JPH0372205B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention is based on the formula (wherein, X represents a halogen atom, R ₁ represents a hydrogen atom or a lower alkyl group, and R ₂ represents a lower alkyl group) or a salt thereof as an active ingredient for mycoplasma of animals (excluding humans) Concerning preventive and therapeutic agents for infectious diseases. Mycoplasma is a type of bacteria that lacks a cell wall and is the smallest of all bacteria, and many subspecies are known to infect humans and animals. BACKGROUND ART In recent years, the number of livestock and poultry raised per unit area has increased, and large-scale livestock farming has developed, resulting in the occurrence of various infectious diseases, and the damage caused by them is increasing year by year. In particular, mycoplasma causes single infections or mixed infections with common bacteria such as E. coli. Such infections induce intractable respiratory diseases, arthritis, reproductive disorders, etc., resulting in large losses to the livestock industry. Traditionally, tylosin has been most frequently used for the preventive treatment of mycoplasma infections, and has greatly contributed to the development of the livestock industry. However, these antibiotics cannot eliminate mycoplasma in the body;
Moreover, recently, multidrug-resistant strains have appeared, and the original preventive and therapeutic effects can no longer be expected. Therefore, the present inventors completed the present invention as a result of intensive studies to find a compound that has an excellent effect on mycoplasma infection in animals. That is, the present invention provides a method for preventing and treating mycoplasma infection in animals containing the compound of formula (1) or a salt thereof as an active ingredient.
It relates to therapeutic agents. Examples of animals include various beasts such as livestock such as cows and pigs, and poultry such as chickens, and birds. Mycoplasma infections include infections caused by mycoplasma alone and mycoplasma and other infections.
This can include bacterial infections of different species or mixed infections with two or more types of bacteria. Examples of mycoplasma-only infections include mycoplasma pneumonia, mycoplasma mastitis, and mycoplasma arthritis in cattle, epidemic pneumonia and mycoplasma arthritis in pigs, and chronic respiratory disease and infectious arthritis in chickens. It will be done. Examples of mixed infections include Mycoplasma and one or more of E. coli, S. aureus, Str. pneumoniae, H. influenzae, etc. Pneumonia or mastitis in cattle caused by bacteria, Mycoplasma and Escherichia coli, P.multocid, H.parahemolyticus, B.bronchiseptila, etc., or one or more of the following bacteria. These include epidemic pneumonia in pigs caused by mycoplasma and chronic respiratory disease in chickens caused by one or more bacteria such as Escherichia coli, staphylococcus, and H.paragllinarum. Next, an in vitro antibacterial activity test of the compound of formula (1) against mycoplasma and an experimental prophylactic/therapeutic test against mycoplasma monoinfection and mixed infection will be shown. Experimental Example 1 Antibacterial activity test against mycoplasma derived from livestock and poultry in vitro This test was conducted using 9 strains of mycoplasma derived from cattle, pigs, and chickens using a liquid medium or agar medium dilution method. 9-Fluoro-10-(4-methyl-1-piperazinyl)-7-oxo-2,3-dihydro-7H-pyrido[1,2,3-de]-1,4
-Benzoxazine-6-carboxylic acid (compound A) and 9-chloro-10-(4-methyl-1-piperazinyl)-7-oxo-2,3-dihydro-
Table 1 shows the results of measuring the antibacterial activity of 7H-pyrido[1,2,3-de]-1,4-benzoxazine-6-carboxylic acid (compound (B)) and tylosin used as a control against mycoplasma. Also, 3S−
9-Fluoro-2,3-dihydro-3-methyl-
Antibacterial activity of 10-(4-ethyl-1-piperazinyl)-7-oxo-7H-pyrido[1,2,3-de][1,4]benzoxazine-6-carboxylic acid (compound C) against mycoplasma in chickens The force measurement results are shown in Table 2.

【table】

【table】

[Table] Medium 1 100ml medium composition PPLO medium (manufactured by Eiken) 1.8g Yeast extract 0.5g L-glutamine 0.025g Arginine hydrochloride 0.025g 0.2% DNA (deoxyribonucleic acid, manufactured by Sigma)
1.0ml 1.0% β-NAD (β-nicotinamide adenine dinucleotide, manufactured by Sigma) 1.0ml 1.0% phenolic acid 0.2ml Water 80.0ml Pig serum (manufactured by Irvin) 20.0ml Medium 2 2 g of agar is added to the composition of medium 1. plus. Medium 3 100ml medium composition PPLO medium 3.5g 0.2% DNA 1.0ml 1.0% DPN (diphosphopyridine nucleotide,
(manufactured by Sigma) 1.0ml 1.0% phenol red 0.2ml Water 80.0ml Horse serum (manufactured by Nippon Bio) 20.0ml Medium 4 A version of medium 3 in which horse serum was replaced with pig serum. Experimental Example 2 Preventive test of Compound A, Compound B, and Tylosin against mycoplasma monoinfection Ten groups of 10 chicken chicks (10 days old) were set up. Of these, 6 groups were treated with Compound A and Compound B, and 2 groups were treated with Tylosin. The remaining two groups were an infected control group and a non-infected control group. Compound A and compound B were uniformly 50 and 75, respectively.
and powdered feed (standard feed for chicken testing SDL.No.1, manufactured by Japan Compound Feed Co., Ltd.) to a concentration of 100 ppm.
The mixture was mixed with the following drugs and administered at free temperature for 5 days starting 3 days before infection. Tylosin was dissolved in drinking water at concentrations of 250 and 500 pm and administered in the same manner. The infected control group and the non-infected control group were fed powdered feed and drinking water without any drug added. Infection is Mycoplasma gallisepticum
0.2 ml (7.2×10 ⁷ chicks/chicken) of a 48-hour culture of strain 934N (hereinafter Mg) (medium 3 of Experimental Example 1) was inoculated into the trachea of chicks in each group except the non-infected control group. Judgment was made as follows. That is, after the completion of the administration, the animals were reared with drug-free feed and drinking water for an additional 5 days, and then sacrificed by exsanguination.The presence or absence of lesions characteristic of mycoplasma infection was observed through autopsy, and bacteria were detected from the air sacs and trachea. The test results are shown in Table 3. As shown in Table 3, tylosin was unable to prevent Mg-only infection even at a dosage of 500 ppm, whereas compound A was unable to prevent Mg infection at a concentration of 100 ppm and compound B was 75 ppm.
Complete prevention was confirmed at high dosages.

[Table] Example 3 Treatment test of compound A, compound B, and tylosin against mycoplasma monoinfection A 48-hour culture of Mg bacteria (medium 3 of Experimental Example 1) was injected into the trachea of 90 chicken chicks (10 days old). 0.2ml (6.7× ¹⁰⁷
per bird) was inoculated. The animals were divided into 9 groups with 10 birds per group, of which 6 groups were treated with compound A and compound B, 2 groups were treated with tylosin, and 1 group was used as an infection control group. In addition, 10 healthy normal chicks were set as a non-infected control group. Compound A, compound B and tylosin are in Experimental Example 2
The drug was added to powdered feed and drinking water in the same manner as above, and administered ad libitum for 5 days from the day of infection. The infected control group and the non-infected control group were given free access to drug-free feed and drinking water throughout the test period. The determination was made in the same manner as in Experimental Example 2. The test results are shown in Table 4. As shown in Table 4, it was confirmed that compound A can completely treat Mg-only infection at a dosage of 100 ppm and compound B at a dosage of 75 ppm or more, far exceeding the efficacy of tylosin.

[Table] The method for calculating the treatment rate was in accordance with Experimental Example 2.
Experimental Example 4 Prevention test of compound A, compound B, and tylosin against mixed infection of mycoplasma and E. coli Eleven groups of 10 chickens (10 days old) were set up. Of these, 8 groups were administered compound A and compound B mixed in powdered feed at concentrations of 50, 75, 100, and 150 ppm, respectively, and allowed to take them ad libitum for 5 days starting 3 days before infection. The other group was given tylosin as feed.
Mixed to a concentration of 550 ppm and dosed in a similar manner.
Two groups were designated as an infected control group and a non-infected control group, and were fed drug-free feed and drinking water throughout the test period. Infection was carried out using Mg's 48-hour culture medium (medium 3 of Experimental Example 1) and Escherichia coli S6 strain (hereinafter referred to as Ec).
Mix equal amounts of 24-hour culture solution (Trypticase Soy Broth Medium, manufactured by Eiken), and add 0.4 ml (M.
9.4×10 ⁶ E.c. and 1.3×10 ⁷ E.c./chicken) were inoculated into the airways of chicks in each group except the non-infected control group. Judgment was made based on the number of deaths during the test period after feeding the animals with drug-free feed and drinking water for an additional 5 days after the end of the administration, and the autopsy and bacterial tests at the end of the test. The test results are shown in Table 5. As shown in Table 5, compound A and compound B are
It was confirmed that mixed infection of Mg and Ec was completely prevented at a dosage of 100 ppm or more, and its preventive effect was far superior to that of tylosin.

【table】

[Table] Experimental Example 5 Treatment test of Compound A, Compound B, and Tylosin against a mixed infection of Mycoplasma and Escherichia coli Experimental Example 4 in the air of 200 chicken chicks (10 days old)
0.4ml of mixed bacterial solution (M.g4.7×10 ⁶ ,
E.c6.6×10 ⁷ birds/wing) were inoculated and mixed infection was carried out. The animals were divided into 10 groups of 20 birds per group, of which 8 groups were treated with compound A and compound B, 1 group was treated with tylosin, and the other group was an infected control group. Separately, 20 healthy normal chicks were set as a non-infected control group. Compound A, compound B and tylosin were used in Experimental Example 4.
It was mixed with powdered feed in the same manner as above and administered for 5 days from the day of infection. The infected control group and non-infected control group were bred in the same way as in other tests. The determination was made in the same manner as in Experimental Example 4. The results are shown in Table 6. As shown in Table 6, compound A
and compound B at a dosage of 150 ppm concentration of Mg and
It was confirmed that Ec mixed infection was completely treated and its therapeutic efficacy was far superior to that of tylosin.

【table】

[Table] The treatment rate was calculated according to Experimental Example 4.
Bacterial recovery rate was determined for surviving cases.
As mentioned above, although the compound of formula (1) is slightly inferior to tylosin in in vitro antibacterial activity, it has very excellent prophylactic and therapeutic effects against single and mixed mycoplasma infections in vivo. This was confirmed. The compound of formula (1) is a known compound described in JP-A-57-46986 and JP-A-58-52290, and its toxicity values are shown in Table 7.

[Table] The dosage of the compound of formula (1) varies depending on the administration method and the species of animal to be administered, but is usually in the range of 5 to 30 mg/Kg per day. The compound of formula (1) is usually added to feed or drinking water;
It is administered orally as a powder or an aqueous solution, but in some cases it can also be administered by injection.
When adding the compound of formula (1) to feed, the concentration is usually
The concentration is usually in the range of 100 to 200 mg/Kg, and when added to drinking water, 50 to 100 ml/1. Further, the compound of formula (1) can be prepared into appropriate preparations, aqueous solutions, syrups and injections by techniques commonly used in this field. Next, a prescription example of the aqueous solvent is described below. Compound A, Compound B or Compound C 5g Acetic acid 26ml Purified water Appropriate amount Total 100ml

Claims (1)

[Claims] 1 formula (wherein, X represents a halogen atom, R ₁ represents a hydrogen atom or a lower alkyl group, and R ₂ represents a lower alkyl group) or a salt thereof as an active ingredient for the prevention of mycoplasma infection in animals. therapeutic agent. 2. The preventive/therapeutic agent according to claim 1, wherein the mycoplasma infection in animals is bovine mycoplasma pneumonia. 3. The prophylactic/therapeutic agent according to claim 1, wherein the mycoplasma infection of animals is epidemic pneumonia of pigs. 4. The prophylactic/therapeutic agent according to claim 1, wherein the mycoplasma infection of animals is chronic respiratory disease of chickens.