JPH068313B2 - Novel acylglutamyl lysine derivative - Google Patents

Description

The present invention relates to a novel acylglutamyl lysine derivative, and more particularly to a novel acyl glutamyl lysine derivative having excellent infection protective effect, a pharmaceutically acceptable salt thereof, and a method for producing the compound. And a pharmaceutical composition containing the compound or a pharmaceutically acceptable salt thereof.

The inventors of the present invention have synthesized many peptide compounds and applied for patents (see, for example, JP-A-56-45449). However, as a result of earnest research, a novel description not specifically described in the above-mentioned publication has been made. The present invention was completed by synthesizing various compounds and found that the compounds have an excellent infection-protecting effect.

The novel acylglutamyl lysine derivative of the present invention has the following formula
It is indicated by (I).

(In the formula, R ¹ means palmitoyl, R ² means hydrogen or benzyl, and R ₃ means hydrogen or benzyloxycarbonyl, respectively.) A pharmaceutically acceptable salt of the novel acylglutamyllysine derivative represented by the above formula (I). Examples thereof include alkali metal salts (eg sodium salt, potassium salt etc.), alkaline earth metal salts (eg calcium salt etc.), ammonium salts, organic amine salts (eg ethanolamine salt, triethylamine salt, dicyclohexylamine salt). Etc.) and the like, salts with inorganic or organic bases, methanesulfonates, hydrochlorides, sulfates, nitrates, phosphates and other acid addition salts with organic or inorganic acids.

The object compound (I) of the present invention can be synthesized by various methods detailed below.

(1) Method 1: Peptide bond formation reaction: (2) Method 2: Method for removing protecting group: The above method will be described in detail below.

[1] Peptide bond forming reaction: This reaction can be performed as follows. That is, the carboxyl group of the compound (1) or a salt thereof is first converted into an active substance such as an acid halide, an acid azide, an acid anhydride or a mixed acid anhydride, an active ester by a conventional method, and then,
The target compound (I ^a ) can be synthesized by reacting with the compound (2), and the compound (1) or a salt thereof and the compound (2) or a salt thereof are usually used as a condensing agent such as N, N- The target compound (I ^a ) can also be synthesized by directly reacting in the presence of dicyclohexylcarbodiimide and the like.

This reaction proceeds smoothly in a solvent such as methylene chloride, chloroform, tetrahydrofuran, dioxane, ethyl acetate, methanol, ethanol and water in the range of -20 ° C to room temperature. The reaction in the presence of the condensing agent is usually carried out under anhydrous conditions and mild conditions.

[2] Protective Group Elimination Reaction: This reaction is carried out by subjecting compound (I ^a ) or a salt thereof to an amino and carboxy protective group elimination reaction.
(I ^b ) or a salt thereof.

[2-1] Amino-protecting group elimination reaction: This reaction is a conventional reduction method such as catalytic reduction, liquid ammonia alkali metal method, acid method, zinc acid method, base method, hydrazine method or the like. Done by the method. The reaction temperature is usually from cooling to room temperature.

[2-2] Desorption reaction of carboxy protecting group: This reaction is carried out by a usual method such as hydrolysis or reduction. Hydrolysis normally proceeds smoothly in a solvent in the presence of an acid or a base under relatively mild conditions such as cooling or heating. The reduction includes chemical reduction and catalytic reduction, and is performed according to a conventional method. The reaction is usually performed in a solvent under relatively mild conditions such as cooling and heating.

The compounds (1) and (2) used as raw materials in the present invention are novel compounds and can be produced by the methods shown in the production examples below.

The object compound [I] and the starting compound of the present invention contain one or more isomers due to an asymmetric carbon atom in the molecule,
All such isomers are also included in the scope of the present invention.

It was found that the novel acylglutamyl lysine derivative (I) and its pharmaceutically acceptable salt provided by this invention have an excellent protective effect against experimental infectious diseases.

Therefore, the novel acylglutamyl lysine derivative (I) and its pharmaceutically acceptable salts are useful as a treatment for infectious diseases caused by pathogenic microorganisms, especially Gram-negative bacteria, Gram-positive bacteria and fungi.

Next, with respect to representative examples of the object compound (I) of the present invention, the protective effect against infection will be shown by the following test examples.

Test example: To know the protective effect of experimental infections in mice,
The test compound was dissolved in physiological saline and diluted to prepare a test solution having a specified concentration.

Male ICR-strain mice (4 weeks old) were grouped in groups of 10 mice. E. coli NO.2 on Tripuricase Soy Agar
Incubate 2 at 37 ° C overnight, suspend in physiological saline for 2.6
A suspension having a microbial cell concentration of × 10 ⁹ CFU / m was obtained. To each mouse, 8.7 × 10 ⁷ CFU of cells was intraperitoneally administered. For test compounds, 10 per group
Various amounts were intraperitoneally administered to 4 mice in advance for 4 days. The survival rate was calculated from the number of surviving animals on the third day after the administration of the bacterial cells, and the results are shown in the table.

The pharmaceutical composition of the present invention is provided as various pharmaceutical preparations, for example, a solid drug, a semi-solid drug, and a liquid drug, and these are an organic or inorganic carrier or excipient suitable for external use, internal use or topical application. It contains the active substance of the present invention.
The active ingredient is then used in combination with a harmless and pharmaceutically acceptable auxiliary ingredient for forming tablets, pellets, capsules, suppositories, solutions, emulsions, suspensions or other suitable forms. . As such an auxiliary ingredient,
Ingredients effectively used in the production of solid preparations, semi-solid preparations or liquids such as water, glucose, lactose, gelatin, mannitol, starch paste, magnesium silicate, corn starch, keratin, colloidal silica, potato starch, urea, etc. Are exemplified, and a stabilizer, a bulking agent, a coloring agent, a fragrance and the like can be supplementarily added. In addition, an antiseptic agent or a bactericidal agent may be added to the pharmaceutical composition of the present invention in order to maintain the activity of the active ingredient. The active ingredient content of the composition should be such that the active ingredient is contained in an amount sufficient to exert a desired therapeutic effect on the progress and condition of the disease.

In applying this composition to the human body, intravenous administration,
Intramuscular administration or oral administration is desired. The effective dose of the target substance of the present invention varies depending on the age and symptoms of the target patient, but is usually 0.1 to 1 per 1 kg of body weight for humans or animals.
The daily dose is 00 mg, and the content in the preparation is 50 mg of the drug.
It is generally 100 mg, 250 mg, or 500 mg.

Examples of the present invention will be described below. In the following examples, starting materials and target materials are represented by the following abbreviations.

Ala: Alanyl Glu: Glutamyl Gly: Glycyl Z: Benzyloxycarbonyl Bzl: Benzyl Su: N-hydroxysuccinimide Lys: Lysyl Reference Example 1 (1) Step 1 HD-GluOBzl (1) (7.11g) in methylene chloride (160m
) And methanol (60 m) suspension are charged with trimethylamine (6.06 g) and behenic acid succinimide ester (2) (13.11 g). The mixture is stirred at room temperature for 16 hours. After evaporating the solvent under reduced pressure, 6N hydrochloric acid (10 m) and water (400 m) are added to the residue. The obtained crude crystals are collected, washed with water and dried, and the obtained crystals are dissolved in hot ethyl acetate (200 m) and dried. After passing, the solution is diluted with diisopropyl ether and left for 2 hours at room temperature. The precipitate obtained is collected and washed with ethyl acetate to give crystals. The crystals were recrystallized from ethyl acetate to give behenoyl-D-GluoBzl (3).
(8.84 g) are obtained.

mp92-94.5 ° C IR (Nujol): 3300, 1740, 1720, 1650, 1540 cm ^-1 NMR (CDCl ₃ ) δ: 0.70-2.65 (47H, m), 4.50-4.98 (1H, m), 5.2
0 (2H, s), 6.60 (1H, d, J = 7Hz), 7.33 (5H, s), 9.93 (1H, broad
s) (2) Process 2 To a solution of behenoyl-D-GluoBzl (3) (8.41 g) in chloroform (160 m) was added N-hydroxysuccinimide (1.90 g) and dicyclohexylcarbodiimide (3.25 g) at 11 ° C. 18 at 10 ° C
Leave for hours. The precipitate obtained is removed and the liquid is concentrated under reduced pressure. Diisopropyl ether was added to the residue, and the obtained crystals were collected to give behenoyl-D-Glu (OSu) oBzl.
(4) (9.80 g) is obtained.

IR (Nujol): 3320,1810,1780,1740,1645cm ^-1 Production example (1) Step 1 Palmitoyl-D-GluoBzl (3) is obtained in substantially the same manner as in the method of Step 1 of Reference Example 1.

mp 81.5-82.5 ° C IR (Nujol): 3300, 1750, 1710, 1650 cm ^-1 NMR (CDCl ₃ ) δ: 0.91 (3H, m), 1.10-2.65 (32H, m), 4.48-4.9
2 (1H, m), 5.15 (2H, s), 6.70 (1H, d, J = 8Hz), 7.35 (5H, s), 1
0.50 (1H, s) (2) Process 2 Palmitoyl-D-Glu (OSu) OBzl (4) is obtained in substantially the same manner as in the method of Step 2 of Reference Example 1.

IR (Nujol): 3320,1810,1775,1745,1740,1650cm ^-1 NMR (CDCl ₃ ): 0.90 (3H, s), 1.10-2.77 (32H, m), 2.83 (4H,
s), 4.69-4.93 (1H, m), 5.19 (2H, s), 6.47 (1H, d, J = 8Hz), 7.
35 (5H, s) Reference example 2 (1) Process 1 Triethylamine (2.73 g) and behenoyl-D-Glu (OSu) OB were added to a suspension of L-Lys (ε-Z) -D-AlaOH (2) in methanol (100 m) and chloroform (100 m).
Add zl (1) (8.86 g). After stirring for 4 hours at room temperature, the reaction mixture is concentrated under reduced pressure. 1N Hydrochloric acid (30 m) and water (150 m) are added to the concentrate, and the obtained crystalline solid is collected and washed with water. Drying the resulting crude crystals,
When washed with hot diisopropyl ether, behenoyl-
γ-D-Glu (α-OBzl) -L-Lys (ε-Z) -D-AlaOH (3) (11.
00g) is obtained.

mp157-1260 ° C [α] _D = -7.84 ° (c = 0.82, acetic acid) IR (Nujol): 3280,1725,1680,1630cm ^-1 NMR (DMSO-d ₆ ) δ: 0.89 (3H, m), 1.00〜2.42 (53H, m), 2.76
~ 3.15 (2H, m), 4.01 ~ 4.46 (2H, m), 5.07 (2H, s), 5.17 (2H,
s), 7.42 (10H, s), 7.87 to 8.35 (4H, m) (2) Step 2 Behenoyl-γ-D-Glu (α-OBzl) -L-Lys (ε-Z) -D-Al
Dissolve aOH (3) (1.0g) in acetic acid (20m) and add 10%
Hydrogenate in the presence of Paradium Black (200 mg) in a hydrogen stream at 1 atm. After removing the catalyst, acetic acid is distilled off. The residue was collected and washed with diisopropyl ether, and the resulting crude crystals were washed with water and dried to give behenoyl-γ-DG.
This gives lu (α-OH) -L-Lys-D-AlaOH (4) (624 mg).

mp 200-204 ° C (decomposition) [α] _D = -15.0 (c = 0.31, acetic acid) IR (Nujol): 3290,1720,1630cm ^-1 Elemental analysis C ₃₆ H ₆₈ N ₄ O ₇ calculated value c: 64.63, H: 10.25, N: 8.38 Measured value c: 64.02, H: 10.08, N: 8.39 Example (1) Step 1 By a method substantially similar to the method of Step 1 of Reference Example 2,
Palmitoyl-γ-D-Glu (α-OBzl) -L-Lys (ε-
Z) -D-AlaOH (3) is obtained.

mp153-156 ° C [α] _D = -7.84 (c = 1.03, acetic acid) IR (Nujol): 3300, 1730, 1690, 1630 cm ^-1 NMR (CDCl ₃ ) δ: 0.87 (3H, m), 1.00-1.65 (41H, m), 2.75-3.4
5 (2H, m), 4.15-4.83 (3H, m), 5.05 (2H, s), 5.12 (2H, s), 7.30
(10H, s) Elemental analysis C ₄₅ H ₆₈ N ₄ O ₉ Calculated value c: 65.76, H: 8.40, N: 7.42 Measured value c: 65.89, H: 8.41, N: 7.47 (2) Step 2 Palmitoyl-γ-D-Glu (α-OH) -L-Lys-D-AlaOH (4) is obtained in substantially the same manner as in the method of Step 2 of Reference Example 2.

mp 193-199 ° C.

[Α] _D = -16.45 (c = 0.5, acetic acid) IR (Nujol): 3290,720 (shoulder), 1700,1635cm ^-1 NMR (D ₂ O-NaOD) δ: 0.72-2.85 (46H, m), 4.05-4.48 (3H, m) Elemental analysis C ₃₀ H ₅₆ N ₄ O ₇ Calculated value c: 61.61, H: 9.65, N: 9.58 Measured value c: 61.64, H: 9.52, N: 9.47

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Yoshihiko Kitaura 523 Awaden, Sakurai City, Nara Prefecture (72) Inventor Osamu Nakaguchi 1-5-10 Shiroyamacho, Toyonaka City, Osaka Prefecture (Green Heights Sone 418) (72) Invention Eiji Hemi S-501, South 8 Aobaoka, Suita City, Osaka Prefecture S-501 (72) Inventor Matsuhiko Aratani 1-4 Satakedai, Suita City, Osaka Prefecture A13-108 (72) Inventor Shuichi Takeno 67 Maenashicho, Tenri City, Nara Prefecture (72) Inventor Tatsu Okada 2-21-7 Kawazoe, Takatsuki-shi, Osaka (72) Inventor Hirokazu Tanaka 3-10-21 Hanayashiki Manor, Takarazuka-shi, Hyogo Prefecture (72) Masashi Hashimoto 1-6 Nakagayadai, Takarazuka-shi, Hyogo Prefecture -17 (56) Reference JP-A-57-114556 (JP, A) JP-A-56-45449 (JP, A)

Claims (1)

[Claims] 1. A general formula Wherein R ¹ represents palmitoyl, R ² represents hydrogen or benzyl, and R ₃ represents hydrogen or benzyloxycarbonyl, respectively, and a pharmaceutically acceptable salt thereof.