 days. 1N hydrochloric acid was added to the reaction solution, and the mixture was extracted with benzene: ethyl acetate = 1: 2. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 247.7 mg of a crude product of the title compound as a white solid. Dissolve this in 2.5 ml of chloroform, add 2.5 ml of trifluoroacetic acid,
Left overnight at room temperature. The crude product obtained by concentrating the reaction solution under reduced pressure was purified by silica gel column chromatography (chloroform: methanol = 10: 1) to give the title compound (19.7).
mg was obtained as a white solid. ^{1 H NMR (500MHz, DMSO-} d 6) δ: 8.31 (1H, m), 7.97 (1H, m),
7.50 (1H, brt), 7.38-6.62 (3H, br), 5.36-5.30 (1H, m),
4.49-4.41 (1H, m), 4.38-4.26 (1H, m), 4.18-4.08 (1H, m),
3.15-3.02 (2H, m), 2.29-2.12 (2H, m), 2.02-1.88 (3H,
m), 1.87-0.74 (51H, m), 0.65 (3H, s) MS m / z (FAB); 698 (MH ⁺ )

[0037]

Example 12 Acetyl-L-leucyl-L-arginine-n
- dodecyl ester (Compound No. 12) of the production method (1) acetyl -L- leucyl -N ^G -2,2,5,7,8─ pentamethyl chroman-6-sulphonyl -L- arginine -n- dodecyl ester (Ac Synthesis of -Leu-Arg (Pmc) -OC ₁₂ H ₂₅ ) 50 mg of n-dodecyl alcohol was dissolved in 1.5 ml of methylene chloride, 75 μl of triethylamine and 42 μl of methanesulfonyl chloride were added, and the mixture was stirred at room temperature for 1.5 hours. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction solution, and the mixture was extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate to give the crude product MsO-C ₁₂ H ₂₅ and concentrated in vacuo. On the other hand, Ac-Leu-Arg
Dissolve 80 mg of (Pmc) in 1.6 ml of methanol and 0.16 ml of water,
The pH was adjusted to 8 with a 20% aqueous cesium carbonate solution, and the mixture was concentrated under reduced pressure. The residue obtained by azeotropic distillation with DMF and drying under reduced pressure was treated with DMF 0.8
It was dissolved ml, and the crude product MsO-C ₁₂ H ₂₅ previously obtained, and the mixture was stirred at room temperature for 6 days. Water was added to the reaction solution, and extracted with benzene / ethyl acetate = 1/2. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure.The crude product obtained was subjected to silica gel column chromatography (chloroform / methanol = 15 /
Purification in 1) gave 32.8 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.26 (1 H, d, J = 8.0 Hz), 7.93
(1H, d, J = 8.0Hz), 6.80-6.30 (3H, br), 4.33 (1H, dt, J = 9.0
Hz, 9.0Hz), 4.20-4.12 (1H, m), 4.03-3.94 (2H, m), 3.03-
3.00 (2H, m), 2.59 (2H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.03
(3H, s), 1.82 (3H, s), 1.78 (2H, t, J = 6.5Hz), 1.74-1.19
(27H, m), 0.91-0.80 (9H, m) MS m / z (FAB); 764 (MH ⁺ )

(2) Acetyl-L-rosyl-L-arginine
Synthesis of -n- dodecyl ester (Ac-Leu-Arg-OC 12 H 25) Ac- Leu-Arg- (Pmc) -OC 12 H 25 27.3 mg of chloroform 0.3
and add 0.5 ml of trifluoroacetic acid.
After stirring for an hour, the reaction solution was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 5: 1) to obtain 15.0 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.31 (1 H, d, J = 7.5 Hz), 7.98
(1H, d, J = 8.5Hz), 7.71 (1H, brt), 7.60-6.85 (3H, br),
4.31 (1H, dt, J = 8.0Hz, 8.0Hz), 4.25-4.18 (1H, m), 4.05-
3.96 (2H, m), 3.13-3.03 (2H, m), 1.82 (3H, s), 1.80-1.71
(1H, m), 1.67-1.38 (8H, m), 1.32-1.19 (18H, m), 0.91-0.
83 (9H, m) MS m / z (FAB); 498 (MH ⁺ )

[0039]

Example 13 Acetyl-L-leucyl-L-arginine-n
- Preparation of tetradecyl ester (Compound No. 13) (1) acetyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6-sulphonyl -L- arginine -n- tetradecyl ester Synthesis of (Ac-Leu-Arg (Pmc) -OC ₁₄ H ₂₉ ) 50 mg of n-tetradecyl alcohol was dissolved in 1.5 ml of methylene chloride, 75 μl of triethylamine and 42 μl of methanesulfonyl chloride were added, and the mixture was stirred at room temperature for 1.5 hours. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction solution, and the mixture was extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate to give the crude product MsO-C ₁₄ H ₂₉ and concentrated in vacuo. On the other hand, Ac-Leu
80 mg of -Arg (Pmc) was dissolved in 1.6 ml of methanol and 0.16 ml of water, adjusted to pH 8 with a 20% aqueous cesium carbonate solution, and concentrated under reduced pressure. The residue obtained by azeotropic distillation with DMF and drying under reduced pressure
After dissolving in 0.8 ml, the crude product of MsO-C ₁₄ H ₂₉ obtained above was added, and the mixture was stirred at room temperature for 7 days. Water was added to the reaction solution, and extracted with benzene / ethyl acetate = 1/2. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform / methanol = 15: 1) to obtain 47.2 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.27 (1 H, d, J = 7.5 Hz), 7.93
(1H, d), 7.10-6.30 (3H, br), 4.36-4.30 (1H, m), 4.20-4.1
3 (1H, m), 4.04-3.93 (2H, m), 3.09-3.00 (2H, m), 2.59 (2H, m
t, J = 6.5Hz), 2.48 (6H, 2 s), 2.03 (3H, s), 1.82 (3H, s), 1.81-1.20 (37H, m), 0.91
-0.81 (9H, m) MS m / z (FAB); 792 (MH ⁺ )

(2) Acetyl-L-leucyl-L-arginine
Synthesis of -n- tetradecyl ester (Ac-Leu-Arg-OC 14 H 29) Ac- Leu-Arg- (Pmc) -OC 14 H 29 39.2mg of chloroform 0.4
and add 0.8 ml of trifluoroacetic acid.
After stirring for an hour, the reaction solution was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform / methanol = 5/1) to obtain 21.3 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.32 (1 H, d, J = 7.0 Hz), 7.98
(1H, d, J = 8.0Hz), 7.59 (1H, brt), 7.45-6.70 (3H, br),
4.30 (1H, dt, J = 8.0Hz, 8.0Hz), 4.23-4.18 (1H, m), 4.06-
3.97 (2H, m), 3.13-3.06 (2H, m), 1.82 (3H, s), 1.80-1.70
(1H, m), 1.68-1.40 (8H, m), 1.31-1.18 (22H, m), 0.90-0.
82 (9H, m) MS m / z (FAB); 526 (MH ⁺ )

[0041]

Example 14 Acetyl-L-leucyl-L-arginine-n
- Preparation of hexadecyl ester (Compound No. 14) (1) acetyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6-sulphonyl -L- arginine -n- hexadecyl ester Synthesis of (Ac-Leu-Arg (Pmc) -OC ₁₆ H ₃₃ ) 50 mg of n-hexadecyl alcohol was dissolved in 1.5 ml of methylene chloride, 75 μl of triethylamine and 42 μl of methanesulfonyl chloride were added, and the mixture was stirred at room temperature for 1.5 hours. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction solution, and the mixture was extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate to give the crude product MsO-C ₁₆ H ₃₃ and concentrated in vacuo. On the other hand, Ac-Leu-Arg
Dissolve 80 mg of (Pmc) in 1.6 ml of methanol and 0.16 ml of water,
The pH was adjusted to 8 with a 20% aqueous cesium carbonate solution and concentrated under reduced pressure. The residue obtained by azeotropic distillation with DMF and drying under reduced pressure was treated with DMF 0.8
The mixture was dissolved in ml, the crude product of MsO-C ₁₆ H ₃₃ obtained above was added, and the mixture was stirred at room temperature for 7 days. Water was added to the reaction solution, and extracted with benzene: ethyl acetate = 1: 2. The organic layer was dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The crude product obtained was subjected to silica gel column chromatography (chloroform: methanol = 15:
Purification in 1) gave 34.1 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.26 (1 H, d, J = 7.5 Hz), 7.93
(1H, d, J = 8.0Hz), 7.00-6.30 (3H, br), 4.33 (1H, dt, J = 6.0H
z), 4.21-4.12 (1H, m), 4.03-3.94 (2H, m), 3.09-3.00 (2H,
m), 2.60 (2H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.03 (3H, s), 1.8
1 (3H, s), 1.78 (2H, t, J = 6.5Hz), 1.74-1.18 (41H, m), 0.9-
0.80 (9H, m) MS m / z (FAB); 820 (MH ⁺ )

(2) Acetyl-L-leucyl-L-arginine
Synthesis of -n-hexadecyl ester (Ac-Leu-Arg-OC ₁₆ H ₃₃ ) Ac-Leu-Arg- (Pmc) -OC ₁₆ H ₃₃ 32.9 mg in chloroform 0.3
and trifluoroacetic acid (0.7 ml) was added.
After stirring for an hour, the reaction solution was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 5: 1) to obtain 12.3 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.32 (1 H, d, J = 7.5 Hz), 7.98
(1H, d, J = 8.0Hz), 7.57 (1H, brt), 7.45-6.70 (3H, br),
4.31 (1H, dt, J = 8.0Hz, 8.0Hz), 4.26-4.20 (1H, m), 4.08-
3.97 (2H, m), 3.14-3.08 (2H, m), 1.82 (3H, s), 1.80-1.70
(1H, m), 1.67-1.38 (8H, m), 1.32-1.17 (26H, m), 0.90-0.
82 (9H, m) MS m / z (FAB); 554 (MH ⁺ )

[0043]

Example 15 Acetyl-L-leucyl-L-arginine-n
-Method for producing octadecyl ester (Compound No. 15) (1) Methanesulfonyloxy-n-octadecane (MsO-C ₁₈
Synthesis of H ₃₇ ) 5.22 g of octadecyl alcohol was dissolved in 157 ml of methylene chloride, 5.38 ml of triethylamine and 2.24 ml of methanesulfonyl chloride were added, and the mixture was stirred at room temperature for 30 minutes.
A saturated aqueous sodium hydrogen carbonate solution was added to the reaction solution, and the mixture was extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the obtained crude product was purified by silica gel column chromatography (hexane: ethyl acetate = 5: 1) to obtain 6.33 g of the title compound as a white solid. ¹ H NMR (60 MHz, CDCl ₃ ) δ: 4.50-4.40 (2H, m), 3.02 (3H, s),
1.60-0.80 (35H, m),

[0044] (2) acetyl -L- leucyl -N ^G -2,2,5,7,8
-Pentamethylchroman-6-sulfonyl-L-arginine
-n-octadecyl ester (Ac-Leu-Arg (Pmc) -OC ₁₈ H ₃₇ )
Ac-Leu-Arg- (Pmc) 500 mg in methanol 10 ml and water
It was dissolved in 1 ml, adjusted to pH 8 with a 20% cesium carbonate aqueous solution, and concentrated under reduced pressure. Was azeotroped with DMF, and dried under reduced pressure to give the residue was dissolved in 5 ml of DMF, added MsO-C ₁₈ H ₃₇ 0.88g, was stirred at room temperature for 7 days. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 15: 1) to obtain 453 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.26 (1 H, d, J = 7.0 Hz), 7.93
(1H, d, J = 8.5Hz), 7.00-6.40 (3H, br), 4.33 (1H, dt, J = 9.
0Hz, 9.0Hz), 4.20-4.10 (1H, m), 4.05-3.93 (2H, m), 3.06-
3.00 (2H, m), 2.58 (2H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.03 (3
H, s), 1.82 (3H, s), 1.77 (2H, t, J = 6.5Hz), 1.74-1.19 (45
H, m), 0.91-0.80 (9H, m) MS m / z (FAB); 848 (MH ⁺ )

(3) Acetyl-L-leucyl-L-arginine
Synthesis of -n-octadecyl ester (Ac-Leu-Arg-OC ₁₈ H ₃₇ ) Ac-Leu-Arg- (Pmc) -OC ₁₈ H ₃₇ 453 mg in chloroform 4.5 ml
And add 4.5 ml of trifluoroacetic acid, and add
After stirring for an hour, the reaction solution was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 5: 1) to obtain 330 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.32 (1 H, d, J = 7.0 Hz), 7.98
(1H, d, J = 7.5Hz), 7.45 (1H, brt), 7.35 -6.60 (3H, br),
4.32 (1H, dt, J = 8.0Hz, 7.5Hz), 4.26-4.18 (1H, m), 4.05-
3.96 (2H, m), 3.11-3.05 (2H, m), 1.82 (3H, s), 1.80-1.40
(9H, m), 1.32-1.18 (30H, m), 0.92-0.80 (9H, m) MS m / z (FAB); 582 (MH ⁺ )

[0046]

Example 16 Preparation of n-dodecanoyl-L-leucyl-L-arginine methyl ester (Compound No. 16) (1) 9-Fluorenylmethoxycarbonyl- ^NG- 2,2,5,7,8
-Pentamethylchroman-6-sulfonyl-L-arginine
Synthesis of -n-octadecyl ester (Fmoc-Arg (Pmc) -OMe) The compound was dissolved in 500 mg of Fmoc-Arg (Pmc) in 10 ml of THF, 3.8 ml of trimethylsilyldiazomethane was added, and the mixture was stirred at room temperature for 20 minutes. The reaction product was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (hexane / ethyl acetate = 1/3) to obtain 520 mg of the title compound as a white solid. ^{1 H NMR (500MHz, DMSO-} d 6) δ: 7.89 (2H, d, J = 7.5Hz), 7.77 (1
H, d, J = 7.5Hz), 7.70 (2H, dd, J = 3.0Hz, 7.5Hz), 7.41 (2H, t, J
= 7.5Hz), 7.33 (2H, t, J = 7.5Hz), 6.90-6.30 (3H, br), 4.33-
4.20 (3H, m), 4.05-3.96 (1H, m), 3.61 (3H, s), 3.06-2,98 (2
H, m), 2.57 (2H, t, J = 6.5Hz), 2.48 (3H, s), 2.47 (3H, s) 2.02
(3H, s), 1.76 (2H, t, J = 6.5Hz), 1.76-1.32 (4H, m), 1.25 (6H,
2s) MSm / z (FAB): 677 (MH ⁺ )

(2) 9-Fluorenylmethoxycarbonyl
-L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman -6
-Sulfonyl-L-arginine methyl ester (Fmoc-Leu-
Synthesis of Arg (Pmc) -OMe) 469 mg of Fmoc-Arg (Pmc) -OMe was dissolved in 9.4 ml of DMF, 0.94 ml of diethylamine was added, and the mixture was stirred at room temperature for 25 minutes. The crude product obtained by concentrating the reaction solution under reduced pressure was dissolved in 6.3 ml of DMF, 266 mg of WSCI hydrochloride, 187 mg of HOBt, and 490 mg of Fmoc-Leu were added, and the mixture was added at room temperature.
Stirred for 22 hours. 1N Hydrochloric acid was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to obtain 544 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.23 (1 H, d, J = 7.5 Hz), 7.88 (2
H, d, J = 7.5Hz), 7.72 (2H, dd, J = 4.5Hz, 7.5Hz), 7.45 (1H, d, J
= 8.0Hz), 7.41 (2H, t, J = 7.5Hz), 7.31 (2H, t, J = 7.5Hz), 7.10
-6.25 (4H, br), 4.35-4.17 (3H, m), 4.12-4.07 (1H, m), 3.59
(3H, s), 3.08-2.98 (2H, m), 2.56 (2H, t, J = 7.0Hz), 2.47 (6H,
2s), 2.02 (3H, s), 1.75 (2H, t, J = 6.5Hz), 1.75-1.36 (8H, m),
1.24 (6H, 2s), 0.92-0.82 (6H, m) MS m / z (FAB); 790 (MH ⁺ )

[0048] (3) n-dodecanoyl -L- leucyl -N ^G -
2,2,5,7,8-pentamethylchroman-6-sulfonyl -L- arginine methyl ester _{(C 11 H 23 - (C} = O) -Leu-Arg (Pmc) -O
50 mg of Fmoc-Leu-Arg (Pmc) -OMe was dissolved in 3 ml of DMF, 0.3 ml of diethylamine was added, and the mixture was stirred at room temperature for 20 minutes. The crude product obtained by concentrating the reaction solution under reduced pressure was dissolved in 1 ml of methylene chloride, triethylamine (17 μl), dodecanoyl chloride (22 μl)
Was added and stirred at room temperature for 30 minutes. Water was added to the reaction solution, which was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to obtain 32.5 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.21 (1 H, d, J = 7.5 Hz), 7.86 (1
H, d, J = 8.0Hz), 6.80-6.30 (3H, br), 4.33 (1H, dt, J = 7.5Hz,
8.0Hz), 4.20-4.13 (1H, m), 3.58 (3H, s), 3.07-3.00 (2H, m),
2.59 (2H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.15-2.00 (2H, m), 2.0
3 (3H, s), 1.77 (2H, t, J = 6.5Hz), 1.72-1.36 (9H, m), 1.30-1.
17 (22H, m), 0.90-0.80 (9H, m) MS m / z (FAB); 750 (MH ⁺ )

[0049] (4) n-dodecanoyl -L- leucyl -L- arginine methyl ester _{(C 11 H 23 - (C} = O) -Leu-Arg-OMe) Synthesis _{C 11 H 23 - (C =} O) - 28.3 mg of Leu-Arg (Pmc) -OMe in chloroform
The mixture was dissolved in 0.3 ml, and 0.3 ml of trifluoroacetic acid was added, followed by stirring at room temperature for 3 hours. The reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (chloroform: methanol = 5: 1) to obtain 12.4 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.24 (1 H, d, J = 7.5 Hz), 7.90 (1
(H, d, J = 7.5Hz), 7.55 (1H, brt), 7.50-6.70 (3H, br), 4.37-4.
30 (1H, m), 4.27-4.10 (1H, m), 3.61 (3H, s), 3.12-3.06 (2H,
m), 2.18-2.02 (2H, m), 1.80-1.39 (9H, m), 1.32-1.17 (16H,
m), 0.92-0.81 (9H, m) MS m / z (FAB); 484 (MH ⁺ )

[0050]

Example 17 n- tetradecanoyl -L- leucyl -L- arginine methyl ester (Compound No. 17) Preparation (1) n- tetradecanoyl -L- leucyl -N ^G -2,2,5,7 , 8-
Pentamethyl chroman-6-sulphonyl -L- arginine methyl ester _{(C 13 H 27 - (C} = O) -Leu-Arg (Pmc) -OMe) Fmoc-Leu-Arg was synthesized by the method of Example 16 ( Pmc) -OMe50mg
Was dissolved in DMF (3 ml), diethylamine (0.3 ml) was added, and the mixture was stirred at room temperature for 20 minutes. The crude product obtained by concentrating the reaction solution under reduced pressure was dissolved in 1 ml of methylene chloride, and 17 μl of triethylamine was added.
26 μl of tetradecanoyl chloride was added, and the mixture was stirred at room temperature for 30 minutes. Water was added to the reaction solution, which was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the obtained crude product was purified by silica gel column chromatography (hexane: acetone = 2: 1) to obtain 23.2 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.21 (1 H, d, J = 7.5 Hz), 7.86 (1
H, d, J = 8.0Hz), 6.90-6.30 (3H, br), 4.33 (1H, dt, J = 7.5Hz,
8.0Hz), 4.21-4.14 (1H, m), 3.58 (3H, s), 3.06-2.99 (2H, m),
2.59 (2H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.16-2.00 (2H, m), 2.0
3 (3H, s), 1.77 (2H, t, J = 6.5Hz), 1.73-1.40 (9H, m), 1.32-1.
19 (26H, m), 0.90-0.80 (9H, m) MS m / z (FAB); 778 (MH ⁺ )

(2) n-tetradecanoyl-L-leucyl-L-
Arginine methyl ester _{(C 13 H 27 - (C} = O) -Leu-Arg-OMe)
Synthetic _{C 13 H 27 - (C =} O) -Leu-Arg (Pmc) and OMe 19 mg chloroform 0.
The mixture was dissolved in 2 ml, trifluoroacetic acid 0.2 ml was added, and the mixture was stirred at room temperature for 3 hours. The crude product obtained by concentrating the reaction solution under reduced pressure was subjected to silica gel column chromatography (chloroform:
Purification with methanol = 5: 1) gave 14.8 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.27 (1 H, d, J = 8.0 Hz), 7.90 (1
(H, d, J = 8.0Hz), 7.35 (1H, brt), 7.50-6.70 (3H, br), 4.32 (1
(H, dt, J = 8.0Hz, 8.0Hz), 4.27-4.22 (1H, m), 3.61 (3H, s), 3.1
3-3.07 (2H, m), 2.16-2.02 (2H, m), 1.80-1.40 (9H, m), 1.32-
1.18 (20H, m), 0.91-0.82 (9H, m) MS m / z (FAB); 512 (MH ⁺ )

[0052]

Example 18 n- hexadecanoyl -L- leucyl -L- arginine methyl ester (Compound No. 18) Preparation (1) n- hexadecanoyl -L- leucyl -N ^G -2,2,5,7 , 8-
Pentamethyl chroman-6-sulphonyl -L- arginine methyl ester _{(C 15 H 31 - (C} = O) -Leu-Arg (Pmc) -OMe) Fmoc-Leu-Arg was synthesized by the method of Example 16 ( Pmc) -OMe50mg
Was dissolved in DMF (3 ml), diethylamine (0.3 ml) was added, and the mixture was stirred at room temperature for 20 minutes. The crude product obtained by concentrating the reaction solution under reduced pressure was dissolved in 1 ml of methylene chloride, and 17 μl of triethylamine was added.
Hexadecanoyl chloride (29 μl) was added, and the mixture was stirred at room temperature for 30 minutes. Water was added to the reaction solution, which was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the obtained crude product was purified by silica gel column chromatography (hexane: acetone = 2: 1) to obtain 34.2 mg of the title compound as a white foam. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.21 (1 H, d, J = 7.5 Hz), 7.86 (1
H, d, J = 8.0Hz), 6.90-6.30 (3H, br), 4.33 (1H, dt, J = 7.5Hz,
8.0Hz), 4.21-4.15 (1H, m), 3.58 (3H, s), 3.06-2.99 (2H, m),
2.59 (2H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.17-2.00 (2H, m), 2.0
3 (3H, s), 1.77 (2H, t, J = 6.5Hz), 1.75-1.38 (9H, m), 1.32-1.
15 (30H, m), 0.91-0.81 (9H, m) MS m / z (FAB): 806 (MH ⁺ )

(2) n-hexadecanoyl-L-leucyl-L-
Arginine methyl ester _{(C 15 H 31 - (C} = O) -Leu-Arg-OMe)
Synthesis C ₁₅ of _{H 31 - (C = O)} -Leu-Arg (Pmc) -OMe19mg chloroform 0.
The solution was dissolved in 3 ml, and 0.3 ml of trifluoroacetic acid was added, followed by stirring at room temperature for 3 hours. The crude product obtained by concentrating the reaction solution under reduced pressure was subjected to silica gel column chromatography (chloroform:
Purification with methanol = 5: 1) gave 11.3 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.27 (1 H, d, J = 7.5 Hz), 7.90 (1
(H, d, J = 8.5Hz), 7.53 (1H, brt), 7.50-6.70 (3H, br), 4.32-4.
30 (1H, dt, J = 8.0Hz, 8.0Hz), 4.28-4.21 (1H, m), 3.61 (3H,
s), 3.15-3.05 (2H, m), 2.16-2.02 (2H, m), 1.80-1.40 (9H,
m), 1.31-1.15 (24H, m), 0.90-0.81 (9H, m) MS m / z (FAB); 540 (MH ⁺ )

[0054]

Example 19 n-octadecanoyl-L-leucyl-L-
Preparation of arginine methyl ester (Compound No. 19) (1) n-octadecanoyl -L- leucyl -N ^G -2,2,5,7,8-
Pentamethyl chroman-6-sulphonyl -L- arginine methyl ester _{(C 17 H 33 - (C} = O) -Leu-Arg (Pmc) -OMe) Fmoc-Leu-Arg was synthesized by the method of Example 16 ( Pmc) -OMe50mg
Was dissolved in DMF (3 ml), diethylamine (0.3 ml) was added, and the mixture was stirred at room temperature for 20 minutes. The crude product obtained by concentrating the reaction solution under reduced pressure was dissolved in 1 ml of methylene chloride, and 17 μl of triethylamine was added.
40 μl of hexadecanoyl chloride was added, and the mixture was stirred at room temperature for 30 minutes. Water was added to the reaction solution, which was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (hexane: acetone = 2: 1) to obtain 32.3 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.21 (1 H, d, J = 8.0 Hz), 7.86 (1
H, d, J = 8.0Hz), 6.80-6.30 (3H, br), 4.33 (1H, dt, J = 7.5Hz,
8.0Hz), 4.21-4.15 (1H, m), 3.58 (3H, s), 3.06-3.00 (2H, m),
2.59 (2H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.15-2.00 (2H, m), 2.0
3 (3H, s), 1.77 (2H, t, J = 6.5Hz), 1.73-1.36 (9H, m), 1.33-1.
17 (34H, m), 0.91-0.82 (9H, m) MS m / z (FAB); 834 (MH ⁺ )

(2) n-octadecanoyl-L-leucyl-L-
Arginine methyl ester _{(C 17 H 35 - (C} = O) -Leu-Arg-OMe)
Synthetic _{C 17 H 35 - (C =} O) chloroform -Leu-Arg (Pmc) -OMe 28.1mg
The mixture was dissolved in 0.3 ml, and 0.3 ml of trifluoroacetic acid was added, followed by stirring at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 5: 1) to obtain 11.0 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.27 (1 H, d, J = 7.0 Hz), 7.90 (1
(H, d, J = 8.0Hz), 7.60 (1H, brt), 7.50-6.30 (3H, br), 4.32 (1
(H, dt, J = 7.0Hz, 8.0Hz), 4.27-4.20 (1H, m), 3.61 (3H, s), 3.1
3-3.08 (2H, m), 2.15-2.02 (2H, m), 1.80-1.40 (9H, m), 1.32-
1.17 (28H, m), 0.92-0.83 (9H, m) MS m / z (FAB); 568 (MH ⁺ )

[0056]

EXAMPLE 20 L- leucyl -L- preparation of arginine -n- octadecyl ester (Compound No. 20) (1) N-9- fluorenyl methoxy carbonyl -L- leucyl -N ^G -2,2,5, 7,8-Pentamethylchroman-6-sulfonyl-L-arginine-n-octadecyl ester (Fmoc-Leu-
_{Arg (Pmc) -OC 18 H 37} Arg (Pmc) synthesized by the method of Example 16 of) -OC ₁₈ H ₃₇ 83.1mg of DM
Dissolved in 1.7 ml of F, 46 mg of WSCI hydrochloride, 32 mg of HOBt, Fmoc-Leu
84.8 mg was added, and the mixture was stirred at room temperature for 1 hour. 1N Hydrochloric acid was added to the reaction solution, and the mixture was extracted with a mixed solution of benzene: ethyl acetate = 1: 2. The organic layer was dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. A crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 30: 1) to obtain 154 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.22 (1 H, d, J = 7.5 Hz), 7.88 (2
H, d, J = 7.5Hz), 7.72 (2H, dd, J = 3.5Hz, 7.5Hz), 7.46 (1H, d, J
= 8.0Hz), 7.41 (2H, t, J = 7.5Hz), 7.31 (2H, 2H, t, J = 7.5Hz),
7.05-6.30 (3H, br), 4.33-4.04 (5H, m), 4.02-3.92 (2H, m),
3.08-2.97 (2H, m), 2.56 (2H, t, J = 6.5Hz), 2.46 (6H, 2s), 2.0
1 (3H, s), 1.75 (2H, t, J = 6.5Hz), 1.75-1.15 (45H, m), 0.90-
0.80 (9H, m) MS m / z (FAB); 1028 (MH ⁺ )

(2) L-leucyl-N- ^G 2,2,5,7,8-pentamethylchroman-6-sulfonyl-L-arginine-n-octadecyl ester (Leu-Arg (Pmc) -OC ₁₈ H ₃₇ ) Synthesis of Fmoc-Leu-Arg (Pmc) -OC ₁₈ H ₃₇ 123 mg in DMF2.5 ml,
0.25 ml of diethylamine was added, and the mixture was stirred at room temperature for 2 hours.
The crude product obtained by concentrating the reaction solution under reduced pressure is subjected to silica gel column chromatography (chloroform: methanol = 10: 1).
And 74.6 mg of the title compound was obtained as a colorless syrup. ^{1 H NMR (500MHz, DMSO-} d 6) δ: 8.13 (1H, d, J = 7.5Hz), 6.80-
6.30 (3H, br), 4.23-4.17 (1H, m), 4.02-3.96 (2H, m), 3.20-
3.15 (1H, m), 3.08-2.99 (2H, m), 2.58 (2H, t, J = 6.5Hz), 2.47
(6H, 2s), 2.03 (3H, s), 1.78 (2H, t, J = 6.5Hz), 1.75-1.16 (45
H, m), 0.90-0.81 (9H, m) MS m / z (FAB); 806 (MH ⁺ )

(3) Synthesis of L-leucyl-L-arginine-n-octadecyl ester (Leu-Arg-OC ₁₈ H ₃₇ ) 5.5 mg of Leu-Arg (Pmc) -OC ₁₈ H ₃₇ was dissolved in 55 μl of chloroform. 55 μl of trifluoroacetic acid was added, and the mixture was stirred at room temperature for 4.5 hours. The reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography (chloroform: methanol: water = 7: 3: 0.5) to obtain 5.1 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.70 (1 H, brd), 7.66 (1 H, br
t), 7.50-6.70 (3H, br), 4.32-4.27 (1H, m), 4.08-4.00 (2H,
m), 3.67-3.59 (1H, m), 3.13-3.08 (2H, m), 1.82-1.18 (39H,
m), 0.93-0.82 (9H, m) MS m / z (FAB); 540 (MH ⁺ )

[0059]

Example 21 Method for producing L-leucyl-L-arginine-n-hexadecylamide (Compound No. 21) (1) N-9-sulfenylenylmethoxycarbonyl-L-leucyl
-N ^G -2,2,5,7,8-Pentamethylchroman-6-sulfonyl
-L-Arginine-n-hexadecylamide (Fmoc-Leu-Arg
Synthesis of (Pmc) -NHC ₁₆ H ₃₃ ) Arg (Pmc) -NHC ₁₆ H ₃₃ 233 mg synthesized by the method of Example 16 was used.
Dissolved in 4.7 ml of DMF, WSCI hydrochloride 135 mg, HOBt 95 mg, Fmoc
248 mg of -Leu was added, and the mixture was stirred at room temperature for 1.5 hours. To the reaction solution
1N Hydrochloric acid was added, and the mixture was extracted with a mixed solution of benzene: ethyl acetate = 1: 2. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to obtain 356 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 7.90-7.75 (4H, m), 7.70 (2
H, t, J = 7.5Hz), 7.50 (1H, d, J = 8.5Hz), 7.44-7.27 (4H, m),
6.80-6.30 (3H, br), 4.36-4.16 (3H, m), 4.10-4.00 (1H,
m), 3.08-2.97 (4H, m), 2.56 (2H, m), 2.56 (2H, t, J = 6.5H
z), 2.46 (6H, 2s), 2.02 (3H, s), 1.75 (2H, t, J = 6.5Hz),
1.67-1.17 (41H, m), 0.90-0.80 (9H, m) MS m / z (FAB); 999 (MH ⁺ )

[0060] (2) L-leucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6-sulphonyl -L- arginine -n- hexadecyl amide (Leu-Arg (Pmc) -NHC 16 Synthesis of H ₃₃ ) Fmoc-Leu-Arg (Pmc) -NHC ₁₆ H ₃₃ 225 mg was dissolved in DMF 4.5 ml, diethylamine 0.45 ml was added, and the mixture was stirred at room temperature for 1 hour. The crude product obtained by concentrating the reaction solution under reduced pressure was subjected to silica gel column chromatography (chloroform: methanol =
10: 1) to give 171 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 7.98 (1H, brd), 7.87 (1H, b
rt), 6.80-6.28 (3H, br), 4.33 (1H, t, J = 5.5Hz), 4.23-4.
18 (1H, br), 4.08 (2H, dt, J = 5.5Hz), 3.05-2.98 (4H, m),
2.59 (2H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.03 (3H, s), 1.78
(2H, t, J = 6.5Hz), 1.75-1.18 (41H, m), 0.90-0.81 (9H, m) MS m / z (FAB); 777 (MH ⁺ )

(3) Synthesis of L-leucyl-L-arginine-n-hexadecylamide (Leu-Arg-NHC ₁₆ H ₃₃ ) 8.9 mg of Leu-Arg (Pmc) -NHC ₁₆ H ₃₃ was dissolved in 89 μl of chloroform. And trifluoroacetic acid (89 μl), and the mixture was stirred at room temperature for 4.5 hours. The reaction product was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform: methanol: water = 7: 3: 0.5) to obtain 2.6 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.26-8.15 (1H, br), 7.96
(1H, t, J = 5.5Hz), 7.52 (1H, brt), 7.40-6.65 (3H, br), 4.
25 (1H, br), 3.13-3.00 (4H, m), 1.75-1.19 (35H, m), 0.90
-0.81 (9H, m) MS m / z (FAB); 511 (MH ⁺ )

[0062]

Example 22 Acetyl-L-glutamyl-L-leucyl-L
-Arginine-n-octadecyl ester (Compound No. 2
Preparation of 2) (1) N-9- fluorenyl methoxy carbonyl - gamma-tert-butyl -L- glutamyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6- Sulfonyl-L-arginine-n-octadecyl ester (Fmoc-Glu (OtBu) -Leu-Arg (Pmc) -OC
Synthesis _{Leu-Arg (Pmc) -OC 18} H 37 50mg of ₁₈ H ₃₇₎ was dissolved in 1 ml DMF, WSCI hydrochloride 24 mg, HOBt17mg, the Fmoc-Glu (OtBu) 53mg was added, at room temperature
Stir for 14 hours. 1N Hydrochloric acid was added to the reaction solution, and the mixture was extracted with a mixed solution of benzene: ethyl acetate = 1: 2. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 30: 1) to obtain 64 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.25 (1 H, d, J = 6.5 Hz), 7.9
0-7.82 (3H, m), 7.71 (2H, t, J = 7.5Hz), 7.49 (1H, d, J = 8.5H
z), 7.41 (2H, t, J = 7.5Hz), 7.32 (2H, t, J = 7.5Hz), 7.10-
6.30 (3H, br), 4.36-3.91 (8H, m), 3.08-2.99 (2H, m), 2.5
7 (2H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.20 (2H, t, J = 8.0Hz),
2.02 (3H, s), 1.90-1.17 (56H, m), 0.90-0.79 (9H, m) MS m / z (FAB); 1213 (MH ⁺ )

[0063] (2) acetyl - gamma-tert-butyl -L- glutamyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6-sulphonyl -L- arginine -n- octadecyl ester (Ac-Glu (OtBu) -Leu -Arg (Pmc) -OC 18 H 37) synthesis Fmoc-Glu of (OtBu) -Leu-Arg and _{(Pmc) -OC 18 H 37 25.5mg} DMF0.5
The mixture was dissolved in ml, diethylamine (0.05 ml) was added, and the mixture was stirred at room temperature for 20 minutes. The crude product obtained by concentrating the reaction solution under reduced pressure was dissolved in 0.6 ml of pyridine, 4 μl of acetic anhydride was added, and the mixture was stirred at room temperature for 30 minutes. Water was added to the reaction solution, which was extracted with ethyl acetate.
The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to obtain 21.4 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.16 (1H, d, J = 7.0 Hz), 7.9
7 (1H, d, J = 8.0Hz), 7.87 (1H, d, J = 8.5Hz), 6.70-6.30 (3H,
br), 4.32-4.13 (3H, m), 4.02-3.93 (2H, m), 3.08-3.00 (2
H, m), 2.58 (2H, t, J = 6.5Hz), 2.47 (6H, m), 2.20 (2H, t, J =
8.0Hz), 2.03 (3H, s), 1.83 (3H, s), 1.88-1.19 (56H, m),
0.90-0.81 (9H, m) MS m / z (FAB); 1033 (MH ⁺ )

(3) Acetyl-L-glutamyl-L-leucyl
-L-Arginine-n-octadecyl ester (Ac-Glu-Leu-
Arg-OC ₁₈ H ₃₇ ) Ac-Glu (OtBu) -Leu-Arg (Pmc) -OC ₁₈ H ₃₇ 20.4 mg was dissolved in chloroform 0.2 ml, and trifluoroacetic acid 0.4 ml was added.
Stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform: methanol: water = 7: 3: 0.5) to obtain 15.7 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 12.09 (1 H, brs), 8.24 (1 H,
d, J = 7.5Hz), 7.99 (1H, d, J = 8.0Hz), 7.87 (1H, d, J = 8.5Hz),
7.59 (1H, brt), 7.50-6.70 (3H, br), 4.31 (1H, dt, J = 7.5Hz),
4.25-4.18 (1H, m), 4.04-3.97 (2H, m), 3.12-3.08 (2H, m),
2.23 (2H, t, J = 8.0Hz), 1.85 (3H, s), 1.79-1.40 (9H, m), 1.31
-1.19 (32H, m), 0.90-0.80 (9H, m) MS m / z (FAB); 711 (MH ⁺ )

[0065]

Example 23 Acetyl-L-glutamyl-L-leucyl-L
-Arginine-n-hexadecylamide (Compound No. 2
3) the production method (1) acetyl - gamma-tert-butyl -L- glutamyl -L- leucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6-sulphonyl -L- arginine -n- Hexadecylamide (Ac-Glu (Ot
Synthesis of Bu) -Leu-Arg (Pmc) -NHC ₁₆ H ₃₃ ) Dissolve 113 mg of Leu-Arg (Pmc) -NHC ₁₆ H ₃₃ in 2.2 ml of DMF and add WSCI
Hydrochloride 54 mg, HOBt 17 mg, Fmoc-Glu (OtBu) 119 mg was added,
Stirred at room temperature for 14 hours. 1N Hydrochloric acid was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate, and then concentrated under reduced pressure. The crude product obtained was dissolved in DMF (3.6 ml), diethylamine (0.36 ml) was added, and the mixture was stirred at room temperature for 30 minutes. The crude product obtained by concentrating the reaction solution under reduced pressure was dissolved in 3.6 ml of pyridine, 30 μl of acetic anhydride was added, and the mixture was stirred at room temperature for 1.5 hours.
Water was added to the reaction solution, which was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to give 78.3 mg of the title compound.
Was obtained as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.02 (1 H, d, J = 7.5 Hz), 7.9
4 (1H, d, J = 8.0Hz), 7.78 (1H, d, J = 8.0Hz), 7.67 (1H, t), 6.
80-6.30 (3H, br), 4.28-4.09 (3H, m), 3.09-2.96 (4H, m),
2.58 (2H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.21 (2H, t, J = 8.0H
z), 2.03 (3H, s), 1.83 (3H, s), 1.77 (2H, t, J = 6.5Hz), 1.
73-1.18 (52H, m), 0.90-0.80 (9H, m) MS m / z (FAB); 1004 (MH ⁺ )

(2) Acetyl-L-glutamyl-L-leucyl
-L-Arginine-n- = hexadecylamide (Ac-Glu-Leu-Ar
g-NHC ₁₆ Synthesis of _{H 33) Ac-Glu (OtBu} ) -Leu-Arg and (Pmc) -NHC ₁₆ H ₃₃ 18.9mg dissolved in chloroform 0.2 ml, of trifluoroacetic acid 0.4ml was added,
Stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform: methanol: water = 7: 3: 0.5) to obtain 21.8 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 12.09 (1 H, brs), 8.03 (1 H,
d, J = 7.5Hz), 7.91 (2H, 2d, J = 7.5Hz), 7.74 (1H, t), 7.49
(1H, brt), 7.40-6.70 (3H, br), 4.29-4.12 (3H, m), 3.12-
3.00 (4H, m), 2.24 (2H, t, J = 8.0Hz), 1.85 (3H, s), 1.74-
1.18 (37H, m), 0.90-0.80 (9H, m) MSm / z (FAB); 682 (MH ⁺ )

[0067]

Example 24 n-Hexadecanoyl-L-leucyl-L-
Arginine-n-octadecyl ester (Compound No. 2
4) the production method (1) n-hexadecanoyl -L- leucyl -N ^G -2,2,5,7,8-
Pentamethylchroman-6-sulfonyl-L-arginine-n
- octadecyl ester _{(C 15 H 31 - (C} = O) -Leu-Arg (Pmc) -O
Synthesis of C ₁₈ H ₃₇ ) 16.6 mg of Leu-Arg (Pmc) -OC ₁₈ H ₃₇ was dissolved in 0.33 ml of methylene chloride, 6 μl of triethylamine and 13 μl of hexadecanoyl chloride were added, and the mixture was stirred at room temperature for 3 hours. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to obtain 19 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.19 (1H, d, J = 7.5 Hz), 7.8
6 (1H, d, J = 8.0Hz), 6.80-6.30 (3H, br), 4.33 (1H, d, J = 7.0
Hz), 2.47 (6H, 2s), 4.19-4.10 (1H, m), 4.01-3.94 (2H,
m), 3.06-3.00 (2H, m), 2.58 (2H, t, J = 6.5Hz), 2.17 (2H, m
t, J = 7.5Hz), 2.15-2.03 (2H, m), 2.03 (3H, s), 1.77 (2H,
t, J = 6.5Hz), 1.73-1.10 (71H, m), 0.90-0.80 (12H, m) MS m / z (FAB); 1044 (MH ⁺ )

(2) n-hexadecanoyl-L-leucyl-L-
Arginine -n- octadecyl ester _{(C 15 H 31 - (C} = O) -Le
Synthesis of u-Arg-OC ₁₈ H ₃₇ ) 15.6 mg of C ₁₅ H ₃₁ (C = O) -Leu-Arg (Pmc) -OC ₁₈ H ₃₇ was dissolved in 0.2 ml of chloroform, and 0.2 ml of trifluoroacetic acid was added. Stirred at room temperature for 4 hours. The reaction product was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 5: 1) to obtain 8.9 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.26 (1H, d, J = 7.5 Hz), 7.8
9 (1H, d, J = 8.0Hz), 7.44 (1H, brt), 7.40-6.60 (3H, br), 4.
36-4.30 (1H, m), 4.24-4.18 (1H, m), 4.00 (2H, t, J = 6.0H
z), 3.13-3.07 (2H, m), 2.20-2.01 (2H, m), 1.80-1.41 (11
H, m), 1.31-1.18 (55H, m), 0.91-0.82 (12H, m) MS m / z (FAB); 778 (MH ⁺ )

[0069]

Example 25 n- hexadecanoyl -L- leucyl -L- arginine -n- hexadecyl amide (compound No. 25) of the production method (1) n- hexadecanoyl -L- leucyl -N ^G -2, 2 , 5,7,8-
Pentamethylchroman-6-sulfonyl-L-arginine-n
- hexadecyl amide _{(C 15 H 31 - (C} = O) -Leu-Arg (Pmc) -NHC
Synthesis of ₁₆ H ₃₃ ) 11.7 mg of Leu-Arg (Pmc) -NHC ₁₆ H ₃₃ was dissolved in 0.23 ml of methylene chloride, 4 μl of triethylamine and 9 μl of hexadecanoyl chloride were added, and the mixture was stirred at room temperature for 3 hours. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (chloroform: methanol = 20: 1) to obtain 6.3 mg of the title compound as a white solid. ^{1 H NMR (500MHz, DMSO-} d 6) δ: 7.94 (1H, d, J = 7.5Hz), 7.7
5-7.70 (2H, m), 6.80-6.30 (3H, br), 4.25-4.10 (2H, m), 3.
06-2.98 (4H, m), 2.58 (2H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.
15-2.04 (2H, m), 2.03 (3H, s), 1.77 (2H, t, J = 6.5Hz), 1.6
7-1.18 (61H, m), 0.90-0.80 (12H, m) MS m / z (FAB); 1015 (MH ⁺ )

(2) n-hexadecanoyl-L-leucyl-L-
Arginine -n- hexadecyl amide _{(C 15 H 31 - (C} = O) -Leu-
Synthesis C ₁₅ H ₃₁ of _{Arg-NHC 16 H 33) -} ( the C = O) -Leu-Arg ( Pmc) -NHC 15 H 33 4.9mg was dissolved in chloroform 0.05ml, trifluoroacetic acid 0.05ml addition,
Stir at room temperature for 4 hours. The crude product obtained by concentrating the reaction solution under reduced pressure was purified by silica gel column chromatography (chloroform: methanol = 7: 1) to give the title compound (1.9 mg).
Was obtained as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 7.97 (1 H, d, J = 8.0 Hz), 7.8
5 (1H, d, J = 8.0Hz), 7.75 (1H, t, J = 6.0Hz), 7.51 (1H, brt),
7.40-6.70 (3H, br), 4.24-4.04 (2H, m), 4.00 (2H, t, J = 6.
0Hz), 3.12-2.99 (4H, m), 2.16-2.05 (2H, m), 1.72-1.18
(61H, m), 0.90-0.82 (12H, m) MS m / z (FAB); 749 (MH ⁺ )

[0071]

Example 26 Acetyl-L-alanyl-L-arginine-n
-Method for producing octadecyl ester (Compound No. 26) (1) N-9-Fluorenylmethoxycarbonyl-L-alanyl
-N ^G -2,2,5,7,8-Pentamethylchroman-6-sulfonyl
-L-Arginine-n-octadecyl ester (Fmoc-Ala-Ar
g (Pmc) -OC ₁₈ H ₃₇ ) Arg (Pmc) -OC ₁₈ H ₃₇ 14 mg was dissolved in DMF 0.3 ml, and WSCI hydrochloride was dissolved.
7.7 mg, HOBt 15.5 mg, and Fmoc-Ala monohydrate 13 mg were added, and the mixture was stirred at room temperature for 3 hours. 1N Hydrochloric acid was added to the reaction solution, and the mixture was extracted with a mixed solution of benzene: ethyl acetate = 1: 2. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 15: 1) to obtain 20 mg of the title compound as a white syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.22 (1 H, d, J = 7.5 Hz), 7.8
8 (2H, d, J = 7.5Hz), 7.72 (2H, dd, J = 3.5Hz, 7.5Hz), 7.46 (1
H, d, J = 8.0Hz), 7.41 (2H, t, J = 7.5Hz), 7.31 (2H, 2H, t, J =
7.5Hz), 7.00-6.30 (3H, br), 4.33-4.03 (5H, m), 4.01-3.
93 (2H, m), 3.08-2.97 (2H, m), 2.56 (2H, t, J = 6.5Hz), 2.4
6 (6H, 2s), 2.01 (3H, s), 1.75 (2H, t, J = 6.5Hz), 1.75-1.1
5 (39H, m), 0.85 (3H, t, J = 7.0Hz) MS m / z (FAB); 986 (MH ⁺ )

(2) Acetyl-L-alanyl- ^NG- 2,2,5,7,8
-Pentamethylchroman-6-sulfonyl-L-arginine
-n-octadecyl ester (Ac-Ala-Arg (Pmc) -OC ₁₈ H ₃₇ )
19.7 mg of Fmoc-Ala-Arg (Pmc) -OC ₁₈ H ₃₇ was dissolved in 0.4 ml of DMF, 0.04 ml of diethylamine was added, and the mixture was stirred at room temperature for 10 minutes. The crude product obtained by concentrating the reaction solution under reduced pressure was treated with pyridine 0.
It was dissolved in 45 ml, acetic anhydride (4 μl) was added, and the mixture was stirred at room temperature for 1.5 hours. Water was added to the reaction solution, which was extracted with ethyl acetate.
The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 15: 1) to obtain 6.4 mg of the title compound as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.21 (1H, d, J = 7.0 Hz), 7.9
8 (1H, d, J = 8.5Hz), 7.00-6.30 (3H, br), 4.33-4.26 (1H,
m), 4.20-4.12 (1H, m), 4.03-3.93 (2H, m), 3.08-3.00 (2
H, m), 2.58 (2H, t, J = 6.5Hz), 2.47 (6H, 2s), 2.03 (3H, s),
1,81 (3H, s), 1.77 (2H, t, J = 6.5Hz), 1.72-1.15 (39H, m),
0.85 (3H, t, J = 7.0Hz) MS m / z (FAB): 806 (MH ⁺ )

(3) Acetyl-L-alanyl-L-arginine
Synthesis of -n-octadecyl ester (Ac-Ala-Arg-OC ₁₈ H ₃₇ ) 5.4 mg of Ac-Ala-Arg (Pmc) -OC ₁₈ H ₃₇ was dissolved in chloroform 0.05 ml, and trifluoroacetic acid 0.05 ml was added. Stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and the crude product obtained was purified by silica gel column chromatography (chloroform: methanol = 8: 1) to obtain 1.8 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.26 (1H, d, J = 8.0 Hz), 8.0
3 (1H, d, J = 7.5Hz), 7.46 (1H, brt), 7.40-6.60 (3H, br), 4.
31-4.20 (2H, m), 4.08-3.98 (2H, m), 3.12-3.07 (2H, m),
1.82 (3H, s), 1.80-1.46 (6H, m), 1.31-1.16 (33H, m), 0.8
5 (3H, t, J = 7.0Hz) MS m / z (FAB); 540 (MH ⁺ )

[0074]

Example 27 Acetylleucylarginine- (16-acetoxy-n-hexadecyl) ester (Compound No. 27)
(1) Synthesis of 16-acetoxy-n-hexadecanol 100 mg of 1,16-hexadecanediol was dissolved in 1.0 ml of pyridine, 37 μl of acetic anhydride was added, and the mixture was stirred at room temperature for 18 hours. A saturated aqueous ammonium chloride solution was added to the reaction solution, and the mixture was extracted with ethyl acetate. After drying the organic layer over anhydrous sodium sulfate,
The crude product obtained by concentration under reduced pressure was purified by silica gel column chromatography (hexane / ethyl acetate = 3/1),
53.1 mg of the title compound were obtained as a white solid. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 4.05 (2H, t, J = 7.0 Hz), 3.6
3 (2H, dd, J = 5.5Hz, 6.5Hz), 2.04 (3H, s), 1.67-1.52 (4H,
m), 1.39-1.22 (24H, m) MS m / z (FAB); 301 (MH ⁺ )

(2) Synthesis of acetylleucyl-N ^G -2,2,5,7,8-pentamethylchroman-6-sulfonylarginine- (16 acetoxy-n-hexadecyl) ester Ac-Leu-Arg (Pmc ) 30 mg was dissolved in DMF 0.15 ml, WSCI hydrochloride 19 mg, DMAP 6 mg were added, and AcO- (CH ₂ ) ₁₆ -OH ₁₅ mg was further added.
A solution dissolved in 0.15 ml of DMF was added, and the mixture was stirred at room temperature for 18 hours. 1N hydrochloric acid was added to the reaction solution, and benzene / ethyl acetate = 1/2
The mixture was extracted with a mixture of the above. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product obtained was purified by silica gel column chromatography (hexane / ethyl acetate = 20/1) to obtain 38 mg of the title compound as a colorless syrup. _{^{1 H NMR (500MHz, CDCl 3}} ) δ: 4.55-4.31 (2H, m), 4.15-4.01
(4H, m), 3.22-3.16 (2H, m), 2.63 (2H, t, J = 6.5Hz), 2.58 (3H,
s), 2.57 (3H, s), 2.17 (3H, s), 2.04 (6H, 2s), 1.80 (2H, t, J =
7.0Hz), 1.93-1.21 (41H, m), 0.97-0.88 (6H, m) MS m / z (FAB); 878 (MH ⁺ )

(3) Synthesis of acetylleucylarginine- (16-acetoxy-n-hexadecyl) ester Ac-Leu-Arg (Pmc) O- (CH ₂ ) ₁₆ -OAc 35 mg was added to chloroform 0.3 m
l, add 0.3 ml of trifluoroacetic acid, and add
Stirred for hours. The crude product obtained by concentrating the reaction solution under reduced pressure was dissolved in acetonitrile and washed with hexane. The acetonitrile layer was concentrated under reduced pressure to obtain 41 mg of a crude product of the title compound as a pale yellow syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.31 (1 H, d, J = 7.5 Hz), 7.9
9-7.91 (1H, m), 7.46 (1H, brt), 7.40-6.50 (3H, br), 4.40-
4.28 (1H, m), 4.25-4.16 (1H, m), 4.07-3.93 (4H, m), 3.12-
3.05 (2H, m), 1.99 (3H, s), 1.82 (3H, s), 1.80-1.18 (41H,
m), 0.92-0.84 (6H, m) MS m / z (FAB); 612 (MH ⁺ )

[0077]

Example 28 Acetylleucylarginine- (15-acetoxy-n-pentadecyl) ester (Compound No. 28)
Production method (1) acetyl-leucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6-sulfonyl arginine - (15-tert-butyldimethylsiloxy -n- pentadecyl) Synthesis of ester 1,15 -
Dissolve 3.0 g of pentadecanediol in 15 ml of DMF, and add 1.0 g of imidazole and 2.22 g of tert-butyldimethylchlorosilane.
Was added and stirred at room temperature overnight. The reaction solution was added to water, extracted with ethyl acetate, and washed with saturated saline. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 1.38 g of crude 15-tert-butyldimethylsiloxypentadecanol. On the other hand, Ac-Leu-Arg (Pmc) 421.1 mg, DMAP 172.7 mg,
271.0 mg of WSCI hydrochloride was dissolved in 4 ml of DMF and obtained there first
A solution of 170.0 mg of crude 15-tert-butyldimethylsiloxy-n-pentadecanol in 1 ml of DMF was added, and the mixture was stirred at room temperature overnight. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure.The crude product obtained was purified by silica gel column chromatography (chloroform / methanol = 40/1 → 25/1) to give 315.6 mg of the title compound as a colorless syrup Obtained. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.30-8.25 (1H, m), 7.94-
7.89 (1H, m), 6.78-6.41 (3H, br), 4.39-4.32 (1H, m), 4.19
-4.09 (1H, m), 4.03-3.92 (2H, m), 3.06-2.98 (2H, m), 2.5
7 (2H, t, J = 7.0Hz), 2.49 (6H, 2s), 2.01 (3H, s), 1.81 (3H,
s), 1.76 (2H, t, J = 7.0Hz), 1.72-1.18 (41H, m), 0.87-0.8
1 (15H, m), 0.00 (6H, s) MS m / z (FAB); 936 (MH ⁺ )

[0078] (2) acetyl isoleucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6-sulfonyl arginine - (15-hydroxy -n- pentadecyl) Synthesis of ester-acetyl isoleucyl -N ^G -2,2,5,7,8-Pentamethylchroman-6-sulfonylarginine- (15-tert-butyldimethylsiloxy-n-pentadecyl) ester 311.6 mg in THF 4.5
After dissolving in 1 ml, 1.0 ml of 1M TBAF (THF solution) was added, and the mixture was stirred at room temperature overnight. Water was added to the reaction solution, and extracted with chloroform. The organic layer was dried over anhydrous sodium sulfate, and the crude product obtained by concentration under reduced pressure was purified by silica gel column chromatography (chloroform / methanol = 20/1).
239.2 mg of the title compound were obtained as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.31-8.26 (1H, m), 7.95-
7.90 (1H, m), 6.79-6.28 (3H, br), 4.40-4.32 (1H, m), 4.30
(1H, t, J = 5.5Hz), 4.21-4.08 (1H, m), 4.03-3.89 (2H, m),
3.21-3.15 (2H, m), 3.11-2.93 (4H, m), 2.58 (2H, t, J = 6.5H
z), 2.49 (6H, 2s), 2.03 (3H, s), 1.82-1.80 (3H, m), 1.79-
1.76 (2H, m), 1.72-1.18 (37H, m), 0.95-0.83 (6H, m) MS m / z (FAB); 822 (MH ⁺ )

[0079] (3) acetyl-leucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6-sulfonyl arginine - (15-acetoxy-pentadecyl) Synthesis of acetyl ester isoleucyl -N ^G -2 After dissolving 80.0 mg of, 2,5,7,8-pentamethylchroman-6-sulfonylarginine- (15-hydroxypentadecyl) ester in 1.0 ml of pyridine,
18.8 μl of acetic anhydride was added, and the mixture was stirred at room temperature overnight. Water was added to the reaction solution, extracted with ethyl acetate, and washed with 0.5N hydrochloric acid. The organic layer was dried over anhydrous sodium sulfate, and the crude product obtained by concentration under reduced pressure was purified by silica gel column chromatography (chloroform / methanol = 15/1).
78.3 mg of the title compound were obtained as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.31-8.26 (1H, m), 7.94-
7.90 (1H, m), 6.78-6.29 (3H, br), 4.39-4.32 (1H, m), 4.21
-4.08 (1H, m), 4.02-3.89 (4H, m), 3.06-2.96 (2H, m), 2.5
8 (2H, t, J = 7.0Hz), 2.49 (6H, 2s), 2.03 (3H, s), 1.98 (3H,
s), 1.81-1.79 (3H, m), 1.79-1.76 (2H, m), 1.73-1.18 (39
H, m), 0.95-0.83 (6H, m) MS m / z (FAB); 864 (MH ⁺ )

[0080] (4) acetyl-leucyl-arginine - (15-acetoxy-pentadecyl) Synthesis of ester-acetyl-leucyl -N ^G -2,2,5,7,8- pentamethyl chroman-6-sulfonyl arginine - (15 (Acetoxypentadecyl) ester 74.5mg dissolved in chloroform 0.8ml,
0.8 ml of trifluoroacetic acid was added, and the mixture was stirred at room temperature overnight.
The crude product obtained by concentrating the reaction solution under reduced pressure is subjected to silica gel column chromatography (chloroform / methanol = 8/1).
To give 55.3 mg of the title compound as a colorless syrup. ¹ H NMR (500 MHz, DMSO-d ₆ ) δ: 8.33-8.31 (1H, m), 7.98-
7.93 (1H, m), 7.46 (1H, brt), 7.34-6.64 (3H, s), 4.36-4.
30 (1H, m), 4.22-4.18 (1H, m), 4.05-3.96 (4H, m), 3.09 (2
(H, t, J = 6.5Hz), 1.99 (3H, s), 1.83-1.82 (3H, m), 1.79-1.
24 (33H, m), 0.90-0.84 (6H, m) MS m / z (FAB); 598 (MH ⁺ )

According to the present invention, the compounds shown in Table 1 were produced according to the above-mentioned method. Where G is Gly and P is
Pro, A is Ala, L is Leu, R is Arg, T is T
hr, V for Val, K for Lys, and E for Glu.

[0082]

[Table 1]

Next, IL-8, SD of the peptide derivative of the present invention
Test examples are shown for the measurement results of F-1 and RANTES antagonistic activities.

Test Example 1 (1) Measurement of IL-8 Antagonistic Activity Human THP-1 (monocyte-derived cell line) was subjected to a binding test using ( ¹²⁵ I) IL-8 to determine the IL-8 peptide derivative of the present invention. The affinity for the receptor was examined by the following method. THP-1 cells were subcultured in RPMI-1640 medium containing 10% FCS (FBS), and 5 × 10 ⁶ cells / 400 μl of the exponentially growing cells were suspended in RPMI-1640 medium containing 0.1% bovine serum albumin. It became cloudy. Add ( ¹²⁵ I) IL-8 (final concentration 0.06 mM, specific activity 2200ci / mmo
l) was mixed with 4 µl of D'-PBS or DMSO solution of the peptide derivative of the present invention (final concentration: 100 µM). The mixture was reacted in ice for 2 hours, centrifuged, and washed 5 times with 1 ml of D-PBS. Eventually D '
-The cells were suspended in 1 ml of PBS, and the total radioactivity of the cells was measured. THP-
Specific binding amount of ( ¹²⁵ I) IL-8 to one cell (I of THP-1 cells
( ^125I ) IL-8) bound to other than the L-1 receptor was replaced with 0.12 μM IL-82 μl of TH in place of the peptide derivative of the present invention in the above procedure.
It was determined by adding to the P-1 cell suspension. ( ¹²⁵ against THP-1 cells
I) The specific binding amount of IL-8 was determined by subtracting the non-specific binding amount from the total binding amount. That is, the inhibitory rate (%) for the specific binding of THP-1 cells to IL-8 at a dose of 100 μM of the peptide derivative of the present invention was determined by the following formula. Inhibition rate (%) = [(total binding amount-non-specific binding amount)-(binding amount at the time of adding the peptide derivative of the present invention-non-specific binding amount)] ÷ (total binding amount-non-specific binding amount) x 100

(2) Measurement of SDF-1 and RANTES antagonistic activity ( ^125I ) SDF-1 and ( ^125I ) in place of ( ^125I ) IL-8 in (1) above
Antagonistic activity against these was measured using RANTES at the same concentration. The results are shown in Table 2.

[0085]

[Table 2] 阻 害 Inhibition rate (% 100 μm) Compound No. Chemical formula ─────── ─────────── IL-8 SDF-1 RANTES ────────────────────────────── 4 Ac -LRNH- (CH ₂ ) ₁₅ -CH ₃ 40 50 50 6 Ac-ELRO- (CH ₂ ) ₁₇ -CH ₃ 70 80 80 7 Ac-LRO- (CH ₂ ) ₁₈ -CH ₃ 40 50 50 8 Ac-LRO -(CH ₂ ) ₁₉ -CH ₃ 40 50 50 9 Ac-LRO- (CH ₂ ) ₂₅ -CH ₃ 40 50 50 11 Ac-LRO- (CH ₂ ) ₂₅ -CH ₃ 40 50 50 13 Ac-LRO- ( CH ₂ ) ₁₃ -CH ₃ 40 50 50 15 Ac-LRO- (CH ₂ ) ₁₇ -CH ₃ 80 70 70 16 CH _3- (CH ₂ ) ₁₀ -C (= O) -LR-OMe 40 50 50 18 CH _3- (CH ₂ ) ₁₄ -C (= O) -LR-OMe 40 50 50 21 Ac-RO- (CH ₂ ) ₁₇ -CH ₃ 40 50 50 22 LRNH- (CH ₂ ) ₁₅ -CH ₃ 60 70 70 ───────────────────────────────

As shown in this table, it was found that the peptide derivatives of the present invention exhibited antagonistic activity against IL-8, SDF-1 or RANTES, and were able to prevent diseases caused by these chemokines. .

[0087]

Test Example 2 The acute toxicity of the peptide derivative was examined. That is, 5-week-old ICR mice (male) were divided into 5 mice per group, reared after acclimation for 1 week, and the peptide derivative of Example was dissolved or dispersed in a 0.5% aqueous solution of methylcellulose, and a single oral administration (dose of 500 mg) was performed. / kg), and the number of deaths after 6 days was examined. The results are shown in Table 3. As shown in Table 3, no administration of any peptide derivatives resulted in death and no acute toxicity was confirmed.

[0088]

[Table 3] 化合物 Compound No. Number of deaths / Number of survivors ─────────────────── 2 0/5 4 0/5 6 0/5 7 0/5 8 0/5 9 0/5 11 0/5 13 0/5 15 0/5 16 0/5 18 0/5 21 0/5 22 0 / 5 ───────────────────

[0089]

Formulation Example: Compound No. 2 (10 mg), lactose (3
6mg), corn starch (150mg), microcrystalline cellulose (29m
g) and magnesium stearate (5 mg) were mixed and compressed into tablets (230 mg / tablet).

[0090]

According to the present invention, a novel peptide derivative or a salt thereof can be provided. The novel peptide derivative of the present invention or a salt thereof has CXC and CC chemokine receptor antagonism, and exhibits an excellent effect as a therapeutic or preventive agent for AIDS, allergic diseases, inflammatory diseases and the like.

[Table 1]

[Table 1]

[Table 1]

────────────────────────────────────────────────── ─── Continued on the front page (51) Int.Cl. ⁶ Identification symbol FI C07K 4/00 A61K 37/02 AED (72) Inventor Toru Yamazaki 9-22-11 Okudo, Katsushika-ku, Tokyo (72) Inventor Dewa Toshikazu 1-1-402 Minami-Kasai, Edogawa-ku, Tokyo (72) Inventor Yoshiyuki Takemura 2-18-37, Nishi-Oizumi, Nerima-ku, Tokyo (72) Inventor Shigeru Suzuki 2-11- 28 Dorumi Tsuruma 202 (72) Inventor Eiji Inoguchi 734-2 Kitanarishima-cho, Tatebayashi-shi, Gunma (72) Inventor Takeo Edamatsu 2-7-25 Higashihashimoto, Sagamihara-shi, Kanagawa

Claims (4)

[Claims] 1. A peptide derivative represented by the following general formula (1) or a salt thereof. R ¹ -XAYR ² ... (1) wherein A contains at least one amino acid residue selected from the group consisting of arginine, lysine and ornithine;
X represents a peptide consisting of two or three amino acid residues selected from the group consisting of arginine, lysine, ornithine, leucine, alanine, valine, isoleucine, glycine, glutamic acid and aspartic acid. = O)-,-(S = O)-, -SO _2- , -NH- (C = O)-, or
-NH- (C = S) - shows the, Y is absent, -O-, -S- or -NR ³ - shows a, R ^1, R ² are each independently, a carbon number 12 to 30 Represents a straight-chain alkyl group, branched alkyl group, cyclic alkyl group, cholesteryl group, straight-chain alkenyl group containing 1 to 5 double bonds or branched alkenyl group, or ^one of R ¹ and R ² is a hydrogen atom , A linear alkyl group having 1 to 8 carbon atoms, a branched alkyl group, a cyclic alkyl group, a linear alkenyl group, and a branched alkenyl group. R ² may have an acyloxy group at the terminal. R ³ represents a hydrogen atom, a linear alkyl group having 1 to 8 carbon atoms, a branched alkyl group, a cyclic alkyl group, a linear alkenyl group or a branched alkenyl group. 2. R ¹ is a linear alkyl group having 1 to 8 carbon atoms,
Shows a branched alkyl group, a cyclic alkyl group, a linear alkenyl group or a branched alkenyl group, and R ² has 12 to 30 carbon atoms.
Linear alkyl group, branched alkyl group, cyclic alkyl group,
The peptide derivative or a salt thereof according to claim 1, which represents a cholesteryl group, a linear alkenyl group containing 1 to 5 double bonds or a branched alkenyl group. 3. A comprises A ¹ and A ² , wherein A ¹ is an amino acid residue selected from the group consisting of leucine, alanine, valine, isoleucine, glutamic acid and aspartic acid of D or L, and A ² is D Or the peptide derivative or salt thereof according to claim 1 or 2, which represents an amino acid residue selected from the group consisting of arginine, lysine and ornithine in L 1. 4. X is absent or represents-(C = O)-;
Represents a peptide derivative or a salt thereof according to any one of claims 1 to 3, which represents -O- or -NH-.