A pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy and a preparation method thereof

Abstract

The present invention relates to the field of bioengineering pharmaceutical technology, and in particular to a pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy and a preparation method thereof, wherein the pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy is composed of a co-crystal formed by febuxostat, p-aminobenzoic acid and salicylic acid, and an emulsion system constructed by isopropyl stearate, polyethylene glycolized lecithin and modified hydroxyethyl cellulose. The present invention utilizes febuxostat, p-aminobenzoic acid and salicylic acid to form a co-crystal, optimizes drug solubility and bioavailability through a hydrogen bond network, enhances stability and reduces irritation. Polyethylene glycolized lecithin and carboxymethylated hydroxyethyl cellulose act synergistically, the former reduces interfacial tension and stabilizes the emulsion, and the latter enhances dispersion stability through hydrogen bonds, regulates the release rate, and achieves efficient sustained release.

Description

Technical Field

[0001] The present invention relates to the technical field of bioengineering pharmaceuticals, and in particular to a pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy and a preparation method thereof. Background Art

[0002] Hyperuricemia nephropathy, also known as gouty nephropathy or uric acid nephropathy, refers to a series of kidney diseases caused by the deposition of urate crystals in the kidneys due to high uric acid levels in the blood. This condition may be caused by excessive uric acid production or a decrease in the kidney's ability to excrete uric acid. Long-term uncontrolled hyperuricemia will gradually damage the kidneys and may lead to tubulointerstitial nephritis, uric acid stone formation, and even progress to chronic renal failure. Uric acid excretion promoters are drugs that can increase the excretion of uric acid by the kidneys and reduce the concentration of uric acid in the blood. Representative drugs include benzbromarone, probenecid, etc.; they inhibit the reabsorption of uric acid by the renal tubules, prompting more uric acid to be excreted from the body through urine. Uric acid synthesis inhibitors: These drugs reduce the production of uric acid by inhibiting the activity of xanthine oxidase. Allopurinol is the most common drug of this type. It can significantly reduce uric acid levels and is suitable for various types of hyperuricemia. Febuxostat, as a new generation of xanthine oxidase inhibitors, has higher selectivity and better safety. Drugs such as sodium bicarbonate can alkalize urine, increase the solubility of uric acid, prevent the formation of urate crystals, and thus reduce the burden on the kidneys. Some antihypertensive drugs, such as losartan and irbesartan, can not only control blood pressure, but also reduce uric acid levels through various pathways, which is beneficial to patients with hyperuricemia nephropathy. Although there are currently a variety of drugs available for the treatment of hyperuricemia nephropathy, there are still some technical challenges: many existing drug preparations have poor solubility, resulting in low bioavailability and poor absorption of drugs in the body, which affects the therapeutic effect. For example, allopurinol has poor solubility, which limits its absorption in the body. Based on the above problems, it is extremely necessary to develop a drug preparation that has excellent biocompatibility, can work efficiently and has low side effects to alleviate the progression of hyperuricemia nephropathy. Summary of the invention

[0003] In view of the defects of the prior art, the object of the present invention is to provide a pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy and a preparation method thereof.

[0004] The technical effect described in the present invention is achieved through the following technical scheme: a pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy, which is composed of a co-crystal formed by febuxostat, para-aminobenzoic acid and salicylic acid, and an emulsion system constructed by isopropyl stearate, polyethylene glycol lecithin and modified hydroxyethyl cellulose.

[0005] Preferably, the specific preparation steps of the polyethylene glycol lecithin are as follows:

[0006] A1: dissolving lecithin in 10 times the mass volume of dichloromethane, stirring and dissolving uniformly to obtain a lecithin solution; dissolving polyethylene glycol in 10 times the mass volume of dichloromethane, stirring and dissolving uniformly to obtain a polyethylene glycol solution;

[0007] A2: Add an activator prepared by using N,N'-dicyclohexylcarbodiimide and N-hydroxysuccinimide in a molar ratio of 1.1:1 to the polyethylene glycol solution prepared in step A1, and stir the mixture for 2 hours at room temperature to obtain an activated solution; then slowly add the activated solution to the lecithin solution prepared in step A1, and simultaneously add triethylamine, stir the mixture for 24 hours at room temperature, centrifuge, filter, treat with a rotary evaporator, wash with ether, and freeze-dry at -40°C in a vacuum until constant weight is obtained to obtain polyethylene glycolized lecithin.

[0008] Preferably, the mass ratio of lecithin and polyethylene glycol in step A2 is 2-3:1; the amount of the activator is 5% of the total mass of lecithin and polyethylene glycol; and the amount of triethylamine is 10% of the total mass of lecithin and polyethylene glycol.

[0009] Preferably, the specific preparation steps of the modified hydroxyethyl cellulose are as follows:

[0010] B1: Add hydroxyethyl cellulose to 10 times the mass volume of 30wt% NaOH solution three times, stir and dissolve evenly, then slowly add 10wt% sodium monochloroacetate solution, heat in a water bath to control the temperature at 50-80℃, react for 1-4h, and obtain a mixed solution;

[0011] B2: Add HCL solution to the mixed solution prepared in step B1 to adjust the pH to 6-7, stir evenly, add 0.5 times the volume of ethanol, let stand for 6-12 hours, centrifuge, filter, wash three times with ethanol, and vacuum dry at 50-60°C to constant weight to obtain modified hydroxyethyl cellulose.

[0012] Preferably, the ratio of the sodium monochloroacetate solution to hydroxyethyl cellulose in step B1 is 10 mL: 1 g.

[0013] Preferably, the preparation method of the pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy comprises the following specific preparation steps:

[0014] S1: Dissolve febuxostat, p-aminobenzoic acid and salicylic acid in anhydrous ethanol respectively, heat in a water bath and stir to dissolve evenly, obtain febuxostat solution, p-aminobenzoic acid solution and salicylic acid solution, slowly pour the dissolved febuxostat solution into the p-aminobenzoic acid solution, stir to mix evenly, and obtain a mixed solution; pour the mixed solution into the dissolved salicylic acid solution, add 0.1% polyvinyl pyrrolidone, heat in a water bath and gradually increase the temperature to 65-75°C at 5°C / min, then maintain the temperature and stir for 60-100 minutes, naturally cool to room temperature, stand for 6-12 hours, filter, and wash with cold deionized water to obtain a eutectic;

[0015] S2: heating isopropyl stearate to 50-60° C. and completely melting it to obtain pretreated isopropyl stearate; adding polyethylene glycol lecithin to deionized water, heating the temperature in a water bath to 50-60° C., stirring to dissolve evenly, and then slowly adding the pretreated isopropyl stearate and vitamin E, homogenizing to form a uniform emulsion, and continuously stirring until it is naturally cooled to room temperature to obtain a cooled emulsion;

[0016] S3: adding the eutectic prepared in step S1 to anhydrous ethanol, stirring and dissolving uniformly to obtain a eutectic solution; then slowly adding the eutectic solution to the cooled emulsion prepared in step S2, stirring until the eutectic is uniformly distributed in the emulsion, and then slowly adding modified hydroxyethyl cellulose within 10 minutes, stirring until the modified hydroxyethyl cellulose is completely dissolved, and adjusting the pH to 6-6.5 with sodium citrate and citric acid to obtain a dispersed emulsion;

[0017] S4: After the dispersed emulsion in step S3 is stirred at 500-1000 rpm for 10-30 min using a magnetic stirrer, the homogenization cycle is performed 3-5 times, the parameters are set to 300-500 bar, the temperature is room temperature, and each treatment is 2 min. After the homogenization treatment is completed, it is stirred again at 500-1000 rpm for 10 min to obtain an emulsified pharmaceutical preparation.

[0018] Preferably, the ratio of the amount of febuxostat, p-aminobenzoic acid, salicylic acid and anhydrous ethanol used in step S1 is 1 mg:1.5 mg:1 mg:10 mL.

[0019] Preferably, the ratio of the amount of isopropyl stearate, polyethylene glycol lecithin, vitamin E and deionized water in step S2 is 5-10g:1g:50mL:0.1-0.3g; the specific parameters of the homogenization treatment are an initial homogenization pressure of 500-800bar, a temperature of 40-60°C, a homogenization treatment time of 2-3min each time, and a total of 3 cycles. During the second homogenization treatment, the homogenization pressure is increased to 1000-1200bar, and the other parameters are the same as the first treatment. During the third homogenization treatment, the temperature is lowered to 20°C, and the other parameters are the same as the first treatment.

[0020] Preferably, the ratio of the amount of the eutectic, modified hydroxyethyl cellulose, cooling emulsion and anhydrous ethanol used in step S3 is 50 mg: 1-2 g: 100 mL: 50-100 mL.

[0021] The beneficial effects of the present invention are as follows:

[0022] The present invention utilizes the formation of hydrogen bonds between the carboxyl group (-COOH) of Febuxostat and the amino group (-NH2) of para-aminobenzoic acid and the hydroxyl group (-OH) of salicylic acid. The formation of co-crystals can improve the solubility of poorly soluble drugs. This is because the hydrogen bond network formed between the drug molecules and the co-former can change the arrangement and exposure of the drug molecules, thereby affecting their dissolution behavior; the improvement of solubility is directly related to the absorption rate of the drug, that is, the bioavailability, and the co-crystal can make the absorption of Febuxostat in the body more rapid and complete. In addition, the structural stability of the co-crystal is enhanced by hydrogen bonds and other intermolecular forces, which effectively prevents the degradation of the drug under conditions of high temperature and high humidity, and protects the drug from the influence of humidity, light and oxidation; the co-crystal reduces the close packing of molecules in the crystal through the formation of a new lattice structure, which can significantly improve the solubility of the drug in water, and helps the subsequent oil phase and water to combine to construct an emulsion. The co-crystal structure can reduce irritation to organisms. Febuxostat, para-aminobenzoic acid and salicylic acid are all known biocompatible compounds. The degradation products of the co-crystals they form in the human body are also safe and will not cause allergies or other adverse reactions. Co-crystals change the crystal form of drugs, while emulsions disperse drugs in nano-scale droplets. Both can promote the absorption of drugs in the body, improve bioavailability, and thus enhance drug efficacy.

[0023] The present invention utilizes polyethylene glycol lecithin and carboxymethylated hydroxyethyl cellulose to synergistically act on the emulsion preparation of febuxostat, p-aminobenzoic acid and salicylic acid co-crystal. The polyethylene glycol lecithin forms a stable film at the oil-water interface through its amphiphilic structure, reduces the oil-water interfacial tension, forms fine and uniform oil droplets, and then improves the stability of the emulsion. The polyethylene glycol coating can reduce the recognition of macrophages, reduce the clearance rate in the body, effectively prolong the residence time of the emulsion in the blood circulation, and improve the bioavailability of the drug. The carboxymethylated hydroxyethyl cellulose interacts with the active groups in the febuxostat, p-aminobenzoic acid and salicylic acid co-crystal through hydrogen bonding, further stabilizes the dispersion state of the co-crystal in the emulsion, increases the viscosity of the solution, prevents the aggregation and phase separation of oil droplets, and enhances the overall stability of the emulsion. The polyethylene glycol lecithin provides good biocompatibility, forms a preliminary stable emulsion, reduces the clearance rate of emulsion particles in the body, and improves the effectiveness of the drug. Carboxymethylated hydroxyethyl cellulose forms a viscous protective layer to protect the drug from rapid metabolism and degradation, while adjusting the release rate of the drug by increasing viscosity to achieve an effective sustained-release effect. PEGylated lecithin ensures uniform distribution of oil droplets, giving the emulsion a good appearance and texture, while carboxymethylated hydroxyethyl cellulose increases the fluidity and touch of the emulsion by increasing viscosity. The combination of the two significantly improves the physical properties of the emulsion. PEGylated lecithin ensures uniform dispersion of drug co-crystals in the emulsion, and carboxymethylated hydroxyethyl cellulose further prevents drug sedimentation by increasing viscosity and forming a protective layer, ensuring the stable presence of the drug in the emulsion. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings in the following description are only for the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0025] Figure 1 It is a biocompatibility test diagram of the emulsion preparations prepared in Examples 1 to 4 of the present invention and Comparative Examples 1 to 2;

[0026] Figure 2 is the FTIR infrared spectrum of the emulsion preparation prepared in Example 2 of the present invention;

[0027] Figure 3 is the XRD diffraction pattern of the co-crystal prepared in Example 2 of the present invention;

[0028] Figure 4 It is a dissolution test graph of the co-crystals prepared in Examples 2 to 3 of the present invention and Comparative Examples 1 to 2;

[0029] Figure 5It is a pH change diagram of the stability test of the emulsion preparations prepared in Examples 2 to 3 of the present invention and Comparative Examples 1 to 2. DETAILED DESCRIPTION

[0030] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. It should be noted that, unless otherwise specified, the raw materials involved in the present invention are purchased through conventional commercial channels.

[0031] Example 1: A pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy, comprising a co-crystal formed by febuxostat, p-aminobenzoic acid and salicylic acid, and an emulsion system constructed by isopropyl stearate, polyethylene glycol lecithin and modified hydroxyethyl cellulose.

[0032] 1. Preparation of PEGylated lecithin:

[0033] A1: Dissolve 20 g of lecithin in 200 mL of dichloromethane, stir and dissolve evenly to obtain a lecithin solution; dissolve 10 g of polyethylene glycol in 100 mL of dichloromethane, stir and dissolve evenly to obtain a polyethylene glycol solution;

[0034] A2: Add 1.5 g of an activator prepared by mixing N,N'-dicyclohexylcarbodiimide and N-hydroxysuccinimide in a molar ratio of 1.1:1 to the polyethylene glycol solution prepared in step A1, and stir the mixture for 2 h at room temperature to obtain an activated solution; then slowly add the activated solution to the lecithin solution prepared in step A1, and simultaneously add 3 g of triethylamine, stir the mixture for 24 h at room temperature, centrifuge, filter, treat with a rotary evaporator, wash with ether, and freeze-dry at -40°C in a vacuum until constant weight is obtained to obtain polyethylene glycolized lecithin;

[0035] 2. Preparation of modified hydroxyethyl cellulose:

[0036] B1: 10 g of hydroxyethyl cellulose was added to 100 mL of 30 wt% NaOH solution in three times, and after being stirred and dissolved evenly, 100 mL of 10 wt% sodium monochloroacetate solution was slowly added, and the temperature was controlled at 50°C in a water bath, and the reaction was carried out for 4 hours to obtain a mixed solution;

[0037] B2: Add HCL solution to the mixed solution prepared in step B1 to adjust the pH to 7, stir evenly, add 100 mL of ethanol, let stand for 6 hours, centrifuge, filter, wash three times with ethanol, and dry under vacuum at 50° C. to constant weight to obtain modified hydroxyethyl cellulose;

[0038] 3. Preparation of pharmaceutical preparations for alleviating the progression of hyperuricemia nephropathy:

[0039] S1: Dissolve 10 mg of febuxostat, 15 mg of p-aminobenzoic acid and 10 mg of salicylic acid in 100 mL of anhydrous ethanol respectively, heat in a water bath and stir to dissolve evenly to obtain a febuxostat solution, a p-aminobenzoic acid solution and a salicylic acid solution, slowly pour the dissolved febuxostat solution into the p-aminobenzoic acid solution, stir and mix evenly to obtain a mixed solution; pour the mixed solution into the dissolved salicylic acid solution, add 0.01 g of polyvinyl pyrrolidone, heat in a water bath and gradually increase the temperature to 65°C at 5°C / min, then maintain the temperature and stir for 100 minutes, naturally cool to room temperature, stand for 6 hours, filter, and wash with cold deionized water to obtain a eutectic;

[0040] S2: 10 g of isopropyl stearate was heated to 50°C and completely melted to obtain pretreated isopropyl stearate; 2 g of polyethylene glycol lecithin was added to 100 mL of deionized water, heated to 50°C in a water bath, stirred and dissolved evenly, and then the pretreated isopropyl stearate and 0.2 g of vitamin E were slowly added for homogenization. The specific parameters were an initial homogenization pressure of 500 bar, a temperature of 40°C, and a homogenization time of 2 min. A total of 3 cycles were performed. The homogenization pressure was increased to 1000 bar for the second homogenization, and the other parameters were the same as the first treatment. The temperature was lowered to 20°C for the third homogenization, and the other parameters were the same as the first treatment. After a uniform emulsion was formed, stirring was continued until it was naturally cooled to room temperature to obtain a cooled emulsion;

[0041] S3: adding 50 mg of the eutectic prepared in step S1 to 50 mL of anhydrous ethanol, stirring and dissolving uniformly to obtain a eutectic solution; then slowly adding the eutectic solution to 100 mL of the cooled emulsion prepared in step S2, stirring until the eutectic is uniformly distributed in the emulsion, and then slowly adding 1 g of modified hydroxyethyl cellulose within 10 min, stirring until the modified hydroxyethyl cellulose is completely dissolved, and adjusting the pH to 6 with sodium citrate and citric acid to obtain a dispersed emulsion;

[0042] S4: After the dispersed emulsion in step S3 was stirred at 500 rpm for 30 min using a magnetic stirrer, the homogenization cycle was performed 3 times, the parameters were set to 500 bar, the temperature was room temperature, and each treatment was 2 min. After the homogenization treatment was completed, it was stirred again at 500 rpm for 10 min to obtain an emulsified drug preparation.

[0043] Example 2: A pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy, comprising a co-crystal formed by febuxostat, p-aminobenzoic acid and salicylic acid, and an emulsion system constructed by isopropyl stearate, polyethylene glycol lecithin and modified hydroxyethyl cellulose.

[0044] 1. Preparation of PEGylated lecithin:

[0045] A1: Dissolve 28 g of lecithin in 280 mL of dichloromethane, stir and dissolve evenly to obtain a lecithin solution; dissolve 10 g of polyethylene glycol in 100 mL of dichloromethane, stir and dissolve evenly to obtain a polyethylene glycol solution;

[0046] A2: Add 1.9 g of an activator prepared by mixing N,N'-dicyclohexylcarbodiimide and N-hydroxysuccinimide in a molar ratio of 1.1:1 to the polyethylene glycol solution prepared in step A1, and stir the mixture for 2 h at room temperature to obtain an activated solution; then slowly add the activated solution to the lecithin solution prepared in step A1, and simultaneously add 3.8 g of triethylamine, stir the mixture for 24 h at room temperature, centrifuge, filter, treat with a rotary evaporator, wash with ether, and freeze-dry at -40°C in a vacuum until constant weight is obtained to obtain polyethylene glycolized lecithin;

[0047] 2. Preparation of modified hydroxyethyl cellulose:

[0048] B1: 10 g of hydroxyethyl cellulose was added to 100 mL of 30 wt% NaOH solution in three times, and after being stirred and dissolved evenly, 100 mL of 14 wt% sodium monochloroacetate solution was slowly added, and the temperature was controlled at 60°C in a water bath, and the reaction was carried out for 4 hours to obtain a mixed solution;

[0049] B2: Add HCL solution to the mixed solution prepared in step B1 to adjust the pH to 6.9, stir evenly, add 100 mL of ethanol, let stand for 10 hours, centrifuge, filter, wash three times with ethanol, and dry under vacuum at 60° C. to constant weight to obtain modified hydroxyethyl cellulose;

[0050] 3. Preparation of pharmaceutical preparations for alleviating the progression of hyperuricemia nephropathy:

[0051] S1: Dissolve 10 mg of febuxostat, 15 mg of p-aminobenzoic acid and 10 mg of salicylic acid in 100 mL of anhydrous ethanol respectively, heat in a water bath and stir to dissolve evenly to obtain a febuxostat solution, a p-aminobenzoic acid solution and a salicylic acid solution, slowly pour the dissolved febuxostat solution into the p-aminobenzoic acid solution, stir and mix evenly to obtain a mixed solution; pour the mixed solution into the dissolved salicylic acid solution, add 0.01 g of polyvinyl pyrrolidone, heat in a water bath and gradually increase the temperature to 70°C at 5°C / min, then maintain the temperature and stir for 90 minutes, naturally cool to room temperature, stand for 10 hours, filter, and wash with cold deionized water to obtain a eutectic;

[0052] S2: 18g of isopropyl stearate was heated to 55°C and completely melted to obtain pretreated isopropyl stearate; 2g of polyethylene glycol lecithin was added to 100mL of deionized water, heated to 55°C in a water bath, stirred and dissolved evenly, and then the pretreated isopropyl stearate and 0.5g of vitamin E were slowly added for homogenization. The specific parameters were an initial homogenization pressure of 700bar, a temperature of 50°C, and a homogenization time of 3min. A total of 3 cycles were performed. The homogenization pressure was increased to 1200bar for the second homogenization, and the other parameters were the same as the first treatment. The temperature was lowered to 20°C for the third homogenization, and the other parameters were the same as the first treatment. After a uniform emulsion was formed, stirring was continued until it was naturally cooled to room temperature to obtain a cooled emulsion;

[0053] S3: adding 50 mg of the eutectic prepared in step S1 to 85 mL of anhydrous ethanol, stirring and dissolving uniformly to obtain a eutectic solution; then slowly adding the eutectic solution to 100 mL of the cooled emulsion prepared in step S2, stirring until the eutectic is uniformly distributed in the emulsion, and then slowly adding 1.8 g of modified hydroxyethyl cellulose within 10 min, stirring until the modified hydroxyethyl cellulose is completely dissolved, and adjusting the pH to 6.5 with sodium citrate and citric acid to obtain a dispersed emulsion;

[0054] S4: After the dispersed emulsion in step S3 was stirred at 800 rpm for 20 min using a magnetic stirrer, the homogenization cycle was performed 5 times, the parameters were set to 400 bar, the temperature was room temperature, and each treatment was 2 min. After the homogenization treatment was completed, it was stirred again at 800 rpm for 10 min to obtain an emulsified drug preparation.

[0055] Example 3: A pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy, comprising a co-crystal formed by febuxostat, p-aminobenzoic acid and salicylic acid, and an emulsion system constructed by isopropyl stearate, polyethylene glycol lecithin and modified hydroxyethyl cellulose.

[0056] 1. Preparation of PEGylated lecithin:

[0057] A1: Dissolve 25 g of lecithin in 250 mL of dichloromethane, stir and dissolve evenly to obtain a lecithin solution; dissolve 10 g of polyethylene glycol in 100 mL of dichloromethane, stir and dissolve evenly to obtain a polyethylene glycol solution;

[0058] A2: Add 1.75 g of an activator prepared by mixing N,N'-dicyclohexylcarbodiimide and N-hydroxysuccinimide in a molar ratio of 1.1:1 to the polyethylene glycol solution prepared in step A1, and stir the mixture for 2 h at room temperature to obtain an activated solution; then slowly add the activated solution to the lecithin solution prepared in step A1, and simultaneously add 3.5 g of triethylamine, stir the mixture for 24 h at room temperature, centrifuge, filter, treat with a rotary evaporator, wash with ether, and freeze-dry at -40°C in a vacuum until constant weight is obtained to obtain polyethylene glycolized lecithin;

[0059] 2. Preparation of modified hydroxyethyl cellulose:

[0060] B1: 10 g of hydroxyethyl cellulose was added to 100 mL of 30 wt% NaOH solution in three times, and after being stirred and dissolved evenly, 100 mL of 12 wt% sodium monochloroacetate solution was slowly added, and the temperature was controlled at 60°C in a water bath, and the reaction was carried out for 3 hours to obtain a mixed solution;

[0061] B2: Add HCL solution to the mixed solution prepared in step B1 to adjust the pH to 6.5, stir evenly, add 100 mL of ethanol, let stand for 10 h, centrifuge, filter, wash three times with ethanol, and vacuum dry at 55° C. to constant weight to obtain modified hydroxyethyl cellulose;

[0062] 3. Preparation of pharmaceutical preparations for alleviating the progression of hyperuricemia nephropathy:

[0063] S1: Dissolve 10 mg of febuxostat, 15 mg of p-aminobenzoic acid and 10 mg of salicylic acid in 100 mL of anhydrous ethanol respectively, heat in a water bath and stir to dissolve evenly to obtain a febuxostat solution, a p-aminobenzoic acid solution and a salicylic acid solution, slowly pour the dissolved febuxostat solution into the p-aminobenzoic acid solution, stir and mix evenly to obtain a mixed solution; pour the mixed solution into the dissolved salicylic acid solution, add 0.01 g of polyvinyl pyrrolidone, heat in a water bath and gradually increase the temperature to 70°C at 5°C / min, then maintain the temperature and stir for 80 minutes, naturally cool to room temperature, let stand for 8 hours, filter, wash with cold deionized water to obtain a eutectic;

[0064] S2: 15 g of isopropyl stearate was heated to 52° C. and completely melted to obtain pretreated isopropyl stearate; 2 g of polyethylene glycolized lecithin was added to 100 mL of deionized water, heated to 52° C. in a water bath, stirred and dissolved evenly, and then the pretreated isopropyl stearate and 0.4 g of vitamin E were slowly added for homogenization. The specific parameters were an initial homogenization pressure of 600 bar, a temperature of 50° C., and a homogenization time of 3 min. A total of 3 cycles were performed. The homogenization pressure was increased to 1100 bar for the second homogenization, and the other parameters were the same as the first treatment. The temperature was lowered to 20° C. for the third homogenization, and the other parameters were the same as the first treatment. After a uniform emulsion was formed, stirring was continued until it was naturally cooled to room temperature to obtain a cooled emulsion;

[0065] S3: adding 50 mg of the eutectic prepared in step S1 to 75 mL of anhydrous ethanol, stirring to dissolve evenly, to obtain a eutectic solution; then slowly adding the eutectic solution to 100 mL of the cooled emulsion prepared in step S2, stirring until the eutectic is evenly distributed in the emulsion, and then slowly adding 1.5 g of modified hydroxyethyl cellulose within 10 min, stirring until the modified hydroxyethyl cellulose is completely dissolved, and adjusting the pH to 6.2 with sodium citrate and citric acid to obtain a dispersed emulsion;

[0066] S4: After the dispersed emulsion in step S3 was stirred at 600 rpm for 20 min using a magnetic stirrer, the homogenization cycle was performed 4 times, the parameters were set to 400 bar, the temperature was room temperature, and each treatment was 2 min. After the homogenization treatment was completed, it was stirred again at 700 rpm for 10 min to obtain an emulsified drug preparation.

[0067] Example 4: A pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy, comprising a co-crystal formed by febuxostat, p-aminobenzoic acid and salicylic acid, and an emulsion system constructed by isopropyl stearate, polyethylene glycol lecithin and modified hydroxyethyl cellulose.

[0068] 1. Preparation of PEGylated lecithin:

[0069] A1: Dissolve 30 g of lecithin in 300 mL of dichloromethane, stir and dissolve evenly to obtain a lecithin solution; dissolve 10 g of polyethylene glycol in 100 mL of dichloromethane, stir and dissolve evenly to obtain a polyethylene glycol solution;

[0070] A2: Add 2 g of an activator prepared by mixing N,N'-dicyclohexylcarbodiimide and N-hydroxysuccinimide in a molar ratio of 1.1:1 to the polyethylene glycol solution prepared in step A1, and stir the mixture for 2 h at room temperature to obtain an activated solution; then slowly add the activated solution to the lecithin solution prepared in step A1, and simultaneously add 4 g of triethylamine, stir the mixture for 24 h at room temperature, centrifuge, filter, treat with a rotary evaporator, wash with ether, and freeze-dry at -40°C in a vacuum until constant weight is obtained to obtain polyethylene glycolized lecithin;

[0071] 2. Preparation of modified hydroxyethyl cellulose:

[0072] B1: 10 g of hydroxyethyl cellulose was added to 100 mL of 30 wt% NaOH solution in three times, and after being stirred and dissolved evenly, 100 mL of 15 wt% sodium monochloroacetate solution was slowly added, and the temperature was controlled at 80°C in a water bath, and the reaction was carried out for 1 hour to obtain a mixed solution;

[0073] B2: Add HCL solution to the mixed solution prepared in step B1 to adjust the pH to 7, stir evenly, add 100 mL of ethanol, let stand for 12 hours, centrifuge, filter, wash with ethanol three times, and vacuum dry at 60° C. to constant weight to obtain modified hydroxyethyl cellulose;

[0074] 3. Preparation of pharmaceutical preparations for alleviating the progression of hyperuricemia nephropathy:

[0075] S1: Dissolve 10 mg of febuxostat, 15 mg of p-aminobenzoic acid and 10 mg of salicylic acid in 100 mL of anhydrous ethanol respectively, heat in a water bath and stir to dissolve evenly to obtain a febuxostat solution, a p-aminobenzoic acid solution and a salicylic acid solution, slowly pour the dissolved febuxostat solution into the p-aminobenzoic acid solution, stir to mix evenly to obtain a mixed solution; pour the mixed solution into the dissolved salicylic acid solution, add 0.01 g of polyvinyl pyrrolidone, heat in a water bath and gradually increase the temperature to 75°C at 5°C / min, then maintain the temperature and stir for 60 minutes, naturally cool to room temperature, stand for 12 hours, filter, and wash with cold deionized water to obtain a eutectic;

[0076] S2: 20 g of isopropyl stearate was heated to 60°C and completely melted to obtain pretreated isopropyl stearate; 2 g of polyethylene glycol lecithin was added to 100 mL of deionized water, heated to 60°C in a water bath, stirred and dissolved evenly, and then the pretreated isopropyl stearate and 0.6 g of vitamin E were slowly added for homogenization. The specific parameters were an initial homogenization pressure of 800 bar, a temperature of 60°C, and a homogenization time of 3 min. A total of 3 cycles were performed. The homogenization pressure was increased to 1200 bar for the second homogenization, and the other parameters were the same as the first treatment. The temperature was lowered to 20°C for the third homogenization, and the other parameters were the same as the first treatment. After a uniform emulsion was formed, stirring was continued until it was naturally cooled to room temperature to obtain a cooled emulsion;

[0077] S3: adding 50 mg of the eutectic prepared in step S1 to 100 mL of anhydrous ethanol, stirring and dissolving uniformly to obtain a eutectic solution; then slowly adding the eutectic solution to 100 mL of the cooled emulsion prepared in step S2, stirring until the eutectic is uniformly distributed in the emulsion, and then slowly adding 2 g of modified hydroxyethyl cellulose within 10 min, stirring until the modified hydroxyethyl cellulose is completely dissolved, and adjusting the pH to 6.5 with sodium citrate and citric acid to obtain a dispersed emulsion;

[0078] S4: After the dispersed emulsion in step S3 is stirred at 1000 rpm for 10 min using a magnetic stirrer, the homogenization cycle is performed 5 times, the parameters are set to 500 bar, the temperature is room temperature, and each treatment is 2 min. After the homogenization treatment is completed, it is stirred again at 1000 rpm for 10 min to obtain an emulsified drug preparation.

[0079] Comparative Example 1: The operation of Comparative Example 1 is basically the same as that of Example 2, except that febuxostat is used in Comparative Example 1 instead of the co-crystal formed by febuxostat, p-aminobenzoic acid and salicylic acid.

[0080] Comparative Example 2: The operation of Comparative Example 2 is basically the same as that of Example 3, except that lecithin is used in Comparative Example 2 instead of polyethylene glycolized lecithin.

[0081] Performance Testing:

[0082] Biocompatibility test: 10 μL of the emulsions prepared in Examples 1 to 4 and Comparative Examples 1 to 2 were placed on the bottom of the culture plate and evenly distributed in a 96-well plate with 8 wells per group of samples. 50 μL of DMEM medium was added to each well, and then HK-2 human renal epithelial cells were diluted to 1×10 5 / mL and then inoculate 50 μL in the wells added with samples. At the same time, 8 wells without samples were used as negative control groups, and 8 wells added with 10wt% dimethyl sulfoxide solution were used as positive control groups. The temperature was set at 37°C and 5% CO2. 5 μL of 5 mg / mL MTT solution was added after culturing for 24h, 72h, and 168h, respectively. After further culturing for 4h, the culture medium in the wells was aspirated, 50 μL of dimethyl sulfoxide was added to the wells, and the absorbance (OD value) was measured at 490nm using a microplate reader to measure the activity percentage [cell survival rate (%) = (OD value of the embodiment - OD value of the negative control group) / (OD value of the positive control group - OD value of the negative control group) × 100%], blank control group: only containing culture medium and MTT solution; negative control group: untreated cells were used as control; each sample was processed and analyzed three times, and the results were averaged. The results are shown in Figure 1 shown.

[0083] Depend on Figure 1 The results show that the emulsion preparation prepared by the present invention has good biocompatibility and does not cause toxic damage to cells. From the results of Comparative Example 1 and Example 2, it can be seen that the structure of the co-crystal effectively improves the biocompatibility of the preparation; from the results of Comparative Example 2 and Example 2, it can be seen that after lecithin replaces PEGylated lecithin, its biocompatibility is affected to a certain extent.

[0084] Spectrum test: The emulsion prepared in Example 2 was subjected to FTIR infrared spectrum test, and the results were as follows: Figure 2 The co-crystal sample prepared in Example 2 and the powder of fesitab were subjected to X-ray powder diffraction test, and the results were as follows: Figure 3 shown.

[0085] Depend on Figure 2 The results show that the emulsion preparation prepared by the present invention has obvious P=O stretching vibration peak of PEGylated lecithin and C=O stretching vibration peak of ester group of emulsification system, and the emulsifier solution is effectively prepared; and Figure 3 The results show that the co-crystal prepared by the present invention has an obvious new peak and effectively forms a new crystal structure.

[0086] Drug dissolution test: 3 samples of the cocrystal samples and febuxostat samples prepared in Examples 2 to 3 of the present invention were taken respectively, for a total of 18 samples, each sample weighing 100 mg, and each sample was dissolved in 500 mL of test solution (495 mL of pH 1.2 phosphate buffer + 5 mL of Tween-80), and samples were taken at 15 min, 30 min, 60 min, 90 min, 120 min, and 180 min, respectively. 10 mL of sample was taken at each sampling point, and then 10 mL of solution was added to measure the dissolution rate (X n’ =X n +(X1+X2+Xn-1 )V2 / V1; where X n is the relative percentage dissolution measured after correction for the nth time, X n’ is the relative percentage dissolution actually measured for the nth time, V1 is the volume of the dissolution medium, and V2 is the volume added after each sampling). The results are as follows Figure 4 shown.

[0087] Depend on Figure 4 The results show that the co-crystal prepared by the present invention can be significantly dissolved by more than 60% in 60 minutes, the dissolution effect is excellent, and the solubility is significantly higher than that of febuxostat.

[0088] Stability test: The emulsion samples prepared in Examples 2 to 3 of the present invention and Comparative Examples 1 to 2 were sealed in commercial packaging and placed at a temperature of 35 ± 5°C and a relative humidity of 75 ± 5% for 6 months. Samples were taken at the 1st, 2nd, 3rd and 6th month of the test period to test the solution pH. The average value of the three samples was taken. The results are shown in Table 1. Figure 5 shown.

[0089] Depend on Figure 5 The results show that the emulsion samples prepared by the present invention did not undergo significant changes in the 6-month long-term storage experiment, the pH fluctuated within a very small range, and the samples could be effectively preserved for a long time; from the results of Comparative Example 2 and Example 3, it can be seen that when lecithin replaces polyethylene glycolized lecithin, its stability is significantly reduced after long-term storage and high shear due to the lack of additional stabilization provided by the polyethylene glycol chain.

[0090] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy, characterized in that: The pharmaceutical preparation is composed of a co-crystal formed by febuxostat, p-aminobenzoic acid and salicylic acid, and an emulsion system constructed by isopropyl stearate, polyethylene glycol lecithin and modified hydroxyethyl cellulose, and the mass dosage ratio of febuxostat to p-aminobenzoic acid and salicylic acid is 1:1.5:1; The specific preparation steps of the polyethylene glycol lecithin are as follows: A1: dissolving lecithin in dichloromethane, stirring and dissolving uniformly to obtain a lecithin solution; dissolving polyethylene glycol in dichloromethane, stirring and dissolving uniformly to obtain a polyethylene glycol solution; A2: adding an activator prepared by mixing N,N'-dicyclohexylcarbodiimide and N-hydroxysuccinimide in a molar ratio of 1.1:1 to the polyethylene glycol solution prepared in step A1, stirring and reacting at room temperature to obtain an activated solution; then slowly adding the activated solution to the lecithin solution prepared in step A1, adding triethylamine at the same time, stirring and reacting at room temperature, centrifuging, filtering, treating with a rotary evaporator, washing with ether, and vacuum freeze-drying to constant weight to obtain polyethylene glycol lecithin; The mass ratio of lecithin and polyethylene glycol in step A2 is 2-3:1; the amount of the activator is 5% of the total mass of lecithin and polyethylene glycol; the amount of triethylamine is 10% of the total mass of lecithin and polyethylene glycol; The specific preparation steps of the modified hydroxyethyl cellulose are as follows: B1: Add hydroxyethyl cellulose to NaOH solution three times, stir and dissolve evenly, then slowly add sodium monochloroacetate solution, heat in a water bath to control the temperature, and after the reaction is completed, obtain a mixed solution; B2: Adding HCL solution to the mixed solution prepared in step B1 to adjust the pH, stirring evenly, adding ethanol, standing, centrifuging, filtering, washing, and vacuum drying to constant weight to obtain modified hydroxyethyl cellulose; The ratio of the sodium monochloroacetate solution to hydroxyethyl cellulose in step B1 is 10 mL: 1 g.

2. A method for preparing a pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy according to claim 1, characterized in that: The preparation steps of the pharmaceutical preparation are as follows: S1: Dissolve febuxostat, p-aminobenzoic acid and salicylic acid in anhydrous ethanol respectively, heat in a water bath and stir to dissolve evenly to obtain a febuxostat solution, a p-aminobenzoic acid solution and a salicylic acid solution, slowly pour the dissolved febuxostat solution into the p-aminobenzoic acid solution, stir and mix evenly to obtain a mixed solution; The mixed solution was poured into the dissolved salicylic acid solution, and then polyvinyl pyrrolidone was added, and the temperature was gradually increased to 65-75°C at 5°C / min in a water bath, and then the temperature was maintained and stirred for reaction. After the reaction was completed, the mixture was naturally cooled to room temperature, allowed to stand, filtered, and washed with cold deionized water to obtain a eutectic; S2: heating isopropyl stearate until it is completely melted to obtain pretreated isopropyl stearate; adding polyethylene glycol lecithin to deionized water, heating in a water bath to control the temperature, stirring to dissolve evenly, then slowly adding the pretreated isopropyl stearate and vitamin E, homogenizing to form a uniform emulsion, and continuously stirring until it is naturally cooled to room temperature to obtain a cooled emulsion; S3: adding the eutectic prepared in step S1 to anhydrous ethanol, stirring and dissolving uniformly to obtain a eutectic solution; then slowly adding the eutectic solution to the cooled emulsion prepared in step S2, stirring until the eutectic is uniformly distributed in the emulsion, and then slowly adding modified hydroxyethyl cellulose, stirring until the modified hydroxyethyl cellulose is completely dissolved, and adjusting the pH with sodium citrate and citric acid to obtain a dispersed emulsion; S4: After stirring the dispersed emulsion in step S3 at 500-1000 rpm for 10-30 min, the homogenization cycle is performed 3-5 times, the parameters are set to 300-500 bar, the temperature is room temperature, and each treatment is 2 min. After the homogenization treatment is completed, it is stirred again at 500-1000 rpm for 10 min to obtain an emulsified drug preparation.

3. The method for preparing a pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy according to claim 2, characterized in that: The ratio of the amount of febuxostat, p-aminobenzoic acid, salicylic acid and anhydrous ethanol used in step S1 is 1 mg:1.5 mg:1 mg:10 mL.

4. The method for preparing a pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy according to claim 3, characterized in that: The ratio of the amount of isopropyl stearate, polyethylene glycol lecithin, vitamin E and deionized water in step S2 is 5-10g: 1g: 50mL: 0.1-0.3g; the specific parameters of the homogenization treatment are an initial homogenization pressure of 500-800bar, a temperature of 40-60°C, a homogenization treatment time of 2-3min each time, and a total of 3 cycles. The homogenization pressure is increased to 1000-1200bar during the second homogenization treatment, and the other parameters are the same as the first treatment. The temperature is lowered to 20°C during the third homogenization treatment, and the other parameters are the same as the first treatment.

5. The method for preparing a pharmaceutical preparation for alleviating the progression of hyperuricemia nephropathy according to claim 4, characterized in that: The ratio of the amount of the eutectic, modified hydroxyethyl cellulose, cooling emulsion and anhydrous ethanol used in step S3 is 50 mg: 1-2 g: 100 mL: 50-100 mL.

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