JPS6239151B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention is based on the formula The present invention relates to a novel α-aminooxycarboxylic acid derivative and a salt thereof. In the above formula, R ₁ represents an acyl group substituted with an α'-amino group. This acyl group further includes a hydroxyl group, an amino group,
It may be substituted with a carboxyl group or a phenyl group. R ₂ is a hydrogen atom, a lower alkyl group, a phenyl group (this phenyl group may be substituted with a lower alkyl group or a halogen atom), or a group -(CH ₂ ) _o COOH (n is an integer of 1 to 6). ). Examples of the salt of the α-aminooxycarboxylic acid derivative represented by formula () include amine acid salts and alkali and alkaline earth metal salts of carboxylic acids; examples of amine acid salts include hydrochloric acid, Sulfuric acid, nitric acid, oxalic acid, maleic acid,
Pharmaceutical non-toxic salts such as tartaric acid, malonic acid, citric acid, phthalic acid, naphthalene disulfonic acid salts, and alkali and alkaline earth metal salts of carboxylic acids such as lithium, sodium, potassium, magnesium or calcium salts. Examples include pharmaceutical non-toxic salts of The compound of the present invention is an antibacterial substance having activity mainly against Gram-negative bacteria and is useful as a medicine. Representative examples of the compound having the general formula () include the following compounds. The α-aminooxycarboxylic acid derivative represented by the general formula () can be produced by the following reaction formula. (In the formula, X represents a halogen atom, and R ₂ is the same as described above.) An anhydrous ethanol solution of benzohydroxamic acid (1) was added to an ethanol solution of sodium ethoxide made from metallic sodium and anhydrous ethanol, when cold or Add it dropwise while heating to make the sodium salt of (1). If the sodium salt is made completely, it may be heated. After several hours, an anhydrous ethanol solution of α-halogenoacetic acid derivative (2) is added dropwise, and the mixture is allowed to react for several hours to overnight at room temperature or under ethanol reflux. After adding ice in the usual manner, ethanol is distilled off, the mixture is made acidic (pH 3 or less) with hydrochloric acid or sulfuric acid, and extracted with ethyl acetate or sometimes butanol to obtain an acidic substance. A pure product (3) can be obtained from the obtained acidic substance by chromatography using silica gel. Next, (3) is hydrolyzed with a 1:1 mixture of hydrochloric acid and glacial acetic acid to obtain the hydrochloride of aminooxycarboxylic acid (4) in which the benzoic acid moiety has been hydrolyzed. (In the formula, Y is from the α′-amino group-substituted acyl group defined in R ₁ above.

Residues excluding [Formula] are shown. Z represents a protecting group for an amino group, and is usually a carbobenzyloxy group. Further, Suc represents a succinimide group. ) (4) is made alkaline (pH 9 or higher) using triethylamine in anhydrous dimethylformamide, and the solution is mixed with a neutral amino acid with a protected amino group, an active ester activated with the succinimide group of an aromatic amino acid, or an amino acid. By adding an active ester of succinimide of a basic amino acid with a protected group, or an active ester of succinimide of an acidic amino acid with a protected amino group and a carboxyl group that does not participate in the reaction with a t-butyl group, etc. The resulting compound (6) is purified by chromatography using silica gel, etc., and the protecting group of the amino group or carboxyl group is specifically removed by reduction or acid hydrolysis to obtain compound (7). . The α-aminooxycarboxylic acid derivatives and salts thereof of the present invention are antibacterial substances that are active mainly against Gram-negative bacteria. The antibacterial activity of some of these compounds against Escherichia coli NIHJ JC-2 strain was tested using the disc method, and the results shown in Table 1 were obtained.

[Table] These compounds can also be combined with known antibacterial substances to form compositions with a wide range of antibacterial activity. Examples and reference examples are shown below. Reference Example 1 60 ml of an anhydrous ethanol solution of 2.7 g of benzohydroxamic acid is added dropwise at 25°C to 30 ml of an anhydrous ethanol solution of sodium ethoxide prepared by dissolving 0.92 g of sodium metal in 30 ml of anhydrous ethanol. Stir well for 3 hours. A solution of 1.96 g of α-bromosuccinic acid in 10 ml of absolute ethanol is added dropwise, and the mixture is kept stirring at 50° C. for 8 hours. After cooling, water was added, neutralized with hydrochloric acid, ethanol was distilled off, and the mixture was further diluted with hydrochloric acid and extracted with ethyl acetate. Column chromatography was carried out using chloroform:ethyl acetate (1:1) to obtain α-(N-benzyl)aminooxysuccinic acid (1 g). IR (ν ^KBr _cn-1 ): 3450, 33250, 2650, 2500,
1740, 1720, 1620, 1580, 1280, 690 RMR (δ _ppn , _(CD3)2CO ): 7.5-8.0 (5H, m)
4.95 (H, q), 3.0 (2H, dd) Reference Example 2 150 mg of α-(N-benzyl)aminooxysuccinic acid obtained in Reference Example 1 was mixed with concentrated hydrochloric acid:glacial acetic acid (1:1).
The mixture was stirred overnight at room temperature, the resulting benzoic acid was removed by extraction with ethyl acetate, the aqueous layer was concentrated under reduced pressure, and finally freeze-dried to obtain 84 mg of aminooxysuccinic acid powder. . Cellulose thin layer chromatography showed antibacterial activity against Escherichia coli NIHJ JC-2 strain at R _f 0.2 using n-butanol/acetic acid/water (upper layer of 4:1:5) as a developing solvent. PMR (δ _ppn , _D2O ): (PMR measured in heavy water
is based on water 4.8. Same below. )5.0
(H, t), 3.0 (2H, d) Example 1 (Production method of Exemplary Compound No. 13) 234 mg of aminooxyacetic acid hydrochloride was dissolved in anhydrous dimethylformamide, triethylamine was added dropwise to make it alkaline, and α-amino group Active ester of succinimide of valine protected with carbobenzyloxy group [GWAnderson et al.
It was synthesized by the method of JACS 86 , 1839 (1964). ] Add 400 mg and stir at room temperature overnight.
The excess active ester is (CH ₃ ) ₂ NCH ₂ CH ₂ CH ₂ NH ₂
The mixture was decomposed by adding and acidified with hydrochloric acid, and then extracted with ethyl acetate. The ethyl acetate layer is washed with water, dried over Na ₂ SO ₄ and the solvent is distilled off. Next, the residue was dissolved in a mixture of methanol and water (7:3), 120 mg of 10% Pd-C was added, and hydrogen was passed through the solution for reduction. Pd after reaction
-C was removed by filtration, neutralized with NH ₄ OH water, the solvent was concentrated, and finally freeze-dried to obtain a powder. This powder was subjected to column chromatography using Sephadex G-15 (200 ml), developed using the upper layer of n-butanol:acetic acid:water (4:1:5), and the portion that was positive with ninhydrin was collected. ,
Concentration yielded 140 mg of a single powder. This product showed antibacterial activity at R _f 0.25 in the same thin layer chromatography as in Example 2. IR (ν ^KBr _cn-1 ): 3450, 1690, 1600, 1400, 13
20,
1070 PMR (δ _ppn , _D2O ): 4.40 (2H, s) 3.65 (H,
d), 2.3 (H, m), 1.1 (3H, d), 1.08
(3H, d) Example 2 (Production method of Exemplary Compound No. 14) 130 mg of aminooxyacetic acid hydrochloride was dissolved in 4 ml of anhydrous dimethylformamide, made alkaline by adding triethylamine, and protected the amino group in the same manner as in Example 1. A reaction was carried out by adding 230 mg of active ester of leucine, followed by post-treatment in the same manner, and finally freeze-drying to obtain 74 mg of powder. This substance showed antibacterial activity at R _f 0.35 in thin layer chromatography similar to Reference Example 2. PMR (δ _ppn , _D2O ): 4.42 (2H, s), 4.12
(1H, t), 2.02 (H, m), 1.85 (2H, m),
1.1 (6H, d) Example 3 (Production of Exemplary Compound No. 15) 100 mg of aminooxyacetic acid hydrochloride was dissolved in 3 ml of dimethylformamide, triethylamine was added to make it alkaline, and the amino group was protected in the same manner as in Example 1. Active ester of phenylalanine 200
mg was added to carry out the reaction, followed by post-treatment in the same manner and finally freeze-drying to obtain 50 mg of powder. This material showed antibacterial activity with R _f 0.28 in thin layer chromatography similar to Example 2. PMR (δ _ppn , _D2O ): 7.4 (5H, s), 4.75 (peak overlaps with water), 3.8 (H,
t), 3.05 (2H, dd) Example 4 (Production of Exemplary Compound No. 16) Dissolve 100 mg of aminooxyacetic acid hydrochloride in 3 ml of dimethylformamide, add triethylamine to make it alkaline, and convert the ε-amino group to t. Example 1: 200 mg of activated ester of lysine with -butyl and α-amino groups protected with carbobenzyloxy groups.
reacts in the same manner as , and the carbobenzyloxy group is 10%
The t-butyl group was removed by treatment with trifluoroacetic acid, and partition chromatography was performed using Sephadex G-15 in the same manner as in Example 1 to obtain the active substance. In cellulose thin layer chromatography using 70% n-propanol as a developing solvent, antibacterial activity was found at the origin against Escherichia coli NIHJ JC-2 strain. Example 5 (Production method of Exemplary Compound No. 9) Hydrochloride of α-aminooxy-α-methylacetic acid
Dissolve 100mg in 3ml of dimethylformamide, add triethylamine dropwise to make it alkaline, and prepare active ester of leucine with amino group protected.
Add 230mg and react. Post-treatment was carried out in the same manner as in Example 1 to obtain 105 mg of coarse powder. Thin layer chromatography using cellulose and developing with an upper layer of n-butanol:acetic acid:water (4:1:5) gave a single spot at R _f 0.35 with ninhydrin coloring. Antibacterial activity was also found in the same spot. Example 6 (Production method of Exemplary Compound No. 7) Hydrochloride of α-aminooxy-α-methylacetic acid 70
mg was dissolved in 3 ml of dimethylformamide, made alkaline by adding triethylamine, and added with 200 mg of an active ester of glutamic acid in which the α-amino group was protected with a carbobenzyloxy group and the γ-carboxyl group was protected with a t-butyl group. React in the same way. Post-treatment was carried out in the same way, making it hydrochloric and acidic.
Perform ethyl acetate extraction, evaporate the ethyl acetate layer, dissolve the residue in methanol, and add 10% Pd-C100.
After filtration, the solution is concentrated, and the residue is dissolved in trifluoroacetic acid, and reacted at room temperature for 15 minutes to remove the t-butyl group. Trifluoroacetic acid was distilled off under reduced pressure, and the residue was transferred to Cephadex G-
Partition chromatography was performed using 15 to obtain a single substance, ninhydrin, which has antibacterial activity.
In silica gel thin layer chromatography, n
-Propanol:pyridine:acetic acid:water (15:10:
E. coli NIHJ at R _f 0.3 using 3:12) as the developing solvent.
Antibacterial activity against the JC-2 strain was present. PMR (δ _ppn , _D2O ): 4.9 (overlap with water), 4.0 (m, H), 1.9-2.7 (m, 4H), 1.45
(d, 3H) Example 7 (Production method of Exemplary Compound No. 40) Hydrochloride of aminooxysuccinic acid obtained in Example 2
50 mg of Pd-C was dissolved in 5 ml of dimethylformamide, made alkaline with triethylamine, added with 80 mg of active ester of valine whose α-amino group was protected with a carbobenzyloxy group, and reacted in the same manner as in Example 1. ) to remove the carbobenzyloxy group, partition chromatography was performed using Sephadex G-15, and finally freeze-drying to obtain 26 mg of a single powder.
This material showed antibacterial activity at R _f 0.15 in thin layer chromatography similar to Example 2. IR (ν ^KBr _cn-1 ): 3350, 1690, 1600, 1390 PMR (δ _ppn , _DMSO ) 4.2 (H, q), 3.1 (H,
d), 2.6 (2H, overlap with DMSO),
1.90 (H, m), 0.85 (6H, d) Example 8 (Production method of Exemplary Compound No. 41) Dissolve 50 mg of aminooxysuccinic acid hydrochloride in dimethylformamide, add triethylamine dropwise to make it alkaline, The reaction was carried out in the same manner as in Example 1 by adding 80 mg of active ester of group-protected leucine, and the same post-treatment was carried out to collect a single fraction with ninhydrin, which has antibacterial activity.Finally, it was freeze-dried to obtain a crude product. 50 mg of powder was obtained. In silica gel thin layer chromatography, this product was
Using the upper layer of butanol:acetic acid:water (4:1:5) as a developing solvent, it exhibited antibacterial activity against Escherichia coli NIHJ JC-2 strain at R _f 0.1.

Claims (1)

[Claims] 1 formula (In the formula, R ₁ represents an α′-amino group-substituted acyl group (this acyl group may be further substituted with a hydroxyl group, an amino group, a carboxyl group, or a phenyl group), and R ₂ represents a hydrogen atom, a lower an alkyl group, a phenyl group (which may be substituted with a lower alkyl group or a halogen atom), or a group-
(CH ₂ ) _o COOH (n represents an integer from 1 to 6). ) α-Aminooxycarboxylic acid derivatives and salts thereof.