A dialysis solution containing human albumin and glucose and preparation method thereof

Abstract

The present invention relates to the field of biomedicine, and in particular to a dialysate containing human albumin and glucose and a preparation method thereof. The dialysate of the present invention comprises human albumin, glucose, an osmotic pressure regulator, a buffer, and a stabilizer. Among them, the human albumin content is 4.4% to 12.5% ​​(w / w), and the glucose content is 1% to 2.5%. The present invention provides a low-cost, high-quality dialysate prescription and preparation process, which has good biocompatibility, can avoid the risk of side effects caused by patients during treatment, and plays a key role in the removal of metabolites such as bilirubin and creatinine in patients with renal failure.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine, and in particular to a dialysate containing human albumin and glucose and a preparation method thereof. Background Art

[0002] The most commonly used treatment for renal failure and uremia is dialysis, which is the removal of waste toxins and excess water from the body. Patients can maintain their lives using dialysis technology, and in developed countries, the dialysis age can reach more than 20 years. There are two types of dialysis technology: one is hemodialysis and the other is peritoneal dialysis. Peritoneal dialysis is a new technology that our country is promoting and is suitable for my country's national conditions. This technology is only in the development stage in my country, but compared with hemodialysis, it has the characteristics of being able to be treated at home, so it can greatly improve the quality of life of patients, and patients can go to work, work and study like normal people. Since there is no need to frequently go back and forth to the hospital, it is also suitable for kidney patients in remote rural areas, so my country is sparing no effort to promote this treatment technology.

[0003] The principle of peritoneal dialysis technology is to exchange metabolites in the body into peritoneal dialysis fluid through the peritoneum, and then discharge the dialysis fluid out of the body. Peritoneal dialysis fluid is a high osmotic pressure solution that works through the principle of osmotic pressure.

[0004] The peritoneal dialysis fluid currently on the market in my country is a dialysis fluid with sugar as the main raw material. The use of monosaccharides or polysaccharides as the raw materials of dialysis fluid has poor biocompatibility and a relatively complicated production process. At the same time, the existing peritoneal dialysis fluid with sugar as the main raw material is often divided into liquid A and liquid B, which need to be stored separately and mixed in proportion when used, and the operation process is complicated.

[0005] Especially for the current dialysate which is mainly composed of glucose, the higher the glucose concentration, the better the dialysis effect. However, the sugar-containing peritoneal dialysis fluid will increase the sugar load of the patient's body and more or less interfere with the blood sugar level. High-sugar dialysate will also increase the chance of peritoneal damage.

[0006] With the rapid development of medical technology, the important position of human albumin in clinical medicine is increasingly recognized by the biomedical field, because it can scavenge free radicals and increase the solubility of long-chain fatty acids; it can also be used as a delivery system for anti-tumor drugs. This provides a new research direction for the development of a new type of dialysate with good compatibility, simple preparation and convenient use. Summary of the invention

[0007] To solve the above problems, the present invention provides a dialysate containing human albumin and glucose and a preparation method thereof, which improves the biocompatibility and effectiveness of the dialysate, reduces the sugar load of high sugar on the body, and reduces damage to the peritoneum, which is of great significance for toxin removal and health improvement.

[0008] On the one hand, the present invention provides a dialysate containing human albumin, comprising: human albumin, an antioxidant, an osmotic pressure regulator, a buffer, a stabilizer and an antibacterial agent, wherein the content of human albumin is 4.4% to 12.5% ​​(w / w); the content of glucose is 1% to 2.5% (w / w); the content of the osmotic pressure regulator is 0.2% to 0.8% (w / w); the content of the buffer is 0.01% to 0.1% (w / w); and the content of the stabilizer is 0.2% to 1% (w / w).

[0009] In an embodiment of the present invention, the human albumin includes one or more of blood-derived human albumin and recombinant human albumin. Preferably, the human albumin is recombinant human albumin. Among them, blood-derived human albumin refers to albumin separated and purified from healthy human blood, plasma or specific immune plasma; recombinant human albumin refers to albumin prepared by recombinant DNA technology for the treatment and prevention of diseases.

[0010] In an embodiment of the present invention, the osmotic pressure regulator includes sodium chloride, and the content of the osmotic pressure regulator is

[0011] 0.2% to 0.8% (w / w). Preferably, the osmotic pressure regulator may also include a trace amount of calcium chloride or magnesium chloride, wherein the content of calcium chloride is 0.01 to 0.03% (w / w), and the content of magnesium chloride is 0.002 to 0.006% (w / w).

[0012] In an embodiment of the present invention, the buffer comprises one or more of acetate and lactate, bicarbonate, pyruvate, and citrate, and the content of the buffer is 0.01% to 0.1% (w / w). Preferably, the buffer is lactate, and more preferably, the buffer is sodium lactate.

[0013] In an embodiment of the present invention, the stabilizer includes one or more of polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, and poloxamer 407, and the content of the stabilizer is 0.2% to 1% (w / w). Preferably, the stabilizer is a polysorbate series, and more preferably, the stabilizer is polysorbate 80.

[0014] In an embodiment of the present invention, the dialysate further comprises an antioxidant, wherein the content of the antioxidant is 0.01-0.1% (w / w). The antioxidant comprises one or more of sodium sulfite and sodium bisulfite. Preferably, the antioxidant is sodium sulfite.

[0015] In a preferred embodiment of the present invention, the permeate comprises: 4.4% to 12.5% ​​(w / w) human albumin; 1% to 2.5% (w / w) glucose; 0.01% to 0.1% (w / w) sodium lactate; 0.2% to 0.8% (w / w) sodium chloride; 0.2% to 1% (w / w) polysorbate 80; 0.01% to 0.1% (w / w) sodium sulfite; and the remainder is water for injection.

[0016] In an embodiment of the present invention, the pH value of the dialysate of the present invention is 6.4-7.4, and the osmotic pressure is 300-380 mOsm / Kg.

[0017] A second aspect of the present invention provides a method for preparing the dialysate, comprising the following steps:

[0018] Sterilization: The salt components of each component are sterilized at high temperature, the temperature is 115℃~121℃, and the sterilization time is 15~60min; in the embodiment of the present invention, the salt components include sulfite, bisulfite, chloride, acetate and lactate, bicarbonate, pyruvate, citrate or more, preferably, the salt components include sodium lactate, sodium chloride, sodium sulfite. In the preferred embodiment of the present invention, the sterilization temperature is 121℃, and the sterilization time is 30min.

[0019] Preparation: Add each component according to the content ratio into the sterilized water for injection, dissolve and stir evenly, and sterilize and filter with a filter of a specific pore size; in the embodiment of the present invention, add each component per 1000ml, and the mass of each component is: human albumin: 44-125g, glucose: 10-25g, sodium lactate: 0.1-1.0mg, sodium chloride: 2-8mg, polysorbate 80: 2-10mg, sodium sulfite: 0.1-1.0mg, and the rest is water for injection. In the embodiment of the present invention, the pore size of the filter is filtered through 0.45μm and 0.22μm respectively.

[0020] Filling: Under a clean bench, fill the filtrate obtained in step 2) into a medical dialysis bag, put the filled dialysis bag into an outer bag with a high barrier, evacuate for 0.5 to 2 minutes and fill with nitrogen for 1 to 10 minutes, and package to obtain the dialysate. In an embodiment of the present invention, the medical dialysis bag in step (3) is made of non-PVC material.

[0021] The beneficial effects of the present invention include but are not limited to:

[0022] 1. The present invention provides a dialysate containing human albumin and glucose, which uses human albumin and glucose as main raw materials. Compared with the existing dialysate using only sugars as main raw materials, the present invention has simpler composition, better biocompatibility, increased ultrafiltration function, enhanced dialysis effect, and achieves therapeutic effect.

[0023] 2. The dialysate containing human albumin and glucose provided by the present invention has osmotic pressure provided by human albumin and glucose, which can significantly reduce the use of high-concentration glucose dialysate, reduce the interference with human blood sugar, and also reduce the damage of high-sugar dialysate to the peritoneum.

[0024] 3. The dialysate of the present invention is easy to store and does not need to be stored separately according to pH value or to prevent reactions between components. It does not need to be mixed according to a ratio when used, and is easy to operate.

[0025] 4. The present invention also provides a method for preparing a dialysate containing human albumin and glucose, which has a simple production process and has a high application prospect.

[0026] 5. The dialysis fluid containing human albumin and glucose provided by the present invention has lower degradation products and higher stability than the dialysis fluid based on glucose. DETAILED DESCRIPTION

[0027] The present invention is further described in detail below through specific implementation modes, but it is only used to help understand the present invention so that professionals in the field can implement or use the present invention, and does not constitute any limitation to the present invention.

[0028] Definition: Hematogenous human albumin refers to albumin separated and purified from healthy human blood, plasma or specific immune plasma;

[0029] Recombinant human albumin refers to albumin prepared using recombinant DNA technology for treatment and prevention.

[0030] Source of materials: Recombinant human albumin: produced by Tonghua Anrui Biopharmaceutical Co., Ltd., product number: ART101S;

[0031] Blood-derived human albumin: purchased from Guangdong Weilun Biopharmaceutical Co., Ltd.;

[0032] SD rats: purchased from Jiangsu Jicui Yaokang Biotechnology Co., Ltd.;

[0033] Glucose peritoneal dialysis fluid: purchased from Huaren Pharmaceutical Co., Ltd. and Shijiazhuang Siyao Co., Ltd.;

[0034] Unless otherwise specified, other reagents and instruments used were commercially available.

[0035] Example 1 Preparation of Dialysate

[0036] The composition of each 1000 ml of dialysate by weight is as shown in Table 1. The rest is sterile water for injection.

[0037] Table 1 Composition of various dialysates containing human albumin and glucose

[0038]

[0039] Table 1 continued

[0040]

[0041] The specific steps for preparing 1000 ml of dialysate are as follows:

[0042] 1) Use a sterilized grade A measuring cylinder to measure 1000 ml of sterilized water for injection;

[0043] 2) According to the composition of each formula, weigh the corresponding mass of various salt components for high temperature sterilization at 121°C for 30 minutes;

[0044] 3) According to the formula, weigh: human albumin dry powder, glucose, polysorbate 80 and other non-salt components, and the sterilized salt components obtained in 2) into a sterilized beaker, add 1000 ml of sterilized water for injection, dissolve, stir evenly, and filter through a 0.45 μm filter membrane to remove particles, and then use a 0.22 μm filter membrane for sterilization filtration to obtain a filtrate, fill the filtrate into a medical dialysis bag in an ultra-clean workbench, put the filled medical dialysis bag into an outer bag with a high barrier, evacuate for 0.5 to 2 minutes and fill with nitrogen for 1 to 10 minutes, and package to obtain a dialysate containing human albumin and glucose of the corresponding serial number.

[0045] Example 2 Peritoneal influence test

[0046] Experimental animals: SD rats, male, weighing 200±10g, a total of 260 rats, were randomly divided into 52 groups, 5 rats in each group, of which groups 1 to 45 were injected with dialysates containing human albumin of formulas No. 1 to 45 in Table 1. Groups 46 to 52 were the control groups, and the specific dialysate information is shown in Table 2.

[0047] Table 2 Dialysate information of the control group

[0048] Group Dialysate 46 Normal saline 47 Huaren Pharmaceutical Co., Ltd. 1.5% G peritoneal dialysis solution (lactate) 48 Huaren Pharmaceutical Co., Ltd. 2.5% G peritoneal dialysis solution (lactate) 49 Huaren Pharmaceutical Co., Ltd. 4.25% G peritoneal dialysis solution (lactate) 50 Shijiazhuang Siyao Co., Ltd. 1.5% G peritoneal dialysis solution (lactate) 51 Shijiazhuang Siyao Co., Ltd. 2.5% G peritoneal dialysis solution (lactate) 52 Shijiazhuang Siyao Co., Ltd. 4.25%G peritoneal dialysis solution (lactate)

[0049] Pharmacodynamics experiment of recombinant human albumin and blood-derived human albumin dialysate

[0050] The corresponding dialysate (100 ml / kg) was injected intraperitoneally into the SD rats of each group once a day for 6 consecutive weeks until the dialysis was terminated.

[0051] Determination of filtration capacity

[0052] Within 24 hours after the end of ultradialysis, each SD rat in groups 1 to 52 was anesthetized and injected with 0.9% saline (100 ml / kg) intraperitoneally. Four hours later, the abdominal wall was cut open along the linea alba, the peritoneal fluid was measured, and the volume of the total dialysis effluent (ml) was measured. The experimental results are shown in Table 3.

[0053] Ultrafiltration volume: Ultrafiltration volume = final water output - 25mL.

[0054] Determination of bilirubin, creatinine concentration and urea nitrogen content

[0055] Blood samples and peritoneal fluid were collected from each SD rat at 0h and 4h after dialysis. The blood samples were centrifuged at 4000rpm / min for 15min, and the peritoneal dialysis fluid samples were centrifuged at 2000rpm / min for 5min. The bilirubin, creatinine and urea nitrogen concentrations in the effluent and plasma were measured. The experimental results are shown in Table 3.

[0056] Bilirubin clearance = [total dialysate outflow volume ml x (ratio of bilirubin concentration in dialysate outflow to bilirubin concentration in plasma)];

[0057] Creatinine clearance = [total dialysate outflow volume ml x (ratio of creatinine concentration in dialysate outflow to creatinine concentration in plasma)];

[0058] Urea nitrogen clearance = [total dialysate outflow volume ml x (ratio of urea nitrogen concentration in dialysate outflow to urea nitrogen concentration in plasma)].

[0059] Table 3 Results of ultrafiltration test and bilirubin, creatinine concentration and urea nitrogen content of dialysate in each group

[0060]

[0061]

[0062] As can be seen from Table 3, the ultrafiltration volume of SD rats in the experimental results of the dialysate containing recombinant human albumin of the present invention and the experimental groups (Groups 1 to 45) containing the dialysate containing hematogenous human albumin are significantly higher than that of Group 46 dialyzed with 0.9% saline and the control group (Groups 47 to 52) using commercially available sugar-containing peritoneal dialysis fluid. The ultrafiltration volume results of the recombinant human albumin dialysate of the present invention are similar to those of the hematogenous human albumin dialysate, indicating that the recombinant human albumin dialysate has the same efficacy as the hematogenous human albumin dialysate and is closer to normal physiological conditions.

[0063] The clearance capabilities of SD rats in groups 1 to 45 that were dialyzed with the dialysate containing human albumin and glucose described in the present invention were higher than those of rats in group 46 that were dialyzed with 0.9% saline dialysate and rats in groups 47 to 52 that were dialyzed with carbohydrate dialysate; and the dialysate made with our company's recombinant human albumin performed better in removing toxins than the dialysate made with hematogenous human albumin.

[0064] Example 3 Determination of glucose degradation products

[0065] 5-Hydroxymethylfurfural:

[0066] Preparation of test solution: Accurately measure an appropriate amount of a dialysate containing human albumin and glucose prepared in Example 1 above and a commercially available glucose dialysate (equivalent to approximately 0.1 g of glucose), place them in a 10 ml volumetric flask, add purified water to dilute to the scale, and shake well to obtain the test solution.

[0067] Preparation of reference solution: Accurately measure 0.1 g of 5-hydroxymethylfurfural reference substance, place it in a 10 ml volumetric flask, add purified water to dilute to the scale, shake well, and obtain the reference solution.

[0068] Chromatographic conditions: Thermo Vanquish Core, Column: Waters Peptide BEH C18: 1.7 μm, 4.6 mm × 300 mm, mobile phase A is 0.1% formic acid-water, mobile phase B is 0.1% formic acid-acetonitrile, column temperature: 40°C, injection volume: 10 μl. Elution gradient is shown in Table 4.

[0069] Table 4 Elution gradient

[0070]

[0071]

[0072] Determination method: Accurately measure the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatogram. Calculate the peak area according to the external standard method. The experimental results are shown in Table 5.

[0073] Table 5 Determination of glucose degradation products Experimental results

[0074]

[0075]

[0076] It can be seen from the data in Table 5 that the content of 5-hydroxymethylfurfural in the dialysate containing human albumin and glucose (groups 1-45) of the present invention is lower than that in the commercially available glucose dialysate (groups 47-52), indicating that the dialysate containing human albumin and glucose prepared by the present invention has lower glucose degradation products and higher stability.

[0077] The production method of the invention has simple process, convenient operation and easy commercial production. The prepared product is a dialysate containing human albumin and glucose, which is suitable for patients with renal failure. Compared with the dialysate mainly composed of sugars, it has better clearance ability, good biocompatibility and efficacy, and has broad development prospects.

[0078] The above description is only an embodiment of the present invention. The protection scope of the present invention is not limited by these specific embodiments, but is determined by the claims of the present invention. For those skilled in the art, the present invention may have various changes and variations. Any modification, equivalent substitution, improvement, etc. made within the technical ideas and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A peritoneal dialysis solution containing human albumin and glucose, characterized in that: The peritoneal dialysis fluid comprises: human albumin, glucose, an osmotic pressure regulator, a buffer and a stabilizer, wherein: The content of human albumin is 4.4%~12.5% ​​(w / w); The content of glucose is 1%~2.5% (w / w); The content of the osmotic pressure regulator is 0.0002% to 0.0008% (w / w), and the osmotic pressure regulator includes sodium chloride; The content of the buffer is 0.00001% to 0.0001% (w / w), and the buffer is one of sodium acetate, sodium lactate, sodium bicarbonate, sodium pyruvate, and sodium citrate; The content of the stabilizer is 0.0002% to 0.001% (w / w), and the stabilizer is one of polysorbate 80, polysorbate 20, polysorbate 40, polysorbate 60, and poloxamer 407.

2. The peritoneal dialysis fluid according to claim 1, characterized in that The human albumin is hematogenous human albumin or recombinant human albumin.

3. The peritoneal dialysis fluid according to claim 1, characterized in that The osmotic pressure regulator also includes calcium chloride or magnesium chloride.

4. The peritoneal dialysis fluid according to claim 1, characterized in that The peritoneal dialysis fluid also includes an antioxidant, which is sodium sulfite or sodium bisulfite, and the mass fraction of the antioxidant is 0.00001% to 0.0001% (w / w).

5. The peritoneal dialysis fluid according to claim 1, characterized in that The peritoneal dialysis fluid includes: 4.4%~12.5% ​​(w / w) human albumin; 1%~2.5% (w / w) glucose; 0.00001%~0.0001% (w / w) sodium lactate; 0.0002%~0.0008% (w / w) sodium chloride; 0.0002%~0.001% (w / w) polysorbate 80; 0.00001%~0.0001% (w / w) sodium sulfite, and the balance is water for injection.

6. A method for preparing the peritoneal dialysis fluid according to any one of claims 1 to 5, characterized in that: The steps include: 1) Sterilization: sterilize the salt components at high temperature, temperature: 115℃~121℃, sterilization time: 15~60min; 2) Preparation: Add each component according to the content ratio into sterilized water for injection, dissolve and stir evenly, and sterilize and filter with filters with pore sizes of 0.45µm and 0.22µm; 3) Filling: Under a clean bench, fill the filtrate obtained in step 2) into a medical dialysis bag, put the filled dialysis bag into an outer bag with a high barrier, evacuate for 0.5 to 2 minutes, fill with nitrogen for 1 to 10 minutes, and seal to obtain the dialysate.

7. The method according to claim 6, characterized in that The medical dialysis bag described in step 3) is a dialysis bag made of non-PVC material.

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