Use of caraphenol a in the preparation of xanthine oxidase inhibiting medicaments

Abstract

The application belongs to the technical field of effective components of Chinese medicinal materials, and particularly relates to application of Caraphenol A in preparation of xanthine oxidase inhibiting drugs. 42 H 28 O9, the Caraphenol A can be prepared into drug dosage forms suitable for clinical use, including injection, granules, capsules, tablets, powders, drop pills, sustained-release preparations, and has the action of significantly inhibiting xanthine oxidase activity.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of application of effective components of traditional Chinese medicinal materials, and particularly relates to application of Caraphenol A in preparation of xanthine oxidase (XO) inhibiting drugs. BACKGROUND

[0002] Gout is an inflammatory arthritis caused by deposition of urate crystals in joints due to persistent hyperuricemia. Hyperuricemia is generally considered a hallmark of gout, and statistics show that about 5%-12% of hyperuricemia eventually develops into gout. When the blood uric acid level exceeds its saturation in blood or tissues, crystals are formed in joints and other parts, local inflammation and tissue damage occur; the typical feature of hyperuricemia is the increase of uric acid level in the body, and controlling the increase of uric acid level is an important way to treat hyperuricemia and gout. The goal of clinical treatment of gout is to reduce the serum uric acid (SUA) level of patients to below 360 mmol / L, and for gout patients with complications, the SUA level needs to be controlled below 300 mmol / L, so as to effectively control the onset of gout.

[0003] The final metabolite of purine substances in the human body is uric acid, and XO is a key enzyme for catalyzing the generation of uric acid from xanthine and hypoxanthine in the human body. Xanthine oxidase inhibitors are the main treatment means for reducing the blood uric acid level of gout patients; at the same time, since XO inhibitors usually have smaller side effects than uric acid excretion promoting drugs and anti-inflammatory drugs, xanthine oxidase inhibitors are of great significance in the treatment of gout. The uric acid-lowering drug allopurinol has been used in clinical treatment since 1963 and has been used as a first-line anti-gout drug, but due to its adverse reactions such as allergy, liver and kidney damage, and bone marrow suppression, the total incidence of side effects is 5-20%, which greatly limits its clinical use; in 2009, the XO selective inhibitor febuxostat was approved for marketing by the US FDA, and its inhibition of XO activity is further improved, and the effect is better than that of allopurinol, but it still has adverse reactions such as stomachache, diarrhea, liver damage and muscle pain.

[0004] Therefore, further research and development of xanthine oxidase inhibitors to find more effective and reasonable anti-gout drugs is also an important task for current drug researchers. SUMMARY

[0005] The application provides application of Caraphenol A in preparation of xanthine oxidase inhibiting drugs.

[0006] Specifically, the application is achieved by the following technical solutions.

[0007] The application of Caraphenol A in preparing xanthine oxidase inhibitor drugs, wherein the xanthine oxidase inhibitor drugs are prepared from Caraphenol A as an active ingredient and other pharmaceutical excipients or carriers.

[0008] The Caraphenol A is a stilbene trimer with a molecular formula of C 42 H 28 O9, and a chemical name of (2S, 2aS, 7R, 7aR)-2, 7, 12-Tris (4-hydroxyphenyl)-2, 2a, 7, 7a-tetrahydrobis [1] benzofuro [3', 4': 4, 5, 6; 3", 4": 7, 8, 9] cyclonona [1, 2, 3-cd] [1] benzofuran-4, 9, 14-triol.

[0009] The structural formula of the Caraphenol A is as follows:

[0010]

[0011] The Caraphenol A is directly extracted from natural medicines (such as Genista Root) or prepared by chemical synthesis.

[0012] The xanthine oxidase inhibition is manifested by reducing the generation of uric acid in vivo by inhibiting the activity of xanthine oxidase, and preventing and treating diseases related to the activity of xanthine oxidase, such as hyperuricemia, gout, diabetic nephropathy, cardiovascular diseases, etc.

[0013] The xanthine oxidase inhibitor drugs are oral agents or injection agents.

[0014] Further preferably, the oral agents are any one or more of granules, capsules, tablets, powders, dripping pills and sustained-release agents.

[0015] Beneficial effects:

[0016] The Caraphenol A of the application is a monomer compound separated from Genista Root medicinal materials with a history of clinical application for many years, which is safe and effective, is a pure natural ingredient, is an effective component of Genista Root for resisting gout, and has no related side effects reported. The Caraphenol A still has a significant inhibitory effect on xanthine oxidase activity at a low concentration, and its IC 50 (23.6 mol / L) is obviously better than that of the clinical drug allopurinol (IC 50 : 71.7 mol / L)

[0017] The application is to use Caraphenol A as an active ingredient to be combined with other pharmaceutical excipients or carriers to prepare a pharmaceutical preparation, which can not only treat gout, but also avoid the occurrence of adverse reactions such as stomachache, diarrhea, liver damage and muscle pain.

[0018] The Caraphenol A has good compatibility, can be adapted to various excipients, and is suitable for the preparation process of various dosage forms such as injection, granules, capsules, tablets, powders, drop pills, sustained-release preparations and the like, which greatly widens the medication route. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 Inhibition of Caraphenol A with different concentrations on xanthine oxidase. DETAILED DESCRIPTION

[0020] The specific embodiments of the application are further described in detail below, but the application is not limited to these embodiments, and any improvement or replacement in the basic spirit of the embodiments still belongs to the scope of protection claimed by the application.

[0021] Example 1

[0022] The application of Caraphenol A in preparing xanthine oxidase inhibiting drugs, wherein the xanthine oxidase inhibition is to reduce the generation of uric acid in vivo by inhibiting the activity of xanthine oxidase, and to prevent and treat diseases related to the activity of xanthine oxidase such as hyperuricemia, gout, diabetic nephropathy, cardiovascular disease and the like;

[0023] The xanthine oxidase inhibiting drug is prepared by using Caraphenol A as an active ingredient and other pharmaceutical excipients or carriers;

[0024] The Caraphenol A is a stilbene trimer, and its molecular formula is C 42 H 28 O9, and its chemical name is (2S, 2aS, 7R, 7aR)-2, 7, 12-Tris (4-hydroxyphenyl)-2, 2a, 7, 7a-tetrahydrobis [1] benzofuro [3', 4': 4, 5, 6; 3", 4": 7, 8, 9] cyclonona [1, 2, 3-cd] [1] benzofuran-4, 9, 14-triol.

[0025] The structural formula of the Caraphenol A is as follows:

[0026]

[0027] The Caraphenol A is directly extracted from the plant Radix Triclisiae Fortunei;

[0028] The pharmaceutical dosage form of the xanthine oxidase inhibitor drug is oral or injection, specifically including but not limited to the following dosage forms: injection, granule, capsule, tablet, powder, dripping pill, sustained-release preparation.

[0029] Example 1: Test of XO inhibitory activity

[0030] At 25℃, using a 96-well plate, the total reaction volume is 200 μL, first add 50 μL of enzyme solution (the final concentration of the reaction is 0.05 U / mL), 50 μL of sample solution, incubate at 25℃ for 15 min, then add 50 μL of xanthine substrate solution (the final concentration is 150 μmol / L) to start the reaction. Incubate at 25℃ for 20 min, then add 50 μL of 1 mol / L hydrochloric acid solution to terminate the reaction. Detect the absorbance value at 290 nm. Take the difference between this value and the OD value at 0 min of incubation as the final detection result; detect the OD value of the blank sample (i.e. without the test drug, replace the sample solution with 5% DMSO in PBS to determine the maximum reaction activity of the enzyme) by the same method, and calculate the inhibition rate of each extract according to the following formula: inhibition rate (%) = (1-test sample average OD value / blank sample average OD value) x 100%. Data analysis is performed using Graphpad prism 6.0 software. Calculate the drug concentration at which half of the xanthine oxidase is inhibited, i.e. IC 50 Use allopurinol as a positive drug; the activity results are shown in Table 1;

[0031] The preparation of the enzyme solution: weigh an appropriate amount of XO, dissolve in 1 mL of water to obtain a XO stock solution with a concentration of 100 U / mL; divide the above stock solution into 10 mL EPP tubes at 50 μL per tube and store in a -80℃ refrigerator. Take an appropriate amount of the above stock solution and dilute with PBS to prepare a XO solution with a concentration of 0.2 U / mL. After the solution is prepared, it is immediately stored in a 0℃ refrigerator.

[0032] Preparation of sample solution: accurately weigh an appropriate amount of Caraphenol A, dissolve in DMSO, and then dilute with PBS to prepare a sample stock solution with a mass concentration of 1000 μg / mL. Use the double dilution method to dilute the above stock solution step by step with DMSO-containing PBS as the solvent to prepare sample solutions with mass concentrations of 1000, 500, 250, 125, 62.5, 31.3, 15.6 μg / mL, respectively, and the DMSO content is 1%.

[0033] Preparation of allopurinol control solution: accurately weigh allopurinol, dissolve it with DMSO, dilute with PBS to prepare allopurinol stock solution with a mass concentration of 500 μg / mL. The stock solution is diluted with PBS containing DMSO by two-fold dilution to prepare control solutions with mass concentrations of 500, 250, 125, 62.5, 31.3, 15.63, 7.82, 3.91 μg / mL, respectively, and the DMSO content is 1%.

[0034] Preparation of xanthine substrate solution: weigh xanthine, add PBS, adjust pH to 7.5 with NaOH and HCl solution at room temperature, and ultrasonicate to dissolve, to prepare xanthine substrate solutions with concentrations of 1200, 1000, 800, 600, 400 μmol / L, respectively.

[0035] Table 1 Inhibition rate of Caraphenol A at different concentrations on XO and half-inhibitory concentration (IC 50 )

[0036]

[0037] As shown in Table 1, compared with the IC 50 (71.7 μmol / L) and lower inhibition rate (8.3% and 4.2% at 12.5 and 6.25 μmol / L, respectively) of the positive drug allopurinol, the IC 50 of the Caraphenol A protected by the present application is 23.6 μmol / L, and it still has a high inhibition rate (42.1% and 20.1% at 12.5 and 6.25 μmol / L, respectively) at low concentrations, and its inhibition effect on xanthine oxidase is outstanding, which is obviously superior to allopurinol.

Claims

1. Use of Caraphenol A for the preparation of a medicament for preventing and treating hyperuricemia and gout by inhibiting the activity of xanthine oxidase, characterized in that, The Caraphenol A is stilbene trimer, whose molecular formula is C 42 H 28 O9, chemical name is (2S, 2aS, 7R, 7aR)-2, 7, 12-Tris (4-hydroxyphenyl)-2, 2a, 7, 7a-tetrahydrobis[1] benzofuro[3', 4':4, 5, 6; 3", 4":7, 8, 9] cyclonona[1, 2, 3-cd][1] benzofuran-4, 9, 14-triol; the structural formula of the Caraphenol A is as follows: The medicament is made of active ingredient Caraphenol A and other pharmaceutical excipients.

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