CN118812476B

CN118812476B - A compound of quercetin derived from blueberry tree and probenecid, as well as its preparation method and application

Info

Publication number: CN118812476B
Application number: CN202410791720.8A
Authority: CN
Inventors: 王磊; 周鹏; 张敏; 章晨润; 朱海亮
Original assignee: Jiangsu Forestry Academy
Current assignee: Jiangsu Forestry Academy
Priority date: 2024-06-19
Filing date: 2024-06-19
Publication date: 2025-09-12
Anticipated expiration: 2044-06-19
Also published as: CN118812476A

Abstract

本发明公开了一种乌饭树来源槲皮素接丙磺舒的化合物及其制备方法和应用。本发明的一种乌饭树来源槲皮素接丙磺舒的化合物，乌饭树来源槲皮素接丙磺舒的化合物的结构式如式(I)所示：本发明通过一系列化学反应步骤合成，包括溶剂溶解、加热搅拌、真空浓缩、萃取和硅胶柱层析等制得乌饭树来源槲皮素接丙磺舒的化合物。本发明的乌饭树来源槲皮素接丙磺舒的化合物，这种衍生物通过结构改造，具有更好的生物活性和治疗效果。本发明的化合物对降低实验鼠的血尿酸、血糖、血清总胆固醇和提高血清总蛋白水平具有显著效果。The present invention discloses a compound comprising quercetin derived from the Japanese blueberry tree and probenecid, as well as a preparation method and application thereof. The compound comprising quercetin derived from the Japanese blueberry tree and probenecid of the present invention has a structural formula as shown in formula (I): The present invention synthesizes a compound comprising quercetin derived from the Japanese blueberry tree and probenecid through a series of chemical reaction steps, including solvent dissolution, heating and stirring, vacuum concentration, extraction, and silica gel column chromatography. The compound comprises quercetin derived from the Japanese blueberry tree and probenecid, and this derivative has been structurally modified to exhibit enhanced biological activity and therapeutic efficacy. The compound has a significant effect on lowering blood uric acid, blood sugar, and serum total cholesterol, and increasing serum total protein levels in experimental mice.

Description

Vaccinium bracteatum-derived quercetin probenecid compound, and preparation method and application thereof

Technical Field

The invention relates to the technical field of chemistry, in particular to a compound of quercetin probenecid from blueberry, and a preparation method and application thereof.

Background

Uric acid is the final product of purine catabolism and is physiologically excreted by urine. Endogenous uric acid is about 80% of serum total uric acid (SUA), the remainder being from dietary purines. Most bacteria and mammals catabolize uric acid by uricase mediated metabolism to obtain water soluble allantoin. However, humans lack uricase, uric acid being excreted primarily through renal excretion. Thus, uric acid hypersecretion or renal hypovolemia can lead to hyperuricemia, which is generally defined as serum uric acid levels >6.8mg/dL. Hyperuricemia may be asymptomatic, but when serum total uric acid concentration exceeds 6.8mg/dL, monosodium urate may crystallize and deposit in joints or surrounding tissues—this is why gout is a serious disease affecting millions of people, especially adult males. Hyperuricemia is also associated with a range of chronic diseases including hypertension, diabetes, metabolic syndrome, and renal and cardiovascular diseases. Obesity and excessive intake of meat and alcoholic beverages are considered causative factors of hyperuricemia and/or gout. 98% of uric acid in the body is in the form of sodium salt. Gouty arthritis is caused by accumulation of sodium urate (MSU) crystals in joints and other tissues, causing lesions and inflammatory reactions. In gout, excessive uric acid is deposited in the joints and phagocytized by macrophages, resulting in the release of inflammatory factors, which in turn lead to acute synovitis and destruction of articular cartilage. Therefore, it is important to control the occurrence of local inflammatory reactions.

Vaccinium bracteatum (school name: vaccinium bracteatum thunder.) belonging to genus Vaccinium of family Ericaceae is a perennial shrub widely distributed in east Asian. Oriental blueberry is known as a fruit, namely, a Chinese blueberry seed, and the fruit is unique in color and luster, high in nutritive value and has a long application history in traditional medicine. The leaves, bark and fruits of vaccinium bracteatum are rich in a variety of bioactive components, including a large amount of quercetin. Quercetin is a polyphenol flavonol compound, and has remarkable biological activity, including anti-inflammatory, antioxidant, antitumor and blood pressure lowering effects.

Quercetin (Quercetin) has molecular formula of C ₁₅H₁₀O₇, and is a flavonoid compound widely existing in plant kingdom. It is distributed in various vegetables, fruits and herbs, especially in onion, apple, tea and blueberry, with high content. Quercetin is paid attention to because of its strong antioxidant properties, and can scavenge free radicals in vivo, reduce oxidative stress, and thereby protect cells from injury. In addition, quercetin has anti-inflammatory effect, and can inhibit release of inflammatory mediators and reduce inflammatory reaction. Studies have shown that quercetin can lower blood pressure through a variety of mechanisms, including blocking Angiotensin Converting Enzyme (ACE) and dilating vascular smooth muscle.

The quercetin in the blueberry is rich in content, so that the blueberry becomes a medicinal plant resource with development potential. Through modern extraction and purification technology, quercetin with high purity can be obtained from blueberry, and medicines for treating various diseases are further developed. The invention is based on the characteristics of quercetin in blueberry, and combines the quercetin with probenecid by a chemical synthesis method to prepare a novel compound. The compound not only maintains the bioactivity of quercetin, but also enhances the therapeutic effect thereof through structural modification, and particularly shows remarkable effect in reducing blood uric acid, blood sugar and serum total cholesterol.

In the field of hyperuricemia treatment, the traditional therapeutic drugs often have the problems of larger side effects, poor patient compliance and the like.

At present, a compound of quercetin probenecid derived from blueberry with small side effect, a preparation method and application thereof are lacking.

Disclosure of Invention

The invention aims to provide a compound of quercetin probenecid derived from blueberry with small side effect, and a preparation method and application thereof.

In order to solve the problems in the prior art, the application provides the following technical scheme that in the first aspect, the application provides a compound of quercetin probenecid derived from blueberry;

in a second aspect, the present application provides a method for preparing a compound of quercetin probenecid derived from vaccinium bracteatum.

In a third aspect, the application provides a pharmaceutical composition for the treatment of hyperuricemia comprising a compound of quercetin probenecid derived from vaccinium bracteatum.

In a fourth aspect, the application provides an application of a compound of quercetin probenecid derived from blueberry in preparing a pharmaceutical composition for treating hyperuricemia.

The structural formula of the compound of the blueberry-derived quercetin probenecid is shown as a formula (I), wherein the formula is shown as (C ₂₈H₂₇NO₉ S):

the invention relates to a preparation method of a compound of quercetin probenecid from blueberry, which comprises the following steps:

(1) Dissolving the compound 2 probenecid in 1, 4-dioxane, dropwise adding thionyl chloride, and stirring the mixed solution at 95 ℃ for 4-8 hours;

(2) Concentrating the stirred mixed solution in vacuum, dissolving the concentrated solid in a certain amount of 1, 4-dioxane and adding the compound 1 quercetin;

(3) Stirring the mixed solution at 100 ℃ for 12-24 hours, detecting the reaction by TLC, and cooling to room temperature;

(4) Adding triethylamine and water into the mixed solution after stirring, and stirring for 3-5 hours at room temperature;

(5) Concentrating the mixed solution in vacuum after stirring, extracting the concentrated solid with ethyl acetate and water for 2-3 times, separating ethyl acetate and removing excessive water with anhydrous sodium sulfate;

(6) And (3) performing silica gel column chromatography to obtain a compound of the compound 3, namely the quercetin probenecid derived from blueberry.

Further, in the step (1), the molar volume ratio of the compound 2 probenecid to the 1, 4-dioxane is 0.4:10, and the volume ratio of the thionyl chloride to the 1, 4-dioxane is 4.5:10.

Further, in step (2), the molar ratio of quercetin to compound 2 is 1:0.8.

Further, in step (4), the volume ratio of triethylamine to water is 1:1.

Further, in step (5), the volume ratio of ethyl acetate to water is 4:1.

Further, in the step (1), the amount of probenecid compound 2 was 114mg,0.4mmol, the amount of thionyl chloride was 4.5mL, and the amount of 1, 4-dioxane was 10mL.

Further, in the step (2), the amount of quercetin of the compound 1 was 0.5mmol, and in the step (3), the amounts of triethylamine and water were 4mL, respectively.

The invention relates to a pharmaceutical composition for treating hyperuricemia, which comprises a compound of quercetin and probenecid derived from blueberry.

The invention relates to application of a compound of quercetin probenecid derived from blueberry in preparing a pharmaceutical composition for treating hyperuricemia.

Nuclear magnetic hydrogen spectrum of compound 3 ：1H NMR(500MHz,Chloroform-d)δ8.25–8.19(m,2H),7.90–7.83(m,2H),7.46–7.40(m,2H),7.15(s,1H),6.95(s,1H),6.91–6.86(m,2H),6.73(d,J＝2.2Hz,1H),6.48(s,1H),3.97(s,1H),3.03(t,J＝7.4Hz,4H),1.55(p,J＝7.6Hz,4H),0.90(t,J＝7.6Hz,6H).

The quercetin probenecid compound has the advantages of being derived from natural plants, small in side effect, remarkable in treatment effect and the like, and compliance of patients is improved. The compound of the invention has better biological activity and therapeutic effect through structural modification.

Compared with the prior art, the invention has the following advantages that (1) the uric acid reducing performance is that the quercetin derivative shows the capability of reducing serum uric acid level, which is helpful for treating hyperuricemia, preventing gout and other diseases. The compound has remarkable effects of reducing uric acid, blood sugar and serum total cholesterol of experimental mice and improving the serum total protein level.

(3) Anti-inflammatory action this effect may be further enhanced by the combination of quercetin with probenecid due to its anti-inflammatory properties. This may help to alleviate inflammation and pain symptoms caused by hyperuricemia, such as gout. In addition, the antioxidant and anti-inflammatory properties of the quercetin also provide potential application prospects for the compounds in the treatment of cardiovascular diseases, diabetes and other chronic diseases.

(2) Antioxidant properties quercetin is a polyphenol flavonol with antioxidant properties. The structurally modified compound 3 may retain this property and help reduce oxidative stress and protect cells from damage.

(3) Pleiotropic-uric acid is mainly excreted by the kidneys due to the lack of uricase in humans. Compound 3 may reduce serum uric acid levels by promoting uric acid excretion, thereby reducing uric acid crystallization and deposition, preventing gout flares.

(4) Safety through animal experiments, compound 3 showed no significant side effects during administration, showing good safety, which provides important early data support for its use as a drug candidate. The method has repeatability and operability, and is beneficial to realizing the mass production of the compound.

(5) Potential clinical applications since compound 3 shows uric acid, blood glucose and total cholesterol lowering effects in animal models, this suggests that it may have broad clinical application prospects, especially in the treatment of metabolic syndrome and cardiovascular diseases.

Detailed Description

In order to make the technical problems, technical schemes and beneficial effects to be solved more clear, the application is further described in detail below with reference to the embodiments. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the application.

In the present application, the term "and/or" describes an association relationship of an association object, which means that three relationships may exist, for example, a and/or B may mean that a exists alone, a and B exist together, and B exists alone. Wherein A, B may be singular or plural. The character "/" generally indicates that the context-dependent object is an "or" relationship.

In the present application, "at least one" means one or more, and "a plurality" means two or more. "at least one of" or the like means any combination of these items, including any combination of single item(s) or plural items(s). For example, "at least one (a), b, or c)", or "at least one (a, b, and c)", may each represent a, b, c, a-b (i.e., a and b), a-c, b-c, or a-b-c, wherein a, b, c may be single or multiple, respectively.

It should be understood that, in various embodiments of the present application, the sequence number of each process described above does not mean that the execution sequence of some or all of the steps may be executed in parallel or executed sequentially, and the execution sequence of each process should be determined by its functions and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

The terminology used in the embodiments of the application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. As used in this application and the appended claims, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

The weights of the relevant components mentioned in the description of the embodiments of the present application may refer not only to the specific contents of the components, but also to the proportional relationship between the weights of the components, so long as the contents of the relevant components in the description of the embodiments of the present application are scaled up or down within the scope of the disclosure of the embodiments of the present application. Specifically, the mass in the specification of the embodiment of the application can be a mass unit which is known in the chemical industry field such as mu g, mg, g, kg and the like.

The terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated for distinguishing between objects such as substances from each other. For example, a first XX may also be referred to as a second XX, and similarly, a second XX may also be referred to as a first XX, without departing from the scope of embodiments of the application. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include one or more such feature.

Example 1

The first aspect of the invention provides a compound of quercetin probenecid from blueberry, wherein the structural formula of the compound of quercetin probenecid from blueberry is shown as formula (I):

The second aspect of the present invention provides a method for preparing a compound of quercetin probenecid derived from blueberry, comprising the steps of:

(1) Compound 2 is dissolved in 1, 4-dioxane, thionyl chloride is added dropwise, the mixed solution is stirred for 4 hours at 95 ℃, the molar volume ratio of the compound 2 probenecid to the 1, 4-dioxane is 0.4:10, and the volume ratio of the thionyl chloride to the 1, 4-dioxane is 4.5:10.

(2) Concentrating the stirred mixed solution in vacuum, dissolving the concentrated solid in a certain amount of 1, 4-dioxane, and adding quercetin, wherein the molar ratio of quercetin to compound 2 is 1:0.8.

(3) The mixed solution was stirred at 100 ℃ for 19 hours, and cooled to room temperature after completion of the reaction by TLC;

(4) And adding triethylamine and water into the mixed solution after stirring, and stirring for 4 hours at room temperature, wherein the volume ratio of the triethylamine to the water is 1:1.

(5) After stirring, the mixed solution is concentrated in vacuum, the concentrated solid is extracted for 2 to 3 times by ethyl acetate and water, the ethyl acetate is separated and the excessive water is removed by anhydrous sodium sulfate, and the volume ratio of the ethyl acetate to the water is 4:1.

In some embodiments, in step (1), compound 2 probenecid is dissolved in1, 4-dioxane, thionyl chloride is added dropwise, and the mixed solution is stirred at 95 ℃ for 5 hours.

In some embodiments, compound 2 probenecid is dissolved in 1, 4-dioxane, thionyl chloride is added dropwise and the mixed solution is stirred at 95 ℃ for 8 hours.

In some embodiments, in step (3), the mixed solution is stirred at 100 ℃ for 24 hours, and cooled to room temperature after the reaction is detected by TLC.

In some embodiments, in step (3), the mixed solution is stirred at 100 ℃ for 12 hours, and cooled to room temperature after the reaction is completed by TLC.

In some embodiments, in step (4), triethylamine and water are added to the mixed solution after stirring and then stirred at room temperature for 5 hours, wherein the volume ratio of triethylamine to water is 1:1.

In some embodiments, in step (4), triethylamine and water are added to the mixed solution after stirring and then stirred at room temperature for 3 hours, wherein the volume ratio of triethylamine to water is 1:1.

In some embodiments, in step (1), compound 2 probenecid is used in an amount of 114mg,0.4mmol, thionyl chloride is used in an amount of 4.5mL, and 1, 4-dioxane is used in an amount of 10mL.

In some embodiments, compound 1 quercetin is used in an amount of 0.5mmol in step (2), and triethylamine and water are used in an amount of 4mL in step (3), respectively.

A third aspect of the present invention is to provide a pharmaceutical composition for treating hyperuricemia comprising a compound of quercetin probenecid derived from vaccinium bracteatum.

The fourth aspect of the invention is to provide an application of a compound of quercetin probenecid derived from blueberry in preparing a pharmaceutical composition for treating hyperuricemia.

Example 1

(1) Compound 2 probenecid (114 mg,0.4 mmol) was dissolved in 1, 4-dioxane (10 mL), 4.5mL thionyl chloride was added dropwise, the mixed solution was stirred at 95 ℃ for 4 hours, and the stirred mixed solution was concentrated in vacuo;

(2) The concentrated solid was dissolved in 10mL of 1, 4-dioxane and compound 1 quercetin (143 mg,0.5 mmol) was added, the mixed solution was stirred at 100 ℃ for 24 hours, cooled to room temperature after completion of the reaction by TLC, and 4mL of triethylamine and 4mL of water were added to the stirred mixed solution and then stirred at room temperature for 4 hours;

(3) After the completion of stirring, the mixed solution was concentrated in vacuo, the concentrated solid was extracted with ethyl acetate (200 mL) and water (50 mL) 2 to 3 times, ethyl acetate was separated and excess water was removed with anhydrous sodium sulfate, and the compound 3, i.e., a compound obtained from vaccinium bracteatum, quercetin probenecid (183mg, 82%) was obtained after silica gel column chromatography.

Example 2

Example 2 differs from example 1 in that:

in step (1), compound 2 probenecid (114 mg,0.4 mmol) was dissolved in 1, 4-dioxane (10 mL), 4.5mL thionyl chloride was added dropwise, the mixed solution was stirred at 95 ℃ for 8 hours, and the stirred mixed solution was concentrated in vacuo;

In step (2), the concentrated solid was dissolved in 10mL of 1, 4-dioxane and compound 1 quercetin (143 mg,0.5 mmol) was added, the mixed solution was stirred at 100 ℃ for 12 hours, cooled to room temperature after completion of the reaction by TLC, and 4mL of triethylamine and 4mL of water were added to the stirred mixed solution and then stirred at room temperature for 5 hours.

Example 3

Example 3 differs from example 1 in that:

In step (1), compound 2 probenecid (114 mg,0.4 mmol) was dissolved in 1, 4-dioxane (10 mL), 4.5mL thionyl chloride was added dropwise, the mixed solution was stirred at 95 ℃ for 6 hours, and the stirred mixed solution was concentrated in vacuo;

In step (2), the concentrated solid was dissolved in 10mL of 1, 4-dioxane and compound 1 quercetin (143 mg,0.5 mmol) was added, the mixed solution was stirred at 100 ℃ for 20 hours, cooled to room temperature after completion of the reaction by TLC, and 4mL of triethylamine and 4mL of water were added to the stirred mixed solution and then stirred at room temperature for 3 hours.

Test example 1

Test results of blood uric acid of mice

The number of male rats is 60, and each group is divided into 6 groups including positive group (benzbromarone), physiological saline group, CMC model group, compound 3 high-low dosage group and control group.

And (3) preparation of a reagent:

(1) The CMC-Na solution (0.8%) is prepared by weighing 2.4 g CMC-Na, mixing with small amount of purified water to form a lake, adding purified water to 300ml, boiling in beaker to dissolve completely, adding into volumetric flask, and adding purified water to 300ml.

(2) 50Mg/ml hypoxanthine preparation 1g hypoxanthine was weighed and dissolved in 20ml CMC-Na solution.

(3) 0.5Mg/ml of benzbromarone is prepared by weighing 20mg of benzbromarone, and dissolving in 40ml of CMC-Na solution to make the concentration of benzbromarone be 0.5mg/ml.

(4) The quasi-positive medicine compound 3 is prepared by accurately weighing compound 3 (1 g), and preparing 0.5mg/mL, 1.0mg/mL and 2.0mg/mL dosage test solution by using the CMC-Na solution. Each group of animals was dosed once daily (pre-daily weight) for 7 days and each group of animals was dosed intragastrically (i.g.), 2 model group intragastrically (i.g.), and CMC-Na, respectively, with different dosing groups for uric acid levels as indicated in Table 1:

TABLE 1

In the experimental murine model, compound 3 showed the ability to significantly reduce blood uric acid levels. The blood uric acid level of the model group can be obviously reduced by the compound 3 with different doses, and along with the increase of the dose, the uric acid reducing effect is more obvious.

Test example 2

Blood sugar measurement results of laboratory mice

The tests show that the compound 3 with low, medium and high doses has different degrees of effects on the reduction of blood sugar level of mice with hyperuricemia, and the positive control tribromouron group has no obvious effect (table 2). Blood glucose levels for the different dosing groups are shown in table 2:

TABLE 2

Hyperuricemia is associated with a variety of chronic diseases, and compound 3 can reduce blood uric acid levels by regulating blood uric acid levels, possibly contributing to the improvement or prevention of complications such as hypertension, diabetes, metabolic syndrome, kidney and cardiovascular diseases associated with hyperuricemia.

Test example 3

Test mouse Total cholesterol measurement results

Compound 3 was tested at low, medium and high doses with varying degrees of effect on the reduction of serum total cholesterol levels in hyperuricemic mice (table 3). Serum total cholesterol levels for the different dosing groups are shown in table 3:

TABLE 3 Table 3

The experimental results show that compound 3 can positively influence blood glucose and serum total cholesterol levels, showing potential therapeutic effects on metabolic syndrome.

Test example 4

Test mouse Total protein assay results

Through tests, the serum total protein of the hyperuricemia mice is obviously reduced, and the low, medium and high doses of the compound 3 have different degrees of effects on the rise of the serum total protein level of the hyperuricemia mice, and the serum total protein of an experimental group is very close to the normal level of a normal saline group (table 4). Serum total protein levels for the different dosing groups are shown in table 4:

TABLE 4 Table 4

As can be seen from tables 1 to 4, the high dose of compound 3 can significantly reduce uric acid level in blood, and the effect of the low and medium dose of compound 3 on treating hyperuricemia is superior to that of the existing drug tribromoron for treating hyperuricemia, and is superior to that of tribromouron in reducing blood sugar and cholesterol. In conclusion, the compound 3 has a certain treatment effect on hyperuricemia.

The foregoing has shown and described the basic principles, principal features and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the foregoing embodiments, which have been described in the foregoing embodiments and description merely illustrates the principles of the invention, and various changes and modifications may be made therein without departing from the spirit and scope of the invention, the scope of which is defined in the appended claims, specification and their equivalents.

Claims

1. A compound of quercetin probenecid from blueberry is characterized in that the structural formula of the compound of quercetin probenecid from blueberry is shown as the formula (I):

2. the method for preparing a compound of blueberry-derived quercetin probenecid according to claim 1, comprising the steps of:

(2) Concentrating the stirred mixed solution in vacuum, dissolving the concentrated solid in 1, 4-dioxane and adding compound 1 quercetin;

3. The method for producing a compound of quercetin probenecid derived from blueberry according to claim 2, wherein in the step (1), the molar volume ratio of 2 probenecid to 1, 4-dioxane is 0.4:10, and the volume ratio of thionyl chloride to 1, 4-dioxane is 4.5:10.

4. The method for producing a compound of blueberry-derived quercetin probenecid according to claim 2, wherein the molar ratio of quercetin to compound 2 probenecid in step (2) is 1:0.8.

5. The method for producing a compound of quercetin probenecid derived from Oriental blueberry according to claim 2, wherein the volume ratio of triethylamine to water in the step (4) is 1:1.

6. The method for producing a compound of quercetin probenecid derived from Oriental blueberry according to claim 2, wherein the volume ratio of ethyl acetate to water in step (5) is 4:1.

7. The method for producing a compound of blueberry-derived quercetin probenecid according to claim 2, wherein in the step (1), the compound 2 probenecid is used in an amount of 114mg,0.4mmol, thionyl chloride is used in an amount of 4.5mL, and 1, 4-dioxane is used in an amount of 10mL.

8. The method for producing a compound of blueberry-derived quercetin probenecid according to claim 2, wherein in the step (2), the amount of quercetin 1 is 0.5mmol, and in the step (3), the amounts of triethylamine and water are 4mL, respectively.

9. A pharmaceutical composition for the treatment of hyperuricemia, characterized by comprising the compound of blueberry-derived quercetin probenecid according to claim 1.

10. Use of a compound of blueberry-derived quercetin probenecid as claimed in claim 1 for the preparation of a pharmaceutical composition for the treatment of hyperuricemia.