Icodextrin preparation for peritoneal dialysis and preparation method thereof

Abstract

The invention relates to the technical field of pharmaceutical preparations, and particularly discloses an icodextrin preparation for peritoneal dialysis and a preparation method of the icodextrin preparation. According to the icodextrin preparation for peritoneal dialysis provided by the invention, the first independent solution and the second independent solution are respectively prepared by adopting a double-chamber structure, and the taurine, the quercetin and the hyperbranched polyglycerol are added into the first independent solution; the icodextrin peritoneal dialysis solution effectively solves the risks of metabolic disorder and peritoneal injury caused by a traditional glucose dialysis solution and peritoneal fibrosis and pipeline blockage existing in an existing icodextrin dialysis solution, the stability of the icodextrin peritoneal dialysis solution is remarkably improved, the probability of adverse reactions is reduced, and the icodextrin peritoneal dialysis solution is excellent in viscosity resistance and osmotic pressure stability and can be used for preparing peritoneal dialysis solutions. The peritoneal dialysis solution can well replace the existing peritoneal dialysis solution, is simple in preparation process and good in repeatability, is easy to realize industrial mass production, and has relatively high popularization and application values in peritoneal dialysis treatment of patients with end-stage nephropathy.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical preparation technology, and in particular to an icodextrin preparation for peritoneal dialysis and its preparation method. Background Technology

[0002] In the field of kidney replacement therapy (KRT), peritoneal dialysis (PD) is one of the important treatment methods for patients with end-stage renal disease. However, the traditional peritoneal dialysis fluids widely used in clinical practice currently use glucose as the core osmotic agent, which has many inherent limitations that are difficult to overcome. These limitations mainly manifest in the following ways: long-term use can easily induce volume imbalances in patients (such as fluid overload), a rapid decline in osmotic capacity, metabolic disorders (such as hyperglycemia, hyperinsulinemia, and lipid metabolism abnormalities), and progressive damage to peritoneal tissues, as well as the problem of increased viscosity during use. These inherent technical problems not only limit the long-term efficacy of PD but also affect its application in a wider range of patient populations.

[0003] To address the aforementioned shortcomings of traditional glucose peritoneal dialysis solutions, researchers have conducted extensive studies, with peritoneal dialysis solutions using icodextrin as an alternative osmotic agent becoming a hot research topic. Icodextrin, a large-molecule polysaccharide, is slowly metabolized in the body and can maintain effective osmotic pressure in the peritoneal cavity for a longer period, effectively solving problems such as elevated blood glucose, metabolic disorders, and volume overload caused by traditional glucose dialysis solutions. However, in clinical practice, peritoneal dialysis solutions using icodextrin as an osmotic agent also present some new technical problems, which seriously affect their clinical application efficacy. These problems include: First, long-term use of icodextrin peritoneal dialysis solutions can lead to pathological changes in the peritoneum in some patients, such as increased peritoneal inflammation and progression of peritoneal fibrosis, further impairing peritoneal function. Second, during peritoneal dialysis, the ultrafiltration performance of icodextrin gradually decreases with prolonged use, resulting in limited peritoneal filtration and failure to achieve the ideal dialysis treatment goals. Third, icodextrin solutions are prone to viscosity increases during storage and use, especially after mixing with other components or undergoing sterilization, which increases the risk of dialysis tubing blockage and affects the safety and stability of treatment.

[0004] Therefore, developing a peritoneal dialysis solution that can effectively solve the above problems while taking into account good dialysis effect, peritoneal protection performance and safety of use is of great significance for promoting the development of peritoneal dialysis technology and improving the treatment effect and quality of life of patients with end-stage renal disease. Summary of the Invention

[0005] In view of the problems existing in icodextrin peritoneal dialysis solutions, such as easy occurrence of peritoneal pathological changes, limited peritoneal filtration effect, and easy increase in viscosity during use or storage, the present invention provides an icodextrin preparation for peritoneal dialysis and its preparation method.

[0006] To solve the above-mentioned technical problems, the technical solution provided by the present invention is as follows:

[0007] An icodextrin preparation for peritoneal dialysis includes a first separate solution and a second separate solution; wherein the first separate solution includes icodextrin, taurine, quercetin, and hyperbranched polyglycerol; and the second separate solution includes an alkaline pH adjuster.

[0008] Compared to existing technologies, the icodextrin preparation for peritoneal dialysis provided by this invention uses icodextrin as the core permeabilizing agent in the first separate solution, with taurine added as a peritoneal protectant. This effectively reduces oxidative stress damage and peritoneal inflammation during peritoneal dialysis, minimizing side effects. Furthermore, taurine enhances dialysis efficacy, and its unique physicochemical properties help maintain a stable osmotic pressure gradient in the peritoneal cavity for a longer period, preventing rapid osmotic pressure drops that could lead to decreased ultrafiltration efficiency and ensuring the stability of long-term dialysis results. The addition of quercetin as a degradation inhibitor effectively reduces the decomposition rate of icodextrin, decreasing glucose degradation products (GDPs) generated by icodextrin degradation, while simultaneously blocking the formation and accumulation of advanced glycation end products (AGEs), thereby reducing peritoneal angiogenesis and peritoneal fibrosis. The addition of hyperbranched polyglycerol as an anti-sticking agent effectively reduces viscosity fluctuations in icodextrin solutions, mitigating viscosity increases caused by environmental temperature changes, prolonged storage time, or component interactions. This effectively reduces the risk of tubing blockage, ensures smooth dialysis procedures, and enhances the safety of clinical operations.

[0009] By storing the first separate solution and the second separate solution (alkaline pH adjuster) separately, icodextrin can be kept in a stable solution environment for a long time, effectively avoiding the risk of degradation caused by contact with alkaline substances, extending the shelf life of the preparation, ensuring the penetration efficiency and purity during clinical use, and ensuring the reliability of the therapeutic effect.

[0010] Furthermore, the molecular weight of the icodextrin raw material is 15000 Da to 19000 Da.

[0011] Furthermore, the mass ratio of icodextrin to taurine is 1:0.1 to 1:0.5.

[0012] The optimal ratio of taurine addition can enhance the oxidative stress clearance and anti-inflammatory effects of peritoneal dialysis fluid, while avoiding the metabolic burden or interference with peritoneal osmotic pressure caused by excessive taurine. Furthermore, the physicochemical properties of taurine can help icodextrin maintain osmotic pressure balance in the peritoneal cavity and prolong the osmotic pressure gradient stabilization time. In addition, the addition of taurine can better improve dialysis compliance and reliability, and reduce peritoneal sensitivity to dialysis fluid.

[0013] Furthermore, the mass ratio of icodextrin to quercetin is 1:0.005 to 1:0.01.

[0014] The optimal proportion of quercetin can significantly reduce the degradation rate of icodextrin during high-temperature sterilization. At the same time, quercetin can exert an antioxidant effect, reducing peritoneal stress and inflammatory responses induced by icodextrin.

[0015] Furthermore, the mass ratio of icodextrin to hyperbranched polyglycerol is 1:0.05 to 1:0.1.

[0016] Icodextrin solution is prone to viscosity increase due to molecular aggregation during storage and dialysis. The addition of a specific proportion of hyperbranched polyglycerol can, on the one hand, play a stable viscosity-reducing role, maintain the viscosity stability of the dialysate during the circulation in the peritoneal cavity, and reduce the risk of vascular blockage; on the other hand, it can provide a certain osmotic pressure, reducing the problem of decreased osmotic pressure in the later stages of dialysis.

[0017] Furthermore, the hyperbranched polyglycerol has a molecular weight of 8000 Da to 12000 Da.

[0018] Hyperbranched polyglycerol with a molecular size of 8000Da to 12000Da is similar to that of icodextrin and can be uniformly dispersed in icodextrin solutions, reducing icodextrin molecule aggregation and improving viscosity reduction. At the same time, hyperbranched polyglycerol with this molecular weight has suitable molecular flexibility and can form a continuous and thin lubricating film on the inner wall of dialysis tubing. When the molecular weight is too low (<8000Da), the lubricating film is easily washed away by water flow and cannot maintain the lubricating effect for a long time. When the molecular weight is too high (>12000Da), the molecular rigidity is enhanced, and the formed lubricating film is thicker, which may increase the frictional resistance of the inner wall of the tubing.

[0019] Furthermore, the first separate solution comprises icodextrin, taurine, quercetin, hyperbranched polyglycerol, sodium lactate, calcium chloride, and magnesium chloride, with a pH of 5.0–5.5.

[0020] Furthermore, the second separate solution comprises sodium bicarbonate, sodium hydroxide, and sodium chloride, with a pH of 6.5–7.5.

[0021] Before clinical use, the overall pH value of the dialysate can be adjusted to a neutral range (6.9-7.3) close to the physiological pH of the human body by mixing the first and second separate solutions within the specific pH range in a certain proportion. This avoids the stimulation of the peritoneum by a single acidic or alkaline solution, maintains the normal physiological function of peritoneal cells, improves the patient's treatment comfort, and reduces the occurrence of peritoneal intolerance reactions.

[0022] Furthermore, the first separate solution comprises the following components in parts by weight: 1 part icodextrin, 0.1 to 0.5 parts taurine, 0.005 to 0.01 parts quercetin, 0.05 to 0.1 parts hyperbranched polyglycerol, 0.05 to 0.07 parts sodium lactate, 0.002 to 0.004 parts calcium chloride, and 0.0005 to 0.0008 parts magnesium chloride; the second separate solution comprises the following components in parts by weight: 0.04 to 0.06 parts sodium bicarbonate and 0.02 to 0.04 parts sodium chloride.

[0023] Preferably, the first separate solution of the peritoneal dialysis icodextrin preparation comprises the following components in parts by weight: 1 part icodextrin, 0.3 parts taurine, 0.008 parts quercetin, 0.075 parts hyperbranched polyglycerol, 0.06 parts sodium lactate, 0.003 parts calcium chloride, and 0.0007 parts magnesium chloride; the second separate solution comprises the following components in parts by weight: 0.05 parts sodium bicarbonate and 0.03 parts sodium chloride.

[0024] Furthermore, the volume ratio of the first separate solution to the second separate solution is 1:1.5 to 1:3.

[0025] Specifically, the specifications of the icodextrin preparation for peritoneal dialysis described in this invention are 150g:2000mL / bag.

[0026] It should be noted that this invention employs a dual-chamber bag structure, consisting of two independent, non-communicating liquid chambers, used to hold the first and second separate solutions respectively. This ensures the stability of the components in the first and second separate solutions and prevents interactions. Icodextrin, under the stable conditions of the first separate solution, undergoes high-temperature sterilization, resulting in fewer sugar terminal products and glucose degradation products. This significantly reduces color changes and also minimizes side effects such as immunosuppression, pain, and fever caused by degradation products.

[0027] This invention constructs a stable icodextrin system using specific peritoneal protectants, degradation inhibitors, and anti-adhesion agents. This reduces the side effects of icodextrin on the body during long-term dialysis cycles, improves the stability of dialysis osmotic pressure, and reduces the content of degradation products. At the same time, the separate packaging of the first and second individual solutions helps to improve the stability of the formulation and ensure the safety and reliability of the treatment effect for patients.

[0028] This invention also provides a method for preparing an icodextrin preparation for peritoneal dialysis, comprising the following steps:

[0029] S1. Quercetin is dissolved in an alcohol solvent to obtain a quercetin solution; the quercetin solution is added to a polyvinyl alcohol aqueous solution, stirred and emulsified, the alcohol solvent is removed by rotary evaporation, ultrafiltration is performed, the residue is centrifuged, resuspended in water, concentrated, and freeze-dried to obtain quercetin nanoparticles.

[0030] S2, dissolve icodextrin, taurine, hyperbranched polyglycerol, sodium lactate, calcium chloride and magnesium chloride in 65% to 70% of the prescribed amount of water for injection, add the quercetin nanoparticles, mix evenly, adjust the pH to 5.0 to 5.5, add the remaining water for injection, filter, fill, sterilize, and obtain the first separate solution.

[0031] S3: Dissolve sodium bicarbonate and sodium chloride in 65%–70% of the prescribed amount of water for injection, adjust the pH to 7.4–8.5, add the remaining water for injection, filter, fill, sterilize, and obtain the second separate solution.

[0032] Quercetin itself is highly hydrophobic and poorly water-soluble. When added directly to the dialysate, it is easy to form local high-concentration aggregates due to insufficient dissolution. This not only affects its anti-degradation and anti-peritoneal fibrosis functions, but may also increase the risk of tubing blockage due to particle precipitation.

[0033] This invention first prepares quercetin into nanoparticles, ensuring uniform dispersion in the subsequent dialysate and avoiding uneven local concentrations due to low solubility. The prescribed amounts of icodextrin, taurine, hyperbranched polyglycerol, and electrolytes are dissolved in water for injection before the quercetin nanoparticles are added. This allows for uniform dispersion of quercetin in the icodextrin-electrolyte system, reducing the deviation in quercetin concentration per milliliter of dialysate and ensuring uniform distribution and sustained action of quercetin within the peritoneal cavity.

[0034] Preparing the first and second separate solutions separately allows for the formation of quercetin nanoparticles and hyperbranched polyglycerol protective layers on the icodextrin molecules, effectively reducing thermal collisions between icodextrin molecules and minimizing the formation of degradation products. Simultaneously, it ensures that icodextrin remains in a stable acidic environment during storage, preventing degradation caused by contact with sodium bicarbonate.

[0035] Furthermore, in S1, the alcohol solvent is anhydrous ethanol.

[0036] Furthermore, in S1, the concentration of the quercetin solution is 1 mg / mL to 3 mg / mL.

[0037] Further, in S1, the mass concentration of the polyvinyl alcohol aqueous solution is 0.2% to 1%, and its volume ratio with the alcohol solvent is 4:1 to 6:1.

[0038] Furthermore, in S1, the ultrafiltration uses an ultrafiltration membrane with a molecular weight cutoff of 10 kDa.

[0039] Specifically, in S1, the ultrafiltration, centrifugation, and resuspension steps are repeated 2 to 4 times to improve the purity of quercetin nanoparticles.

[0040] Specifically, in S1, the freeze-drying steps include: pre-freezing at -40℃ for 12h to 15h, then freeze-drying at -40℃ to -20℃ for 8h to 12h, raising the temperature to -20℃ to 0℃ and freeze-drying for 8h to 12h, and finally freeze-drying at -0℃ to 20℃ for 18h to 22h.

[0041] For example, the temperature of the water used for injection in the preparation process is 45°C to 49°C.

[0042] Furthermore, in S2, the pH is adjusted to 5.0–5.5 using a 0.1 mol / L hydrochloric acid solution; in S3, the pH is adjusted to 7.4–8.5 using a 1% sodium hydroxide solution.

[0043] Specifically, in S2 and S3, the filtration is carried out using a polyethersulfone filter element with a pore size of 0.45 μm.

[0044] Specifically, in S2 and S3, the sterilization is wet pressure sterilization, the sterilization temperature is 121°C, and the sterilization time is 15 minutes.

[0045] It should be noted that the preparation process of the first and second separate solutions shall not exceed 12 hours from the start to the end of solution preparation, and shall not exceed 24 hours from the start to the end of filling.

[0046] The icodextrin preparation for peritoneal dialysis provided by this invention employs a dual-chamber structure to prepare a first separate solution and a second separate solution. Taurine, quercetin, and hyperbranched polyglycerol are added to the first separate solution, effectively solving the metabolic disorders and peritoneal damage caused by traditional glucose dialysate, as well as the risks of peritoneal fibrosis and tubing blockage present in existing icodextrin dialysate. It significantly improves the stability of icodextrin peritoneal dialysis solution, reduces the probability of adverse reactions, and exhibits excellent anti-adhesion and osmotic pressure stability. It can be a good substitute for existing peritoneal dialysis solutions. Moreover, the preparation process is simple, reproducible, and easy to achieve large-scale industrial production, making it highly valuable for promotion and application in peritoneal dialysis treatment of end-stage renal disease patients. Detailed Implementation

[0047] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0048] To better illustrate the present invention, further examples are provided below.

[0049] The molecular weight of the icodextrin used in the following examples is 15,000 Da to 19,000 Da. The molecular weight of the hyperbranched polyglycerol is 8,000 Da to 12,000 Da.

[0050] The original reagent was Batch numbers G23052226 and G24032726.

[0051] Example 1

[0052] This embodiment provides an icodextrin preparation for dialysis (specification: 150g:2000mL / bag). The formula for preparing 500 bags of the preparation is shown in the table below:

[0053]

[0054] The preparation method of the above-mentioned icodextrin preparation for dialysis is as follows:

[0055] S1, add the prescribed amount of quercetin to anhydrous ethanol to obtain a quercetin solution with a concentration of 2 mg / mL; under 250W probe ultrasound, add the quercetin solution dropwise to 5 times the volume of anhydrous 0.5 wt% polyvinyl alcohol aqueous solution, stir and emulsify for 24 h, remove ethanol by rotary evaporation, perform ultrafiltration using an ultrafiltration membrane with a molecular weight cutoff of 10 kD, centrifuge the residue at 10000 g for 30 min, resuspend in deionized water, repeat the above ultrafiltration, centrifugation, and resuspension steps three times, finally concentrate, pre-freeze the concentrate at -40℃ for 14 h, then freeze-dry at -30℃ for 10 h, raise the temperature to -10℃, freeze-dry for 10 h, and finally freeze-dry at -5℃ for 20 h to obtain quercetin nanoparticles;

[0056] S2, dissolve the prescribed amounts of icodextrin, taurine, hyperbranched polyglycerol, sodium lactate, calcium chloride, and magnesium chloride in 68% of the prescribed amount of water for injection at a temperature of 47°C. Add the above-mentioned quercetin nanoparticles, stir for 45 min, adjust the pH to 5.3 with 0.1 mol / L hydrochloric acid solution, add the remaining water for injection, filter through a 0.45 μm polyethersulfone filter, fill into A bags with a volume of 840 mL, seal, and sterilize at 121°C for 15 min to obtain the A bag solution;

[0057] S3, dissolve the prescribed amount of sodium bicarbonate and sodium chloride in 68% of the prescribed amount of water for injection at a temperature of 47°C, adjust the pH to 7.8 with a 1% sodium hydroxide solution, add the remaining water for injection, filter through a 0.45μm polyethersulfone filter, fill into B bags with a volume of 1260mL, seal, and sterilize at 121°C for 15min to obtain the B bag solution.

[0058] Example 2

[0059] This embodiment provides an icodextrin preparation for dialysis (specification: 150g:2000mL / bag). The formula for preparing 100 bags of the preparation is shown in the table below:

[0060]

[0061]

[0062] The preparation method of the above-mentioned icodextrin preparation for dialysis is as follows:

[0063] S1, add the prescribed amount of quercetin to anhydrous ethanol to obtain a quercetin solution with a concentration of 1 mg / mL; under 250W probe ultrasound, add the quercetin solution dropwise to 4 times the volume of anhydrous 0.2 wt% polyvinyl alcohol aqueous solution, stir and emulsify for 24 h, remove ethanol by rotary evaporation, perform ultrafiltration using an ultrafiltration membrane with a molecular weight cutoff of 10 kD, centrifuge the residue at 10000 g for 30 min, resuspend in deionized water, repeat the above ultrafiltration, centrifugation, and resuspension steps three times, finally concentrate, pre-freeze the concentrate at -40℃ for 12 h, then freeze-dry at -35℃ for 8 h, raise the temperature to -20℃, freeze-dry for 8 h, and finally freeze-dry at 0℃ for 18 h to obtain quercetin nanoparticles;

[0064] S2, dissolve the prescribed amounts of icodextrin, taurine, hyperbranched polyglycerol, sodium lactate, calcium chloride, and magnesium chloride in 65% of the prescribed amount of water for injection at a temperature of 45°C. Add the above-mentioned quercetin nanoparticles, stir for 45 min, adjust the pH to 5.5 with 0.1 mol / L hydrochloric acid solution, add the remaining water for injection, filter through a 0.45 μm polyethersulfone filter, fill into A bags with a volume of 840 mL, seal, and sterilize at 121°C for 15 min to obtain the A bag solution;

[0065] S3, dissolve the prescribed amount of sodium bicarbonate and sodium chloride in 65% of the prescribed amount of water for injection at a temperature of 45°C, adjust the pH to 7.4 with a 1% sodium hydroxide solution, add the remaining water for injection, filter with a 0.45μm polyethersulfone filter, fill into B bags with a volume of 1260mL, seal, and sterilize at 121°C for 15min to obtain the B bag solution.

[0066] Example 3

[0067] This embodiment provides an icodextrin preparation for dialysis (specification: 150g:2000mL / bag). The formula for preparing 100 bags of the preparation is shown in the table below:

[0068]

[0069]

[0070] The preparation method of the above-mentioned icodextrin preparation for dialysis is as follows:

[0071] S1, add the prescribed amount of quercetin to anhydrous ethanol to obtain a quercetin solution with a concentration of 3 mg / mL; under 250W probe ultrasound, add the quercetin solution dropwise to 6 times the volume of anhydrous 1wt% polyvinyl alcohol aqueous solution, stir and emulsify for 24 h, remove ethanol by rotary evaporation, perform ultrafiltration using an ultrafiltration membrane with a molecular weight cutoff of 10 kD, centrifuge the residue at 10000g for 30 min, resuspend in deionized water, repeat the above ultrafiltration, centrifugation, and resuspension steps three times, finally concentrate, pre-freeze the concentrate at -40℃ for 15 h, then freeze-dry at -20℃ for 12 h, raise the temperature to 0℃, freeze-dry for 12 h, and finally freeze-dry at 20℃ for 22 h to obtain quercetin nanoparticles;

[0072] S2, dissolve the prescribed amounts of icodextrin, taurine, hyperbranched polyglycerol, sodium lactate, calcium chloride, and magnesium chloride in 70% of the prescribed amount of water for injection at a temperature of 49°C. Add the above-mentioned quercetin nanoparticles, stir for 45 min, adjust the pH to 5.0 with 0.1 mol / L hydrochloric acid solution, add the remaining water for injection, filter through a 0.45 μm polyethersulfone filter, fill into A bags with a volume of 840 mL, seal, and sterilize at 121°C for 15 min to obtain the A bag solution;

[0073] S3, dissolve the prescribed amount of sodium bicarbonate and sodium chloride in 70% of the prescribed amount of water for injection at a temperature of 49°C, adjust the pH to 8.5 with a 1% sodium hydroxide solution, add the remaining water for injection, filter with a 0.45μm polyethersulfone filter, fill into B bags with a volume of 1260mL, seal, and sterilize at 121°C for 15min to obtain the B bag solution.

[0074] Comparative Example 1

[0075] This comparative example is an icodextrin preparation for dialysis (specification: 150g:2000mL / bag), which differs from Example 1 only in that it does not contain taurine. The formula for preparing 100 bags of the preparation is shown in the table below:

[0076]

[0077] The preparation method of the above-mentioned dialysis icorubicin preparation is the same as that in Example 1 except that taurine is omitted in the preparation process. The rest is exactly the same and will not be described again here.

[0078] Comparative Example 2

[0079] This comparative example is an icodextrin preparation for dialysis (specification: 150g: 2000mL / bag), which differs from Example 1 only in that it does not contain quercetin. The formulation for preparing 100 bags of the preparation is shown in the table below:

[0080]

[0081]

[0082] The preparation method of the above-mentioned dialysis icorubicin preparation differs from that in Example 1 only in that quercetin is omitted during the preparation process; otherwise, they are completely the same and will not be described again here.

[0083] Comparative Example 3

[0084] This comparative example is an icodextrin preparation for dialysis (specification: 150g: 2000mL / bag), which differs from Example 1 only in that it does not contain hyperbranched polyglycerol. The formulation for preparing 100 bags of the preparation is shown in the table below:

[0085]

[0086] The preparation method of the above-mentioned dialysis icorubicin preparation differs from that in Example 1 only in that hyperbranched polyglycerol is omitted during the preparation process; otherwise, they are completely the same and will not be described again here.

[0087] Comparative Example 4

[0088] This comparative example is an icodextrin preparation for dialysis (specification: 150g:2000mL / bag), which differs from Example 1 only in that taurine is replaced with an equal amount of glutathione. The formulation for preparing 100 bags of the preparation is shown in the table below:

[0089]

[0090] The preparation method of the above-mentioned icodextrin preparation for dialysis is the same as that in Example 1, and will not be repeated here.

[0091] Comparative Example 5

[0092] This comparative example is an icodextrin preparation for dialysis (specification: 150g:2000mL / bag), which differs from Example 1 only in that quercetin is replaced with an equal amount of sodium metabisulfite. The formulation for preparing 100 bags of the preparation is shown in the table below:

[0093]

[0094]

[0095] The preparation method of the above-mentioned icodextrin preparation for dialysis differs from that in Example 1 only in that sodium pyrosulfate is added directly without the preparation of nanoparticles; otherwise, they are completely the same and will not be described again here.

[0096] Comparative Example 6

[0097] This comparative example is an icodextrin preparation for dialysis (specification: 150g:2000mL / bag), which differs from Example 1 only in that hyperbranched polyglycerol is replaced with an equal amount of PEG 400. The formulation for preparing 100 bags of the preparation is shown in the table below:

[0098]

[0099]

[0100] The preparation method of the above-mentioned icodextrin preparation for dialysis is the same as that in Example 1, and will not be repeated here.

[0101] Comparative Example 7

[0102] This comparative example is an icodextrin preparation for dialysis (specification: 150g:2000mL / bag). The only difference from Example 1 is that the solutions in bag A and bag B are combined, packaged, and sterilized. The formulation for preparing 100 bags is exactly the same as in Example 1, and will not be repeated here. The preparation method is as follows:

[0103] S1, add the prescribed amount of quercetin to anhydrous ethanol to obtain a quercetin solution with a concentration of 2 mg / mL; under 250W probe ultrasound, add the quercetin solution dropwise to 5 times the volume of anhydrous 0.5 wt% polyvinyl alcohol aqueous solution, stir and emulsify for 24 h, remove ethanol by rotary evaporation, perform ultrafiltration using an ultrafiltration membrane with a molecular weight cutoff of 10 kD, centrifuge the residue at 10000 g for 30 min, resuspend in deionized water, repeat the above ultrafiltration, centrifugation, and resuspension steps three times, finally concentrate, pre-freeze the concentrate at -40℃ for 14 h, then freeze-dry at -30℃ for 10 h, raise the temperature to -10℃, freeze-dry for 10 h, and finally freeze-dry at -5℃ for 20 h to obtain quercetin nanoparticles;

[0104] S2, dissolve the prescribed amounts of icodextrin, taurine, hyperbranched polyglycerol, sodium lactate, calcium chloride, and magnesium chloride in 68% of the prescribed amount of water for injection at a temperature of 47°C, add the above-mentioned quercetin nanoparticles, stir for 45 min, adjust the pH to 5.3 with 0.1 mol / L hydrochloric acid solution, add the remaining water for injection, filter through a 0.45 μm polyethersulfone filter, and fill into bag A to obtain bag A solution;

[0105] S3, dissolve the prescribed amount of sodium bicarbonate and sodium chloride in 68% of the prescribed amount of water for injection at a temperature of 47°C, adjust the pH to 7.0 with a 1% sodium hydroxide solution, add the remaining water for injection, and filter with a 0.45μm polyethersulfone filter to obtain the B bag solution;

[0106] S4. Mix 840mL of solution from bag A and 1260mL of solution from bag B, fill the package, seal the package, and sterilize at 121℃ for 15 minutes to obtain icodextrin preparation for dialysis.

[0107] Quality Research

[0108] 1. Content of icodextrin and 5-hydroxymethylfurfural

[0109] 5-Hydroxymethylfurfural (HMF) is a degradation product of icodextrin, one of the controlled impurities in icodextrin preparations, and also one of the toxic substances that can cause adverse reactions. An influencing factor experiment was conducted on the icodextrin preparations for dialysis prepared in Example 1, Comparative Examples 2, 5, and 7. The icodextrin content and 5-hydroxymethylfurfural content were determined according to the sampling time points, and the results are shown in Table 2.

[0110] The content of icodextrin was determined according to the optical rotation determination method (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0621).

[0111] Test solution: Accurately measure 20 mL of this product and place it in a 100 mL volumetric flask. Dilute with water to the mark and shake well to obtain the test solution.

[0112] Measurement conditions: Concentration: 1.5 g / 100 mL, Temperature: 20℃, Number of measurements: 3, Test tube length: 10 cm. (A blank calibration with solvent should be performed before each measurement, and a second calibration should be performed after each measurement to determine if the zero point has changed during the measurement; if the difference in optical rotation exceeds +0.01 during the second calibration, it indicates that the zero point has changed, and the optical rotation should be measured again.)

[0113] Determination method: Rinse the test tube several times with the test solution, slowly inject an appropriate amount of the test solution (be careful not to generate air bubbles), place it in a polarimeter, measure the optical rotation, and calculate the icodextrin content according to the following formula.

[0114] Icodextrin (%) = [(optical rotation × 100 × 100 ÷ 20) - (-12.501 × L × C] 乳酸 Sodium) ÷ (195 × L × 7.5) × 100%

[0115] In the formula, L is the length of the optical rotation tube (dm); -12.501 is the specific rotation of the raw sodium lactate solution; C 乳酸钠The concentration of sodium lactate in the sample (g / 100ml) was determined by the sodium lactate method under the content determination section; 195 is the median of the specific rotation limit range for the Icodextrin raw material;

[0116] Sodium lactate was tested by high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512).

[0117] The test solution is obtained by taking this product.

[0118] Reference solution: Weigh approximately 45 mg of sodium lactate reference standard accurately, place it in a 10 mL volumetric flask, dissolve and dilute with water to the mark, shake well, and prepare a solution containing 4.5 mg of sodium lactate per 1 mL.

[0119] Chromatographic conditions

[0120] Column: Ultimate AQ C 18 4.6×250mm, 5μm;

[0121] The mobile phase consisted of 0.1% formic acid and 0.1% dicyclohexylamine.

[0122] The detection wavelength is 210 nm;

[0123] The column temperature is 30℃;

[0124] The flow rate was 1.0 mL / min;

[0125] The injection volume was 20 μL;

[0126] Running time: 20 minutes.

[0127] The test solution and reference solution were precisely measured and injected into the liquid chromatograph separately. The chromatograms were recorded, and the sodium lactate content was calculated by peak area using the external standard method.

[0128] Calculation formula

[0129] Where: f: correction factor; m 对 : Sample weight of reference standard (mg); P: Sodium lactate reference standard content (%);

[0130] A 对 Peak area of ​​the reference solution; A 供 Peak area of ​​the test solution.

[0131] Determination of 5-hydroxymethylfurfural: High performance liquid chromatography was used for detection.

[0132] Chromatographic conditions: Column: GL Sciences Inertsustain C 18(250mm×4.6mm, 5μm) Mobile phase A was 0.01mol / L sodium heptanesulfonate solution (containing 0.1% triethylamine, pH adjusted to 3.0 with phosphoric acid), mobile phase B was acetonitrile, gradient elution was performed according to Table 1, column temperature was 30℃, flow rate was 1.0mL / min, injection volume was 10μL, and detection wavelength was 284nm.

[0133] Table 1 Gradient elution program for mobile phase

[0134] Time (min) Mobile phase A (%) Mobile phase B (%) 0 85 15 8 85 15 8.1 50 50 16 50 50 16.1 85 85 30 85 85

[0135] Particle size distribution was determined using a laser particle size analyzer: Icodextrin formulations were taken and measured three times using a laser particle size analyzer, and the particle size distribution was recorded. The test results are shown in Table 2.

[0136] Table 2. Experimental results of influencing factors in the examples and comparative examples.

[0137]

[0138] The results showed that, in Comparative Example 2, which did not contain quercetin, the content of 5-hydroxymethylfurfural increased significantly with prolonged storage time. In Comparative Example 5, glutathione was used to replace quercetin, but its inhibitory effect on degradation products was not as good as that of quercetin. In Comparative Example 7, where A and B bags were combined before sterilization, a significant increase in 5-hydroxymethylfurfural was observed.

[0139] 2. Viscosity test

[0140] Icodextrin dialysates prepared in Examples 1, the original drug, Comparative Examples 3 and 6 were used in a clinical dialysis system for 8 hours after being inserted into a 4800 mL blood bank. Samples were then collected for later use. For measurement, samples of the prepared icodextrin dialysates were transferred, and the viscosity of the corresponding samples was determined using the capillary method while maintaining a constant temperature. The outflow time t of the solution was measured using an Ubbelohde viscometer, and the measurement was repeated three times using a stopwatch (minimum unit 1 / 100 s) (error less than 0.2 s). The average value of the three measurements was taken. The outflow time t0 of the solvent water was measured using the same method. The relative viscosity value η was calculated using the following formula. r The results are shown in Table 3.

[0141]

[0142] The original manufacturer is Guangzhou Baxter Medical Supplies Co., Ltd., and the specification is 7.5% icodextrin dialysis solution, batch number G23052226.

[0143] Table 3 Viscosity measurement results (n=3)

[0144]

[0145]

[0146] 3. Peritoneal dialysis capacity study

[0147] Eight-week-old male Sprague-Dawley rats were housed in an SPF-grade animal facility with a 12-hour light-dark cycle and provided with a standard rodent diet. The original control group received intraperitoneal injections of 2 mL / 100 g / day of the original dialysis solution for four consecutive weeks. The original solution was manufactured by Guangzhou Baxter Medical Supplies Co., Ltd., and its specification was 7.5% icodextrin dialysis solution, batch number G23052226.

[0148] Peritoneal dialysis Example 1 group: 2 mL / 100 g·d of Example 1 peritoneal dialysis solution was injected intraperitoneally for 4 consecutive weeks; Peritoneal dialysis Comparative Example 1 group: 2 mL / 100 g·d of Comparative Example 1 peritoneal dialysis solution was injected intraperitoneally for 4 consecutive weeks; Peritoneal dialysis Comparative Example 2 group: 2 mL / 100 g·d of Comparative Example 2 peritoneal dialysis solution was injected intraperitoneally for 4 consecutive weeks; Peritoneal dialysis Comparative Example 3 group: 2 mL / 100 g·d of Comparative Example 3 peritoneal dialysis solution was injected intraperitoneally for 4 consecutive weeks; Peritoneal dialysis Comparative Example 4 group: 2 mL / 100 g·d of Comparative Example 4 peritoneal dialysis solution was injected intraperitoneally for 4 consecutive weeks.

[0149] Thirty minutes before sacrifice, 2 mL of 4.25% glucose peritoneal dialysis fluid was slowly injected into the peritoneal cavity of the rats, taking care to avoid damaging the abdominal organs during the injection. Dialysis fluid samples were collected at 30 minutes and 0 minutes after the start of peritoneal dialysis (i.e., before the injection of dialysis fluid) to determine the concentrations of glucose and blood urea nitrogen (BUN) in the fluid.

[0150] Glucose transport rate = C 30 (Glucose concentration in dialysate at 30 min) / C0 (Glucose concentration in dialysate at 0 min) × 100%.

[0151] Urea nitrogen transport rate = CD (BUN concentration in dialysate at 30 min) / CP (BUN concentration in blood at 0 min) × 100%.

[0152] In the dialysis experiment, the smaller the thickness of the peritoneal dialysis membrane, the less impact dialysis has on the peritoneum; the higher the glucose transport rate, the better the peritoneum's glucose filtration capacity, and the smaller the impact of the dialysate on the peritoneum; the higher the urea nitrogen transport rate, the more the peritoneum is in a high transport state, and the greater the impact of the dialysate on the peritoneum. The results are shown in Table 4.

[0153] Peritoneal thickness was measured using peritoneal tissue from rats that had been on dialysis for 4 weeks. The tissue was stained with H&E and then the thickness was measured using a microscope. The results are shown in Table 4.

[0154] Table 4. Results of peritoneal balance test in rats after dialysis, detecting changes in peritoneal function.

[0155]

[0156] The above experiments show that, compared with the original drug, Example 1 exhibits significant advantages in peritoneal thickness (71.01±2.30μm), glucose transport efficiency (0.41±0.03%), and urea nitrogen transport efficiency (0.56±0.01%). Peritoneal thickness did not increase significantly, and glucose transport efficiency was superior to the original drug (0.35±0.21%) and most comparative examples, demonstrating its more efficient solute clearance capacity. This indicates that taurine shows certain advantages in stabilizing the peritoneum and glucose transport efficiency. The urea nitrogen transport efficiency was 8.2% lower than the original drug membrane (0.61±0.02%), and superior to most comparative examples, indicating a smaller impact on the peritoneum. Compared to Example 1, Comparative Example 1 showed a significant increase in peritoneal thickness, a decrease in glucose transport rate, and an increase in urea nitrogen transport rate, further demonstrating the superiority and safety of taurine in this invention. Compared to Example 1, Comparative Examples 2 and 3 showed no significant changes in peritoneal thickness, glucose transport rate, and urea nitrogen transport rate, indicating that these indicators are related to taurine but not significantly related to quercetin or hyperbranched polyglycerol. In Comparative Example 4, replacing taurine with glutathione did not show the same significant advantages in membrane thickness, glucose transport rate, and urea transport rate as Example 1. This demonstrates that the taurine introduced in this invention has unexpected effects. These data prove that this invention represents a breakthrough improvement in dialysis efficiency and biocompatibility, making it suitable for long-term peritoneal dialysis applications.

[0157] 4. Studies on peritoneal fibrosis and inflammation:

[0158] Eight-week-old male SD rats were housed in an SPF-grade animal facility under 12-hour light-dark cycles and provided with a standard rodent diet. The rats were randomly divided into five groups (n=6 per group): the control group received intraperitoneal injection of 0.9% NaCl aqueous solution (10 mL / 100 g·d), while the other five groups received injections of Example 1, the original formulation, and, respectively, icodextrin peritoneal dialysis solution (2 mL / 100 g·d) from Comparative Examples 1, 2, and 4, for four consecutive weeks. After the experiment, peritoneal tissue was minced and weighed (approximately 100 mg), and homogenized in RIPA lysis buffer (1 mL) containing 1 mM PMSF. After centrifugation at 12000 rpm for 10 min, the supernatant was collected as the total protein extract. The protein sample was mixed with 5×SDS-PAGE loading buffer at a 1:4 volume ratio and denatured in a boiling water bath for 10 min.

[0159] Follow the instructions of each kit, cool the sample and load it (5-10 μL / well). After electrophoresis on a stacking gel (80V, 30 min) and a separating gel (120V, 60 min), transfer the sample to a membrane (α-SMA 45 min, Beclin-1 53 min, LC3B 20 min, P62 50 min). Blocking was performed using 5% skim milk powder at room temperature for 2 hours, followed by incubation overnight at 4°C with primary antibodies (the names and dilution ratios of the primary antibodies were Alpha-Smooth Muscle Actin (α-SMA) (Aladdin, catalog number: Ab088443) 1:1500, Beclin-1 (Abcam, catalog number: ab232461) 1:1500, Microtubule-Associated Protein 1A / 1B-Light Chain3B (LC3B) (Abcam, catalog number: ab222776) 1:1000, and Sequestosome-1 (P62) (Abcam, catalog number: ab222776) 1:1000, v / v). After washing three times with PBST, HRP-labeled secondary antibody (1:20000) (Thermo Fisher, catalog number: SA1-300) was added and incubated at room temperature for 2 hours. After washing again, the protein was developed, and the expression of the target protein was quantitatively analyzed by band density. The results are shown in Table 5.

[0160] Table 5. Results of peritoneal inflammatory protein expression in each group of rats

[0161] Group Beclin-1 LC3-Ⅱ LC3-Ⅰ P62 A-SMA Original research group 0.19±0.06 0.14±0.07 0.19±0.07 0.53±0.07 0.23±0.08 Example 1 0.12±0.05* 0.08±0.05* 0.10±0.05* 0.64±0.05* 0.12±0.06** Comparative Example 1 0.59±0.08** 0.31±0.10** 0.41±0.07** 0.28±0.09** 1.20±0.23** Comparative Example 2 0.23±0.06** 0.19±0.09 0.25±0.04 0.49±0.12 0.29±0.07 Comparative Example 4 0.21±0.06** 0.16±0.08 0.17±0.05 0.64±0.05 0.16±0.06*

[0162] Note: Compared with the original research group, *p<0.05, **p<0.01

[0163] The results showed that Western blot analysis of autophagy-related proteins (Beclin-1, LC3-II, LC3-I, P62) and fibrosis markers (α-SMA) after peritoneal dialysis fluid treatment revealed significant differences in the effects of different treatment groups on peritoneal autophagy and fibrosis progression. Compared with the original study group, the Example 1 group exhibited the most significant anti-fibrotic effect, with significantly reduced expression of Beclin-1 (0.12±0.05*), LC3-II (0.08±0.05*), and LC3-I (0.10±0.05), while increased expression of P62 (0.64±0.05) and significantly downregulated α-SMA (0.12±0.06), suggesting that this group may have alleviated peritoneal fibrosis by inhibiting excessive autophagy. In contrast, Comparative Example 1 exhibited characteristics of excessive autophagy activation, with significantly elevated Beclin-1 (0.59±0.08), LC3-II (0.31±0.10), and LC3-I (0.41±0.07), and decreased P62 (0.28±0.09), accompanied by an abnormally high α-SMA (1.20±0.23), suggesting that autophagy disorder may exacerbate the fibrosis process. Comparative Example 2 showed only an increase in Beclin-1 (0.23±0.06) without significant changes in other indicators. While Comparative Example 4 showed an increase in Beclin-1 (0.21±0.06) and a decrease in α-SMA (0.16±0.06*), the effects were weaker than those in Example 1.

[0164] The overall results indicate that the peritoneal dialysis fluid of Example 1 has the best effect in regulating autophagy balance and inhibiting peritoneal fibrosis, while Comparative Example 1 may promote fibrosis due to abnormal autophagy. This provides an important basis for subsequent optimization of peritoneal dialysis fluid formulation.

[0165] 5. Measurement of inflammatory factors

[0166] Peritoneal tissue sections from each group were treated with permeabilization solution for 30 min; blocked with goat serum for 30 min; primary antibodies (IL-6, IL-10, TNF-α) were added, and incubated at 37℃ for 1 h; washed three times with PBS, and fluorescent secondary antibody (1:400) was added, and incubated at 37℃ in the dark for 30 min; after washing with PBS, cell nuclei were counterstained with DAPI for 5 min; rinsed with PBS, and mounted with anti-fluorescence quenching mounting medium; images were acquired using a fluorescence microscope, and screenshots were taken using CaseViewer software. The mean optical density (MOD) of the green fluorescence signal was quantitatively analyzed using the JD801 medical image analysis system. The results are shown in Table 6.

[0167] Table 6. Data on the levels of inflammatory factors in different groups of rat peritoneal tissue.

[0168]

[0169]

[0170] The results showed that this study systematically evaluated the anti-inflammatory effects of different peritoneal dialysis solutions by detecting the expression levels of key inflammatory factors (IL-6, IL-10, and TNF-α). Compared with the original group (IL-6 0.24±0.06, IL-10 0.35±0.12, TNF-α 0.24±0.10), the Example 1 group exhibited the best anti-inflammatory properties. Its pro-inflammatory factors IL-6 (0.17±0.09) and TNF-α (0.18±0.08) were significantly reduced, while the anti-inflammatory factor IL-10 (0.45±0.15) was significantly increased, forming a good anti-inflammatory-pro-inflammatory balance. In contrast, Comparative Example 1 showed a significant pro-inflammatory tendency, with IL-6 (0.39±0.12) and TNF-α (0.46±0.07) increasing by 62.5% and 91.7%, respectively, while IL-10 (0.31±0.10) decreased by 11.4%, suggesting that the formulation may induce an inflammatory response. Comparative Example 2 and Comparative Example 4 showed improvements in IL-6 levels (0.28±0.14 and 0.27±0.09, respectively), but their regulatory effects on TNF-α and IL-10 were limited, and their anti-inflammatory effects were far inferior to those of Example 1. It is noteworthy that Example 1 demonstrated a significant advantage in inflammation regulation and exhibited multi-target protective effects against peritoneal hyperplasia and fibrosis, making it the most promising candidate formulation for clinical application. This further demonstrates the advantages of the Example 1 formulation in anti-inflammatory, anti-proliferative, and anti-fibrotic effects.

[0171] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions or improvements made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. An icodextrin preparation for peritoneal dialysis, characterized in that, It includes a first separate solution and a second separate solution; wherein the first separate solution includes icodextrin, taurine, quercetin and hyperbranched polyglycerol; and the second separate solution includes an alkaline pH adjuster.

2. The icodextrin preparation for peritoneal dialysis as described in claim 1, characterized in that, The mass ratio of icodextrin to taurine is 1:0.1 to 1:0.5; and / or The mass ratio of icodextrin to quercetin is 1:0.005 to 1:0.01; and / or The mass ratio of icodextrin to hyperbranched polyglycerol is 1:0.05 to 1:0.

1.

3. The icodextrin preparation for peritoneal dialysis as described in claim 1 or 2, characterized in that, The hyperbranched polyglycerol has a molecular weight of 8000 Da to 12000 Da.

4. The icodextrin preparation for peritoneal dialysis as described in claim 1 or 2, characterized in that, The first separate solution comprises icodextrin, taurine, quercetin, hyperbranched polyglycerol, sodium lactate, calcium chloride, and magnesium chloride, with a pH of 5.0–5.5; and / or The second separate solution comprises sodium bicarbonate, sodium hydroxide, and sodium chloride, with a pH of 6.5–7.

5.

5. The icodextrin preparation for peritoneal dialysis as described in claim 4, characterized in that, The first separate solution comprises the following components in parts by weight: 1 part icodextrin, 0.1 to 0.5 parts taurine, 0.005 to 0.01 parts quercetin, 0.05 to 0.1 parts hyperbranched polyglycerol, 0.05 to 0.07 parts sodium lactate, 0.002 to 0.004 parts calcium chloride, and 0.0005 to 0.0008 parts magnesium chloride; The second separate solution comprises the following components in parts by mass: 0.04 to 0.06 parts sodium bicarbonate and 0.02 to 0.04 parts sodium chloride.

6. The icodextrin preparation for peritoneal dialysis as described in claim 5, characterized in that, The first separate solution comprises the following components in parts by mass: 1 part icodextrin, 0.3 parts taurine, 0.008 parts quercetin, 0.075 parts hyperbranched polyglycerol, 0.06 parts sodium lactate, 0.003 parts calcium chloride, and 0.0007 parts magnesium chloride; The second separate solution comprises the following components in parts by mass: 0.05 parts sodium bicarbonate and 0.03 parts sodium chloride.

7. The icodextrin preparation for peritoneal dialysis as described in claim 1, characterized in that, The volume ratio of the first separate solution to the second separate solution is 1:1.5 to 1:

3.

8. A method for preparing an icodextrin preparation for peritoneal dialysis, characterized in that, Includes the following steps: S1, Quercetin is dissolved in an alcohol solvent to obtain a quercetin solution; the quercetin solution is added to a polyvinyl alcohol aqueous solution, stirred and emulsified, the alcohol solvent is removed by rotary evaporation, ultrafiltration is performed, the residue is centrifuged, resuspended in water, concentrated, and freeze-dried to obtain quercetin nanoparticles. S2, dissolve icodextrin, taurine, hyperbranched polyglycerol, sodium lactate, calcium chloride and magnesium chloride in 65% to 70% of the prescribed amount of water for injection, add the quercetin nanoparticles, mix evenly, adjust the pH to 5.0 to 5.5, add the remaining water for injection, filter, fill, sterilize, and obtain the first separate solution. S3: Dissolve sodium bicarbonate and sodium chloride in 65%–70% of the prescribed amount of water for injection, adjust the pH to 7.4–8.5, add the remaining water for injection, filter, fill, sterilize, and obtain the second separate solution.

9. The method for preparing the icodextrin preparation for peritoneal dialysis as described in claim 8, characterized in that, In S1, the alcohol solvent is anhydrous ethanol; and / or In S1, the concentration of the quercetin solution is 1 mg / mL to 3 mg / mL; and / or In S1, the mass concentration of the polyvinyl alcohol aqueous solution is 0.2% to 1%, and its volume ratio with the alcohol solvent is 4:1 to 6:

1.

10. The method for preparing the icodextrin preparation for peritoneal dialysis as described in claim 8, characterized in that, In S1, the ultrafiltration uses an ultrafiltration membrane with a molecular weight cutoff of 10 kDa.

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