HK1235776A1 - Crystal of azole benzene derivative - Google Patents

Description

Crystal of azole benzene derivative

Technical Field

The present invention relates to a novel crystal of an azole (azole benzene) derivative which is effective as a therapeutic or prophylactic agent for diseases involving xanthine oxidase such as gout, hyperuricemia, tumor lysis syndrome, urethral calculus, hypertension, dyslipidemia, diabetes, arteriosclerosis, cardiovascular diseases such as cardiac insufficiency, renal diseases such as diabetic nephropathy, respiratory diseases such as chronic obstructive pulmonary disease, inflammatory bowel disease, and autoimmune diseases.

Background

Xanthine oxidase is an enzyme that catalyzes the conversion of hypoxanthine into xanthine and then into uric acid in nucleic acid metabolism.

In response to the action of xanthine oxidase, xanthine oxidase inhibitors lower the level of uric acid in blood by inhibiting the synthesis of uric acid. That is, xanthine oxidase inhibitors are effective for the treatment of hyperuricemia and various diseases caused by hyperuricemia. On the other hand, hyperuricemia continues to cause deposition of urate crystals, and as a result, gouty arthritis called gout, gout nodules, are caused as pathological conditions. In addition, hyperuricemia has also been highlighted as a factor of lifestyle-related diseases or metabolic syndrome associated with obesity, hypertension, dyslipidemia, diabetes, and the like, and it has recently been clarified by epidemiological investigation: hyperuricemia is a risk factor for renal damage, urinary calculi, cardiovascular diseases (compiled by the revised committee on the guidelines for gout and nucleic acid metabolism, "therapeutic guidelines for hyperuricemia and gout 2 nd edition", Medical Review society, 2010). Further, xanthine oxidase inhibitors are expected to be effective in the treatment of diseases in which active oxygen clusters are involved, for example, in the treatment of cardiovascular diseases based on the effect of improving vascular function, because of their active oxygen species inhibitory activity (circulation 2006; 114: 2508) -2516).

Allopurinol (allopurinol) and febuxostat (febuxostat) are clinically used as a therapeutic agent for hyperuricemia, but allopurinol is reported to have side effects such as Stevens-Johnson syndrome, toxic epidermal necrobiosis, liver damage, renal function damage and the like (Nippon Rinsho, 2003; 61, suppul.1: 197-.

As compounds having xanthine oxidase inhibitory activity, for example, 2-phenylthiazole derivatives have been reported (patent documents 1 to 3).

On the other hand, in patent documents 4 and 5, dithiazolecarboxylic acid derivatives having a benzene ring at the center are reported. Further, in patent document 6 and patent document 7, biphenyl thiazole carboxylic acid derivatives are reported.

Documents of the prior art

Patent document

Patent document 1: international publication No. 92/09279;

patent document 2: japanese laid-open patent publication No. 2002-105067;

patent document 3: international publication No. 96/31211;

patent document 4: international publication No. 2011/139886;

patent document 5: international publication No. 2011/101867;

patent document 6: international publication No. 2010/018458;

patent document 7: international publication No. 2010/128163.

Disclosure of Invention

Problems to be solved by the invention

The purpose of the present invention is to provide a crystal of a novel compound which is effective as a therapeutic or prophylactic agent for diseases involving xanthine oxidase, such as gout, hyperuricemia, tumor lysis syndrome, urinary calculi, hypertension, dyslipidemia, diabetes, cardiovascular diseases such as arteriosclerosis and cardiac insufficiency, renal diseases such as diabetic nephropathy, respiratory diseases such as chronic obstructive pulmonary disease, and inflammatory bowel disease and autoimmune diseases.

Means for solving the problems

The present inventors have intensively studied for the above-mentioned purpose and, as a result, found that: 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid (hereinafter, also referred to as compound (I)) can be crystallized, and at least 1 polymorph is present.

Namely, the present invention is as follows:

[1] crystals of 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid;

[2] the crystal of [1], wherein the crystal is a crystal;

[3][2]wherein in the powder X-ray diffraction spectrum, at a diffraction angle of 2θCharacteristic peaks at 7.2 °, 11.3 °, 15.9 °, 17.9 °, 20.8 °, 22.3 °, 23.1 °, 23.8 °, 24.3 °, and 28.6 °;

[4] [2] wherein the powder X-ray diffraction spectrum has the pattern shown in FIG. 1;

[5] [2] the crystal, wherein an exothermic peak in thermogravimetry/differential thermal analysis is 232 ℃;

[6] a pharmaceutical composition comprising the crystal of any one of [1] to [5] and a pharmaceutically acceptable carrier;

[7] a xanthine oxidase inhibitor containing a crystal of any one of [1] to [5] as an active ingredient; and

[8] a therapeutic or prophylactic agent for one or more diseases selected from gout, hyperuricemia, tumor lysis syndrome, urethral calculus, hypertension, dyslipidemia, diabetes, cardiovascular diseases, renal diseases, respiratory diseases, inflammatory bowel diseases and autoimmune diseases, which comprises the crystal according to any one of [1] to [5] as an active ingredient.

Effects of the invention

The present invention provides a crystal of an oxazolyl benzene derivative which is useful as a therapeutic or prophylactic agent for a disease in which xanthine oxidase is involved, such as gout, hyperuricemia, tumor lysis syndrome, urinary calculi, hypertension, dyslipidemia, diabetes, cardiovascular diseases such as arteriosclerosis or cardiac insufficiency, renal diseases such as diabetic nephropathy, respiratory diseases such as chronic obstructive pulmonary disease, inflammatory bowel disease or autoimmune diseases. The crystal can be used as raw material for preparing medicinal products.

Brief Description of Drawings

FIG. 1: fig. 1 is a powder X-ray diffraction spectrum of a crystal a.

Detailed Description

"xanthine oxidase" is generally used in the "broad sense" which means an enzyme catalyzing an oxidation reaction from hypoxanthine to xanthine and then to uric acid and in the "narrow sense" which means an oxidase type xanthine oxidoreductase which is one of enzymes catalyzing the same reaction, and in the present invention, unless otherwise specified, "xanthine oxidase" is a generic term for an enzyme catalyzing an oxidation reaction from hypoxanthine to xanthine and then to uric acid. In the xanthine oxidoreductase responsible for this reaction, there are two types, an oxidase type and a dehydrogenase type, which are also included in the xanthine oxidase of the present invention. In the terms of "xanthine oxidase inhibitory activity", "xanthine oxidase inhibitor" and the like, "xanthine oxidase" has the same meaning as defined above unless otherwise specified.

The crystal of the present invention is characterized in that: powder X-ray diffraction spectroscopy and/or thermogravimetry/differential thermal analysis (TG/DTA), and the like. The powder X-ray diffraction (XRD) spectrum of the crystal of the present invention shows a characteristic pattern having a characteristic diffraction angle 2θThe value of (c). In addition, the crystals of the present invention exhibit characteristic thermal behaviors even in thermogravimetry/differential thermal analysis (TG/DTA), respectively.

The A crystal of the present invention has a diffraction angle of 2 in a powder X-ray diffraction spectrumθ＝7.2°、11.3°、15.9°、Characteristic peaks are present at 17.9 °, 20.8 °, 22.3 °, 23.1 °, 23.8 °, 24.3 °, and 28.6 °. In addition, the a crystal of the present invention has a pattern shown in fig. 1 in a powder X-ray diffraction spectrum. In thermogravimetry/differential thermal analysis (TG/DTA), there is an exothermic peak at 232 ℃. The A crystal is an anhydrous crystal.

Here, the "characteristic peak" refers to a peak mainly recognized in a powder X-ray diffraction spectrum of each crystal polymorph and an intrinsic peak. The crystal identified (specified) by the diffraction angle of the present invention also contains a substance in which peaks other than the above-described characteristic peaks are observed.

Diffraction angle 2 in powder X-ray diffraction SpectrumθCan slightly vary depending on the measurement conditions, and therefore, even at 2θIn the case of slight differences, the identity of the crystal form should be determined by appropriately referring to the pattern of the entire spectrum, and crystals within the error range are also included in the present invention. As 2θFor example, +/-0.5 deg. and + -0.2 deg.. That is, the crystal identified by the diffraction angle also includes substances that are uniform within a range of ± 0.5 ° to ± 0.2 °.

In thermogravimetry/differential thermal analysis (TG/DTA), "exothermic peak" and "endothermic peak" are temperatures at the starting point of the peak, and refer to the starting temperatures of the exotherm and endotherm determined by extrapolation. Endothermic and exothermic peaks in TG/DTA may also vary slightly depending on the assay conditions. The error is, for example, in the range of. + -. 5 ℃ or. + -. 2 ℃. That is, the crystals identified by the above peaks also include substances that are consistent within the range of. + -. 5 ℃ to. + -. 2 ℃.

In addition, regarding any of the powder X-ray diffraction spectrum and TG/DTA, the difference between the measured value of each crystal obtained by, for example, the method described in the examples of the present application and the value described in the present application, which is a standard of the crystal, is also allowed as a measurement error. That is, within the error range calculated by the above method, a crystal having a diffraction angle or an endothermic and exothermic peak matching each other is also included in the crystal of the present invention.

Crystals A of 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid can be synthesized, for example, by the following method.

Synthesis of Compound (A-2)

[ chemical formula 1]

(in the formula, Y¹And Y²Represents a leaving group. )

As Y¹And Y²Examples of the leaving group include: halogen atom, methylsulfonyloxy group, p-toluenesulfonyloxy group, trifluoromethanesulfonyloxy group and the like. The reaction is carried out by reacting neopentyl alcohol with a leaving group Y in the compound (A-1) in the presence of a base²A method for synthesizing the compound (A-2) by a reaction. As the base used, there may be used: inorganic salts such as sodium hydride, sodium hydroxide, potassium hydroxide, lithium hydroxide, sodium carbonate, potassium carbonate, and cesium carbonate; metal alcoholates (alkoxide) such as sodium ethoxide, sodium methoxide and potassium tert-butoxide; triethylamine, pyridine, 4-aminopyridine, N-ethyl-N, N-Diisopropylamine (DIPEA), 1, 8-diazabicyclo [5.4.0 ]]Organic amines such as-7-undecene (DBU). The reaction is carried out by reacting an equivalent or slight excess of a base and an equivalent or excess of neopentyl alcohol with the compound (A-1) at 0 to 140 ℃ in a solvent inert to the reaction, and then adding the compound (A-1) thereto, usually for 0.5 to 16 hours. The reaction is preferably carried out under an inert gas atmosphere such as nitrogen. Here, the solvent is not particularly limited, and examples thereof include: aromatic hydrocarbons such as benzene, toluene, and xylene; ethers such as diethyl ether, Tetrahydrofuran (THF), 1, 4-dioxane, 1, 2-dimethoxyethane, and 1, 2-diethoxyethane; halogenated hydrocarbons such as dichloromethane, 1, 2-dichloroethane, and chloroform; n, N-Dimethylformamide (DMF), N-methylpyrrolidone (DMSO), water, or a mixed solvent thereof.

Synthesis of Compound (A-4)

[ chemical formula 2]

(wherein R represents an alkyl group having 1 to 6 carbon atoms.)

The present synthesis method is a method for synthesizing the compound (A-4) by coupling the compounds (A-2) and (A-3). As Y¹Examples of the leaving group include: halogen atom, methylsulfonyloxy group, p-toluenesulfonyloxy group, trifluoromethanesulfonyloxy group and the like. The reaction is carried out by using compounds (A-2) and (A-3) in an equal amount or an excess amount, adding a ligand, a carboxylic acid and a copper (valence I or II) salt as necessary in a solvent inert to the reaction in the presence of a base and a transition metal catalyst, and reacting at room temperature to reflux under heating for usually 0.5 hours to 2 days. The reaction is preferably carried out under an inert gas atmosphere such as nitrogen. Here, the solvent is not particularly limited, and examples thereof include: aromatic hydrocarbons such as benzene, toluene, and xylene; ethers such as diethyl ether, Tetrahydrofuran (THF), 1, 4-dioxane, 1, 2-dimethoxyethane, and 1, 2-diethoxyethane; halogenated hydrocarbons such as dichloromethane, 1, 2-dichloroethane, and chloroform; alcohols such as methanol, ethanol, 2-propanol, and butanol; n, N-Dimethylformamide (DMF), N-methylpyrrolidone (DMSO), water, or a mixed solvent thereof. Examples of the base include: lithium hydride, sodium hydride, potassium hydride, sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, cesium carbonate, potassium fluoride, cesium fluoride, tripotassium phosphate, sodium acetate, potassium acetate, etc., metal salts (lithium salt, sodium salt, potassium salt, magnesium salt) of alcoholate having 1 to 6 carbon atoms, metal salts (lithium salt, sodium salt, potassium salt, magnesium salt) of alkyl anion having 1 to 6 carbon atoms, tetra (alkyl) ammonium salt having 1 to 4 carbon atoms (fluoride salt, chloride salt, bromide salt), diisopropylethylamine, tributylamine, N-methylmorpholine, diazabicycloundecene, diazabicyclooctane, imidazole, etc. Examples of the transition metal catalyst include: copper, palladium, cobalt, iron, rhodium, ruthenium, iridium, and the like. Examples of the ligand include: III(tert-butyl) phosphine, tri (cyclohexyl) phosphine, tert-butyldicyclohexylphosphine, di (tert-butyl) cyclohexylphosphine, di (tert-butyl) methylphosphine, or the like. Examples of the copper (I-valent or II-valent) salt include: copper (I) chloride, copper (I) bromide, copper (I) iodide, copper (I) acetate, copper (II) fluoride, copper (II) chloride, copper (II) bromide, copper (II) iodide, copper (II) acetate, hydrates thereof, mixtures thereof, and the like. Examples of the carboxylic acid include: formic acid, acetic acid, propionic acid, n-butyric acid, isobutyric acid, valeric acid, isovaleric acid, pivalic acid, trifluoroacetic acid, and the like.

Synthesis of Compound (A-5)

[ chemical formula 3]

(wherein R represents an alkyl group having 1 to 6 carbon atoms.)

The synthesis method is a method for synthesizing the compound (A-5) by reducing the nitro group of the compound (A-4). The reaction is carried out by reacting the compound (A-4) in a solvent inert to the reaction, in the presence of a transition metal catalyst, under a hydrogen atmosphere at room temperature to under heating reflux, usually for 0.5 hour to 2 days. Here, the solvent is not particularly limited, and examples thereof include: aromatic hydrocarbons such as benzene, toluene, and xylene; ethers such as diethyl ether, Tetrahydrofuran (THF), 1, 4-dioxane, 1, 2-dimethoxyethane, and 1, 2-diethoxyethane; halogenated hydrocarbons such as dichloromethane, 1, 2-dichloroethane, and chloroform; alcohols such as methanol, ethanol, 2-propanol, and butanol; n, N-Dimethylformamide (DMF), N-methylpyrrolidone (DMSO), dimethyl sulfoxide (DMSO), ethyl acetate, or a mixed solvent thereof. As the transition metal catalyst, palladium-carbon, palladium hydroxide, palladium black, platinum-carbon, raney nickel (raneynickel) and the like are preferable.

Compound (A-6): synthesis of alkyl ester of 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid

[ chemical formula 4]

(wherein R and R¹Independently represents an alkyl group having 1 to 6 carbon atoms. )

The synthesis method is a method for synthesizing a tetrazole ring by reacting a compound (A-5), an orthoformate, and an azide compound. The reaction is carried out by using an equal or excess amount of the compound (A-5), orthoformate and azide in a solvent inert to the reaction in the presence of an acid at room temperature to under reflux with heating, usually for 0.5 hour to 2 days. The reaction is preferably carried out under an inert gas atmosphere such as nitrogen. Examples of orthoformates include: trimethyl orthoformate and triethyl orthoformate, and the like. Further, examples of the azide compound include: sodium azide, trimethylsilyl azide, and the like. Examples of the acid used include: organic acids such as formic acid and acetic acid; inorganic acids such as hydrochloric acid and sulfuric acid; lewis acids such as indium trifluoromethanesulfonate, ytterbium trifluoromethanesulfonate, zinc trifluoromethanesulfonate and indium trichloride. The solvent used in these reactions is not particularly limited, and examples thereof include: benzene, toluene, dichloromethane, dichloroethane, chloroform, carbon tetrachloride, diethyl ether, Tetrahydrofuran (THF), 1, 4-dioxane, 1, 2-dimethoxyethane, 1, 2-diethoxyethane, N-Dimethylformamide (DMF), N-methylpyrrolidone, Dimethylsulfoxide (DMSO), or a mixed solvent thereof, and an acid such as acetic acid may be used as the solvent.

Compound (I): synthesis of crystal A of 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid

[ chemical formula 5]

(wherein R represents an alkyl group having 1 to 6 carbon atoms.)

The crystal a of compound (I) can be prepared by a method comprising the steps of: a step of suspending the compound (A-7) in a solvent and adding an aqueous solution of a base to the suspension to hydrolyze the compound; and a step of neutralizing the reactant. The above method may further comprise a step of adding water to the neutralized product, followed by a step of stirring the reaction solution. Examples of the solvent for suspending compound (a-7) include: aromatic hydrocarbons such as benzene, toluene, and xylene; ethers such as diethyl ether, Tetrahydrofuran (THF), 1, 4-dioxane, 1, 2-dimethoxyethane, and 1, 2-diethoxyethane; halogenated hydrocarbons such as dichloromethane, 1, 2-dichloroethane, and chloroform; alcohols such as methanol, ethanol, 2-propanol, and butanol; n, N-Dimethylformamide (DMF), N-methylpyrrolidone (DMSO), water, or a mixed solvent thereof. Ethers, alcohols, water or a mixed solvent thereof is preferable.

In the compound (A-7), R is preferably an alkyl group having 1 to 6 carbon atoms, more preferably an ethyl group. Here, the alkyl group means a straight or branched aliphatic saturated hydrocarbon. Examples of the alkyl group having 1 to 6 carbon atoms include: methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, isopentyl, hexyl and the like are specific examples.

The hydrolysis reaction of compound (A-7) to compound (I) can be carried out by suspending compound (A-7) in the above-mentioned solvent (for example, 15 times the amount of compound (A-7)), and then reacting compound (A-7) with an equivalent amount or a slight excess of a base. Preferred bases include: sodium hydroxide, potassium hydroxide, lithium hydroxide. The reaction is carried out at 0-100 ℃, preferably 20-30 ℃. After the hydrolysis reaction, neutralization is carried out by reaction with an equivalent or slight excess of acid to the base used. A preferred acid is hydrochloric acid. The neutralization reaction can be carried out at 0-100 ℃, preferably 0-30 ℃.

Subsequently, water (for example, 5 times the amount of the compound (a-7)) was added to the neutralized reaction product, and after stirring for 1 hour, the precipitate was filtered off and dried to obtain crystals. The amount of solvent, the amount of water added, the stirring conditions, and the time until filtration (, respectively) are not particularly limited, but these conditions may affect the yield of crystals, chemical purity, particle size distribution, and the like, and therefore, it is preferable to combine them according to the purpose. The filtration may be carried out by a conventional method such as natural filtration, pressure filtration, reduced pressure filtration, or centrifugation. Drying may be carried out by a conventional method such as natural drying, drying under reduced pressure, drying by heating, and drying by heating under reduced pressure. The intermediate compound of the above reaction can be purified by conventional methods such as recrystallization, reprecipitation or various chromatographies during the synthesis, if necessary.

The crystals of the present invention can be distinguished by characteristic powder X-ray diffraction spectroscopy or thermogravimetry/differential thermal analysis (TG/DTA), but when other crystal forms exist, there is no concern about the incorporation rate thereof. When a specific crystal is obtained alone, incorporation is allowable to at least an extent undetectable by these measurement methods. In addition, when a specific crystal is used as a raw material as a drug, it is not allowable to contain another crystal.

The crystal of the present invention can be used as an active ingredient of a medicament. When other crystal forms exist, not only individual crystals but also a mixture of 2 or more kinds may be used.

In the present invention, by obtaining crystals of 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid, handling or reproducibility, stability, storage stability and the like at the time of production are more advantageous than those in an amorphous phase.

Pharmaceutical compositions can be prepared using the crystals of the invention and a pharmaceutically acceptable carrier.

The preparation containing the crystal of the present invention can be prepared by using additives generally used for the preparation. In the case of a solid preparation, examples of such additives include: lactose, white sugar, glucose, corn starch, potato starch, crystalline cellulose, light anhydrous silicic acid, synthetic aluminum silicate, magnesium metasilicate aluminate, calcium hydrogen phosphate and other excipients; binders (binders) such as crystalline cellulose, carboxymethyl cellulose, hydroxypropyl cellulose, sodium carboxymethyl cellulose, and polyvinyl pyrrolidone; disintegrants such as starch, sodium carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium and sodium carboxymethyl starch; lubricants such as talc and stearic acids; coating agents such as hydroxypropyl cellulose, hypromellose phthalate, and ethyl cellulose; in the case of a semisolid preparation, colorants may be mentioned: examples of the base such as white petrolatum in the case of a liquid preparation include: solvents such as ethanol; dissolving aids such as ethanol; preservatives such as parabens; isotonic agents such as glucose; buffers such as citric acid; antioxidants such as L-ascorbic acid; chelating agents such as EDTA; and suspending/emulsifying agents such as polysorbate 80.

The crystal of the present invention can be used in any dosage form such as a solid preparation, a semi-solid preparation, and a liquid preparation, and in any application preparation such as an oral preparation and a non-oral preparation (injection, transdermal preparation, eye drop, suppository, nasal preparation, inhalant, etc.).

A pharmaceutical composition containing the crystal of the present invention as an active ingredient can be used as a xanthine oxidase inhibitor; or a therapeutic agent or prophylactic agent for a disease in which xanthine oxidase is involved, such as gout, hyperuricemia, tumor lysis syndrome, urethral calculus, hypertension, dyslipidemia, diabetes, cardiovascular diseases such as arteriosclerosis or cardiac insufficiency, renal diseases such as diabetic nephropathy, respiratory diseases such as chronic obstructive pulmonary disease, inflammatory bowel disease or autoimmune disease. Herein, "prevention" means preventing a disease or onset in advance for an individual who has not yet suffered from the disease or onset, and "treatment" means curing, suppressing, or ameliorating the disease or symptoms for an individual who has suffered from the disease or onset.

Examples

[ measurement method ]

The powder X-ray diffraction of the crystal of the present invention was measured under the following conditions.

The device comprises the following steps: d8 DISCOVER With GADDS CS manufactured by Bruker AXS

Ray source: cu Ka; wavelength: 1.541838(10^-10m), 40kv-40mA, incident side flat-plate graphite monochromator and collimatorφ300 mu m two-dimensional PSPC detector, scanning 3-40 degrees.

Thermogravimetry/differential thermal analysis (TG/DTA) of the crystal of the present invention was measured using the following conditions.

The device comprises the following steps: TG8120 manufactured by Rigaku

Temperature rise rate: 10 ℃ per minute, ambient gas: nitrogen, sample pan: aluminum, control: alumina, sampling: 1.0 second, measurement temperature range: 25 to 300 DEG C

About measuring¹H NMR Spectroscopy (400MHz, DMSO-d)₆Or CDCl₃) The compound of (1), showing its chemical shift (: ppm) and coupling constant (J: hz). The following abbreviations respectively have the following meanings.

The device comprises the following steps: JMTC-400/54/SS manufactured by JEOL

s = singlet, d = doublet, t = triplet, q = quartet, brs = broad singlet, m = multiplet.

[ reference example 1]

2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl]-4-methyl-1, 3-thiazole-5- Preparation of ethyl formate

(1) After suspending 1.06g of neopentyl alcohol in 40.0mL of toluene and cooling to 0 ℃ under nitrogen, 1.35g of potassium tert-butoxide was added and stirring was carried out at 0 ℃ for 30 minutes. Next, 2.20g of 4-bromo-1-fluoro-2-nitrobenzene was added at 0 ℃ and the mixture was warmed to room temperature, followed by stirring at room temperature for 2 hours. Water was added to the reaction mixture, and extraction was performed with ethyl acetate. The organic layer was washed with brine, dried and concentrated under reduced pressure to obtain 3.12g of 4-bromo-1- (2, 2-dimethylpropoxy) -2-nitrobenzene.

(2) To 4.18g of 4-bromo-1- (2, 2-dimethylpropoxy) -2-nitrobenzene were added 3.04g of potassium hydrogencarbonate, 63mg of palladium (II) chloride and 297mg of copper (I) bromide, and the mixture was suspended in 45mL of toluene. Then, 2.97g of ethyl 4-methyl-1, 3-thiazole-5-carboxylate, 33. mu.L of isobutyric acid 1 and 333. mu.L of di-tert-butylcyclohexylphosphine were added, and the mixture was heated at 120 ℃ for 14 hours under a nitrogen atmosphere. The reaction mixture was subjected to celite filtration to remove insoluble matter, and water was added to the filtrate to conduct extraction with ethyl acetate. The organic layer was washed with brine, dried, concentrated under reduced pressure, and purified by a conventional method to obtain 5.13g of ethyl 2- [4- (2, 2-dimethylpropoxy) -3-nitrophenyl ] -4-methyl-1, 3-thiazole-5-carboxylate.

(3) 5.13gg of ethyl 2- [4- (2, 2-dimethylpropoxy) -3-nitrophenyl ] -4-methyl-1, 3-thiazole-5-carboxylate were suspended in 50mL of ethanol and, after addition of 500mg of palladium on carbon (10% wt), stirred under hydrogen at 50 ℃ for 20 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure, whereby 4.66g of ethyl 2- [ 3-amino-4- (2, 2-dimethylpropoxy) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylate was obtained.

(4) After suspending 2.58g of ethyl 2- [ 3-amino-4- (2, 2-dimethylpropoxy) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylate in 30mL of acetic acid, 962mg of sodium azide and 2.19g of triethyl orthoformate were added and the mixture was heated at 70 ℃ for 2 hours under a nitrogen atmosphere. After cooling to room temperature, 20mL of water was added to the reaction mixture and the mixture was purified by a conventional method to obtain 2.78g of ethyl 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylate.

[ example 1]

2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl]-4-methyl-1, 3-thiazole-5- Preparation of formic acid crystal A

2.58g of 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl]Ethyl (4-methyl-1, 3-thiazole-5-carboxylate) was dissolved in 30.0L of a mixed solution of tetrahydrofuran/methanol 1/1, and 6.50mL of a 2M aqueous sodium hydroxide solution was added thereto, followed by stirring at 20 to 30 ℃ for 3 hours. 6.50mL of 2M hydrochloric acid was slowly added to the reaction mixture while stirring at 20 to 30 ℃ and further 17.0mL of water was slowly added. The reaction solution was stirred at 20 to 30 ℃ for 1 hour, and then crystals were collected by filtration. The obtained crystal was washed with 7.0mL of a mixed solvent of methanol/water (1/1) and 7.0mL of water. The crystals were dried under reduced pressure at 50 ℃ to obtain 2.25g of 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl]-crystals of 4-methyl-1, 3-thiazole-5-carboxylic acid. The XRD of the resulting crystal is shown in fig. 1. At diffraction angle 2θPeaks were observed at 7.2 °, 11.3 °, 15.9 °, 17.9 °, 20.8 °, 22.3 °, 23.1 °, 23.8 °, 24.3 °, and 28.6 °. In addition, the exothermic peak in thermogravimetry/differential thermal analysis (TG/DTA) was 232 ℃.

[ reference example 2]

2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl]-4-methyl-1, 3-thiazole-5- Preparation of formic acid

307mg of ethyl 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylate was dissolved in 8.0mL of a mixed solution of tetrahydrofuran/methanol-1/1, and 1.0mL of a 2M aqueous sodium hydroxide solution was added thereto and stirred at room temperature for 3 hours. After 1.0mL of 2M hydrochloric acid was added to the reaction mixture and stirred, 6.0mL of water was added to purify the mixture by a conventional method, and 244mg of 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid was obtained.

[ reference example 3]

Measurement of xanthine oxidase inhibitory Activity

(1) Preparation of test Compound

2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid was dissolved in DMSO (manufactured by Sigma) to a concentration of 20mM, and then used after being adjusted to a target concentration.

(2) Measurement method

Evaluation of xanthine oxidase inhibitory activity of 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid was carried out by partially modifying the Method described in the literature (Method Enzymatic Analysis,1, 521-. This evaluation was carried out by measuring the activity of an oxidase type xanthine oxidoreductase. Specifically, a xanthine (manufactured by Sigma) solution prepared in advance to 10mM by a 20mM sodium hydroxide solution was prepared to 30. mu.M using a 100mM phosphate buffer, and 75. mu.L of the xanthine solution was added to a 96-well plate. The test compound diluted in DMSO to reach 100 times the final concentration was added at 1.5. mu.L per well, and after mixing, absorbance at 290nm was measured using a Microplate Reader (full-automatic quantitative plotter) SPECTRA MAX Plus 384 (manufactured by Molecular Devices). Next, an oxidase-type xanthine oxidoreductase (manufactured by Calbiochem corporation, buttermilk) was adjusted to 30.6mU/mL using 100mM phosphate buffer, and 73.5. mu.L of the enzyme was added to each well. The change in absorbance at 290nm was measured immediately after mixing for 5 minutes. The inhibitory rate of the test compound was calculated by assuming that the enzyme activity when DMSO was added instead of the test compound solution was 100%, and the inhibitory concentration of the test compound was calculated by fitting to a dose response curve, thereby calculating the 50% inhibitory concentration of the oxidase-type xanthine oxidoreductase.

2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl]4-methyl-1, 3-thiazole-5-carboxylic acid showed: IC of 1.0nM or less₅₀Xanthine oxidase inhibitory activity of < 5.0 nM.

[ reference example 4]

Uric acid lowering action in blood (Normal rat)

The blood uric acid-lowering effect of 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid was confirmed. The test compound suspended in a 0.5% methyl cellulose solution was forcibly administered to 8-9 weeks old male rats of Sprague-Dawley line (Charles River, Japan) using a feeding needle (feeding needle). Blood was collected from the tail vein 2 hours after administration, after which plasma was separated. Uric acid reduction rate was determined by the following equation by measuring the uric acid level in blood with an absorptiometer according to the uricase method using a uric acid measurement kit (L-type Wako UA. F: Wako pure chemical industries, Ltd.).

Uric acid reduction rate (%) (uric acid value of control animal-uric acid value of test compound administered to animal) x 100/uric acid value of control animal

2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid showed a uric acid lowering rate of 50% or more even at an amount of either 10mg/kg or 1 mg/kg.

The results show that: 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid has a potent blood uric acid lowering effect.

[ reference example 5]

Persistence of uric acid lowering action in blood (Normal rats)

2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid was administered to Sprague-Dawley line male rats in the same manner as in reference example 4. Blood was collected from the tail vein 24 hours after administration, after which plasma was isolated. Uric acid reduction rate was determined by the following equation by measuring the uric acid level in blood with an absorptiometer according to the uricase method using a uric acid measurement kit (L-type Wako UA. F: Wako pure chemical industries, Ltd.).

Uric acid reduction rate (%) (uric acid value of control animal-uric acid value of test compound administered to animal) x 100/uric acid value of control animal

2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid showed a uric acid lowering rate of 50% or more at an amount of 10mg/kg and 40% or more at an amount of 3mg/kg 24 hours after administration.

The results show that: 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid has a long-lasting blood uric acid lowering effect.

[ reference example 6]

Uric acid lowering action in blood (beagle dog)

A blood uric acid lowering effect in beagle dogs was confirmed for 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid. Beagle dogs (northern mountain labes) were forced to be orally administered test compounds suspended in a 0.5% methylcellulose solution. Blood was collected from the radial cutaneous vein at 8 hours after administration, after which plasma was isolated. The uric acid level in blood was measured by the LC-MS/MS method, and the uric acid lowering rate was determined by the following equation.

Uric acid reduction rate (%) (uric acid value of control animal-uric acid value of test compound administered to animal) x 100/uric acid value of control animal

2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid was administered at a dose of 10mg/kg at 8 hours after administration, showing a uric acid lowering rate of 80% or more.

The results show that: the compound of the present invention has a potent blood uric acid lowering effect on dogs

[ reference example 7]

Persistence of xanthine oxidase inhibitory Activity in tissues and plasma

In the present example, the "xanthine oxidase" of the present invention is referred to as "XO activity" and the "XOR activity" in order to distinguish the oxidation catalytic activity supported by the oxidase-type xanthine oxidoreductase from the oxidation catalytic activity supported by both the oxidase-type xanthine oxidoreductase and the dehydrogenase-type xanthine oxidoreductase. The "XO activity" and "XOR activity" also have the same meaning for "tissue XO activity", "plasma XO activity", "tissue XO inhibitory activity", "tissue XOR inhibitory activity", and the like. "tissue" includes liver, kidney, adipose tissue, intestinal tract, blood vessels. In this test, the XOR activity inhibition ratio and the XO activity inhibition ratio of the same sample were the same degree of value.

Tissue XO activity, tissue XOR activity and plasma XO activity were confirmed for 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid. The test compound suspended in a 0.5% methylcellulose solution was forcibly administered to 7-9 weeks old male rats of Sprague-Dawley line (Charles River, japan) using a feeding needle (feedingneedle). Blood and tissue draw from the abdominal aorta were performed 24 hours or 27 hours after administration. The resulting blood was centrifuged to collect plasma.

Tissue XO activity, tissue XOR activity, and plasma XO activity were measured by a reaction in which pterin was oxidized by each type of xanthine oxidoreductase to produce isoxapterin as a fluorescent substance. The tissues were homogenized in a potassium phosphate solution, ph7.4, containing 1mM EDTA and a protease inhibitor, to reach respective tissue concentrations to liver: 25mg/mL, kidney: 25mg/mL, fat: 5mg/mL, intestinal tract: 5mg/mL, blood vessel: 30mg/mL, and centrifuged at 12000rpm at 4 ℃ for 15 minutes. In the XO activity measurement, the supernatant or plasma of the tissue homogenate was mixed with a solution containing 50. mu.M pterin and reacted at 37 ℃. In the case of the XOR activity assay, the supernatant of the tissue homogenate was mixed with a solution containing 50. mu.M pterin and 50. mu.M methylene blue, and the reaction was carried out at 37 ℃. As a control, a solution containing an oxidase-type xanthine oxidoreductase (from buttermilk, Calbiochem Co., Ltd.) and 50. mu.M pterin was reacted by the same method. The fluorescence intensity of the produced isoxapterin was measured, corrected by the enzyme activity and protein concentration of the control, and calculated as the XO or XOR activity.

The XO inhibitory activity and the XOR inhibitory activity were determined by the following formulas.

XO inhibitory Activity (%). XO Activity or XOR Activity in control animals-XO Activity or XOR Activity in animals administered with test Compound). times.100/XO Activity or XOR Activity in control animals

Tissue and plasma XO inhibitory activity after about 27 hours of administration is shown in the table below.

[ Table 1]

2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid at a dose of 10mg/kg inhibited XO activity in the liver 27 hours after administration by more than 80% compared to control animals; XO activity in the kidney 27 hours after administration was more than 70% inhibited compared to control animals; XO activity in plasma 27 hours after administration was inhibited by more than 40% compared to control animals.

2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid at an amount of 1mg/kg inhibited XO activity in the liver 27 hours after administration by more than 80% compared to control animals; XO activity in the kidney 27 hours after administration was inhibited by more than 60% compared to control animals; XO activity in plasma 27 hours after administration was inhibited by more than 25% compared to control animals.

In addition, the tissue XOR inhibitory activity after 24 hours of administration is shown in the following table.

[ Table 2]

2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid at a dose of 10mg/kg inhibited the XOR activity in the liver 24 hours after administration by more than 80% compared to control animals; the XOR activity of the vessels 24 hours after administration was inhibited by more than 50% compared to control animals.

In addition, the XOR activity of the liver 24 hours after administration was inhibited by more than 80% compared to control animals at a dose of 1 mg/kg; the XOR activity of the vessels 24 hours after administration was inhibited by more than 30% compared to control animals.

The above results show that: the compounds of the present invention have long-term, sustained XO activity, XOR activity inhibition in various tissues.

Industrial applicability

The crystals of 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid of the present invention are useful as pharmaceuticals. Moreover, the crystal can be used as a raw material for the preparation of pharmaceuticals.

Claims (8)

1. Crystals of 2- [4- (2, 2-dimethylpropoxy) -3- (1H-1,2,3, 4-tetrazol-1-yl) phenyl ] -4-methyl-1, 3-thiazole-5-carboxylic acid.
2. 2. The crystal according to claim 1, wherein the crystal is a crystal.
3. 3. The crystal according to claim 2, wherein in the powder X-ray diffraction spectrum, at diffraction angle 2θ＝7.2°、11.3°、15.9°、17.9°、20.8°、22.3°、23.1°、23.8°Characteristic peaks at 24.3 °, and 28.6 °.
4. 4. The crystal of claim 2, wherein the powder X-ray diffraction spectrum has the pattern shown in fig. 1.
5. 5. The crystal of claim 2, wherein an exothermic peak in thermogravimetric/differential thermal analysis is 232 ℃.
6. 6. A pharmaceutical composition comprising the crystal of any one of claims 1 to 5 and a pharmaceutically acceptable carrier.
7. 7. A xanthine oxidase inhibitor comprising the crystal according to any one of claims 1 to 5 as an active ingredient.
8. 8. A therapeutic or prophylactic agent for one or more diseases selected from gout, hyperuricemia, tumor lysis syndrome, urethral calculus, hypertension, dyslipidemia, diabetes, cardiovascular diseases, renal diseases, respiratory diseases, inflammatory bowel diseases and autoimmune diseases, which comprises the crystal according to any one of claims 1 to 5 as an active ingredient.