Aroyl nuciferine derivative inhibiting URAT1 and its preparation method and application

Abstract

The present invention belongs to the field of drug synthesis, and in particular relates to a URAT1-inhibiting aromatic acyl nuciferine derivative and its preparation method and application. The present invention structurally modifies nuciferine by introducing an active group, thereby improving the biological activity of nuciferine in lowering uric acid. Cell experiments have found that the aromatic acyl nuciferine derivative prepared by the present invention can inhibit the expression of URAT1 in HK-2 cells, promote uric acid excretion, thereby reducing uric acid levels, which is of great significance for the further development of new URAT1 inhibitors. The aromatic acyl nuciferine derivative with uric acid-lowering and kidney-protecting effects provided by the present invention has a simple synthesis method, low raw material price, and novel structure.

Description

Technical Field

[0001] The present invention belongs to the field of drug synthesis, and particularly relates to an aromatic acyl nuciferine derivative capable of inhibiting URAT1, and a preparation method and application thereof. Background Art

[0002] URAT1 is a member of the OATs (organic anion transporters) family, and its encoding gene is SLC22A12 URAT1 is a 12-transmembrane domain protein containing 555 amino acids. It is mainly expressed on the apical brush border membrane of the proximal tubular epithelial cells of the kidney. The concentration gradient and chemical gradient difference on both sides of the tubular lumen are used to transport urate from the proximal renal tubule lumen to the renal tubular epithelial cells, which is the first step in urate reabsorption. Studies have shown that URAT1 is only expressed in renal tissue and is the most important transporter involved in urate reabsorption. URAT1 has been identified as a high-affinity uric acid transporter and the main protein involved in uric acid reabsorption (about 90%), playing a dominant role in the transport of uric acid. Therefore, URAT1 has become an important target for uric acid-lowering drugs in recent years. In addition, single nucleotide polymorphisms (SNPs) in URAT1 have been shown to be associated with the pathogenesis of metabolic syndrome, including obesity, low high-density lipoprotein cholesterol, increased waist circumference, and essential hypertension.

[0003] Currently, clinical drugs for treating hyperuricemia include allopurinol, probenecid, benzbromarone, etc. These drugs can cause chronic damage to the urinary system, including the kidneys, and are prone to forming urate crystals and stones.

[0004] Nuciferine is a kind of Nelumbo nucifera Gaertn ) Alkaloids extracted from the plant. Pharmacological studies have shown that it possesses a variety of pharmacological activities, including antioxidant, anti-obesity, anti-inflammatory, anti-atherosclerotic, antiviral, and anti-tumor properties. Due to its low toxicity and minimal side effects, nuciferine has become a hot topic in pharmaceutical research and holds broad development prospects.

[0005] Wang Mingxing et al. discovered through experiments that nelumbo nucifera in lotus leaves can reduce serum urate and improve renal function. It can also inhibit the production of renal interleukins in mice with hyperuricemia caused by potassium oxonate. For the first time, it was reported that nelumbo nucifera can exert anti-inflammatory and anti-hyperuricemic effects by regulating organic ion transporters and inflammatory signals in renal hyperuricemia. The study found that daily use of supplements rich in nelumbo nucifera can effectively protect and prevent nephritic hyperuricemia. However, the uric acid-lowering activity of nelumbo nucifera is low and cannot be directly used as a clinical uric acid-lowering drug. Therefore, regulating the proteins involved in uric acid reabsorption or secretion is an effective strategy for preventing and treating hyperuricemia. Summary of the Invention

[0006] In view of the problems existing in the prior art, the present invention provides an aromatic acyl nuciferine derivative that inhibits URAT1.

[0007] The present invention also provides a preparation method of the aromatic acylnuciferine derivative.

[0008] The present invention further provides the use of the above-mentioned aromatic acyloylnuciferine derivatives in preparing URAT1 inhibitors.

[0009] The technical solutions adopted by the present invention to achieve the above-mentioned purpose are as follows:

[0010] The present invention provides an aromatic acyl nuciferine derivative that inhibits URAT1, and its molecular structure is as follows:

[0011] ;

[0012] R is 、 、 、 、 .

[0013] The present invention also provides a method for preparing the above-mentioned aromatic acyloylnuciferine derivative that inhibits URAT1, comprising the following steps:

[0014] (1) Dissolve nuciferine in methanol, then add hydrogen peroxide and stir to react; after the reaction is completed, adjust the pH value of the reaction solution, then add distilled water, extract with chloroform, dry the chloroform layer with anhydrous Na2SO4, filter to remove the solid, concentrate the filtrate under reduced pressure, separate and elute with silica gel column chromatography to obtain nuciferine- N - oxides;

[0015] (2) Under ice bath conditions, N The oxide is dissolved in methanol, and then a methanol solution of FeSO4·7H2O is added. The reaction is stirred after removing the ice bath to obtain a reaction solution. A saturated NaHCO3 solution is added to the reaction solution and the reaction is continued. After the reaction is completed, distilled water is added, and the mixture is extracted with chloroform. The chloroform layer is dried over anhydrous Na2SO4, the solid is removed by filtration, and the mixture is evaporated to dryness under reduced pressure. The mixture is separated by silica gel column chromatography and eluted to obtain protonuciferine.

[0016] (3) Dissolve proto-nuciferine and triethylamine in anhydrous DCM, slowly add RX dropwise, and stir to react; after the reaction is completed, quench the reaction, extract the reaction solution with dichloromethane, dry it with anhydrous sodium sulfate, concentrate the filtrate, and obtain the target compound by column chromatography.

[0017] Furthermore, in step (1), the ratio of nuciferine to hydrogen peroxide is 1 mmol:7.5 mL.

[0018] Furthermore, in step (1), the stirring reaction is carried out at room temperature for 24 hours; the pH is adjusted to 10 using a 3M NaOH aqueous solution; and elution is performed using an eluent consisting of chloroform: methanol: triethylamine in a volume ratio of 30:1:1‰.

[0019] Furthermore, in step (2), nuciferine- N -oxide and FeSO4·7H2O molar ratio is 1:2; the concentration of FeSO4·7H2O methanol solution is 1 mol / L; N -oxide and saturated NaHCO3 solution ratio is 1mmol:10mL;.

[0020] Furthermore, in step (2), the stirring reaction is carried out at room temperature for 12 hours; the reaction is continued for 20 minutes; and the eluent composed of chloroform: methanol: triethylamine in a volume ratio of 10:1:1‰ is used for elution.

[0021] Furthermore, in step (3), the molar ratio of protonuciferine, triethylamine and RX is 1:2-5:1-2.

[0022] Furthermore, in step (3), the temperature is maintained at 0°C during the dropwise addition of RX; the stirring reaction is stirred at room temperature for 10-12 hours; and saturated sodium bicarbonate is used to quench the reaction.

[0023] The present invention also provides the use of the above-mentioned aromatic acyl nuciferine derivatives in the preparation of URAT1 inhibitors.

[0024] The reaction route for preparing the aromatic acyl nuciferine derivatives of the present invention is:

[0025]

[0026] The beneficial effects of the present invention are:

[0027] (1) The present invention improves the biological activity of nuciferine in lowering uric acid by introducing active groups to modify its structure. Cell experiments have shown that the aromatic acyl nuciferine derivatives prepared by the present invention can inhibit the expression of URAT1 in HK-2 cells and promote uric acid excretion, thereby reducing uric acid levels. This is of great significance for the further development of new URAT1 inhibitors.

[0028] (2) The aromatic acyl nuciferine derivatives provided by the present invention have uric acid-lowering and kidney-protecting effects, and have a simple synthesis method, low raw material price, and novel structure. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 1H spectrum of compound 6 prepared in Example 1;

[0030] Figure 2 C chromatogram of compound 6 prepared in Example 1;

[0031] Figure 3 HRMS pattern of compound 6 prepared in Example 1;

[0032] Figure 4 Proton spectrum of compound 7 prepared in Example 2;

[0033] Figure 5 C chromatogram of compound 7 prepared in Example 2;

[0034] Figure 6 HRMS pattern of compound 7 prepared in Example 2;

[0035] Figure 7 1H spectrum of compound 26 prepared in Example 3;

[0036] Figure 8 C chromatogram of compound 26 prepared in Example 3;

[0037] Figure 9 HRMS pattern of compound 26 prepared in Example 3;

[0038] Figure 10 1H spectrum of compound 15 prepared in Example 4;

[0039] Figure 11 C12 spectra of compound 15 prepared in Example 4;

[0040] Figure 12 HRMS pattern of compound 15 prepared in Example 4;

[0041] Figure 13 1H spectrum of compound 18 prepared in Example 5;

[0042] Figure 14 C chromatogram of compound 18 prepared in Example 5;

[0043] Figure 15 HRMS spectrum of compound 18 prepared in Example 5. DETAILED DESCRIPTION

[0044] The technical solution of the present invention is further explained and illustrated by means of specific embodiments below.

[0045] Unless otherwise specified, the raw materials used in the present invention are commercially available.

[0046] Example 1

[0047] (1) Nuciferine (0.295 g, 1 mmol) was dissolved in 15 mL of methanol, and then 7.5 mL of 30% H2O2 was added to the solution. The mixture was stirred at room temperature for 24 h. After the reaction, 3M NaOH aqueous solution was added to the reaction solution to adjust the pH to 10. Then 150 mL of distilled water was added and the mixture was extracted with chloroform three times (600 mL×3). The chloroform layer was dried over anhydrous Na2SO4, the solid was removed by filtration, and the filtrate was concentrated under reduced pressure. The mixture was separated by silica gel column chromatography and eluted with chloroform: methanol: triethylamine (30:1:1‰ v / v / v) to obtain nuciferine- N -Oxide.

[0048] (2) Under ice bath conditions, N -oxide (0.310 g, 1 mmol) was dissolved in 2 mL of methanol, followed by the addition of a 2 mL solution of FeSO₄·7H₂O (0.304 g, 2 mmol) in methanol. The ice bath was removed and the reaction was stirred at room temperature for 12 h. Finally, 10 mL of saturated NaHCO₃ solution was added to the reaction solution and the reaction was continued for 20 min. After the reaction, 90 mL of distilled water was added to the three-necked flask, and the mixture was extracted three times with chloroform (100 mL x 3). The chloroform layer was dried over anhydrous Na₂SO₄, the solid was removed by filtration, and the mixture was evaporated to dryness under reduced pressure. Protonuciferine was obtained by silica gel column chromatography, eluting with chloroform:methanol:triethylamine (10:1:1‰).

[0049] (3) Proto-nuciferine (0.10 g, 0.31 mmol) and triethylamine (0.12 mL, 0.86 mmol) were dissolved in anhydrous DCM (20 mL). 3,5-Dichloro-4-methoxybenzoyl chloride (0.09 g, 0.37 mmol) was slowly added dropwise at 0°C. The reaction solution was stirred at room temperature overnight. After the reaction was completed, the reaction was quenched with saturated sodium bicarbonate (25 mL). The reaction solution was extracted with dichloromethane (2×25 mL) and dried over anhydrous sodium sulfate. The filtrate was concentrated and purified by column chromatography to obtain the target compound 6 as a white powder with mp 185.4–186.4 °C.

[0050] The structural formula of compound 6 is:

[0051]

[0052] like Figure 1-Figure 3 As shown:

[0053] 1 H NMR (600 MHz, CDCl3) δ 8.45 (d, J= 7.9 Hz, 1H), 7.40 (s, 2H), 7.36 –7.31 (m, 1H), 7.31 – 7.24 (m, 2H), 6.68 (s, 1H), 5.12 (s, 1H), 4.20 – 3.93(m, 4H), 3.91 (s, 3H), 3.68 (s, 3H), 3.29 (s, 1H), 3.12 (s, 1H), 3.01 – 2.85(m, 2H), 2.68 (d, J = 15.4 Hz, 1H).

[0054] 13 C NMR (151 MHz, CDCl3) δ 167.71, 153.41, 152.34, 145.98, 136.23,133.89, 131.48, 129.89, 128.72, 128.51, 127.92, 127.82, 127.60, 127.42,127.22, 125.70, 111.39.60.88, 60.07, 56.01, 50.40, 42.19, 34.54, 30.63.

[0055] HRMS calcd for C 26 H 23 Cl2NO4: [M+Na] + 506.0902, found 506.0902.

[0056] Example 2

[0057] Steps (1) and (2) are the same as in Example 1;

[0058] (3) Protothecin (0.10 g, 0.31 mmol) and triethylamine (0.12 mL, 0.86 mmol) were dissolved in anhydrous DCM (20 mL). Methyl p-chloroformylbenzoate (0.07 g, 0.35 mmol) was slowly added dropwise at 0°C. The reaction mixture was stirred at room temperature overnight. After the reaction was completed, the reaction was quenched with saturated sodium bicarbonate (25 mL). The reaction mixture was extracted with dichloromethane (2 × 25 mL) and dried over anhydrous sodium sulfate. The filtrate was concentrated and purified by column chromatography to obtain the target compound 7 as a white powder with mp 152.7–153.7 °C.

[0059] The structural formula of compound 7 is:

[0060]

[0061] like Figure 4-Figure 6 As shown:

[0062] 1 H NMR (600 MHz, CDCl3) δ 8.37 (d, J = 5.9 Hz, 1H), 8.04 (d, J = 5.1 Hz,2H), 7.44 (d, J = 7.6 Hz, 2H), 7.38 – 7.20 (m, 3H), 6.60 (s, 1H), 5.18 (s, 1H), 3.87 (s, 3H), 3.83 (s, 3H), 3.61 (s, 3H), 3.30 – 3.05 (m, 2H), 3.00 – 2.65(m, 3H), 2.57(d, J = 14.2 Hz, 1H).

[0063] 13 C NMR (151 MHz, CDCl3) δ 171.17, 169.72, 166.39, 152.29, 145.93,141.05, 136.37, 131.49, 131.05, 129.96, 128.78, 128.48, 127.89, 127.83,127.15, 126.67, 125.86, 111.40, 60.05, 56.00, 52.36, 50.77, 43.08, 34.34,30.66.

[0064] HRMS calcd for C 27 H 25 NO5: [M+Na] + 466.1630, found 466.1635.

[0065] Example 3

[0066] Steps (1) and (2) are the same as in Example 1;

[0067] (3) Proto-nuciferine (0.10 g, 0.31 mmol) and triethylamine (0.12 mL, 0.86 mmol) were dissolved in anhydrous DCM (20 mL). 4-Cyanobenzoyl chloride (0.06 g, 0.36 mmol) was slowly added dropwise at 0°C. The reaction mixture was stirred at room temperature overnight. After the reaction was completed, the reaction was quenched with saturated sodium bicarbonate (25 mL). The reaction mixture was extracted with dichloromethane (2 × 25 mL) and dried over anhydrous sodium sulfate. The filtrate was concentrated and purified by column chromatography to obtain the target compound 26 as a white powder with mp 215.4–216.4°C.

[0068] The structural formula of compound 26 is:

[0069]

[0070] like Figure 7-Figure 9 As shown:

[0071] 1 H NMR (600 MHz, CDCl3) δ 8.37 (d, J = 7.1 Hz, 1H), 7.68 (d, J = 5.6 Hz,2H), 7.49 (d, J = 7.6 Hz, 2H), 7.26 – 7.22 (m, 3H), 6.60 (s, 1H), 5.16 (s, 1H), 3.84 (s, 3H), 3.81 – 3.65 (m, 1H), 3.61 (s, 3H), 3.23 – 3.13 (m, 2H), 2.86 –2.84 (m, 2H), 2.61 – 2.58 (m, 1H).

[0072] 13 C NMR (151 MHz, CDCl3) δ 168.71, 152.38, 146.01, 141.06, 136.17,132.75, 132.57, 131.45, 128.52, 127.95, 127.82, 127.45, 127.25, 125.58,118.15, 113.42, 111.41, 60.07, 56.02, 50.93, 43.18, 34.30, 30.62.

[0073] HRMS calcd for C 26 H 22 N2O3: [M+Na] +433.1528, found 433.1524.

[0074] Example 4

[0075] Steps (1) and (2) are the same as in Example 1;

[0076] (3) Protothecin (0.10 g, 0.31 mmol) and triethylamine (0.12 mL, 0.86 mmol) were dissolved in anhydrous DCM (20 mL). 2-Thiophenecarbonyl chloride (0.06 g, 0.41 mmol) was slowly added dropwise at 0 °C. The reaction mixture was stirred at room temperature overnight. After the reaction was completed, the reaction was quenched with saturated sodium bicarbonate (25 mL). The reaction mixture was extracted with dichloromethane (2 × 25 mL) and dried over anhydrous sodium sulfate. The filtrate was concentrated and purified by column chromatography to obtain the target compound 15 as a white powder with mp 153.9–154.9 °C.

[0077] The structural formula of compound 15 is:

[0078]

[0079] like Figure 10-12 As shown:

[0080] 1 H NMR (600 MHz, DMSO) δ 8.30 (d, J = 7.9 Hz, 1H), 7.78 (dd, J = 5.0, 0.8Hz, 1H), 7.52 – 7.46 (m, 1H), 7.38 – 7.32 (m, 2H), 7.31 – 7.25 (m, 1H), 7.15(dd, J = 4.8, 3.8 Hz, 1H), 6.93 (s, 1H), 4.92 (dd, J = 13.6, 3.7 Hz, 1H), 4.43(d, J = 11.7 Hz, 1H), 3.84 (s, 3H), 3.61 (s, 3H), 3.34 (dd, J = 15.2, 9.3 Hz,1H), 3.02 (dd, J = 13.7, 4.0 Hz, 1H), 2.98 – 2.83 (m, 2H), 2.78 (d, J = 15.6 Hz,1H).

[0081] 13C NMR (151 MHz, CDCl3) δ 162.81, 151.20, 144.80, 136.91, 135.38,130.57, 128.17, 127.71, 127.54, 127.48, 127.41, 126.83, 126.69, 126.13,125.79, 125.07, 110.37, 59.01, 54.96, 51.44, 40.90, 34.06, 29.50.

[0082] HRMS calcd for C 23 H 21 NO3S: [M+Na] + 414.1140, found 414.1143.

[0083] Example 5

[0084] Steps (1) and (2) are the same as in Example 1;

[0085] (3) Protonuciferine (0.10 g, 0.31 mmol) and triethylamine (0.12 mL, 0.86 mmol) were dissolved in anhydrous DCM (20 mL). Cyclohexanecarbonyl chloride (0.06 g, 0.41 mmol) was slowly added dropwise at 0 °C. The reaction mixture was stirred at room temperature overnight. After the reaction was completed, the reaction was quenched with saturated sodium bicarbonate (25 mL). The reaction mixture was extracted with dichloromethane (2 × 25 mL) and dried over anhydrous sodium sulfate. The filtrate was concentrated and purified by column chromatography to obtain the target compound 18 as a light yellow powder with mp 144.0–145.0 °C.

[0086] The structural formula of compound 18 is:

[0087]

[0088] like Figure 13-15 As shown:

[0089] 1 H NMR (600 MHz, DMSO) δ 8.30 (d, J = 7.9 Hz, 1H), 7.85 (s, 1H), 7.38 –7.31 (m, 2H), 7.30 – 7.25 (m, 1H), 7.08 – 7.01 (m, 1H), 6.92 (s, 1H), 6.64(dd, J = 3.3, 1.7 Hz, 1H), 4.92 (dd, J= 13.2, 4.4 Hz, 1H), 4.55 (d, J = 11.8 Hz,1H), 3.84 (s, 3H), 3.61 (s, 3H), 3.30 (s, 1H), 3.07 – 2.83 (m, 3H), 2.79 (d, J = 15.5 Hz, 1H).

[0090] 13 C NMR (151 MHz, CDCl3) δ 158.62, 151.15, 147.25, 144.74, 142.78,135.66, 130.61, 128.43, 127.48, 127.30, 126.80, 126.66, 126.09, 125.09,115.39, 110.39, 110.31, 59.01, 54.95, 51.39, 39.44, 34.39, 29.37.

[0091] HRMS calcd for C 23 H 21 NO4: [M+Na] + 398.1368, found 398.1373.

[0092] Effect verification

[0093] (I) Cell culture and drug administration

[0094] Cell passaging. When HK-2 cells grow to a density of 85%-90%, the cells are passaged. Place the cells in a clean bench, discard the old culture medium with a pipette, add 4.0 mL of PBS to wash, discard the PBS, and repeat 2-3 times. Use a pipette to add 1.0 mL of 0.25% trypsin to digest for 40 seconds, aspirate the trypsin with a pipette, add culture medium and pipette repeatedly until it becomes a single cell suspension. Take 1.0 mL of cell suspension and transfer it to a new culture dish, add 7 mL of culture medium, shake well, and transfer the culture dish to a cell culture incubator after observation under a microscope.

[0095] Uric acid induction and drug administration. When HK-2 cells have successfully recovered and stably passaged for more than three generations, and the cell density in the culture dish reaches 85%-90%, remove the cells from the incubator and place them on a clean bench. Discard the culture medium and wash three times with PBS to remove dead cells. Add culture medium containing 100 mg / L uric acid and a certain amount of the compound solution prepared in the examples (final concentration of the compound is 50 μM) to the cell culture dishes in the model and drug groups, shake well, label, and transfer to an incubator for 48 hours for protein extraction and quantitative analysis.

[0096] (II) Cell protein quantification experiment

[0097] Take the protein standard solution from the BCA kit and add it to the protein standard to prepare a 1 mg / mL protein solution. Dilute the solution to concentrations of 0.5, 0.4, 0.2, 0.1, and 0.05 mg / mL for subsequent sample loading. Prepare the BCA colorimetric reagent in a 50.0 mL centrifuge tube according to the kit instructions. Add 25 µL of the diluted protein solution to a 96-well plate, setting up three replicates for each concentration. Thaw the extracted cell protein in advance and dilute it 40- to 80-fold. Add 25 µL of the protein solution to each well of a 96-well plate, setting up three replicates for each concentration. After protein loading, add 125 µL of the BCA colorimetric reagent and incubate in a constant temperature incubator for half an hour. Read at 562 nm using a microplate reader. Construct a protein standard curve based on the absorbance values. Substitute the absorbance values ​​of the protein to be tested into the curve to determine the protein concentration.

[0098] (3) Western-Blot

[0099] Prepare gel. Take a professional gel-making glass plate, clean it, and dry it. Clamp it onto a mold and inject the lower layer of gel prepared according to the instructions for preparing polyacrylamide gel (PAGE). Seal and flatten the gel surface with Wahaha water. Let it stand at room temperature for 20-30 minutes. Once the gel and water layers are clearly separated, discard the Wahaha water and inject the 5% upper layer of gel prepared according to the instructions. Insert a 10-well comb and let it stand at room temperature again for 20-30 minutes. Insert the prepared gel into the electrophoresis tank, fill it with the prepared 1x electrophoresis buffer, and remove the comb vertically. Add 5 µL of protein marker to each well and 60 µg of the quantified protein to the experimental well. Connect the power supply to a constant voltage of 120 V and set the desired time for electrophoresis. After electrophoresis, transfer the gel to the membrane. Cut a 10-well strip of polyvinylidene fluoride (PVDF) membrane and activate it in methanol before transferring the gel to the membrane. After electrophoresis, the constant current was 300 mA and electrotransfer was performed for 60-75 min.

[0100] The transferred PVDF membrane was cut along a protein marker to identify the target bands URAT1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The membrane was then placed in 3.0 mL of pre-chilled 5% milk blocking buffer and blocked on a shaker for 2 hours. The blocked target bands were placed in an antibody incubation box, and URAT1 (GLUT9) and GAPDH antibodies diluted according to the manufacturer's instructions were added. The membrane was incubated overnight at 4°C. The next day, the URAT1 (GLUT9) and GAPDH solutions were discarded, and the membrane was washed three times with TBST on a shaker for 5 minutes each. The corresponding secondary antibody was added and incubated on a shaker for another 2 hours. After binding to the secondary antibody, the membrane was washed, enhanced chemiluminescence reagent was added, and the membrane was scanned and photographed using a chemiluminescence imaging system. The grayscale values ​​of the protein bands were analyzed using ImageJ, and GraphPad Prism 8.0 software was used to plot the expression of URAT1 (GLUT9) in each cell group. The percentage of protein expression was calculated based on the URAT1 protein expression in the blank group. The results are shown in Table 1.

[0101] Table 1 Experimental results of compounds (50 μM) inhibiting URAT1 expression in HK-2 cells

[0102]

Claims

1. An aromatic acyl nuciferine derivative that inhibits URAT1, characterized in that Its molecular structure is: ； R is 、 、 、 、 .

2. A method for preparing an aromatic acyl nuciferine derivative that inhibits URAT1 according to claim 1, characterized in that: The following steps are involved: (1) Dissolve nuciferine in methanol, then add hydrogen peroxide and stir to react; after the reaction is completed, adjust the pH value of the reaction solution, then add distilled water, extract with chloroform, dry the chloroform layer with anhydrous Na2SO4, filter to remove the solid, concentrate the filtrate under reduced pressure, separate and elute with silica gel column chromatography to obtain nuciferine- N - oxides; (2) Under ice bath conditions, N The oxide is dissolved in methanol, and then a methanol solution of FeSO4·7H2O is added. The reaction is stirred after removing the ice bath to obtain a reaction solution. A saturated NaHCO3 solution is added to the reaction solution and the reaction is continued. After the reaction is completed, distilled water is added, and the mixture is extracted with chloroform. The chloroform layer is dried over anhydrous Na2SO4, the solid is removed by filtration, and the mixture is evaporated to dryness under reduced pressure. The mixture is separated by silica gel column chromatography and eluted to obtain protonuciferine. (3) Dissolve proto-nuciferine and triethylamine in anhydrous DCM, slowly add acyl chloride dropwise, and stir to react; after the reaction is completed, quench the reaction, extract the reaction solution with dichloromethane, dry it with anhydrous sodium sulfate, concentrate the filtrate, and obtain the target compound by column chromatography.

3. The preparation method according to claim 2, characterized in that In step (1), the ratio of nuciferine to hydrogen peroxide is 1 mmol:7.5 mL.

4. The preparation method according to claim 2 or 3, characterized in that In step (1), the stirring reaction is carried out at room temperature for 24 hours; the pH is adjusted to 10 using a 3M NaOH aqueous solution; and elution is performed using an eluent consisting of chloroform: methanol: triethylamine in a volume ratio of 30:1:1‰.

5. The preparation method according to claim 2, characterized in that In step (2), nuciferine- N -oxide and FeSO4·7H2O molar ratio is 1:2; the concentration of FeSO4·7H2O methanol solution is 1 mol / L; N The ratio of -oxide and saturated NaHCO3 solution is 1 mmol:10 mL.

6. The preparation method according to claim 2 or 5, characterized in that In step (2), the stirring reaction is carried out at room temperature for 12 hours; the reaction is continued for 20 minutes; and the eluent composed of chloroform: methanol: triethylamine in a volume ratio of 10:1:1‰ is used for elution.

7. The preparation method according to claim 2, characterized in that In step (3), the molar ratio of protonuciferine, triethylamine and acyl chloride is 1:2-5:1-2.

8. The preparation method according to claim 2 or 7, characterized in that In step (3), the temperature is maintained at 0°C during the dropwise addition of the acyl chloride; the stirring reaction is carried out at room temperature for 10-12 hours; and saturated sodium bicarbonate is used to quench the reaction.

9. Use of the aromatic acyloylnuciferine derivative according to claim 1 in the preparation of a URAT1 inhibitor.

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