WO2021000527A1 - Tricyclic xor inhibitor, preparation method therefor and application thereof - Google Patents

Abstract

The present invention relates to the technical field of the pharmaceutical chemical industry, and disclosed are a tricyclic XOR inhibitor, a preparation method therefor and an application thereof. The structure of the tricyclic XOR inhibitor is represented by formula (I), and in the formula, B is imidazolyl, pyrazolyl, thiazolyl or triazolyl, and R is C1-C9 alkyl, alkoxy, amino or hydroxy, or R is cyano, halogen substituent, aldehyde group, carboxyl group, sulfonic acid group or nitro group. Using 5-bro-2-iodine-benzonitrile, R substituted aryl boronic acid and five-membered aromatic heterocyclic-4-ethylformate as starting raw materials, C-C coupling, C-N coupling and hydrolysis are successively performed to obtain a product. According to the present invention, the compound has completely different chemical structures from known XOR inhibitors, and has excellent inhibitory effects shown on XOR associated with gout so as to provide a new route for the preparation of anti-hyperuricemia or anti-gout drugs.

Description

Tricyclic XOR inhibitor and its preparation method and application Technical field

The invention belongs to the technical field of medicine and chemical engineering, and specifically relates to a tricyclic XOR inhibitor and a preparation method and application thereof.

Background technique

In recent years, with the rapid development of the global economy, people’s living standards have been greatly improved, accompanied by a rise in the prevalence of chronic diseases related to poor eating habits and lifestyles, among which gout is a serious impact The quality of life of patients is getting more and more attention and attention.

Gout is caused by the continuous increase of blood uric acid levels, resulting in the deposition of monosodium urate crystals in joints and other tissues. The level of blood uric acid is the core factor for the occurrence and development of gout. When the blood uric acid level exceeds the saturated dissolution amount in the blood, increased urate crystals are deposited in the joints, and a little crystal shedding can stimulate the surrounding tissues to produce inflammation. It causes redness, swelling, heat, and pain in the joints, and brings great pain and inconvenience to the patient's daily life. It can be seen that hyperuricemia and gout are closely related, and controlling the blood uric acid level is the key to preventing and treating the disease. Studies have found that hyperuricemia or gout is also closely related to the occurrence of hypertension, hyperlipidemia, atherosclerosis, diabetes and other diseases.

Decreased excretion or increased production of uric acid is the main cause of primary hyperuricemia. In the human body, uric acid has two important sources, exogenous and endogenous. Exogenous uric acid accounts for 20% of the source of uric acid. Almost all purines ingested from food are converted into uric acid in the body. Endogenous uric acid accounts for 80% of the source of uric acid, and the main reason for its increase is the deficiency of enzymes in purine metabolism, which affects the feedback regulation of purine metabolism and uric acid synthesis. Enzyme defects have the following manifestations: 1. Increased phosphoribose pyrophosphate synthase activity; 2. Increased phosphoribose pyrophosphate amidotransferase activity; 3. Decreased hypoxanthine-guanine phosphoribosyl transferase activity; 4. Yellow Purine oxidoreductase activity increased. On the other hand, because humans lack uric acid decomposing enzymes, uric acid cannot be metabolized into urea sacs and can only be excreted as a prototype, with the intestinal tract accounting for 30% and the kidney accounting for 70%. In the kidneys, the excreted uric acid is completely filtered by the glomeruli. 90% of the uric acid is reabsorbed by the renal proximal tubules, and only 10% of the uric acid is excreted in the urine. Recent studies have shown that a variety of uric acid transporters play an important role in the excretion of uric acid, including urate anion transporter (Universal Asynchronous Receiver/Transmitter, URAT1), uric acid transport related protein (Glucose transporter 9, GLUT9), The organic anion transporter family (OATs) family has the most important influence on the excretion of uric acid. In fact, a proper amount of uric acid plays a pivotal role in plasma. It can resist oxidation, inhibit the decomposition and synthesis of special enzymes, and can also chelate metal ions. Therefore, controlling the stability of blood uric acid levels in the body plays a vital role.

The treatment of gout is divided into the treatment of acute gout and the treatment of chronic gout. The standard treatment of acute gout is to reduce and control the inflammatory response, such as the use of colchicine, non-steroidal anti-inflammatory drugs, glucocorticoids (oral, intra-articular or intramuscular injection); inhibition of interleukin-1β (IL-1β) ) Of biological products such as anakinra, linacipr, kanazumab and so on. The treatment of chronic gout is mainly to control the blood uric acid level <6mg/dL through uric acid-lowering therapy. Common uric acid-lowering agents can be divided into three categories: xanthine oxidoreductase inhibitors (XORIs), such as allopurinol and non Busostat, topipristat; uric acid excretion drugs, such as probenecid, benzbromarone, and Recinade; uricase, such as polyethylene glycol recombinant uricase.

For a long time, the anti-gout drugs used in clinical practice have generally shown shortcomings such as poor efficacy, large side effects, and narrow applicability. However, emerging uricase drugs have also affected them due to immunogenicity, long-term safety and high price. widely used. XORIs has attracted much attention because of its clear mechanism of action, obvious curative effect and small side effects. Allopurinol (Allopurinol) is a kind of purine XORIs. It was marketed in the 1960s. It can be rapidly oxidized in the body to Oxypurinol, which has a stronger XOR inhibitory effect. It has also been marketed for use. Allopurinol can competitively inhibit the action of XOR and natural purine bases and block the production of uric acid, but it will produce many side effects, such as fever, allergic rash, diarrhea and abdominal pain. Febuxostat (Febuxostat) was developed by Teijin Pharmaceutical Company of Japan and was approved in the United States in 2008. It is the first non-purine XORIs approved by the US FDA for more than 40 years since the application of allopurinol. Its inhibitory ability is stronger than Allopurinol. Clinical studies have shown that febuxostat can significantly inhibit the production of uric acid, with small side effects and good safety, but it may still increase the risk of cardiovascular events. It has recently been warned by the FDA. Topiroxostat (Topiroxostat) is a new type of aryltriazole-based competitive XORIs developed by Fuji Yakuhin, Japan. It was approved for marketing in Japan in August 2013. Piraxostat (Y-700) is a type of mixed XORIs developed by Welfide. It has a strong and long-lasting effect on XOR to inhibit the production of uric acid. Y-700 is metabolized by the liver, and in vivo pharmacokinetic experiments show that it has good oral bioavailability. Degree, is still in the clinical research stage.

With the deepening of XOR research, people have found that inhibiting the activity of XOR can not only treat hyperuricemia, but also has a certain effect on ischemia, reperfusion injury, especially heart failure. XORIs with high efficiency and low toxicity have great development potential and Value. As far as gout is a stubborn disease, the design of new drugs with XOR as the target of action has received extensive attention, and a variety of highly active compounds have entered clinical trials, but they still face many problems such as large toxic and side effects, and more in-depth research is needed.

Summary of the invention

In view of the shortcomings and deficiencies of the above prior art, the primary purpose of the present invention is to provide a tricyclic XOR inhibitor.

Another object of the present invention is to provide a method for preparing the above tricyclic XOR inhibitor.

Another object of the present invention is to provide the application of the above-mentioned tricyclic XOR inhibitor in the preparation of uric acid or gout-lowering drugs.

The purpose of the present invention is achieved through the following technical solutions:

A tricyclic XOR inhibitor, its structure is shown in formula (I):

In formula (I), B is imidazolyl, pyrazolyl, thiazolyl or triazolyl;

R is an "electron-donating group" such as C1-C9 alkyl, alkoxy, amino or hydroxyl, or R is an "electron-withdrawing group" such as cyano, halogen substituent, aldehyde, carboxyl, sulfonic acid or nitro group".

More preferably, B is imidazolyl.

More preferably, R is a meta or para substitution; more preferably, it is a para substitution.

More preferably, R is a C1-C9 saturated aliphatic linear alkyl, branched alkyl or alkoxy group, or R is a cyano group, a nitro group; more preferably a tert-butyl group, a methoxy group or a cyano group.

More preferably, the structure of ring B is

More preferably, the tricyclic XOR inhibitor is a compound as described in any one of the following:

1-[4'-tert-butyl-2-cyano-(1,1'-biphenyl)]-1H-imidazole-4-carboxylic acid;

1-[4’-methoxy-2-cyano-(1,1’-biphenyl)]-1H-imidazole-4-carboxylic acid;

1-[3'-Methoxy-2-cyano-(1,1'-biphenyl)]-1H-imidazole-4-carboxylic acid.

The preparation method of the above tricyclic XOR inhibitor includes the following steps:

R取代苯基硼酸、无机碱、四三苯基磷钯加到有机溶剂中加热反应，得到

(1) Under the protection of argon,

R-substituted phenylboronic acid, inorganic base, and tetrakistriphenylphosphonium palladium are added to organic solvent and heated to react to obtain

CuI、无机碱、(E)-N’N-二甲基-1,2-环己二胺在有机溶剂中，经C-N偶联反应，得到

(2) Under the protection of argon,

CuI, inorganic base, (E)-N'N-dimethyl-1,2-cyclohexanediamine in organic solvent, through CN coupling reaction to obtain

经碱催化水解、酸化后得到

(3)

Obtained after alkali-catalyzed hydrolysis and acidification

The synthetic route of the above preparation method is shown in Figure 1.

Preferably, the organic solvent in steps (1) and (2) refers to DMF (N,N-dimethylformamide), and the inorganic base refers to K ₂ CO ₃ .

Preferably, the alkaline hydrolysis and acidification in step (3) refers to adding NaOH aqueous solution to the mixed solution (EtOH/THF) of ethanol and tetrahydrofuran to hydrolyze, and then adding HCl aqueous solution to acidify.

Application of the above-mentioned tricyclic XOR inhibitors in the preparation of drugs for lowering uric acid or gout.

Preferably, the uric acid or gout-lowering drug includes as an active ingredient a tricyclic XOR inhibitor or a pharmaceutically acceptable salt or ester thereof, and a pharmaceutically acceptable carrier.

Preferably, the pharmaceutically acceptable salt includes a salt formed by a tricyclic XOR inhibitor, a metal ion, an organic base, and a salt that can retain the biological effectiveness and properties of the parent compound.

More preferably, the metal ion is an alkali metal ion, an alkaline earth metal ion or an aluminum ion, and the organic base is ethanolamine, diethanolamine, triethanolamine, tromethamine, piperidine or piperazine.

The tricyclic XOR inhibitor of the present invention has the following advantages and beneficial effects:

The tricyclic XOR inhibitors of the present invention have a completely different chemical structure from the known XOR inhibitors, and are a new type of XOR inhibitors; as demonstrated in the following experimental examples, they are effective against gout-related yellow Purine oxidase shows an excellent inhibitory effect, and shows an excellent uric acid-lowering effect in acute and long-term hyperuricemia mouse models; therefore, they can be used to prevent and treat xanthine oxidase-related diseases, For example, hyperuricemia, heart failure, cardiovascular disease, hypertension, kidney disease, inflammation, arthropathy, etc.

Description of the drawings

Figure 1 is a synthetic route diagram of the tricyclic XOR inhibitor of the present invention.

Figure 2 is a graph showing changes in plasma uric acid content over time after administration of the compound D ₁ group of Example 1, the model control group, and the positive control group to acute hyperuricemia mice.

Fig. 3 is a graph showing the determination results of plasma uric acid, urea nitrogen and creatinine after administration of the compound D ₁ group of Example 1 and the normal control group, the model control group, and the positive control group to long-term hyperuricemia mice.

Detailed ways

The present invention will be further described in detail below in conjunction with the examples and drawings, but the implementation of the present invention is not limited thereto.

Example 1

Synthesis of 1-[4'-tert-butyl-2-cyano-(1,1'-biphenyl)]-1H-imidazole-4-carboxylic acid (D ₁ ):

(1) Under the protection of argon, add 5-bromo-2-iodo-benzonitrile (1.5g, 5mmol), tert-butylbenzeneboronic acid (1.3g, 7.5mmol), K ₂ CO ₃ (2.8g, 20mmol) , Tetraphenylphosphorylpalladium (0.3g, 0.3mmol) and DMF (6mL) were added to a 30mL two-necked flask, reacted at 120°C for 24h, TLC monitored the completion of the reaction, cooled to room temperature, and diluted with ethyl acetate 30mL, Add 30 mL of water, shake and separate, extract the aqueous phase with ethyl acetate (25 mL×3), combine the organic phases, wash with saturated brine (100 mL×2), dry with anhydrous magnesium sulfate, evaporate the solvent under reduced pressure, and purify by silica gel column (V _{ethyl acetate} :V _{petroleum ether} =1:20) 1.3 g of white solid 2-(4'-tert-butyl-phenyl)-5-bromo-benzonitrile (a ₁ ) was obtained, and the yield was 82.0%.

(2) Under argon protection, a ₁ (0.4g, 1.2mmol), ethyl imidazole-4-carboxylate (0.14g, 1.0mmol), CuI (19mg, 0.1mmol), K ₂ CO ₃ (0.3g, 2.1 mmol), and (E)-N'N-dimethyl-1,2-cyclohexyldiamine (28mg, 0.2mmol) and DMF (5mL) were added to a 25mL flask, and reacted at 110°C for 24h. Cool to room temperature, dilute with 15mL water, extract with ethyl acetate (15mL×3), wash with saturated brine (10mL), dry with anhydrous magnesium sulfate, evaporate the solvent under reduced pressure, and purify on silica gel column (V _{ethyl acetate} : V _{petroleum ether} =1:6) to obtain 0.1 g of white solid 1-[4'-tert-butyl-2-cyano-(1,1'-biphenyl)]-1H-imidazole-4-carboxylic acid ethyl ester (b ₁ ) , The yield is 52.0%.

(3) b ₁ (0.19g, 0.5mmol) is dissolved in a 1:1 volume ratio of tetrahydrofuran (4mL) and ethanol (4mL) mixed solution, add 1M NaOH 2mL, after the reaction is complete at 70 ℃, cool to room temperature , Add 1M HCl to adjust the pH to 2-3, add water to dilute, the solid precipitates, suction filtration, the filter cake is washed with water to neutral, and dried to obtain a white solid 1-[4'-tert-butyl-2-cyano-(1 ,1'-Biphenyl)]-1H-imidazole-4-carboxylic acid (D ₁ ) 0.17 g, the yield is 95.0%.

The structural characterization data of the product are as follows:

¹ H NMR(600MHz,DMSO-d ₆ )δ8.58(d,J=1.1Hz,1H,-NCH), 8.52(d,J=1.1Hz,1H,-NCH), 8.46(d,J=2.4 Hz, 1H, ArH), 8.18 (dd, J = 8.5, 2.4 Hz, 1H, ArH), 7.78 (d, J = 8.5 Hz, 1H, ArH), 7.60-7.56 (m, 4H, ArH), 1.35 ( s, 9H, -C ₆ H ₄ C(C H ₃ ) ₃ ). ¹³ C NMR (151MHz, DMSO-d ₆ ) δ163.80,152.10,143.68,137.30,136.01,135.39,134.42,132.02,128.89,126.13,126.07 ,125.94,124.38,118.31,111.74,34.96,31.60,31.50.

Example 2

Synthesis of 1-[4'-methoxy-2-cyano-(1,1'-biphenyl)]-1H-imidazole-4-carboxylic acid (D ₂ ):

(1) Under argon protection, 5-bromo-2-iodo-benzonitrile (1.5g, 5mmol), 4-methoxybenzeneboronic acid (1.1g, 7.5mmol), K ₂ CO ₃ (2.8g, 20mmol) ), palladium tetrakistriphenylphosphorus (0.3g, 0.3mmol) and DMF (6mL) were added to a 30mL two-necked flask, reacted at 120°C for 24h, TLC monitored the completion of the reaction, cooled to room temperature, and diluted with ethyl acetate 30mL Add 30mL water, shake and separate, the aqueous phase was extracted with ethyl acetate (25mL×3), the organic phases were combined, washed with saturated brine (100mL×2), dried over anhydrous magnesium sulfate, the solvent was evaporated under reduced pressure, silica gel column Purified (V _{ethyl acetate} :V _{petroleum ether} = 1:20) to obtain 1.1 g of white solid 2-(4'-methoxy-phenyl)-5-bromo-benzonitrile (a ₂ ), with a yield of 78.6% .

(2) Under argon protection, a ₂ (0.4g, 1.2mmol), imidazole-4-ethyl carboxylate (0.1g, 1.0mmol), CuI (19mg, 0.1mmol), K ₂ CO ₃ (0.3g, 2.1 mmol), (E)-N'N-dimethyl-1,2-cyclohexyldiamine (28mg, 0.2mmol) and DMF (5mL) were added to a 25mL flask and reacted at 110°C for 24h. Cool to room temperature, dilute with 15mL water, extract with ethyl acetate (15mL×3), wash with saturated brine (10mL), dry with anhydrous magnesium sulfate, evaporate the solvent under reduced pressure, and purify on silica gel column (V _{ethyl acetate} : V _{petroleum ether} =1:6) to obtain 0.1g of white solid 1-[4'-methoxy-2-cyano-(1,1'-biphenyl)]-1H-imidazole-4-carboxylic acid ethyl ester (b ₂ ) , The yield is 63.0%.

(3) b ₂ (0.17g, 0.5mmol) is dissolved in a 1:1 volume ratio of tetrahydrofuran (4mL) and ethanol (4mL) mixed solution, add 1M NaOH 2mL, after the reaction is complete at 70 ℃, cool to room temperature , Add 1M HCl to adjust the pH to 2-3, add water to dilute, the solid precipitates, suction filtration, the filter cake is washed with water to neutral, and dried to obtain a white solid 1-[4'-methoxy-2-cyano-(1 ,1'-Biphenyl)]-1H-imidazole-4-carboxylic acid (D ₂ ) 0.15 g, yield 96.5%.

The structural characterization data of the product are as follows:

¹ H NMR(600MHz,DMSO-d ₆ )δ8.56(s,1H,-NCH), 8.51(s,1H,-NCH), 8.44(d,J=2.3Hz,1H,ArH), 8.16(dd ,J=8.5,2.3Hz,1H,ArH),7.74(d,J=8.5Hz,1H,ArH),7.58(d,J=8.7Hz,2H,ArH),7.12(d,J=8.7Hz, 2H, ArH), 3.84 (s, 3H, -C ₆ H ₄ OC H ₃ ). ¹³ C NMR (151MHz, DMSO-d ₆ ) δ163.87,160.43,143.58,137.21,135.75,135.62,131.84,130.53,129.51, 125.95, 125.86, 124.23, 118.37, 114.78, 111.67, 55.81.

Example 3

Synthesis of 1-[3'-methoxy-2-cyano-(1,1'-biphenyl)]-1H-imidazole-4-carboxylic acid (D ₃ ):

(1) Under the protection of argon, 5-bromo-2-iodo-benzonitrile (1.5g, 5mmol), 3-methoxyphenylboronic acid (1.1g, 7.5mmol), K ₂ CO ₃ (2.8g, 20mmol) ), palladium tetrakistriphenylphosphorus (0.3g, 0.3mmol) and DMF (6mL) were added to a 30mL two-necked flask, reacted at 120°C for 24h, TLC monitored the completion of the reaction, cooled to room temperature, and diluted with ethyl acetate 30mL After that, 30 mL of water was added, and the layers were shaken. The aqueous phase was extracted with ethyl acetate (25 mL×3). The organic phases were combined, washed with saturated brine (100 mL×2), dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure. Column purification (V _{ethyl acetate} :V _{petroleum ether} = 1:20) to obtain 1.0 g of white solid 2-(3'-methoxy-phenyl)-5-bromo-benzonitrile (a ₃ ), the yield is 68.5 %.

(2) Under argon protection, a ₃ (0.4g, 1.2mmol), ethyl imidazole-4-carboxylate (0.1g, 1.0mmol), CuI (19mg, 0.1mmol), K ₂ CO ₃ (0.3g, 2.1 mmol), (E)-N'N-dimethyl-1,2-cyclohexyldiamine (28mg, 0.2mmol) and DMF (5mL) were added to a 25mL flask and reacted at 110°C for 24h. Cool to room temperature, dilute with 15mL water, extract with ethyl acetate (15mL×3), wash with saturated brine (10mL), dry with anhydrous magnesium sulfate, evaporate the solvent under reduced pressure, and purify on silica gel column (V _{ethyl acetate} : V _{petroleum ether} =1:6) to obtain 0.1 g of white solid 1-[3'-methoxy-2-cyano-(1,1'-biphenyl)]-1H-imidazole-4-carboxylic acid ethyl ester (b ₃ ) , The yield is 63.0%.

(3) b ₃ (0.17g, 0.5mmol) is dissolved in a 1:1 volume ratio of tetrahydrofuran (4mL) and ethanol (4mL) mixed solution, add 1M NaOH 2mL, after the reaction is complete at 70 ℃, cool to At room temperature, add 1M HCl to adjust the pH to 2-3, add water to dilute, solid precipitate, suction filtration, the filter cake is washed with water to neutral, and dried to obtain white solid 1-[3'-methoxy-2-cyano-( 1,1'-biphenyl)]-1H-imidazole-4-carboxylic acid (D ₃ ) 0.14 g, yield 90.0%.

The structural characterization data of the product are as follows:

¹ H NMR(600MHz,DMSO-d ₆ )δ8.61(d,J=4.7Hz,2H,-NCH), 8.49(d,J=1.8Hz,1H,ArH), 8.20(dd,J=8.4, 1.9Hz,1H,ArH),7.81(d,J=8.5Hz,1H,ArH),δ7.48(t,J=7.9Hz,1H,ArH),7.19(d,J=7.2Hz,1H,ArH) ), 7.11-7.08 (m, 1H, ArH), 3.84 (s, 2H, -C ₆ H ₄ OC H ₃ ). ¹³ C NMR (151MHz, DMSO-d ₆ ) δ 163.60, 159.84, 143.70, 138.60, 137.35, 136.17,135.04,132.07,130.45,126.04,125.89,124.43,121.41,118.11,115.14,114.76,112.02,55.77.

Activity evaluation of the products obtained in the above examples:

1. Evaluation of the in vitro inhibitory activity of compound D ₁ -D ₃ on XOR

(1) Solution preparation

Buffer: Dilute 10×PBS (pH 7.4) to 1×PBS. Unless otherwise specified, the PBS in the reaction system refers to 1×PBS.

Substrate: Weigh 15.2 mg of xanthine, add 45 mL of PBS to ultrasonically dissolve, and then add PBS to make the volume to 200 mL to obtain a substrate solution of 0.5 mmol/L.

Enzyme solution: Under ice bath, dilute 10.2μL xanthine oxidoreductase mother solution with 20mL PBS to obtain 0.5μg/100μL enzyme solution.

Compounds to be tested: Weigh the products D ₁ -D ₃ in Examples 1 to 3 accurately, prepare a 1mmol/L solution with DMSO, and store them at 20°C in the dark. Dilute with PBS to the required concentration before use, and control the DMSO content within 5% to ensure that it has no effect on enzyme activity.

(2) Determination

Add the above-prepared PBS solution, sample or blank solution (the blank solution is PBS solution), and 100 μL of enzyme solution in turn into the 96-well plate, incubate in the microplate reader at 37°C for 3 minutes, and then add to the incubated microplate The substrate initiates the reaction. Reading at 295nm is performed every 1 minute for a total of 5 minutes. Each experiment is tested in parallel three times. The initial velocity of the test compound at each concentration was converted into the percentage (%) of the inhibition rate based on the initial velocity in the absence of inhibitor, and the IC ₅₀ value was calculated. The results are shown in Table 1.

Table 1 Inhibitory activity of compound D ₁ -D ₃ on XOR (n=3)

From the results in Table 1, it can be seen that the compound D ₁ -D ₃ obtained in the present invention has the following structure-activity relationship: _1. The inhibitory activity of compound D ₁ is the strongest (IC ₅₀ =7.2 nM), which is equivalent to febuxostat (P>0.05) ); 2. The benzene rings of compounds D ₁ , D ₂ , and D ₃ are substituted with 4-tert-butylphenyl, 4-methoxyphenyl, and 3-methoxyphenyl, respectively, and their inhibitory activity decreases sequentially , Suggesting that alkyl substitution on benzene ring is better than alkoxy substitution. In addition, the inhibitory activity of compound D ₂ is about twice that of compound D ₃ , suggesting that para-substitution on the benzene ring is better than meta-substitution.

2. Evaluation of uric acid-lowering activity of compound D ₁ in a mouse model of acute hyperuricemia

After 18-22g SPF grade ICR mice were adaptively fed for one week, they were randomly divided into a model control group (potassium oxazinate 250 mg·kg ^-1 + hypoxanthine 400 mg·kg ^-1 ) and a positive control group (oxazine Potassium 250mg·kg ^-1 + Hypoxanthine 400mg·kg ^-1 + Febuxostat 5mg·kg ^-1 ), group D ₁ (potassium oxazine 250mg·kg ^-1 + hypoxanthine 400mg·kg ^-1 +D ₁ compound 5 mg·kg ^-1 ), 8 rats in each group.

The mice were weighed before the experiment, and the model control group and each administration group were given a subcutaneous injection of 250 mg·kg ^{-1 of} potassium oxazinate + intraperitoneal injection of 400 mg·kg ^{-1 of} hypoxanthine. After ¹ hour, the blood uric acid level was measured (recorded as 1h). Blood uric acid), and immediately give each drug group the drug to be tested. The model control group is given the same amount of solvent. 1h, 2h, 3h, 4h, 5h, 6h, 7h after administration, the uric acid value is measured (respectively Recorded as 2h, 3h, 4h, 5h, 6h, 7h, 8h blood uric acid). The results obtained are plotted with graphpad 6.0 and statistically analyzed by one-tailed Students t-test. The results are shown in Figure 2 (*P<0.05 compared with model control group, **P<0.01 compared with model control group).

It can be seen from Figure 2 that within 8 hours after administration, compound D ₁ can significantly reduce the blood uric acid level of mice with acute hyperuricemia.

3. Evaluation of uric acid-lowering activity of compound D ₁ in long-term hyperuricemia mouse model

After 18-22g SPF grade ICR mice were adaptively fed for one week, they were randomly divided into normal control group (normal saline) and model control group (potassium oxazine 250mg·kg ^-1 + hypoxanthine 150mg·kg ^-1 ), positive control group (potassium oxazine 250 mg·kg ^-1 + hypoxanthine 150 mg·kg ^-1 + febuxostat 5 mg·kg ^-1 ), group D ₁ (potassium oxazine 250 mg·kg ^-1 + Hypoxanthine 150 mg·kg ^-1 + D ₁ compound 5 mg·kg ^-1 ), 8 rats in each group.

The mice were weighed before the experiment, and the model control group and each administration group were given subcutaneous injection of 250 mg·kg ^{-1 of} potassium oxonate + intraperitoneal injection of hypoxanthine 150 mg·kg ^-1 at 9:00 every day, and the normal control group was given The same amount of normal saline, the blood uric acid level (recorded as 1h blood uric acid) was measured after 1 hour, and the drug to be tested was administered to each drug group immediately, and the normal control group and the model control group were given the same amount of solvent by gavage, repeated continuously The above experiment was 7 days. On the seventh day after the treatment drug was administered for 1 hour, the eyeballs were taken immediately, and the blood was collected at 0-4°C for 10 minutes. The blood was centrifuged at 10000 r/min for 5 minutes. The upper plasma was carefully drawn and used with an automatic biochemical analyzer (Beckman Coulter, AU5811, Tokyo). , Japan) Measure the values of Uric acid, Urea Nitrogen and Crea in plasma. The results are plotted in graphpad 6.0 to two-tailed Student's t-test statistical analysis, the results as shown in ^(# 3 as compared with the normal group, P <0.05, ^## as compared with the normal group, P <0.01, * and Model P<0.05 compared with group, **P<0.01 compared with model group).

It can be seen from Figure 3 that the blood uric acid level of the model control group was significantly higher than that of the normal group. The model was successfully established (P<0.001). The blood creatinine and urea nitrogen levels of the model control group were significantly higher than those of the normal group, suggesting long-term hyperuricemia It will cause some damage to the kidney function of mice (P<0.05). Compared with the model control group, both D ₁ and febuxostat have a significant effect of lowering uric acid on hyperuricemia mice (P<0.05). In addition, compound D ₁ shows a certain effect of lowering urea nitrogen and creatinine (P<0.05), indicating that compound D _{1 can not} only reduce blood uric acid activity, but also improve renal function damage to a certain extent. Therefore, based on the results of compound D ₁ uric acid lowering activity evaluation of compound D ₁ pending further study, and is expected to become a new type of uric acid lowering drugs.

The above-mentioned embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited by the above-mentioned embodiments, and any other changes, modifications, substitutions, combinations, etc. made without departing from the spirit and principle of the present invention Simplified, all should be equivalent replacement methods, and they are all included in the protection scope of the present invention.

Claims (10)

A tricyclic XOR inhibitor, characterized in that: the structure of the tricyclic XOR inhibitor is shown in formula (I):

In formula (I), B is imidazolyl, pyrazolyl, thiazolyl or triazolyl; R is a C1-C9 alkyl group, alkoxy group, amino group or hydroxyl group, or R is a cyano group, a halogen substituent, an aldehyde group, a carboxyl group, a sulfonic acid group or a nitro group. The tricyclic XOR inhibitor of claim 1, wherein B is imidazolyl. The tricyclic XOR inhibitor according to claim 1, characterized in that: R is meta or para substitution; R is saturated aliphatic linear alkyl, branched alkyl or alkoxy of C1～C9 , Or R is cyano, nitro. The tricyclic XOR inhibitor of claim 1, wherein the structure of the B ring is

The tricyclic XOR inhibitor of claim 1, wherein the tricyclic XOR inhibitor is a compound according to any one of the following: 1-[4'-tert-butyl-2-cyano-(1,1'-biphenyl)]-1H-imidazole-4-carboxylic acid; 1-[4’-methoxy-2-cyano-(1,1’-biphenyl)]-1H-imidazole-4-carboxylic acid; 1-[3'-Methoxy-2-cyano-(1,1'-biphenyl)]-1H-imidazole-4-carboxylic acid. The preparation method of a tricyclic XOR inhibitor according to any one of claims 1 to 5, characterized by comprising the following steps: (1) Under the protection of argon,

R-substituted phenylboronic acid, inorganic base, and tetrakistriphenylphosphonium palladium are added to organic solvent and heated to react to obtain

(2) Under the protection of argon,

CuI, inorganic base, (E)-N'N-dimethyl-1,2-cyclohexanediamine in organic solvent, through CN coupling reaction to obtain

(3)

Obtained after alkali-catalyzed hydrolysis and acidification

The method for preparing a tricyclic XOR inhibitor according to claim 6, wherein the organic solvent in steps (1) and (2) refers to DMF, and the inorganic base refers to K ₂ CO ₃ . The method for preparing a tricyclic XOR inhibitor according to claim 6, characterized in that: the alkaline hydrolysis and acidification in step (3) refer to adding NaOH aqueous solution to the mixed solution of ethanol and tetrahydrofuran to hydrolyze, and then Add HCl aqueous solution to acidify. Use of a tricyclic XOR inhibitor according to any one of claims 1 to 5 in the preparation of uric acid or gout-lowering drugs. The use of a tricyclic XOR inhibitor in the preparation of a uric acid or gout-lowering drug according to claim 9, characterized in that: the uric acid or gout-lowering drug comprises a tricyclic XOR inhibitor or its Pharmaceutically acceptable salts, esters and pharmaceutically acceptable carriers; the pharmaceutically acceptable salts include salts formed by tricyclic XOR inhibitors and metal ions or organic bases; the metal ions are alkali metal ions, Alkaline earth metal ion or aluminum ion, the organic base is ethanolamine, diethanolamine, triethanolamine, tromethamine, piperidine or piperazine.

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