Medicament for dissolving tophus and application thereof

Abstract

The invention discloses a medicament for dissolving tophus and application of the medicament, and belongs to the technical field of tophus dissolution. The medicament comprises potassium carbonate, and the concentration of the potassium carbonate is 0.0001 mol / L. The medicament for dissolving the tophus, provided by the invention, has a good dissolving effect and can be used for effectively dissolving the tophus within a short time. Acute gouty arthritis model rats verify that the joint swelling degree can be remarkably relieved by injecting the medicament provided by the invention. When the medicament is used for dissolving the tophus, pain and physical injury caused by physical operation to a patient can be relieved, the operation time and difficulty can be greatly reduced, and the medicament is good in safety, has feasibility and clinical application potential and is wide in application prospect.

Description

Technical Field

[0001] The invention relates to the technical field of tophi dissolving technology, in particular to a medicament for dissolving tophi and application thereof. Background Art

[0002] Gout is a purine metabolic disease caused by excessive intake of purine substances and impaired uric acid metabolism, and is directly related to hyperuricemia. With the change of lifestyle and dietary structure, the incidence of gout has increased year by year, especially in developed countries and urban areas. According to surveys, with the improvement of people's living standards, the intake of seafood and meat in daily diet has gradually increased, and the incidence of hyperuricemia and concurrent gouty arthritis in the Chinese population has continued to rise. Statistics show that the incidence of hyperuricemia and gout in my country is showing a trend of younger age.

[0003] For patients with chronic gout, tophi crystals are easily formed in the joint cavity. Tophi are mainly caused by inflammation caused by the deposition of uric acid and sodium urate crystals in the joints and surrounding tissues due to the increase of uric acid levels in the body. Its main component is monosodium urate, which has the characteristics of low solubility, slow dissolution rate and stable chemical properties.

[0004] At present, the main means of surgical treatment is physical resection, which causes pain, bleeding and tissue damage to patients, and cannot completely remove tophi. Especially for patients with severe gout with multiple scattered points, the surgical treatment process is quite painful, and the operation time is long and difficult; the residual scattered points of tophi may also become "seeds" for the continued growth of tophi, causing the re-formation of large tophi. Therefore, it is urgent to research and develop a new method or material to facilitate the removal of tophi and reduce damage at the same time. Summary of the invention

[0005] The purpose of the present invention is to provide a medicament for dissolving tophi and its application, so as to solve the problems existing in the above-mentioned prior art. The medicament for dissolving tophi provided by the present invention has good dissolving effect and can effectively dissolve tophi in a short time.

[0006] To achieve the above object, the present invention provides the following solutions:

[0007] The invention provides application of potassium carbonate in preparing a product for dissolving tophi.

[0008] Optionally, the concentration of potassium carbonate is 0.0001 mol / L.

[0009] The present invention also provides a medicament for dissolving tophi, wherein the medicament comprises potassium carbonate.

[0010] Optionally, the concentration of potassium carbonate is 0.0001 mol / L.

[0011] Optionally, the medicament further comprises auxiliary ingredients.

[0012] Optionally, the auxiliary component includes polyethylene glycol or DMSO.

[0013] Optionally, the dosage form of the medicament includes a liquid dosage form.

[0014] Examples of common specific situations in clinical applications and corresponding operable methods of the medicaments provided by the present invention are as follows:

[0015] For tophi outside the joints, the drug is injected into the lesion site. After a full response, the dissolution of the tophi can be monitored or observed through endoscopy or limited open surgery, depending on the situation. After confirming that the tophi are dissolved and removed, flushing and drainage are performed.

[0016] For intra-articular tophi, after the drug is injected into the joint cavity and fully reacted, the dissolution of the tophi can be monitored and observed through arthroscopy. If necessary, assisted endoscopic surgery can be performed to remove the residual tophi. After confirming that the tophi are dissolved and removed, flushing and drainage are performed.

[0017] For severe tophi with extensive erosion, partial tophi removal combined with multiple injections of drugs or immersion of the lesion site in drugs can be considered under the microscope or through limited open surgery. After the tophi are completely dissolved, flushing and drainage can be performed.

[0018] The present invention discloses the following technical effects:

[0019] The medicament for dissolving tophi provided by the present invention has a good dissolving effect and can effectively dissolve tophi in a short time. It has been verified by rats with acute gouty arthritis model that the degree of joint swelling can be significantly relieved by injection of the medicament provided by the present invention. In specific operations, the medicament can be injected into the lesion site, and after sufficient reaction, it can be discharged from the body through negative pressure drainage. It can not only dissolve local tophi, reduce the pain and physical damage caused to patients by physical operations, but also greatly reduce the operation time and difficulty, have good safety, feasibility and clinical application potential, and have broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0021] Figure 1 The dissolution results of sodium bicarbonate solutions with different concentrations;

[0022] Figure 2is the pH of sodium bicarbonate solution of different concentrations;

[0023] Figure 3 The dissolution results of sodium hydroxide solutions with different concentrations;

[0024] Figure 4 is the pH of sodium hydroxide solution with different concentrations;

[0025] Figure 5 The dissolution results of potassium dihydrogen phosphate solutions with different concentrations;

[0026] Figure 6 is the pH of potassium hydrogen phosphate solution with different concentrations;

[0027] Figure 7 The dissolution results of potassium dihydrogen phosphate solutions with different concentrations;

[0028] Figure 8 is the pH of potassium dihydrogen phosphate solution with different concentrations;

[0029] Fig. 9 The dissolution results of potassium chloride solutions with different concentrations;

[0030] Fig.10 is the pH of potassium chloride solution with different concentrations;

[0031] Fig.11 The dissolution results of potassium acetate solution with different concentrations;

[0032] Fig.12 is the pH of potassium acetate solution with different concentrations;

[0033] Fig.13 The dissolution results of potassium bicarbonate solutions with different concentrations;

[0034] Fig.14 is the pH of potassium bicarbonate solution of different concentrations;

[0035] Fig.15 The dissolution results of potassium carbonate solutions with different concentrations;

[0036] Fig.16 Results of the CCK8 cytotoxicity test;

[0037] Fig.17 This is a comparison picture of the legs of model rats;

[0038] Fig.18 This is a statistical chart of the changes in the ankle joint diameter of rats after drug administration. DETAILED DESCRIPTION

[0039] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0040] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0041] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0042] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0043] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0044] Example 1 Qualitative results of in vitro dissolution analysis of multiple groups of drugs

[0045] In the in vitro drug screening method, the screening process of drugs that can dissolve sodium urate in vitro includes qualitative tests and quantitative experiments. The main purpose is to explore drugs that can dissolve sodium urate in vitro and cause less damage to human tissues. The specific experimental methods are as follows:

[0046] Four groups of drug reaction systems were set up: 37°C, normal saline; 37°C, distilled water; 60°C, normal saline; 60°C, distilled water. For crystalline drugs (sodium bicarbonate, sodium carbonate, sodium monohydrogen phosphate, sodium dihydrogen phosphate, citric acid, sodium hydroxide), a 0.5M concentration of drug solution was prepared in a 100mL beaker, transferred to a magnetic stirrer, and stirred for 5 minutes to ensure that the drug was completely dissolved. Then, 0.1g of sodium urate was added to each reaction system as the target solute. In the three groups of liquid reagents (PBS buffer, limonene, hydrochloric acid), 500mL of PBS buffer, 200mL of limonene, and 40mL of hydrochloric acid diluted with a volume ratio of 1:12 were used. All drugs were placed in a water bath in advance for heating to ensure that the reaction system reached the expected temperature.

[0047] After consulting the data, the candidate drugs were dissolved in water into a uniform system, and then sodium urate was added to the clear and transparent solution. After continuous stirring and dissolution, there was no obvious dissolution of the solid in all systems. Under the action of the magnetic stirrer, the sodium urate solid was a stable white suspension. After being taken out of the water bath and left to stand for a long time, the sodium urate solid had no precipitation. After centrifugation in a centrifuge, the solid precipitated at the bottom of the centrifuge tube. The qualitative test results of various substances are shown in Table 1.

[0048] Table 1 Qualitative test results of various substances

[0049]

[0050] Control experiment: Analysis of the effect of the control group (distilled water / normal saline)

[0051] Before conducting a specific substance quantitative experiment, set up distilled water and physiological saline as control groups, weigh 100 mL of liquid into a beaker respectively, put it in a 37°C water bath for preheating, add 0.1 g of sodium urate, stir it at a constant temperature for a long time, and weigh the remaining substance after a period of reaction to obtain the amount of sodium urate dissolved in the control group.

[0052] In order to eliminate the effect of solvent on the solubility of sodium urate, the control group experiment used distilled water and saline dissolution systems, respectively, and the reaction results are shown in Table 2. After long-term stirring in distilled water, sodium urate solids were completely dissolved, and there was no white precipitate at the bottom of the centrifuge tube after centrifugation. This is consistent with the solubility of sodium urate in distilled water, but due to the weak dissolution kinetics of sodium urate, the dissolution rate is extremely slow, and it takes 35 minutes for 100 mg to be completely dissolved. This can also be compared with the above-mentioned drug dissolution experiment, and the upper limit of the dissolution reaction time is set based on this. In the saline reaction system, the sodium urate solid finally appeared as a white suspension, and after centrifugation, there were more white precipitates at the bottom. Compared with the distilled water group, the presence of sodium ions in the saline solution inhibited the dissolution of sodium urate solids. This also shows that the common ion effect has a more significant effect on the dissolution of sodium urate.

[0053] Therefore, in the sodium urate dissolution experiment, distilled water should be used as the solvent of the reaction system to avoid the influence of the solvent itself on the dissolution of sodium urate.

[0054] Table 2 Experimental results of control group

[0055]

[0056] Example 2 Quantitative results of in vitro dissolution analysis of multiple groups of drugs

[0057] Based on the chemical calculation of the existence form of sodium urate in solution and the solubility curve analysis under different pH values, alkaline solvents are preferred in this experiment. + In order to promote the dissolution of sodium urate, this experiment selected a variety of potassium salts and compared the dissolution differences among the groups.

[0058] According to the results of the control experiment in Example 1, a suitable reaction solvent is selected. In the experiment of each alternative drug, four different concentration gradients are set for each drug, and the corresponding solid mass under each gradient is weighed and added to a 150mL beaker to 100mL. The beaker is placed in a 37°C water bath and stirred to dissolve. After the solute is completely dissolved, the pH value of the solution is measured, 0.1g sodium urate is added to the preheated reaction beaker, and the reaction time is recorded. With the reaction time of the control experiment as the upper limit, after the reaction is completed, the beaker residue is divided into three centrifuge tubes of known mass. At 37°C, a speed of 10000r / min is used for centrifugation for 60min until the solid in the solution is completely precipitated and the upper solution is clear. Discard the supernatant, open the bottle mouth, put it in a drying oven at 60°C and dry it until the water is completely volatilized, weigh the final weight, and use the final remaining solid mass of the reaction system to compare the solid mass of each group to obtain the most suitable drug type and concentration.

[0059] 1. Na + Alkali and Na+ Effect of Salt on the Dissolution of Sodium Urate

[0060] Sodium bicarbonate reaction system: Since sodium bicarbonate has a low solubility, the concentrations of the four sodium bicarbonate reaction systems are: 0.25M, 0.5M, 0.75M, 1M. The pH values ​​of the solutions at each concentration are shown in Table 1. Figure 2 Compared with sodium hydroxide solution, the pH of the sodium bicarbonate reaction system is lower, presenting a neutral and weakly alkaline environment. Within the upper limit of the reference time of the control experiment, the sodium urate solid still presents an obvious white suspension, so the dissolution activity of sodium urate in it is poor and the dissolution amount is small ( Figure 1 ).

[0061] Sodium hydroxide reaction system: Based on the solubility of sodium hydroxide, four different concentrations were prepared with distilled water: 0.5M, 1M, 1.5M, and 2M. The pH values ​​of the solutions at each concentration are shown in Table 1. Figure 4 Sodium urate was completely dissolved in all four reaction systems ( Figure 3 ), and as the concentration of sodium hydroxide increases, the pH continues to rise, the dissolution time gradually decreases, and the dissolution rate gradually accelerates. This also confirms that sodium urate solid is more soluble in alkaline systems.

[0062] According to experiments, sodium urate has good solubility in strong alkaline solutions, and the sodium ion content in the solution should be reduced. Considering the internal environment, strong alkaline solutions cannot be directly used in the joint cavity, which can easily cause secondary damage to the injured joint.

[0063] 2. K + Base and K + Effect of Salt on the Dissolution of Sodium Urate

[0064] In the two systems of dipotassium hydrogen phosphate and potassium dihydrogen phosphate, four concentration systems of dipotassium hydrogen phosphate were set according to the solubility of the substance: 0.4M, 0.8M, 1.2M, 1.6M (the pH values ​​of the solutions of each concentration are shown in Figure 6 ) and three concentrations of potassium dihydrogen phosphate: 0.025M, 0.05M, 0.075M (see pH values ​​of solutions of each concentration Figure 8 After measurement, the pH of the solution system of potassium dihydrogen phosphate and potassium dihydrogen phosphate was acidic. After sufficient reaction, a large amount of solid remained in the beaker. The results were obtained by centrifugation and drying and weighing. The two substances had very little effect on the dissolution of sodium urate ( Figure 5 , Figure 7 ). This also confirms that in an acidic environment, it is not conducive for sodium urate to dissolve.

[0065] Four concentrations of potassium chloride: 0.5M, 1M, 1.5M, 2M. The solution is generally neutral ( Fig.10), compared with the results of potassium dihydrogen phosphate and dipotassium hydrogen phosphate groups, the dissolution quality of sodium urate increased slightly ( Fig. 9 ).

[0066] Potassium acetate reaction system: 0.5M, 1M, 1.25M, 1.5M, the solution is weakly alkaline ( Fig.12 ), compared with the above three groups of phosphates, the solubility of sodium urate in the system is greatly improved ( Fig.11 ). The experimental results of four groups, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, potassium chloride and potassium acetate, were compared and analyzed. The results preliminarily showed that K + In the distilled water system, the amount of sodium urate dissolved has a certain correlation with the pH of the solution. The higher the pH of the solution, the less the residual amount of sodium urate, which is conducive to the dissolution of sodium urate. + Compared with the experimental results of the group, in the same pH environment, K + The solution system is more conducive to the dissolution of sodium urate.

[0067] Potassium bicarbonate reaction system: 0.5M, 1M, 1.25M, 1.5M, the solution is weakly alkaline ( Fig.14 ), but the final result shows less dissolved sodium urate ( Fig.13 ).

[0068] Potassium carbonate reaction system: Potassium carbonate concentration and dissolution effect Fig.15 As shown in the figure, the dissolution effect is significant. As the concentration of potassium carbonate increases, the time required for the complete dissolution of sodium urate gradually decreases (among which, the time required for 0.0001 mol / L potassium carbonate solution to completely dissolve sodium urate is: 11 minutes). In the subsequent experiments in which the concentration of potassium carbonate solution was gradually reduced, there was no significant difference in the dissolution experiment of sodium urate as the pH gradually decreased, and there was no significant difference in the promotion of the dissolution of sodium urate by potassium carbonate solution as the pH of the solution gradually decreased.

[0069] 3. Mixture of potassium carbonate and polyethylene glycol

[0070] 100mL of 10 -3 mol / L or 10 -4 Add 5 mL of polyethylene glycol 200 solution to the 1.5 mol / L potassium carbonate solution, add 100 mg of sodium urate powder, and stir for 15 minutes in a 37°C water bath until the sodium urate powder is completely dissolved. Polyethylene glycol has good biocompatibility as a variety of drug preparations, controls drug diffusion, and improves drug efficacy, and is suitable as an auxiliary component of this dissolving agent.

[0071] 4. Mixture of potassium carbonate and DMSO

[0072] 100mL of 10 -3 mol / L or 10 -43 mL of DMSO was added to 100 mL of 1 mol / L potassium carbonate solution, and 100 mg of sodium urate powder was added. The mixture was stirred for 15 min in a 37°C water bath, and the sodium urate powder was completely dissolved. DMSO, as a permeability protective agent, can change the permeability of biofilm to drug preparations, and facilitate the potassium carbonate solution to fully dissolve irregular sodium urate powder in complex tissues, and is suitable as an auxiliary component of this dissolving agent.

[0073] Example 3 Cytotoxicity Test

[0074] 1. Cytotoxicity assay

[0075] After cryopreserved cells (mouse fibroblasts, purchased from Synbio (Shanghai) Biotechnology Co., Ltd.) were revived and passaged, the cell suspension was inoculated in a 96-well plate (200 μL / well, 2000 cells / well) and placed in a 37°C constant temperature incubator for culture. -4 mol / L potassium carbonate solution. After 1 day of cell culture, the culture medium was aspirated and complete culture medium was added as the control group. 10 -4 mol / L、10 -3 mol / L、10 -2 The cells were cultured in a constant temperature incubator for 1 d, 2 d, and 3 d, respectively. 110 μL CCK-8 reagent was added for incubation. The absorbance at 450 nm was measured using an ELISA reader. Graphpad prism software was used for data analysis to compare the significant differences among the groups.

[0076] 2. Cytotoxicity test results and analysis

[0077] The results are as follows Fig.16 As shown, the concentration used is 10 -2 mol / L potassium carbonate solution, culture cells for 24h, 48h, 72h, and measure absorbance at 460nm. Compared with the control group using complete culture medium, the cells showed stronger cytotoxicity and higher cell lethality. -3 mol / L potassium carbonate solution, the cells were severely damaged at 24h, and there was no significant difference between the results at 48h and 72h and the control group. -4 After culturing cells with 10 mol / L potassium carbonate solution, the cells were slightly damaged at 24 h, and the experimental results at 48 h and 72 h were not significantly different from those of the control group. -4 The toxicity of 1 mol / L potassium carbonate solution to cells was within an acceptable range and was suitable for subsequent in vivo experiments.

[0078] Example 4 Animal Modeling Verification

[0079] 1. Animal modeling experimental methods

[0080] Preparation of sodium urate crystal suspension: Take 0.1 g of sodium urate in a beaker, add 100 mL of distilled water, stir at 37°C until a milky white suspension is formed, and set aside for later use.

[0081] Preparation of anesthetics: Use chloral hydrate to prepare a chloral hydrate aqueous solution at a mass ratio of 1:10. Then transfer the solution to a 50 mL sterile centrifuge tube, wrap it with tin foil to protect it from light, and store it in a refrigerated environment at 4°C.

[0082] All rats were prepared with acute gouty arthritis rat model according to Coderre method: the rats were weighed and anesthetized with 1 mL of chloral hydrate injected intraperitoneally. When the rats were lying on their side and not moving, a 1 mL syringe was inserted into the ankle joint cavity from the outside and back of the ankle joint of the leg, and 0.2 mL of sodium urate suspension was injected. The rats were subjected to flexion and extension exercises for 4 min. At 24 h and 48 h, the modeling results were evaluated using the rat ankle inflammation index score (grading criteria see Table 3) to determine the modeling effect.

[0083] Table 3 Grading standards of inflammation index

[0084]

[0085] 2. In situ drug injection therapy and effect determination method

[0086] Preparation of potassium carbonate solution: Weigh 0.01 mol of potassium carbonate powder and dissolve it in water. Use distilled water to make up to 100 mL to obtain a potassium carbonate solution with a concentration of 1 mol / L. At the same time, prepare 3 beakers and use a measuring cylinder to measure 99 mL of distilled water. Take 1 mL from the 1 mol / L potassium carbonate solution and add it to 99 mL of water. Use a magnetic stirrer to stir and mix. Repeat the above steps to finally obtain a potassium carbonate solution with a concentration of 10. -4 mol / L potassium carbonate solution was transferred to a 50 mL sterile centrifuge tube.

[0087] After the gout in the ankle joint of the rats stabilized, the drug treatment was carried out. 0.2 mL of the prepared potassium carbonate solution was injected around the gout mass. After the animals were returned to the mouse room, the ankle diameter was measured with a vernier caliper at 0h, 24h, 48h, and 72h. The degree of joint swelling of the rats was calculated based on the change in the ankle diameter before and after the modeling. The joint diameter before and after drug administration was compared ( Fig.17 ) to preliminarily judge the therapeutic effect. The rats in this experiment used self-control, and the modeling effect and treatment effect were determined based on the changes in their joint diameters before and after, as well as the comparison of the left and right joints of the same rat.

[0088] 3. Changes in joint swelling (diameter) and effect analysis

[0089] The concentration is 10 -4 After the injection of mol / L potassium carbonate solution into the swollen joint of rats, the overall joint diameter tended to decrease (Table 4, Fig.18 ). The experimental results preliminarily verified that potassium carbonate solution has a therapeutic effect in rats with acute gouty arthritis. After 48 hours of administration, the ankle diameter was significantly reduced.

[0090] Table 4 Changes in ankle diameter of rats after administration

[0091]

[0092] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Application of potassium carbonate in the preparation of products for dissolving tophi.

2. The use according to claim 1, characterized in that The concentration of potassium carbonate is 0.0001 mol / L.

3. A medicament for dissolving tophi, characterized in that: The agent comprises potassium carbonate.

4. The medicament according to claim 3, characterized in that The concentration of potassium carbonate is 0.0001 mol / L.

5. The medicament according to claim 3, characterized in that The medicament may also contain auxiliary ingredients.

6. The agent according to claim 5, characterized in that The auxiliary components include polyethylene glycol or DMSO.

7. The medicament according to any one of claims 3 to 6, characterized in that The dosage form of the medicament includes a liquid dosage form.

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