KR101035270B1 - - Composition for Intranasal Spray Comprising Amakasin Its Pharmaceutically Approved Salts and Beta-glucan for Atrophic Rhinitis of Pig - Google Patents

Abstract

The present invention relates to a composition for nasal spraying comprising amikacin and a pharmaceutically acceptable salt thereof and beta-glucan as an active ingredient for the prevention and treatment of swine atrophic rhinitis, and more particularly, absolute of swine atrophic rhinitis. The present invention relates to a composition for spraying the nasal cavity of pigs to eliminate the bacterium Bordetella bronchiseptica and the secondary infectious bacterium Pasteurella multocida D for eradication.

Swine atrophic rhinitis, Bodetella bronchiseptica, Pasteurella multocida type D, intranasal spray

Description

Composition for Intranasal Spray Comprising Amakasin, Its Pharmaceutically Approved Salts and Beta- containing Amikacin and its pharmaceutically acceptable salts and beta-glucan as active ingredients for the prevention and treatment of swine atrophic rhinitis glucan for Atrophic Rhinitis of Pig}

The present invention relates to a nasal spray composition for eliminating Bodetella bronchiseptica and secondary infection bacteria Pasteurella multocida type D bacteria in the nose for the absolute eradication of swine atrophic rhinitis. .

Atrophic Rhinitis (AR) is said to be due to the interaction of Bodetella and Pasteurella. Bronchial sepsis ( Bordetella bronchiceptica ) is a fundamental pathogen, and Pasteurella multocida is also a secondary infection often plays a role in promoting lesions. The Bodetella acts to allow the settlement of Pasteurella by stimulating the nasal mucosal epithelium. Both types of bacteria produce different toxins that cause rhinitis.

The atrophic rhinitis is a disease that occurs in piglets as a chronic wasting disease, causing rhinitis (sneezing), deformation of the nose and facial bones (nose crookedness, snout atrophy, etc.), weight loss, and the like. The disease has been reported to be the highest incidence in the country since it was first reported in 1958 in the US, and caused severe damage to pig farms.

There are two causes of the disease.

The primary cause of bacteria is Bordetella bronchiseptica . The bacteria easily settle and multiply in the nasal mucosa of piglets up to 1-3 weeks of age. However, it does not cause nacrecarpal atrophy in breeding pigs over 8-9 weeks of age.

The second causative bacterium is Pasteurella multocid a type D bacterium. The bacterium causes severe nasal turbinate atrophy not only in young piglets but also in pigs 12 to 16 weeks of age. The toxin produced by Pasteurella multocida is a protein and it is easy to know that the bacterium is a main cause of atrophic rhinitis (AR) because it is a protein and causes only a single injection of the toxin into young pigs. have. However, these bacteria are difficult to settle alone in the nasal mucosa and are easily settled when there is a wound (damage) of the mucosa. Because it can multiply, single infection causes mild symptoms.

In other words, Bodetella bronchiceptica bacteria multiply primarily in the nasal mucosa, causing damage to the nasal turbinates, whereby Pasteurella multocida is settled. It multiplies and causes and deepens the nasal turbinate atrophy by its action.

Atrophic rhinitis is one of the important factors, Bodetella bacteria penetrate and settle the nasal mucosa, damage the nasal mucosa, and the infection of the pasteurella complex intensifies and progresses.

Looking at the pathogenesis of atrophic rhinitis (AR), environmental factors cause damage to the nasal mucosa due to stimulation of ammonia gas and invasion of AR-causing bacteria, resulting in destruction of osteoblasts. Humidity control immaturity and impressionist pigs appear.

Since these problems help pneumonia to easily invade the lungs, various pneumonia symptoms appear. In particular, the incidence of "pandemic pneumonia" is very high, and the occurrence of "pasteurella pneumonia" and "pleural pneumonia" is high. The result is slower rates, higher mortality rates, and higher farm production costs.

Relationship between onset and age Infection date Within 1 week Within 3 weeks 4 to 6 weeks of age After 8 weeks Foot incidence 100% 90% 50 to 60% 0-50% Nacre
Atrophy Complete nasal bone
atrophy
++++ Strong lesions

+++ Weak lesion

+ ?? ++ Almost none

ㅁ,- Clinical symptoms Very severe Severe symptoms Weak symptoms Infection, multiplication, blood
Antibodies, no outbreaks

As can be seen from Table 1 above, it can be seen that the onset of atrophic rhinitis (AR) is correlated with the age of infection.

The younger the age of infection, the more easily the onset and symptoms and lesions become more severe, especially the infection of piglets within 3 weeks of age is the biggest problem. In particular, most infections do not occur, and carriers that excrete AR bacteria are causes of repeated transmission of bacterial infections in farms, and adult pigs and sows also become infectious agents of the disease.

Figure 1 is a schematic diagram showing the nasal cross-section of healthy pigs and pigs with atrophic rhinitis.

Referring to Figure 1, cut the nasal cavity of pigs with atrophic rhinitis nasal mucosa is inflammatory and mucus or purulent exudate is attached. Even if there is no change in appearance, cutting between the first and second prosthesis with a saw can determine whether atrophy of the nasal tendon and curvature of the nasal septum. If the change in the nose is not severe, you can check the atrophy of the nasal patella by cutting vertically from the first molars of the maxilla with the saw.In this way, measure the gap between the nasal patella and the facial bone with a ruler to indicate the degree of nasal patella atrophy. . If the gap between the nasal patella and the facial bone is less than 5mm, it is considered normal, and if it is 6-8mm, light atrophy, 9-11mm is moderate, and more than 12mm is severe. If the secondary infection is severe, the symptoms of pneumonia are severe.

Difficulty in controlling atrophic rhinitis (AR) has high expression rate. Bodetella bronchiceptica bacteria are susceptible to drug resistance because of the temporary transition from non-toxic to strong-tolerant or vice versa depending on environmental conditions.

Therefore, intranasal nebulization is administered to mammal pigs. In Korea, antibiotics should be selected after drug resistance test. In fact, in Korea, kanamycin and gentamicin are generally used because they are good.

An object of the present invention is a composition for spraying pig nasal to remove Bodetella bronchiseptica bacteria and secondary infection bacteria Pasteurella multocida D bacteria in the nose for absolute eradication of swine atrophic rhinitis To provide.

The present invention for the prevention and treatment of swine atrophic rhinitis

Amikacin and its pharmaceutically acceptable salts, and beta-glucan as an active ingredient characterized in that it provides a composition for spraying pig nasal.

Pig nasal spray composition according to the present invention can be eradicated by atrophy of rhinitis rhinitis (AR) as the drug progresses for a long time by spraying in the nasal cavity, including amikacin and beta-glucan.

Pig nasal spray composition according to the invention is used for the prevention and treatment of swine atrophic rhinitis.

The composition comprises amikacin and its pharmaceutically acceptable salts and beta-glucan as an active ingredient.

Amica of compounds represented by the following general formula (1) God, (2 S) -4- amino -N- [(2 S, 3 S , 4R, 5 S) -5- amino -2 - [(2 S, 3 R , 4 S, 5 S , 6 R ) -4-amino-3,5-dihydroxy-6- (hydroxymethyl) oxan-2-yl] oxy-4-[(2 R , 3 R , 4 S , 5 R , 6 R ) -6- (aminomethyl) -3,4,5-trihydroxy-oxan-2-yl] oxy-3-hydroxy-cyclohexyl] -2-hydroxy-butaneamide { (2 S ) -4-amino-N-[(2 S , 3 S , 4 R , 5 S ) -5-amino-2-[(2 S , 3 R , 4 S , 5 S , 6 R )- 4-amino-3,5-dihydroxy-6- (hydroxymethyl) oxan-2-yl] oxy-4-[(2 R , 3 R , 4 S , 5 R , 6 R ) -6- (aminomethyl) -3 , 4,5-trihydroxy-oxan-2-yl] oxy-3-hydroxy-cyclohexyl] -2-hydroxy-butanamide}:

[Formula 1]

Conventionally, kanamycin and gentamicin, which are known to have good efficacy, are most widely used for swine atrophic rhinitis. However, the present inventors, according to their experiments pigs causing atrophic rhinitis Bode telra chevron kisep urticae (Bordetella bronchiseptica) bacteria and a secondary pathogen in wave stew Pasteurella far Toshio dagyun effectively atrophic rhinitis it is resistant much about (Pasteurella multocida) D hyeonggyun blossomed It is not healing.

Amikacin is 100% resistant to Bodetella bronchiseptica and secondary infection bacterium Pasteurella multosidida, and can be effectively used for the prevention and treatment of atrophic rhinitis.

Such amikacin may be used in the form of an animal-pharmaceutically acceptable salt for application in combination with the compound.

The animal pharmaceutically acceptable salts include alkali metal salts such as lithium, sodium, potassium salts; Alkaline earth metal salts such as calcium or magnesium salts; And salts consisting of suitable organic ligands such as tetravalent ammonium salts. In the case of acid addition salts, for example, a compound solution according to the invention can be formed by mixing with a pharmaceutically acceptable non-toxic acid solution such as hydrochloric acid, fumaric acid, maleic acid, succinic acid, acetic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid. Can be. Such acids are used to the extent that they do not significantly irritate the nasal tissue that the composition contacts.

In addition, the composition for spraying nasal pigs according to the present invention includes beta-glucan, which is an immune enhancer, to increase the efficacy of the amikacin.

Beta-glucan (β-glucan) activates the immune function of normal cellular tissues, stimulates the release of a large number of immune-enhancing substances in the pig's body, and has the effect of strengthening the pig's immunity by strengthening the resistance to viruses . In addition, the beta-glucan has the effect of preventing antibiotic abuse due to the above-mentioned effect.

Pig nasal spray composition according to the present invention is used by diluting amikacin with water in a volume ratio of 10 to 1000 times. Preferably, 5-20 ml, preferably 10 ml, of beta-glucan is used for 10 ml of 10-fold dilutions of amikacin.

If the content of amikacin and beta-glucan is out of the above range, the sterilization effect on Bodetella bronchiceptica bacteria, a cause of atrophic rhinitis, and Pasteurella multocida bacteria, secondary infection bacteria, is reduced. Use suitably within the range.

The composition for spraying a pig nasal cavity according to the present invention comprising the above composition is prepared in the form of a liquid formulation for administration to the nasal cavity. Such compositions are prepared and used at a suitable concentration to spray or dissolve an appropriate amount of amikacin and beta-glucan in water.

At this time, the composition for spraying pig nasal includes an electrodeposition agent to increase the persistence of the drug in the nasal cavity.

The electrodeposition agent serves to increase the permeability of amikacin and beta-glucan into the nasal cavity and to enhance the intranasal adhesion. As the electrodeposition agent, a known material may be used, and is not particularly limited in the present invention. For example, soap and sodium bicarbonate, a carba liquid (mixture of sodium salts of alkylacyl polyethoxylates, alkyl sulfonate alkylates), medicaments (from GROW MORE, USA) and mixtures thereof can be used.

The electrodeposition agent is diluted with water in a volume ratio of 100 to 5000 times with water, preferably 0.2 to 10 ml, preferably 0.25 ml is added per 1 L of water.

In this case, when the electrodeposition agent is used in the nasal spray composition, 5-20 ml of beta-glucan, preferably 10 ml, is used for 10 ml of dimycincin diluted 10-fold, and the electrodeposition agent is 0.25 ml (diluted with 1 L of water). use.

In addition, the composition for pig nasal spray according to the present invention may be used additives commonly used in the art. Such additives depend on the desired form and the delivery device used. In one example, the additive may include a pharmaceutically acceptable carrier comprising a cosolvent such as ethanol, propylene glycol, glycerol, a water miscible solvent; Liquid aerosol propellants and mixtures thereof.

A preservative may also be further included to prevent microbial contamination of the dosage device or composition applied to the nose. The preservatives include topical nasal compositions including benzalkonium chloride, chlorhexidine gluconate, phenyl ethyl alcohol, phenoxyethanol, benzyl alcohol, sorbic acid, benzoic acid, thimerosal, phenylmercuric acetate, and combinations thereof. It usually includes related things.

Pig nasal spray composition according to the present invention can be administered as a nebulizer, inhaler, drop, and can also be administered to the nasal cavity of the pig in a nasal spraying manner by a well-known suitable delivery system.

Nasal sprays are fast absorbing and have high bioavailability, allowing small doses to be administered. It is also fast to express action after spraying, suppresses metabolism in the liver or gastrointestinal tract, and avoids gastrointestinal mucosal irritation. And because it is used in a diluted state, there is a low risk of being administered in an overdose and it is convenient to use.

Such a composition for spraying nasal pigs is particularly effective in killing Bodetella bronchiseptica and the second infectious bacterium Pasteurella multocida in the pig's nose.

Hereinafter, the present invention will be described in more detail with reference to the following examples, which are illustrative only for explaining the present invention, and the present invention is not limited thereto.

Experimental Example 1: Resistance Test

Tolerance test against Bordetella bronchiseptica , which causes swine atrophic rhinitis, and Pasteurella multocida , a secondary infection, against drugs used as various antibiotics and amikacin And the results are shown in Table 2 below.

Pharmaceutical / Isolates Bordetella bronchiseptica Pasteruella moltocida Good pharmaceutical / isolate ratio Good pharmaceutical / isolate ratio Amikacin (AN) 89/89 100.00% 50/50 100.00% Penicillin (P) 0/89 0.00% 1/50 1.96% Ampicillin (AM) 2/89 2.25% 3/50 5.88% Amoksacillin (AmC) 65/89 73.03% 44/50 86.27% Colistin (CL) 78/89 87.64% 46/50 90.20% Kanamycin (K) 12/89 13.48% 19/50 37.25% Gentamicin (GM) 22/89 24.72% 9/50 17.65% Neomycin (NEO) 15/89 16.85% 26/50 50.98% Streptomycin (S) 0/89 0.00% 1/50 1.96% Sulfamethoxazole + trimethoprim
(SXT) 18/89 20.22% 16/50 31.37% Ciprosin (CIP) 35/89 39.33% 33/50 64.71% Noflo sacin (NOR) 37/89 41.57% 33/50 64.71% Seti Warper (XNL) 47/89 52.81% 24/50 47.06% Sepharosporin (CF) 18/89 20.22% 31 60.78% Tetracycline (Te) 0/89 0.00% 0/50 0.00% Doxycycline (D) 1/89 1.12% 0/50 0.00% Lincosfectin (LIN + SP) 31/89 34.83% 11/50 21.57%

Referring to Table 2, 100% of amikacin compared to kanamycin (13.48% and 37.25%) and gentamicin (17.65% and 24.72%), which are known to have good efficacy in conventional swine atrophic rhinitis. It can be seen that it is effective.

Experimental Example 2

In the resistance test, 100 mg of amikacin was added to 1 ml of water, and even when diluted to 1,000-fold, 100% bactericidal effect was observed in both Bodetella bronchiceptica and the second infectious bacterium D. bacterella multocida. It can be seen that even if diluting at a high magnification, the bactericidal effect of amikacin is maintained.

Experimental Example 3

Five nasal spray compositions were prepared. On the Muller-Hinton Agar plate to which the respective compositions were added, Bordetella bronchiseptica bacteria 2 × 10 ⁸ , CEM cells / ml, and the second infectious bacterium Pasteurella multocida ( pasteurella) multocida ) After inoculating CEM cells / ml into D-type 4D 2 × 10 ⁸ , respectively, it was visually observed whether colonies were formed after incubation at 37 ° C. The inoculation was carried out three times at monthly intervals.

Composition A: 10 ml 10-fold diluted amikacin

Composition B: 10 ml diluted 10-fold amikacin + 10 ml beta-glucan

Composition C: 10-fold dilution amicacin 10ml + drug-taking 10ml (medicine-taking 0.25 ml + distilled water 1L)

Composition D: 10-fold diluted Amikacin 10ml + Beta-Glucan 10ml + Drug Extraction 10ml (Approximate Drug 0.25ml + Distilled Water 1L)

Composition E: 10 ml of distilled water

Figure 2 (a) is Bordetella bronchiseptica bacteria according to the composition for nasal spray, (b) after the first inoculation of D-type bacterium Pasteurella multocida (secondary infection) It is a photograph. 3 is a photograph after the second inoculation, and FIG. 4 is a photograph after the third inoculation.

Referring to Figures 2 to 4, it can be seen that the growth of bacteria in the compositions A to D containing amikacin is inhibited, the growth of the bacteria was observed only in the composition E using distilled water.

Experimental Example 4

The following four nasal spray compositions were prepared, followed by intranasal spraying of 5 pigs for 20 pigs.

Composition 1: commercial kanamycin intranasal spray (sulfuric acid kanamycin 40 mg / 1 ml)

Composition 2: Commercial Gentamicin Intranasal Spray Solution (Gentamicin Sulfate 100mg / 1ml)

Composition 3: 10-fold dilution amicacin 10ml + beta-glucan 10ml

Composition 4: 10-fold diluted Amikacin 10ml + Beta-Glucan 10ml + Drug Extraction 10ml (Approximate Drug 0.25ml + Distilled Water 1L)

Kanamycin or gentamicin were administered intranasally at 1 day (within 8 hours), 1 week, 2 weeks, 3 weeks, and 4 weeks of age. In addition, in the case of a composition using amikacin, a method was employed to make one day (within 8 hours), 3 days, 10 days, and weaning.

5 is a schematic diagram showing the nasal cross-sectional state of the pig according to the degree of atrophic rhinitis. As shown in FIG. 5, when the two patellas atrophy, it is regarded as having a severe rhinitis.

Figure 6 is a photograph showing the nasal cross-section of the pig sprayed with a composition 1 containing kanamycin, Figure 7 is a photograph showing the nasal cross-section of the pig sprayed with a composition 2 containing gentamycin, Figure 8 is 10 A photograph showing the nasal cross-section of a pig sprayed with Composition 3 containing 10 ml of pear diluted amikacin 10 ml + 10 ml of beta-glucan (omitted-amicincin + beta-glucan), FIG. 9 is a 10 ml of 10 ml diluted 10-fold amikacin The nasal cross-sectional state of pigs sprayed with Composition 4 (omitted-1) comprising +10 ml of beta-glucan + 10 ml of drug-taking (0.25 ml of drug-taking + 1 L of distilled water) (omitted-amicinin + beta-glucan + electrodeposition agent) It is a picture showing.

Compositions containing kanamycin (composition 1) or gentamicin (composition 2) need to catch pigs and spray them intranasally. Iron injections must be taken at 3 and 10 days of age, so they need to catch pigs. have. 6 and 7, the complete disappearance of the nasal cavity was shown when kanamycin or gentamicin were used.

In comparison, as shown in Figs. 8 and 9, the compositions 3 and 4 using amikacin showed good formation of the nasal patella, which concluded that the composition could be used immediately.

Figure 1 is a schematic diagram showing the nasal cross-section of healthy pigs and pigs with atrophic rhinitis.

Figure 2 (a) is Bordetella bronchiseptica bacteria according to the composition for nasal spray, (b) after the first inoculation of D-type bacterium Pasteurella multocida (secondary infection) It is a photograph.

The (a) is a Bode telra chevron kisep urticae (Bordetella bronchiseptica) according to the nasal spray composition of the 3 fungi, (b) a secondary pathogen in wave stew Pasteurella far Toshio dagyun (Pasteurella multocida) after the second inoculation of D hyeonggyun It is a photograph.

Figure 4 (a) is Bordetella bronchiseptica (b) bacteria according to the composition for nasal spray, (b) after the third inoculation of D-type bacteria Pasteurella multocida (secondary infection) It is a photograph.

5 is a schematic diagram showing the nasal cross-sectional state of the pig according to the degree of atrophic rhinitis.

Figure 6 is a photograph showing the nasal cross-sectional state of the pig sprayed with composition 1 containing kanamycin.

Figure 7 is a photograph showing the nasal cross-sectional state of pigs sprayed with composition 2 containing gentamicin.

Figure 8 is a photograph showing the nasal cross-sectional state of pigs sprayed with Composition 3 comprising amikacin + beta-glucan.

Figure 9 is a photograph showing the nasal cross-sectional state of pigs sprayed with composition 4 comprising amikacin + beta-glucan + electrodeposition agent.

Claims (6)

For the prevention and treatment of swine atrophic rhinitis, Amikacin and a pharmaceutically acceptable salt thereof, and a composition for spraying pig nasal, characterized in that it comprises beta-glucan as an active ingredient. The method of claim 1, The nasal spray composition is a pig nasal spray composition, characterized in that it has a bactericidal effect against Bodetella bronchiseptica ( Bacterella bronchiseptica ) and secondary infection bacteria Pasteurella multocida (D) bacteria. The method of claim 1, The amikacin composition for spraying pig nasal, characterized in that diluted to 10 to 1000 times the water (volume ratio) used. The method of claim 1, The nasal spray composition is a pig nasal spray composition, characterized in that containing 0.5 to 20 ml of beta-glucan per 1 ml of dimycin diluted 10-fold. The method of claim 1, In addition, the composition for nasal spray further comprises a composition for spraying pig nasal, characterized in that it further comprises an electrodeposition agent. The method of claim 5, The nasal spray composition for pig nasal spray comprising 5 to 20 ml of beta-glucan, and 0.2 to 10 ml of electrodeposition agent (diluted with 1 L of water) with respect to 10 ml of dimycincin diluted 10-fold.

Images