JPH0325437B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention relates to a novel peptide compound, and in particular to a novel peptide compound having pharmacological activity or a pharmaceutically acceptable salt thereof, a method for producing the compound, and a pharmaceutical composition containing the compound. The present inventors have so far succeeded in synthesizing many peptide compounds and have completed patent applications in many countries (for example, Japanese Patent Application No. 106279/1982). Subsequently, as a result of further research, the present inventors succeeded in synthesizing new compounds, which are not specifically disclosed in the above application specification, and found that these compounds had excellent preventive effects and anti-inflammatory effects on experimental infectious diseases. They discovered that it has a cancer effect and completed this invention. The novel peptide compound of the present invention is represented by the following formula (). (In the formula, R ¹ means n-octanoyl or stearoyl, R ² means hydrogen or benzyl, R ³ means 1-carboxyethylamino, R ⁴ means hydrogen, and R ⁵ means hydrogen or benzyloxycarbonyl.) General formula () Pharmaceutically acceptable salts of the novel peptide compound represented by include, for example, alkali metal salts (e.g., sodium salts, potassium salts, etc.), alkaline earth metal salts (e.g., calcium salts, etc.), ammonium salts, and organic amine salts. (e.g., ethanolamine salt, triethylamine salt, dicyclohexylamine salt, etc.), with organic or inorganic acids such as methanesulfonate, hydrochloride, sulfate, nitrate, phosphate, etc. Examples include acid addition salts of. The novel peptide compounds () of the present invention can be synthesized by various methods detailed below. () Peptide bond formation reaction: () Protecting group elimination reaction: (In the formula, R ¹ means n-octanoyl or stearoyl, and R ³ means 1-carboxyethylamino.) [] Peptide bond formation reaction [(1) to (2)]: This reaction is performed as follows. Can be done. That is, first, the carboxy group of each carboxy compound or its salt is converted into, for example, an acid halide, an acid azide, an acid anhydride, or a mixed acid anhydride, by a conventional method.
The target compound can be synthesized by converting it into an activated form such as an active ester and then reacting it with each amino compound, and also by combining each carboxy compound or its salt with each amino compound or its salt using a commonly used condensing agent. For example, the target compound can also be synthesized by direct reaction in the presence of N,N-dicyclohexylcarbodiimide or the like. Among the above activation methods, a suitable activation method and a suitable condensing agent are appropriately selected depending on the carboxy protecting group of each carboxy compound, the type of each amino compound, and reaction conditions (e.g. reaction solvent, temperature, etc.) . This reaction is carried out in a solvent such as methylene chloride, chloroform, tetrahydrofuran, dioxane, ethyl acetate, methanol, ethanol, or water.
Proceeds smoothly at temperatures ranging from -20°C to room temperature. The reaction in the presence of a condensing agent is usually carried out under mild conditions under anhydrous conditions. [] Protecting group elimination reaction: This reaction is a method for synthesizing compound ( ^b ) or its salt by subjecting compound ( ^a ) or its salt to amino and/or carboxy protecting group elimination reaction. It is. [-1] Elimination reaction of amino protecting group: This reaction can be carried out by conventional methods such as reduction methods such as catalytic reduction, liquid ammonia alkali metal method, method using acid, zinc acid method, method using base, hydrazine method, etc. It is carried out by. The reaction temperature is usually about cooling to room temperature. [-2] Carboxy protecting group elimination reaction: This reaction is carried out by a conventional method such as hydrolysis or reduction. Hydrolysis normally proceeds smoothly in a solvent, in the presence of an acid or a base, and under relatively mild conditions such as cooling or heating. Reduction includes chemical reduction and catalytic reduction, and is carried out according to conventional methods.
The reaction is usually carried out in a solvent under relatively mild conditions such as cooling or heating. The starting compounds used in this invention include known compounds (for example, European Patent Publication No. 11283) and new compounds, and the new compounds can be synthesized by the same method as described above. The object compound [] and the starting compound of this invention contain one or more isomers due to asymmetric carbon atoms in the molecule, and all such isomers are also included in the scope of the invention. It has been revealed that the novel peptide compound () and its pharmaceutically acceptable salts provided by the present invention have preventive effects against experimental infectious diseases and anticancer effects. Therefore, the novel peptide compound () and its pharmaceutically acceptable salts are useful substances for the treatment of infectious diseases caused by pathogenic microorganisms, especially gram-negative bacteria, gram-positive bacteria, and fungi, and for the treatment of cancer in humans and various animals. . Pharmacological data will be explained below to demonstrate the usefulness of the novel peptide () as a medicine. Preventive effect against experimental infectious diseases in mice: For the purpose of determining the preventive effects against experimental infectious diseases in mice, test compounds were dissolved and diluted in physiological saline to prepare test solutions at specified concentrations. 10 male ICR mice (4 weeks old)
grouped. E. coli 22 was cultured on Trypticase Soy agar medium at 37° C. overnight and suspended in physiological saline to obtain a suspension having a microbial cell concentration of 2.6×10 ⁹ CFU/ml. 8.7× per mouse
10 ⁷ CFU of bacterial cells were administered intraperitoneally. In addition, regarding the test compound, for 10 mice in one group,
Various doses were previously administered intraperitoneally for 4 days. The survival rate was determined from the number of surviving animals on the 3rd day after the administration of the bacterial cells.
It is listed in the table.

[Table] Anticancer effect: (1) Methylcholanthrene-induced fibrosarcoma cells (Meth-A) were used. MethA tumor cells were transplanted into the thoracic cavity of BALB/C mice. Test compounds were administered 14 days before MethA implantation;
It was administered intrathoracically three times, 1 hour and 3 days after transplantation. The anticancer effect of the test compound was investigated using survival prolongation of MethA-implanted mice as an indicator. The results are shown in the table below.

[Table] The pharmaceutical composition of the present invention can be used in various pharmaceutical formulations,
For example, the active substance of the invention may be provided as a solid, semi-solid or liquid formulation, containing an organic or inorganic carrier or excipient suitable for external, internal or topical application. The active ingredient may be present in tablets, pellets, capsules, suppositories, solutions, emulsions,
It is used in combination with non-hazardous and pharmaceutically acceptable auxiliary ingredients to form a suspension or other suitable form. Such auxiliary ingredients include ingredients effectively used in the production of solid preparations, semi-solid preparations, liquid preparations, etc., such as water, glucose, lactose, gelatin, mannitol,
Examples include starch paste, magnesium trisilicate, corn starch, keratin, colloidal silica, potato starch, urea, etc., and stabilizers, fillers, colorants, fragrances, etc. may also be added as supplements.
Furthermore, the pharmaceutical composition of the present invention may contain a preservative or a bactericidal agent in order to maintain the activity of the active ingredient. The amount of the active ingredient in the composition is such that the amount of the active ingredient is sufficient to exert a desired therapeutic effect on the progress and condition of the disease. When applying this composition to the human body, intravenous administration, intramuscular administration, oral administration, etc. are desirable.
Although the effective dose of the substance of the present invention varies depending on the age of the target patient and the symptoms, the usual daily dose for humans or animals is 0.1 to 100 mg per 1 kg of body weight.
The content in the formulation is approximately 50mg, 100mg, 250mg, 500mg
It is common to do so. Examples of the present invention will be described below. In the following production examples and examples, starting materials and target materials are represented using the following abbreviations. Ala: Alanyl Glu: Glutamine DAP: α,ε-diaminopimelyl Z: Benzyloxycarbonyl Boc: t-butoxycarbonyl Bzl: Benzyl Su: N-hydroxysuccinimide Lys: Lysyl production example D-AlaOH (2) (1.78 g) was dissolved in a mixed solvent of water (40 ml), dioxane (40 ml), and triethylamine (4.04 g). Add Boc-L-Lys (ε
-Z)OSu(1) (9.26 g) was added and the resulting solution was reacted overnight at room temperature, then filtered. The oily substance obtained by evaporating the filtrate was dissolved, and the solution was acidified by adding dilute hydrochloric acid, and then extracted with ethyl acetate. After washing the organic solvent layer with water and drying with magnesium sulfate, the solvent was distilled off to obtain a white foam. The foam was dissolved in trifluoroacetic acid (30ml) and allowed to react for 30 minutes at room temperature. Excess trifluoroacetic acid was distilled off and the resulting paste was dissolved in water. After passing the solution through an HP20 column, elution was performed by passing water and water-methanol (1:1) through the column for + minutes. The latter eluate was collected and the solvent was distilled off to give L-Lys(ε-Z)-D-AlaOH(3) (4.50g)
was gotten. IR (Nujol): 3350, 3300, 1685, 1660, 1640cm ^-1 NMR (CD ₃ OD): δ1.30 (3H, d, J = 7Hz),
1.20−1.70 (6H, m), 2.86−3.50 (2H, m),
4.16 (1H, q, J = 7Hz), 5.00 (2H, s),
7.30 (5H, s) Example 1 L-Lys(ε-Z)-D-AlaOH(2) (0.915g)
was dissolved in a mixture of methylene chloride (40 ml), methanol (40 ml) and triethylamine (0.53 g). Add n-octanoyl-D-Glu (α
-OBZl)-γ-OSu(1) (1.20 g) was added and left at room temperature overnight. The solvent was distilled off from the reaction mixture, and water (50 ml), ether (50 ml), and IN hydrochloric acid were added to the paste-like residue. The resulting mixed solution was sufficiently stirred and the ether was distilled off, and crystals were precipitated from the remaining aqueous layer. The crystals are filtered, washed with water, and dried, resulting in n-octanoyl-γ-D-Glu (α
−OBZl)−L−Lys(ε−Z)−D−AlaOH(3)
(1.50g) was obtained. IR (Nujol): 3300, 1725, 1685, 1650, 1630 cm ^-1 NMR (DMSO-d ₆ ,) δ: 0.84 (3H, t, J = 7
Hz), 1.00-2.40 (25H, m), 2.84-3.12 (2H,
m), 4.10−4.50 Octanoyl-γ-D-Glu(α-OBZl)-L
-Lys(ε-Z)-D-AlaOH (3) (1.20 g) was dissolved in acetic acid (50 ml) and hydrogenated using palladium black (150 mg). The filtrate from which the catalyst has been filtered off is evaporated,
When the residual paste was allowed to stand, crystals were obtained. When the crystals were thoroughly washed with diethyl ether, octanoyl-γ-D-Glu(α-OH)-L-Lys-D
-AlaOH(4) (0.80 g) is obtained. [α] _D = +41.7 (C = 0.2, acetic acid) IR (Nujol): 3360, 1710 (sh), 1640 cm ^-1 NMR (D ₂ O), δ: 0.84 (3H, obscure t, J =
7Hz), 1.00-2.50 (25H, m), 2.80-3.10
(2H, m), 4.00−4.40 (3H, m) Example 2 In the same manner as step (1) of Example 1, stearoyl-γ-D-Glu(α-OBzl)-L-Lys(ε-Z)-
D-AlaOH (3) was obtained. NMR (DMSO- _d6 ), δ: 0.84 (3H, t, J=7
Hz), 1.00-2.40 (45H, m), 2.80-3.10 (2H,
m), 4.00−4.80 (3H, m), 5.00 (2H, s),
5.08 (2H, s), 7.32 (10H, s), 7.80 (1H,
d, J = 8Hz) 8.08 (2H, t, J = 8Hz) Stearoyl-γ-D-Glu(α-OH)-L-Lys-D-
AlaOH (4) was obtained. [α] _D = -11.10 (C = 0.21, acetic acid) IR (Nujol): 3350, 1730, 1640 cm ^-1 NMR (NaOD - D ₂ O), δ: 0.68 - 2.80 (50H,
m), 4.10−4.50 (3H, m)

Claims (1)

[Claims] 1. General formula (In the formula, R ¹ is n-octanoyl or stearoyl, R ² is hydrogen or benzyl, R ³ is 1-carboxyethylamino, R ⁴ is hydrogen, and R ⁵ is hydrogen or benzyloxycarbonyl). A compound or a pharmaceutically acceptable salt thereof.