CN117567406A - URAT1 inhibitors, compositions and uses thereof - Google Patents

Abstract

The present invention relates to the field of medicaments related to hyperuricemia and gout. In particular, the present invention relates to the use as a novel URAT1 inhibitor, compositions and uses thereof, in particular as a therapeutic agent for conditions associated with abnormal uric acid levels.

Description

URAT1 inhibitors, compositions and uses thereof

Technical Field

The invention belongs to the field of pharmaceutical chemistry, and particularly relates to quinoline derivatives for inhibiting URAT1, a composition thereof and a pharmaceutical application for treating or preventing diseases related to abnormal uric acid level.

Background

The disturbance of purine metabolism and/or reduced uric acid excretion at gout causes elevated blood uric acid, and repeated attacks of inflammatory diseases caused by deposition of urate crystals in articular synovium, bursa, cartilage and other tissues. Important biochemical basis for gout in hyperuricemia. The current common therapeutic drugs mainly comprise xanthine oxidase inhibitors (such as allopurinol, febuxostat and the like) for reducing uric acid generation, uric acid excretion-promoting drugs (such as propane sulfonic acid, fenofibrate (non-indication drug), losartan (non-indication drug), benzbromarone and the like) for promoting uric acid excretion.

Lesinurad is a selective uric acid reabsorption inhibitor developed by Aspirin, and can reduce uric acid secretion and reduce uric acid content in serum by inhibiting URAT1 transporter in kidney, reduce uric acid production and increase uric acid excretion. The drug was approved by the FDA for the treatment of high blood uric acid (hyperuricemia) associated with gout in month 12 of 2015, and was approved by the European Medicines Agency (EMA) for 18 of month 2 of 2016.

The invention discloses URAT1 inhibitors, which can be used for preparing medicines for treating hyperuricemia and gout.

Disclosure of Invention

The present invention provides URAT1 inhibitor compounds and methods of making the same. The invention also provides a pharmaceutical composition containing one or more compounds of the invention and application thereof in the aspects of treating hyperuricemia and gout.

To achieve the above object, the present invention provides, in one aspect, a compound as shown below and pharmaceutically acceptable salts thereof:

the invention also provides a pharmaceutical composition comprising a compound according to any one of the above, and pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers.

The invention also provides application of the compound and pharmaceutically acceptable salts thereof in preparing medicines for inhibiting URAT 1.

As a preferred embodiment, the use is in the manufacture of a medicament for the treatment or prevention of a disorder associated with abnormal uric acid levels.

The compound prepared by the invention has excellent URAT1 target inhibition effect, high selectivity and unexpected good activity.

Detailed Description

The invention is further illustrated by the following examples, which are given by way of illustration only and are not to be construed as limiting the scope of the invention in any way.

EXAMPLE 1 preparation of 3- (5- (4-bromonaphthalen-1-yl) furan-2-yl) propionic acid

Step 1: synthesis of Compounds 1-2

Naphthalene (10 mmol, 1) was added to a 250ml three-necked flask28 g), 50ml of dichloromethane, cooled to-30 ℃, bromine (30 mmol,4.79 g) is slowly added dropwise, the reaction is carried out at-30 ℃ to-25 ℃ in the dark, and TLC detects the disappearance of the raw material points. Ending the reaction with saturated NaHSO ₃ Quenching with aqueous solution, the organic layer was quenched with saturated NaHSO ₃ Aqueous washing (40 ml. Times.3), 2M aqueous sodium hydroxide washing (40 ml. Times.3), drying over anhydrous sodium sulfate, concentrating the organic solvent and purifying by column chromatography gave compound 1-2 (8 mmol,2.27g, 80%) as a white powder. ESI-MS, M/z=285 ([ m+h)] ⁺ )。

Step 2: synthesis of Compounds 1-3

Into a 250ml three-necked flask, compound 1-2 (10 mmol,2.84 g) and THF 50ml were added, n-butyllithium (11 mmol,1.6M,6.88 ml) was slowly added dropwise under nitrogen protection at-78℃and reacted at-78℃for 2 hours, then 5ml of THF solution of triisopropyl borate (15 mmol,2.82 g) was slowly added dropwise to the reaction system, and after the addition was completed, the mixture was slowly warmed to room temperature and stirred overnight, and after the TLC detection of the disappearance of the starting material, the reaction was completed. Dilute hydrochloric acid (20%, 20 ml) was added to quench, stirred at room temperature for 3 hours, then extracted with ethyl acetate (50 ml. Times.3), the organic phases were combined, washed with saturated brine (100 ml. Times.3), dried over anhydrous sodium sulfate, and the organic phase was concentrated to give compound 1-3 (7.6 mmol,1.90g, 76%) after purification by column chromatography. ESI-MS, M/z=251 ([ m+h)] ⁺ )。

Step 3: synthesis of Compounds 1-5

2, 5-dibromofuran (10 mmol,2.24 g), 50ml THF, and n-BuLi (11 mmol,6.88ml,1.6 mol/L) were slowly added dropwise under nitrogen protection at-78deg.C, and the reaction mixture was stirred at-78deg.C for 1 hour, then methyl bromopropionate (11 mmol,1.84 g) was slowly added, and the reaction was carried out at-78deg.C for 2 hours. After completion of the reaction, cooling to room temperature, quenched by addition of water, extracted with ethyl acetate (50 ml. Times.3), the combined organic layers were washed with saturated brine (100 ml. Times.3), dried over anhydrous sodium sulfate, concentrated in organic solvent, and purified by column chromatography to give compounds 1-5 (7.19 mmol,1.67g, 72%). ESI-MS, M/z=233 ([ m+h)] ⁺ )。

Step 4: synthesis of Compounds 1-6

Into a 250ml three-necked flask were charged the compounds 1-3 (12 mmol,3.00 g), 2- (5-bromofuran)-2-yl) -acetic acid methyl ester (10 mmol,2.32 g), potassium carbonate (10 mmol,1.38 g), 80% aqueous ethanol 50ml, pd (dba) 2 (0.10 mmol,0.06 g), at 50℃and TLC detected the end of the reaction. The organic solvent was concentrated, cooled to room temperature, 30ml of water was added, extracted with ethyl acetate (50 ml x 3), the organic layers were combined, washed with saturated brine (100 ml x 3), dried over anhydrous sodium sulfate, and purified by column chromatography to give compounds 1-6 (6.0 mmol,2.15g, 60%). ESI-MS, M/z=359 ([ m+h)] ⁺ )。

Step 5: synthesis of Compound 1

Adding compound 2-6 (10 mmol,3.58 g), 30ml methanol, adding 2N sodium hydroxide aqueous solution 6ml under ice water bath, returning to room temperature, stirring, TLC detecting reaction completion, concentrating organic solvent, adding 30ml water, extracting with ethyl acetate (30 ml×3), discarding organic layer, and regulating pH of water layer with 10% hydrochloric acid aqueous solution<2, ethyl acetate extraction (50 ml x 3), washing with saturated brine (100 ml x 3), drying over anhydrous sodium sulfate, and concentrating the organic solvent to give compound 1 (9.4 mmol,3.23g, 94%). ESI-MS, M/z=345 ([ m+h)] ⁺ )。

The compounds listed in the following table were prepared with reference to the preparation method of example 1:

experimental example

The IC50 values of the compounds of the invention and related compounds for URAT1 inhibition were determined according to a similar method described in the literature (US 20140005136).

Construction of cell lines stably expressing the humanized URAT1 transporter: the humanized URAT1 gene (SLC 22A 112) was subcloned from plasmid pCMV6-XL-5 (origin) onto eukaryotic expression plasmid pCMV6/neo (origin). Gene sequencing enabled the humanization of URAT1 consistent with the information recorded in the gene library (NM-144585.2). HEK293 human embryonic kidney cells (ATC)C# CRL-1573) in EMEM tissue culture medium in 5% CO ₂ And culturing in an air atmosphere of 95%. pCMV6/Neo/URAT1 was transfected onto HEK293 cells using an L2000 type transfection reagent (invitrogen). After 24 hours, the transfected cells were divided into tissue culture dishes of 10cm diameter and grown for one day, after which the medium was replaced with fresh medium containing 0.5mg/ml G418 (Gibco). After 8 days, drug resistant colonies were selected and collected and tested for their transport activity on 14C-labeled uric acid. HEK293/URAT1 cells were seeded at a density of 75000 per well on poly-D-lysine coated 96-well plates.

These cells were grown overnight at 37℃in an incubator and then cooled to room temperature, wherein the culture broth was washed once with 250. Mu.L/well of wash solution (125 mM sodium gluconate, 10m MHEPES at pH=7.3). The test compound or blank was added to a buffer containing 40. Mu.M 14C-labeled uric acid (54 mCi/mmol) containing 125mM sodium gluconate, 4.8mM potassium gluconate, 1.2mM potassium dihydrogen phosphate, 1.2mM magnesium sulfate, 1.3mM calcium gluconate, 5.6mM glucose, 25mM HEPES, and a final pH=7.3. The 96-well plate was incubated at room temperature for 10 minutes, followed by three washes with 50. Mu.L/well and 250. Mu.L/well of each of the above-described washes. Microscint type 20 liquid flashing agent was added to 96 well plates and plates were incubated overnight at 45 ℃ and then read on TopCount Plate Reader and IC50 calculated therefrom. The results are shown in the following table.

IC50 value of example Compounds for URAT1

Compounds of formula (I)	IC50(nM)
		1	3.9
2	2.1
		3	3.0
4	1.1

The determination result of the IC50 shows that the compound provided by the invention has unexpected URAT1 inhibition effect, high activity and good selectivity, and can be used as a medicament for treating hyperuricemia and gout.

Claims (4)

1. A compound as shown below and pharmaceutically acceptable salts thereof:

2. a pharmaceutical composition comprising a compound of any one of claim 1 and pharmaceutically acceptable salts thereof and one or more pharmaceutically acceptable carriers.

3. Use of a compound as claimed in claim 1 and a pharmaceutically acceptable salt thereof in the manufacture of a medicament for inhibiting URAT 1.

4. The use according to claim 3, wherein the use is in the manufacture of a medicament for the treatment or prevention of a disorder associated with abnormal uric acid levels.