Lung perfusion preservation solution for use at room temperature and method for its preparation

Abstract

The application discloses a lung perfusion preservation solution used at normal temperature and a preparation method thereof, and belongs to the technical field of organ preservation. The lung perfusion preservation solution used at normal temperature is composed of the following components: dextran 40, sodium chloride, potassium chloride, calcium chloride dihydrate, sodium phosphate monobasic monohydrate, sodium bicarbonate, magnesium chloride, anhydrous alpha-D-glucose, xylitol, human serum albumin, ligustrazine, furosemide, sodium hydroxide and sterile water. The lung perfusion preservation solution prepared by the application can preserve an isolated lung for a long time, effectively reduces the risk of lung edema, greatly reduces the cost and alleviates the economic burden of lung transplant patients through the interaction of human serum albumin, inorganic salts, ligustrazine, furosemide and xylitol.

Description

Technical Field

[0001] This invention relates to the field of organ preservation technology, and in particular to an ex vivo perfusion preservation solution for lungs that can be used at room temperature and its preparation method. Background Technology

[0002] Transplantation is the preferred treatment for organ failure, and deceased organ donation is the main source of organs for transplantation in my country. Donated organs may suffer from functional impairment and prolonged warm or cold ischemia, leading to damage and affecting transplant prognosis. Current technologies have found that perfusion preservation solutions can prolong the preservation time and postoperative recovery of ex vivo organs, making perfusion preservation solutions a hot research topic in organ preservation and transplantation.

[0003] The primary perfusion preservation solution for lung transplantation is Steen solution from Toronto. Steen solution, based on extracellular fluid characteristics, has a low ion concentration, low potassium, contains dextran, possesses physiological osmotic pressure, and maintains a high colloid osmotic pressure with a certain protein concentration. It can remove excess water from the lung interstitium and alveoli, protecting the integrity of the alveolar capillary membrane. However, this solution is not domestically produced, and imported products are expensive, increasing the financial burden on lung transplant patients. Furthermore, excised lungs using Steen solution can only be maintained for 12 hours, and there are potential risks of infection and edema. Summary of the Invention

[0004] To address the problems existing in the prior art, this invention provides a lung ex vivo perfusion preservation solution that can be used at room temperature and its preparation method. The lung ex vivo perfusion preservation solution of this invention, which can be used at room temperature, can significantly reduce the risk of pulmonary edema and infection, and reduce costs.

[0005] To solve the above-mentioned technical problems, the present invention provides the following technical solution:

[0006] On one hand, the present invention provides an ex vivo lung perfusion preservation solution that can be used at room temperature, comprising the following components:

[0007] Dextran 40, sodium chloride, potassium chloride, calcium chloride dihydrate, sodium dihydrogen phosphate monohydrate, sodium bicarbonate, magnesium chloride, anhydrous α-D-glucose, xylitol, human serum albumin, ligustrazine, furosemide, sodium hydroxide, and sterile water.

[0008] In the ex vivo lung perfusion preservation solution that can be used at room temperature, the concentration of dextran 40 is 4-6 g / L; the concentration of sodium chloride is 4-6 g / L; the concentration of potassium chloride is 0.4-0.6 g / L; the concentration of calcium chloride dihydrate is 0.2-0.25 g / L; the concentration of sodium dihydrogen phosphate monohydrate is 0.1-0.2 g / L; the concentration of sodium bicarbonate is 0.8-1.4 g / L; the concentration of magnesium chloride is 0.1-0.15 g / L; the concentration of anhydrous α-D-glucose is 1.5-2 g / L; the concentration of xylitol is 0.4-0.6 g / L; the concentration of human serum albumin is 66-72 g / L; the concentration of tetramethylpyrazine is 0.1-0.17 g / L; and the concentration of furosemide is 0.01-0.03 g / L.

[0009] Preferably, in the above-mentioned lung ex vivo perfusion preservation solution that can be used at room temperature, the concentration of dextran 40 is 5 g / L; the concentration of sodium chloride is 5 g / L; the concentration of potassium chloride is 0.54 g / L; the concentration of calcium chloride dihydrate is 0.22 g / L; the concentration of sodium dihydrogen phosphate monohydrate is 0.14 g / L; the concentration of sodium bicarbonate is 1.26 g / L; the concentration of magnesium chloride is 0.11 g / L; the concentration of anhydrous α-D-glucose is 1.8 g / L; the concentration of xylitol is 0.4 g / L; the concentration of human serum albumin is 70 g / L; the concentration of tetramethylpyrazine is 0.14 g / L; and the concentration of furosemide is 0.02 g / L.

[0010] Furthermore, the pH of the ex vivo lung perfusion preservation solution, which can be used at room temperature, is 7.2-7.4.

[0011] On the other hand, the present invention provides a method for preparing the above-mentioned lung ex vivo perfusion preservation solution that can be used at room temperature, comprising:

[0012] 1) Provide raw materials according to the above-mentioned components of the lung ex vivo perfusion preservation solution that can be used at room temperature;

[0013] 2) Heat and dissolve dextran 40 and ligustrazine in sterile water for later use;

[0014] 3) Dissolve sodium chloride, potassium chloride, sodium dihydrogen phosphate monohydrate, sodium bicarbonate, magnesium chloride, anhydrous α-D-glucose, and xylitol in sterile water; titrate with sodium hydroxide to pH 7.4, and set aside; the concentration of sodium hydroxide is 1 mol / L.

[0015] 4) Mix the solution prepared in step 2) with the solution prepared in step 3), then add calcium chloride dihydrate, human serum albumin, and furosemide, and adjust the volume with sterile water. Then adjust the pH to 7.2-7.4 with sodium hydroxide and set aside.

[0016] 5) Filter the above liquid using a sterile filter, dispense and seal for storage at 4°C.

[0017] Furthermore, the present invention also provides the application of the above-mentioned lung perfusion preservation solution, which can be used at room temperature, in the preservation of isolated lungs.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] This invention improves Steen solution. Human serum albumin provides colloid osmotic pressure, preventing pulmonary edema during perfusion. The addition of tetramethylpyrazine and furosemide further reduces pulmonary edema and improves the functional stability of the isolated lung. Furthermore, xylitol, various inorganic salts, and anhydrous α-D-glucose, mimicking extracellular fluid components, can reduce organ damage caused by ischemia-reperfusion. The addition of xylitol, synergistically with tetramethylpyrazine and furosemide, effectively reduces the risk of pulmonary edema. This invention provides a lung perfusion preservation solution that can be used at room temperature, significantly reducing costs, alleviating the economic burden on lung transplant patients, and enabling the domestic production of lung perfusion preservation solutions.

[0020] The perfusion preservation solution of this invention is superior to imported Steen solution in reducing pulmonary edema. For infected donor lung tissue, the novel perfusion preservation solution of this invention can be used for maintenance before transplantation, reducing the risk of infection after lung transplantation and reducing the patient's medical risks and treatment costs. Attached Figure Description

[0021] Figure 1 The oxygenation index of isolated lungs after perfusion using the lung perfusion preservation solution prepared in Example 1 of this invention, which can be used at room temperature;

[0022] Figure 2 The compliance diagram shows the lung perfusion after in vitro perfusion using the lung perfusion preservation solution prepared in Example 1 of this invention, which can be used at room temperature.

[0023] Figure 3 To obtain tidal volume and airway peak pressure maps after in vitro lung perfusion using the lung ex vivo perfusion preservation solution prepared in Example 1 of this invention, which can be used at room temperature. Detailed Implementation

[0024] To make the technical problems, technical solutions and advantages of the present invention clearer, a detailed description will be given below in conjunction with the accompanying drawings and specific embodiments.

[0025] Unless otherwise specified, all materials and reagents used in this invention are commercially available.

[0026] This invention provides a lung ex vivo perfusion preservation solution that can be used at room temperature and its preparation method. Specific embodiments are as follows.

[0027] Example 1

[0028] An ex vivo lung perfusion preservation solution that can be used at room temperature comprises the following components:

[0029] Dextran 40, sodium chloride, potassium chloride, calcium chloride dihydrate, sodium dihydrogen phosphate monohydrate, sodium bicarbonate, magnesium chloride, anhydrous α-D-glucose, xylitol, human serum albumin, ligustrazine, furosemide, sodium hydroxide, and sterile water. The concentrations of each substance are shown in Table 1.

[0030] The preparation method of the above-mentioned lung ex vivo perfusion preservation solution that can be used at room temperature includes:

[0031] 1) Prepare raw materials according to the concentrations of each substance in Table 1;

[0032] 2) Heat and dissolve dextran 40 and ligustrazine in sterile water for later use;

[0033] 3) Dissolve sodium chloride, potassium chloride, sodium dihydrogen phosphate monohydrate, sodium bicarbonate, magnesium chloride, anhydrous α-D-glucose, and xylitol in sterile water; titrate with sodium hydroxide to pH 7.4 and set aside; the concentration of sodium hydroxide is 1 mol / L.

[0034] 4) Mix the solution prepared in step 2) with the solution prepared in step 3), then add calcium chloride dihydrate, human serum albumin, and furosemide, adjust the volume with sterile water, and then adjust the pH to 7.4 with sodium hydroxide for later use;

[0035] 5) Filter the above liquid using a sterile filter, dispense and seal for storage at 4°C.

[0036] The ex vivo lung perfusion preservation solution prepared in Example 1 above, which can be used at room temperature, was used in animal experiments.

[0037] Pretreatment: The experimental Bama pigs were injected intramuscularly with 2 mL of atropine. 15 min later, 2 mL of salbutamol was injected. After anesthesia, the pigs were weighed (30.6 kg). An indwelling needle was inserted into the marginal ear vein and propofol was continuously injected. The monitor was connected, the endotracheal tube was intubated, and mechanical ventilation was implemented.

[0038] Lung perfusion in vivo: Open the chest, insert an irrigation tube through the pulmonary artery incision, cut open the left atrial appendage, and inject 2L of low potassium dextran solution through the irrigation tube to irrigate the lungs. The irrigation fluid can be seen flowing out from the opening of the left atrial appendage.

[0039] Lung harvesting: Clamp the trachea, cut the aorta and superior and inferior vena cava, cut the trachea and esophagus, carefully remove the whole heart and lungs, cut open the left atrium, the proximal aorta and the root of the pulmonary artery along the left atrial appendage incision, separate the heart and lungs, and obtain the ex vivo lungs.

[0040] Retrograde lavage of isolated lungs: 1 L of low-potassium dextran solution was injected into the pulmonary vein for retrograde lavage, and the lavage fluid was observed flowing out from the pulmonary artery incision. Then, the isolated lung perfusion preservation solution prepared in Example 1, which can be used at room temperature, was used for isolated lung perfusion repair, which could be maintained for 24 hours without lung edema.

[0041] During perfusion, blood gas analysis was performed on the perfusion fluid flowing out via the pulmonary vein at the start of perfusion and every 3 hours. Before the analysis, the oxygen concentration fraction in the inhaled air was set to 100%. Pulmonary venous oxygen pressure (PaO2) data were obtained at each time point, and the oxygenation index (PO2 / FiO2) was calculated. Figure 1 As shown.

[0042] Depend on Figure 1 It can be seen that during the entire perfusion process, the oxygenation index first decreased slightly and then continued to rise, eventually reaching 362 mmHg, which is much higher than the data at the initial perfusion. This proves that the lung ex vivo perfusion preservation solution of the present invention, which can be used at room temperature, will improve the oxygenation capacity of the ex vivo lung to a certain extent and has a certain repair ability on the ex vivo lung after perfusion.

[0043] During perfusion, the ventilator dynamically monitors the lungs' tidal volume, positive end-expiratory pressure, peak airway pressure, and compliance in real time. Data from the ventilator at 0 hours of perfusion and every hour is recorded. Figure 2 As shown. By Figure 2 It can be seen that during perfusion, the overall compliance of the lungs showed an upward trend, proving that the lung tissue has good elasticity, low airway resistance, and reduced pulmonary edema.

[0044] During perfusion, the ventilator dynamically monitors the tidal volume and peak airway pressure of the lungs in real time. Data from 0 hours of perfusion and every hour is recorded from the ventilator's real-time data. Figure 3 As shown. From Figure 3 As can be seen, during perfusion, tidal volume and peak airway pressure remained relatively stable, with a slight increase in tidal volume, indicating improved lung function. The overall increase in tidal volume throughout the perfusion process suggests a reduction in pulmonary edema and improved lung function.

[0045] Example 2

[0046] An ex vivo lung perfusion preservation solution that can be used at room temperature comprises the following components:

[0047] Dextran 40, sodium chloride, potassium chloride, calcium chloride dihydrate, sodium dihydrogen phosphate monohydrate, sodium bicarbonate, magnesium chloride, anhydrous α-D-glucose, xylitol, human serum albumin, ligustrazine, furosemide, sodium hydroxide, and sterile water. The concentrations of each substance are shown in Table 1.

[0048] The remaining conditions are the same as in Example 1.

[0049] The lung ex vivo perfusion preservation solution prepared in this embodiment, which can be used at room temperature, can be used for lung ex vivo perfusion repair for 22 hours without lung edema.

[0050] Example 3

[0051] An ex vivo lung perfusion preservation solution that can be used at room temperature comprises the following components:

[0052] Dextran 40, sodium chloride, potassium chloride, calcium chloride dihydrate, sodium dihydrogen phosphate monohydrate, sodium bicarbonate, magnesium chloride, anhydrous α-D-glucose, xylitol, human serum albumin, ligustrazine, furosemide, sodium hydroxide, and sterile water. The concentrations of each substance are shown in Table 1.

[0053] The remaining conditions are the same as in Example 1.

[0054] The lung ex vivo perfusion preservation solution prepared in this embodiment, which can be used at room temperature, can be used for lung ex vivo perfusion repair for 23 hours without lung edema.

[0055] Example 4

[0056] An ex vivo lung perfusion preservation solution that can be used at room temperature comprises the following components:

[0057] Dextran 40, sodium chloride, potassium chloride, calcium chloride dihydrate, sodium dihydrogen phosphate monohydrate, sodium bicarbonate, magnesium chloride, anhydrous α-D-glucose, xylitol, human serum albumin, ligustrazine, furosemide, sodium hydroxide, and sterile water. The concentrations of each substance are shown in Table 1.

[0058] The remaining conditions are the same as in Example 1.

[0059] The lung ex vivo perfusion preservation solution prepared in this embodiment, which can be used at room temperature, was used for lung ex vivo perfusion repair, which can maintain the condition for 19 hours without lung edema.

[0060] Table 1

[0061]

[0062]

[0063] To further illustrate the beneficial effects of the present invention, the following comparative examples were constructed.

[0064] Comparative Example 1

[0065] In this comparative perfusion preservation solution, xylitol was omitted, and the other conditions were the same as in Example 1.

[0066] The perfusion preservation solution prepared in this comparative example was used for ex vivo lung perfusion repair, which could be maintained for 13 hours without lung edema.

[0067] Comparative Example 2

[0068] In this comparative perfusion preservation solution, xylitol was replaced with an equal amount of sorbitol, and the other conditions were the same as in Example 1.

[0069] The perfusion preservation solution prepared in this comparative example was used for ex vivo lung perfusion repair, which could be maintained for 12 hours without lung edema.

[0070] Comparative Example 3

[0071] In this comparative perfusion preservation solution, tetramethylpyrazine was omitted, and the other conditions were the same as in Example 1.

[0072] The perfusion preservation solution prepared in this comparative example was used for ex vivo lung perfusion repair, which could be maintained for 11 hours without lung edema.

[0073] Comparative Example 4

[0074] In this comparative perfusion preservation solution, furosemide was omitted, and the other conditions were the same as in Example 1.

[0075] The perfusion preservation solution prepared in this comparative example was used for ex vivo lung perfusion repair, which could be maintained for 13 hours without lung edema.

[0076] Comparative Example 5

[0077] In this comparative perfusion preservation solution, the amount of potassium chloride used was 0.3 g / L, and the other conditions were the same as in Example 1.

[0078] The perfusion preservation solution prepared in this comparative example was used for ex vivo lung perfusion repair, which could be maintained for 16 hours without lung edema.

[0079] In summary, the lung perfusion preservation solution prepared by this invention, which can be used at room temperature, can preserve isolated lungs for a longer period of time through the interaction of human serum albumin, inorganic salts, tetramethylpyrazine, furosemide, xylitol, etc., and effectively reduce the risk of pulmonary edema, greatly reducing costs and alleviating the economic burden on lung transplant patients.

[0080] The above description is a preferred embodiment of the present invention. For those skilled in the art, any improvements and modifications made without departing from the principles of the present invention should also be considered within the scope of protection of the present invention.

Claims

1. A lung ex vivo perfusion preservation solution that can be used at room temperature, characterized in that, It consists of the following components: Dextran 40, sodium chloride, potassium chloride, calcium chloride dihydrate, sodium dihydrogen phosphate monohydrate, sodium bicarbonate, magnesium chloride, anhydrous α-D-glucose, xylitol, human serum albumin, ligustrazine, furosemide, sodium hydroxide, and sterile water; The concentration of dextran 40 is 4-6 g / L; the concentration of sodium chloride is 4-6 g / L; the concentration of potassium chloride is 0.4-0.6 g / L; the concentration of calcium chloride dihydrate is 0.2-0.25 g / L; the concentration of sodium dihydrogen phosphate monohydrate is 0.1-0.2 g / L; the concentration of sodium bicarbonate is 0.8-1.4 g / L; the concentration of magnesium chloride is 0.1-0.15 g / L; the concentration of anhydrous α-D-glucose is 1.5-2 g / L; the concentration of xylitol is 0.4-0.6 g / L; the concentration of human serum albumin is 66-72 g / L; the concentration of tetramethylpyrazine is 0.1-0.17 g / L; and the concentration of furosemide is 0.01-0.03 g / L.

2. The lung ex vivo perfusion preservation solution that can be used at room temperature according to claim 1, characterized in that, The concentration of dextran 40 is 5 g / L; the concentration of sodium chloride is 5 g / L; the concentration of potassium chloride is 0.54 g / L; the concentration of calcium chloride dihydrate is 0.22 g / L; the concentration of sodium dihydrogen phosphate monohydrate is 0.14 g / L; the concentration of sodium bicarbonate is 1.26 g / L; the concentration of magnesium chloride is 0.11 g / L; the concentration of anhydrous α-D-glucose is 1.8 g / L; the concentration of xylitol is 0.4 g / L; the concentration of human serum albumin is 70 g / L; the concentration of tetramethylpyrazine is 0.14 g / L; and the concentration of furosemide is 0.02 g / L.

3. The lung ex vivo perfusion preservation solution that can be used at room temperature according to claim 2, characterized in that, The pH of the ex vivo lung perfusion preservation solution, which can be used at room temperature, is 7.2-7.

4.

4. The method for preparing the ex vivo lung perfusion preservation solution as described in any one of claims 1-3, characterized in that, include: 1) Provide raw materials for the lung ex vivo perfusion preservation fluid components that can be used at room temperature according to any one of claims 1-3; 2) Heat and dissolve dextran 40 and ligustrazine in sterile water for later use; 3) Dissolve sodium chloride, potassium chloride, sodium dihydrogen phosphate monohydrate, sodium bicarbonate, magnesium chloride, anhydrous α-D-glucose, and xylitol in sterile water; titrate with sodium hydroxide to pH 7.4, and set aside. 4) Mix the solution prepared in step 2) with the solution prepared in step 3), then add calcium chloride dihydrate, human serum albumin, and furosemide, and adjust the volume with sterile water. Then adjust the pH to 7.2-7.4 with sodium hydroxide and set aside. 5) Filter the above liquid using a sterile filter, dispense and seal for storage at 4°C.

5. The method for preparing the lung ex vivo perfusion preservation solution that can be used at room temperature according to claim 4, characterized in that, The concentration of sodium hydroxide is 1 mol / L.

6. The use of the lung perfusion preservation solution that can be used at room temperature as described in any one of claims 1 to 3 in the preservation of isolated lungs.

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