Apigenin derivative and its application in preparing uric acid-lowering drugs

Abstract

The present invention discloses an apigenin derivative, a solvate or pharmaceutically acceptable salt thereof, and its use in the preparation of a uric acid-lowering drug, belonging to the field of pharmaceutical technology. The apigenin derivative of the present invention, having a structure represented by formula (I), is prepared by grafting apigenin onto carnosine. The compound of formula (I) exhibits superior uric acid-lowering effects compared to the raw materials apigenin and carnosine, as well as the traditional positive drug benzbromarone, which also have the effect of lowering blood uric acid.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and more specifically, relates to an apigenin derivative or a solvate or a pharmaceutically acceptable salt thereof and an application thereof in the preparation of a uric acid-lowering drug. Background Art

[0002] Hyperuricemia is a metabolic disease characterized by elevated uric acid levels in the blood. Uric acid is the end product of purine metabolism in the human body. Normal blood uric acid levels are below 360 mmol / L (6 mg / dL) in women and below 400 μmol / L (6.8 mg / dL) in men. The development of hyperuricemia is associated with multiple factors, including lifestyle, nutritional balance, medications, gender, age, and genetics. In particular, nutritional imbalances and increased alcohol consumption contribute to an increased prevalence of the disease among young people. Long-term hyperuricemia can lead to hypertension, hyperlipidemia, hyperglycemia, gouty arthritis, kidney stones, renal failure, and cardiovascular disease. Therefore, there is an urgent need to develop strategies for the treatment and prevention of hyperuricemia. Currently available drugs for the treatment of hyperuricemia, such as allopurinol and benzbromarone, are derived through chemical synthesis. These synthetic drugs often have toxic side effects, whereas drugs derived from natural sources tend to have fewer toxic side effects.

[0003] Apigenin, also known as apigenin, is a naturally occurring flavonoid compound found in a variety of fruits, vegetables, and legumes, with celery containing it at its highest concentration. Existing research has found that apigenin can significantly inhibit the activity of xanthine oxidase, similar to allopurinol in lowering uric acid levels. Currently, a health supplement specifically for gout patients has been developed overseas, using celery seed as its primary ingredient.

[0004] However, the uric acid-lowering effect of apigenin or celery seeds is limited and still needs to be further improved. Summary of the Invention

[0005] 1. Problem to be solved

[0006] In response to the technical problem of further lowering blood uric acid, the purpose of the present invention is to provide an apigenin derivative and its use in lowering uric acid. The compound has a better uric acid-lowering effect than benzbromarone; furthermore, the compound is prepared by the method of apigenin grafted carnosine, both of which are food nutrients and can minimize the side effects of blood uric acid-lowering drugs on the human body.

[0007] 2. Technical Solution

[0008] In order to solve the above problems, the present invention attempts to derivatize apigenin in order to improve its uric acid-lowering effect.

[0009] Carnosine belongs to the histidine dipeptide family, consisting of β-alanine and L-histidine, and is naturally present in vertebrate skeletal muscle and brain tissue. As a food nutrient, carnosine has many biologically active functions, including antioxidant, anti-fatigue, anti-aging, and anti-inflammatory effects. Due to its antioxidant and anti-inflammatory properties, carnosine helps to repair liver and kidney dysfunction in various diseases. For example, carnosine has been found to have a protective effect on hydrogen peroxide-induced oxidative stress in the human kidney and effectively reduce renal damage in diabetic rats. Carnosine's effect on lowering blood uric acid in hyperuricemic rats is achieved by inhibiting inflammation to enhance uric acid excretion and restore liver and kidney dysfunction. Based on this, the present invention uses carnosine grafted with apigenin to achieve the purpose of enhancing apigenin's uric acid-lowering effect.

[0010] The technical solutions adopted in the present invention are as follows:

[0011] The technical solutions of the present invention are as follows:

[0012] The first aspect of the present invention provides an apigenin derivative or a solvate or a pharmaceutically acceptable salt thereof. The apigenin derivative has a structure represented by the following formula (I):

[0013]

[0014] Formula (I).

[0015] The molecular formula of the apigenin derivative is C 26 H 22 N4O7.

[0016] The second aspect of the present invention provides a method for preparing the apigenin derivative or its solvate or pharmaceutically acceptable salt according to the first aspect, comprising the step of reacting apigenin with carnosine.

[0017] As a preferred method for preparing the apigenin derivative or its solvate or pharmaceutically acceptable salt according to the second aspect of the present invention, the preparation method comprises:

[0018] i. a step of reacting apigenin with dimethyl sulfoxide;

[0019] ii. a step of reacting the product of step i with carnosine.

[0020] As a preferred method for preparing the apigenin derivative or its solvate or pharmaceutically acceptable salt according to the second aspect of the present invention, the preparation method further comprises:

[0021] iii. a step of adding triethylamine to the product of step ii.

[0022] The third aspect of the present invention provides use of the apigenin derivative or its solvate or pharmaceutically acceptable salt described in the first aspect in the preparation of a drug for lowering blood uric acid.

[0023] According to any embodiment of the third aspect of the present invention, the drug for lowering blood uric acid comprises:

[0024] Medications that lower blood uric acid by promoting uric acid excretion; or

[0025] Medications that lower blood uric acid by inhibiting uric acid production; or

[0026] Medications that lower serum uric acid by inhibiting uric acid reabsorption; or

[0027] A drug that lowers blood uric acid and is characterized by catalyzing the conversion of uric acid.

[0028] According to the application of any embodiment of the third aspect of the present invention, the drug for lowering blood uric acid is used to treat hyperuricemia.

[0029] The fourth aspect of the present invention provides a pharmaceutical composition comprising the apigenin derivative or a solvate or pharmaceutically acceptable salt thereof according to the first aspect as the sole active ingredient or as one of the active ingredients.

[0030] A fourth aspect of the present invention provides a pharmaceutical composition, which further comprises a pharmaceutically acceptable excipient or carrier.

[0031] According to the pharmaceutical composition of any embodiment of the fourth aspect of the present invention, the amount of the apigenin derivative or its solvate or pharmaceutically acceptable salt contained in the pharmaceutical composition or the dosage to be administered is 10 to 2000 mg.

[0032] Preferably, it is 50 to 1000 mg.

[0033] More preferably, it is 100 to 800 mg.

[0034] The most preferred range is 200-500 mg.

[0035] On this basis, the amount of the apigenin derivative or its solvate or pharmaceutically acceptable salt contained in the pharmaceutical composition or the size of the dose to be administered can be any value within any of the following numerical ranges:

[0036] 10~2000mg, 10~1500mg, 10~1200mg, 10~1000mg, 10~900mg, 10~800mg, 10~700mg, 10~600mg, 10~500mg, 10~400mg, 10~300mg, 10~200mg, 10~100mg;

[0037] 50~2000mg, 50~1500mg, 50~1200mg, 50~1000mg, 50~900mg, 50~800mg, 50~700mg, 50~600mg, 50~500mg, 50~400mg, 50~300mg, 50~200mg, 50~100mg;

[0038] 100~1000mg, 100~900mg, 100~800mg, 100~700mg, 100~600mg, 100~500mg, 100~400mg, 100~300mg, 100~200mg;

[0039] 200~1000mg, 200~900mg, 200~800mg, 200~700mg, 200~600mg, 200~500mg, 200~400mg, 200~300mg;

[0040] 300~1000mg, 300~900mg, 300~800mg, 300~700mg, 300~600mg, 300~500mg, 300~400mg;

[0041] 400~1000mg, 400~900mg, 400~800mg, 400~700mg, 400~600mg, 400~500mg;

[0042] 500~1000mg, 500~900mg, 500~800mg, 500~700mg, 500~600mg;

[0043] 600~1000mg, 600~900mg, 600~800mg, 600~700mg;

[0044] 700~1000mg, 700~900mg, 700~800mg;

[0045] 800~1000mg, 800~900mg;

[0046] 900~1000mg.

[0047] When administered at the above dosages, satisfactory results are achieved, preferably in 1, 2, or 3 doses per day, in whole or in divided doses, or in a sustained-release form. This dosage regimen can be adjusted to provide the optimal therapeutic response. For example, several divided doses may be administered daily, or the dose may be proportionally reduced, depending on the exigencies of the therapeutic situation.

[0048] According to the pharmaceutical composition of any embodiment of the fourth aspect of the present invention, the dose to be administered is a single-dose dose.

[0049] According to the pharmaceutical composition of any embodiment of the fourth aspect of the present invention, the dose to be administered is a daily dose.

[0050] According to any embodiment of the fourth aspect of the present invention, the pharmaceutical composition is administered once, twice or three times a day.

[0051] According to any embodiment of the fourth aspect of the present invention, the pharmaceutical composition is a pharmaceutical composition for administration via the gastrointestinal tract.

[0052] According to any embodiment of the fourth aspect of the present invention, the pharmaceutical composition is a pharmaceutical composition for administration by injection.

[0053] According to any embodiment of the fourth aspect of the present invention, the pharmaceutical composition is a pharmaceutical composition for administration via subcutaneous implantation.

[0054] The pharmaceutical composition of the present invention may be in a variety of dosage forms, any form that allows the active ingredient to effectively reach the mammalian body. For example, it may be selected from: powders, dispersants, tablets, pills, capsules, sustained-release formulations, controlled-release formulations, injections, infusions, and suspensions. Depending on the type of disease to be treated by the composition of the present invention, those skilled in the art can select a dosage form that is convenient for use. From the perspective of ease of preparation and storage, preferred pharmaceutical compositions are solid compositions, particularly tablets and solid-fill or liquid-filled capsules. From the perspective of ease of administration, preferred pharmaceutical compositions are oral formulations. The pharmaceutical composition of the present invention may also be stored in sterilized containers suitable for injection or instillation.

[0055] Pharmaceutical compositions for oral administration include solid dosage forms such as capsules, tablets, dragees, pills, lozenges, powders and granules, which may be prepared with coatings as appropriate.

[0056] Liquid dosage forms for oral administration include solutions, emulsions, suspensions, syrups, and elixirs.

[0057] The formulation and preparation of the pharmaceutical compositions of the present invention in the form of these pharmaceutical preparations can be achieved by methods and experience well known to those skilled in the art.

[0058] According to any embodiment of the fourth aspect of the present invention, the pharmaceutical composition has the following dosage forms: powder, powder, tablet, pill, capsule, sustained-release agent, controlled-release agent, injection, infusion, suspension.

[0059] As pharmaceutical composition, it can be widely used as medicine, functional health care products etc. Specifically, for example tablet (including plain tablet, sugar-coated tablet, film-coated tablet, sublingual tablet, orally disintegrating tablet, buccal tablet etc.), pill, powder, granule, capsule (including soft capsule, microcapsule), buccal tablet, syrup, liquid, emulsion, suspension, controlled release preparation (such as quick-release preparation, sustained-release preparation, sustained-release microcapsule), aerosol, membrane (such as orally disintegrating film, oral mucosa patch), percutaneous absorption type preparation, ointment, lotion, patch, pellet, nasal agent, oral agent or parenteral agent such as pulmonary agent (inhalant) can be enumerated. As long as the pharmaceutical composition of the present invention contains formula (I) active component of the present invention, then can be prepared according to conventional method in the scope that can realize the purpose of the present invention, suitably coordinate commonly used carrier, base and / or additive etc. in preparation field.

[0060] The fifth aspect of the present invention provides a medicine kit, which includes the apigenin derivative or its solvate or pharmaceutically acceptable salt provided by any embodiment of the first aspect of the present invention, and a pharmaceutically acceptable carrier, or includes the pharmaceutical composition of any embodiment of the fourth aspect of the present invention.

[0061] The sixth aspect of the present invention provides use of the apigenin derivative or its solvate or pharmaceutically acceptable salt described in the first aspect in preventing, treating or alleviating hyperuricemia.

[0062] According to any embodiment of the sixth aspect of the present invention, the treatment of hyperuricemia comprises:

[0063] Promote uric acid excretion; or

[0064] inhibiting uric acid production; or

[0065] Inhibit uric acid reabsorption; or

[0066] Catalyzes the conversion of uric acid.

[0067] According to the application of the sixth aspect of the present invention, the apigenin derivative or its solvate or pharmaceutically acceptable salt described in the first aspect is administered as the sole active ingredient of the drug, or the apigenin derivative or its solvate or pharmaceutically acceptable salt described in the first aspect is administered as one of the active ingredients to achieve the therapeutic purpose.

[0068] According to any embodiment of the sixth aspect of the present invention, the amount of the apigenin derivative or its solvate or pharmaceutically acceptable salt or the dosage to be administered is 10 to 2000 mg.

[0069] Preferably, it is 50 to 1000 mg.

[0070] More preferably, it is 100 to 800 mg.

[0071] The most preferred range is 200-500 mg.

[0072] On this basis, the amount of the apigenin derivative or its solvate or pharmaceutically acceptable salt or the size of the dose to be administered can be any value within any of the following numerical ranges:

[0073] 10~2000mg, 10~1500mg, 10~1200mg, 10~1000mg, 10~900mg, 10~800mg, 10~700mg, 10~600mg, 10~500mg, 10~400mg, 10~300mg, 10~200mg, 10~100mg;

[0074] 50~2000mg, 50~1500mg, 50~1200mg, 50~1000mg, 50~900mg, 50~800mg, 50~700mg, 50~600mg, 50~500mg, 50~400mg, 50~300mg, 50~200mg, 50~100mg;

[0075] 100~1000mg, 100~900mg, 100~800mg, 100~700mg, 100~600mg, 100~500mg, 100~400mg, 100~300mg, 100~200mg;

[0076] 200~1000mg, 200~900mg, 200~800mg, 200~700mg, 200~600mg, 200~500mg, 200~400mg, 200~300mg;

[0077] 300~1000mg, 300~900mg, 300~800mg, 300~700mg, 300~600mg, 300~500mg, 300~400mg;

[0078] 400~1000mg, 400~900mg, 400~800mg, 400~700mg, 400~600mg, 400~500mg;

[0079] 500~1000mg, 500~900mg, 500~800mg, 500~700mg, 500~600mg;

[0080] 600~1000mg, 600~900mg, 600~800mg, 600~700mg;

[0081] 700~1000mg, 700~900mg, 700~800mg;

[0082] 800~1000mg, 800~900mg;

[0083] 900~1000mg.

[0084] When administered at the above dosages, satisfactory results are achieved, preferably in 1, 2, or 3 doses per day, in whole or in divided doses, or in a sustained-release form. This dosage regimen can be adjusted to provide the optimal therapeutic response. For example, several divided doses may be administered daily, or the dose may be proportionally reduced, depending on the exigencies of the therapeutic situation.

[0085] According to any embodiment of the sixth aspect of the present invention, the dose to be administered is a single-dose dose.

[0086] According to any embodiment of the sixth aspect of the present invention, the dose to be administered is a daily dose.

[0087] According to any embodiment of the sixth aspect of the present invention, the apigenin derivative or its solvate or pharmaceutically acceptable salt is administered once, twice or three times a day.

[0088] According to the application of any embodiment of the sixth aspect of the present invention, the apigenin derivative or its solvate or pharmaceutically acceptable salt is administered via the gastrointestinal tract.

[0089] According to the application of any embodiment of the sixth aspect of the present invention, the apigenin derivative or its solvate or pharmaceutically acceptable salt is a pharmaceutical composition for administration by injection.

[0090] According to the application of any embodiment of the sixth aspect of the present invention, the apigenin derivative or its solvate or pharmaceutically acceptable salt is a pharmaceutical composition administered via subcutaneous embedding.

[0091] Any embodiment of any aspect of the present invention can be combined with other embodiments without contradiction. In addition, in any embodiment of any aspect of the present invention, any technical feature can be applied to the technical feature in other embodiments without contradiction.

[0092] Under the premise that there is no contradiction, any technical feature possessed by any aspect of the present invention or any embodiment of any aspect is also applicable to any other embodiment or any embodiment of any other aspect. Of course, when applicable to each other, the corresponding features may be appropriately modified if necessary.

[0093] The term "solvate" used herein refers to a compound that carries solvent molecules, for example, the solvate may be a hydrate.

[0094] In the present invention, the term "comprising" or "containing" indicates that the active ingredient can be used together with other components in the pharmaceutical composition of the present invention.

[0095] The compositions of the present invention also include isomers, solvates, precursors, or pharmaceutically acceptable salts thereof of the specific compounds in the pharmaceutical composition, as long as they have the same or substantially the same function as the specific compounds in the composition.

[0096] In the present invention, a "pharmaceutically acceptable" ingredient is a substance that is suitable for use in humans and / or animals without excessive adverse side effects (such as toxicity, irritation, and allergic reactions), that is, with a reasonable benefit / risk ratio.

[0097] In the present invention, "pharmaceutically acceptable salts" refer to acid addition salts of a compound with a non-toxic acid, including salts formed by reactions with inorganic acids, organic acids, alkali metals or alkaline earth metals. These salts include, but are not limited to:

[0098] (1) Salts formed with the following inorganic acids: hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid;

[0099] (2) Salts formed with organic acids such as acetic acid, oxalic acid, succinic acid, tartaric acid, methanesulfonic acid, maleic acid, or arginine. Other salts include salts formed with alkali metals or alkaline earth metals (such as sodium, potassium, calcium, or magnesium) in the form of esters, carbamates, or other conventional "prodrugs."

[0100] The compounds in the compositions of the present invention have one or more asymmetric centers and may therefore exist as racemic mixtures, individual enantiomers, individual diastereomers, diastereomeric mixtures, cis- or trans-isomers.

[0101] In the present invention, a "pharmaceutically acceptable carrier" is a pharmaceutically acceptable solvent, suspending agent, or excipient used to deliver the composition, isomer, solvate, or pharmaceutically acceptable salt thereof of the present invention to animals or humans. The carrier can be a liquid or a solid.

[0102] The "pharmaceutically acceptable carriers or excipients" in the present invention include but are not limited to preservatives, wetting agents, emulsifiers and dispersants.

[0103] The pharmaceutical composition of the present invention can be used as the sole drug, or can be used in combination with one or more other drugs that have synergistic and / or synergistic effects with the substance of the present invention on physiological activity. Combination therapy can be achieved by administering the individual therapeutic components simultaneously, sequentially or separately.

[0104] The actual dosage level and route of administration of the active ingredients in the pharmaceutical compositions of the present invention can be varied to obtain an amount of active substance effective to achieve the desired therapeutic response in a particular patient. The dosage level should be selected based on the activity of the specific active substance, the route of administration, the severity of the condition being treated, and the condition and medical history of the patient being treated. However, it is common practice in the art to start the dosage of the active substance at a level below that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0105] When used for the above-mentioned treatment and / or prevention of the present invention or other treatment and / or prevention, a pharmaceutical composition of the present invention in a therapeutic and / or preventive effective amount can be used alone. Alternatively, the pharmaceutical composition of the present invention can be administered in combination with one or more pharmaceutically acceptable carriers or excipients.

[0106] As used herein, the term "treat" refers to the management and care of a patient for the purpose of combating a condition (e.g., a disease or disorder). The term is intended to encompass the full range of treatments for a given condition suffered by a patient, such as administering a composition to alleviate symptoms or complications, to delay progression of the condition, and / or to cure or eliminate the condition. The patient to be treated is preferably a mammal, particularly a human.

[0107] As used herein, the term "therapeutically and / or prophylactically effective amount" of a pharmaceutical composition of the present invention refers to an amount sufficient to cure, alleviate, or partially arrest the clinical manifestations of a given disease and its complications in a therapeutic intervention comprising administration of the pharmaceutical composition. An amount sufficient to achieve the above is defined as a "therapeutically and / or prophylactically effective amount." The effective amount for each purpose will depend on the severity of the disease or injury, as well as the weight and general condition of the subject. However, it should be understood that the total daily dosage of the pharmaceutical composition of the present invention should be determined by the attending physician within the scope of sound medical judgment. For any particular patient, the specific therapeutically effective dosage level will depend on a variety of factors, including the disorder being treated and the severity of that disorder; the activity of the specific pharmaceutical composition employed; the specific pharmaceutical composition employed; the patient's age, weight, general health, sex, and diet; the time of administration, route of administration, and excretion rate of the specific pharmaceutical composition employed; the duration of treatment; other drugs used in combination or concurrently with the pharmaceutical composition employed; and similar factors well known in the medical field. For example, it is common practice in the art to start the dosage of the pharmaceutical composition at a level below that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.

[0108] The compound (active ingredient) in the pharmaceutical composition used in the present invention can be administered alone as a pure compound in a single or multiple doses, or in combination with a pharmaceutically acceptable carrier or excipient. The pharmaceutical composition according to the present invention can be formulated using a pharmaceutically acceptable carrier or excipient and any other known adjuvants and vehicles according to conventional techniques.

[0109] 3. Beneficial effects

[0110] Compared to existing technologies, the present invention provides a novel compound with enhanced serum uric acid-lowering effects. This compound exhibits superior serum uric acid-lowering effects compared to apigenin, carnosine, and the traditional positive-acting drug benzbromarone, which also have similar effects. Furthermore, apigenin and carnosine both possess excellent biocompatibility, significantly increasing the market potential of the present apigenin derivatives and potentially contributing to the development of uric acid-lowering drugs. DETAILED DESCRIPTION

[0111] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0112] If the specific conditions are not specified in the examples, the experiments were carried out under conventional conditions or those recommended by the manufacturer. All reagents or instruments used, if the manufacturer is not specified, are commercially available conventional products.

[0113] Concentration, amount and other numerical data can be presented in range format herein.It should be understood that such range format is used only for convenience and brevity, and should be flexibly interpreted as not only including the numerical value clearly stated as the range limit, but also including all individual numerical values ​​or sub-ranges encompassed within the range, just as each numerical value and sub-range is clearly stated.For example, a numerical range of about 1 to about 4.5 should be interpreted as including not only the clearly stated limit value of 1 to about 4.5, but also including individual numbers (such as 2, 3, 4) and sub-ranges (such as 1 to 3, 2 to 4, etc.).The same principle applies to the range of only narrating a numerical value, such as "less than about 4.5", which should be interpreted as including all the above-mentioned values ​​and ranges.In addition, no matter how the breadth of the described range or feature is, this interpretation should be applied.

[0114] The present invention will be further described below with reference to specific embodiments.

[0115] Example 1

[0116] Preparation of the compound of formula (I) (Compound 3) (Quercetin-3-O-glucuronide coupled with (1-benzyl-4-piperidinyl)methylamine compound)

[0117]

[0118] Compound 3

[0119]

[0120] Apigenin (270 mg, 1 mmol) was dissolved in 1,4-dioxane (25 mL), and thionyl chloride (9.4 mL, 0.13 mol) was added dropwise. The resulting mixture was stirred at 70°C for 4 hours and then concentrated in vacuo. The concentrated solid was added to 1,4-dioxane (25 mL) and a predetermined amount of carnosine (272 mg, 1.2 mmol). The mixture was stirred at 70°C for 2 hours. Triethylamine (8.3 mL), ethanol (4.1 mL), and water (8.3 mL) were added to the stirred mixture, and the mixture was concentrated again. The concentrated solid was extracted two or three times with diethyl ether (50 mL) and water (15 mL). The diethyl ether was separated, and excess water was removed with anhydrous sodium sulfate. The product was concentrated in vacuo and purified by silica gel column chromatography (dichloromethane:methanol = 100:1) to obtain compound (I) (434 mg, 0.91 mmol, 91%).

[0121] H NMR spectrum of compound 3: 1 H NMR (500 MHz, Chloroform-d) δ 8.50 (s, 1H), δ8.90 (s, 1H), δ 7.97 (d, J = 9.5 Hz, 1H), 7.91 - 7.85 (m, 2H), 7.53 (dd, J =5.9, 1.6 Hz, 1H), 7.25 – 7.19 (m, 2H), 6.82 (dd, J = 4.8, 1.6 Hz, 1H), 6.75(s, 1H), 6.52 (d, J = 1.8 Hz, 1H), 6.25 (d, J = 1.8 Hz, 1H), 4.68 (dt, J =9.7, 6.7 Hz, 1H), 4.23 (dd, J = 5.9, 4.9 Hz, 1H), 3.35 (t, J = 6.6 Hz, 2H), 3.09 (dd, J = 14.6, 6.8 Hz, 1H), 3.03 (dd, J = 14.7, 6.8 Hz, 1H), 2.87 (ttd,J = 6.4, 4.9, 1.6 Hz, 2H), 2.35 (d, J = 9.7 Hz, 2H).

[0122] Example 2

[0123] Effects of compound 3 on blood uric acid in experimental mice

[0124] Sixty male mice were randomly divided into six groups (10 mice in each group): a positive group (benzbromarone), a normal saline group, a CMC model group, high, medium and low dose groups of compound 3, and a raw material (apigenin and carnosine) control group.

[0125] Reagent preparation:

[0126] (1) Preparation of CMC-Na solution (0.8%): Weigh 2.4 g of CMC-Na, first mix it into a lake with a small amount of pure water, then dilute it to 300 ml with pure water, pour it into a beaker and boil it to completely dissolve it, then pour it into a volumetric flask and make up to 300 ml with pure water.

[0127] (2) Preparation of 50 mg / ml hypoxanthine: Weigh 1 g hypoxanthine and dissolve it in 20 ml CMC-Na solution.

[0128] (3) Preparation of 0.5 mg / ml benzbromarone: Weigh 20 mg of benzbromarone and dissolve it in 40 ml of CMC-Na solution to a concentration of 0.5 mg / ml.

[0129] (4) Preparation of 1.0 mg / ml raw material control group: Weigh 40 mg of apigenin and 40 mg of carnosine, dissolve in 40 ml of CMC-Na solution to a concentration of 1.0 mg / ml.

[0130] (5) Preparation of the pseudo-positive drug compound 3: Compound 3 (1 g) was accurately weighed and prepared into 0.5 mg / mL, 1.0 mg / mL, and 2.0 mg / mL dose test solutions using the above CMC-Na solution. Each group of test animals was given the drug once a day (weighed before each day of administration) for 7 consecutive days. The experimental method for each animal group was as follows: animals in groups 1, 3, 4, 5, 6, and 7 were given intragastrically (ig), and CMC-Na was given intragastrically (ig) to group 2 of the model group. The specific conditions are shown in Table 1:

[0131] Table 1 Groups, dosing methods and dosing volumes

[0132]

[0133] Thirty minutes after the last administration on day 8, all groups except the saline group received an intraperitoneal (ip) injection of 50 mg / ml hypoxanthine at a dose of 1000 mg / kg body weight. 30-40 minutes after administration, the eyeballs were immediately removed and blood was collected from the eyes. The blood was placed in a centrifuge tube and centrifuged at room temperature (15,000 rpm, 4 minutes) until the serum seeped out. The upper layer of serum was aspirated and placed in a capped sample tube. Based on the volume of the serum obtained, the volume was adjusted to the required volume with saline, and the blood uric acid concentration was measured on a Beckman LX20 fully automatic biochemical analyzer.

[0134] The test showed that compound 3 had different degrees of effect on reducing the serum uric acid level in hyperuricemia mice, and within the scope of this experimental design, the effect of the positive control benzbromarone group was not significant (Table 2).

[0135] Table 2 Effects of test samples on blood uric acid in experimental mice (μM)

[0136]

[0137] Example 3

[0138] Determination of the body weight of experimental mice in Example 2

[0139] After testing, the weight of each group in Example 2 (normal saline group, CMC model group, low-dose compound 3 group, medium-dose compound 3 group, high-dose compound 3 group, and benzbromarone group) increased after 7 days of feeding, and within the scope of this experimental design, the level of weight gain of the animals in each group was not much different (Table 3).

[0140] Table 3 Effects of test drugs on the body weight of experimental mice (g)

[0141]

[0142] Note: There was no significant difference in weight gain among the groups.

[0143] Example 4

[0144] Acute toxicity test in rats

[0145] Acute toxicity experiments were conducted on 10 rats (~240 g / rat) in each of the compound 3 and blank groups. The experimental group was given compound 3 (50 times the dose of the medium-dose group) by oral gavage every day (concentration 50 mg / ml, dosing volume 0.1 ml / 10 g, once a day). The blank group was fed normally and no sample was given.

[0146] After 7 consecutive days of feeding, no significant difference was found in the body weight of rats in the experimental group and the blank group, which indicates that compound 3 is relatively safe.

[0147] The above description is merely an illustrative description of the present invention and its embodiments, which is not restrictive. The embodiment shown in the embodiment is only one embodiment of the present invention, and the actual embodiment is not limited thereto. Therefore, if a person skilled in the art is inspired by the above description and, without departing from the purpose of the present invention, designs an embodiment and examples similar to the technical solution without creatively designing, they shall fall within the scope of protection of the present invention.

Claims

1. An apigenin derivative or a pharmaceutically acceptable salt thereof, characterized in that: Apigenin derivatives have the structure shown in the following formula (I): Formula (I).

2. The method for preparing the apigenin derivative or pharmaceutically acceptable salt according to claim 1, characterized in that: The method comprises the steps of reacting apigenin with carnosine; specifically comprising the following steps: Apigenin was dissolved in 1,4-dioxane, thionyl chloride was added dropwise, and the resulting mixed solution was stirred at 70°C for 4 hours, and the stirred mixed solution was concentrated in vacuo; The concentrated solid was added to 1,4-dioxane and carnosine was added, and the mixed solution was stirred at 70°C for 2 hours; Add triethylamine, ethanol and water to the mixed solution after stirring, and then concentrate the mixed solution again; The concentrated solid was extracted two to three times with diethyl ether and water, the diethyl ether was separated and excess water was removed with anhydrous sodium sulfate, the solid was concentrated in vacuo, and the solid was purified by silica gel column chromatography with dichloromethane:methanol=100:1 to obtain the compound of formula (I).

3. The method for preparing the apigenin derivative or pharmaceutically acceptable salt according to claim 2, characterized in that: The content of the apigenin is 1 mmol, and the content of the carnosine is 1.2 mmol.

4. Use of the apigenin derivative or pharmaceutically acceptable salt according to claim 1 in the preparation of a drug for lowering blood uric acid.

5. The use according to claim 4, characterized in that The blood uric acid lowering drug is: Medications that lower blood uric acid by promoting uric acid excretion; or Medications that lower blood uric acid by inhibiting uric acid production; or Medications that lower serum uric acid by inhibiting uric acid reabsorption; or A drug that lowers blood uric acid and is characterized by catalyzing the conversion of uric acid.

6. The use according to claim 4, characterized in that The blood uric acid lowering drug is used to prevent, treat or alleviate hyperuricemia.

7. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises the apigenin derivative or pharmaceutically acceptable salt according to claim 1 as the sole active ingredient or as one of the active ingredients.

8. The pharmaceutical composition according to claim 7, further comprising a pharmaceutically acceptable excipient or carrier.

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