WO2004024151A1

WO2004024151A1 - Composition and potentiating method

Info

Publication number: WO2004024151A1
Application number: PCT/SE2002/001640
Authority: WO
Inventors: Shoaá ABDUL-RAHMAN
Original assignee: New Pharma Research Sweden Ab
Priority date: 2002-09-13
Filing date: 2002-09-13
Publication date: 2004-03-25
Also published as: AU2002337534A1; BR0213235A; MXPA05002619A; CN1638770A

Abstract

A composition is described, which comprises clopidol (3,5-dichloro-2, 6-dimethyl-4-pyridinol) or its analogue dinitolmide (2-methyl-3,5-dinitrobenzamide) in a mixture with a potentiating agent comprising at least one of colistin, benzoic acid or derivatives thereof, salicylic acid or derivatives thereof, sulfaquinoxaline (4-amino-N-2-quinoxalinylbenzenesulfonamide) or amantadine (tricyclo-[3.3.1.1]decan-1-amine). The composition may be used as a medicament, a potentiating method or for controlling fungi in animals. The composition has anti-coccidiocidal effect as well as prophylactic effect on coccidiosis.

Description

COMPOSITION AND POTENTIATING METHOD

The present invention relates to a novel composition, a composition for use as a medicament, especially for treating coccidiosis, and use of a composition for the manufacture of a drug for treating deseases cused by fungi in animals.

Coccidiosis is a disease of the intestinal lining of poultry, for example chickens, caused by protozoan cocci- dial parasites of the genus Eimeria, such as Eimeria te- nella, Eimeria necatrix, Eimeria acervulina, Eimeria bru- netti, Eimeria maxima, Eimeria mitis, Eimeria mivati, Eimeria praecox and Eimeria hagani . Species belonging to said genus cause clinical disease in chickens and immunity to any one species does not protect birds against infection with other species. Coccidiosis is seen most com- monly in chickens between 3 and 6 weeks of age. Coccidiosis is caused by protozoa of the family Eimeriidae. Various sites in the intestine are infected. The infectious process is rapid (4-7 days) and is characterised by parasite replication in host cells with extensive damage to the intestinal mucosa. Clinical disease occurs only after ingestion of relatively large numbers of sporulated oocysts by susceptible birds. Both clinically infected and recovered birds shed oocysts in their droppings, which contaminate feed, dust, water, litter and soil. Anticoccidial drugs are available on the market for prevention and treatment of coccidiosis in especially chickens and turkeys. Anticoccidials are generally given to poultry in the feed to prevent acute disease. Water medication is generally preferred over feed medication for treatment. Antibiotics are sometimes used to improve rate of recovery and prevent secondary infections.

Clopidol, i.e. 3 , 5-dichloro-2 , 6-dimethyl-4-pyridi- nol, and its analogue Dinitolmide, i.e. 2-methyl-3 , 5-di- nitrobenzamide are today the most frequently used anti- coccidial drugs. There is a need in the art to provide a drug having enhanced effect in the treatment of coccidiosis. This need is met according to the invention by provision of a composition comprising besides Clopidol and its analogue at least one potentiating component selected from the group consisting of Colistin, benzoic acid or derivatives thereof, salicylic acid or derivatives thereof, 4-amino-N-2-quinoxalinylbenzenesulfonamide and tri- cyclo (3.3.1.1. ) decan-1-amine . Said components are named potentiating agents. The mechanism, by which the enhanced activity is obtained, is not for the present understood. Tests have shown, that enhanced activity is obtained, and reference is made to test results presented in this specification. Further, according to the invention, it has quiet unexpectedly been found, that an antifungal effect is achieved when the composition is used as a drug for treating for example poultry.

SUMMARY OF THE INVENTION

In a first aspect the invention relates to a compo- sition as claimed in claim 1.

In a second aspect the invention relates to a composition for use as a medicament as claimed in claim 2.

In a third aspect the invention relates to a potentiating method as claimed in claim 3. In a fourth aspect the invention relates to use of a composition of the kind defined above for the manufacture of a drug for treating deseases caused by fungi in animals. According to this aspect, the animals comprise for example pultry. Below is a brief description of Clopidol and its analogue and the potentiating agents contained in the composition. Benzoic Acid Benzoic acid has the formula

Benzoic acid is used for preservation and for manufacture of soap, perfume and pigments. It is present in almost all berries in amounts of 0,05%, where it functions as a natural preservative. Yoghurt may contain up to 30 mg/kg.

It is, according to the invention, possible to use the following derivatives of benzoic acid instead of benzoic acid. Benzoic acid derivatives

3-acetamido-5-aminobenzoic acid

2-acetamido-2-aminobenzoic acid

4-acetamidobenzoic acid

4-acetamidobenzoic acid methyl ester 4-acetamido-N-butyl benzoic acid

2-acetamido-5-chlorobenzoic acid methyl ester 4-acetamido-5-chloro-2hydroxy benzoic acid methyl ester 4-acetamido-2-hydroxy benzoic acid 4-acetamido-2-methoxy benzoic acid 2-acetamido-4-nitrobenzoic acid 4-acetoxybenzoic acid 2-acetylbenzoic acid 5-acetyl-2-hydroxy benzoic acid 4-allylbenzoic acid ethyl ester 3-aminobenzoic acid

3-amino-5- (aminosulfonyl) -4-phenoxy benzoic acid 4-aminobenzoic acid butyl ester 4-aminobenzoic acid ethyl ester 4-amino-2-butoxy benzoic acid 2-amino-3-chloro benzoic acid

4-amino-3 , 5-dichloro benzoic acid 2-amino-4, 5-difluoro benzoic acid 2 -amino-4 , 5-dimethoxy benzoic acid

2-amino-5-fluoro benzoic acid

2-amino-3-hydroxy benzoic acid

3-amino-4-hydroxy benzoic acid 2-amino-6-methoxy benzoic acid

4-amino-3-methyl benzoic acid

4- (aminomethyl) -benzoic acid

4-amino-3-nitro benzoic acid

2-amino-4-sulfo benzoic acid 5-aminosulfonyl-2 , 3 -dimethoxy benzoic acid

4-amylbenzoic acid

2-anilinobenzoic acid

Benzoic acid 2 methylbutyl ester

4-benzoyl benzoic acid 5-benzyl-2-mercaptobenzoic acid

2- (benzyloxy) -benzoic acid

2 , 2 ' -dibenzoic acid

4 , 4 ' -dibenzoic acid

4-bromobenzoic acid 3-bromo-2 -6-dimethoxy benzoic acid

4-butoxybenzoic acid

4-tert-butylbenzoic acid

4 -carbmethoxy-3 , 5-dimethoxy benzoic acid

4-chloro-2-nitrobenzoic acid 4-cyanobenzoic acid-4-propylphenyl ester

3- (1-cyanoethyl) -benzoic acid

3 , 5-diaminobenzoic acid

4- (diamylamino) -benzoic acid

3 , 5, di-tert-butylbenzoic acid 2 , 6-difluorobenzoic acid

2 , 5-dihydroxy benzoic acid

2 , 6-dimethoxy benzoic acid

3 , 5-dimethoxy-4-hydroxybenzoic acid

2 , 4-dimethylbenzoic acid 3 , 5-dimethyl-4-hydroxybenzoic acid

4-ethoxybenzoic acid

4-formylbenzoic acid 4-guanidinobenzoic acid

4-heptylbenzoic acid

4-hydrazinobenzoic acid

4-hydroxybenzoic acid 4-isothiocyanato benzoic acid

4-mercapto benzoic acid

4-phenyl benzoic acid

4- (phenylthio) -benzoic acid

4-propoxy benzoic acid selenino benzoic acid

4-sulfo benzoic acid

4-sulfonamido benzoic acid

3 , 4-5-6-tetrahydro benzoic acid thiobenzoic acid trimethoxy benzoic acid Salicylic acid

This has the formula

and is a weak acid used for relief of pain, disinfection and as febrifuge. It inhibits the enzyme cyclooxygenase that stimulates the production of prostaglandines, which have pain-, fever- and inflammationpromoting effects. In some skin diseases salicylic acid is applied to the skin as a keratin-dissolving drug. Clopidol

Clopidol (3 , 5-dichloro-2, 6-dimethyl-4-pyridinol) is known as an agent in the prevention of coccidiosis in chickens and turkeys. It is administered orally by addition to the feed.

Dinitolmide

Dinitolmide or 2-methyl-3 , 5-dinitrobenzamide has the formula

and is used in veterinary medicine as a coccidiostat . Amantadine Amantadine (tricyclo [3.3.1.1] decan-1-amine) is used as an antiparkinsonian although the mode of action is not completely known. It effects the incapability of moving, the stiffness and the shakings. Amantadine is also known to have some antiviral effect on influenza virus type A, as it inhibits the propagation of virus in the attacked cells.

Colistin

Colistin is a cyclopolypeptide antibiotic produced by Bacillus colistinus isolated from Japanese soil. It is used as an antibacterial agent for both humans and animals .

Sulfaquinoxaline (QS)

Sulfaquinoxaline or 4-amino-N-2-quinoxalinylbenzene- sulfonamide has the formula

and is used as a coccidiostat. Other sulfonamides can be used According to a preferred embodiment Roxarsone is used in the composition (not as a potentiating agent) .

Roxarsone or 4-hydroxy-3-nitrophenylarsonic acid has the following formula

It is used as an antibacterial and in veterinary medicine to control enteric infections and improve growth and feed efficiency.

Examples of concentrations and carriers . It is the treating veterinary' s choice to decide the concentrations suitable for treatment. However, it is re- commended that the concentration of Clopidol and that of its analogue according to the invention should be at least 20 ppm and at most 125 ppm. The concentration of benzoic acid or its derivatives or salicylic acid or derivatives thereof should range from 9-110 ppm, the con- centration of Sulfaquinoxaline should range from 9-55 ppm, the concentration of Roxarsone should range from 5- 15 ppm, the concentration of Amantadine should range from 4-27 ppm and the concentration of Colistin should range from 4-100 ppm. The concentrations of the compositions are based on the feed or drinking water preparations ad lib, i.e. for free feed or drinking water consumption during a normal practical fattening or rearing period. All feed formulations conventional in the poultry industry are suitable as carriers for the compositions according to the invention. Examples are feedstuffs based on some types of cereal grains which contain vitamin concentrates, mineral concentrates or other active substances and feed additives in any concentration. Both conventional dry mealy or pelletised feedstuffs but also liquid feed suspensions including feedstuffs such as distillers' residues and milk by-products can be used in the case of the compositions according to the invention.

To prepare the poultry feed a concentrated premix is usually first made which contains Clopidol or its analo- gue and at least one of the components of the composition in high concentration, for example 0.2-75% by weight. For this purpose the components are either dispersed or mixed in inert carrier substances, such as vermiculite, diato- maceous earth, attapulgite, calcium carbonate, etc., to- gether with physiologically harmless carriers, such as propylene glycols, polyethylene glycols, inert oils, such as vegetable oils, highly refined mineral oils, ethanol , water, or aqueous alcohols. Organic carrier material, such as wheat bran, shredded maize, soybean flour, lucer- ne meal, rice husks or ground maize cobs and any other organic carrier substances from vegetable products are likewise suitable for this purpose.

As mentioned above, the compositions according to the invention may further also be administered to poultry together with the drinking water. Their inclusion in the drinking water is achieved by adding a form of the composition concerned, which is soluble in water or can be suspended in water to the drinking water in a suitable quantity. Such preparations are in general produced by selecting a water-soluble form of the composition. Water- insoluble forms, for example suspensions, may also be used. In such cases physiologically harmless auxiliary agents which keep the composition according to the invention in suspension in water over a prolonged period are used. Auxiliary agents, which are suitable for this purpose, are swelling agents, such as alginates, gelatine, carboxymethylcellulose or polyvinylpyrrolidone . Surfactant compounds, such as for example naphthalenesulfona- tes, alkylbenzenesulfonates, or polyoxyethylene sorbitan esters may be used for bringing the compositions into suspensions. Normally, a concentrated suspension or a dry formulation of the composition according to the invention and the suspension agents is produced in certain mixing ratios and is then diluted with the drinking water to the required application concentration or the premixes are mixed in suitable concentrations with the feedstuffs nor- mal in practice. The drinking water or feed so medicated is then fed to the poultry either initially ad lib or for a certain period.

Although the invention has been described above with particular reference to the treatment of poultry against coccidia infections, treatment of other domestic and useful animals, for example other birds, rabbits, pigs and ruminants is also within the scope of the invention. Biological Examples Anticoccidiocidal activity Materials and Methods Chickens

One day old Habbared X breed chickens were used in the tests. In the 7 different tests the chickens were allocated into 10 equal groups (20-30 birds in each group) , one group serving as a control group and one as a positive control group. The control group was infected but received no treatment, the positive control group was also infected and received a known, reference anti-coccidial drug (Clopidol 125 ppm or Salinomycin 60 ppm) . The chickens were reared in 1*1 m pens (each in its group) on medicated feed from the first day of their lives to day 15. Each group in each test was given a different mixture of Clopidol together with at least one component selected from the group consisting of Colistin, benzoic acid or derivatives thereof, salicylic acid or derivatives thereof, Sulfaquinoxaline, and Amantadine. On the 16^th day the chickens were infected orally with a mixture of 6000-8000 mature sporulated field collected oocysts, containing different Eimeria strains; Eimeria acervulina, Eimeria maxima, Eimeria necatrix, Eimeria tenella and Eimeria brunetti. Fresh faecal droppings were collected daily from the 5^th day post experimental infection until the 10^th day post infection. The oocysts were counted and the mean number of oocysts per gram faeces for each group was cal- culated using the Mc-Master technique according to Souls- by (1984) . The number of dead birds per day was also counted and the cause of death was recorded after a postmortem examination.

Medication groups : Each composition to be tested was mixed in a sufficient amount of feed for each group and was used for daily feeding from the day zero of the test. Fresh water was supplied ad-libetum.

Evaluation of the efficacy The efficacy of the prophylactic effect was evaluated by comparing the total number of shed oocysts in the medicated groups with the number in the non-medicated group during the whole period of the experiment . The weight of the birds as well as the amount of consumed feed were determined at the end of the experiment. From this the mean body weight and mean amount of consumed feed per bird were calculated. The results are shown in the tables below.

As for the treatment-column, the composition of the medicament is given in ppm. Appropriate abbreviations are used.

Table (1) : Prophylactic effect of products on experimental coccidiosis in broiler chicken on feed . (Ref . May 2001)

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C CD </>

m m m c 7i m

σ> d.p.i. = days post experimental infection

Table (1) : Prophylactic effect of products on experimental coccidiosis in broiler chicken on feed. (Ref. April 2001)

c

CD

m m m

7i c m σ>

d.p.i. = days post experimental infection.

Table (1) : Prophylactic effect of products on experimental coccidiosis in broiler chicken on feed . (Ref . Feb 2001)

</>

C CD </>

m m m c 7i m σ>

d.p l. = days post experimental infection.

Table (1) : Prophylactic effect of products on experimental coccidiosis in broiler chicken on feed. (Ref. December 2000)

c

CD

m m m

7i c m σ>

d.p.i = days post experimental infection.

Table (1) : Prophylactic effect of products on experimental coccidiosis in broiler chicken on feed ( Ref. October 2000).

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C CD </>

m d.p.i. = days post experimental infection

7i

C m σ>

Table (1) : Prophylactic effect of products on experimental coccidiosis in broiler chicken on feed ef. July 2000.

I\J σ>

c

CD

m m m c 7i

d.p.i. = Days post expeπmental infection. , *** mortalities m these birds were related to infection and other causes than coccidia m ***** The group reared in cages and the same treatment as that of group 2 σ>

Table (1) Testing the anti-coccidiocidal effect of new compounds in young birds (dose 25-30 000 Mixed oocysts of Egyptian strain) (Ref. May 2000)

c

CD

m

d.p.i. = Days post experimental infection. *** mortalities m these birds were related to infection and other causes than coccidia m m

7i

C

I- m

I\J σ>

Antifungal effect

The following compositions were tested. Toxy-nil is produced by Nutri-AD Company, Belgium Mycobond is a mycotoxin binding agent . Both are used herein as controls.

[1] I gram of 50 clopidol + 36coIistin + 14 benz (1004). ,..(1) not testedas anticoccidial.

[2] I gram of 80 clopidol + 5 5 colistin + 15 b aiz (504).(2).not tested for coccidia.

(3) 1 gram of 120 clopidoϊ + 90 colistin ÷ 40 bem. (904) (3) has good anticoccidial

14] I gram of 50 clopidol + 40 colistin + 40 benz (204) (4) has good anticoccidial

[5] I gram of 60 clopidol + 45colistin + 35 benz (304) (5) not tested for coccidia.

[6] I gram of 100 clopidol + 71 colistin + 29 benz (704) (ό)not tested for coccidia, dilution

Claims

1. A composition comprising 3 , 5-dichloro-2 , 6- dimethyl-4-pyridinol or its analogue 2 -methyl-3 , 5- dinitrobenzamide in a mixture with at least one component selected from the group consisting of Colistin, benzoic acid or derivatives thereof, salicylic acid or derivatives thereof, 4-amino-N-2-quinoxalinylbenzenesulfonamide and tricyclo [3.3.1.1] decan-1-amine .

2. A composition for use as a medicament comprising 3 , 5-dichloro-2 , 6-dimethyl -4 -pyridinol or its analogue 2- methyl-3 , 5-dinitrobenzamide in a mixture with at least one component selected from the group consisting of Colistin, benzoic acid or derivatives thereof, salicylic acid or derivatives thereof, 4-amino-N-2-quinoxalinyl- benzenesulfonamide and tricyclo [3.3.1.1] decan-1-amine, 3 , 5-dichloro-2 , 6-dimethyl-4 -pyridinol or its analogue 2- methyl-3 , 5-dinitrobenzamide and said components being present in therapeutically effective amounts and optionally in admixture with therapeutically acceptable carriers .

3. Method for potentiating 3 , 5-dichloro-2 , 6- dimethyl-4-pyridinol or its analogue 2 -methyl -3 , 5- dinitrobenzamide comprising treating 3 , 5-dichloro-2 , 6- dimethyl-4 -pyridinol or its analogue with at least one component selected from the group consisting of Colistin, benzoic acid or derivatives thereof, salicylic acid or derivatives thereof, 4-amino-N-2-quinoxalinylbenzenesul- fonamide and tricyclo [3.3.1.1] decan-1-amine .

4. Use of a composition according to claim 1 for the manufacture of a drug for treating deseases caused by fungi in animals.