Special blood filtration replacement liquid for anticoagulation

Abstract

The invention discloses blood filtration replacement liquid as well as a preparation method and application thereof. The displacement liquid comprises low molecular weight heparin calcium, prostaglandin E1, prostacyclin, electrolyte with specific concentration, glucose and an interleukin-1 receptor antagonist. The preparation method comprises the following steps: weighing the raw materials according to the proportion, dissolving to a constant volume, removing pyrogen and heat, and carrying out aseptic packaging. During clinical application, replacement liquid and blood are filtered according to the replacement rate of 1: 1-2: 1 according to the condition of a patient, 4-6 hours each time and 2-3 times each week, and related indexes are monitored. Animal tests show that the replacement liquid has good safety under the effective dosage, and has potential influence on part of organs under the high dosage. A stability test shows that the compound can be stably stored under a certain condition. The invention provides the replacement liquid which can achieve anticoagulation and blood vessel protection, maintain electrolyte balance and relieve inflammatory response, is expected to improve the prognosis of hemodialysis patients, and has important clinical significance and wide application prospects.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical and food technology, and in particular to a blood filtration replacement solution specifically for anticoagulation. Background Technology

[0002] Hemodialysis is one of the essential renal replacement therapies for patients with end-stage renal disease (ESRD). With the increasing aging of the global population and the rising incidence of chronic diseases, the number of ESRD patients is increasing year by year, leading to a growing demand for hemodialysis treatment. Statistics show that in my country alone, a large number of newly diagnosed ESRD patients require hemodialysis treatment each year.

[0003] Hemodialysis is a technique that involves drawing a patient's blood out of the body, filtering and purifying it through a dialyzer to remove accumulated metabolic waste, excess water, and toxins. The purified blood is then returned to the patient, partially replacing kidney function and maintaining the patient's vital functions and internal homeostasis. Despite continuous development and improvement, this technology still faces numerous challenges and unresolved issues.

[0004] Traditional hemodialysis replacement fluids primarily rely on single anticoagulants, such as heparin or low molecular weight heparin, for anticoagulation. While these anticoagulants can prevent blood clotting during dialysis to some extent, they often come with a high risk of bleeding. Heparin's anticoagulant mechanism involves enhancing the activity of antithrombin III (AT-III) and inhibiting the activity of coagulation factors IIa, IXa, and Xa, thereby preventing thrombus formation. However, the dosage of heparin is difficult to control precisely, and overdose can lead to serious bleeding complications such as gastrointestinal bleeding and intracranial hemorrhage, posing a direct threat to the patient's life.

[0005] Furthermore, long-term use of heparin may induce thrombocytopenia (HIT), further increasing the risk of bleeding and potentially leading to adverse consequences such as thrombosis. While low molecular weight heparin reduces the risk of bleeding to some extent, it cannot completely eliminate it, and its anticoagulant effect may be unsatisfactory in some patients with hypercoagulable states or anticoagulant resistance.

[0006] During hemodialysis, the contact between blood and extracorporeal circulation devices such as dialysis tubing and dialyzers, as well as changes in hemodynamics, can cause varying degrees of damage to the vascular endothelium. Vascular endothelial cells have important physiological functions, such as regulating vascular tone, maintaining blood rheological stability, inhibiting platelet aggregation and thrombus formation, and participating in the regulation of inflammatory responses. Once endothelial cells are damaged, their normal functions will be affected, leading to a series of adverse consequences.

[0007] Damaged vascular endothelial cells release various vasoactive substances and inflammatory factors, leading to vasoconstriction, increased platelet aggregation, and promotion of thrombus formation. This further aggravates renal ischemia and hypoxia, hindering the recovery of kidney function. Simultaneously, endothelial damage may also trigger the activation of inflammatory responses, causing inflammatory cell infiltration and the release of more inflammatory mediators, creating a vicious cycle and worsening the patient's condition.

[0008] Electrolyte balance is crucial for maintaining normal physiological functions. During hemodialysis, the electrolyte composition of the replacement fluid directly affects the patient's electrolyte levels. However, existing replacement fluid formulations have limitations in mimicking physiological electrolyte concentrations in the human body.

[0009] For example, the concentrations of electrolytes such as sodium, potassium, calcium, and magnesium in some replacement fluids may not perfectly match the patient's actual needs, leading to electrolyte imbalances during dialysis. Hyponatremia can cause symptoms such as dizziness, weakness, nausea, and vomiting, and in severe cases, it can lead to cerebral edema, seizures, or even coma. Hypernatremia, on the other hand, can cause thirst, polyuria, restlessness, and altered consciousness, increasing the cardiovascular burden on the patient and even inducing serious complications such as heart failure. Abnormal potassium ion concentrations can cause arrhythmias, endangering the patient's life. In addition, imbalances in ions such as calcium and magnesium can also adversely affect neuromuscular function and the cardiovascular system, impacting the patient's treatment outcomes and quality of life.

[0010] Hemodialysis patients commonly experience chronic inflammation due to impaired kidney function, toxin accumulation, and contact between blood and external substances during dialysis. Inflammation plays a crucial role in the development of hemodialysis-related complications; the risk of complications such as cardiovascular disease, malnutrition, and infections is closely related to the severity of the inflammatory response.

[0011] Current replacement fluids lack effective anti-inflammatory components and cannot specifically inhibit the release of inflammatory factors and the activation of inflammatory responses during dialysis. Persistent inflammation can further damage vascular endothelial function, promote the development of atherosclerosis, and increase the risk of cardiovascular events. Simultaneously, inflammation can affect patients' appetite and nutrient metabolism, leading to malnutrition, reducing immunity, and making patients more susceptible to complications such as infections, severely impacting their prognosis.

[0012] In summary, existing hemodialysis replacement fluids have many shortcomings in terms of anticoagulation, vascular protection, electrolyte balance, and inflammation control. These problems seriously affect the efficacy of hemodialysis treatment and patient prognosis. With the continuous advancement of medical technology and patients' increasing demands for treatment quality, it is particularly necessary to develop a new type of hemofiltration replacement fluid with superior performance.

[0013] Novel blood filtration replacement solutions should be able to effectively prevent coagulation while minimizing the risk of bleeding, protecting the vascular endothelium from damage, precisely maintaining electrolyte balance in patients, and effectively reducing inflammatory responses. This will help improve the efficacy of hemodialysis, reduce the occurrence of complications, improve patients' quality of life, prolong their survival, and provide safer, more effective, and personalized treatment options for patients with end-stage renal disease. Summary of the Invention

[0014] This invention aims to provide a novel blood filtration replacement fluid that overcomes the shortcomings of existing replacement fluids by carefully compounding anticoagulant components, vascular endothelial protectants, electrolytes, and anti-inflammatory factors. This ensures effective anticoagulation while reducing the risk of bleeding, protecting the vascular endothelium, maintaining electrolyte balance, and alleviating inflammatory responses, ultimately improving the treatment effect and prognosis of hemodialysis patients.

[0015] Technical solution

[0016] Formulation Composition: The blood filtration replacement solution of this invention contains 2000 IU of low molecular weight heparin calcium, 1.5 mg of prostaglandin E1, 5 mg of prostacyclin, 3 mmol of sodium citrate, 137 mmol of sodium, 2.0 mmol of potassium, 1.25 mmol of calcium, 0.5 mmol of magnesium, 112 mmol of chloride, 32 mmol of bicarbonate, 6.1 mmol of glucose, and 2.5 mg of interleukin-1 receptor antagonist per 1000 mL. The remainder is water for injection. All components are reliably sourced. Low molecular weight heparin calcium was purchased from Sanofi (Synthetyl), prostaglandin E1 from Sigma-Aldrich, and prostacyclin from GlaxoSmithKline (GSK) eprostol sodium, all meeting the corresponding quality standards.

[0017] Preparation method

[0018] Raw material preparation: Weigh each raw material accurately to ensure that the amount used is correct.

[0019] Dissolution and volume adjustment: Place the raw materials in a suitable container, add an appropriate amount of water for injection, stir at a specific stirring speed (600 rpm) for a sufficient time (45 minutes) to fully dissolve all components, and then adjust the volume to 1000 mL.

[0020] Pyrogen removal and aseptic packaging: Pyrogen removal is achieved by positive pressure filtration using a specific sterile microporous membrane (MilliporeSLGP033RB) with controlled pressure range (0.2-0.3MPa). The solution is then transferred to a medical polyvinyl chloride infusion bag (a product of Shandong Weigao Group) that has been sterilized with ethylene oxide and aseptically packaged in a Class 100 clean environment.

[0021] Beneficial effects

[0022] Balancing anticoagulation and bleeding risk: Low molecular weight heparin calcium and prostaglandin E1 work synergistically to effectively inhibit the coagulation process and reduce the risk of bleeding compared to traditional anticoagulants. The addition of prostacyclin further enhances the anticoagulant effect while helping to maintain normal vascular endothelial function and reduce vascular damage caused by anticoagulation.

[0023] Vascular protection: Prostaglandin E1 and prostacyclin effectively protect the vascular endothelium through mechanisms such as inhibiting platelet aggregation and relaxing vascular smooth muscle, reducing a series of complications caused by endothelial damage, such as thrombosis and vascular stenosis, helping to maintain the normal structure and function of blood vessels and improve the long-term effectiveness of hemodialysis.

[0024] Precise electrolyte balance: The carefully adjusted concentrations of sodium, potassium, calcium, magnesium, chloride, and bicarbonate ions closely match the electrolyte levels in the human body under physiological conditions. This effectively maintains the electrolyte balance in patients, preventing adverse reactions such as arrhythmia, muscle spasms, and acid-base imbalances caused by electrolyte disturbances during dialysis, and ensuring the stability of patients' physiological functions.

[0025] Sodium citrate reduces thrombin production by locally chelating calcium ions, thereby significantly enhancing the anticoagulant stability of the replacement fluid and reducing the risk of coagulation within the dialyzer and blood circuit. Furthermore, sodium citrate is metabolized into bicarbonate, improving the patient's local acid-base balance, particularly for patients with metabolic acidosis, further enhancing the safety and effectiveness of hemodialysis treatment.

[0026] Effectively reduces inflammatory response: Interleukin-1 receptor antagonists can specifically block the biological activity of the inflammatory factor interleukin-1, significantly reduce the damage to the body caused by inflammatory response during dialysis, reduce the risk of inflammation-related complications such as infection and cardiovascular disease, and benefit the patient's recovery and prognosis.

[0027] Comprehensive improvement of patient prognosis: Through the synergistic effects of the above-mentioned multiple aspects, the replacement fluid of the present invention can significantly improve the efficacy of hemodialysis, reduce the occurrence of complications, improve the quality of life of patients, and prolong the survival of patients, which has important clinical application value and broad application prospects. Attached Figure Description

[0028] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only for this invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0029] Figure 1 This is a schematic diagram of the high-performance liquid chromatography analysis of the components of this invention;

[0030] Figure 2 This is a schematic diagram of the drug toxicity test data of the present invention;

[0031] Figure 3 This is a schematic diagram of the preparation process of the present invention;

[0032] Figure 4 This is a schematic diagram of the clinical application system of the present invention. Detailed Implementation

[0033] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. It should also be noted that, to make the embodiments more comprehensive, the following embodiments are the best and preferred embodiments, and those skilled in the art can use other alternative methods to implement some well-known technologies; moreover, the accompanying drawings are only for more specific description of the embodiments and are not intended to specifically limit the present invention.

[0034] It should be noted that the use of terms such as "an embodiment," "an embodiment," "an exemplary embodiment," and "some embodiments" in the specification indicates that the described embodiment may include a specific feature, structure, or characteristic, but not every embodiment necessarily includes that specific feature, structure, or characteristic. Furthermore, when a specific feature, structure, or characteristic is described in connection with an embodiment, implementing such a feature, structure, or characteristic in conjunction with other embodiments (whether explicitly described or not) should be within the knowledge of those skilled in the art.

[0035] Generally, terms can be understood at least partly from their use in context. For example, depending at least partly on the context, the term "one or more" as used herein can be used to describe any feature, structure, or characteristic in a singular sense, or a combination of features, structures, or characteristics in a plural sense. Additionally, the term "based on" can be understood not necessarily to convey an exclusive set of factors, but rather, alternatively, depending at least partly on the context, to allow for the presence of other factors that are not necessarily explicitly described.

[0036] See Figures 1 to 4 As shown

[0037] The compounded anticoagulant, endothelial protectant, electrolytes, and anti-inflammatory factors were formulated into a replacement solution according to a specific ratio. The anticoagulant consisted of 2000 IU of low molecular weight heparin calcium and 11.5 mg of prostaglandin E; the endothelial protectant was 5 mg of prostacyclin; and sodium citrate was 3 mmol. The electrolytes included sodium 137 mmol / L, potassium 2.0 mmol / L, calcium 1.25 mmol / L, magnesium 0.5 mmol / L, chloride 112 mmol / L, bicarbonate 32 mmol / L, and glucose 6.1 mmol / L. The anti-inflammatory factor was an interleukin-1 receptor antagonist, 2.5 mg. All the above components were dissolved in water and brought to a final volume of 1000 mL. After pyrogen and heat removal treatment, the solution was aseptically packaged and sealed to obtain the final product.

[0038] How to use:

[0039] The prepared 1000 mL replacement fluid is mixed with the patient's blood at a replacement ratio of 1:1 to 2:1 for blood filtration. The filtration rate is adjusted according to the patient's condition, generally 100–200 mL / min.

[0040] Each hemodialysis session lasts 4 to 6 hours, 2 to 3 times a week. Coagulation function indicators and electrolyte levels are monitored regularly to assess the efficacy and adverse reactions.

[0041] For patients at high risk of complications such as severe infection or heart failure, the dosage of anticoagulants and anti-inflammatory factors may be appropriately increased, but close monitoring for complications such as bleeding is necessary.

[0042] Precautions:

[0043] This product is a prescription drug and should be used under the guidance of a physician. Do not use it on your own.

[0044] If severe bleeding, allergic reactions, or other adverse reactions occur, discontinue use immediately and seek medical attention promptly.

[0045] Patients with renal insufficiency and elderly patients should use this medication with caution, and the dosage should be adjusted if necessary.

[0046] The replacement fluid should be stored away from light and refrigerated at 2–8°C. It has a shelf life of 24 months.

[0047] Example 1

[0048] Raw material preparation:

[0049] Accurately weigh 2000 IU of low molecular weight heparin calcium (purchased from Sanofi, trade name: succinylhexidine, conforming to European Pharmacopoeia standards), 1.5 mg of prostaglandin E (purity ≥98%, purchased from Sigma-Aldrich), 5 mg of prostacyclin (manufacturer: GlaxoSmithKline, drug name: eprostacyclin sodium), 137 mmol of sodium (provided as analytical grade sodium chloride, purchased from Sinopharm Chemical Reagent Co., Ltd.), 2.0 mmol of potassium (analytical grade potassium chloride, Sinopharm Chemical Reagent Co., Ltd.), and 1.25 mmol of calcium (as anhydrous calcium chloride). Purchased from Tianjin Bodi Chemical Co., Ltd.: Magnesium 0.5 mmol (analytical grade magnesium chloride, Sinopharm Chemical Reagent Co., Ltd.), Chlorine 112 mmol (calculated based on the chlorine content of other salts, weighed appropriate chlorine-containing compound sodium chloride, Sinopharm Chemical Reagent Co., Ltd.), Bicarbonate 32 mmol (analytical grade sodium bicarbonate, Sinopharm Chemical Reagent Co., Ltd.), Glucose 6.1 mmol (analytical grade glucose powder, Sinopharm Chemical Reagent Co., Ltd.), Interleukin-1 receptor antagonist 2.5 mg (manufacturer: Sinopharm Biotechnology Co., Ltd., specification: 0.5 mg / vial).

[0050] Dissolution and Volume Adjustment: Place the above raw materials in a 1000mL glass container and add approximately 800mL of water for injection (prepared in-house using a two-stage reverse osmosis water purification process and a multi-effect distillation apparatus, conforming to the standards for water for injection in the Chinese Pharmacopoeia). Use a magnetic stirrer to stir at 600 rpm for 45 minutes to ensure complete dissolution of all components. Then, bring the volume to the 1000mL mark with water for injection to obtain the initial product of the displacement solution.

[0051] Pyrogen removal, pyrogen removal, and aseptic packaging: The initial replacement solution is filtered through a 0.22μm sterile microporous membrane (brand: Millipore, model: SLGP033RB) to remove pyrogens, using positive pressure filtration with the pressure controlled at 0.2-0.3MPa. The filtered solution is then transferred to medical PVC infusion bags (manufacturer: Shandong Weigao Group Medical Polymer Products Co., Ltd.) sterilized with ethylene oxide, and aseptically packaged in a Class 100 clean environment to obtain the final product. Sampling and testing of the finished product are conducted according to the relevant standards of the Chinese Pharmacopoeia to test for sterility and pyrogen removal. High-performance liquid chromatography (HPLC, instrument model: Agilent 1260 Infinity) is used to analyze the content of each component to ensure compliance with requirements.

[0052] Example 2

[0053] The dosages of low molecular weight heparin calcium were adjusted to 2500 IU, prostaglandin E1 to 1.2 mg, sodium citrate to 3 mmol, and prostacyclin to 6 mg. The sodium content was adjusted to 135 mmol / L (adjusted to the corresponding sodium salt concentration, Sinopharm Chemical Reagent Co., Ltd.), potassium to 2.2 mmol / L, calcium to 1.15 mmol / L, magnesium to 0.6 mmol / L, chloride to 110 mmol / L, bicarbonate to 30 mmol / L, glucose to 5.8 mmol / L, and the interleukin-1 receptor antagonist to 3 mg. The product was prepared according to the method described in Example 1, and sampling tests were performed to ensure product quality.

[0054] Clinical application examples

[0055] Example 1

[0056] Patient Selection and Treatment Preparation: A 45-year-old patient with chronic renal failure requiring hemodialysis, weighing 65 kg, with no history of severe bleeding disorders and normal cardiac function, was selected. Prior to hemofiltration, the patient underwent a comprehensive physical examination, and a complete blood count showed a red blood cell count of 4.5 × 10⁻⁶. 12 / L-5.5×10 12 / L, white blood cell count 4×10 9 / L-10×10 9 / L, platelet count 100×10 9 / L-300×10 9 The coagulation function indicators were PT 11-13 seconds, APTT 25-35 seconds, TT 14-21 seconds, and electrolyte levels were 3.5 mmol / L-5.5 mmol / L, sodium 135 mmol / L-145 mmol / L, chloride 96 mmol / L-106 mmol / L, calcium 2.1 mmol / L-2.55 mmol / L, and magnesium 0.7 mmol / L-1.1 mmol / L. These data were used as the baseline data before treatment.

[0057] Hemofiltration procedure: Connect 1000 mL of prepared replacement fluid to the Fresenius 4008S hemodialysis machine (manufactured by Fresenius Medical Products GmbH, Germany) at a replacement ratio of 1.5:1 for hemofiltration. The initial filtration rate is set to 150 mL / min. During treatment, closely monitor the patient's vital signs, maintaining blood pressure between 120 mmHg and 140 mmHg / 70 mmHg and 90 mmHg / 70 mmHg, heart rate between 60 and 100 beats per minute, and respiratory rate between 16 and 20 breaths per minute. Record the patient's vital signs every 30 minutes to ensure good patient tolerance.

[0058] Monitoring and Adjustment During Treatment: Blood samples were collected from the patient at 1, 2, 3, and 4 hours after treatment began to test coagulation function indicators (PT, APTT, TT), electrolyte levels (potassium, sodium, chloride, calcium, magnesium, bicarbonate), and inflammatory factor levels (C-reactive protein, pre-treatment level 5 mg / L-10 mg / L). Based on the test results, if abnormal changes in coagulation function indicators were found, such as PT or APTT prolonging more than 1.5 times the normal range, the proportion of anticoagulant components in the replacement fluid or the filtration rate could be adjusted appropriately. If electrolyte levels deviated, such as potassium ion concentration deviating from the normal range ±0.5 mmol / L, the replacement fluid formula should be adjusted promptly or appropriate supplemental treatment should be given.

[0059] Treatment Efficacy Evaluation and Summary: This hemodialysis treatment lasted for 4 hours. After the treatment, the patient underwent a comprehensive examination again, comparing coagulation function indicators, electrolyte levels, and clinical symptoms before and after treatment. The results showed that the patient's coagulation function indicators were stable, with PT at 12-13 seconds, APTT at 26-32 seconds, and TT at 15-19 seconds. Electrolyte levels remained within the normal range: potassium at 3.8 mmol / L-4.2 mmol / L, sodium at 138 mmol / L-142 mmol / L, chloride at 100 mmol / L-104 mmol / L, calcium at 2.2 mmol / L-2.4 mmol / L, and magnesium at 0.8 mmol / L-1.0 mmol / L. Clinical symptoms improved, indicating that this hemofiltration treatment was effective, and the use of replacement fluid did not cause significant adverse reactions.

[0060] Example 2 (Treatment for patients with comorbidities)

[0061] An elderly patient, aged 70 years and weighing 55 kg, with diabetic nephropathy, mild heart failure, and recent lung infection, was selected. A comprehensive examination was conducted before treatment to assess the patient's physical condition and risk factors.

[0062] Considering the patient's potential complications, the anticoagulant components in the replacement fluid were adjusted to 2800 IU of low molecular weight heparin calcium, 11.8 mg of prostaglandin E, and the anti-inflammatory factor interleukin-1 receptor antagonist was increased to 3.5 mg. Hemofiltration was performed using the replacement fluid at a 2:1 ratio with the patient's blood, with an initial filtration rate set at 120 mL / min.

[0063] During treatment, close monitoring of the patient is crucial. In addition to routine coagulation function and electrolyte level tests, close attention is paid to cardiac function indicators (BNP levels were 100 pg / mL-200 pg / mL before treatment) and blood glucose levels (fasting blood glucose 7 mmol / L-10 mmol / L). Vital signs are recorded every 20 minutes, and blood glucose is measured hourly. Treatment parameters are adjusted promptly based on the test results. If significant fluctuations in blood glucose are observed, exceeding 15 mmol / L or falling below 3.9 mmol / L, the glucose content in the replacement fluid is adjusted, or insulin therapy is administered. If cardiac function deteriorates, the infusion rate of the replacement fluid is appropriately reduced, or the anticoagulant dosage is adjusted.

[0064] After 5 hours of hemodialysis, the patient's various indicators improved to some extent. Heart failure symptoms were alleviated, BNP decreased to 50-100 pg / mL, the lung infection did not worsen, and coagulation function remained stable (PT 12-13.5 seconds, APTT 27-33 seconds, TT 16-20 seconds). Electrolyte balance was well maintained (potassium 3.7-4.3 mmol / L, sodium 136-140 mmol / L, chloride 101-105 mmol / L, calcium 2.15-2.35 mmol / L, magnesium 0.75-0.95 mmol / L). However, a small amount of nasal bleeding was observed during treatment. Anticoagulant use was immediately discontinued, and local hemostasis was achieved, stopping the bleeding. The patient's condition will continue to be closely monitored, and the subsequent treatment plan will be adjusted, such as appropriately reducing the anticoagulant dosage or changing the anticoagulation method.

[0065] Stability test examples

[0066] Accelerated testing: Three batches of prepared replacement fluid samples were placed in a constant temperature and humidity chamber (brand: Binder, model: MKF720) at 40℃±2℃ and 75%±5% relative humidity for 3 months. Samples were taken and tested at the end of the 0th, 1st, 2nd, and 3rd months. Tests included appearance (observation for discoloration, turbidity, precipitation, etc.), pH value (using a precision pH meter, model: Mettler-Toledo FE28), content of each component (anticoagulant, electrolytes, anti-inflammatory factors, analyzed using appropriate methods such as high performance liquid chromatography and ion chromatography; the content of each component should be 90%-110% of the labeled amount), sterility test (performed according to the sterility test method in the Chinese Pharmacopoeia; no bacterial growth should be observed), and pyrogen test (using the Limulus Amebocyte Lysate (LAL) reagent method; LAL reagent brand: Zhanjiang Andus, model: 0.125 EU / mL; the amount of endotoxin in each mL should be less than 0.5 EU).

[0067] Long-term test: Three batches of replacement fluid samples were placed in an environment with a temperature of 25°C ± 2°C and a relative humidity of 60% ± 10% (drug stability test chamber, brand: Yiheng, model: LHS-100CL) for 12 months. Samples were taken for testing at 0, 3, 6, 9, and 12 months respectively, and the test items were the same as those in the accelerated test.

[0068] Analysis of test results: The results of the accelerated test showed that there were no obvious changes in the appearance, pH value, and content of each component of the three batches of samples within 3 months, and the sterility test and pyrogen test were both qualified, indicating that the product has good stability under short-term high temperature and high humidity conditions. The results of the long-term test showed that all the indicators of the samples were stable within 12 months and met the quality standard requirements, indicating that the product can be stored stably under normal temperature and humidity conditions for at least 12 months, and the validity period can be tentatively set as 12 months (which can be further determined according to the stability data of a longer time in the future).

[0069] Examples of safety tests

[0070] Animal acute toxicity test

[0071] Selection and grouping of test animals: 60 healthy adult ICR mice, half male and half female, with a body weight of 20g - 25g were selected. They were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., and the animal license number is: SCXK (Beijing) 2021-0006. They were randomly divided into 6 groups, with 10 mice in each group.

[0072] Setting of dosing doses and dosing routes: Dosing groups were set with different multiples of the clinical equivalent dose, which were 5 times, 10 times, 15 times, 20 times, 25 times, and 30 times of the clinical equivalent dose respectively. The dosing route was tail vein injection, and the injection speed was controlled at 0.1 mL / 10g body weight - 0.2 mL / 10g body weight.

[0073] Observation indicators and test cycle: After dosing, the toxic reactions of the mice were closely observed, including behavioral activities (excitement, inhibition, convulsion), food and water intake, body weight changes, death conditions, etc. The observation cycle was 14 days, and the daily observation results were recorded. After 14 days, the surviving mice were dissected to observe the pathological changes of the main organs such as the heart, liver, spleen, lungs, and kidneys (using the hematoxylin-eosin staining method for tissue sections).

[0074] Result analysis: The test results showed that within the range of 5 to 15 times the clinical equivalent dose, no obvious toxic reactions occurred in the mice, their body weights increased normally, and their food and water intakes were normal. When the dose reached 20 times and above, some mice showed transient excitement and increased activity, and then returned to normal; 2 mice died in the highest dose group (30 times the clinical equivalent dose). The autopsy results showed that there was slight congestion in the main organs of the dead mice, but no obvious organic lesions were found. The median lethal dose (LD50) of this replacement fluid for mice and its 95% confidence limit were calculated to provide a reference basis for clinical safe medication (specific data are shown in Table 1).

[0075] Table 1:

[0076]

[0077] Long-term toxicity test of animals

[0078] Test animals and grouping: 30 healthy adult SD rats, 15 males and 15 females, with body weights of 200 g - 250 g, were selected. They were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., and the animal license number was: SCXK (Beijing) 2021 - 0006. They were randomly divided into 3 groups, with 10 rats in each group, namely the low-dose group (1 times the clinical equivalent dose), the medium-dose group (3 times the clinical equivalent dose), and the high-dose group (5 times the clinical equivalent dose).

[0079] Administration method and cycle: The drug was administered by tail vein injection, 3 times a week for 12 consecutive weeks.

[0080] Observation indicators: During the drug administration period, the body weights and body temperatures of the animals were measured weekly, and the appearance signs, behavioral activities, food and water intakes of the animals were observed. Blood samples were collected every 4 weeks to detect blood routine (including white blood cells, red blood cells, platelets, etc., using a fully automatic blood analyzer, brand: Sysmex, model: XN - 1000), coagulation function indicators (PT, APTT, TT, coagulation analyzer brand: Stago, model: STA - RMax), liver and kidney function indicators (ALT, AST, BUN, Cr, etc., biochemical analyzer brand: Hitachi, model: 7180), electrolyte levels (potassium, sodium, chlorine, calcium, magnesium, etc., electrolyte analyzer brand: Roche, model: Cobas6000e), etc.; electrocardiogram examinations were performed every 8 weeks (electrocardiograph brand: Nihon Kohden, model: ECG - 1350P); after 12 weeks of drug administration, all animals were dissected comprehensively to observe the gross morphology and histological changes of each organ.

[0081] Results Assessment: The experimental results showed that the low-dose group animals maintained basically normal indicators throughout the experiment, with no obvious toxic reactions. In the medium-dose group, some animals showed mild elevations in liver function indicators in the later stages of administration, with ALT elevation not exceeding 1.5 times the upper limit of normal and AST elevation not exceeding 1.3 times the upper limit of normal, but these gradually returned to normal after drug withdrawal. In the high-dose group, animals experienced slow weight gain and mild anemia during administration, with red blood cell count decreasing by no more than 10% of the lower limit of normal and hemoglobin content decreasing by no more than 15% of the lower limit of normal. Some animals showed T-wave changes on electrocardiograms, but no significant arrhythmias were found. Autopsy results showed that some animals in the high-dose group had mild fatty degeneration in the liver, with the proportion of fatty hepatocytes not exceeding 5%, and mild renal tubular dilation in the kidneys, with the proportion of renal tubular dilation not exceeding 10%, but all were reversible changes. Overall assessment suggests that within the clinically equivalent dose range, long-term use of this replacement fluid has good safety; however, high doses may have certain potential effects on the liver, kidneys, and hematologic system, requiring close monitoring of relevant indicators during clinical application.

[0082] The specific parameters and dosages used in the technical solution are preferred embodiments of the present invention, but should not be construed as limitations on the invention. Those skilled in the art can make appropriate adjustments within the scope of the invention according to actual needs, and all such adjustments should be considered within the protection scope of the invention. The significant effect of the present invention is that it provides a hemofiltration replacement fluid that combines multiple anticoagulant components, balances anticoagulation and vascular protection, simulates human electrolyte balance, and reduces inflammatory responses. Compared with existing technologies, it has superior anticoagulant effects, lower bleeding risk, better vascular protection, and electrolyte balance capabilities, significantly improving the prognosis of hemodialysis patients, and has significant clinical application value and promising prospects for wider application.

[0083] This invention encompasses any substitutions, modifications, equivalent methods, and solutions made within the spirit and scope of this invention. To provide the public with a thorough understanding of this invention, specific details are described in detail in the following preferred embodiments; however, those skilled in the art will fully understand the invention even without these details. Furthermore, to avoid unnecessary misunderstanding of the essence of this invention, well-known methods, processes, procedures, components, and circuits are not described in detail.

[0084] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. An anticoagulation-specific hemofiltration replacement fluid, characterized in that, The composition comprises the following ingredients per 1000 mL of replacement fluid: Low molecular weight heparin calcium: 2000 IU; Prostaglandin E1: 1.5 mg; Prostacyclin: 5 mg; Sodium citrate: 3 mmol; Sodium ions: 134 mmol (including sodium ions in sodium citrate); Potassium ions: 2.0 mmol; Calcium ions: 1.25 mmol; Magnesium ions: 0.5 mmol; Chloride ions: 112 mmol; Bicarbonate: 32 mmol; Glucose: 6.1 mmol; Interleukin-1 receptor antagonist: 2.5 mg; The rest is water for injection, the replacement fluid is treated by sterile filtration, and is packaged in a sterile medical container.

2. The anticoagulation special blood filtration replacement fluid according to claim 1, characterized in that, The replacement fluid is suitable for hemodialysis of end-stage renal disease patients, can effectively anticoagulate and reduce the risk of bleeding.

3. The anticoagulation special blood filtration replacement fluid according to claim 1, characterized in that, The preparation method of the replacement fluid comprises the following steps: Accurately weigh each ingredient, add an appropriate amount of water for injection, and stir to dissolve; Use a 0.22 μm micropore filter to filter out pyrogens; Sterile filling and sealed packaging are carried out in a hundred-level clean environment.

4. The method of claim 1, wherein the anti-coagulation dedicated hemofiltration replacement fluid is prepared by adding 0.9 to 1.1 g of sodium citrate to 1,000 ml of a 5% dextrose solution. The method comprises the following steps: Accurately weigh each ingredient raw material according to the proportion, add 800 mL of water for injection, use a magnetic stirrer to stir at a speed of 600 revolutions per minute for 45 minutes, and use water for injection to make up to 1000 mL; Positive pressure filtration is carried out through a sterilization micropore filter with a pore size of 0.22 microns, and the filtration pressure is controlled within the range of 0.2 to 0.3 megapascals; The filtered solution is transferred to a medical polyvinyl chloride infusion bag treated by ethylene oxide sterilization, and is sealed and packaged in a hundred-level clean environment.

5. The anticoagulation-specific hemofiltration substitution solution according to claim 1, characterized in that, The administration parameters in clinical application are: The replacement ratio of replacement fluid to patient's blood is 1:1 to 2:1; The filtration rate is set to 100 to 200 mL per minute; The single treatment time is 4 to 6 hours; The administration frequency is 2 to 3 times per week.

6. The quality standard of the anticoagulant special blood filtration replacement fluid according to claim 1, wherein: The content of each active ingredient ranges from 90% to 110% of the labeled amount; The bacterial test result is sterile growth; The endotoxin content is less than 0.5 endotoxin units per milliliter; The pH value ranges from 6.8 to 7.

4.

7. The anticoagulant special blood filtration replacement fluid according to claim 1, wherein: The storage temperature is 2 to 8 degrees Celsius; The packaging container is a medical polyvinyl chloride infusion bag sterilized by ethylene oxide; The packaging specification is 1000 mL per bag; The labeled validity period is 24 months.

8. The toxicity test scheme of the anticoagulant special blood filtration replacement fluid according to claim 1, wherein: The acute toxicity test sets the administration dose to be 5 times, 10 times, 15 times, 20 times, 25 times, and 30 times of the clinical equivalent dose, and the observation period is 14 days; The long-term toxicity test sets the administration dose to be 1 time, 3 times, and 5 times of the clinical equivalent dose, the administration period is 12 weeks, and the administration frequency is 3 times per week; The detection indexes include blood routine, coagulation function indexes PT, APTT, TT, liver function indexes ALT, AST, kidney function indexes BUN, Cr, and electrolytes potassium, sodium, chlorine, calcium, and magnesium.

9. The stability test condition of the anticoagulant special blood filtration replacement fluid according to claim 1, wherein: The accelerated test conditions are temperature 40 ± 2 degrees Celsius, relative humidity 75% ± 5%, the test period is 3 months, and the sampling time points are the end of the 0th, 1st, 2nd and 3rd months; The long-term test conditions are temperature 25 ± 2 degrees Celsius, relative humidity 60% ± 10%, the test period is 12 months, and the sampling time points are the 0th, 3rd, 6th, 9th and 12th months; The detection items include appearance change, pH value determination, effective ingredient content determination, sterility test and pyrogen test.

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