FR2605221A1 - Pharmaceutical or veterinary compositions containing 25-de(hydroxymethyl)-25-deoxy-25-oxomonensin - Google Patents

Abstract

The invention relates to pharmaceutical or veterinary compositions containing, as essential active principle, 25-de(hydroxymethyl)-25-deoxy-25-oxomonensin or one of its non-toxic salts, in combination with a suitable vehicle, diluent or excipient.

Description

PHARMACEUTICAL OR VETERINARY COMPOSITIONS CONTAINING
DE (HYDROXYMETHYL) -25 DEOXY-25 OXO-25 MONENSIN.

 The present invention relates, in general, to pharmaceutical, veterinary or food compositions useful in particular for combating coccidiosis.

laquelle sera désignée par la suite sous l'appellation "monensine lactone", ainsi que ses sels non toxiques.In particular, the invention relates to pharmaceutical, veterinary or food compositions containing, as essential active principle, de (hydroxymethyl) -25 deoxy-25-oxo-25-monensin of formula

which will subsequently be referred to as "monensin lactone" and its non-toxic salts.

Non-toxic salts are more particularly alkali metal salts, such as lithium, sodium or potassium, alkaline earth metal salts, such as calcium or magnesium or ammonium salts.
such as those obtained from ammonia or an amine such as methylamine, ethylamine, dimethylamine, diethylamine, triethylamine, ethanolamine, diethanolamine and the like.

Coccidiosis is a common and widespread disease in poultry, pigs, rabbits, cats and dogs, in some ruminants, etc., a disease caused by parasites belonging to the branch of protozoites of the genus Eimeria, such as E.tenella, E.necatrix, E.acervulina,
E.maxima, E.brunetti, E.mivati, E.adenoldes and E.maleagrimitis.

 E.tenella and E.acervulina are the causative agents of the most severe and often fatal forms of this disease. They are characterized by severe and severe haemorrhage, accumulation of blood in the cecum and passage of blood through the feces.

 E.necatrix attack. the small intestine of the chicken causing intestinal coccidiosis while E. maleagrimitis and E. adenoldes are responsible for coccidiosis in turkeys.

 Left untreated, coccidiosis leads to less than normal weight gain, reduced feed efficiency and high mortality in birds.

 As a result, morbidity and mortality from coccidian infestations create a significant economic loss when left untreated or unattended.

 Elimination or control of this disease is therefore of prime importance, especially for poultry farming.

 In the context of the present invention, it has been found that monensin lactone and its non-toxic salts have very interesting anticoccidial properties, in particular with respect to Eimeria tenella, which can be used for the destruction of parasites in birds. and some ruminants (sheep).

 In addition, it has surprisingly been found that monensin lactone and its non-toxic salts, while exhibiting anticoccidial activity equivalent to that of monensin, are much less toxic than this compound.

 As an anticoccidial agent, monensin lactone will therefore have an undeniable advantage over monensin.

 In addition, it will be mentioned that monensin lactone is capable, in combination with certain antibiotics, in particular antibiotics aminoglycosides, to potentiate the action of these compounds.

 As such, monensin lactone and its non-toxic salts, in combination with certain antibiotics, could be used in human medicine.

 Pharmacological tests have been carried out in order to highlight the properties which, taken as a whole, are capable of rendering monensin lactone and its non-toxic salts useful in particular as anticoccidial agents.

I. Anticoccidial activity
Tests were carried out in which the zootechnical results (weight, consumption index) and the clinical symptoms of coccidiosis (morbidity, mortality, faecal appearance, ectocystal excretion, lesions) were recorded.

The following experimental protocol was used for this purpose
15-day-old Warren Sex Sal Link genetic strain cockerel flocks are divided into 4 groups of 4 animals each and fed with a broiler type feed. In addition, a compound to be studied is administered to animals of 2 groups, namely 100 ppm / day of monensin lactone or 100 ppm / day of monensin.

 After 48h (day OJ) 250 of the sporulated oocysts of Eimeria tenella were inoculated with the animals of the 2 groups which received a compound to be studied as well as those of a third group.

 The feeding and the treatment initiated for each of the 4 groups are then continued for 8 days after inoculation of the oocysts.

Animals are weighed, sacrificed and examined for coccidia lesions.

The following table reports all the results obtained with the following groups
NCNT: uncontaminated, untreated animals
CTNT: contaminated animals, untreated
CTMO: contaminated animals treated with monensin
CTML: Contaminated animals treated with monensin lactone on the following criteria determined during the overall test period
- Weight gains (GP) in g determined from D-3 to D + 8 (P '= weight)
in g to J-3, P = weight in g to D + 8)
- Daily ookysteal excretion (EO) from D + 5 to D + 7 - Morbidity (M) noted from 0 to 4, namely
O: normal attitude of animals
1: ruffled feathers (especially those of the neck)
2: beginning of reluctance and prostration
3: reluctance and prostration are more marked
and the animals are sad
4: the animals whose feathers are ruffled have
a "ball" position, the wings are falling,
the standing is painful to them and they have the
eyes closed.

- The aspect of faeces (MF) noted from to 4 namely
O: normal
1: traces of bleeding
2: some blood stains
3: many blood spots
4: very haemorrhagic - Mortality (M2) expressed as a percentage - lesions (L) noted according to REID notation (W.Malcolm REID,
A diagnosis chart of nine species of fowl coccidia-Georgia Agri
cultural Experiment Stations - Dec. 1964, Technical Bulletin NS 39)
to know
O: no macroscopic lesions
1: rare sparse petechiae on the caecal mucosa
no thickening of the caecal walls - content
normal caecal
2: More lesions with blood in the contents
caecal - caecal wall somewhat thickened
presence of normal caecal content
3: significant amount of blood or pus caecal
Cecal wall strongly thickened - little or no
feces in the cecum
4: strongly distended caecal wall with clusters of
blood or caseous pus - non-existent faecal debris
or included in the heaps of dead animals
are rated 4.

- The indices of consumption (IC): ratio of the quantity of nour
ingested cheese from D-2 to D + 8 at weight gain from D-3 to D + 8.

BOARD

<tb> Groups
## EQU1 ## > C <SEP> T <SEP> M <SEP> L
<tb> Criteria
<tb><SEP> P <SEP> P <SEP> GP <SEP> P <SEP> GP <SEP> P <SEP> GP <SEP> P <SEP> GP
<tb> Weight <SEP> individual <SEP> 110 <SEP> 223 <SEP> 113 <SEP> 189 <SEP> 79 <SEP> 235 <SEP> 125 <SEP> 236 <SEP> 126
<tb>: <SEP> 111 <SEP>: <SEP> 261 <SEP> 150 <SEP>: <SEP> 212 <SEP> 101 <SEP>: <SEP> 235 <SEP> 124 <SEP>: <SEP > 228 <SEP> 117
<tb> 129 <SEP> 235 <SEP> 106 <SEP> 194 <SEP> 65 <SEP>. <SEP><SEP> 275 <SEP> 146 <SEP>. <SEP><SEP> 266 <SEP> 137
<tb>: <SEP> 138 <SEP>: <SEP> 312 <SEQ> 174 <SEP>: <SEQ> 181 <SEP> 63 <SEP>: <SEP> 245 <SEP> 107 <SEP>: <SEP > 200 <SEP> 62
<tb> Average <SEP> weight <SEP> 122 <SEP> 258 <SEP> 136 <SEP> 194 <SEP> 77 <SEP> 247 <SEP> 125 <SEQ> 232 <SEQ> 110
<tb> I <SEP> C <SEP> / <SEP> 1,902 <SEP> / <SEP> 2,987 <SEP> / <SEP> 2,018 <SEP> / <SEP> 2011
<tb>: <SEP> D + 5 <SEP>: <SEP> 0 <SEP> 2150 <SEP>: <SEP> 1580 <SEP>: <SEP> 1150
<tb><SEP> D + 6 <SEP> 0 <SEP> 1030 <SEP> 930 <SEP> 840
<tb> @@@ 103 <SEP><SEP> to <SEP> D + 7 <SEP> 0 <SEP> 420 <SEP> 22 <SEP> 73
<tb><SEP> TOTAL <SEP> 0 <SEP> 3600 <SEP> 2532 <SEP> 2063
<tb><SEP>%<SEP> / <SEP> 70.33 <SEP> 57.30
<tb>: <SEP> D + 5 <SEP>: <SEP> 0 <SEP> 4 <SEP>: <SEP> 2 <SEP>: <SEP><SEP> 3
<tb><SEP> M <SEP> to <SEP> D + 6 <SEP> 0 <SEP> 2 <SEP> 0 <SEP> 3
<tb> D + 7 <SEP> 0 <SEP> 1 <SEP> 0 <SEP> 0
<tb> D + 5 <SEP> 0 <SEP> 4 <SEP> 3 <SEP> 2
<tb> MF <SEP> to <SEP> D + 6 <SEP> 0 <SEP> 3 <SEP> 1 <SEP> 2
<tb> D + 7 <SEP> 0 <SEP> 1 <SEP> 1 <SEP> 0
<tb>: <SEP> 0 <SEP>: <SEP> 4 <SEP>: <SEP> O <SEP>: <SEP> 0 <SEP> 0 <SEP>
<tb> Lesions <SEP> 1 <SEP> 0 <SEP> 0 <SEP> 3 <SEP> 1
<tb> (number <SEP> of animals <SEP> 2 <SEP> 0 <SEP> 2 <SEP> 0 <SEP> 0
<tb> sore <SEP> by <SEP> note)
<tb><SEP> 3 <SEP> 0 <SEP> 0 <SEP> 1 <SEP> 0
<tb> 4 <SEP> 0 <SEP> 2 <SEP> 0 <SEP> 3
<tb> Mortality <SEP>% <SEP> 0 <SEP> 0 <SEP> 0 <SEP> 0
<Tb>
The results show that at a dose of 100 ppm / day, monensin lactone and monensin increase the weight gain of 15-day-old and E.tenella-infected cockerels compared to untreated contaminated animals when these products are distributed during 10 days. days.

 However, monensin lactone further inhibits oocystal excretion than monensin, which translates into a reduced risk of contamination of healthy animals with faeces from infested animals.

II. Toxicity
Acute toxicity tests were performed on male mice, orally and fasting for at least 6 hours. For this purpose, suspensions of the compound to be studied were used in water containing 10% gum arabic.

The animals were observed for 14 days and then slaughtered.

The following results were noted compared to monensin
LD50 (mg / kg)
Monensine lactone between 500 and 700
Monensine between 150 and 175
Under the same conditions, the toxicity of other polycyclic polyethers also appears much more important. The results below are from "The Merck Index", 10th edition (1983).

LD50 (mg / kg)
Lonomycin 50 Sal inomycin 50
Monensin lactone in the form of sodium salt is a known compound having been published, as well as its method of preparation in J. Am. Chem.

Soc. 1982, 104, pp. 7274-7281. This compound is presented as a synthetic intermediate without any pharmacological property or therapeutic application being reported
According to the process described, the sodium salt of monensin lactone is obtained by oxidizing at room temperature and in a methanol-ethanol mixture, the sodium salt of monensin using sodium metaperiodate.

Monensin lactone can be prepared at room temperature and in a water / tert-butanol mixture according to the method described in US Pat.
Antimicrobial Agents and Chemotherapy, 1967, pp. 359-362, that is, by oxidation of monensin by means of sodium metaperiodate.

 Generally, monensin lactone is obtained at ambient temperature and in an organic solvent, for example dioxane, by oxidation of monensin, for example by means of a metaperiodate such as sodium metaperiodate and acidification of the organic phase by means of a strong acid for example hydrochloric acid.

 As for the non-toxic salts of monensin lactone, these can be produced in a known manner by reaction of the monensin lactone with a suitable organic or inorganic base such as an alkali metal hydroxide, a metal-alkaline earth oxide, ammonia or an amine.

 Monensin lactone and its non-toxic salts can therefore be administered as medicinal active ingredient in the form of unit compositions suitable for administration in human or veterinary therapy or as a veterinary food additive.

 In its most general aspect and taking into account the final use chosen, the pharmaceutical or veterinary compositions in question are prepared by combining the active ingredient according to the invention with a suitable vehicle, diluent or excipient, the latter being selectable among substances such as distilled water, benzyl alcohol, lactose, starches, talc, magnesium stearate, polyvinylpyrrolidone, alginic acid, colloidal silica, microcrystalline cellulose, phthalic or acrylic esters , titanium dioxide, sweetening agents, etc.

 As regards the administration unit, this may take the form of, for example, a tablet, a dragee, a capsule, a capsule, a powder or a suspension. or a syrup for oral administration, a suppository for rectal administration, a sterile solution or suspension for parenteral administration.

 By way of example, an anticoccidial composition may be obtained by adding monensin lactone or a non-toxic salt thereof to the basic food ration in an amount of from 50 to 200 ppm by weight of the final mixture.

 Next, in a more particular but nonlimiting manner, a few embodiments of compositions according to the invention will be described.

 For example, powders for oral administration are prepared by simply grinding the active ingredient to a suitable fineness and mixing with a similarly ground diluent, which may be for example an edible carbohydrate compound, such as starch.

 Advantageously, a sweetening agent or a sugar and a flavoring oil are incorporated.

 Tablets may also be obtained by first forming a powder mixture from the active compound of formula I or a suitably ground salt thereof and a diluent or base such as starch, sucrose, kaolin, phosphate dicalcium etc ... then granulating or cutting after adding a lubricant and finally compressing. Granulation of the powder mixture can be done by wetting with a binder such as syrup, starch paste or acacia mucilage and forcing through a sieve.

 Another method of granulation is to cut the powder mixture by passing it through a tablet machine and breaking into pieces (sections) the resulting tablets imperfectly formed. The sections can be lubricated to prevent sticking by forming cubes by addition of a stearate salt, talc, or mineral oil. The lubricated mixture is then compressed to form the final tablets. If necessary, the tablets obtained may be sugar coated, coated with an enteric coating varnish or coated so that the active ingredient is released gradually.

 For veterinary use, the administration unit may take the form of a premix containing the active ingredient dispersed on the vehicle or diluent or the form of a food supplement containing the active ingredient mixed with the vehicle or diluent.

 In the latter two cases, such a vehicle is preferably a feed component for animals, for example oats, soybean seed, alfalfa, wheat, fermentation residues, crushed oyster shells, molasses, edible vegetable substances, soy flour, kaolin, talc, crushed limestone etc ...

 In the form of premix, compositions of the invention for combating coccidiosis may contain from 10% to 80% by weight of monensin lactone or a non-toxic salt thereof and from 90% to 20% of vehicle or diluent by weight of the composition. These premixes may be diluted with an animal feed supplement or may be added directly to an animal feed ration to provide a therapeutic diet that can be ingested by the poultry.

 Alternatively, monensin lactone and its non-toxic salts may be administered in the form of a solution or suspension containing a therapeutically effective amount of this active ingredient in the drinking water of poultry.

 Such a therapeutically active amount may vary from 0.5 mg to 30 mg per chicken per day. This daily dose may be, for example, 1.3 mg per 7-day-old chicken or 10.5 mg per 70-day-old chicken.

The following nonlimiting examples illustrate the preparation of the active ingredient according to the invention as well as compositions containing it.
EXAMPLE 1
Preparation of de (hydroxymethyl) -25 deoxy-25-oxo-25-monensin or monensin lactone

 In 7 ml of dioxane, 0.75 mM of monensin produced from strain S.cinnamonensis ATCC 15413 (US Patent No. 3,501,568) is dissolved and purified.

1.85 mM of sodium metaperiodate (NalO4) dissolved in 3 ml of water are then added and the solution is then placed under nitrogen and protected from light. Stirred for 2 hours with a magnetic stirrer and allowed to rest for about 15 hours. The mixture is diluted in approximately 200 ml of water and extracted 3 times with chloroform. The organic fractions are combined and acidified with 0.5N hydrochloric acid.

The acid phase is extracted with chloroform, the chloroform phases are combined and rinsed 3 times with deionized water.

 The rinsing waters are combined and extracted with chloroform. The organic phases are combined and dried over 4 hours over magnesium sulphate.

After filtration, the solution is evaporated with a rotary evaporator.

The product obtained is taken up in acetone and evaporated 3 times in order to dry well.

 In this way, de (hydroxymethyl) -25 deoxy-25-oxo-25-monensin is obtained.

Yield: about 100%
Purity: about 95%
Mp: 68-7O0C 25 43 (c = 16 mg / ml, chloroform) at 578 nm
Mass spectrum: FAB (fast atom bombing) technique
FAB -: M = 637 (deprotonated molecule: A)
FAB +: M = 661 (molecule plus sodium ion: AH Na +)
IR spectrum: 1720 cm 1 (much more intense line than monensin
indicating the presence of a C = O band).

NMR spectrum: The NMR spectrum confirms the structure of the molecule
disappearance of carbon 26 at 67.9 ppm and appearance of
carbon 25 at 174.5 ppm instead of 97.0 ppm.

EXAMPLE 2
Tablets containing monensin lactone, as an active ingredient, are prepared in association with an excipient to form the composition below:
mg
Monensine lactone 30
Lactose 173
Starch 25
Pregelatinized starch 20
Magnesium stearate 2
250
Mix thoroughly and for 15 minutes. in a mixing machine all the ingredients except the lubricant are kneaded by gradual addition of water. The mass is passed through a 1.25 mm sieve and the granulate is then dried in a forced ventilation oven until a relatively low residual moisture (about 2%) is obtained. The granule is uniformized, the lubricant is added and compressed tablets are formed.

EXAMPLE 3
Tablets containing monensin lactone, tablets having the following composition are prepared
mg
Monensine lactone 30
Lactose 192
Microcrystalline cellulose 45
Starch 30
Magnesium stearate 3
300
The powders are passed through a 0.3 mm sieve and then the monensin lactone is mixed with the same amount of lactose. The microcrystalline cellulose, the starch and the lactose residue are then added and mixed again. The magnesium stearate is then added and the mixture is mixed until a homogeneous unit is obtained which is then compressed and granulated.

 The granules thus obtained are used to form tablets by compression.

EXAMPLE 4
Capsules containing the following composition mixture are prepared
mg
Monensine lactone, sodium salt 60
Lactose 194
Starch 30
Talc 15
Magnesium stearate 1
300
The sodium salt of the monensin lactone, the lactose and the starch are intimately mixed in a mixer and then the talc and the magnesium stearate are added. The mixture is rapidly mixed and then introduced into soft capsules of gelatin.

EXAMPLE 5
Feed premixes are prepared by intimately mixing the following ingredients:
Ingredients Parts by weight 1) Monensine lactone 50
Alfalfa granules 50 2) Monensine lactone 40
Soluble Molasses 60 3) Monensine Lactone 30
Dried grains of corn 70 4) Sodium salt of monensin lactone 40
Corn flour 30
Dried grains of corn 30) Sodium salt of monensin lactone 60
Ground soybean seeds 40 6) Monensine lactone 20 sodium salt
Dried grains of mals 60
Wheat bran 20 7) Monensine lactone 80
Wheat bran 20 8) Monensine lactone 25
Soybeans 75 9) Monensine lactone 15
Bark of rice grain 85

Claims (6)

 25-deoxy-25-oxo-25-monensin of formula  that they contain, as essential active ingredient, the de (hydroxy 1. Pharmaceutical or veterinary compositions characterized in that  diluent or excipient.  or a non-toxic salt salts in combination with a vehicle, 2. Veterinary compositions with anticoccidial activity, carac  contained in that they contain as essential active ingredient,  de (hydroxymethyl) -25-deoxy-25-oxo-25-monensin or a salt thereof  non-toxic in association with a vehicle, diluent or excipient  appropriate. 3. Compositions according to one of claims Lou 2 characterized in that  that they are in the form of a dosage unit. 4. Compositions according to one of claims 1 or 2 caractérizéesen ce  they are in the form of premix. 5. Compositions according to claim 2, characterized in that they are  in the form of a food ration containing the active ingredient  from 50 to 200 ppm by weight of the final mixture. 6. Compositions according to claim 4, characterized in that  contain from 10% to 80% by weight of active ingredient.