KR20010080571A - Triazineone compounds for treating diseases due to sarcosystis, neospora and toxoplasma - Google Patents

Abstract

The present invention is a parasitic neurologic or infected animals represent the animal or its susceptibility infected suffering from balyukbu conductive disorders, such as at the time of sarcoidosis, which may be treated with triazine-on compound styryl Dia (Sarcocystidiae) or toxoplasmosis (Toxoplasmosis) A method of treating said animal therapeutically or metadefensively, comprising administering a pharmaceutically effective amount of a compound, including treating at a single high dose.

Description

Triazineone compounds for treating diseases due to sarcosystis, neospora and toxoplasma}

The present invention relates to triazineone compounds for treating animals infected with parasites that cause abortigenic or neurologic diseases. More specifically, the present invention relates to triazineone compounds useful for treating parasitic protozoa, such as coccidia , causing dysplasia or neurological disease.

Triazinedione compounds such as triazinedione, for example diclazuryl compounds, and triazinetrione, for example tortrazilyl compounds, treat and protect a variety of mammals, insects and fish from diseases caused by a wide range of protozoa. (US Pat. Nos. 4,933,341; 4,935,423; 5,114,938; 5,141,938; 5,188,832; 5,196,562; 5,256,631 and 5,464,837). Protozoans susceptible to such compounds include parasites that infect the intestines of birds, mammals, and insects, causing diarrhea, weakness, nausea, and vomiting. In general, the mode of action of triazineone is to attack the intermediate parasitic stages found in the intestinal and intestinal wall cells to swell the parasitic reticulum, the nuclear space and the mitochondria. This is known to interfere with the fissile capacity leading to shizont and microgamont, so that only a small amount of merozoit and reproductive system are formed, respectively. The end result was that the latter phase of the parasite lost its ability to penetrate into new mammalian cells, virtually stopping replication of the parasite in the host.

Of particular interest here are certain protozoa which have been considered to cause neurological and / or dysplasia in animals since the 1970s. Successful isolation of some of these protozoa and culturing in vitro proved difficult. For example, they did not successfully separate from the brain or cerebrospinal fluid until the end of 1980. Effective anti-protozoal agents that can cross the blood-brain and placenta barriers without harmful side effects, as neurological diseases can be caused by parasites that infect the brain and dysplasia has been found to infect the fetus. Is desired. Agents that can cross the animal's blood-brain gate or placental gate are very rare. However, many agents known in the art that can effectively treat parasitic infections of the brain through the blood-brain barrier and / or placental barrier have adverse side effects and therefore cannot be used without great risk. Thus, no drug has been approved as an effective therapeutic agent for the neurological or dysplastic disease. The parasite disease will then be briefly described.

Equine protozoan encephalomyelitis (EPM) is a neurological disorder in horses that is particularly associated with stressed colts (e.g., horses with racehorses and horses with hematoma), which have a severe financial impact on the horse industry. Disease. EPM was first recognized as a disease in the 1970s, was not cultured from horses with EPM, and until 1991 was called Sarcocystis neurona . In 1997, Neospora spp . (Now named Neospora hugesi ) was isolated from the brains of horses with EPM. Thus, it has now been suggested that EPM can be caused by such newly recognized organisms alone, by Sarcocytis neurons alone or by a combination of the two. EPM most often causes asymmetry, ataxia, weakness and stiffness. These diseases can mimic almost all neurological symptoms. This can occur with hyper- or chronic symptoms. Chronic forms are often latent onset and difficult to diagnose until the end of the disease process, resulting in death. If the symptoms are mild, the only clinical signs may be unclear pelvic limb slack or weak breathing difficulties. In the most severe cases, the horse cannot swallow or stand. Currently, in the most severe cases, it is known that parasites such as Sarcocytis neurona infect the brain and significantly damage it. The clinical signs of EPM are caused by infiltration of inflammatory cells, edema and neuronal death in connection with mesozoites and meronts of the central nervous system (CNS) as well as by direct damage of neurons (brain and spinal cord) by parasites. Caused by brain injury. At present, there are no approved therapeutic or prophylactic agents to rescue EPM. The pharmaceutical trimethoprim-sulfonamide combination is being used. However, these therapeutic agents are expensive and require significant repeated administration.

Another coccidia parasite, Toxoplasma gondii , was later known and isolated from the intestine and muscle tissue of the first cat. The final host for this parasite is cats, which the organism can linger for a long time, during which oocytes spread to other animals, including cattle, sheep, pigs, and humans. Infection of sheep, cattle and humans involves dysplasia and congenital diseases, mainly affecting the central nervous system. It has also recently been accompanied by dysplasia and malformations in young cats born to infected female cats who were seronegative before infection during pregnancy. Hosts other than cats, such as cattle, sheep, pigs, and humans, do not produce oocytes but develop them, thus tachyzoite and bra, causing clinical dysfunction-neurological symptoms and dysplasia by fetal defects. Desserts ( bradyzoite ) can suffer from invading muscles and brain. Sixty percent of cats were reported to be serologically positive in Toxoplasma gondie. Once again, there is no approved therapeutic or prophylactic agent for toxoplasmosis.

Another coccidia parasite, Neospora caninum , causes both neurological and dysplastic diseases in animals. It was first isolated from dogs in 1988. This was previously confused with Toxoplasma condy. Diseases caused by these parasites occur most severely in transplacentally infected puppies and are characterized by progressive ascending paralysis of puppies, especially puppies hind limbs; Multiple myositis and hepatitis can also occur. This disease is more recently recognized as a major cause of dysplasia and neurologically related leg defects in newly born calves. Microscopic lesions of non-purulent encephalitis and myocarditis in dysplastic fetuses can be found in the brain, spinal cord and heart. Recently, the final host for neospora caninum has been identified as an individual. In this case too, there is no approval as a therapeutic or prophylactic agent for neosphora caninum and horse neospora resting in horses.

References known in the art, including the references cited above, are directed to treating animals infected with coccidia or, more specifically, coccidia of the Sarcocystidae family that cause dysplastic or neurological diseases without undue side effects. No suggestion or disclosure has been made for the use of triazineone compounds, such as tortrazilyl or toltrazuryl sulfone (recently renamed "ponazilyl"). Therefore, it is desirable to treat animals suffering from parasitic diseases characterized by neurological or dysplasia in a safe and improved manner.

As described above, the present invention is characterized in that a pharmaceutically effective amount of a triazineone compound is administered to an animal suffering from parasitic neurological or dysplasia that exhibits sensitivity when treated with the triazineone compound. A method of therapeutically treating an animal, provided that the compound is not diclazilyl or toltrazilyl if the disease is sarcocistis neurona. As used herein, the term “pharmaceutically effective amount” refers to in vivo or in vitro the growth of coccidia in which the amount of triazineone administered causes parasitic protozoa, typically neurological disease and / or dysplasia. It is high enough to contain it. Pharmaceutically effective amounts control parasites in infected tissues, thus enhancing the health of the animal.

The present invention also encompasses a method for metaphylactically treating an animal infected with a parasite that can cause parasitic neurological or dysplasia that exhibits sensitivity when treated with a triazineone compound. Metaprotective treatment is characterized by administering the triazineone compound to the animal using a metaprotective effective regimen. The term "metaprotective effective regimen" means administration of a set intermittent dose of the triazineone compound for a long time until the animal exhibits a protective immune response or exterminates the parasite by killing the parasite. Typically, the regimen effectively kills parasites and prevents clinical symptoms of the disease. In particular, where parasites are difficult to control, metaprotective effective doses may also be administered for prolonged periods of up to 5 years or for the lifetime of the animal. In metaprotective treatment, preferred triazineone compounds are triazinetriones including, but not limited to, tortrazilyl and ponazilyl.

In addition, the present invention encompasses the treatment of animals in a single high dose. The method comprises a single dose of a pharmaceutically effective amount of a triazineone compound in a high dose to an animal suffering from a parasitic neurological or dysplastic disease that is susceptible to treatment with triazineone. The term "single high dose" means the amount administered only once. This amount is significantly higher than the dose used for treatment or metaprotective treatment; Effective in controlling parasitic diseases causing disease; By itself it does not cause adverse effects such as toxicity. Thus, a single high dose of triazineone is greater than 10 mg / kg. These and other aspects of the invention are described in further detail below.

As mentioned above, the present invention provides a pharmaceutically effective amount of triazine in an infected or diseased animal suffering from a parasitic disease characterized by neurological or dysplasia that is sensitive to treatment with a triazineone compound. And a method for treating an infected or diseased animal suffering from said disease, characterized by administering a warm compound. Examples of animals include, but are not limited to, horses, cattle, cats, dogs, pigs, sheep, birds, insects, and humans. Parasites that infect or cause disease are the coccidia of the sarcosididae family, which can appear as neurological or dysplastic diseases. Examples thereof may be selected from, but are not limited to, the group consisting of Sarcosistis species, Neospora species and Toxoplasma species. Sarcosistida may be selected from, but is not limited to, a group consisting of Sarcossis neurona, Neospora resting, Neospora caninum and toxoplasma gondi. Protozoan infections or diseases include, but are not limited to, EPM, neoporosis and toxoplasmosis.

In the practice of the present invention, symptoms of neurological and dysplasia may be alleviated by treating parasitic infections or diseases caused by the protozoa described herein. In general, these symptoms include limping, ataxia, paralysis, dysplasia, fragile newborns and other related diseases. For therapeutic treatment, animals that already exhibit symptoms of the disease are treated with triazineone compounds. Typically, the treatment period is about 28 to 90 days and preferably about 28 to 60 days. For therapeutic treatment, the treatment regimen may be once a day, at least twice a day, once every other day or once a week, depending on factors such as the severity of the disease and disease-induced parasites. In some cases, however, the treatment regimen can sometimes last indefinitely for the life of the animal. The latter treatment is necessary if the animal is infected with parasites of a more resistant species. However, this treatment can be prolonged for as long as necessary until no symptoms of the disease are seen. Preferred treatment is once daily for about 28 days.

In the case of metaprotective treatment, infected animals are treated to protect these animals from the clinical characteristics of the disease. Such treatment eventually gives the animal the ability to control parasites by establishing an effective immune response to protect against future infection without the need for additional administration of triazineone compounds. Metaprotective activity according to the present invention means the use of triazineone compounds as established intermittent therapeutic regimens (metaprotective effective regimens) to control protozoa which can infect animals after conventional treatment. Thus, metaprotective effective regimens are administered to reduce their likelihood of causing disease by killing protozoa or reducing their number. In essence, a metadefensively effective regimen may be administered about once a month over the life of the animal or until a unique elimination mechanism, for example an effective immune response, is developed in the animal and protected from future infection. The latter can happen within five years. As can be seen, metaprotective treatment involves neurological symptoms or dysplasia when animals are infected with the protozoa described herein until they have passed a significant time (eg 2 to 6 months after infection). It is based on the recognition that it does not exhibit clinical symptoms such as In contrast, enteroprotozoan infections express themselves shortly after infection. According to the present invention, metaprotective treatment prevents the parasite from settling on its own to cause clinical disease. The therapeutic dose is about 1.0 to 100 mg / kg, preferably about 1.0 to 25 mg / kg and more preferably about 2.5 to 10 mg / kg, about once a month, once every two months. Or on an intermittent schedule once every two weeks. Higher range is necessary especially when resistant (eg when an animal is infected with resistant species). Dosage levels and duration of treatment required are known to those skilled in the art. Preferred therapeutic dosages for horses with EPM or cows with neosporosis are about 1.0 to 25 mg / kg triazinetrione at 28 day intervals, with a more preferred range of about 2.5 to 10 mg / kg.

For single high dose treatment, triazineone is administered in a pharmaceutically effective amount of greater than 10 mg / kg up to about 100 mg / kg. It is a distinguishing feature of the present invention that the compounds of the present invention are nontoxic and therefore can be administered at high dose levels. The advantage of high dose administration lies in the fact that no repeated dose is required, and some triazineone compounds can cause adverse side effects when administered at very high dose levels. Particular preference is given to ponazilyl which has been found to be safe and effective at doses as high as 100 mg / kg body weight.

Without being bound to any particular theory of the invention, the unexpected success of the treatments described herein is believed to be due to the ability of the triazineone compound to cross the blood-brain barrier or placental barrier. The compounds of the present invention are believed to rescue protozoa at that location in the brain and cerebrospinal fluid / spinal cord by being able to easily cross the blood-brain barrier and also through the placental barrier. In addition, compounds of this family have been found to be nontoxic and nonmutagenic, even at the high doses required for a single high dose treatment regimen described herein.

To date, no cost-effective and easy-to-administer drugs have been marketed to effectively treat and protect these diseases without causing unacceptable side effects such as toxicity or mutagenesis in animals. The description of the triazionone compound hereafter is specifically but not limited to the tortrazilyl compound. These descriptions and claimed invention also include other triazineone compounds useful in the manner of tortrazilyl compounds. Tortrazilyl compounds useful herein are compounds of general formula (I) and physiologically acceptable salts thereof:

Where

R ¹ represents halogenoalkylthio, halogenoalkylsulfinyl or halogenoalkylsulfonyl,

R ² represents an optionally substituted sulfamoyl such as hydrogen, alkyl, alkoxy, alkoxyalkyl, alkylmercapto, halogen, halogenoalkyl, or dialkyl sulfamoyl radicals,

R ³ and R ⁴ are the same or different and represent hydrogen, alkyl, alkenyl or alkynyl,

X represents O or S.

It has also been found that especially compounds of the general formula (la) and physiologically acceptable salts thereof may be useful herein:

Where

R ^I represents halogenoalkyl (C ₁ -C ₄ ) -thio, halogenoalkyl (C ₁ -C ₄ ) -sulfinyl or halogenoalkyl (C ₁ -C ₄ ) -sulfonyl,

R ^II is hydrogen, alkyl (C ₁ -C ₄ ), alkoxy (C ₁ -C ₄ ), halogen, alkoxy (C ₁ -C ₄ ) alkyl (C ₁ -C ₄ ), alkyl (C ₁ -C ₄ ) -Mercapto, dialkyl (C ₁ -C ₄ ) aminosulfonyl or halogenoalkyl (C ₁ -C ₄ ),

R ^III and R ^IV are the same or different and represent hydrogen, alkyl (C ₁ -C ₄ ) or alkenyl (C ₂ -C ₄ ),

X represents O or S.

Finally, according to the present invention

(a) 1- (4-phenoxy-phenyl) -1,3,5-triazine of formula (I) is a compound of formula (II) wherein substituted carbonyl isocyanate of formula (III) Obtained by reacting with and optionally separating the substituted 1,3,5-triazine derivative of formula (IV) below formed during this process and then optionally reacting with a compound of formula (V)

(b) 1- (4-phenoxy-phenyl) -1,3,5-triazine of general formula (I) is a compound represented by the following general formula (II), optionally in the presence of an acid acceptor Obtained by reacting with bis- (chlorocarbonyl) -amine of

(c) A compound of formula (I) wherein R ² , R ³ and R ⁴ and X have the meanings mentioned above and R ¹ represents halogenoalkylsulfinyl or halogenoalkylsulfonyl, is represented by the following general formula (VII) It has been found that the compound of) is obtained by reacting an appropriate amount of a suitable oxidizing agent with:

Where

R ¹ , R ² , R ³ , R ⁴ and X have the meanings mentioned above,

R ⁵ represents a halogen atom, an alkoxy group or an aryloxy group,

A represents alkyl, alkenyl or alkynyl,

Z represents halogen,

R ⁶ represents alkyl,

R ^{1 ′} represents halogenoalkylthio.

When N- [3-chloro-4- (4'-trifluoromethylthio-phenoxy) -phenyl] -N'-methylurea and chlorocarbonyl isocyanate are used in process (a), the reaction procedure is as follows. It can be represented by the scheme:

N- [3-ethoxy-4- (4'-trifluoromethylthio-phenoxy) -phenyl] -thiourea and N-methyl-bis- (chlorocarbonyl) amine are starting materials in process (b). When used as, the reaction process can be represented by the following scheme:

The compound of formula (I) obtained according to process (a) or (b), wherein R ₁ is halogenoalkylthio and X is O, corresponds to the corresponding halogenoalkylsulfy according to method (c) It can be oxidized to a silyl or halogenoalkylsulfonyl derivative. When hydrogen peroxide is used as the oxidizing agent, the reaction process can be represented by the following scheme:

In general formulas (I), (II), (IV), (V), (VI) and (VII), the alkyl defined in R ² , R ³ , R ⁴ , R ⁶ or A is preferably 1 to 6 or in particular straight or branched chain alkyl having 1 to 4 carbon atoms. As examples thereof, optionally substituted methyl, ethyl, n- and i-propyl, and n-, i- and t-butyl can be mentioned.

In general formulas (I), (II), (IV), (V) and (VII), the alkenyl as defined in R ³ , R ⁴ or A is preferably 2 to 6, in particular 2 to 4 carbon sources Linear or branched alkenyl having a ruler. As examples thereof, optionally substituted ethenyl, propen-1-yl, propen-2-yl and buten-3-yl may be mentioned.

In general formulas (I), (II), (IV), (V) and (VII), the alkynyl as defined in R ³ , R ⁴ or A is preferably 2 to 6, in particular 2 to 4 carbon sources Linear or branched alkynyl having a ruler. As examples thereof, mention may be made of optionally substituted ethynyl, propyn-1-yl, propyn-2-yl and butyn-3-yl.

In general formulas (I), (II), (III), (IV) and (VII), the alkoxy as defined in R ² or R ⁵ is preferably a straight chain having 1 to 6, in particular 1 to 4 carbon atoms Or branched alkoxy. Examples thereof may be mentioned methoxy, ethoxy, n- and i-propoxy, and n- and i-butoxy.

In the general formulas (I), (II), (III), (IV), (V) and (VII), the halogen as defined in R ² , R ⁵ or Z is preferably fluorine, chlorine, bromine and iodine, Especially chlorine and bromine.

In the general formulas (I), (II), (IV) and (VII), the halogenoalkylthio defined in R ¹ is preferably 1 to 4, in particular 1 or 2 carbon atoms and preferably 1 to Halogenoalkylthio having 5, in particular 1 to 3, identical or different halogen atoms, halogen being preferably fluorine, chlorine and bromine, in particular fluorine and chlorine. Examples thereof may be mentioned trifluoromethylthio, chlorodifluoromethylthio, bromomethylthio, 2,2,2-trifluoroethylthio and pentafluoroethylthio.

In the general formulas (I), (II) and (IV), the halogenoalkylsulfinyl as defined in R ¹ is preferably 1 to 4, in particular 1 or 2 carbon atoms and preferably 1 to 5, In particular halogenoalkylsulfinyl having 1 to 3 identical or different halogen atoms, halogen being preferably fluorine, chlorine and bromine and iodine, in particular fluorine and chlorine. Examples thereof include trifluoromethylsulfuryl, chlorodifluoromethylsulfuryl, bromomethylsulfinyl, 2,2,2-trifluoroethylsulfinyl and pentafluoroethylsulfinyl.

In the general formulas (I), (II) and (IV), the halogenoalkylsulfonyl defined in R ¹ is preferably 1 to 4, in particular 1 or 2 carbon atoms and preferably 1 to 5, Especially halogenoalkylsulfonyl having 1 to 3 identical or different halogen atoms, halogen being preferably fluorine, chlorine and bromine, in particular fluorine and chlorine. Examples thereof include trifluoromethylsulfonyl, chlorodifluoromethylsulfonyl, bromomethylsulfonyl, 2,2,2-trifluoroethylsulfonyl and pentafluoroethylsulfonyl.

In formulas (I), (II) and (IV), the optionally substituted sulfamoyl as defined in R ² is preferably one of the following radicals:

In general formula (III), the aryloxy as defined in R ⁵ is preferably monocyclic carbocyclic aryloxy or acyclic carbocyclic aryloxy, in particular phenoxy.

In general formula (III), aryloxy R ⁵ is preferably phenoxy.

Most of the substituted ureas or thioureas of the general formula (II) used as starting materials are not yet known, but they correspond to (a) substituted 4-aminodiphenyl ethers in inert solvents at temperatures of 0 to 100 ° C. React with substituted isocyanates or isothiocyanates, or vice versa, (b) ammonia or substituted amines and the corresponding substituted isocyanato or 4-isothiocyanato-diphenyl ethers. Are reacted with each other under the same conditions, or (c) substituted 4-hydroxyphenyl-urea or -thiourea in an aprotic solvent such as dimethylsulfoxide, dimethylformamide or hexamethylphosphate triamide Condensation reaction with activated halogenoaromatic compounds, for example, at temperatures of 20 to 150 ° C. in the presence of sodium hydride, potassium hydroxide and potassium carbonate zam. As it can be easily produced by a method known per se.

With appropriate selection of the amount of solvent, the reaction product generally precipitates out upon cooling the solution. Another method for preparing urea from amines and isocyanates is described in Method der Org. Chemie (Methods of Organic Chemistry) (Houben-Weyl), IVth edition, Volume VIII, pages 157-158.

Some of the bis- (chlorocarbonyl) -amines of formula (VI) that can be used according to the invention in process (b) are already known (see Synthesis 1970, page 542-543) and are not yet known. In this case, they can be prepared from cyclic diacyl disulfide by chlorination in an inert organic solvent, preferably carbon tetrachloride, in a similar manner.

Reaction of urea or thiourea of formula (II) with carbonyl isocyanate (method a) of formula (III) and bis (chlorocarbonyl) -amine (method b) of formula (VI), and also of general formula ( Possible diluents for the reaction of the 1,3,5-triazine derivatives of IV) with the compounds of the general formula AZ are all organic solvents which are inert to this reaction.

These include, in addition to pyridine, preferably aromatic hydrocarbons such as benzene, toluene and xylene, halogenated aromatic hydrocarbons such as chlorobenzene and dichlorobenzene, and ethers such as tetrahydrofuran and dioxane.

Hydrochloric acid, which may form during the reaction, may be released as a gas or may be combined with an organic or inorganic acid acceptor. As the acid acceptor, pyridine which is preferably a tertiary organic base, for example trialkylamine, for example triethylamine, N-hetero mono- or non-cyclic aromatic amine, for example mono- or non-cyclic Aza-cycloalkylamines such as diazabicyclononene, diazabicycloundecene and the like or inorganic bases such as alkali metal carbonates, oxides or hydroxides, or alkaline earth metal carbonates, oxides or hydroxides .

The reaction temperature of the above-mentioned reaction step can vary within a wide range. In general, the reaction is carried out at about 0 to about 150 ° C, preferably about 20 to about 100 ° C.

In the above-mentioned reaction step, the reaction can be carried out at atmospheric pressure or under elevated pressure. In general, the reaction is carried out at atmospheric pressure.

According to method (c), the oxidizing agent capable of converting the trifluoromethylthio compound of formula (I) in which Y represents oxygen, to the corresponding sulfinyl or sulfonyl is suitably H ₂ O ₂ / glacial acetic acid; H ₂ O ₂ / acetic anhydride; H ₂ O ₂ / methanol; Peracids such as m-chloroperbenzoic acid and chromic acid; Potassium permanganate; Sodium periodate, cerise ammonium nitrate; And nitric acid.

The resulting compound can be converted to the corresponding addition salt, for example by reaction with an inorganic or organic base.

In carrying out the present invention, triazineone compounds may be formulated in a manner convenient for administration to an animal. Preferred formulations for oral administration in the present invention are preparations in the form of suspensions, tablets, capsules, gels, pastes, cyclic pills, or powders, granules or pellets. Preferred formulations for oral administration are pastes or food additives. Other routes of administration that can be used include parenteral, topical, intramuscular and intramucosal dosage forms, or other routes known in the art. Topical administration in the form of drops is also preferred.

Typically, pharmaceutically acceptable carriers and auxiliaries are used in the formulation. Examples thereof include Carbopol, an inorganic thickener, for example a thickening agent selected from the group consisting of silicates, bentonite or colloidal silica and organic thickening agents, for example fatty alcohols or fatty acid esters, and polyethylene glycol And wetting agents selected from the group consisting of sodium lauryl sulfate, Carbopol, more specifically Carbopol 974P, is the most preferred thickening agent in the preferred paste formulations herein. Also used herein may be preservatives selected from the group consisting of parabens, alcohols and aldehydes. These may be liquid, solid or gaseous substances which are inert or pharmaceutically acceptable and compatible with the active ingredient.

Surprisingly, the pastes according to the invention are effective in treating parasites. More specifically, the paste of the present invention delivers triazionone, particularly tortrazilyl and ponazilyl, through the blood-brain or placental barrier to attack parasites that have infected or invaded the fetus of an already pregnant animal. It is surprising to be effective. For convenience, certain embodiments of the pastes preferred herein are provided herein and methods for making the same. Preferred pastes according to the present invention contain triazinetrione (for example ponazilyl), propylene glycol, thickening agents such as carbopol, preservatives such as methylparaben and propylparaben and micronized suspensions of water. It can be prepared by combining water, typically purified water, with propylene glycol to heat the blend to about 70 ° C. and then adding a preservative at this temperature. After cooling the resulting mixture to room temperature, Carbopol, preferably Carbopol 974P is added. Finally, triazinetrione is added. After completion of mixing, adjust the pH to about 6.0 with sodium hydroxide. The most preferred pastes are 15% w / w ponazuryl, 20% w / w propylene glycol, Carbopol 974P 0.5% w / w, methylparaben 0.14% w / w, propylparaben 0.02% w / w, sodium hydroxide 0.1% w / w, the remainder being purified water. Sweeteners can be added to improve taste, including dextrose, sucrose, lactose, fructose, sorbitol, xylitol, artificial sweeteners and molasses. In addition, yeast or soy sauce flavoring agents may be added for the same purpose.

The invention is described in more detail in the following examples, but is not limited to these examples.

Example

Example 1 :

Pharmacokinetic investigation was performed by comparing the blood concentrations of tortrazilyl, ponazilyl and tortazuryl sulfoxide at various time points after a single dose of tortrazil was administered to the horse. All horses were orally administered a single dose of 10 mg / kg in suspension. Blood samples were taken at treatment (0) and at 0.25, 0.5, 1, 2, 4, 6, 12, 24, 48 and 72 hours after treatment. The sampling results are shown in Table 1 below. Surprisingly, the serum concentration of ponazilyl is relatively high in horses that have been treated with tortrazil. In addition, significant levels of tortrazyl sulfoxide were found in the bloodstream. This sarcoidosis when seutiseu neuro or (Sarcocystis neurona), Toxoplasma gondi (Toxoplasma gondii), neohseupo la Carney num (Neospora caninum) and neohseupo La required to treat neurological diseases such as diseases caused by the recess when (Neospora hugesi) One characteristic is that the passage of blood-brain barrier indicates that ponazilyl alone has reached satisfactory blood levels.

Table 1

Pharmacodynamics After Single Dose of Tolatrazil in Horses

Table 1-Continue

Example 2 :

Phenazuryl, 1-methyl-3- [4-p- (trifluoromethyl) sulfonylphenoxy] -m-tolyl] -s-triazine-2,4,6- (1H, which is a representative triazinetrione , 3H, 5H) -trione was formulated into a paste for horse administration. The ingredients shown in Table 2 were used in preparing the formulations as follows.

TABLE 2

Ingredients of Phonazil Horse Paste

ingredient Theoretical quantity Substantial amount (% w / w) Micronized Ponazylpropylene Glycol Carbopol 974P Methylparaben, NF Methylparaben, NF Sodium Hydroxide, NF 22.5 kg 30.0 kg 0.750 kg 0.210 kg 0.030 kg 0.150 kg 96.365 kg 15.020.00.50.140.020.1064.24

Formulations were prepared using methods (A) and (B) as follows:

The first method (A) is as follows: 1) mixing some water with propylene glycol; 2) after addition of preservatives (methylparaben and propylparaben); 3) Slowly add Carbopol 974p until a uniform suspension is obtained; 4) ponazilyl is added in micronized form; 5) add sodium hydroxide so that the pH of the suspension is about 6.0; 6) The remaining water was added to a sufficient volume. The final suspension is in the form of a paste that can be delivered orally to horses.

The second method (B) is as follows: 1) a portion of water is mixed with propylene glycol; 2) after heating to 70 ° C .; 3) preservatives (methylparaben and propylparaben) are added while maintaining the solution at 70 ° C; 4) cooling the solution to room temperature; 5) Slowly add Carbopol 974p until a uniform suspension is obtained; 6) ponazilyl is added in micronized form; 7) add sodium hydroxide so that the pH of the suspension is about 6.0; 8) The remaining water was added to a sufficient volume. The final suspension is in the form of a paste that can be delivered orally to horses.

The suspension obtained was administered to horses and ate well.

Example 3 :

Phenazuryl, 1-methyl-3- [4-p- (trifluoromethyl) sulfonylphenoxy] -m-tolyl] -s-triazine-2,4,6- (1H, which is a representative triazinetrione , 3H, 5H) -trione was tested for the ability to treat horses already proven to have protozoan encephalomyelitis (EPM) symptoms in horses. The compound was formulated into a paste using ponazilyl as the 15% active ingredient described in Example 1. It was administered once a day for 28 days to horses already found to be EPM at a dosage rate of 2.5 to 10 mg / kg.

Naturally occurring clinical cases of EPM have already been characterized by impression statements and laboratory diagnostics. The diagnostics used to introduce EPM-positive horses into the experiments were as follows: certain asymmetric neurological defects as determined by standardized neurological examination, including: radiographs showing EPM; Positive Western blot for Sarcocytis neurona lgG; Erythrocyte cell numbers up to 500 cells / ml; CSF index-total protein <90, lgG index> 0.3, AQ ratio <2.2.

An additional condition is that the horse does not suffer from diseases other than EPM. Therefore, they must meet the following criteria: negative CSF for EHV-1 (<1: 4); Normal serum levels of vitamin E (.2.0 μg / ml); No stroke disease; No behavioral disorders.

The diagnosed horses were divided randomly into groups. The first group horses were administered a paste formulation at a dose of 5 mg / kg per day; The horses of the second group were administered a paste formulation at a dose of 10 mg / kg per day. The amount of treatment was based on body weight. 90 days (approximately 60 days after discontinuation of treatment) were evaluated to see if the treatment for horses was effective. Response to treatment was scored according to the following system:

1) 0 = completely successful—CSF is clinically normal as negative; 2) 1 = slight defect observed in normal gait; 3) 2 = excessive gait, turning, shaking, jaw lumbar pressure and neck dilatation and defects easily observed; 4) 3 = Defects with very excessive gait, facial swing, lumbar pressure or neck dilatation; 5) 4 = unconsciously stumbles, walks faster, falls; 6) 5 = can't lie down. Improvement of one (1) unit in the score is considered a significant improvement.

The results of the experiment are shown in Table 3 below. Horses in the 10 mg / kg group treated for 28 days showed significant improvement in clinical scores from day 0 to day 90 after treatment with all (100%) ponazilyl. Eight out of nine (88.9%) of horses treated at a dose of 5 mg / kg showed a satisfactory improvement. For each treatment day, the sum of each group's scores results in a total score. The improvement in the total score demonstrated by both the first group and the second group is nearly equivalent. Thus, it can be concluded that administering ponazilil at 5 mg / kg or 10 mg / kg is effective for actively treating EPM in horses.

Table 3 : Responses of EPM-Infected Horses When Treated With Tolatrazil Sulfone

Example 4 :

In vitro tests were performed to determine the extent of protection provided by ponazil. The following parasite strains were evaluated for susceptibility to the ponazilyl compound: SN3 strain of Sarcocytis neuron; SF1 strain of Sarcocystis falcatula ; RH strain of Toxoplasma gondi; And NC-1 strain of neospora caninum. Ponazuryl was tested at two concentrations (1 μg / ml and 10 μg / ml).

Bovine turbinate (BT) cells were used for all in vitro tests. Cells were RMPI supplemented with 10% v / v fetal bovine serum (FBS), 100 units penicillin (g / ml), streptomycin 100 mg / ml and 5 × 10 ⁻² mM 2-mercaptoethanol in a 25 cm 2 flask Proliferation to fuse in 1640 medium. After obtaining the cell fusions, cells were maintained in the same medium except with reduced FBS (2% v / v). Cell cultures were incubated at 37 ° C. in a humid atmosphere containing 5% carbon dioxide and 95% air.

To grow parasites, BT cell monolayers were infected with parasites and examined by microscopy for the development of lesions (cytopathic effect, "CPE") or the presence of a number of extracellular mesozoites. If lesions were observed or there were a number of extracellular parasites, the monolayer was scraped off with the tip of a 5 ml pipette and one to three drops of fluid containing mesozoite were introduced into two flasks with new BT cells. The mesozytes of Sarcocistis Neurona and Sarcocistis Falcatula were passed in the same manner every 5 to 10 days, and the tachyzoites of Toxoplasma gondic and Neospora caninum were passed every 3 to 4 days.

The assay used to determine the effectiveness of ponazil is microtiter tomographic failure analysis (MMDA). This assay was used to determine whether the parasite or compound is toxic to BT cells. Inoculate BT cells into flat-bottomed 96-well microtiter plates and use the resulting monolayer to determine the effect of tortrazil and ponazilyl on merozoid production as measured by CPE (plaque formation). saw. Monolayers were inoculated with parasites (Sarcocystis neurona or Sarcocistis Falcatula at 50,000 / well, Toxoplasma gondic at 10,000 / well and Neospora caninum at 20,000 / well). After 2 hours of inoculation, all wells were inoculated with test compounds. Untreated and uninfected monolayer wells served as parasite controls and BT cells treated with uninfected agents served as toxic controls. Each treatment was tested at 6 replicates. Each well was visually observed daily and the test was stopped when 90-100% of untreated mesozoite infected cells were lysed (90-100% CPE). All wells of the plates were rinsed with phosphate buffered saline (PBS), fixed in 100% methanol for 5 minutes and then stained with crystal violet solution. Areas of destruction by BT cell death or mesozoites due to toxicity do not absorb crystal violet. The ELISA plate reader was used to quantify the amount of crystal violet absorption and these data were used to determine the concentration of ponazilyl (inhibition concentration ₅₀ or IC ₅₀ ) that inhibits destruction by 50%. Data demonstrating inhibition is shown in Table 4 below. From this, 100% inhibition of cell destruction by Ponazilillo Neospora caninum, Toxoplasma Gondi, and Sarcocytis Falcatula, which is small in amounts of about 1 μg / ml, whereas cell destruction by Sarcocytis neurona was 100%. It can be seen that 10 μg / ml of ponazilyl is required to inhibit%. This is accompanied by neurological and dysplastic diseases, including triazines such as tortrazilyl and ponazilyl, caused by Sarcocytis neurona, neospora caninum, neospora resting and toxoplasma gondic. It is shown to be effective in treating a disease caused by known coccidia. In addition, ponazilyl is nontoxic to BT cells.

Table 4 :

In vitro data for Ponazil

Example 5 :

This experiment was conducted to see if triazine, such as tortrazil, could cross the blood-brain barrier. Normal horses were divided into three groups of three horses per group. To horses of the first group were orally administered tolstrazilol at a dose of 2.5 mg / kg as a 5% suspension. Group 2 horses were orally administered tolstrazilol at a dose of 5.0 mg / kg as a 5% suspension. The third group horses were orally administered tolstrazilol at a dose of 7.5 mg / kg as a 5% suspension. Dosing was repeated daily for 10 days. Blood samples were taken at 48, 96 and 240 hours to determine the concentrations of tortrazilyl, tortrazil sulfoxide and ponazilyl in serum. Ten days after initiation of treatment (day 10), cerebrospinal fluid samples were taken from each horse and the concentrations of tortrazil, tortazuryl sulfoxide and ponazilyl were remeasured in these samples. The concentrations of tortrazil, tortrazil sulfoxide and ponazuryl in serum and cerebrospinal fluid are shown in Tables 5a and 5b. After treating horses with tortrazil, the concentration of ponazilyl in the cerebrospinal fluid is essentially the same as that of tortrazuril itself. The concentration is significant. This demonstrates that both tortazuril and ponazuril cross the blood-brain barrier more effectively, and that ponazuril crosses the blood-brain barrier more effectively than toltrazuril. This data also suggests that triazineone can effectively cross the placental barrier to those skilled in the art.

Table 5a : Drug Levels After Repeated Administration of Toltrazolil to Horses

Table 5b : Drug levels after repeated doses of tortrazil to horses

Although the invention has been described in detail above for purposes of illustration, it is for the purpose of illustration only and is not to be departing from the spirit and scope of the invention by those skilled in the art, except as may be limited by the claims. It can be modified in

Claims (25)

A pharmaceutically effective amount of a triazineone compound is administered to an animal suffering from parasitic neurologic or abortigenic disease exhibiting sensitivity when treated with the triazineone compound. A method of therapeutically treating a suffering animal, except when the disease is Sarcocystis neurona , except when the compound is diclazuil or toltrazuril . The method of claim 1, wherein the parasitic disease is caused by Coccidia . The method of claim 2, wherein coccidia is a member of the Sarcocystidae family. The method of claim 3, wherein when a member of the family in sarcoidosis sarcoidosis cis peptidase seutiseu (Sarcocystis), neohseupo la is selected from the group consisting of (Neospora), and toxoplasmosis (Toxoplasma). The method according to claim 4, wherein sarcocytis is selected from the group consisting of Sarcocystis spp. , Neospora is selected from the group consisting of Neospora spp. , And toxoplasma is toxoplasma species. ( Toxoplasma spp. ). 6. The method of claim 5, sarcoidosis when seutiseu paper sarcoidosis when seutiseu a neuro or (Sarcocystis neurona), and neohseupo La paper neohseupo la Carney num (Neospora caninum) or neohseupo la rest upon (Neospora hugesi), Toxoplasma paper Toxoplasma gondi ( Toxoplasma gondii ). 5. The method of claim 4, wherein sarcocistis is a sarcocistis neurona that induces Equine Protozoal Myeloncephylitis. The method of claim 4, wherein the neospora is neospora caninum that causes bovine or canine Neosporosis . The method of claim 4, wherein the toxoplasma is Toxoplasma gondii . The disease is characterized by administering a metaphylactically effective dose of a triazineone compound to an animal infected with a parasite, which is a cause of neurological or dysplasia that exhibits sensitivity when treated with a triazineone compound. Meta-protective treatment of animals infected with parasites, the causative agent of. The method of claim 1 or 10, wherein the triazionone compound is selected from the group consisting of tortrazilyl, ponazilyl, and diclazuryl. The method of claim 1 or 10, wherein the triazionone compound is ponazilyl. The method of claim 1 or 10, wherein the triazineone compound is administered in two or more repeat doses. The method of claim 13, wherein the repeat dose is administered in an amount from 1.0 to 100 mg / kg. The method of claim 10, wherein the triazineone compound is administered until protective immunity develops in the animal. The method of claim 1 or 10, wherein the triazineone is administered in an amount of 2.5 to 10 mg / kg. The method of claim 16, wherein the triazineone is tortrazilyl or ponazilyl. The method of claim 1, wherein the triazineone compound is administered in a single high dose of greater than 10 mg / kg. The method of claim 8, wherein the triazineone compound is administered in repeated regular doses until immunological protection is established. The method of claim 1, wherein the triazineone compound is administered at a dosage of 2.5 to 10 mg / kg daily for a 28 day period. A therapeutic composition comprising (a) a pharmaceutically effective amount of a triazineone compound, (b) a carrier, and optionally (c) an adjuvant for treating an animal having a disease susceptible to the triazineone compound. The composition of claim 21 in the form of a paste. A method of treating EPM, characterized by administering a therapeutically effective amount of one or more triazinedione to a horse that is expected to suffer from horse protozoan encephalomyelitis (EPM). The method of claim 23, wherein the triazinedione is diclazilyl. The method of claim 23 wherein the horse is horses.