Rutin-derived compound, and preparation method and application thereof
By preparing compounds derived from rutin, the side effects of existing hyperuricemia drugs were solved, and the effect of efficient uric acid reduction at low doses was achieved, which had potential clinical application value.
Patent Information
- Application Number
- CN202510408607.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-04-02
- Publication Date
- 2025-07-04
AI Technical Summary
Existing hyperuricemia treatment drugs such as xanthine oxidoreductase inhibitors such as allopurinol and febulista and uric acid excretion drugs such as probenecid and benbromalon have side effects during use, and it is necessary to develop a low-toxic and highly effective natural uric acid-lowering drug.
Using rutin as a raw material, a compound derived from rutin is prepared by a multi-step reaction, including reaction with benzyl bromide, tetrabutyl ammonium bromide and palladium carbon catalysts to form a compound with a uric acid-reducing effect.
The prepared compounds have a uric acid reduction effect in a short time, and are better than benzylbromalone at low doses, and have high clinical application value and are almost free of biological toxicity.
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Abstract
Description
Technical Field
[0001] The present invention relates to a rutin derivative, in particular to a compound derived from rutin, its preparation method and application, belonging to the technical field of medicinal chemistry. Background Art
[0002] Hyperuricemia is a pathogenic factor of gout and one of the most studied diseases globally. Hyperuricemia is associated with diseases such as metabolic disorders, cardiovascular diseases, and renal dysfunction. There are two important pathogenic causes of hyperuricemia, namely excessive uric acid production and reduced elimination of uric acid and its kidneys or both. Therefore, drugs for treating hyperuricemia are classified into two categories according to their mechanisms of action: xanthine oxidoreductase inhibitors (XOIs) and uricosuric agents.
[0003] Among them, the representative drugs of XOIs are mainly allopurinol, febuxostat, topiroxostat, etc.; the main uricosuric agents are probenecid, benzbromarone, etc. However, known drugs may all produce varying degrees of side reactions after being taken by humans, such as adverse gastrointestinal reactions, renal function damage, increased risk of cardiovascular diseases, etc. Therefore, continuously developing new uric acid-lowering drugs derived from natural active substances is of great significance for reducing side effects in humans. Summary of the Invention
[0004] In order to solve the above technical problems, the present invention proposes a compound derived from rutin, its preparation method and application.
[0005] To achieve the above object, the technical solution adopted by the present invention is as follows:
[0006] A compound derived from rutin has the following general structural formula:
[0007]
[0008] The preparation method of the above-mentioned compound derived from rutin includes the following processes:
[0009] a. In the presence of a basic auxiliary agent, dissolve rutin and mix it with benzyl bromide for reaction. Concentrate the reactant under vacuum, then re-add a solvent and concentrated hydrochloric acid for reflux reaction. After the reaction is completed, cool it, and purify the precipitated product to obtain intermediate 1;
[0010] b. After dissolving intermediate 1, add an aqueous solution of tetrabutylammonium bromide and a basic auxiliary agent to it. First, stir and react for a period of time, then add N,N-dipropylsulfamoyl chloride and continue the reaction. After the reaction is completed, extract, separate and purify the organic phase to obtain intermediate 2;
[0011] c. After dissolving intermediate 2, add a palladium-carbon catalyst and stir the reaction while charging hydrogen. After the reaction is completed, filter, concentrate the filtrate and purify it to obtain the target compound, that is, compound 3.
[0012] In the above preparation method, the possible reaction process and mechanism are speculated as follows:
[0013]
[0014] Among them, Bn represents benzyl.
[0015] As some specific examples, in step a, the molar ratio of rutin to benzyl bromide is 1:(2 - 4), such as 1:2, 1:2.5, 1:3, 1:3.5, 1:4, etc.;
[0016] Preferably, the reaction conditions of rutin and benzyl bromide are stirring reaction at 50 - 80 °C for 2 - 3 h. For example, the reaction temperature is 50 °C, 60 °C, 70 °C, 80 °C, etc., and the reaction time is 2 h, 2.5 h, 3 h, etc.;
[0017] Preferably, the reaction solvent of rutin and benzyl bromide is one or more of N,N - dimethylformamide, N,N - dimethylacetamide, acetonitrile, methanol, and tetrahydrofuran.
[0018] As some specific examples, in step a, the addition amount of concentrated hydrochloric acid is 3 - 5 times the mass of rutin, such as 3 times, 3.5 times, 4 times, 4.5 times, 5 times, etc.;
[0019] Preferably, after adding concentrated hydrochloric acid in step a, the reaction conditions are stirring reaction under reflux conditions for 1 - 2 h. Preferably, the reflux solvent is one or more of ethanol, ethylene glycol, acetone, benzene, and toluene.
[0020] As some specific examples, the basic auxiliary agent is one or two of K2CO3 and Na2CO3;
[0021] Preferably, the addition amount of the basic auxiliary agent in step a is 1.5 - 2 times the molar amount of rutin, such as 1.5 times, 1.6 times, 1.7 times, 1.8 times, 1.9 times, 2 times, etc.;
[0022] Preferably, the addition amount of the basic auxiliary agent in step b is 1 - 1.5 times the molar amount of intermediate 1, such as 1 times, 1.1 times, 1.2 times, 1.3 times, 1.4 times, 1.5 times, etc.
[0023] As some specific examples, in step b, the addition amount of tetrabutylammonium bromide is 0.4 - 0.6 times the molar amount of intermediate 1, such as 0.4 times, 0.5 times, 0.6 times, etc.;
[0024] Preferably, in step b, the reaction conditions of intermediate 1 and tetrabutylammonium bromide are stirring reaction at 40 - 60 °C for 0.5 - 2 h. For example, the reaction temperature is 40 °C, 45 °C, 50 °C, 55 °C, 60 °C, etc., and the reaction time is 0.5 h, 1 h, 1.5 h, 2 h, etc.
[0025] As some specific examples, in step b, the addition amount of N,N-dipropylaminosulfonyl chloride is 2 - 3 times the molar amount of intermediate 1, such as 2 times, 2.2 times, 2.4 times, 2.5 times, 2.6 times, 2.8 times, etc.;
[0026] Preferably, after adding N,N-dipropylaminosulfonyl chloride in step b, the reaction conditions are continuous stirring reaction at 40 - 60 °C for 4 - 8 h. For example, the reaction temperature is 40 °C, 45 °C, 50 °C, 55 °C, 60 °C, etc., and the reaction time is 4 h, 5 h, 6 h, 7 h, 8 h, etc.
[0027] As some specific examples, in step b, the solvent used for dissolving intermediate 1 and the solvent used for extraction after the reaction end are each independently selected from one or more of dichloromethane, chloroform, dichloroethane, trichloroethane, dichloropropane, tetrahydrofuran, and methyl acetate;
[0028] As some specific examples, after the reaction in step b ends, first extract to obtain the organic phase, and then separate to obtain the main product, i.e., intermediate 2, by silica gel column chromatography;
[0029] Preferably, the elution solvent for the silica gel column chromatography is a dichloromethane:methanol mixed solvent with a volume ratio of 50:1.
[0030] As some specific examples, in step c, the dosage of the palladium-carbon catalyst, calculated based on the mass of metallic palladium relative to intermediate 2, is 1 - 5%, such as 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, etc.;
[0031] Preferably, the reaction conditions in step c are normal temperature, a hydrogen pressure of 0.5 - 2 MPa, and a reaction time of 4 - 8 h;
[0032] Preferably, in step c, the solvent used for dissolving intermediate 2 is an equal-volume mixed solution of ethanol and tetrahydrofuran.
[0033] The present invention also proposes an application of a compound derived from rutin as described above or a compound prepared by the method as described above in anti-hyperuricemic drugs.
[0034] Through continuous research, the present invention finds that the compound derived from rutin can not only exert the effect of reducing uric acid in a short time, but also has an even better effect on eliminating blood uric acid at a low-dose concentration than the positive drug benzbromarone, and has high clinical application value. Detailed implementation manners
[0035] The present invention will be further described below through specific embodiments. The embodiments described in the present invention are only for the purpose of explaining the present invention and do not limit the scope of the present invention.
[0036] In the following examples and comparative examples of the present invention, the main raw materials were all obtained through commercial channels on the market.
[0037] In the following Examples 1-3, the compounds derived from rutin were all synthesized by the following route:
[0038]
[0039] Example 1
[0040] a. In the presence of K2CO3 (0.41 g, 3.0 mmol), rutin (1.22 g, 2.0 mmol) was dissolved in DMF (20 mL), and benzyl bromide (0.48 mL, 4.0 mmol) was slowly added dropwise. The reaction was carried out at 60 °C for 2 h. The reaction product was concentrated in vacuo, and then ethanol (30 mL) and concentrated HCl (4 g) were added again. The temperature was raised to 85 °C and refluxed for 1 h. After the reaction was completed, it was cooled to room temperature, and the precipitate was filtered and recrystallized with ethanol to obtain a solid product, that is, intermediate 1;
[0041] 1 H NMR (500 MHz, Chloroform-d) δ 8.68 (s, 1H), 8.21 (s, 1H), 7.47 (dd, J = 9.0, 2.2 Hz, 1H), 7.42 (dq, J = 7.2, 1.1 Hz, 4H), 7.38 - 7.32 (m, 4H), 7.31 (d, J = 2.2 Hz, 1H), 7.30 - 7.24 (m, 2H), 7.06 (s, 1H), 6.89 (d, J = 9.1 Hz, 1H), 6.48 (d, J = 2.0 Hz, 1H), 6.42 (d, J = 2.2 Hz, 1H), 5.07 (t, J = 0.9 Hz, 2H), 4.95 (d, J = 1.1 Hz, 2H).
[0042] b. Dissolve intermediate 1 (0.96 g, 2.0 mmol) in CHCl3 (15 mL), add tetrabutylammonium bromide (0.32 g, 1.0 mmol) and an aqueous solution of K2CO3 (15 mL, 0.15 mol / L). First, stir the reaction at 45 °C for 2 h, then add N,N-dipropylsulfamoyl chloride (0.80 g, 4.0 mmol) and continue the reaction for 5 h. After the reaction is completed, extract with CH2Cl2 (20 mL) to obtain the organic phase. After vacuum concentration of the organic phase, purify the residue by silica gel column chromatography (volume ratio of dichloromethane:methanol = 50:1) to obtain intermediate 2;
[0043] 1 H NMR (500 MHz, Chloroform-d) δ8.47 (s, 1H), 7.54 (dd, J = 9.1, 2.3 Hz, 1H), 7.45 - 7.38 (m, 5H), 7.38 - 7.32 (m, 4H), 7.31 - 7.24 (m, 2H), 6.93 (s, 1H), 6.88 (d, J = 9.1 Hz, 1H), 6.44 (dd, J = 24.5, 2.2 Hz, 2H), 5.07 (t, J = 0.9 Hz, 2H), 4.95 (d, J = 1.1 Hz, 2H), 3.10 (t, J = 7.6 Hz, 4H), 1.62 (h, J = 7.7 Hz, 4H), 0.94 (t, J = 7.7 Hz, 6H).
[0044] c. Dissolve intermediate 2 (0.32 g, 0.50 mmol) in a mixed solvent of ethanol (5 mL) and THF (5 mL), add 10% palladium on carbon catalyst (0.06 g), and stir the reaction while charging hydrogen. The reaction conditions are room temperature, 0.5 MPa hydrogen pressure, and the reaction time is 5 h. After the reaction is completed, filter, concentrate the filtrate, and recrystallize with acetone to obtain the target compound, namely the compound derived from rutin.
[0045] 1 H NMR (500 MHz, Chloroform-d) δ8.89 (s, 1H), 7.68 (dd, J = 8.5, 2.1 Hz, 1H), 7.53 (d, J = 2.1 Hz, 1H), 7.11 (s, 1H), 6.80 (d, J = 8.5 Hz, 1H), 6.27 (d, J = 2.0 Hz, 1H), 6.21 (d, J = 1.8 Hz, 1H), 3.10 (t, J = 7.6 Hz, 5H), 1.62 (h, J = 7.6 Hz, 5H), 0.98 - 0.91 (m, 6H).
[0046] Example 2
[0047] a. In the presence of Na2CO3 (0.37 g, 3.5 mmol), rutin (1.22 g, 2.0 mmol) was dissolved in DMF (20 mL), and benzyl bromide (0.72 mL, 6.0 mmol) was slowly added dropwise. The reaction was carried out at 50 °C for 3 h. The reaction mixture was concentrated under vacuum, and then ethanol (30 mL) and concentrated HCl (5 g) were added again. The temperature was raised to 85 °C and refluxed for 2 h. After the reaction was completed, it was cooled to room temperature, and the precipitate was filtered and recrystallized with ethanol to obtain a solid product, i.e., intermediate 1;
[0048] b. Intermediate 1 (0.96 g, 2.0 mmol) was dissolved in CHCl3 (15 mL), tetrabutylammonium bromide (0.38 g, 1.2 mmol) and an aqueous solution of Na2CO3 (20 mL, 0.15 mol / L) were added. The reaction was first stirred at 50 °C for 1 h, and then N,N-dipropylsulfamoyl chloride (1.0 g, 5.0 mmol) was added and the reaction continued for 8 h. After the reaction was completed, it was extracted with CH2Cl2 (20 mL), and the organic phase was separated. After the organic phase was concentrated under vacuum, the residue was purified by silica gel column chromatography (volume ratio of dichloromethane:methanol = 50:1) to obtain intermediate 2;
[0049] c. Intermediate 2 (0.32 g, 0.50 mmol) was dissolved in a mixed solvent of ethanol (5 mL) and THF (5 mL), 10% palladium on carbon catalyst (0.1 g) was added, and hydrogen was charged and stirred for reaction. The reaction conditions were normal temperature, 1 MPa hydrogen pressure, and the reaction time was 6 h. After the reaction was completed, it was filtered, the filtrate was concentrated and recrystallized with acetone to obtain the target compound, i.e., the compound derived from rutin.
[0050] Example 3
[0051] a. In the presence of K2CO3 (0.55 g, 4.0 mmol), rutin (1.22 g, 2.0 mmol) was dissolved in DMF (20 mL), and benzyl bromide (0.96 mL, 8.0 mmol) was slowly added dropwise. The reaction was carried out at 80 °C for 2 h. The reaction mixture was concentrated under vacuum, and then ethanol (30 mL) and concentrated HCl (6 g) were added again. The temperature was raised to 85 °C and refluxed for 1 h. After the reaction was completed, it was cooled to room temperature, and the precipitate was filtered and recrystallized with ethanol to obtain a solid product, i.e., intermediate 1;
[0052] b. Dissolve intermediate 1 (0.96 g, 2.0 mmol) in CHCl₃ (15 mL), add tetrabutylammonium bromide (0.26 g, 0.8 mmol) and an aqueous solution of K₂CO₃ (18 mL, 0.15 mol / L). First, stir and react at 60 °C for 0.5 h, then add N,N-dipropylsulfamoyl chloride (1.2 g, 6.0 mmol) and continue to react for 4 h. After the reaction is completed, extract with CH₂Cl₂ (20 mL) to obtain the organic phase. After vacuum concentration of the organic phase, purify the residue by silica gel column chromatography (volume ratio of dichloromethane:methanol = 50:1) to obtain intermediate 2;
[0053] c. Dissolve intermediate 2 (0.32 g, 0.50 mmol) in a mixed solvent of ethanol (5 mL) and THF (5 mL), add 10% palladium on carbon catalyst (0.16 g), and stir and react while charging hydrogen. The reaction conditions are room temperature, 2 MPa hydrogen pressure, and the reaction time is 4 h. After the reaction is completed, filter. The filtrate is concentrated and recrystallized with acetone to obtain the target compound, that is, the compound derived from rutin.
[0054]
Cytotoxicity Test
[0055] The cytotoxicity of PC12 cells was tested by the CCK-8 method. Among them, the experimental group was the culture medium containing the compound derived from rutin prepared in Example 1; the control group was the culture medium containing rutin. The results showed that at a drug concentration of 20 - 80 μmol / L, the OD values at 450 nm of the experimental group and the control group at the same concentration had no obvious change.
[0056]
Rat Serum Uric Acid Experiment
[0057] Reagent Preparation:
[0058] (1) Preparation of 0.8 wt% CMC-Na solution: Weigh 2.4 grams of CMC-Na, first make it into a paste with a small amount of pure water, then make up the volume to 300 mL with pure water, pour it into a beaker and boil it to completely dissolve, and then pour it into a volumetric flask and make up to 300 mL with pure water.
[0059] (2) Preparation of 50 mg / ml hypoxanthine solution: Weigh 1 g of hypoxanthine and dissolve it in 20 mL of 0.8 wt% CMC-Na solution.
[0060] (3) Preparation of 0.5 mg / mL benzbromarone solution: Weigh 20 mg of benzbromarone and dissolve it in 40 mL of 0.8 wt% CMC-Na solution.
[0061] (4) Preparation of 1 mg / mL raw material control group medicinal liquid: Weigh 20 mg of rutin and 20 mg of N,N-dipropylsulfamoyl chloride and dissolve them in 40 mL of 0.8 wt% CMC-Na solution.
[0062] (5) Preparation of the experimental group liquid medicine: Dissolve the rutin-derived compound prepared in Example 1 in a 0.8 wt% CMC-Na solution to prepare low-dose experimental group liquid medicine, medium-dose experimental group liquid medicine, and high-dose experimental group liquid medicine with drug concentrations of 0.5 mg / mL, 1.0 mg / mL, and 2.0 mg / mL, respectively.
[0063] Animal experiment:
[0064] Randomly select adult male SD rats weighing 150 - 170 g / rat, and divide them into 7 groups, which are respectively recorded as the blank group, the model group, the low-dose experimental group, the medium-dose experimental group, the high-dose experimental group, the positive control group, and the raw material control group; each group consists of 10 rats.
[0065] Administer the medicine to the rats intragastrically at a dosage of 10 mL / kg per day according to their body weight for 7 consecutive days; the specific medicine administration method is as follows: both the blank group and the model group are given a 0.8 wt% CMC-Na solution; the low-dose experimental group, the medium-dose experimental group, and the high-dose experimental group are respectively given the experimental group liquid medicine with concentrations of 0.5 mg / mL, 1 mg / mL, and 2 mg / mL; the positive control group is given the positive control group liquid medicine with a concentration of 0.5 mg / mL; the raw material control group is given the raw material control group liquid medicine with a concentration of 1 mg / mL.
[0066] 30 minutes after the last administration on the 7th day, except for the blank group, other groups are intraperitoneally injected with a 50 mg / ml hypoxanthine solution to establish a hyperuricemia model, and the administration dose is 0.1 ml / 10 g. 30 minutes after the administration, immediately test the serum uric acid level of all experimental rats. The specific test methods and results are as follows:
[0067] Collect blood from the rats by enucleating the eyeballs. After the serum exudes, centrifuge at 15000 rpm for 4 minutes, aspirate the upper-layer serum into a sample tube with a lid, and prepare it to the required volume with physiological saline. Measure the serum uric acid content on a Beckman LX20 automatic biochemical analyzer. The data is expressed as the mean ± standard deviation. The test results are shown in Table 1.
[0068] Table 1. Test results of serum uric acid content
[0069] Mean ± standard deviation of blood uric acid (μmol / L) Blank group 125.42±19.82 Model group 675.41±31.52 Low-dose experimental group 554.89±27.54 Medium-dose experimental group 498.56±21.53 High-dose experimental group 429.69±22.97 Positive control group 585.27±28.46 Raw material control group 608.61±33.57
[0070] It should be noted that the above animal experiment was carried out in strict accordance with the relevant regulations of the Guide for the Care and Use of Laboratory Animals, and every effort was made to reduce the pain of the animals.
[0071] As can be seen from the above test results, the compound derived from rutin provided by the present invention can play a role in reducing uric acid within a short period of time at a dosage of 5 - 20 mg / kg, and has almost zero biological toxicity; moreover, the level of reducing uric acid at a lower dosage is significantly better than that of the positive control drug benzbromarone and the raw material control group with a mixture of rutin and N,N-dipropylsulfamoyl chloride as the liquid medicine, and has potential clinical application value for the treatment of hyperuricemia.
[0072] The above description is only a preferred embodiment of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the method of the present invention, several improvements and supplements can still be made, and these improvements and supplements should also be regarded as the protection scope of the present invention.
Claims
1. A compound derived from rutin, characterized in that, It has the following structural general formula:
2. A preparation method of a compound derived from rutin, characterized in that, It includes the following processes: ； a. In the presence of a basic auxiliary agent, dissolve rutin and then mix it with benzyl bromide for reaction. Vacuum concentrate the reactants, then re-add a solvent and concentrated hydrochloric acid for reflux reaction. After the reaction ends, cool it, and purify the precipitate to obtain Intermediate 1; b. After dissolving Intermediate 1, add an aqueous solution of tetrabutylammonium bromide and a basic auxiliary agent thereto. First, stir and react for a period of time, then add N,N-dipropylsulfamoyl chloride and continue the reaction. After the reaction ends, extract, separate and purify the organic phase to obtain Intermediate 2; c. After dissolving Intermediate 2, add a palladium-carbon catalyst and stir the reaction while charging hydrogen. After the reaction ends, filter, concentrate the filtrate and purify it to obtain the target compound, namely Compound 3.
3. The preparation method of the rutin-derived compound according to claim 2, characterized in that, In step a, the molar ratio of rutin to benzyl bromide is 1:(2 - 4); Preferably, the reaction conditions for rutin and benzyl bromide are to stir and react at 50 - 80 °C for 2 - 3 h; Preferably, the reaction solvent for rutin and benzyl bromide is one or more of N,N-dimethylformamide, N,N-dimethylacetamide, acetonitrile, methanol, and tetrahydrofuran.
4. The method for preparing a rutin-derived compound according to claim 2 or 3, characterized in that, In step a, the addition amount of concentrated hydrochloric acid is 3 - 5 times the mass of rutin; Preferably, after adding concentrated hydrochloric acid in step a, the reaction conditions are to stir and react under reflux conditions for 1 - 2 h, and the preferred reflux solvent is one or more of ethanol, ethylene glycol, acetone, benzene, and toluene.
5. The method for preparing a rutin-derived compound according to any one of claims 2-4, characterized in that, The basic auxiliary agent is one or two of K2CO3 and Na2CO3; Preferably, the addition amount of the basic auxiliary agent in step a is 1.5 - 2 times the molar amount of rutin; Preferably, the addition amount of the basic auxiliary agent in step b is 1 - 1.5 times the molar amount of Intermediate 1.
6. The preparation method of the rutin-derived compound according to any one of claims 2-5, characterized in that, In step b, the addition amount of tetrabutylammonium bromide is 0.4 - 0.6 times the molar amount of Intermediate 1; Preferably, in step b, the reaction conditions for Intermediate 1 and tetrabutylammonium bromide are to stir and react at 40 - 60 °C for 0.5 - 2 h.
7. The preparation method of the rutin-derived compound according to any one of claims 2-6, characterized in that, In step b, the addition amount of N,N-dipropylsulfamoyl chloride is 2 - 3 times the molar amount of Intermediate 1; Preferably, after adding N,N-dipropylsulfamoyl chloride in step b, the reaction conditions are to continue stirring and reacting at 40 - 60 °C for 4 - 8 h.
8. The method for preparing a rutin-derived compound according to any one of claims 2-7, characterized in that, After the reaction in step b ends, first extract to obtain an organic phase, and then separate the main product, namely Intermediate 2, by silica gel column chromatography; Preferably, the elution solvent for the silica gel column chromatography is a dichloromethane:methanol mixed solvent with a volume ratio of 50:
1.
9. The preparation method of the rutin-derived compound according to any one of claims 2-8, characterized in that, In step c, the dosage of the palladium-carbon catalyst is 1 - 5% based on the mass of metallic palladium relative to Intermediate 2; Preferably, the reaction conditions in step c are normal temperature and a hydrogen pressure of 0.5 - 2 MPa.
10. Use of the compound derived from rutin according to claim 1 or the compound prepared by the method according to any one of claims 2 - 9 in anti-hyperuricemic drugs.