CN104341362B - Triazole sulphonyl malonic acid compounds, Preparation Method And The Use - Google Patents

Abstract

The invention relates to the field of medicines related to hyperuricemia and gout. Specifically, the present invention relates to a class of uric acid transporter 1 inhibitors containing a triazole sulfonyl malonate structure, a preparation method thereof, a pharmaceutical composition containing them and their application in the preparation of diabetes medicines. Wherein, R is selected from H, C ₁ -C ₈ alkyl, and C ₃ -C ₆ cycloalkyl.

Description

Triazole sulfonyl malonate compound, 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 a triazolesulfonylmalonate structure that has 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 triazole sulfonyl malonate structure, 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.

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, R is selected from H, C ₁ -C ₈ alkyl, C ₃ -C ₆ cycloalkyl.

Preferably: R is selected from H, C ₁ -C ₄ alkyl, C ₃ -C ₄ cycloalkyl.

More preferred compounds of general formula (I) have the following structure,

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

Compound II reacts with dimethyl α-bromomalonate in the presence of a base to obtain compound IV; compound IV reacts with sodium nitrite and dichloroacetic acid in bromoform to obtain compound V; compound V is hydrolyzed under alkaline conditions Compound VI is obtained; compound VI is oxidized with 2 equivalents or more of an oxidant to obtain compound I.

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

5.65g (20mmol) of compound II-1 and 4.22g (20mmol) of compound III were dissolved in 100mL of dry DMF, stirred at room temperature, added 8.29g (60mmol) of solid K ₂ CO ₃ , and then the reaction mixture was stirred at room temperature until TLC tracking found that the reaction was complete (generally within 12h). The reaction mixture was poured into 400 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=435 ([M+Na] ⁺ ).

B. Synthesis of Compound V-1

5.23g (12mmol) compound IV-1 was dissolved in 30mL bromoform, stirred at room temperature, added 3.45g (50mmol) NaNO ₂ and 3.00g benzyltriethylammonium bromide, then added 6.45g (50mmol) dichloroacetic acid . The resulting mixture was stirred at room temperature until completion by TLC (typically within 12 h). The reaction mixture was poured into 300 mL ice water, stirred, extracted with 100 mL×3 CH ₂ Cl ₂ , the combined extracts were washed with 100 mL 2% Na ₂ S ₂ O ₃ solution and 100 mL 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 V-1, a white solid, ESI-MS, m/z=499 ([M+Na] ⁺ ).

C. Synthesis of Compound VI-1

3.81 g (8 mmol) of compound V-1 was dissolved in 30 mL of methanol, stirred at room temperature, 3 mL of 10% LiOH solution was added, and the resulting mixture was stirred at room temperature until the reaction was found to be complete by TLC tracking (generally within 3 h). The reaction mixture was poured into 200 mL of ice water, stirred, adjusted to pH=2-3 with concentrated hydrochloric acid, extracted with 100 mL×3 CH ₂ Cl ₂ , 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=446,448 ([MH] ^- ).

D. Synthesis of Compound I-1

2.69 g (6 mmol) of compound VI-1 was dissolved in 20 mL CH ₂ Cl ₂ , stirred, 2.40 g (14 mmol) m-chloroperoxybenzoic acid (mCPBA) was added, and stirred at room temperature for 5 h. The reaction mixture was poured into 200 mL ice water, stirred, extracted with 100 mL×3 CH ₂ Cl ₂ , the combined extract phases were washed successively with 100 mL 2% Na ₂ S ₂ O ₃ solution, 100 mL saturated NaHCO ₃ and 100 mL 5% brine, anhydrous sulfuric acid Sodium dry. 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 I-1, a white solid, ESI-MS, m/z=478, 480 ([MH] ^- )...

Example 2-6

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

Example 7

The IC50 values of the compounds of the present invention and related compounds on URAT1 inhibition were determined according to 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

compound _IC50 (hURAT1, nM) Lesinurad 22.4 I-1 13.1 I-2 32.8 I-3 25.75 --> I-4 28.2 I-5 21.5 I-6 16.7

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 (3)

1. The following compounds, 2. the method for compound described in synthetic claim 1: Compound II reacts with dimethyl α-bromomalonate in the presence of a base to obtain compound IV; compound IV reacts with sodium nitrite and dichloroacetic acid in bromoform to obtain compound V; compound V is hydrolyzed under alkaline conditions Compound VI is obtained; Compound VI is oxidized with 2 equivalents or more of an oxidant to obtain Compound I; wherein R is cyclopropyl or tert-butyl. 3. the application of the compound described in claim 1 in the preparation and treatment of hyperuricemia and gout medicine.