CN104262276B - Tetrazoleacetic acid compounds containing halogeno-benzene, Preparation Method And The Use - Google Patents

Abstract

Tetrazoleacetic acid compounds containing halogeno-benzene, Preparation Method And The Use, the present invention relates to the drug world relevant to hyperuricemia and gout。Specifically, the present invention relates to uric acid transporter body 1 inhibitor of the class tetrazoleacetic acid structure containing halogeno-benzene, its preparation method and pharmaceutical composition and their application in preparing diabetes medicament containing them。

Description

Tetrazolium acetic acid compound containing halogenated benzene, its preparation method and use

technical field

The invention relates to the field of drugs related to the treatment of hyperuricemia and gout. Specifically, the present invention relates to a class of uric acid transporter 1 (uratetransporter1, URAT1) inhibitors containing halobenzene tetrazoleacetic acid structures that have a therapeutic effect on hyperuricemia and gout, a preparation method, and a drug containing them Composition and use in medicine.

Background technique

Gout is a chronic metabolic disease characterized by hyperuricemia and pain caused by deposition of monosodium uric acid (MSU) in joints and other parts. The main reason is purine metabolism disorder and/or uric acid excretion disorder. It is estimated that there are more than 20 million gout patients worldwide. Drugs currently used to treat gout include anti-inflammatory drugs for pain relief (such as colchicine, etc.), drugs that inhibit uric acid production (xanthine oxidase inhibitors represented by allopurinol and febuxostat), crude uric acid Excretion drugs (uric acid excretion drugs represented by probenecid, besulfazone, benzbromarone, and losartan) and uricase (represented by pegloticase). These drugs have different degrees of toxic and side effects, such as benzbromarone has the risk of causing fulminant hepatitis, allopurinol has adverse reactions such as liver and bone marrow toxicity and allergic reactions, and so on.

Lesinurad (RDEA594) is an oral drug developed by Ardea that can inhibit the uric acid transporter (uratetransporter1, URAT1) in the kidney and excrete uric acid in the blood through the urine. It is currently in phase III clinical stage. Lesinurad was first developed from the antiviral drug RDEA806 developed by Valeant. The ownership of Lesinurad is currently owned by AstraZeneca due to the acquisition of the Ardea company.

The invention discloses a class of URAT1 inhibitors containing halogenated benzene tetrazoleacetic acid structures, and these compounds can be used to prepare medicines for treating hyperuricemia and gout.

Contents of the invention

One object of the present invention is to provide a class of compounds having general formula I with good activity.

Another object of the present invention is to provide a process for the preparation of compounds of general formula I.

Another object of the present invention is to provide a pharmaceutical composition containing a compound of general formula I as an active ingredient, and one or more pharmaceutically acceptable carriers, excipients or diluents, and its effect in the treatment of gout application.

The content of the present invention will now be specifically described in conjunction with the purpose of the present invention.

The compound of general formula (I) of the present invention has following structural formula:

Wherein, X is selected from halogen substituents.

Compounds of general formula (I) below are preferred,

Further, the following general formula (I) compounds are preferred,

The compound of general formula (I) of the present invention is synthesized by the following route:

Compound II reacts with 4-halobutyronitrile III in the presence of a base to obtain compound IV; compound IV reacts with dihalobenzene C ₆ H ₄ X ₂ (V) in the presence of a base to obtain compound VI; compound VI and NaN ₃ is reacted in the presence of NH ₄ Cl to obtain product VII; VII is reacted with compound 2-haloacetic acid VIII in the presence of a base to obtain product I; X is selected from Cl, Br and I, and the base is selected from organic bases and inorganic bases , and the definition of X is as described in claims 1-3.

The compound of the general formula I of the present invention has the inhibitory effect on URAT1, and can be used as an active ingredient for the preparation of therapeutic drugs for hyperuricemia and gout. The activity of the compound of general formula I in the present invention is verified by receptor binding assay.

The compounds of general formula I according to the invention are effective over a fairly wide dosage range. For example, the daily dose is about in the range of 1mg-500mg/person, divided into one or several administrations. The actual dosage of the compound of general formula I of the present invention can be determined by a doctor according to relevant conditions.

detailed description

The present invention will be further described below in conjunction with embodiment. It should be noted that the following examples are only for illustration, but not for limiting the present invention. Various changes made by those skilled in the art according to the teaching of the present invention shall be within the scope of protection required by the claims of the present application.

The synthesis of embodiment 1 compound I-1

A. Synthesis of Compound IV-1

3.66g (20mmol) of compound II-1 and 2.96g (20mmol) of compound III-1 were dissolved in 40mL of dry DMF, stirred, added 3.32g (20mmol) of potassium iodide and 6.91g (50mmol) of potassium carbonate, in a nitrogen atmosphere 100 React at °C until the reaction is complete (TLC tracking, generally 5h). After the reaction mixture was cooled, it was poured into 300 mL of ice water, stirred, extracted with 100 mL×3 CH ₂ Cl ₂ , the extract phases were combined, washed with 100 mL of 5% brine, and dried over anhydrous sodium sulfate. The desiccant was removed by suction filtration, the filtrate was evaporated to dryness on a rotary evaporator, and the obtained residue was purified by column chromatography to obtain compound IV-1, a white solid, ESI-MS, m/z=289 ([M+Na] ⁺ ).

B. Synthesis of Compound VI-1

3.19g (12mmol) of compound IV-1 and 1.76g (12mmol) of m-dichlorotoluene V-1 were dissolved in 30mL of dry DMF, stirred, and 4.98g (36mmol) of potassium carbonate was added to react at 100°C in a nitrogen atmosphere. until the reaction is complete. After the reaction mixture was cooled, it was poured into 300 mL of ice water, stirred, extracted with 100 mL×3 CH ₂ Cl ₂ , the extract phases were combined, washed with 100 mL of 5% brine, and dried over anhydrous sodium sulfate. The desiccant was removed by suction filtration, the filtrate was evaporated to dryness on a rotary evaporator, and the obtained residue was purified by column chromatography to obtain compound VI-1, a white solid, ESI-MS, m/z=399 ([M+Na] ⁺ ).

C. Synthesis of Compound VII-1

3.01g (8mmol) of compound VI-1 was dissolved in 20mL of dry DMF, stirred, 1.30g (20mmol) of NaN ₃ and 1.07g (20mmol) of ammonium chloride were added, and then reacted at 120°C in a nitrogen atmosphere until the reaction was complete . After the reaction mixture was cooled, it was poured into 200 mL of ice water, stirred, extracted with 100 mL×3 CH ₂ Cl ₂ , the extract phases were combined, washed with 100 mL of 5% brine, and dried over anhydrous sodium sulfate. The desiccant was removed by suction filtration, the filtrate was evaporated to dryness on a rotary evaporator, and the obtained residue was purified by column chromatography to obtain compound VII-1, a white solid, ESI-MS, m/z=442 ([M+Na] ⁺ ).

D. Synthesis of Compound I-1

2.09g (5mmol) of compound VII-1 and 1.39g (10mmol) of bromoacetic acid were dissolved in 15mL of ethanol, stirred at room temperature, and 3mL of aqueous solution containing 1.20g (30mmol) of NaOH was slowly added dropwise, and the resulting mixture was heated to 50°C. until the reaction is complete. After the reaction mixture was cooled, it was poured into 100 mL of ice water, stirred, adjusted to pH=2-3 with concentrated hydrochloric acid, extracted with 50 mL×3 CH ₂ Cl ₂ , the combined extracts were washed with 100 mL of 5% brine, and dried over anhydrous sodium sulfate. The desiccant was removed by suction filtration, the filtrate was evaporated to dryness on a rotary evaporator, and the obtained residue was purified by column chromatography to obtain compound IV-1 as a white solid, ESI-MS, m/z=476 ([MH] ^- ).

Example 2-11

Referring to the operation steps of Example 1, the compounds listed in the following table were prepared.

Example 12

The IC ₅₀ values of the compounds of the present invention and related compounds for URAT1 inhibition were determined by similar methods described in the literature (Example 12 in US2014/0005136).

Construction of a cell line stably expressing the humanized URAT1 transporter: The humanized URAT1 gene (SLC22A112) was subcloned from the plasmid pCMV6-XL-5 (Origene) into the eukaryotic expression plasmid pCMV6/neo (Origene). Gene sequencing confirmed that the humanized URAT1 was consistent with the information recorded in the gene bank (NM_144585.2). HEK293 human embryonic kidney cells (ATCC #CRL-1573) were cultured in EMEM tissue culture medium in an atmosphere of 5% _CO2 and 95% air. pCMV6/Neo/URAT1 was transfected onto HEK293 cells using L2000 type transfection reagent (Invitrogene). After 24 hours, the transfected cells were divided into tissue culture dishes with a diameter of 10 cm, continued to grow for one day, and then the medium was replaced with fresh medium containing 0.5 mg/mL G418 (Gibco). Eight days later, drug-resistant colonies were selected and collected, and tested for transport activity towards ¹⁴ C-labeled uric acid. HEK293/URAT1 cells were seeded on poly-D-lysine-coated 96-well plates at a density of 75,000/well.

These cells were grown overnight at 37° C. in an incubator, and then cooled to room temperature, and the culture medium was washed once with 250 μL/well of washing solution (125 mM sodium gluconate, 10 mM HEPES at pH=7.3). Add the compound to be tested or the blank control to a buffer containing 40 μM ¹⁴ C-labeled uric acid (54 mCi/mmol), which contains 125 mM sodium gluconate, 4.8 mM potassium gluconate, 1.2 mM potassium dihydrogen phosphate, 1.2mM magnesium sulfate, 1.3mM calcium gluconate, 5.6mM glucose, 25mM HEPES, final pH = 7.3. The 96-well plate was incubated at room temperature for 10 minutes, and then washed three times with 50 μL/well and 250 μL/well of the above cleaning solution. Microscint20 liquid flash agent was added to the 96-well plate, the plate was incubated overnight at 45°C, and then read on the TopCountPlateReader, and the IC ₅₀ was calculated accordingly.

The results are listed below.

IC ₅₀ values of some compounds of the present invention to URAT1

The above IC ₅₀ measurement results show that the compound of the present invention is a strong URAT1 inhibitor and can be used to prepare medicines for treating hyperuricemia and gout.

Claims (5)

1. have the compound of general formula I structure, Wherein, X is selected from halogen substituents. 2. The following compounds: 3. The compound of claim 2, selected from the group consisting of: 4. the method for the compound of general formula I defined in synthetic claim 1: Compound II reacts with 4-halobutyronitrile III in the presence of a base to obtain compound IV; compound IV reacts with dihalobenzene C ₆ H ₄ X ₂ (V) in the presence of a base to obtain compound VI; compound VI and NaN ₃ is reacted in the presence of NH ₄ Cl to obtain product VII; VII is reacted with compound 2-haloacetic acid VIII in the presence of a base to obtain product I; the base is selected from organic bases and inorganic bases, and the definition of X corresponds to claim 1 mentioned. 5. The application of the compound defined in any one of claims 1-3 in the preparation of medicines for the treatment of hyperuricemia and gout.