DK2349272T3 - Veterinær anti-prolaktin-sammensætning til drøvtyggere - Google Patents

Description

Anti-prolactinic veterinary composition for ruminants

The object of the invention is an anti-prolactin veterinary composition for administration to ruminants.

This composition includes cabergoline as an anti-prolactinic dopamine receptor agonist compound, and is particularly useful in promoting breast involution, breast tissue regeneration and lactation suppression conditions. The compositions according to the present invention are also administered during the gestation period because they do not have a deleterious or abortive effect on the gestation of ruminants.

Lactation generally lasts from 5 to 20 months in ruminants, and averages 10 months in dairy cows. After this lactation period, milking is usually stopped abruptly, and the cow is rested for a period that traditionally lasts 60 days (called the dry period) until calving, the starting point of the next lactation. In dairy farms, it is recognized that this dry period significantly improves the health and well-being of ruminants. This physiological resting of the udder is necessary for the regeneration of the mammary gland and the preparation for future lactations. It also limits the occurrence of health problems when milking stops and related to calving, such as the appearance of udder inflammation and infection, which have a significant economic impact.

Anti-prolactinic compounds are prescribed in human medicine for the treatment of hyperprolactinemias, pituitary adenomas, Parkinson's disease, or to cause lactation suppression in some women who cannot breastfeed for medical reasons. Such drugs contain as active ingredients inhibitors of the pituitary secretion of prolactin, a hormone that activates milk secretion. These include, for example, drugs such as Parlodel® (bromocriptine, Novartis), Dostinex® or Cabaser® (cabergoline, Pfizer), for which it is explicitly stated that they may be taken under any circumstances during pregnancy due to their harmful effects on the foetus. It is recommended that prescriptions for lactation suppression are administered immediately after delivery in order to stop milk production within 24 hours.

In veterinary medicine, compositions based on a prolactin inhibitor such as Galastop® (cabergoline, Ceva Animal Health) can also be administered as treatments for pseudo-gestation lactations in cats or bitches.

They are also prescribed following early weaning, abortions, or when the litter is immediately withdrawn after parturition. In animals, as in human patients, prolactin secretion inhibitors have always been prescribed outside gestation or pregnancy due to toxicity effects, risks of foetal malformations, and risks of spontaneous abortion.

In ruminant mammals, prolactin is known to play an important role in the maturing of the mammary gland and the onset of lactation after calving but has no effect on milk production during established lactation (Karg H. & Shams D., J. Reprod Fertil., 1974 Aug ; 39(2) ;463-72 — Shams et al., Experentia, 1972 ; (28) :697-699 — Akers et al., Endocrinology, 1981; (109) :23-30 — Plant K. et al., Domest Anim Endocrinol, 1987 ; (4); 279-290 — Knight, Livestock Production Science, 2001, (70); 87-93 - Dalh et al, J ;Anim. Sci., 2003 ; (81), supl3 ; 11-17); G. Leitner et al., Livestock Science 110 (2007) 292-297 describe the use of casein hydrolysates to improve the well-being of dairy cows at the time of milking cessation.

Unlike many of these publications which teach that the decrease in prolactin has no or very little effect on milk production during established lactation in the main dairy species, such as cows and goats, the Applicant surprisingly discovered that the single administration of a dose of an anti-prolactin dopamine receptor agonist compound such as cabergoline, triggered a substantial decrease in lactation and effectively induced the initial suppression phase, which consists of breast involution.

Similarly and against all expectations, the Applicant discovered that veterinary compositions containing anti-prolactinic dopamine receptor agonist compounds could be administered to ruminants in order to induce decreased lactation, promote breast involution and tissue suppression conditions, even during gestation without causing deleterious or abortive effects.

The administration of these veterinary compositions to ruminants has proved to be particularly beneficial since it has significantly reduced lactation and promoted breast involution, even during gestation, resulting in improved suppression conditions. The veterinary compositions according to the invention also ensure a significant improvement in udder health, a better regeneration of damaged secretory tissues in the udder, as well as a reduction in the frequency of udder diseases and/or infections or ruminant mastitis during subsequent lactations.

Summary of the invention

This invention concerns veterinary compositions containing cabergoline administered to pregnant ruminants for use in inducing lactation suppression and promoting breast involution in a manner that promotes their well-being or in treating and/or preventing diseases and/or infections of their udders.

According to the invention, these compositions are administered in effective therapeutic quantities, in particular as a single treatment, so as to induce a decrease in lactation. Also, these compositions can even be administered to pregnant ruminants without causing a deleterious or abortive effect on pregnancy. They thus significantly promote the well-being of ruminants, and are particularly useful in mammary involution, treatment and/or prevention of inflammation and infection of ruminant udders. The object of the invention is also to develop methods to promote breast involution in ruminants, as well as methods to prevent and/or treat infections of ruminant udders such as mastitis. Finally, the object of the invention is to provide kits to promote breast involution and the reduction of milk production in ruminants, as well as kits for the treatment and/or prevention of mastitis in ruminants.

Brief description of the drawings

Fig· 1: is a graph showing the percentage change in milk production in the morning and evening between Day D-7 and Day D7 (compared to a baseline of 100) following a single injection of 5mcg/kg cabergoline on Day DO, in a placebo group and in a group of treated animals (C646).

Fig. 2 : is a graph that represents the percentage change in prolactin levels observed every 24 hours during period I (D-2 and D-l) and period II (between DO and D7), compared to a baseline of 100, following a single injection of 5mcg/kg cabergoline on day DO, in a placebo group and in a treated animal group (C646).

Detailed description of the invention

The object of the invention is to provide a veterinary composition comprising cabergoline administered in a single dose to induce suppression and a substantial decrease in lactation in the treated animal from the first and subsequent milking. The object of the invention is to use the said veterinary composition administered in a sufficient therapeutic quantity and in a single dose to induce a substantial decrease in lactation.

Prolactin is a pituitary hormone that has a mammotropic and lactogenic effect, i. e. it activates the growth of the mammary glands and milk secretion. The release of prolactin is under the stimulative influence of prolactoliberin and the inhibitive influence of dopamine. The inhibitory action of dopamine in the pituitary gland is mediated by post-synaptic dopamine receptors, such as the D2 receptor in particular.

The compounds are termed anti-prolactinic when they inhibit the release of prolactin. These may be dopaminergic or serotoninergic compounds. Dopaminergic compounds are dopamine receptor agonists capable of binding to dopamine receptors present particularly in the prolactin secreting cells of the anterior pituitary gland, to inhibit prolactin secretion.

Preferably, these dopaminergic anti-prolactin compounds bind to dopamine receptors. Even more preferably, these compounds bind specifically to dopamine D2 receptors. Anti-serotoninergic anti-prolactinic compounds act by stimulating the release of dopamine into the hypothalamus, thereby inhibiting the secretion of prolactin. The veterinary compositions according to the present invention include cabergoline as a dopaminergic anti-prolactinic compound.

Cabergoline, whose chemical name is N-[3-(Dimethylamino)propyl]-N-[(ethylamino)carbonylj- 6-(2-propenyl)-8g-ergoline-8-carboxamide, is an agonist antiprolactin compound specific for dopamine D2 receptors. It is described in particular in US Patent 4,526,892. Its structural formula is as follows:

Cabergoline is the active ingredient in human medicines marketed under the names Dostinex® and Cabaser®. It is also the basic active principle of veterinary

compositions marketed under the name Galastop® for cats or bitches subject to pseudogestation lactations. Whether it is Dostinex® or Galastop®, these cabergoline compositions are never administered during pregnancy or gestation.

Until now, cabergoline has always been administered outside pregnancy or gestation.

However, the Applicant found surprisingly that this compound, when administered to ruminants, in single administration and in effective therapeutic amounts, actually induced a significant decrease in lactation, to promote breast involution and improve lactation suppression. Contrary to what has been known until now, this composition has proved to be particularly beneficial since it allowed de facto production to be effectively suppressed, particularly during ruminant gestation, without causing lethal and abortive effects. Finally, the compound according to the invention ensured a significant reduction in the risk of mastitis in ruminants thus treated.

As demonstrated in the examples below, a single dose may be administered to non-human animals to promote breast involution and lactation suppression conditions, whether during pregnancy or after calving. Effective therapeutic quantities or doses may vary depending on the ruminants to be treated and the method of administration of the compositions. The dosages can easily be determined by systematic tests based on the examples below within the reach of the skilled person. For example, the effective therapeutic doses according to the present invention are between 5 and 50mcg/kg, or between 5 and 25 mcg/kg.

Preferably according to the invention, the compositions are administered in a single dose per treatment and per animal so as to obtain a substantial reduction in lactation of between 5 and 60%, between 20 and 50%, or between 25 and 35%, over a period of time ranging from 1 day, 2 days, 3 days, 4 days, 5 days, 6 days and up to 7 days after administration of the composition.

As shown in Example 2, the decrease in lactation is significant from the first milking, but also from the next milking.

It is possible to administer the compositions according to the invention to pregnant ruminants and more precisely during the last two thirds of gestation, without however causing deleterious or abortive effects. For example, the said veterinary compositions according to the invention can be administered from the 3rd month and more after the gestation of the dairy cows. Preferably, they may be administered to pregnant ruminants between the 3rd month and the 8th month of gestation.

According to the invention, ruminants are defined as herbivorous mammals such as cattle, sheep, goats, camelids or bovids. The compositions according to the invention are administered to ruminant mammals producing milk, such as preferably dairy cows and ewes.

According to the invention, the compositions can be administered during the gestation period or outside the gestation period. They promote the reduction of lactation and act on the succession of hormonal, morphological and physiological upheavals which affect in particular the udder during suppression, and in particular from the initial phase of suppression, breast involution, during which the secretory tissue breaks down and atrophies. Surprisingly, no deleterious effects of toxicity, foetal malformations or abortive effects were observed when administered during ruminant gestation.

Veterinary compositions may be administered by any route of administration well known in the field and adapted to the treatment of each animal. Preferably, they are administered by the dermal, oral and parenteral routes. Even more preferably, they are administered by parenterally, including through intramuscular or subcutaneous injection. They are then presented in all appropriate forms according to the desired modes of administration. They can therefore be in the form of an oral or injectable liquid solution or suspension, or in solid or semi-solid form, powders, tablets, capsules, granules, tablets, capsules, sprays, tablets, pills, tablets, or pastes.

Advantageously, the veterinary composition will be administered in a single injectable dose.

Depending on the formulations of the compositions and drugs used, they may also include ingredients conventionally used in pharmacy for the preparation of liquid or solid formulations for oral or parenteral administration. In addition, in the case of oral formulations, they can be administered directly to ruminants or mixed with feed.

Also, compositions according to the invention may include, depending on the type of formulation, a solvent, a flowing agent, a lubricant and any excipient of suitable mass, such as lactose, cellulose or starches. Stearic acid, magnesium stearate, L-leucine or, for example, glycerol tri behenate can be used as a lubricant. Sodium carboxymethylamidone, cross-linked sodium carboxymethylcellulose or cross-linked polyvinylpyrrolidone can be used as a disintegrating agent. Pure silica or colloidal silicon dioxide can be used as a flowing agent. Solid oral forms can be in the form of coated tablets.

Injectable preparations are prepared by mixing effective therapeutic amounts of at least one anti-prolactin compound as previously described with a solvent, pH regulator, buffering agent, suspending agent, solubilizing agent, stabilizer, tonicity agent and/or preservative, and converting the mixture into a subcutaneous or intramuscular injection by a conventional method. Solvents include oily solvents such as medium chain 8-10 triglycerides or a mixture of capric acid, caprylic acid and triglycerides, such as those marketed under the name Mygliol®812. If necessary, the injectable preparations can be lyophilized using a conventional process. Examples of suspending agents include methylcellulose, polysorbate 80, hydroxyethylcellulose, xanthan gum, sodium carboxymethycellulose and polyethoxylated sorbitan monolaurate. Examples of solubilizing agents include castor oil solidified with polyoxyethylene, polysorbate 80, nicotinamide, polyethoxylated sorbitan monolaurate, macrogol and castor oil fatty acid ethyl ester. In addition, stabilisers include sodium sulphite, sodium metalsulphite and ether, while the preservatives include methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, sorbic acid, benzyl alcohol, phenol, cresol and chlorocresol. Mannitol is an example of a tonicity agent. When preparing injectable solutions or suspensions, it is desirable to ensure that they are isotonic to blood.

Advantageously, the veterinary compositions or drugs according to the invention can be administered in combination with standard treatments for ruminant mastitis. Examples of standard treatments or prophylactic compositions for mastitis include local teat disinfectant compositions, antibiotics such as group M penicillins, cephalosporins, gentamycin or colistin, or enzymes such as lysozyme or muramidase.

The object of the invention is also to use effective therapeutic quantities of at least one anti-prolactinic dopamine receptor agonist compound as previously described for the preparation of veterinary compositions or drugs for administration to ruminants during or outside the gestation period in order to improve their well-being and health, to promote the suppression of milk production in particular, to promote breast involution, and to prevent and/or treat udder inflammation or infection. When veterinary compositions or drugs are administered during the gestation period at therapeutic doses compatible with maintaining gestation, they do not have deleterious or abortive effects.

The present invention also concerns methods for suppressing milk production and methods for the treatment and/or prevention of mastitis including dermal, oral or parenteral administration of previously described veterinary compositions to ruminants in effective therapeutic amounts. The methods according to the present invention are particularly advantageous since they promote breast involution and the suppression of lactation of ruminants, even pregnant ruminants, without causing deleterious and abortive effects.

The object of the invention is also to provide a veterinary kit to help reduce the production of ruminant milk, improve the well-being and health of these ruminants, promote breast involution, treat and/or prevent udder diseases, in particular mastitis. The kits according to this invention comprise at least one compartment for possibly sterile packaging comprising an effective therapeutic amount of cabergoline as an anti-prolactinic dopamine receptor agonist compound as previously described for administration to ruminants. The kit includes the means for administering the compositions by dermal, oral or parenteral route as well as an instruction sheet on how to administer the compositions or veterinary drugs according to the invention.

EXAMPLES

Example 1: Preparation of veterinary compositions with a cabergoline base

Preparation of 9 litres of a preparation for injectable solution containing 500 mcg/ml of cabergoline.

Formula:

Active principle: 4.53 g of cabergoline (titrating 100%)

Excipient: triglycerides of a medium chain q. s. 9 I.

Step 1: 4.53 g of cabergoline and 1.5 kg of medium chain triglycerides were measured in a container of adequate capacity, then the mixture was stirred using a magnetic stirrer (500 rpm) for at least 60 minutes for complete dissolution.

Step 2: In a dry container, 5 kg of medium chain triglycerides were measured, the nitrogen flow and mixer were started, and then the cabergoline in concentrated solution, obtained in step 1, was added to the container. Agitation was maintained for 30 minutes and then the volume was increased to the final volume of 9 litres by adding medium chain triglycerides. Agitation was maintained for another 30 minutes and then the solution was filtered under pressurised nitrogen through a 0.45 micron calibrated cartridge. The filtrate was collected in a suitable container previously disinfected with steam and dried with a stream of nitrogen. The resulting solution was sterilized at 121°C for 15 minutes, and then distributed into sterile and pyrogen-free vials closed with rubber stoppers and aluminium capsules, washed and autoclaved.

Example 2 : Clinical study of the efficacy of cabergoline-based veterinary compositions in ruminants

The objective of this study was to evaluate the decrease in milk production in 8 multiparous cows of the Prim'Holstein breed in lactation for at least 6 months. These cows are 3 to 9 years old and have a production level varying from 20 to 31 kg per day at the start of the study. The trial was organized as a crossover study (2 treatments x 2 periods). The formulation described in Example 1, containing 500 mcg/ml of cabergoline in an injectable solution, was used in this study.

Test Treatment (C646): 500 mcg/ml cabergoline injectable solution, 1 single intramuscular injection at a dose of 5 mcg/kg or 1 ml per 100 kg body weight.

Control treatment (Placebo): water for injectable preparation, 1 intramuscular injection at the same volume as the test treatment, i.e. 1 ml per 100 kg.

The treatment sequence was randomised (A-B or B-A) with a 3-week wash-out period between the 2 treatments. Individual milk production was monitored at each milking over a period ranging from 7 days before treatment to 7 days after treatment. Blood samples were taken daily to monitor changes in plasma prolactin levels over a period ranging from 2 days before treatment to 7 days after treatment.

The results shown in Fig. 1 show that injecting the Test treatment at a dose of 5 mcg/kg resulted in a reduction in milk production:

From the first milking after the injection of the Test treatment (12 hours), the reduction in milk production was 27% on average, ranging up to 42% depending on the individual.

Over the follow-up period of 7 days after Test treatment, the overall reduction in milk production was 15% on average, ranging up to 27% depending on the individual.

By generalising the results to a cow population, these results make it possible to estimate that the reduction in milk production from the first milking varies between 5 and 60% depending on the individual. The reduction in milk production after injection of the Test treatment is likely to be between 25 and 35% at the next milking. This decrease was maintained for approximately 2 days. A gradual recovery in milk production was then observed, but without returning to its initial level after 1 week.

At the same time, plasma prolactin concentrations decreased as shown in Fig. 2. The decrease in plasma prolactin concentration was rapid, as early as 24 hours after injection, and was still present one week after treatment. This decrease in prolactin varies from 10 to 80% depending on the individual. The single administration of cabergoline demonstrated the correlation between the significant decrease in milk production and the decrease in plasma prolactin.

Example 3: Clinical safety study in ruminants of cabergoline-based veterinary compositions

This study aims to show the absence of abortive risk and foeto-toxicity when using cabergoline in pregnant ruminants, even in the case of an overdose of the veterinary composition based on carbergoline compared to the dosage prescribed according to this invention. Groups of cows at different stages of gestation received treatment at different doses: 3 months gestation (6 animals), dose of 15 mcg/kg - 2 injections at 48 hours apart; 7 to 8 months of gestation: (5 animals), 15 mcg/kg dose - 2 injections at 48 hours apart; 6.5 to 7.5 months of gestation: (11 animals), 25 mcg/kg dose - 2 injections at 48 hours apart.

The formulation described in Example 1, containing 500 mcg/ml of cabergoline solution for injection, was used in this study.

Regular clinical examinations were performed over a period ranging from 7 days before treatment to 15 days after treatment:

Daily general clinical and abortion monitoring;

Clinical examination of the genital tract including transrectal palpation and ultrasound to assess foetal distress and viability.

Blood samples were taken 1 day before and 15 days after treatment to monitor plasma concentrations of different hormones and proteins that indicate placental and foetal health, such as progesterone, estrone sulphate, and PAG (Pregnancy Associated Glycoprotein). Animals were monitored until calving to assess the health status of newborn calves. The results show that two injections of cabergoline at 48-hour intervals up to the dose of 25 mcg/kg do not cause abortion or any adverse effect on the gestation, viability and health of new-born calves. No variation in hormones indicative of placental or foetal toxicity in relation to treatments at different doses was observed.

In addition, the animals generally tolerated the treatments well. In conclusion, a single or repeated administration of cabergoline at 48 hours up to at least 25 mcg/kg is safe in pregnant cows.

Example 4 (reference example)

Clinical study of the efficacy and safety of abortion in ruminants of quinagolide-based veterinary compositions: 9 pregnant Prim'Holstein multiparous cows (about 600 kg) of about 7 months of age with a milk production level of about 18 kg were included in the study. The study was organized as a crossover study: 2 treatments x 2 periods, with a 3-week wash-out period.

The animals received the test treatment intramuscularly from adapted injectable formulations.

Test Treatment: injectable solution of 0.5 mg/ml quinagolide, 1 single intramuscular injection at a dose of 1 to 3 mg/cow or 1 ml per 100 kg body weight.

Control treatment (Placebo): water for injectable preparation, 1 intramuscular injection at the same volume as the test treatment, i.e. 1 ml per 100 kg.

Milk production monitoring was organised 7 days before and after the day of treatment for each period, and abortion monitoring similar to that described in example 3 is performed throughout the study.

Example 5 (reference example)

Clinical study of the efficacy and abortion-safety in ruminants of bromocriptine-based veterinary compositions: 9 pregnant Prim'Holstein multiparous cows (about 600 kg) of about 7 months of age with a milk production level of about 18 kg were included in the study. The study was organised as a crossover study: 2 treatments x 2 periods, with a 3-week "wash-out" period.

The animals received the test treatments intramuscularly from adapted injectable formulations.

Test treatment (bromocriptine): 10 mg/ml injectable solution of bromocriptine, 1 single intramuscular injection at a dose of 100 to 200 mcg/kg or 1 ml per 100 to 50 kg body weight.

Control treatment (Placebo): water for injectable preparation, 1 intramuscular injection at the same volume as the test treatment, i.e. 1 ml per 100 kg.

Milk production monitoring was organised 7 days before and after the day of treatment for each period, and abortion monitoring similar to that described in example 3 was carried out throughout the study.

Claims (10)

1. Veterinær sammensætning omfattende cabergolin til anvendelse til at inducere depletering af laktation og fremme yver-involution hos drægtige drøvtyggere for at forbedre deres velfærd eller at behandle og/eller forebygge sygdomme og/eller infektioner i deres yvere.

2. Sammensætning til anvendelse ifølge krav 1, kendetegnet ved, at sammensætningen indgives i en effektiv mængde til at inducere depletering eller reduktion af laktation omfattet mellem 5 og 60%, 20 og 50%, eller mellem 25 og 35%.

3. Sammensætning til anvendelse ifølge et hvilket som helst af kravene 1 til 2, kendetegnet ved, at sammensætningen indgives i en effektiv mængde uden at forårsage nogen skadelige eller abortive virkninger på de drægtige drøvtyggere.

4. Sammensætning til anvendelse ifølge et hvilket som helst af kravene 1 til 3, kendetegnet ved, at sammensætningen indgives via parenteral, kutan eller oral vej.

5. Sammensætning til anvendelse ifølge et hvilket som helst af kravene 1 til 4, kendetegnet ved, at sammensætningen indgives via injektion, især som en enkelt indgivelse via intramuskulær injektion, eller via subkutan injektion.

6. Sammensætning til anvendelse ifølge et hvilket som helst af kravene 1 til 5, kendetegnet ved, at drøvtyggerne er planteædende pattedyr valgt fra kvæg, får, geder, kameler eller horndyr.

7. Sammensætning til en anvendelse ifølge et hvilket som helst af kravene 1 til 6, kendetegnet ved, at sammensætningen indgives i en enkelt effektiv dosis per drøvtygger og per behandling.

8. Sammensætning til anvendelse ifølge et hvilket som helst af kravene 1 til 7, kendetegnet ved, at reduktionen af laktation eller depletering af laktation opnås op til 7 dage efter behandlingen med sammensætningen eller forekommer ved den første malkning og næste malkning af de behandlede drøvtyggere.

9. Kit til veterinær anvendelse omfattende en veterinær sammensætning omfattende cabergolin, til anvendelse til at forbedre velfærden, til at fremme yver-involution eller til at behandle og/eller forebygge forekomster forbundet med yver-sygdomme hos drægtige drøvtyggere.

10. Kit til anvendelse ifølge krav 9, til behandling og/eller forebyggelse af yver-betændelse hos drægtige drøvtyggere.