RU2784896C2 - Medical use of anemoside b4 against acute gouty arthritis - Google Patents

Abstract

FIELD: medicine; pharmaceutics.

SUBSTANCE: group of inventions relates to the field of medicine and pharmaceutics; it can be used for the treatment of acute gouty arthritis. The use of anemoside B4 for the production of a drug for the treatment of acute gouty arthritis, as well as a method for the treatment of acute gouty arthritis, including a stage of administration of anemoside B4 to a patient who needs it, are proposed.

EFFECT: invention provides a new alternative strategy of treatment of acute gouty arthritis.

13 cl, 2 tbl, 2 ex

Description

Cross-references to related applications

The present application claims priority under Application No. 201811214694.3, filed October 18, 2018 and entitled "Medical Use of Anemoside B4 Against Acute Gouty Arthritis", the contents of which are hereby incorporated by reference in their entirety.

Technical field

The present invention generally relates to the field of medicine, and in particular relates to a new medical use of anemoside B4.

State of the art

The term gout encompasses a group of diseases in which long-term disturbances in purine metabolism and elevated levels of uric acid in the blood cause tissue damage. Acute gouty arthritis is a typical symptom of gout as a characteristic acute inflammatory reaction caused by the deposition of urate crystals in the joints and periarticular tissues (especially in the ankle and foot). Attacks often occur during the night, and the patient may wake up in pain and then be unable to sleep throughout the night. Acute gouty arthritis often recurs and may progress to polyarthritis or migratory arthritis over time. The affected joints are red, swollen, hot and painful to palpation, their mobility is limited, which seriously affects the quality of life of patients. In China, in order to continuously improve the living standards of the population, due to the increase in the consumption of foods high in purines, proteins and fats, not only the incidence of gout has increased rapidly, but the disease itself has gradually “rejuvenated”, and now there are even patients of adolescence. Over the next 10 years, gout is predicted to become the second most common metabolic disorder in China, behind only diabetes.

Currently commonly used drugs for the treatment of acute gouty arthritis are non-steroidal anti-inflammatory drugs (such as diclofenac sodium, etc.), colchicine and glucocorticoids. Although these drugs achieve some clinical results, associated side effects and adverse reactions are inevitable, such as gastrointestinal reactions to traditional non-steroidal anti-inflammatory drugs; adverse reactions to the use of COX-2 inhibitors in the cardiovascular system; bone marrow suppression, liver damage, allergic reactions and colchicine neurotoxicity; rebound effect of glucocorticoid withdrawal, etc. Therefore, one of the most active areas of research in the field of medicine has become the search for new drugs for the treatment of acute gouty arthritis.

The traditional Chinese medicine Radix Pulsatillas is the dried root of Pulsatilla chinensis (Bge.) Regel, a plant from the genus Pulsatilla of the Ranunculaceae family, and was first described in the classic pharmacological work on herbs and medicines, Shen Nong's Canon of Roots and Herbs. ". Its nature and taste are bitter and cold, and it has the ability to eliminate heat and detoxify, to cool the blood to stop diarrhea, to drain moisture and kill insects, etc. It is used to neutralize heat-labile toxins, treat dysentery with bloody diarrhea, tropical malaria, intermittent fever, epistaxis and hemorrhoids. In the course of modern pharmacological research, Radix Pulsatillae has been found to have more diverse activities, including broad spectrum bactericidal, antitumor, anti-inflammatory, immune function enhancement, and so on.

Radix Pulsatillae is rich in triterpene saponins. Anemoside B4 refers to pentacyclic triterpene lupine-type saponins with the structural formula shown in FIG. one.

Anemoside B4 is characterized by strong action - in particular, in the Chinese patent application for invention with the publication number CN 105213410 A (publication date January 6, 2016), the use of anemoside B4 as an immunomodulator in drugs for the treatment of acute inflammation is disclosed. Inflammation includes acute kidney injury, acute liver injury, and acute lung injury caused by overexpression of inflammatory factors. Another example is Chinese patent application CN 105535004 A (published on May 4, 2016), which discloses the use of these compounds as an inhibitor of EV71 virus for the preparation of drugs against viral pemphigus of the mouth and extremities. However, until now there have been no reports of the use of anemoside B4 for the treatment of acute gouty arthritis.

The essence of the invention

In an effort to make up for the shortcomings of the prior art, the present invention provides a novel medical use for anemoside B4 in the treatment of acute gouty arthritis.

To achieve the above technical results, the present invention includes the following technical solutions:

The use of anemoside B4 for the production of drugs for the treatment of acute gouty arthritis.

As a preferred embodiment of the present invention, the use of anemoside B4 as the sole active pharmaceutical ingredient for the preparation of medicaments for the treatment of acute gouty arthritis is proposed.

As another preferred embodiment, the present invention provides the use of anemoside B4 and other active pharmaceutical ingredients for the preparation of medicaments for the treatment of acute gouty arthritis, wherein the other active pharmaceutical ingredients are selected from one or more of a non-steroidal anti-inflammatory drug, colchicine, and a glucocorticoid.

Said non-steroidal anti-inflammatory drug includes, without limitation, ibuprofen, indomethacin, nimesulide, naproxen, nabumetone, sodium diclofenac, sodium loxoprofen, rofecoxib, celecoxib, etoricoxib, and the like.

Said glucocorticoid includes, without limitation, prednisone, meprednisone, betamethasone, beclomethasone dipropionate, diprospan, prednisolone, hydrocortisone, and dexamethasone.

Preferably, the medicinal product also contains pharmaceutically acceptable excipients.

Preferably, the drug is selected from one or more oral drugs and parenteral drugs.

Preferably, said parenteral drug is selected from one or more injectable drugs, rectal drugs and inhalation (pulmonary) drugs, more preferably selected from one or more injectable drugs and rectal drugs.

Said injectable drug is selected from one or more subcutaneous injection drugs, intramuscular injectable drugs, and intravenous infusion drugs; and

The drug for rectal administration is selected from rectal suppositories and/or rectal infusion drugs.

Pharmaceutically acceptable excipients of the present invention include, without limitation (1) diluents such as starch, powdered sugar, dextrin, lactose, pregelatinized starch, microcrystalline fibers, inorganic calcium salts (such as calcium sulfate, calcium hydrogen phosphate, medicinal calcium carbonate, etc. .d.), mannitol, vegetable oil, polyethylene glycol, cocoa butter, semi-synthetic or fully synthetic fatty acid glycerides, glycerogelatin, etc.; (2) binders such as distilled water, ethanol, starch suspension, povidone, sodium carboxymethylcellulose, hydroxypropylcellulose, methylcellulose and ethylcellulose, hypromellose, etc.; (3) disintegrators such as dry starch, sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, croscarmellose sodium, crospovidone, etc.; (4) lubricants such as magnesium stearate, fine silica gel, talc, hydrogenated vegetable oil, polyethylene glycols, magnesium lauryl sulfate, etc.; (5) solvents such as water for injection, ethanol, etc.; (6) preservatives such as benzoic acid and its salts, sorbic acid and its salts, parabens, etc.

Preferably, the suitable subject is a mammal, and more preferably a human.

For human subjects, the amount of anemoside B4 to administer is typically 0.4-1.6 mg/kg body weight per day for adults (weighing 70 kg), more preferably 0.4-1.6 mg/kg body weight per day. a total of one or more times a day.

The present invention further provides a method for treating acute gouty arthritis which comprises the step of administering anemoside B4 to a patient in need.

Preferably, the method used to treat acute gouty arthritis comprises the step of administering anemoside B4 to a patient in need by subcutaneous injection, intramuscular injection or intravenous infusion.

Preferably, the patient in need is a mammal, more preferably a human.

Preferably, the method of treating acute gouty arthritis comprises the step of administering anemoside B4 at a dose of 0.4-1.6 mg/kg of body weight to a person in need once or several times a day.

As a preferred embodiment of the present invention, a method for the treatment of acute gouty arthritis is provided, which includes the step of administering anemoside B4 at a dose of 0.4-1.6 mg/kg of body weight to a patient in need once or several times a day by subcutaneous injection, intramuscular injection or intravenous injection .

The present inventors have proven through numerous animal experiments that intravenous, subcutaneous or intramuscular injection of anemoside B4 can significantly reduce joint swelling caused by acute gouty arthritis. Therefore, anemoside B4 injection is expected to provide a novel, alternative treatment strategy for acute gouty arthritis.

Best Mode for Carrying out the Invention

Hereinafter, the present invention will be illustrated by specific examples. Those skilled in the art will appreciate that these examples are provided for the purpose of illustrating the present invention only and do not in any way limit the scope of protection of the present invention.

All experimental procedures in the following examples, unless otherwise noted, are conventional methods. All starting materials, reagents, materials, etc. used in the following examples are commercially available products, unless otherwise noted.

Example 1 Preliminary study of the effect of anemoside B4 on the course of acute gouty arthritis induced by sodium urate in rats

1. Experimental materials

1.1 Investigational medicinal products: Anemoside B4 injection drug (obtained for internal use, hereinafter referred to as "B4 injection solution"); colchicine, batch number: 171116, Xishuangbanna Banna Pharmaceutical Co., Ltd.

The above injection solution B4 was prepared in the following way:

A predetermined amount of anemoside B4 raw material was accurately weighed, to which then a suitable amount of water for injection was added, and then anemoside B4 raw material was completely dissolved for stirring with a magnetic stirrer, activated carbon was added to the solution in an amount of 0.10% by weight of the solution. The mixture was heated in a water bath to a temperature of 100°C and stirred for 15 minutes, then diluted to 100 ml with water for injection. The resulting solution was shaken and filtered through a 0.22 μm microporous membrane to remove activated charcoal. Accurately measured 2 ml of the intermediate liquid and transferred into a 5 ml ampoule, followed by sterilization at 115°C for 30 minutes to obtain an injection solution B4.

1.2 Reagent: sodium urate, Sigma company; SKU: U2875-5G; batch number: BCBS7438

1.3 Animals: 60 Sprague-Dawley male rats, 180~220 g, purchased from Hunan SJA Laboratory Animal Co., Ltd., were used for experiments one week after adaptive feeding.

1.4 Measuring instrument: Toe volume measuring instrument, Model: YLS-7B, Huaibei Zhenghua Biological Instrument Equipment Co., Ltd.

2. Experimental technique

2.1 Preparation of crystals and suspension of (single) sodium urate: 5 ml of 1 mol/l NaOH solution and 800 mg of sodium urate were added to 155 ml of depyrogenated sterilized water for injection and heated to boiling. The sodium urate was completely dissolved and then cooled naturally to stir. After that, HCl 1 mol/l was instilled until a pH of 7.0 was reached, for this the solution became milky white, and immediately centrifuged at 3000 rpm for 2 minutes. The crystals were collected, dried in a dry-air thermostat at 60°C, placed in an Eppendorf-type tube, and stored at 4°C. Prior to use, monosodium urate crystals were autoclaved at 121°C for 30 minutes, and dispersed in phosphate-buffered saline until the desired concentration was obtained, i.e. obtaining a suspension of monosubstituted sodium urate. A smear preparation of the suspension was performed, and long spindle-shaped crystals were observed under an optical microscope.

2.2 Grouping, drug administration and modeling: 60 rats were randomly divided into 6 groups according to body weight, 10 rats in each group: (1) reference group; (2) colchicine group (0.3 mg/kg); (3) solution group B4 for subcutaneous injection (5 mg/kg*2); (4) solution group B4 for intravenous injection (5 mg/kg*2); (5) solution group B4 for intramuscular injections at high doses (5 mg/kg*2); (6) solution group B4 for intramuscular injection at low doses (2.5 mg/kg*2); normal group (5 rats). The normal group and the reference group were injected with the same volume of saline, and each of the B4 groups was given the respective drugs by the respective administration for 3 days, twice a day, with an interval of 4 hours between two injections.

Method of building the model: on the day of modeling, the model was started to build 1 hour after the administration of drugs in drug groups. Including:

(1) Reference group: each rat was injected with 40 mg/ml suspension of monosodium urate in the back of the right ankle at a dose of 0.2 ml/rat;

(2) Colchicine group: The colchicine group was administered the drug once on the simulation day, and 1 hour after administration, each rat was injected with 40mg/ml suspension of monosodium urate from the dorsal side of the right ankle at a dose of 0.2ml/rat;

(3)~(6) Group B4: 1 hour after the last injection, each rat was injected with 40mg/ml suspension of monosodium urate from the dorsal side of the right ankle joint at a dose of 0.2ml/rat, and 3 hours after the simulation of each experimental group was additionally administered the drug once.

2.3 Measurement of joint swelling: before modeling and 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, 24 hours after modeling, the volume of the right ankle joint was measured, and the degree of swelling by the combined volume was calculated from the ratio of the volume of the joint after modeling and the volume of the joint before modeling.

3. Experimental results

See Table 1.

The data shown in Table 1 showed that, compared with the normal group, the joints of rats in the reference group swelled significantly 2 hours after simulation, the swelling peaked at 8 hours~12 hours after simulation, and spontaneously decreased in volume 24 hours after simulation. modeling. Compared with the reference group, the degree of swelling of the joints in each group with the introduction of drugs decreased to a different extent. Including, 2~6 hours after simulation in the B4 group for subcutaneous injection, 2~4 hours after simulation in the B4 group for intramuscular injection, and 4~6 hours after simulation in the B4 group for intravenous injection, and after 4 hours after simulation in the colchicine group, the degree of joint swelling was significantly reduced with significant differences. No significant difference was observed between the B4 groups and the colchicine group, indicating that for the use of the appropriate route of administration and dosage in the experiment, the effect of B4 on reducing acute joint swelling in rats induced by monosodium urate crystals was equivalent to that of colchicine.

4. Conclusion on the results of the experiment

The results of this experiment showed that B4 is able to significantly relieve joint swelling in acute gouty arthritis by subcutaneous injections, intramuscular injections and intravenous injections.

Example 2 Additional study of the effect of anemoside B4 on the course of acute gouty arthritis induced by sodium urate in rats

Based on Example 1, the dosage of anemoside B4 was reduced, and the effect on relieving joint swelling in rats with sodium urate-induced acute gouty arthritis was further investigated.

1. Experimental materials: the same as in section 1 in Example 1.

2. Experimental technique

2.1 Preparation of crystals and suspension of dibasic sodium urate: same as in section 2.1 in Example 1.

2.2. Grouping, introduction, and model creation:

2.2.1 Grouping: 50 rats were randomly divided into 5 groups according to body weight, 10 rats in each group:

(1) reference group; (2) colchicine group (0.15 mg/kg); (3) group B4 for subcutaneous injection (2.5 mg/kg*2); (4) group B4 for intramuscular injections (2.5 mg/kg*2); (5) group B4 for intravenous injection (2.5 mg/kg*2).

2.2.2 Administration: The reference group was injected with the same volume of saline, and each treatment group (including the B4 groups and the colchicine group) was injected with the drug only on the day of model creation. Including the colchicine group, the drug was administered once, and each B4 group was sequentially injected twice with an interval of 4 hours.

2.2.3 Model creation and measurement of joint swelling: The model was started 1 hour after administration of the drug in drug groups, and the procedure was the same as in Example 1. Before modeling and after 2 hours, 4 hours, 6 hours, 8 hours and 12 hours after modeling, the volume of the right ankle joint was measured, and the degree of swelling was calculated from the ratio of the volume of the joint after modeling and the volume of the joint before modeling.

3. Results of experiments:

See Table 2.

The data shown in Table 2 showed that the degree of swelling in the reference group peaked at 8h~12h after simulation. Compared with the reference group, the degree of joint swelling in each drug group decreased to a different extent, in particular, 6~12 hours after simulation in the B4 group for subcutaneous injection, 8~12 hours after simulation in the B4 group for intramuscular injection, and after 8 ~12 hours after simulation in the B4 group for intravenous injection, and 6~12 hours after simulation in the colchicine group, the degree of joint swelling obviously decreased and was significantly different compared to the reference group. No significant difference was observed between the B4 groups and the colchicine group, indicating that, using the appropriate route of administration and dosage in the experiment, the effect of B4 on reducing joint swelling in sodium urate-induced acute gouty arthritis in rats was equivalent to that of colchicine. Although no significant difference was observed, the degree of joint swelling in the B4 subcutaneous injection group was the smallest in all groups 2~8 hours after simulation, and the degree of joint swelling in the B4 intramuscular injection group was the smallest in all groups 12 hours after simulation, which indicates a slightly higher efficiency.

4. Conclusion on the results of the experiment

The results of this experiment showed that, similar to the results of Example 1, B4 injection has a significant effect on relieving joint swelling in acute gouty arthritis.

Claims (13)

1. Use of anemoside B4 for the preparation of a medicament for the treatment of acute gouty arthritis. 2. The use according to claim 1, characterized by the use of anemoside B4 as the sole active pharmaceutical ingredient for the preparation of a medicament for the treatment of acute gouty arthritis. 3. The use according to claim 1, characterized by the use of anemoside B4 and other active pharmaceutical ingredients for the preparation of a medicament for the treatment of acute gouty arthritis, in which the other active pharmaceutical ingredients are selected from one or more non-steroidal anti-inflammatory drugs, colchicine and glucocorticoid. 4. Use according to claim 3, characterized in that the non-steroidal anti-inflammatory drug is selected from the group consisting of one or more of the following agents: ibuprofen, indomethacin, nimesulide, naproxen, nabumeton, diclofenac sodium, loxoprofen sodium, rofecoxib, celecoxib and etoricoxib. 5. Use according to claim 3, characterized in that the glucocorticoid is selected from one or more of the following: prednisone, meprednisone, betamethasone, beclomethasone dipropionate, diprospan, prednisolone, hydrocortisone and dexamethasone. 6. Use according to any one of claims 1 to 5, characterized in that the medicinal product additionally contains pharmaceutically acceptable excipients. 7. Use according to any one of claims 1 to 6, characterized in that the drug is selected from one or more oral drugs and parenteral drugs. 8. Use according to claim 7, characterized in that the parenteral drug is selected from one or more injectable drugs, rectal drugs and inhalation drugs and is preferably selected from one or more injectable drugs and rectal drugs. introductions. 9. Use according to claim 8, characterized in that the injectable drug is selected from one or more of the following drugs: drugs for subcutaneous injection, drugs for intramuscular injection and drugs for intravenous infusion; and the drug for rectal administration is selected from rectal suppositories and/or rectal infusion drugs. 10. Use according to any one of claims 1 to 9, characterized in that the subject is a mammal, preferably a human. 11. A method for the treatment of acute gouty arthritis, comprising the step of administering anemoside B4 to a patient in need at a dosage of 2.5 or 5.0 mg/kg of body weight twice a day. 12. The method according to claim 11, characterized in that the method includes the step of administering anemoside B4 to a patient in need by subcutaneous injection, intramuscular injection or intravenous infusion. 13. The method according to claim 11 or 12, characterized in that the patient in need is a mammal, preferably a human.