JPH0320376B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to compounds and preparations and methods for making these compounds for the treatment and prevention of theileriosis in cattle and sheep. Theileriasis is a disease caused by protozoa of the genus Theileria . In cattle, T. Parva (parva),
T. Parval (lawrencei), T. Parva (Bovis), T.
Sergenti, T. annulata and T. mutans are responsible for substantial losses mainly in Central and East Africa and the Middle East. In sheep, T. hirci and
Ubiquitous in the Middle East caused by T. ouis. Infected ticks transmit these protozoa to mammalian hosts. When Hinoba bulb cells are infected, they immediately begin to divide, contrary to their normal function. Mortality occurs due to the release of toxic substances from the destroying bulbs and the harmful effects of the parasite itself. After being released by the bulb, the Theileria parasites infect red blood cells and ticks, which in turn infect animals. In the field, there is no known effective treatment for theileriosis. Compounds with anti-Theilerial activity are known but are toxic to the host. They are not used to treat Theileria infections. The only drug ever used to treat cattle and sheep is oxytetracycline (Terramycin). It is commonly used for Anaplasma infections and bacterial infections. 1968Academic
Press, edited by Weinman D. and Ristic M., SF
Infectious Blood Diseases of Man by Barnett
and Animals, Vol., this compound is
They have very limited effectiveness when administered before infection has established, but are ineffective once infection has established. Often, Anaplasma infections are misdiagnosed and diagnosed as Theileriosis, resulting in the administration of oxytetracycline, often in large doses to almost toxic levels. What appears to be an apparent recovery from theileriosis is actually just a recovery from anaplasmosis and bacterial infection. British Patent No. 1553424 describes 2-hydroxy-3-cyclohexylalkyl and 2-hydroxy-3-cyclohexyl-1,4-naphthoquinone derivatives useful in the prevention and treatment of theileriosis. It was subsequently found that the latter, 2-hydroxy-3-cyclohexyl-1,4-naphthoquinone, is a very effective compound, especially against infections with T. parva . European Patent Application No. 78101426 may be
Referring to 3-hydroxy-1,4-naphthoquinones whose 2-position substituent is a C _3-12 cycloalkyl ring bearing a substituent such as a C _1-4 alkyl group, especially a methyl group. There is. However, a specific compound having a substituent is not described, nor is it described at which position on the cycloalkyl ring the cycloalkyl ring should be substituted. The compound of formula () below is our patent application.
It was described in UK No. 8131206 as an intermediate in the synthesis of certain 2-substituted-3-hydroxy-1,4-naphthoquinones. Now, the compound below () is
It shows high and favorable activity against various Theileria species in vivo and in hydro conditions and, in contrast to the compound of UK Patent No. 1553424, is particularly effective against T. annulata infections. It's been found. As a result, it becomes possible to produce new therapeutic compounds and to treat infected animals against diseases that occur in these organisms. As one feature of the present invention, the formula (wherein R is an alkyl group having 1 to 10 carbon atoms), tautomers thereof, or pharmaceutically acceptable salts thereof. As a result of tautomerism, the 3-hydroxy group donates its acidic hydrogen to the adjacent oxo group, but the formula ()
is believed to represent a more stable state. Since the hydroxy group in the formula is capable of forming salts with suitable bases, pharmaceutically acceptable salts of the compounds include salts of alkali metals such as sodium or potassium and organic bases such as ethanolamine, diethanolamine and N-methylglucamine. There is salt. As can also be seen, the compound of formula () is
It can exist as cis or trans isomers and the cyclohexyl ring can be cis or trans substituted with naphthoquinone-methylene residues and 4-t-alkyl groups. The present invention is directed to both cis and trans isomers and mixtures thereof in any proportion. Note that in the unsubstituted 1,4-naphthoquinone ring the 2 and 3 positions are the same, so a cyclohexyl substituent or a hydroxy group can be placed at the 2 position. For convenience herein, when referring to compounds generally, the substituent cyclohexyl substituent will be at the 2-position. In the compound of formula (), it is appropriate that R is a linear C _1-4 alkyl group. Preferred groups are methyl or ethyl. Compounds of formula () may be prepared by known methods for the preparation of compounds with similar structures. For example, a compound of formula () may be prepared from a compound of formula () where R is as defined by standard literature methods, converting it into a 3-hydroxy-1,4-naphthoquinone. Compounds of formula () may be prepared from compounds of formula (A) by Fries rearrangement. Alternatively, compounds of formula () can be prepared by adding the corresponding 3-halogeno, i.e., 3-chloro- or 3-brome, analogs of formula (), where X is halogen and R is as defined above, to an alkali. Hydrolysis, for example with alkali metal hydroxides in a suitable medium, can convert the 3-hydroxy substituted compounds. For example, it has been found convenient to use potassium hydroxide in an aqueous methanol medium. The starting 3-halogeno derivative is the method suggested by Fierser, L. ( J. Am.) starting from substituted or unsubstituted 2-cyclohexylmethylphenol.
Chem. Soc., 1948, 3165), correspondingly substituted 4a,5,7,7a-tetrahydro-2
Obtained from -cyclohexylmethyl-1,4-naphthoquinone by oxidation and halogenation. Expression() can be used as a further alternative, expression() (However, R is as defined above.) It can be produced by introducing a hydroxyl group into the 3-position of the compound. Thus, for example, a compound of formula () can be halogenated (ie, chlorinated or brominated) to give a compound of formula () above, which in turn can be made into a compound of formula () above. The compound of formula () is epoxidized to form formula () This can be hydrolyzed, for example, with a dilute base or dilute aqueous acid solution to give a compound of formula (). Compounds of formula () may also be subjected to Thiele acetylation to give compounds of formula (a) by reaction with an acylating agent (eg acetic anhydride) in the presence of an oxidizing agent (eg perchloric acid). Compounds of formula (a) are hydrolyzed to give compounds of formula (b), which can be oxidized to compounds of formula () in a manner similar to that described, for example, in Organic Reaction, Vol. 19, p. 222. Suitable oxidizing agents include, for example, ferric chloride
Iron-mineral acids or chromic acids. In the method described above, the stereochemistry of the final product is determined by that of the starting materials. That is, using a stereochemically pure compound in a reaction will yield a pure cis or trans isomer. As a more advantageous alternative, a compound of formula () (wherein R ¹ is a halogen, for example, a chlorine or bromine atom, or a hydroxy group, an acetoxy group or an alkoxy group) and a compound as defined above in a form capable of providing a substituted cyclohexylmethyl group as a free radical. react with a cyclohexylmethyl group-donating compound containing an R bond and, if desired, R ¹ other than a hydroxy group in the resulting 2-cyclohexylmethyl group-substituted condensate.
It can also be produced by hydrolyzing the group to form a 3-hydroxyl group. Preferred donor compounds are correspondingly substituted cyclohexyl acetic acids which are capable of undergoing oxidative decarboxylation. For example, persulfuric acid with catalysts such as silver ions is used for this purpose (Jacobson N. et al.
Annalen, 1972, 763, 135 and Acta Chem.
Scand.1973, 27, 3211). Preferably, when using persulfuric acid under these conditions, 1,4-naphthoquinone in which R ¹ is substituted with a group other than a hydroxy group is used. Conveniently, ammonium persulfate can be used as the oxidizing agent and the catalyst is silver nitrate. The main coupling reaction is followed by hydrolysis, if necessary, to yield hydroxyl groups. Alkaline conditions are usually advantageous for hydrolysis. An example of a cyclohexylmethyl donor compound that itself has a peroxide group is suitably substituted cyclohexylmethyl acanoyl peroxide, as suggested in US Pat. No. 2,553,647. Donation of the cyclohexyl free radical by spontaneous release from the donor compound can be accomplished using, for example, tricyclohexylmethyl-borane. Such reagents may be prepared by reacting cycloalkenes with borane dimethyl sulfide. It is convenient to carry out the reaction in a solvent such as tetrahydrofuran. The 2 required for some of the above reaction schemes
-Substituted-1,4-naphthoquinones can be prepared by known synthetic methods described in the literature. For example, Fieser,
L., J., Am. Chem. Soc., 1948, 3165 or similar methods. Furthermore, the corresponding substituted cyclohexenylcarboxylic acids can also be used as cyclohexyl-donating compounds. In that case, as will be clear to the expert, an additional step of reduction of the alkene is required after the condensation to provide the compound of formula (). It is natural that the above process gives a mixture of cis and trans isomers. When a single compound of the cis or trans isomer of formula () is required, the isomerically pure starting compound is used in the above reaction in which no isomerization takes place. In particular, stereospecific pure carboxylic acids are used in the last method above. If a mixture of isomers occurs, the mixture may be separated by physical means. Such methods are known in the art, such as fractional crystallization and chromatographic separation. Yet another feature of the invention is to provide compounds of formula (). (However, in the formula, R is as defined above,
R ₁ is as defined above or is a benzyloxy group) These compounds are useful as intermediates in the above synthesis. Compounds having as R ¹ a group that is easily hydrolyzed to a hydroxyl group in vivo are:
It may be used as a prodrug in a formulation or as a therapeutic agent upon in situ hydrolysis to give the final product. Acetyl or benzoyl groups underwent such hydrolysis and could therefore be used as long-acting precursors of active compounds. The compounds of formula () have extreme activity against protozoa of the genus Theileria and can be used for the treatment and/or prevention of Theileriasis in cattle and sheep. especially,
These compounds are effective against T. annulata infection. The amount of a compound of formula () required for use in the treatment or prophylaxis of a Theileria infection will vary with the nature of the infection and the route of administration, as well as the type of active compound. As used herein, "effective amount, dose, or unit dose" refers to a predetermined dose of a compound that is sufficiently effective against Theileria microorganisms when administered to cattle or sheep in vivo. To serve or achieve prophylaxis or treatment, certain preparations may contain several times the dosage required by one animal. Initial dose for cows weighing 400Kg is 0.2 to 10
g, conveniently from 0.2 to 2.5 g, and for calves or sheep from 50 mg to 1.0 g or preferably from about 0.1
The amount of active compound is 0.5 grams, but larger doses may be used. While it is possible for the compounds of formula () to be administered as raw materials for anti-Theileriosis agents, it is advantageous to use the active ingredient as a therapeutic preparation. A therapeutic preparation containing a compound of formula ()
The composition may take the form of a sealed container or a molded tablet with ingredients such as excipients. Advantageously, the preparation is in unit dosage form or a sterile, encapsulated formulation. Preparations containing compounds of formula () may be administered by injection (subcutaneously or preferably intramuscularly), intravenously or orally in conjunction with a suitable carrier. Sterile injectable formulations may be formulated with preferably organic carriers. The carrier may contain bacteriostatic agents, antioxidants, buffering solutes to render the preparation isotonic with blood, thickening agents, suspending agents, or other pharmaceutically acceptable additives. Formulations for injection may be placed in unit-dose containers, such as ampoules, single-use injection devices, or in multi-dose forms, such as bottles, from which appropriate quantities may be dispensed. Formulations for oral administration may contain as carrier solids for the formation of tablets, pastes, granules or powders, or may contain liquids for suspension or dissolution. They include diluents, binders, dispersants, surfactants, lubricants, coating materials,
There may be colorants, solvents, thickening agents, suspending agents or other pharmaceutically acceptable additives. These preparations may be in unit-dose or multi-dose forms or as additives to foods. Compounds of formula () also serve as salt-licks, allowing animals to receive prophylactic treatment in the field. formula()
The compounds may also be made into drench formulations containing up to 10% by weight of active compound in a suitable carrier such as dimethyl sulfoxide. As yet another feature of the invention, a packaged composition for protecting and treating a mammal against theileriosis is provided. This includes preparations as defined above, together with indications for their use for protecting and treating diseases. For therapeutic purposes, the compound of formula () or a salt thereof may be given in relatively large amounts, sometimes when the animal's body temperature increases and schizoont appears, or at the time of late pathology. For the next few days, for example 5 days, the administration is repeated in the same manner every day. The total dose over this treatment period is preferably 0.5 to 50 mg/Kg,
More preferably the dosage will be from 0.5 to 10 mg/Kg and even more preferably from 1 to 5 mg/Kg. Another treatment is to administer once or twice daily for a total of 10
It can be administered multiple times. For prophylactic treatment, for example when it is suspected that an animal has been infected, administer a compound of formula () from 1 to 5 mg/Kg on the first day, then repeat the same dose once weekly, up to It may take up to 5 doses. Depending on the risk of infection, 0.2 to 5 mg/Kg may be administered per week. The period of prophylaxis can range from 4 to 20 or 120 days. Alternatively, the compound
In addition to slow-release implants, pellets can be injected under the skin with a relatively insoluble carrier. As the pellets dissolve,
The active ingredient is released slowly, for example over a period of four months, and a low level of protection is maintained, such as with the above-mentioned dosages. In yet another aspect, a method of prophylactically or therapeutically treating bovine or ovine theileriosis is provided. In this method, an effective amount of a compound of formula () or a pharmaceutically acceptable salt thereof is prophylactically or therapeutically administered to an animal infected with or likely to be infected with a pathogenic Theileria species. do. In particular, the method of treatment is accomplished using preparations or compositions containing compounds as defined above. Furthermore, another feature of the treatment as defined above is the administration of a vaccine containing one or more live Theileria species or strains that act as effective antigens. To produce vaccines for this purpose, Theileria species, for example, T.parva
(parva) and T. parva (lawrencei) (Radley,
DE et al. (1975), Veterinary Parasitology, 51−
(60) prepared from ground mites or other infected material (e.g., suspensions or cultured mite cells of Heileria- infected mammalian cells or salivary tissue infected with Theileria or their products or exudates) Conveniently, the dosage of said active compound is between 0.5 and 5 mg/Kg of the host animal;
From the date of vaccination, 7 days after vaccination or
Administer until 14 days later. The first treatment thus consists of administration of a combination product which releases the antigen immediately, followed by administration of the active compound for 7 days or more. The initial dose may be followed by weekly doses of 0.5 to 5 mg/Kg for 10 weeks. Example A Preparation of 4-t-alkylcyclohexanecarboxylic acid Carboxylic acids used in later examples were prepared as follows. 4-t-butylcyclohexane-1-carboxylic acid, 4-t-butylcyclohexyl acetic acid, and their pure cis and trans isomers are:
It is available as a commercial product or in the literature (J. Amer.
Chem.Soc.1970, 92 , 2800 and the literature described therein). 4-t-pentylcyclohexyl-1-carboxylic acid (cis/trans mixture) and pure trans-4-t-pentylcyclohexyl-1-
Carboxylic acid was produced as follows. a Production of cis/trans-4-t-pentylcyclohexyl-1-carboxylic acid 4-t-pentylcyclohexanone (49.2g)
was dissolved in ether (200ml) and sodium cyanide (24.46g) and water (30ml) were added. The mixture was cooled to 0° C. and concentrated hydrochloric acid was added over 1 hour while stirring vigorously. The mixture was stirred for an additional 3 hours and left overnight. The mixture was washed with saturated sodium metabisulfite solution (2 x 200ml) and the ether layer was dried over sodium sulfate. The solvent was distilled off under reduced pressure to obtain 54.0 g of cyanohydrin as a pale yellow oil. Cyanohydrin (54.0g) was dissolved in anhydrous pyridine (70g).
ml) and anhydrous benzene (70 ml) and heated to 0°C.
Cool and stir. A solution containing phosphoryl chloride (90ml) in pyridine (83ml) was added over 45 minutes. During this time, the temperature was maintained at 0°C. The reaction mixture was heated to reflux for an additional 30 minutes. The mixture was allowed to cool, poured into ice, stirred for 30 minutes, extracted with ether, washed with water, dried with sodium sulfate, and evaporated to dryness under reduced pressure to yield 4-t-pentylcyclohex-1-ene-1-nitrile. . 48.4g oily. 1-4
-t-pentylcyclohex-1-ene-1-nitrile (48.0g) was added to potassium hydroxide (23.3g).
was added to a mixture of a solution in water (34ml) and ethanol (150ml). The mixture was heated to reflux for 72 hours, cooled on ice, diluted with water (175ml) and acidified with concentrated hydrochloric acid. A colorless solid precipitate was collected, washed with water, and dried. The solid was partitioned between ethyl acetate and sodium hydroxide (2N), and the base layer was separated and acidified with concentrated hydrochloric acid. The resulting colorless solid was collected, washed with water, and dried to yield 33.65 g of 4-t-pentylcyclohex-1-ene-1-carboxylic acid, melting point 123-125°C. 4-t-pentylcyclohex-1-ene-1
-Carboxylic acid (33.5g) in ethanol (275ml)
and 10% palladium on charcoal (1.0 g) was added. The mixture was hydrogenated at 10 atm to absorb the theoretical amount of hydrogen. The catalyst was removed and the colorless liquid was evaporated to dryness under reduced pressure to give 4-t-pentylcyclohexyl-
1-carboxylic acid was obtained. 27.0g (cis/trans mixture). b Production of pure trans-4-t-pentylcyclohexyl-1-carboxylic acid Cis/trans mixed acid (12 g, produced by method a)
was heated in a steam bath for 16 hours in the presence of concentrated flow (60 ml). The reaction mixture was cooled and poured into ice, producing a black solid. Remove the solids and add light petroleum (40
-60°C) to dissolve most of the solids. The petroleum extract was treated with charcoal powder and evaporated to dryness under reduced pressure to give trans-1-4-t-pentylcyclohexyl-1-
Carboxylic acid was obtained. 5.7g. Melting point 92-100℃.
It is a trans isomer with a purity of 95-97% based on the NMR spectrum. Example 1 Production of 2-[trans-(4-t-butylcyclohexyl)methyl]-3-hydroxy-1,4-naphthoquinone A Production of 1-trans-(4-t-butylcyclohexyl)-acetic acid 4-t- Butylcyclohexane-1-carboxylic acid (50g, mixture of cis and trans isomers)
, potassium hydroxide (150 g) and ethylene glycol (500 ml) were heated under reflux overnight. The reaction mixture was poured onto crushed ice (2000 g) and acidified with concentrated hydrochloric acid to remove the resulting colored precipitate. The wet solid was dissolved in ethyl acetate, the aqueous layer was removed, and the organic layer was dried over magnesium sulfate. The solvent was removed under reduced pressure to give a white solid. When this is recondensed from toluene, 1-trans-(4-t-butylcyclohexyl) is produced.
-carboxylic acid (30.9 g, melting point 172-174°C). This 1-trans-(4-t-butylcyclohexyl)-carboxylic acid (35 g) was dissolved in anhydrous ether (250 ml), and this solution was mixed with lithium aluminum hydride (7.3 g) in anhydrous ether (200 ml).
The mixture was slowly added to the solution containing the solution to maintain a gentle reflux. After the addition was complete, the mixture was heated to reflux for an additional 1.5 hours. The reaction mixture was then cooled in an ice bath and ice-cold 2N hydrochloric acid (200 ml) was added. Separate the aqueous layer and wash with ether (100ml).
The combined ether extracts were washed with dilute sodium hydroxide, water, dried over magnesium sulfate, and the solvent was removed under reduced pressure. Gave 28g of colorless oil. The above 1-trans-(4-t-butyl-cyclohexyl)-methanol (28 g) was dissolved in dry acetonitrile (330 ml), anhydrous lithium bromide (28.7 g) and chlorotrimethylsilane (44.5
g) was added and the mixture was heated to reflux for 60 hours. The reaction mixture was allowed to cool, and the solvent was distilled off under reduced pressure. The residue was dissolved in ether and washed with water (100 ml), 10% sodium bicarbonate (100 ml), water (100 ml),
It was dried with MgSO ₄ and the solvent was removed under reduced pressure. The crude product was subjected to column chromatography to obtain oily 1-trans-(4-t-butylcyclohexyl)-methane bromide (15.5 g). The above bromide (15 g) was dissolved in sodium cyanide (4.83 g) in hexamethylphosphoric triamide.
In addition, the mixture was stirred overnight at room temperature. The crude reaction mixture was diluted with water (250 ml) and extracted with ether (2 x 200 ml).
did. The organic layers were combined, washed with water (3 x 100 ml), dried (MgSO ₄ ), and the solvent was removed under reduced pressure to give 1-trans-(4-t-butylcyclohexyl)methanenitrile (10.5 g) as a colorless liquid. I got it. 1-trans-(4-t-butylcyclohexyl)-methanenitrile (10g), potassium hydroxide (20g), ethanol (200ml) and water (50ml)
was heated to reflux for 65 hours. Most of the solvent was distilled off under reduced pressure, and the residue was dissolved in water (300ml) and extracted with ethyl acetate (100ml). The aqueous layer was made acidic with hydrochloric acid,
Extracted with ethyl acetate (2 x 100ml). The organic layers were combined, washed with water (200ml) and dried over _MgSO4 . The solvent was removed under reduced pressure and the solid residue was triturated with cold water to give 1-trans-(4-t-butylcyclohexyl)-acetic acid (9.5 g, melting point 93-95°C). B Production of 2-[trans-(4-t-butylcyclohexyl)methyl]-3-hydroxy-1,4-naphthoquinone 2-chloro-1,4-naphthoquinone (960 mg),
1-trans-(4-t-butylcyclohexyl)
Acetic acid (990 mg, J.Amer.Chem. as above)
Soc. 1970, 92, 2800) and silver nitrate (250 mg) in acetonitrile (9 ml) were vigorously stirred and kept under reflux, and a solution of ammonium persulfate (3.0 g) in water (12 ml) was mixed for 1 hour. It was added dropwise. The mixture is further 1
The mixture was refluxed for an hour, cooled on ice, and the resulting yellow solid was collected and washed with water. The solid was extracted with hot ethyl acetate and cooled to yield 2-[(trans-(4-t-butylcyclohexyl)-methyl]-3-chloro-1,4-
Naphthoquinone was given. Melting point 154-156℃. NMR
The spectrum is 2-[trans-(4-t-butylcyclohexyl)methyl]-3-chloro-1,4
-Shows the structure of naphthoquinone. It was also shown that it is 100% trans isomer. Melting point is 154−156
℃. A solution containing chloroquinone (6 g) obtained above in dimethoxyethane (60 ml) and water (60 ml) was heated to reflux, and a solution containing potassium hydroxide (6.0 g) in water (60 ml) was heated to reflux. Added dropwise over 10 minutes. After refluxing for an additional 15 minutes, the mixture was rapidly cooled to room temperature and acidified with concentrated hydrochloric acid. The resulting light yellow solid was collected, washed with water, and dried to yield 2-[trans-(4-t-butylcyclohexyl)methyl]-3-hydroxy-1,4-naphthoquinone. Melting point 180-182℃. Example 2 2-[cis-(4-t-butylcyclohexyl)
Preparation of 2-chloro-1,4-methyl]-3-hydroxy-1,4-naphthoquinone in a similar manner to that described in Example 1.
- Naphthoquinone is treated with 1-cis-(4-t-butylcyclohexyl)-acetic acid, the resulting chloroquinone is hydrolyzed, and 2-[cis-4-t-butylcyclohexyl)methyl]-3-hydroxy-1,4
- Naphthoquinone was obtained. Melting point 124-125℃. Example 3 2-[trans-(4'-t-pentylcyclohexyl)methyl]-3-hydroxy-1,4-naphthoquinone A Production of 1-trans(4-t-pentylcyclohexyl)acetic acid trans-t-pentylcyclohexane- 1-
Carboxylic acid (8.0g), dichloromethane (34.7ml)
and thionyl chloride (17.77 g) were stirred at room temperature for 22 hours. The reaction mixture was evaporated under reduced pressure to a black residue. This was distilled under reduced pressure to obtain trans-t-pentylcyclohexane-1-carboxylic acid chloride. 7.05g. Boiling point 107−110°/0.5
mmHg. The acid chloride was added dropwise to a solution containing diazomethane in ether over 30 minutes at 0°C.
The solution was left in the refrigerator overnight. Excess solvent was distilled off under reduced pressure to obtain the corresponding diazoketone. yellow oil
6.8g. Diazoketone (6.8g) was dissolved in anhydrous methanol (60g).
A solution containing silver benzoate (0.45 g) in triethylamine (7 ml) was added with stirring. The solution was heated under reflux for 1 hour, allowed to cool, the charcoal powder was removed, and the solution was evaporated to dryness under reduced pressure. The residue was distilled to obtain trans-t-pentylcyclohexylmethyl acetate. Boiling point 94-100℃/0.2mmH
g. Trans-t-pentylcyclohexyl methyl acetate (3.6 g) was dissolved in methanol (11 ml), and sodium hydroxide (1.08 g) was dissolved in water (5 g).
ml) was added and the reaction mixture was stirred overnight at room temperature. The reaction mixture was diluted with water (50ml) and extracted with ether. The aqueous phase was acidified with concentrated hydrochloric acid and extracted with ether. The ether extract was dried over magnesium sulfate and evaporated under reduced pressure to give oily 1-trans-(4-t-pentylcyclohexyl)acetic acid (2.0 g). B Production of 2-[trans(4't-pentylcyclohexyl)methyl]-3-hydroxy-1,4-naphthoquinone 2-chloro-1,4-naphthoquinone (0.95 g),
A mixture of 1-trans-(4-t-pentylcyclohexyl)-acetic acid (1.0 g) and silver nitrate (0.17 g) in acetonitrile (1.5 ml) was heated to reflux with vigorous stirring and diluted with ammonium persulfate (1.7 g). A solution of in water (3 ml) was added dropwise over a period of 1 hour. The mixture was heated at reflux for 1 hour, cooled on ice, and extracted with ether. The ether extract was washed with water, dried over magnesium sulfate and evaporated to give 2-[trans-(4't-pentylcyclohexyl)methyl]-3-chloro-1,4
-I got naphthoquinone. 700mg. Chlorquinone (700 mg) obtained above was dissolved in methanol (21 ml) and heated to reflux, and a solution containing potassium hydroxide (700 mg) in water (7 ml) was added dropwise over 10 minutes. After refluxing for an additional 15 minutes, the mixture was quickly cooled to room temperature and acidified with concentrated hydrochloric acid.
The mixture was extracted with chloroform and washed thoroughly with water. The organic extracts were dried over magnesium sulfate and evaporated under reduced pressure to an oil. Chromatographed on silica gel, eluted with toluene, recrystallized with acetonitrile, 2-[trans-(4'-t-pentylcyclohexyl)methyl]-3-hydroxy-1,4-
Obtained naphthoquinone. 150mg. Melting point 132-135℃. Example 4 An irrigation formulation for use in cattle may be prepared as follows. 2-[trans-(4-t-butylcyclohexyl)methyl]3-hydroxy-1,4-naphthoquinone 4 parts by weight Dimethyl sulfoxide 10 parts by weight Castor oil 100 parts by weight. Example 5 An aqueous suspension may be prepared as follows. 2-[trans-(4-t-butylcyclohexyl)methyl]3-hydroxy-1,4-naphthoquinone 1.00 parts by weight Neosyl 16.00 parts by weight Bentonite 3.20 parts by weight Glycerin 15.00 parts by weight Sodium benzoate 1.00 parts by weight Bevaloid35/2 1.00 parts by weight Example 6 The following finely divided materials can be mixed to form a salt block. 2-[trans-(4-t-butylcyclohexyl)methyl]-3-hydroxy-1,
4-Naphthoquinone (0.5 parts by weight) Sodium chloride (99.5 parts by weight) The mixture is pressed into blocks. Example 7 The following paste can be prepared. 2-[trans-(4-t-butylcyclohexyl)methyl]-3-hydroxy-1,4-naphthoquinone 3.0 parts by weight Gum tragacanth 4.0 parts by weight Bevaloid 35/3 1.0 parts by weight Nipagin “M” 0.1 parts by weight Glycerin 19.0 parts by weight Water 72.9 parts by weight 100.0 Example 8 A solution for hypodermic injection may be prepared by mixing as follows. 2-[trans-(4-t-butylcyclohexyl)methyl]3-hydroxy-1,4-naphthoquinone 4.5 parts by weight Methocel 2.0 parts by weight Nipagin “M” 0.1 parts by weight Water 93.4 parts by weight 100.0 Example 9 Mixed as follows. A solution for intramuscular injection may be prepared. 2-[trans-(4-t-butylcyclohexyl)methyl]-3-hydroxy-1,4-naphthoquinone 9.5 parts by weight Dimethyl sulfoxide 19.0 parts by weight Sorbitan monooleate 4.5 parts by weight Corn oil 67.0 parts by weight 100.0 Example 10 Next An injectable formulation was prepared. 2-[trans-(4-t-butylcyclohexyl)methyl]-3-hydroxy-1,4-naphthoquinone 5 parts N-methyl-pyrrolidone 48.3 parts Tween 80 2 parts Span 80 4.7 parts Miglyol 812 40 parts 100.0 Example 11 Inhydro-activity against T. parva and T. annulata Inhydro results showing the effect of compounds of formula () with R = CH ₃ were compared to the known anti-Theilerial agents 2-hydroxy-3-cyclohexyl-1,4-naphthoquinone and A comparison with 2-hydroxy-3-cyclohexylmethyl-1,4-naphthoquinone is also shown. Cultures of bovine gonobarblasts infected with the macroschizont stage of T. parva were treated with various concentrations of 2-[trans-(4-t-butylcyclohexyl)methyl]-3-hydroxy-1,4 -naphthoquinone, 2-hydroxy-3-cyclohexyl-1,4-naphthoquinone, or 2-hydroxy-3-cyclohexylmethyl-1,4-naphthoquinone for 48 hours.
The cells were cultured without any other drugs as a control. Compounds were tested in a 4-culture dilution series and ED50 values were determined. Each compound was replicated at least twice. ED50
The number of Schizont-infected cells in culture is 48
During incubation for hours, the drug dose (mg/) is reduced to 50% of the untreated control. The results are shown in Table 1. [Table] *: Example with no data 12 In vivo activity against T. parva The effect of 2-[trans-(4-t-butylcyclohexyl)methyl]-3-hydroxy-1,4-naphthoquinone was evaluated using known anti-Theileria agents. is 2
-Hydroxy-3-cyclohexyl-1,4-naphthoquinone. Purebred Friesian male calves weighing 180-250 kg and negative for serum antibodies to T. parva were given T.
1.0 ml of P. parva stabilizer 147 was inoculated subcutaneously. On the third day of distinct fever, 2-[trans-(4-t-butyl-cyclohexyl)-methyl]-3-hydroxy-
1,4-naphthoquinone and a similar formulation of 2-hydroxy-3-cyclohexyl-1,4-naphthoquinone were injected subcutaneously into each group of cows. Table 2 shows the recovery status compared to the control. [Table] Number [Table] Example 13 In vivo activity against T. annulata from 115
of T. annulata in a female Jersey calf weighing 140Kg.
Tick infected with Hissar strain ( Hyalomma
The infection was carried out by injection of 0.8 ml of stabilate prepared by homogenizing S. anatolicum (anatolicum anatolicum ).
2-[trans-(4-t-butylcyclohexyl)methyl]-3-hydroxy-1, 12 days after infection.
4-naphthoquinone (formulation of Example 9) was injected into the neck muscle. Table 3 shows the recovery status of each group and the control. [Table] Start of treatment
after
Continuous fever
number of days

Claims (1)

[Claims] 1 Formula () (wherein R is a C _1-10 alkyl group) or a pharmaceutically acceptable salt thereof, together with a carrier acceptable in the veterinary field, for the treatment or prevention of Theileriosis. formulations for use. 2. The preparation according to claim 1, wherein R in the compound of formula () is a straight-chain alkyl group. 3. The formulation according to claim 1 or 2, wherein R in the compound of formula () is a linear _C1-4 alkyl group. 4. The preparation according to any one of claims 1 to 3, wherein R in the compound of formula () is methyl. 5. The preparation according to any one of claims 1 to 3, wherein R in the compound of formula () is ethyl. 6. A formulation according to any one of claims 1 to 5, which is in the form of a cis/trans mixture of a compound of formula (). 7. The formulation according to claim 6, wherein the isomer ratio of the compound of formula () is about 1:1. 8. A formulation according to any one of claims 1 to 5, wherein the compound of formula () is in substantially pure trans isomer form. 9. A formulation according to any one of claims 1 to 5, wherein the compound of formula () is in substantially pure cis isomer form.