Application of benzoyl paeoniflorin compound in preparation of medicine for treating hyperuricemia

Abstract

The invention belongs to the field of medicines or health care products, and particularly relates to new application of benzoyl paeoniflorin compounds in preparation of medicines for treating hyperuricemia. The research finds that the benzoyl paeoniflorin compound separated from the paeoniflorin water extract has uric acid reducing activity, and can be used for treating hyperuricemia and gout or gout complications caused by hyperuricemia.

Description

Technical Field

[0001] The invention belongs to the field of medicines or health products, and specifically relates to a new use of benzoylpeonidin compounds in the preparation of a drug for treating hyperuricemia. Background Art

[0002] Uric acid is the final metabolite of purine compounds in humans, and purine metabolism disorders lead to hyperuricemia. Under normal purine diet conditions, fasting blood uric acid levels on two different days are higher than 416 μmol / L for men and higher than 360 μmol / L for women, which is called hyperuricemia. Gout is a crystal-related arthropathy caused by the deposition of monosodium urate (MSU), which is directly related to hyperuricemia caused by purine metabolism disorders and (or) decreased uric acid excretion. The main clinical manifestations are hyperuricemia, repeated attacks of gouty acute arthritis, gouty chronic arthritis and tophi, gouty nephropathy, and renal uric acid stones. In severe cases, joint disability and renal insufficiency may occur. Currently, the treatment of hyperuricemia, gout and gout complications is mainly through controlling the uric acid in the blood. Its mechanism of action mainly includes the following two aspects: (1) effectively inhibiting the formation of uric acid by inhibiting the activity of xanthine oxidase (XO). Representative drugs include allopurinol and febuxostat. (2) promoting the excretion of uric acid. Representative drugs include probenecid and benzbromarone.

[0003] Modern research shows that the main chemical components of the water-soluble part of peony are various monoglycoside compounds such as paeoniflorin, which is the main medicinal part of red peony root. The existing biological activities reported are as follows: 1. Immunomodulation: such as inhibiting inflammatory factors; inflammatory injury diseases (arthritis, atherosclerosis, etc.); anti-inflammatory, treatment of rheumatoid arthritis; immunosuppressant; 2. Blood system: treatment or prevention of chronic peripheral vascular occlusive diseases, proliferative vascular lesions, blood tonic, and blood coagulation; 3. Regulating smooth muscle: treatment of esophageal achalasia; erectile dysfunction; promoting postoperative intestinal function recovery to prevent intestinal pinching; relieving local pain; 4. Liver protection; 5. Acting on the nervous system: anti-depression; improving menopausal syndrome; improving sleep, etc. There is no report on the use of paeoniflorin compounds for hyperuricemia. Summary of the invention

[0004] The present invention provides a new use of a benzoylpeonidin compound in the preparation of a method for treating hyperuricemia. The present invention is achieved through the following technical solutions: The present invention provides the use of benzoylpeonidin compounds represented by formula (I) and pharmaceutically acceptable salts, esters, prodrugs or solvates thereof in the preparation of medicines having uric acid lowering effects. Wherein, R1, R2, and R3 are independently selected from H, OH, and OCH3.

[0005] Preferably, the use of the benzoylpeonidin compounds represented by the above formula (I) and their pharmaceutically acceptable salts, esters, prodrugs or solvates in the preparation of medicines having a uric acid-lowering effect is characterized in that the benzoylpeonidin compounds represented by formula (I) are selected from: Compound 1: R1 is OH, R2 is H, and R3 is H; Compound 2: R1, R2, and R3 are all H; Compound 3: R1, R2 are H, and R3 is OH; Compound 4: R1 is OCH3, R2 is OH, and R3 is H.

[0006] Further preferred is the use of the benzoylpeonidin compounds represented by the above formula (I) and pharmaceutically acceptable salts, esters, prodrugs or solvates thereof in the preparation of medicines having a uric acid-lowering effect, characterized in that the benzoylpeonidin compounds represented by the formula (I) and pharmaceutically acceptable salts, esters, prodrugs or solvates thereof are prepared according to conventional processes and with conventional excipients to form clinically acceptable tablets, capsules, powders, mixtures, pills, granules, syrups, patches, suppositories, aerosols, ointments or injections.

[0007] The present invention also provides the use of benzoylpeonidin compounds represented by formula (I) and pharmaceutically acceptable salts, esters, prodrugs or solvates thereof in the preparation of medicines for treating gout. Wherein, R1, R2, and R3 are independently selected from H, OH, and OCH3.

[0008] Preferably, the use of the benzoylpeonidin compounds represented by the above-mentioned formula (I) and their pharmaceutically acceptable salts, esters, prodrugs or solvates in the preparation of medicines for treating gout is characterized in that the benzoylpeonidin compounds represented by formula (I) are selected from: Compound 1: R1 is OH, R2 is H, and R3 is H; Compound 2: R1, R2, and R3 are all H; Compound 3: R1, R2 are H, and R3 is OH; Compound 4: R1 is OCH3, R2 is OH, and R3 is H.

[0009] Further preferred is the use of the benzoylpeonidin compounds represented by the above formula (I) and pharmaceutically acceptable salts, esters, prodrugs or solvates thereof in the preparation of medicines for treating gout, characterized in that the benzoylpeonidin compounds represented by formula (I) and pharmaceutically acceptable salts, esters, prodrugs or solvates thereof are prepared into clinically acceptable tablets, capsules, powders, mixtures, pills, granules, syrups, plasters, suppositories, aerosols, ointments or injections according to conventional processes and with the addition of conventional excipients.

[0010] The technical solution of the present invention has the following advantages: This study found that benzoylpeonidin compounds extracted from peony medicinal materials have xanthine oxidase inhibitory effects and uric acid-lowering activity, and can be used to treat hyperuricemia and gout or gout complications caused by hyperuricemia. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] Figure 1 Benzoylpeonidin compounds formula (Ⅰ) Structural formula Figure 2 Compound 1 structure Figure 3 Compound 2 structure Figure 4 Compound 3 structure Figure 5 Compound 4 structure Specific embodiments

[0012] Example 1 Preparation of Benzoylpeonidin Compounds Take 20 kg of commercially available red peony root, crush it, add pure water and heat it under reflux for extraction 3 times, the solid-liquid ratio is 1:8, each time for 45 minutes, filter, combine the three extracts, and concentrate under reduced pressure to obtain an extract. The extract is separated by D101 macroporous adsorption resin, and water, 30% ethanol, 60% ethanol, and 95% ethanol are used for elution in sequence. After the eluents are concentrated under reduced pressure, the 30% ethanol elution component is separated by MCI column (column packing model CHP20 / P30), and water, 30%, 60%, and 95% ethanol aqueous solution are used for elution in sequence. The 60% elution component is concentrated under reduced pressure to dryness and marked as WA3-E60. The 60% ethanol elution component is separated by MCI column (column packing model CHP20 / P30), and water, 50%, 70%, and 95% ethanol aqueous solution are used for elution in sequence. The 50% elution component is concentrated under reduced pressure to dryness and marked as WA6-E50. WA3-E60 was separated and purified by reverse C18 column chromatography, and eluted with methanol-water solutions with volume concentrations of 15%, 40%, and 70% in sequence. After collecting the 40% and methanol-water elution fractions, the fractions were concentrated under reduced pressure and purified by Agilent SD-1 preparative liquid chromatography. A reverse C8 column was used for isocratic elution with 30% acetonitrile-water solution. According to the liquid detection spectrum, compounds 1 and 2 were obtained in sequence. WA6-E50 was separated and purified by reverse C18 column chromatography, and eluted with methanol-water solutions with volume concentrations of 20%, 50%, and 80% in sequence. After collecting the 50% methanol-water elution fractions, the fractions were concentrated under reduced pressure and purified by Agilent SD-1 preparative liquid chromatography. A reverse C8 column was used for isocratic elution with 40% acetonitrile-water solution. According to the liquid detection spectrum, compounds 3 and 4 were obtained in sequence, with HPLC purity ≥98%. The specific compound structures are shown in Figure 1 As shown, the structural confirmation data of the compounds 1H NMR and 13C NMR refer to the following literature: Compound 1 and Compound 3 refer to Ding L, Jiang Z, Liu Y, et al. Monoterpenoid inhibitors of NO production from Paeonia suffruticosa[J]. Fitoterapia, 2012, 83(8):1598-1603. Compound 2 refer to Cheng Y, Peng C, Zhang H, et al. Structural Characterization of an Artefact and Simultaneous Quantification of Two Monoterpenes and Their Artefacts of Isolation in White-Peony Root[J]. Helvetica Chimica Acta, 2010, 93:565-572. Compound 4 refer to Wu SH, Luo XD, Ma YB, et al. Monoterpenoid derivatives from Paeonia delavayi.[J]. Journal of Asian natural products research, 2002, 4(2): 135-140. Example 2 In vitro inhibition experiment of benzoylpeonidin compounds on xanthine oxidase 1. Experimental methods 1.1 Solution preparation Preparation of phosphate buffered saline (PB) Prepare K2HPO4 (weigh 5.23g K2HPO4 and dissolve it in 400mL water) and KH2PO4 solution (weigh 1.02g KH2PO4 and dissolve it in 100mL water) in advance, then accurately measure 324mL K2HPO4 and 76mL KH2PO4, mix them evenly, and you will get 50mM, pH7.5 phosphate buffer PB as the test solution for the blank control group.

[0013] Preparation of allopurinol solution Weigh 5 mg of allopurinol and dissolve it in 1 mL of DMSO to make a stock solution. After it is completely dissolved, dilute it with PB to 1.5 μg / mL and 10 μg / mL, and store it in aliquots of 1 mL / tube in a -20°C refrigerator as the test solution for the positive control group.

[0014] Take 1 tube for each experiment without dilution and use directly after thawing.

[0015] Among them, the allopurinol solution with a concentration of 1.5 μg / mL was used as the allopurinol low-dose group, and the allopurinol solution with a concentration of 10 μg / mL was used as the allopurinol high-dose group.

[0016] Preparation of xanthine oxidase solution According to the enzyme amount and volume indicated on the xanthine oxidase product package, dilute it with PB to 100U / L. After preparation, divide it into 8mL / tube and store it in a -20℃ environment.

[0017] Take one tube each time and dilute it to 25 U / L with PB for later use.

[0018] Preparation of substrate xanthine solution Accurately weigh 4.56 mg of xanthine, place in a 50 mL volumetric flask, add 150 µL of 1 M sodium hydroxide, 30 mL of PB, and sonicate until the xanthine is completely dissolved and becomes clear and transparent, then dilute to the mark with PB to obtain a 600 µM xanthine solution.

[0019] Preparation of test solution 20 mg of compounds 1, 2, 3, and 4 prepared in Example 1 and a commercially available paeoniflorin reference substance (HPLC purity ≥ 98%) were weighed and added to a 1.5 mL centrifuge tube, and then 1 mL of DMSO was added and ultrasonically dissolved to form a mother solution. 50 μL of the mother solution and 950 μL of PB were drawn and mixed to obtain a solution of the test drug.

[0020] 1.2 Reaction and termination of enzyme and substrate In the experimental group, 400 μL of xanthine oxidase (XOD) solution and 200 μL of test solution were added to the 1.5 mL centrifuge tube in sequence; in the blank control group, 400 μL of xanthine oxidase (XOD) solution and 200 μL of test solution of the blank control group were added to the 1.5 mL centrifuge tube in sequence; in the positive control group, 400 μL of xanthine oxidase (XOD) solution and 200 μL of test solution of the positive control group, low-dose group and high-dose group were added to the 1.5 mL centrifuge tube in sequence; then, each group performed the following experimental operations: incubated in a 37°C water bath for 10 min, 400 μL of xanthine (XAN) solution was added to start the reaction, after reacting in a 37°C water bath for 10 min, 80 μL of 1 M hydrochloric acid was added to terminate the reaction, and then 70-75 μL of 1 M NaOH solution was added to adjust the pH to neutral.

[0021] 1.3 Centrifugation and machine testing After the treated test solution of each group was centrifuged at 12000 rpm for 10 min at room temperature, 600 μL of the supernatant was aspirated into a liquid phase bottle, and the uric acid value of each group of test solution was detected by HPLC.

[0022] 2. Experimental data detection and processing The xanthine oxidase inhibition rate was calculated according to the following formula: inhibition rate = [(uric acid value of the blank control group - uric acid value of the test solution) / uric acid value of the blank control group] * 100.

[0023] 3. Experimental results The specific experimental results of the inhibition rate of each benzoylpeonidin compound on xanthine oxidase are shown in Table 1.

[0024] Table 1 Specific experimental results of the inhibition rate of compounds 1-4 on xanthine oxidase Group Inhibition rate (%) Positive control low dose group 39.6% Positive control high dose group 93.3% Paeoniflorin 36.1% Compound 1 89.2% Compound 2 92.5% Compound 3 86.3% Compound 4 90.7% From the inhibition rate results in Table 1, it can be seen that benzoylpeonidin compounds 1-4 all have a certain xanthine oxidase inhibitory effect, and the XOD inhibition rate is significantly higher than that of the paeoniflorin group. It can be seen that the benzoylpeonidin compounds of the present invention have a benzoyl structure in their structure compared with paeoniflorin, which enhances their inhibitory effect on xanthine oxidase, and the inhibition rates of compounds 1-4 on xanthine oxidase are higher than those of the positive control low-dose group, which have significant xanthine oxidase inhibitory effects and can be used as potential uric acid-lowering drugs.

Claims

1. The use of the benzoylpeonidin compounds shown in formula (I) of Figure 1 and their pharmaceutically acceptable salts, esters, prodrugs or solvates in the preparation of uric acid-lowering drugs, wherein: R1, R2, R3 are independently selected from H, OH, OCH3.

2. The use of the benzoylpeonidin compounds represented by formula (I) according to claim 1 and their pharmaceutically acceptable salts, esters, prodrugs or solvates in the preparation of uric acid-lowering drugs, characterized in that: The benzoylpeonidin compounds represented by formula (I) are selected from: Compound 1: R1 is OH, R2 is H, and R3 is H; Compound 2: R1, R2, and R3 are all H; Compound 3: R1, R2 are H, and R3 is OH; Compound 4: R1 is OCH3, R2 is OH, and R3 is H.

3. Use of the benzoylpeonidin compounds of formula (I) and pharmaceutically acceptable salts, esters, prodrugs or solvates thereof according to claim 1 or 2 in the preparation of a drug having a uric acid-lowering effect, characterized in that: The benzoylpeonidin compounds represented by formula (I) and pharmaceutically acceptable salts, esters, prodrugs or solvates thereof are prepared into clinically acceptable tablets, capsules, powders, mixtures, pills, granules, syrups, patches, suppositories, aerosols, ointments or injections by conventional processes and adding conventional excipients.

4. Use of benzoylpeonidin compounds represented by formula (I) and pharmaceutically acceptable salts, esters, prodrugs or solvates thereof in the preparation of drugs for treating gout, wherein: R1, R2, R3 are independently selected from H, OH, OCH3.

5. The use of the benzoylpeonidin compound represented by formula (I) and its pharmaceutically acceptable salt, ester, prodrug or solvate in the preparation of a drug for treating gout according to claim 4, characterized in that: The benzoylpeonidin compounds represented by formula (I) are selected from: Compound 1: R1 is OH, R2 is H, and R3 is H; Compound 2: R1, R2, and R3 are all H; Compound 3: R1, R2 are H, and R3 is OH; Compound 4: R1 is OCH3, R2 is OH, and R3 is H.

6. Use of the benzoylpeonidin compounds of formula (I) and pharmaceutically acceptable salts, esters, prodrugs or solvates thereof in the preparation of a drug for treating gout according to claim 4 or 5, characterized in that: The benzoylpeonidin compounds represented by formula (I) and pharmaceutically acceptable salts, esters, prodrugs or solvates thereof are prepared into clinically acceptable tablets, capsules, powders, mixtures, pills, granules, syrups, patches, suppositories, aerosols, ointments or injections by conventional processes and adding conventional excipients.

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