A biphenyl triazole derivative, and a preparation method and application thereof

Abstract

This invention belongs to the field of medicinal chemistry, specifically relating to a biphenyltriazole derivative, its preparation method, and its application. The structural formula of this biphenyltriazole derivative is shown in Formula I. The biphenyltriazole-containing compound provided by this invention can inhibit hyaluronidase activity with mild cytotoxicity, and has the potential to be developed into a hyaluronidase inhibitor and applied in the production of drugs that inhibit hyaluronidase or products containing hyaluronic acid. The biphenyltriazole derivative significantly inhibits LPS-stimulated nitric oxide release from RAW264.7, indicating a certain anti-inflammatory activity.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology, specifically relating to a biphenyltriazole derivative, its preparation method, and its application. Background Technology

[0002] Hyaluronic acid (HA) is a polysaccharide composed of alternating D-glucuronic acid and N-acetyl-D-glucosamine. It plays a crucial role in the development and regulation of the human body, including fertilization, embryonic development, cell migration and differentiation, organ formation, wound healing, inflammation, and tumor growth and metastasis. Hyaluronidase is an enzyme that degrades hyaluronic acid and can be derived from eukaryotes, prokaryotes, and viruses.

[0003] Hyaluronidase inhibitors play a crucial role in the progression of various diseases. For example, overexpressed hyaluronidase degrades the extracellular matrix, facilitating tumor cell migration and proliferation; hyaluronidase homeostasis affects the integrity of the extracellular matrix (ECM) structure, wrinkle formation, and skin hydration, making it an important physiological factor related to skin aging; bacterial hyaluronidase may impair neutrophil function, promote Group B Streptococcus (GBS) invasion in non-human primates, and increase premature birth. Therefore, hyaluronidase inhibitors could provide new therapeutic strategies for human malignancies, delay skin aging, aid in the treatment of ascending GBS infection, serve as tools for studying the role of the enzyme, its substrates, and products in bacterial infection processes, and are potential candidates for combination antibacterial therapy.

[0004] Currently, hyaluronidase inhibitors are mainly derived from plant extracts (such as herb extract, asiaticoside, zephyranthes flavonoids and irisin) and microbial product extracts (such as adenosine heptapeptide). However, regardless of whether they are plant extracts or microbial extracts, batch differences can lead to large variations in the content of active ingredients, low purity, and unstable effects.

[0005] Therefore, it is essential to develop novel synthetic inhibitors of hyaluronidase that have high activity and low toxicity. Summary of the Invention

[0006] In order to solve the above-mentioned technical problems, one of the objectives of the present invention is to provide a biphenyltriazole derivative.

[0007] The technical solution adopted in this invention is as follows:

[0008] A biphenyltriazole derivative, the structural formula of which is shown in Formula I:

[0009]

[0010] Wherein, the R1 group is selected from any one of the four groups a1, a2, b1, or b2:

[0011]

[0012] The second objective of this invention is to provide a method for preparing the above-mentioned biphenyltriazole derivatives. When the R1 group is selected from a1 or a2 groups, the method for preparing the biphenyltriazole derivative is as follows: Fmoc-D-propargylglycine or Fmoc-L-propargylglycine is dissolved in tert-butanol solution, 4-acetyl-3-azidobiphenyl is added, followed by the sequential addition of copper acetate and ascorbic acid. The reaction is carried out overnight at room temperature. After the reaction is complete, the mixture is extracted with ethyl acetate, and the organic phases are combined. The organic phases are dried over anhydrous sodium sulfate and then evaporated to dryness to obtain the desired biphenyltriazole derivative.

[0013] The synthesis path can be represented as follows:

[0014]

[0015] a: 3-Bromoaniline (compound 1) was dissolved in a mixed solution of toluene, anhydrous ethanol and saturated sodium carbonate, and 4-acetylphenylboronic acid (compound 2) and bis(triphenylphosphine)palladium(II) chloride were added sequentially. The mixture was heated under reflux in a nitrogen atmosphere and reacted overnight. After the reaction was completed, the mixture was extracted and purified to obtain 4-acetyl-3-aminobiphenyl (compound 3).

[0016] b: 4-Acetyl-3-aminobiphenyl was dissolved in a mixed solution of methanol and water, perfluorobutylsulfonyl azide was added, followed by copper sulfate pentahydrate and potassium bicarbonate. The mixture was reacted overnight at room temperature. After the reaction was completed, it was purified by extraction to obtain 4-acetyl-3-azidobiphenyl (compound 4).

[0017] c: Fmoc-D-propargylglycine or Fmoc-L-propargylglycine was dissolved in tert-butanol solution, 4-acetyl-3-azidobiphenyl was added, followed by copper acetate and ascorbic acid. The reaction was carried out overnight at room temperature. After the reaction was completed, the mixture was extracted with ethyl acetate, the organic phases were combined, dried over anhydrous sodium sulfate, evaporated to dryness and purified to obtain biphenyltriazole derivatives A1 or A2.

[0018] When the R1 group is selected from b1 or b2, the preparation method of this biphenyl triazole derivative is as follows: Based on the synthesis of 4-acetyl-3-azidobiphenyl (compound 4), α,3-ynylaniline (compound 9) or 4-ynylaniline (compound 10) is dissolved in DMF solution, 4-acetyl-3-azidobiphenyl (compound 4) is added, followed by the sequential addition of ascorbic acid and tert-butyltrichloroacetylimine ester, and finally the addition of copper sulfate pentahydrate and water. The reaction is stirred at room temperature. After the reaction was completed, the mixture was extracted with ethyl acetate, the organic phases were combined, and the organic phases were dried to give a crude product. The crude product was purified to give a yellow solid 1-(3'-(4-(3-aminophenyl)-1H-1,2,3-triazol-1-yl)-[1,1'-biphenyl]-4-yl)ethyl-1-one or 1-(3'-(4-(4-aminophenyl)-1H-1,2,3-triazol-1-yl)-[1,1'-biphenyl]-4-yl)ethyl-1-one (compound 12);

[0019] b. 4,7-Dichloroquinoline was dissolved in anhydrous ethanol, compound 11 or compound 12 was added, the mixture was heated under reflux, and the mixture was dehydrated and condensed under reduced pressure and purified to give biphenyltriazole derivatives B1 or B2.

[0020] The synthesis path can be represented as follows:

[0021]

[0022] The preparation route is primarily for illustrative purposes and not for limiting the invention.

[0023] A third objective of this invention is to provide the use of the biphenyltriazole derivatives as described above and / or pharmacologically permissible salts of the biphenyltriazole derivatives as hyaluronidase inhibitors.

[0024] The fourth objective of this invention is to provide the use of the biphenyltriazole derivatives as described above and / or pharmacologically permissible salts of the biphenyltriazole derivatives in the preparation of drugs that inhibit hyaluronidase.

[0025] The fifth objective of this invention is to provide the use of the biphenyltriazole derivatives as described above and / or pharmacologically permissible salts of the biphenyltriazole derivatives in the preparation of medicaments for diseases related to hyaluronic acid.

[0026] Preferably, the hyaluronic acid-related disease is an immune-mediated inflammation.

[0027] The sixth objective of this invention is to provide the use of the biphenyltriazole derivatives as described above and / or pharmacologically permissible salts of the biphenyltriazole derivatives in products containing hyaluronic acid.

[0028] Preferably, the product is a skin care product and / or hair care product, such as skin lotion, toner, skin serum, facial cleanser, body wash, shampoo, etc.

[0029] Preferably, the pharmacologically permissible salts of the biphenyltriazole derivatives include salts formed by the biphenyltriazole derivatives and any one of an inorganic acid, an organic acid, an alkali metal, or an alkaline earth metal. The inorganic acid is any one of hydrochloric acid, sulfuric acid, or phosphoric acid. The organic acid is any one of acetic acid, maleic acid, citric acid, benzenesulfonic acid, methylbenzenesulfonic acid, fumaric acid, tartaric acid, lactic acid, or citric acid. The alkali metal is any one of lithium, sodium, or potassium. The alkaline earth metal is any one of calcium or magnesium.

[0030] The beneficial effects of this invention are as follows:

[0031] This invention provides a novel hyaluronidase inhibitor containing a biphenyltriazole structure, which has stronger activity than positive controls such as Vcpal, mild cytotoxicity, and a significant inhibitory effect on LPS-stimulated nitric oxide (NO) release from RAW264.7, indicating significant anti-inflammatory activity. Attached Figure Description

[0032] Figure 1 This study investigated the effect of biphenyltriazole derivatives on NO release from LPS-stimulated RAW264.7 macrophages. Data are the mean ± SEM values ​​from three replicates. *** indicates P < 0.001, ** indicates P < 0.01, and * indicates P < 0.05. P-values ​​were determined by a t-test between the experimental group and the model group (LPS-stimulated group). Detailed Implementation

[0033] Unless otherwise stated, the terms used herein have the meanings commonly understood by those skilled in the art.

[0034] The technical solution of the present invention will be described in more detail below with reference to embodiments:

[0035] Example 1

[0036] Synthesis of compound A1:

[0037]

[0038] 1) Synthesis of 4-acetyl-3-aminobiphenyl: 3-bromoaniline (800 mg, 4.65 mmol) was dissolved in a mixed solution of toluene (20 mL), anhydrous ethanol (10 mL), and saturated sodium carbonate (6 mL). Then, 4-acetylphenylboronic acid (762 mg, 4.65 mmol) and bis(triphenylphosphine)palladium(II) chloride (98 mg, 0.14 mmol) were added. The mixture was heated to reflux under nitrogen protection and reacted overnight. The reaction was monitored by thin-layer chromatography. After the reaction was completed, water (100 mL) was added and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate and then evaporated to dryness to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain a white solid compound 4-acetyl-3-aminobiphenyl (550 mg, 56%). 1 H NMR (400MHz, CDCl3): δ8.01(d,J=8.0Hz,2H),7.65(d,J=8.0Hz,2H),7.27-7.23(m,1H), 7.02(d,J=8.0Hz,1H),6.93(s,1H),6.73(d,J=8.0Hz,1H),3.79(s,2H),2.64(s,3H)ppm; 13 CNMR (100MHz, CDCl3): δ197.8,146.8,145.9,140.9,135.7,129.8,128.7,127.1,117.6,114.9,113.7,26.6ppm.

[0039] 2) Synthesis of 4-acetyl-3-azidobiphenyl: 4-acetyl-3-aminobiphenyl (400 mg, 1.89 mmol) was dissolved in a mixture of methanol and water. Perfluorobutylsulfonyl azide (1.2 g, 3.78 mmol) was added, followed by copper sulfate pentahydrate (47.4 mg, 0.19 mmol) and potassium bicarbonate (757.6 mg, 7.58 mmol). The reaction was carried out overnight at room temperature. The reaction was monitored by thin-layer chromatography. After the reaction was completed, water (100 mL) was added and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate and then evaporated to dryness to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain the white solid compound 4-acetyl-3-azidobiphenyl (420 mg, 93.8%). 1 H NMR (400MHz, CDCl3): δ8.03(d,J=8.0Hz,2H),7.66(d,J=8.0Hz,2H),7.45(t,J=8.0Hz, 1H),7.39(d,J=8.0Hz,1H),7.26-7.24(m,1H),7.08(d,J=12.0Hz,1H),2.64(s,3H)ppm; 13C NMR (100MHz, CDCl3): δ198.4,145.2,142.2,141.2,136.7,130.8,129.5,127.7,124.3,119.1,118.3,27.1ppm.

[0040] 3) Compound A1: Fmoc-D-propargylglycine (7) (106.7 mg, 0.45 mmol) was dissolved in a mixed solution of tert-butanol (12 mL) and water (8 mL). 4-Acetyl-3-azidobiphenyl (100 mg, 0.3 mmol) was added, followed by copper acetate (monohydrate) (29.8 mg, 0.15 mmol) and ascorbic acid (52.8 mg, 0.3 mmol). The reaction was carried out overnight at room temperature. The reaction was monitored by thin-layer chromatography. After the reaction was completed, water (100 mL) was added and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate and then evaporated to dryness to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain a white solid compound A1 (105 mg, 61.4%).

[0041] 1 H NMR (400MHz, DMSO-d6): δ8.57(s,1H),8.13(s,1H),8.03(d,J=8.0Hz,2H),7.91-7.80(m,6H),7.68- 7.61(m,3H),7.43-7.20(m,4H),6.86(s,1H),4.25-4.13(m,4H),3.16-3.10(m,2H),2.61(s,3H)ppm; 13 C NMR (100MHz, DMSO-d6): δ197.9,143.9,143.2,140.6,137.5,136.1,130.6,128.9,127.5,127 .2,127.0,126.8,124.9,121.3,120.0,119.4,118.2,109.8,65.5,46.7,26.8ppm; HR-MS[M+H] + =573.2134(calcd for C) 34 H 29 N4O5, 573.2138).

[0042] Example 2

[0043] Synthesis of compound A2:

[0044]

[0045] Based on the 4-acetyl-3-azidobiphenyl synthesized in Example 1, Fmoc-L-propargylglycine (200 mg, 0.6 mmol) was dissolved in a mixed solution of tert-butanol (16 mL) and water (12 mL), and then 4-acetyl-3-azidobiphenyl (180 mg, 0.76 mmol) was added. Cu(OAc)2·H2O (60 mg, 0.3 mmol) and ascorbic acid (105 mg, 0.6 mmol) were added sequentially. The reaction was carried out overnight at room temperature, and the reaction was monitored by thin-layer chromatography. After the reaction was completed, water (100 mL) was added and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and then evaporated to dryness to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain a white solid compound A2 (300 mg, 87.7%).

[0046] 1 H NMR (400MHz, DMSO-d6): δ8.60(s,1H),8.13(s,1H),8.03(d,J=8.0Hz,2H),7.89(d,J=8.0Hz,3H),7.81(t,J=8.0H z,3H),7.68-7.62(m,3H),7.36-7.20(m,4H),6.98(s,1H),4.25-4.12(m,4H),3.29-3.13(m,2H),2.61(s,3H)ppm; 13 CNMR (100MHz, DMSO-d6): δ197.6,155.5,145.4,143.9,143.2,140.7,140.6,137.5,136.1,130.6,128 .9,127.6,127.2,127.0,126.8,125.2,121.2,120.0,119.6,118.1,65.5,46.7,26.8ppm; HR-MS[M+H] + =573.2134(calcd for C) 34 H 29 N4O5, 573.2138).

[0047] Example 3

[0048] Synthesis of compound B1:

[0049]

[0050] 1) Based on the 4-acetyl-3-azidobiphenyl synthesized in Example 1, 3-alkynylaniline (100 mg, 0.85 mmol) was dissolved in DMF (6 mL) solution, 4-acetyl-3-azidobiphenyl (54 mg, 0.23 mmol) was added, followed by ascorbic acid (148 mg, 0.84 mmol), TBTA (22.3 mg, 0.04 mmol), and finally copper sulfate pentahydrate (21 mg, 0.08 mmol) and water. (4 mL), stir overnight at room temperature, monitor the reaction by thin-layer chromatography, add water (100 mL) after the reaction is complete and extract with ethyl acetate (100 mL × 3), combine the organic phases, dry the organic phase with anhydrous sodium sulfate and then evaporate to dryness to obtain crude product, and then purify the crude product by silica gel column chromatography to obtain yellow solid 1-(3'-(4-(3-aminophenyl)-1H-1,2,3-triazol-1-yl)-[1,1'-biphenyl]-4-yl)ethyl-1-one (65 mg, 43.9%).

[0051] 1 H NMR (400MHz, DMSO-d6): δ9.31(s,1H),8.30(s,1H),8.09(d,J=8.0Hz,2H),8.04(d,J=8.0Hz,1H),7.99(d,J=8.0Hz,2H),7.88(d,J=8.0Hz, 1H),7.74(t,J=8.0Hz,1H),7.24(s,1H),7.14(t,J=8.0Hz,1H),7.06(d,J=8.0Hz,1H),6.59(d,J=8.0Hz,1H),5.24(s,2H),2.64(s,3H)ppm; 13 C NMR (100MHz, DMSO-d6): δ197.6,149.2,148.2,143.2,140.6,137.4,136.2 ,130.7,129.5,129.0,127.3,119.7,118.3,114.0,113.2,110.6,26.8ppm.

[0052] 2) Compound B1: 4,7-Dichloroquinoline (53.7 mg, 0.25 mmol) was dissolved in anhydrous ethanol solution (20 mL), and 1-(3'-(4-(3-aminophenyl)-1H-1,2,3-triazol-1-yl)-[1,1'-biphenyl]-4-yl)ethyl-1-one (80 mg, 0.23 mmol) was added. The mixture was heated under reflux and reacted for 5 hours. After dehydration condensation under reduced pressure, the crude compound was obtained. 3 mL of petroleum ether and 3 mL of ethyl acetate solution were added to the crude compound, and the mixture was sonicated thoroughly and filtered to obtain the yellow pure compound B1 (120 mg, 95.6%).

[0053] 1 H NMR (400MHz, DMSO-d6): δ10.00(s,1H),9.54(s,1H),8.64-8.55(m,2H),8.30(s,1H),8.11-8.04(m,7H),7.92(d,J= 4.0Hz,1H),7.79-7-71(m,2H),7.64(t,J=8.0Hz,1H),7.45(d,J=8.0Hz,1H),6.99(d,J=8.0Hz,1H),2.64(s,3H)ppm; 13 CNMR (175MHz, DMSO-d6): δ197.6,146.9,143.1,140.7,137.3,136.3,131.7,129.0, 127.3,126.0,125.1,123.4,120.3,119.8,101.6,46.2,26.9,18.8ppm; HR-MS[M+H] + =516.1595(calcd forC) 31 H 23 ClN5O, 516.1591).

[0054] Example 4

[0055] Synthesis of compound B2:

[0056]

[0057] 1) Based on the 4-acetyl-3-azidobiphenyl synthesized in Example 1, 4-alkynylaniline (65 mg, 0.56 mmol) was dissolved in DMF (6 mL) solution, 4-acetyl-3-azidobiphenyl (120 mg, 0.51 mmol) was added, followed by ascorbic acid (179 mg, 1.02 mmol), TBTA (27 mg, 0.05 mmol), and finally copper sulfate pentahydrate (25 mg, 0.1 mmol) and water (4 mL). The mixture was stirred overnight at room temperature and the reaction was monitored by thin-layer chromatography. After the reaction was completed, water (100 mL) was added and the mixture was extracted with ethyl acetate (100 mL × 3). The organic phases were combined and dried over anhydrous sodium sulfate and then evaporated to dryness to obtain the crude product. The crude product was then purified by silica gel column chromatography to obtain a yellow solid 1-(3'-(4-(4-aminophenyl)-1H-1,2,3-triazol-1-yl)-[1,1'-biphenyl]-4-yl)ethyl-1-one (110 mg, 61.4%).

[0058] 1H NMR (400MHz, DMSO-d6): δ9.16(s,1H),8.27(s,1H),8.09(d,J=8.0Hz,2H),8.02-7.96(m,3H),7.86(d,J=8. 0Hz,1H),7.73(t,J=8.0Hz,1H),7.62(d,J=8.0Hz,2H),6.67(d,J=8.0Hz,2H),5.33(s,2H),2.64(s,3H)ppm; 13 C NMR (100MHz, DMSO-d6): δ197.6,149.1,148.4,143.2,140.6,137.5,136.2 ,130.7,128.9,127.2,126.8,126.4,119.5,118.1,117.4,114.0,26.8ppm.

[0059] 2) Compound B2: 4,7-Dichloroquinoline (33.6 mg, 0.17 mmol) was dissolved in anhydrous ethanol solution (20 mL), and 1-(3'-(4-(4-aminophenyl)-1H-1,2,3-triazol-1-yl)-[1,1'-biphenyl]-4-yl)ethyl-1-one (50 mg, 0.14 mmol) was added. The mixture was heated under reflux and reacted for 5 hours. After dehydration condensation under reduced pressure, the crude compound was obtained. 3 mL of petroleum ether and 3 mL of ethyl acetate solution were added dropwise, and the mixture was sonicated thoroughly and filtered to obtain the yellow pure compound B2 (71 mg, 92.2%).

[0060] 1 H NMR (400MHz, DMSO-d6): δ11.13(s,1H),9.59(s,1H),8.88(d,J=8.0Hz,1H),8.56(d,J=4.0Hz,1H),8.32(s,1H),8.06-8.04(m,6H),7.99(d,J =8.0Hz,2H),7.91(d,J=8.0Hz,1H),7.86-7.83(m,1H),7.77(t,J=8.0Hz,1H),7.64(d,J=12.0Hz,2H),6.95(d,J=4.0Hz,1H),2.64(s,3H)ppm; 13C NMR (100MHz, DMSO-d6): δ197.6,171.2,154.1,146.7,144.2,143.1,140.7,138.1,137.3,136.3,1 30.8,129.0,127.3,126.7,126.1,125.6,120.2,119.8,118.3,116.3,110.6,26.9ppm; HR-MS[M+H] + =516.1596(calcd forC) 31 H 23 ClN5O, 516.1591).

[0061] Example 5

[0062] Hyaluronidase activity inhibition test

[0063] Add 25 μL of enzyme solution (bovine testicular hyaluronidase, Sigma H3506) containing different concentrations of the test compound (see Table 1) to each well of a 96-well plate, with three replicates for each concentration. Incubate at 37°C for 10 minutes. Then add 25 μL of hyaluronic acid (0.5 mg / mL in 0.3 M PB buffer) to each well, shake for 5 minutes, incubate at 37°C for 45 minutes, and finally add 150 μL of stop solution (2.5% CTAB, 0.5 M NaOH, pH 12.5) to each well. Measure the absorbance at 600 nm using a microplate reader.

[0064] Enzyme inhibition rate = (OD 阴性对照 -OD 测试组 ) / (OD 阴性对照 -OD 空白 )×100%.

[0065] The enzyme activity inhibition rate under different concentrations of compounds was analyzed using GraphPad Prism 9 software, and the half-maximal inhibitory concentration (IC50) of the compounds was determined. 50 Values ​​are expressed as the average of three replicates ± SEM.

[0066] The results are shown in Table 1:

[0067] Table 1. Inhibitory activity of compounds against hyaluronidase

[0068]

[0069] It can be seen that the four compounds provided by this invention all have good inhibitory activity against hyaluronidase, and their inhibitory effect is better than that of the control drugs ascorbate palmitate (Vcpal) and glycyrrhizin. This indicates that the compounds provided by this invention have the potential to be developed into hyaluronidase inhibitors.

[0070] Example 6

[0071] MTT assay for compound cytotoxicity

[0072] Human umbilical vein endothelial cells (HUVEC) and human ovarian cancer (SKOV3) cell lines were cultured in DMEM complete medium (90% DMEM, 10% fetal bovine serum, 100 units / mL penicillin, 100 mg / mL streptomycin). When the cells were in good condition, they were trypsinized, collected, and the concentration adjusted. 100 μL of cell suspension was seeded into each well of a 96-well microplate, 3000 cells per well. After overnight incubation at 37°C with 5% CO2, different concentrations of the compound were added to each well, and incubation continued for 48 hours. Then, 20 μL of LTT (5 mg / mL) was added to each well, and incubation continued for 4 hours. The supernatant was discarded, and 150 μL of DMSO was added to each well, followed by shaking for 20 minutes. The optical density (OD) was read at 550 nm using a microplate reader. Three replicates were prepared for each compound concentration.

[0073] Cell proliferation inhibition rate = (OD 阴性对照 -OD 试验 ) / (OD 阴性对照 -OD 空白 )×100%.

[0074] The cell proliferation inhibition rate under different concentrations of the compound was analyzed using GraphPad Prism 9 software, and the half-maximal inhibitory concentration (IC50) of the compound was determined. 50 Values. The results are shown in Table 2:

[0075] Table 2. Cytotoxicity of the compounds

[0076]

[0077] a IC 50 The values ​​represent the concentration of the compound required to inhibit cell viability by 50%, with doxorubicin hydrochloride (Dox.) used as a positive control. The data are the mean ± SEM of three replicates.

[0078] The four compounds provided by this invention have mild cytotoxicity and have the potential to be used in drugs that inhibit hyaluronidase or products containing hyaluronic acid.

[0079] Example 7

[0080] Test for nitric oxide (NO) release

[0081] The Griess method was used to test the inhibitory effect of compounds on NO release in LPS-stimulated macrophages. RAW264.7 cells (50,000 cells / well) were seeded into 96-well plates and cultured overnight at 37°C with 5% CO2. The supernatant was carefully discarded. Compounds 6 and 14 were then diluted to different test concentrations with medium containing LPS (20 μg / mL) and added to the corresponding wells of the 96-well plates. After culturing at 37°C with 5% CO2 for 24 hours, the supernatant was collected, and the NO level in the corresponding wells was measured using the Griess method.

[0082] See results Figure 1 It can be seen that the four compounds provided by this invention have a significant inhibitory effect on the release of nitric oxide (NO) from LPS-stimulated RAW264.7, indicating that they have certain anti-inflammatory activity.

[0083] The above are merely preferred embodiments of the present invention and are not intended to limit the scope of the invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A biphenyltriazole derivative, characterized in that, The structural formula of this derivative is shown in formula (Ⅰ): （Ⅰ）； Wherein, the R1 group is selected from any one of the four groups a1, a2, b1, or b2: (a1) and (a2) and (b1) and (b2).

2. A method for preparing a biphenyltriazole derivative as described in claim 1, characterized in that, When the R1 group is selected from a1 or a2 groups, the preparation method of this biphenyltriazole derivative is as follows: Fmoc-D-propargylglycine or Fmoc-L-propargylglycine was dissolved in tert-butanol solution, 4-acetyl-3-azidobiphenyl was added, followed by copper acetate and ascorbic acid. The reaction was carried out overnight at room temperature. After the reaction was completed, the mixture was extracted with ethyl acetate, and the organic phases were combined. The organic phases were dried over anhydrous sodium sulfate and then evaporated to dryness to obtain the biphenyltriazole derivatives. When the R1 group is selected from b1 or b2 groups, the preparation method of this biphenyltriazole derivative is as follows: S1. 3-Alynylaniline or 4-Alynylaniline was dissolved in DMF solution, 4-acetyl-3-azidobiphenyl was added, followed by ascorbic acid and tert-butyltrichloroacetylimine ester, and finally copper sulfate pentahydrate and water were added. The mixture was stirred overnight at room temperature. After the reaction was completed, it was extracted with ethyl acetate. The organic phases were combined and dried to obtain the crude product. The crude product was purified to obtain a yellow solid 1-(3'-(4-(3-aminophenyl)-1H-1,2,3-triazol-1-yl)-[1,1'-biphenyl]-4-yl)ethyl-1-one or 1-(3'-(4-(4-aminophenyl)-1H-1,2,3-triazol-1-yl)-[1,1'-biphenyl]-4-yl)ethyl-1-one; S2.4,7-Dichloroquinoline was dissolved in anhydrous ethanol, and 1-(3'-(4-(3-aminophenyl)-1H-1,2,3-triazol-1-yl)-[1,1'-biphenyl]-4-yl)ethyl-1-one or 1-(3'-(4-(4-aminophenyl)-1H-1,2,3-triazol-1-yl)-[1,1'-biphenyl]-4-yl)ethyl-1-one was added. The mixture was heated under reflux and dehydrated under reduced pressure to obtain the desired biphenyltriazole derivative.

3. The production method according to claim 2, wherein The structure of the 4-acetyl-3-azidobiphenyl is as follows: The preparation method of the 4-acetyl-3-azidobiphenyl includes the following steps: S1. 3-Bromoaniline was dissolved in a mixed solution of toluene, anhydrous ethanol and saturated sodium carbonate, and 4-acetylphenylboronic acid and bis(triphenylphosphine)palladium(II) chloride were added sequentially. The mixture was heated under reflux in a nitrogen atmosphere and reacted overnight. After the reaction was completed, the mixture was extracted and purified to obtain 4-acetyl-3-aminobiphenyl. S2. 4-Acetyl-3-aminobiphenyl was dissolved in a mixed solution of methanol and water, perfluorobutylsulfonyl azide was added, followed by copper sulfate pentahydrate and potassium bicarbonate. The mixture was reacted overnight at room temperature. After the reaction was completed, it was purified by extraction to obtain 4-acetyl-3-azidobiphenyl.

4. The use of a biphenyltriazole derivative as described in claim 1 and / or a pharmacologically permissible salt of the biphenyltriazole derivative in the preparation of a medicament for inhibiting hyaluronidase.

5. The use of a biphenyltriazole derivative as described in claim 1 and / or a pharmacologically permissible salt of the biphenyltriazole derivative in the preparation of a medicament for treating diseases related to hyaluronic acid.

6. The application as described in claim 4 or 5, wherein the pharmacologically permissible salt of the biphenyltriazole derivative comprises a salt formed by the biphenyltriazole derivative and any one of an inorganic acid, an organic acid, an alkali metal, or an alkaline earth metal, wherein the inorganic acid is any one of hydrochloric acid, sulfuric acid, or phosphoric acid; the organic acid is any one of acetic acid, maleic acid, citric acid, benzenesulfonic acid, toluenesulfonic acid, fumaric acid, tartaric acid, lactic acid, or citric acid; the alkali metal is any one of lithium, sodium, or potassium; and the alkaline earth metal is any one of calcium or magnesium.

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