JPS634526B2 - - Google Patents

Description

[Detailed description of the invention]

This invention relates to a new antibacterial agent,
In detail, the general formula (In the formula, R is a lower alkanoyl group and n is 2-
The present invention relates to an antibacterial agent containing a phosphonic acid derivative or a pharmaceutically acceptable salt thereof and an aminoglycoside antibiotic or a pharmaceutically acceptable salt thereof. Until now, many antibiotics have been used to prevent and treat infections caused by pathogenic bacteria, but each antibiotic has its own unique antibacterial spectrum, and these antibiotics alone have no antibacterial activity or are ineffective. It is generally known that there are pathogenic bacteria that exhibit only weak antibacterial activity. Therefore,
Regarding the prevention and treatment of infections caused by such pathogenic bacteria, there has been a problem in that antibiotics alone are ineffective or have insufficient effects. After various studies aimed at improving these antibiotics, the inventors discovered that aminoglycoside antibiotics or their salts and phosphonic acid derivatives () or their salts have synergistic antibacterial effects against various pathogenic bacteria. In other words, the combination of aminoglycoside antibiotics or their salts with phosphonic acid derivatives () or They obtained new knowledge that by using these salts in combination, it is possible to enhance antibacterial activity that is sufficiently effective for preventing and treating infectious diseases, and as a result of further intensive research, they completed this invention. The antibacterial agent of this invention contains a phosphonic acid derivative () or a pharmaceutically acceptable salt thereof and an aminoglycoside antibiotic or a pharmaceutically acceptable salt thereof. The lower alkanoyl group in R of the phosphonic acid derivative () used in this invention has 1 to 6 carbon atoms.
means an alkanoyl group, preferred examples of which include formyl and acetyl. Moreover, among the integers in n in the general formula (), 3 is preferable. That is, the most suitable phosphonic acid derivative for use in the present invention is a compound represented by the following formula ('). (In the formula, R' means formyl or acetyl.) Phosphonic acid derivatives () are, for example, German Patent Publication No. 2733658 (Publication date: February 9, 1978)
It is a known substance described in the publication and can be produced by the organic synthesis method described in the publication, and the most suitable phosphonic acid derivative (') is Streptomyces ruberomulinas.
3-(N-acetyl-N-hydroxyamino)propylphosphonic acid or 3-(N-formyl-N-hydroxyamino) was obtained by culturing Streptomyces labendulae ATCC31279 in a medium. It can also be produced by obtaining propylphosphonic acid. Further, examples of the aminoglycoside antibiotics used in the present invention include known antibiotics such as gentamicin, tobramycin, dibekacin, amikacin, and bekanamycin. Pharmaceutically acceptable salts of the above phosphonic acid derivatives () include, for example, metal salts such as sodium salt, potassium salt, calcium salt, barium salt, magnesium salt, ammonium salt, ethanolamine salt, triethylamine salt, procaine salt,
Examples include amine salts such as dibenzylamine salts and dicyclohexylamine salts. Furthermore, examples of pharmaceutically acceptable salts of aminoglycoside antibiotics include acid addition salts such as sulfates. The antibacterial agent of this invention can be used for humans, poultry such as chickens, turkeys, ducks, quail, cattle, etc.
It is useful for treating or preventing infectious diseases caused by pathogenic bacteria in livestock such as horses, pigs, sheep, goats, and mink, and pets such as dogs, canaries, and parakeets. In the antibacterial agent of this invention, the mixing ratio of the phosphonic acid derivative () or its pharmaceutically acceptable salt and the aminoglycoside antibiotic or its pharmaceutically acceptable salt varies depending on the type of pathogenic bacteria, the symptoms of the patient, etc., but in general, 1:
1 to 50:1 (weight ratio), preferably 1:1 to 20:1
(weight ratio), but a normal mixing ratio of 4:1 (weight ratio) is sufficient. The antibacterial agent of this invention is usually prepared in the form of a preparation by combining a phosphonic acid derivative () or a pharmaceutically acceptable salt thereof and an aminoglycoside antibiotic or a pharmaceutically acceptable salt thereof with various pharmaceutically acceptable carriers. administered to humans or animals. Injections or ointments are generally preferred as formulations for administration to humans, and injections or infusions are preferred as formulations for administration to animals. The pharmaceutically acceptable carriers mentioned above generally include solids and liquids commonly used in injections, infusions, and ointments; specific examples thereof include petrolatum, paraffin, and propylene glycol. , polyethylene glycol, carboxymethyl cellulose, magnesium gluconate, sodium citrate, calcium carbonate, calcium hydrogen phosphate,
Examples include aluminum stearate, magnesium stearate, distilled water for injection, sesame oil, peanut oil, and peanut oil. The dosage of the antibacterial agent (bacterial infection prevention and treatment agent) of this invention varies depending on the recipient, symptoms, administration method, etc., but generally, phosphonic acid derivatives () or their pharmaceutically acceptable salts and aminoglycosides are used. A mixture of antibiotics or pharmaceutically acceptable salts thereof is appropriately selected within the range of 0.3 to 20 mg/Kg/day. Furthermore, the antibacterial agent of this invention can be administered in divided doses 2 to 4 times a day, if necessary. The antibacterial agent of this invention has low toxicity, as shown, for example, in the following test example. Test Example 1 (Acute toxicity) (1) Tested mixture (a) Mixture consisting of monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid and gentamicin sulfate salt (4:1 weight ratio) (b) A mixture consisting of the monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid and tobramycin (4:1 weight ratio) (c) 3-(N-formyl-N-hydroxyamino)propyl (d) A mixture consisting of the monosodium salt of phosphonic acid and dibekacin sulfate (4:1 weight ratio). ) (e) A mixture consisting of the monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid and bekanamycin sulfate (4:1 weight ratio) (2) Experimental method 1 of the above test mixtures Physiological saline (PH 6.8) (0.5 ml) containing the following was intravenously injected into three 5-week-old DDY male mice (dose: 50 mg/Kg), and observed for 1 week after administration. (3) Test results No abnormalities were observed in the mice tested when any of the above mixtures was administered. Next, the antibacterial activity and infection prevention effect of the antibacterial agent of this invention against various pathogenic bacteria will be explained using Test Examples 2 to 6. Test Example 2 [Synergistic action of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid and gentamicin] A predetermined amount of monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid, The pathogen was cultured overnight on a nutrient agar medium (manufactured by Difco) containing gentamicin sulfate or a mixture of the monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid and gentamicin sulfate (1:1 weight ratio). Multiple items
Spot inoculated using an inoculator (amount of inoculum: 10 ⁸ cells/ml), and after culturing at 37°C for 20 hours,
MIC was measured. Furthermore, in order to investigate the synergistic effect of antibacterial activity, fractional inhibition concentration was calculated from the obtained MIC values.
The Inhibitory Concentration (hereinafter abbreviated as FIC) value and FIC coefficient were calculated using the following calculation method. Calculation method (a) of the monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid
MIC value: A _O (b) MIC value of gentamicin sulfate: B _O (c) MIC value of monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid and gentamicin sulfate: Cab 3-(N -formyl-N-hydroxyamino)
When the mixture ratio of the monosodium salt of propylphosphonic acid and gentamicin sulfate is m:n (weight ratio), each FIC value and FIC coefficient is calculated by the following equation. Γ Monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid
FIC=m/m+nCab/A _O Γ FIC of gentamicin sulfate=n/m+nCab/B _O Γ FIC coefficient=m/m+nCab/A _O +n/m+nCab/B _O The test results are shown in Table 1 below.

[Synergistic action of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid and gentamicin]

a predetermined amount of the monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid, gentamicin sulfate or a mixture of the monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid and gentamicin sulfate; A nutrient agar medium containing (1:1 and 4:1 weight ratio) was spot-inoculated with a 100-fold dilution of an overnight culture of pathogenic bacteria using a multiple inoculator, and after culturing at 37°C for 18 hours, the MIC The value was measured. The FIC value and FIC coefficient were calculated in the same manner as in Test Example 2. The results are shown in Table 2 below.

[Table] Test example 4 3-(N-formyl-N-hydroxyamino)
Propylphosphonic acid and tobramycin, 3-(N
-formyl-N-hydroxyamino)propylphosphonic acid and dibekacin, 3-(N-formyl-N
The synergistic effects of -hydroxyamino)propylphosphonic acid and amikacin and of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid and bekanamycin were measured in the same manner as in Test Example 3. The results are shown in Tables 3, 4, 5 and 6 below.

【table】

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[Synergistic action of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid and gentamicin at various mixing ratios]

A 10-culture dilution of an overnight culture of pathogenic bacteria was added to a nutrient medium (manufactured by Difco) containing a predetermined amount of monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid and gentamicin sulfate.
0.5% inoculation. After culturing at 37°C for 18 hours, the growth state of the test bacteria was observed. The results are shown in Table 7 below. In the table, the symbol "+" means that the test bacteria grew, and the symbol "-" means that the test bacteria did not grow.

[Table] As is clear from Test Examples 2-5, the antibacterial agent of the present invention exhibits synergistic antibacterial effects against various pathogenic bacteria. Test Example 6 (Infection prevention effect) A predetermined amount (10 ⁷ cells/ml) of pathogenic bacteria suspended in 5% mucin aqueous suspension (0.5 ml) was placed in a cell weighing 23 to 25 g.
ICR-strain male mice (8 mice per group) were inoculated intraperitoneally. After inoculation, the antibiotics listed in the table below were injected subcutaneously, and the survival of the test mice was determined one week after infection to determine the ED ₅₀ value. The results are shown in Table 8 below. Note that the FIC value and FIC coefficient were calculated from the obtained ED ₅₀ value according to the calculation method below. Calculation method (a) of the monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid
ED ₅₀ value: A′ _O (b) ED ₅₀ value of gentamicin sulfate: B′ _O (c) ED ₅₀ of a mixture of monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid and gentamicin sulfate Value: C' _ab 3-(N-formyl-N-hydroxyamino)
When the mixture ratio of the monosodium salt of propylphosphonic acid and gentamicin sulfate is m:n (weight ratio), each FIC value and FIC coefficient is calculated by the following equation. Γ FIC of monosodium salt of 3-(N-formyl-N-hydroxyamino)propylphosphonic acid = m/m + nC' _ab /A' _O Γ FIC of gentamicin sulfate = n/m + nC' _ab /B' _O Γ FIC coefficient =m/m+nC′ _ab /A′ _O +n/m+nC′ _a
b /B′ _O

【table】

[Table] As is clear from the above results, the antibacterial agent of the present invention also exhibits synergistic antibacterial activity in the infection prevention test. Next, examples of this invention will be shown. Experimental example 1 3-(N-formyl-N-hydroxyamino)
Monosodium salt of propylphosphonic acid (80mg)
and gentamicin sulfate (20 mg) into a sterile vial and seal. At the time of use, this mixture is dissolved in sterile distilled water for injection to form an injection. In the same manner as in Example 1, injections of mixtures shown in Examples 2-6 below were produced. Example 2 3-(N-acetyl-
A mixture consisting of the monosodium salt of N-hydroxyamino)propylphosphonic acid (200 mg) and gentamicin sulfate (20 mg) was used. Example 3 3-(N-formyl-
A mixture consisting of the monosodium salt of N-hydroxyamino)propylphosphonic acid (120 mg) and tobramycin (30 mg) was used. Example 4 3-(N-formyl-
A mixture consisting of the monosodium salt of N-hydroxyamino)propylphosphonic acid (200 mg) and dibekacin sulfate (50 mg) was used. Example 5 3-(N-formyl-
A mixture consisting of the monosodium salt of N-hydroxyamino)propylphosphonic acid (200 mg) and amikacin sulfate (100 mg) was used. Example 6 3-(N-formyl-
A mixture consisting of the monosodium salt of N-hydroxyamino)propylphosphonic acid (400 mg) and bekanamycin sulfate (100 mg) was used.

Claims (1)

[Claims] 1. General formula (In the formula, R is a lower alkanoyl group and n is 2-
An antibacterial agent containing a phosphonic acid derivative or a pharmaceutically acceptable salt thereof and an aminoglycoside antibiotic or a pharmaceutically acceptable salt thereof. 2 Phosphonic acid derivative has the general formula (wherein R' means formyl or acetyl) The antibacterial agent according to claim 1, wherein the aminoglycoside antibiotic is gentamicin, tobramycin, dibekacin, amikacin or bekanamycin. 3. Claim 2, wherein the mixing ratio of the phosphonic acid derivative or its pharmaceutically acceptable salt and the aminoglycoside antibiotic or its pharmaceutically acceptable salt is 1:1 to 50:1 (weight ratio) antibacterial agent. 4 The phosphonic acid derivative is 3-(N-formyl-N
3. The antibacterial agent according to claim 2, wherein the aminoglycoside antibiotic is gentamicin. 5 The phosphonic acid derivative is 3-(N-formyl-N
3. The antibacterial agent according to claim 2, wherein the aminoglycoside antibiotic is tobramycin. 6 The phosphonic acid derivative is 3-(N-formyl-N
The antibacterial agent according to claim 2, wherein the aminoglycoside antibiotic is dibekacin. 7 Phosphonic acid derivative is 3-(N-formyl-N
-hydroxyamino)propylphosphonic acid, and the aminoglycoside antibiotic is amikacin. 8 Phosphonic acid derivative is 3-(N-formyl-N
3. The antibacterial agent according to claim 2, wherein the aminoglycoside antibiotic is bekanamycin.