Methylated aromatic substituted genipin derivative with uric acid reducing effect as well as preparation method and application of methylated aromatic substituted genipin derivative

Abstract

The invention belongs to the field of preparation of functional compounds, and particularly relates to a methylated aromatic substituted genipin derivative with a uric acid reducing effect as well as a preparation method and application of the methylated aromatic substituted genipin derivative. The genipin derivative synthesized by the invention is subjected to XOD inhibitory activity screening, and researches on in-vivo and in-vitro activity, uric acid reduction and XOD inhibitory activity, anti-inflammatory activity and anti-fibrosis activity show that the derivative shows relatively high XOD inhibitory activity; oxidative stress is relieved by inhibiting XOD activity at the liver, so that liver inflammation is inhibited. The invention provides a structural and theoretical basis for subsequent further research on a more efficient XOD inhibitor.

Description

Technical Field

[0001] This invention belongs to the field of functional compound preparation, specifically relating to a methylated aromatic substituted genipin derivative with uric acid-lowering effect, its preparation method, and its application. Background Technology

[0002] Hyperuricemia is a metabolic disease caused by disordered metabolism of purines in the body. In recent years, the number of patients with hyperuricemia has been rising, making it the second most common metabolic disease. Long-term hyperuricemia can lead to many complications, especially kidney inflammation, and in severe cases, renal fibrosis, seriously damaging kidney function. Xanthine oxidase (XOD) plays an important role in uric acid metabolism and is a key enzyme regulating uric acid production. Therefore, most current uric acid-lowering drugs target XOD, such as allopurinol and febuxostat. However, most of these XOD inhibitors have significant toxic side effects, causing liver and kidney damage. Therefore, developing novel XOD inhibitors with high XOD inhibitory activity while also promoting kidney repair has become an urgent problem to be solved. Summary of the Invention

[0003] To address the problems existing in the prior art, the present invention provides a methylated aromatic substituted genipin derivative with uric acid-lowering effect.

[0004] Another object of the present invention is to provide a method for preparing the above-mentioned methylated aromatic substituted genipin derivative.

[0005] This invention also provides the application of methylated aromatic substituted genipin derivatives in the preparation of uric acid-lowering drugs.

[0006] The technical solution adopted by the present invention to achieve the above objectives is as follows: This invention provides a methylated aromatic substituted genipin derivative with uric acid-lowering activity, wherein the structural formula of the methylated aromatic substituted genipin derivative is as follows: ; Where R1 is , , , , , , .

[0007] Another object of the present invention is to provide a method for preparing the above-mentioned methylated aromatic substituted genipin glycoside derivative, comprising the following steps: (1) Add genipin and triphenylphosphine to a round-bottom flask, and slowly add carbon tetrabromide while stirring; after the addition is complete, react at room temperature; after washing the organic phase, concentrate the organic phase and purify it by silica gel column chromatography to obtain compound 1; (2) At 0°C, an organic alcohol was added to the anhydrous dichloromethane stirring solution of compound 1 and stirred overnight at room temperature. After the reaction was completed, the solution was added to ice water and extracted with ethyl acetate. The organic phases were combined, washed, dried, concentrated, and purified by silica gel column chromatography to obtain compound 2a-2g, i.e., methylated aromatic substituted genipin glycoside derivative.

[0008] Preferably, in step (1), genipin and 0.1 equivalent of triphenylphosphine are mixed; the amount of carbon tetrabromide added is 1.3 equivalents; the washing is carried out sequentially using saturated sodium bisulfite aqueous solution, sodium bicarbonate aqueous solution and water; the eluent for silica gel column chromatography purification is petroleum ether: ethyl acetate = 10:1-1:1.

[0009] Preferably, in step (1), the reaction is carried out at room temperature for 8 hours.

[0010] Preferably, in step (1), the structural formula of compound 1 is: Preferably, in step (2), the concentration of compound 1 in anhydrous dichloromethane is 0.065-0.070 mol / mL; 1.2 equivalents of organic alcohol are added to the anhydrous dichloromethane solution of compound 1; the eluent for the silica gel column chromatography purification is methanol:dichloromethane = 30:1-5:1.

[0011] Preferably, in step (2), the organic alcohol is 3-hydroxyphenylacetylene, 4-hydroxyindole, m-trifluoromethylphenol, p-hydroxybenzonitrile, p-methoxyphenol, p-hydroxybenzaldehyde, or 1,3-methylenedioxy-4-phenol.

[0012] Preferably, in step (2), the reaction is carried out by stirring at room temperature overnight for 10-12 hours.

[0013] This invention also provides the application of the above-mentioned methylated aromatic substituted genipin glycoside derivatives in the preparation of uric acid-lowering drugs.

[0014] The present invention further provides the application of the above-mentioned methylated aromatic substituted genipin glycoside derivative as an XOD inhibitor with kidney repair function.

[0015] The reaction route diagram of this invention is as follows: This invention uses genipin as a lead compound, chemically modifies it, and designs and synthesizes genipin derivatives. To enhance the binding affinity of the genipin derivatives to XOD, the C-14 position of genipin is substituted, introducing an aromatic ring or heterocyclic group containing an organic alcohol. The chemical structure of the genipin derivatives is then analyzed. 1H NMR, 13 Characterization and confirmation were performed using C NMR.

[0016] The beneficial effects of this invention are as follows: The genipin derivative synthesized in this invention underwent XOD inhibitory activity screening. In vivo and in vitro studies on its activity, uric acid-lowering and XOD-inhibiting activities, anti-inflammatory activity, and anti-fibrotic activity showed that this derivative exhibited high XOD inhibitory activity; by inhibiting XOD activity in the liver, it reduces oxidative stress and thus inhibits liver inflammation. This invention provides a structural and theoretical basis for further research on more efficient XOD inhibitors. Attached Figure Description

[0017] Figure 1 Effects of genipin derivative 2d on serum related parameters in hyperuricemic mice. (ac) Effects of genipin derivative 2b on serum uric acid, blood urea nitrogen, and creatinine. (de) Effects of genipin derivative 2d on serum ALP and ALT levels in hyperuricemic mice; Figure 2 The effect of genipin derivatives 2d on renal pathological changes in hyperuricemic mice; Figure 3 The effect of genipin derivative 2d on renal fibrosis in hyperuricemic mice. Detailed Implementation

[0018] The technical solution of the present invention will be further explained and described below through specific embodiments.

[0019] The reagents and instruments used in this invention were as follows: Genipin and other chemicals were purchased from Aladdin (Shanghai, China), and routine reagents were purchased from Fuyu (Shanghai, China); thin-layer chromatography (TLC) plates and silica gel were purchased from Sinopharm (Shandong, China); nuclear magnetic resonance (NMR) spectra were recorded using a Bruker (Germany) NMR spectrometer in deuterated dimethyl sulfoxide (DMSO-d6) or deuterated chloroform (CDCl3).

[0020] Xanthine and XOD were purchased from Yuan Ye (Shanghai, China); allopurinol was purchased from Maclean's (Shanghai, China); commercial kits for AST, ALT, UA, Cr, and BUN were purchased from Jiancheng (Nanjing, China); and a commercial XOD kit was purchased from Beyotime (Shanghai, China).

[0021] Example 1: Compound 1 Genipin (100 mg, 0.442 mol) and 0.1 equivalent of triphenylphosphine (Ph3P) were added to a round-bottom flask, followed by the slow addition of 1.3 equivalents of carbon tetrabromide (CBr4) with stirring. After the addition was complete, the reaction was allowed to proceed at room temperature. The organic phase was washed successively with saturated aqueous solutions of sodium bisulfite, sodium bicarbonate, and water. The organic phase was concentrated and analyzed by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1–1:1) to give compound 1 (bromomethyl-genipin). The structural formula of compound 3 is as follows: Compound 1 (bromomethyl-genipin) is a yellow powder with a yield of 63.38% and a purity of 97.18%.

[0022] 1 H NMR (400 MHz, CDCl3) δ 7.27 (d, J = 13.9 Hz, 1H), 5.98 (d, J = 14.1 Hz,1H), 4.55-4.48 (m, 1H), 4.19-4.15 (m, 2H), 3.76-3.73 (m, 5H), 3.55 (d, J = 3.5Hz, 1H), 3.08 (dt, J = 42.7, 8.7 Hz, 1H), 2.93-2.89 (m, 1H), 2.82 (dd, J = 13.0, 4.9 Hz, 1H), 2.12 (dd, J = 17.1, 8.1 Hz, 1H). 13 C NMR (100 MHz, CDCl3) δ 167.67, 152.17, 139.62, 133.11, 110.91,102.21, 57.01, 51.25, 45.80, 39.07, 35.45, 30.26. Example 2: Compound 2a At 0 °C, 1.2 equivalents of 3-hydroxyphenylacetylene were added to a solution of compound 1 (100 mg, 0.330 mmol) in anhydrous dichloromethane (5 mL), and the mixture was stirred overnight at room temperature. After the reaction was complete, the solution was added to ice water (30 mL) and extracted with ethyl acetate. The organic phases were combined, washed with citric acid aqueous solution, dried, concentrated, and purified by silica gel column chromatography (methanol:dichloromethane = 30:1-5:1) to give compound 2a (m-ethynylphenyl-methylgenipin ether). The structural formula of compound 2a is: Compound 2a (m-ethynylphenyl-methyl genipin ether) is a transparent oily liquid with a yield of 50.37% and a purity of 95.29%.

[0023] 1 H NMR (400 MHz, CDCl3) δ 7.47 – 7.43 (m, 1H), 7.18 (d, J = 8.6 Hz, 1H),7.17 – 7.11 (m, 1H), 7.01 (d, J = 7.6 Hz, 1H), 6.85 (dt, J = 6.6, 3.3 Hz, 1H), 5.88 (d, J = 27.7 Hz, 1H), 4.61 (t, J = 12.0 Hz, 1H), 4.56 – 4.52 (m, 1H), 4.45(t, J = 6.6 Hz, 1H), 3.66 (s, 3H), 3.49 (d, J = 8.9 Hz, 3H), 2.99 (s, 1H), 2.83(dd, J = 16.6, 8.4 Hz, 1H), 2.61 (t, J = 7.6 Hz, 1H), 2.03 (tdd, J = 9.0, 6.3, 3.4Hz, 1H). 13 C NMR (100 MHz, CDCl3) δ 167.78, 158.36, 152.14, 138.62, 130.30,129.41, 124.80, 123.05, 117.80, 116.18, 110.98, 83.57, 66.40, 57.04, 51.25,46.16, 38.99, 35.53. Example 3: Compound 2b At 0 °C, 1.2 equivalents of 4-hydroxyindole were added to a solution of compound 3 (100 mg, 0.330 mmol) in anhydrous dichloromethane (5 mL), and the mixture was stirred overnight at room temperature. After the reaction was complete, the solution was added to ice water (30 mL) and extracted with ethyl acetate. The organic phases were combined, washed with citric acid aqueous solution, dried, concentrated, and purified by silica gel column chromatography (methanol:dichloromethane = 30:1-5:1) to give compound 2b (benzopyrrole-methylgenipin ether). The structural formula of compound 2b is: Compound 2b (benzopyrrole-methylgenipin ether) is a transparent oily liquid with a yield of 43.59% and a purity of 96.92%.

[0024] 1 H NMR (400 MHz, CDCl3) δ 7.44 (s, 1H), 6.97-6.91 (m, 2H), 6.74-6.66(m, 1H), 6.33 (dd, J = 12.8, 5.2 Hz, 2H), 5.41 (d, J = 15.1 Hz, 1H), 4.82-4.69(m, 2H), 3.63 (s, 3H), 3.59 (s, 3H), 2.73 (dd, J = 16.7, 8.9 Hz, 1H), 2.69-2.61(m, 1H), 2.35 (t, J = 7.6 Hz, 1H), 1.95-1.89 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 167.74, 152.14, 150.57, 139.99, 139.47,137.22, 129.70, 126.69, 122.71, 117.90, 110.91, 103.89, 103.21, 100.91,57.25, 51.25, 46.69, 45.88, 38.85, 36.07. Example 4: Compound 2c At 0 °C, m-trifluoromethylphenol (1.2 equivalents) was added to a solution of compound 1 (100 mg, 0.330 mmol) in anhydrous dichloromethane (5 mL), and the mixture was stirred overnight at room temperature. After the reaction was complete, the solution was added to ice water (30 mL) and extracted with ethyl acetate. The organic phases were combined, washed with citric acid aqueous solution, dried, concentrated, and purified by silica gel column chromatography (methanol:dichloromethane = 30:1-5:1) to give compound 2c (m-trifluorophenyl-methylgenipin ether). The structural formula of compound 2c is: Compound 2c (m-trifluorophenyl-methyl genipin ether) is a yellow oily liquid with a yield of 57.26% and a purity of 95.51%.

[0025] 1 H NMR (400 MHz, CDCl3) δ 7.46 (d, J = 0.6 Hz, 1H), 7.30 (t, J = 8.0 Hz, 1H), 7.12 (d, J = 7.7 Hz, 1H), 7.07 (s, 1H), 7.01 (dd, J = 8.3, 2.2 Hz, 1H), 5.90(d, J = 24.1 Hz, 1H), 4.66 (d, J = 13.3 Hz, 1H), 4.57 (dd, J = 16.7, 14.9 Hz, 2H),4.49-4.42 (m, 1H), 3.68 – 3.64 (m, 5H), 3.52-3.47 (m, 3H), 3.19-3.11 (m, 1H),2.84 (dd, J = 16.7, 8.4 Hz, 1H), 2.64-2.58 (m, 1H), 2.07-2.00 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 167.87, 167.75, 158.77, 152.15, 138.36,130.66, 129.94, 118.27, 117.50, 117.47, 111.48, 111.46, 111.44, 110.96,102.60, 99.78, 66.62, 57.01, 56.27, 51.25, 46.13, 38.99, 35.60. Example 5: Compound 2d At 0 °C, p-cyanophenol (1.2 equivalents) was added to a solution of compound 3 (100 mg, 0.330 mmol) in anhydrous dichloromethane (5 mL), and the mixture was stirred overnight at room temperature. After the reaction was complete, the solution was added to ice water (30 mL) and extracted with ethyl acetate. The organic phases were combined, washed with citric acid aqueous solution, dried, concentrated, and purified by silica gel column chromatography (methanol:dichloromethane = 30:1-5:1) to give compound 2d (p-cyanophenyl-methyl genipin ether). The structural formula of compound 2d is: Compound 2d (p-cyanophenyl-methyl genipin ether) is a yellow oily liquid with a yield of 53.84% and a purity of 96.13%.

[0026] 1 H NMR (400 MHz, CDCl3) δ 7.46 (d, J = 0.6 Hz, 1H), 7.19 (s, 1H), 7.14(t, J = 8.0 Hz, 1H), 7.01 (d, J = 7.6 Hz, 1H), 6.95 (d, J = 2.2 Hz, 1H), 5.86 (s,1H), 4.62 (d, J = 13.4 Hz, 1H), 4.56-4.52 (m, 1H), 4.46 (d, J = 7.8 Hz, 1H), 3.66(s, 2H), 3.50 (s, 2H), 2.99 (s, 1H), 2.83 (dd, J = 16.6, 8.4 Hz, 1H), 2.61 (t, J = 7.6 Hz, 1H), 2.06-2.01 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 167.78, 158.36, 152.14, 138.62, 130.30,129.41, 117.80, 116.18, 115.92, 110.98, 102.60, 66.40, 57.04, 51.25, 46.16,38.99, 35.53. Example 6: Compound 2e At 0 °C, p-methoxyphenol (1.2 equivalents) was added to a solution of compound 3 (100 mg, 0.330 mmol) in anhydrous dichloromethane (5 mL), and the mixture was stirred overnight at room temperature. After the reaction was complete, the solution was added to ice water (30 mL) and extracted with ethyl acetate. The organic phases were combined, washed with citric acid aqueous solution, dried, concentrated, and purified by silica gel column chromatography (methanol:dichloromethane = 30:1-5:1) to give compound 2e (p-methoxyphenyl-methylgenipin ether). The structural formula of compound 2e is: Compound 2e (p-methoxyphenyl-methyl genipin ether) is a yellow oily liquid with a yield of 66.39% and a purity of 97.04%.

[0027] 1 H NMR (400 MHz, CDCl3) δ 7.47-7.42 (m, 1H), 6.98 (t, J = 12.4 Hz, 2H), 6.76-6.71 (m, 2H), 5.91-5.82 (m, 1H), 4.49 (dd, J = 12.0, 4.7 Hz, 1H), 3.65 (s,3H), 3.47- 3.45 (m, 3H), 3.31-3.21 (m, 3H), 3.18-3.10 (m, 1H), 2.82 (dd, J =16.5, 8.4 Hz, 1H), 2.63 (t, J = 7.7 Hz, 1H), 2.03 (ddt, J = 15.1, 11.7, 4.7 Hz, 1H). 13 C NMR (100 MHz, CDCl3) δ 167.83, 156.56, 152.13, 150.62, 129.94,129.87, 114.60, 114.42, 102.56, 66.41, 57.01, 56.98, 56.33, 51.24, 46.24,38.97, 35.40, 20.48. Example 7: Compound 2f At 0 °C, p-hydroxybenzaldehyde (1.2 equivalents) was added to a solution of compound 3 (100 mg, 0.330 mmol) in anhydrous dichloromethane (5 mL), and the mixture was stirred overnight at room temperature. After the reaction was complete, the solution was added to ice water (30 mL) and extracted with ethyl acetate. The combined organic phases were washed with citric acid aqueous solution, dried, concentrated, and purified by silica gel column chromatography (methanol:dichloromethane = 30:1-5:1) to give compound 2f (p-aldehyde phenyl-methyl genipin ether). The structural formula of compound 2f is: Compound 2f (p-aldehyde phenyl-methyl genipin ether) is a yellow oily liquid with a yield of 52.37% and a purity of 95.14%.

[0028] 1 H NMR (400 MHz, CDCl3) δ 9.83-9.77 (m, 1H), 7.75 (t, J = 11.2 Hz, 2H),7.21-7.17 (m, 1H), 6.98-6.91 (m, 2H), 5.92 (d, J = 23.3 Hz, 1H), 4.74-4.61 (m,2H), 4.50-4.42 (m, 1H), 3.68-3.62 (m, 3s), 3.54-3.47 (m, 2H), 3.20-3.12 (m,1H), 2.85 (dd, J = 16.7, 8.5 Hz, 1H), 2.65-2.56 (m, 1H), 2.09-2.00 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 190.79, 167.71, 163.71, 152.14, 138.10,131.97, 130.94, 130.03, 115.00, 102.60, 66.66, 57.07, 46.15, 39.02, 35.63. Example 8: 2g of compound At 0 °C, 1,3-methylenedioxy-4-phenol (1.2 equivalents) was added to a solution of compound 3 (100 mg, 0.330 mmol) in anhydrous dichloromethane (5 mL), and the mixture was stirred overnight at room temperature. After the reaction was complete, the solution was added to ice water (30 mL) and extracted with ethyl acetate. The organic phases were combined, washed with citric acid aqueous solution, dried, concentrated, and purified by silica gel column chromatography (methanol:dichloromethane = 30:1-5:1) to give compound 2 g (1,3-methylenedioxyphenyl-methylgenipin ether). The structural formula of compound 2 g is: Compound 2g (1,3-methylenedioxyphenyl-methylgenipin ether) is a yellow oily liquid with a yield of 39.76% and a purity of 96.39%.

[0029] 1 H NMR (400 MHz, CDCl3) δ 7.44 (d, J = 11.0 Hz, 1H), 7.22-7.17 (m, 1H), 6.65 -6.58 (m, 1H), 6.45-6.42 (m, 1H), 6.29-6.23 (m, 1H), 5.85 (s, 1H), 4.54(d, J = 13.3 Hz, 1H), 4.46 (dd, J = 15.6, 7.8 Hz, 3H), 3.65 (s, 3H), 3.48 (s,3H), 3.16 (dd, J = 16.7, 7.8 Hz, 1H), 2.84-2.79 (m, 1H), 2.61 (t, J = 7.7 Hz,1H), 2.06-1.99 (m, 1H). 13 C NMR (100 MHz, CDCl3) δ 167.80, 154.18, 152.13, 148.20, 141.70,139.03, 130.15, 111.00, 107.92, 106.09, 102.56, 101.13, 98.25, 67.30, 56.99,51.24, 46.17, 38.96, 35.42. Example 1 (I) Determination of the in vitro XOD inhibitory activity of the compound The test compounds were diluted with phosphate-buffered saline (PBS) to different concentrations (200, 100, 50, 20, 10, 1, 0.5, and 0.25 μM). Simultaneously, blank and positive control groups were established by replacing the test compounds with PBS and allopurinol, respectively. The reaction was carried out in 96-well microplates, with 200 μL of reaction mixture per well. First, each concentration of test compound (100 μL) was mixed with XOD (50 U / L, 50 μL) and equilibrated at 37 °C for 5 minutes; then, xanthine (0.5 mM, 50 μL) was added to initiate the reaction, and the reaction system was incubated at 37 °C for 30 minutes. After the reaction was complete, the absorbance of each well at 295 nm was measured using a UV spectrophotometer.

[0030] Inhibition rate (%) = (Ab-At) / Ab×100%, where Ab is the absorbance of the blank control group and At is the absorbance of the test group.

[0031] The half-maximal inhibitory concentration (IC50) of genipin derivatives against xanthine oxidase (XOD) 50 As shown in Table 1.

[0032] Table 1 The results are shown in Table 1. Most compounds exhibited higher inhibitory activity than the parent compound genipin. Among them, compound 2d (IC50) showed the best inhibitory activity. 50 = 2.58µM) has the best activity.

[0033] (II) In vivo activity evaluation of the compound 2 days later (1) Establishment of a mouse model of hyperuricemia Male Kunming (KM) mice (20±2g) were purchased from Pengyue (Jinan, China). First, the mice were acclimatized to the laboratory environment for one week, during which time they were fed a standard diet. Then, all mice were randomly divided into two groups: a control group and an experimental group. Based on previous literature, a hyperuricemia model was established by administering adenine (AN) and potassium oxonate (PO).

[0034] Control group (NC): Mice were given 0.5% sodium carboxymethyl cellulose (CMC-Na) (10 mL / kg); Experimental group: Mice were given 0.5% CMC-Na (10 mL / kg), AN (200 mg / kg) and PO (500 mg / kg) for two weeks.

[0035] Two weeks later, uric acid levels in the control and experimental groups were measured to assess the successful establishment of the hyperuricemia mouse model. After successful model establishment, the experimental group mice were randomly divided into three groups, each receiving daily oral administration of the corresponding drug for two weeks: Model group: Mice were given 0.5% CMC-Na (10 mL / kg); Allopurinol: Mice were given 0.5% CMC-Na (10 mL / kg) and allopurinol (10 mg / kg); Compound 2d: Mice were given 0.5% CMC-Na (10 mL / kg) and Compound 2d (20 mg / kg).

[0036] (2) Mouse sample collection After drug administration, mice in each group were fasted for 12 hours, then anesthetized with chloroform hydrate to collect blood samples. Finally, the mice were euthanized, and the livers and kidneys were quickly collected. Blood samples were left at room temperature for 30 minutes, then centrifuged at 4°C and 12,000 rpm for 15 minutes to obtain serum, which was stored at -20°C for subsequent testing. The livers and kidneys were quickly separated and stored at -80°C for subsequent testing.

[0037] (3) Determination of UA, Cr, BUN, ALT and ALP Following the instructions of the commercial reagent kit, the levels of UA, Cr, BUN, ALT, and alkaline ALP were determined using the commercial kit. The absorbance of each indicator was obtained using a UV spectrophotometer, and the concentration was calculated according to the formulas in the kit instructions.

[0038] like Figure 1 As shown, compound 2d can effectively reduce serum-related indicators in hyperuricemic mice, indicating that compound 2d has significant liver and kidney protective effects.

[0039] (4) HE staining First, tissues fixed and preserved in 4% paraformaldehyde for 3 days were removed, dehydrated with a series of gradient concentrations of ethanol, then embedded in paraffin, and cut into 4 μm thick sections. The sections were baked at 65°C for 6 hours in an incubator, followed by hematoxylin-eosin staining. Pathological damage was observed under an optical microscope at 200x magnification, and detailed pathological analysis was performed.

[0040] like Figure 2 As shown, HE staining results indicated extensive renal tubular dilation in mice with hyperuricemia, vacuolation of renal tubular epithelial cells, unclear renal tubular structure, connective tissue hyperplasia, and punctate infiltration of neutrophils. A small number of renal tubular epithelial cells showed necrosis and nuclear fragmentation, and brush border detachment was frequently observed in the tubules. This indicated severe renal inflammation and damage in the hyperuricemia group. Furthermore, mice in the allopurinol group also exhibited significant vacuolation and other pathological changes in their kidneys. Compound 2d can reverse kidney damage and effectively improve renal pathological changes.

[0041] (5) Masson staining Masson staining was used to analyze tissue fibrosis. Using Image Pro-Plus software, the area of ​​collagen tissue was quantitatively measured by drawing lines around the positively stained areas, and then the average proportion of each microscopic field of view was calculated and plotted.

[0042] like Figure 3 As shown, Masson staining results revealed significant renal tubular dilation, focal inflammatory cell infiltration, and marked renal fibrosis in the interstitial region of mice with hyperuricemia. The allopurinol group showed significant interstitial fibrosis with numerous tissue vacuoles and multiple inflammatory manifestations in the kidneys. In contrast, the kidney tissue of mice treated with compound 2d showed uniform staining, normal glomerular morphology and structure, clear tubular structure, tightly packed tubular epithelial cells, and numerous focal lymphocyte infiltrations around local blood vessels. This indicates that Genipin 2d has a certain protective effect against renal fibrosis induced by hyperuricemia.

Claims

1. A methylation of aromatic-substituted genipin derivative having a uric acid-lowering effect, characterized by, The structural formula of the methylated aromatic-substituted genipin derivative is: ； wherein R1 is , , , , , , .

2. A process for preparing the methylation of the aromatic substituent of the geniposide derivative according to claim 1, characterized by, The method comprises the following steps: (1) adding genipin and triphenylphosphine into a round-bottom flask, slowly adding carbon tetrabromide under stirring, and reacting at room temperature after the addition; after washing the organic phase, the organic phase is concentrated, and compound 1 is obtained by silica gel column chromatography; (2) adding an organic alcohol to a stirring solution of compound 1 in anhydrous dichloromethane at 0 ℃, stirring at room temperature overnight; after the reaction is completed, the solution is added to ice water, extracted with ethyl acetate; the organic phases are combined, washed, and dried, and compound 2a-2g, i.e., the methylated aromatic-substituted genipin glycoside derivative, is obtained by concentration and silica gel column chromatography.

3. The production method according to claim 2, characterized by, In step (1), genipin is mixed with 0.1 equivalent of triphenylphosphine; the amount of carbon tetrabromide added is 1.3 equivalents; the washing is performed in sequence with saturated sodium bisulfite aqueous solution, sodium bicarbonate aqueous solution and water; and the eluent for silica gel column chromatography is petroleum ether: ethyl acetate = 10:1-1:

1.

4. The production method according to claim 2 or 3, characterized by, In step (1), the reaction is performed at room temperature for 8 hours.

5. The method of any one of claims 2-4, wherein, In step (1), the structural formula of compound 1 is: 。 6. The preparation method according to claim 2, characterized in that, In step (2), the concentration of compound 1 in anhydrous dichloromethane is 0.065-0.070 mol / mL; 1.2 equivalents of an organic alcohol are added to the anhydrous dichloromethane solution of compound 1; and the eluent for silica gel column chromatography is methanol:dichloromethane = 30:1-5:

1.

7. The production method according to claim 6, wherein In step (2), the organic alcohol is 3-hydroxyphenylacetylene, 4-hydroxyindole, m-trifluoromethylphenol, p-hydroxybenzonitrile, p-methoxyphenol, p-hydroxybenzaldehyde or 1,3-methylenedioxy-4-phenol.

8. The production method according to claim 2, 5 or 6, characterized by, In step (2), the reaction is stirring at room temperature overnight, and the reaction time is 10-12 hours.

9. Use of the methylated aromatic-substituted genipin glycoside derivative of claim 1 in the preparation of a drug for reducing uric acid.

10. Use of the methylated aromatic-substituted genipin glycoside derivative of claim 1 as an XOD inhibitor with a kidney repair effect.

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