A perfusate for donation after cardiac death organ procurement and uses thereof

Abstract

The application provides a kind of for heart death after organ acquisition perfusion fluid and its application, belong to medical technical field.The application is used for heart death after organ acquisition perfusion fluid, including the following components: thrombolytic drug, vasodilator, albumin, dexamethasone sodium phosphate, low molecular weight heparin sodium and lactated Ringer's solution.The organ acquisition perfusion fluid of the application can effectively remove the thrombus formed by coagulation system activation, reduce vasospasm caused by warm ischemic injury and tissue cell edema in the process of perfusion, which can effectively improve the quality and function of the acquired organ, increase the utilization rate of the organ, and has clinical transformation application prospect.

Description

Technical Field

[0001] This invention relates to the field of medical technology, and in particular to an irrigation fluid for obtaining donated organs after cardiac death and its application. Background Technology

[0002] Organ transplantation is the only effective treatment for patients with end-stage organ failure. According to statistics from the World Health Organization, approximately 130,000 people worldwide underwent solid organ transplants in 2020, meeting the needs of only ≤10% of end-stage organ failure patients awaiting transplants. Most patients are removed from the waiting list due to complete organ failure and severe complications. With the rapid development of medical and health technologies, patients whose lives are saved through organ transplantation can effectively regain normal physiological functions, significantly improving their quality of life and mental and psychological well-being. However, the supply of living donors or brain-dead donors is extremely limited, far from meeting the current demand for organ transplants, and the growth rate of potential donor organ sources is limited.

[0003] According to 2015 data on global causes of death and potential organ donation, cardiac death is the leading cause of death worldwide, with 42.13% (approximately 24 million people) becoming potential organ donors after cardiac death—about 3.8 times the rate of potential brain-dead donors. Although cardiac death donation is considered the primary method for expanding the source of donor organs, the utilization rate of these organs varies significantly across countries, ranging from approximately 17.1% to 80.0%. This is mainly because these donated organs inevitably undergo warm ischemia-reperfusion injury, resulting in a decline in organ quality and a significantly increased incidence of delayed functional recovery, primary non-function, and acute rejection after transplantation. Due to concerns about organ transplant outcomes, most potential cardiac death donors terminate their donations, and some acquired organs are even discarded after evaluation as unsuitable for clinical transplantation. If these cardiac death-donated organs could be effectively utilized, it would significantly address the urgent need for organ transplantation.

[0004] Therefore, scientists in the field of organ transplantation worldwide are conducting in-depth research on how to effectively protect, repair, and utilize organs donated after cardiac death. They have carried out numerous animal experiments and clinical studies, and developed different types of organ preservation solutions and mechanical perfusion repair platforms to increase the clinical application rate of organs donated after cardiac death. Although significant scientific research results have been achieved, the shortage of organ supply and demand has not been reversed, and the results are not ideal. Both the improvement of organ preservation solutions and the optimization of mechanical perfusion repair platforms are mainly applied to preservation techniques from organ retrieval to organ implantation, often neglecting the pathophysiological changes within donor organs after cardiac death and failing to implement effective organ maintenance measures from the very beginning of the organ retrieval stage.

[0005] Currently, the conventional method for organ retrieval involves direct irrigation with organ preservation solutions (such as UW solution, HTK solution, hypertonic citrate purine solution, etc.). After irrigation, the organs are either statically cryopreserved by immersing them in the preservation solution or dynamically preserved or repaired using an organ mechanical perfusion platform. However, with the cessation of heartbeat and the slowing or even stopping of blood flow, the coagulation system is activated, leading to the formation of numerous thrombi throughout the vascular system and their blockage of blood vessels. Simultaneously, tissue ischemia and hypoxia cause dysfunction in vasoconstriction and vasodilation within the organ, inducing persistent vasoconstriction and complete occlusion of the lumen. While conventional infusion of organ preservation solutions can flush out some thrombi in larger vessels, thrombi in the microcirculation, due to the smaller vessel diameter, tend to accumulate significantly and are often difficult to flush out completely. The presence of numerous residual microthrombi after organ retrieval results in uneven distribution of the preservation solution within the organ, preventing the organ from achieving the desired effect during static cryopreservation. Blockage of the capillary network by thrombi is a major reason why mechanical perfusion methods have limited effectiveness in repairing organs after cardiac death. Similarly, it can aggravate post-transplant reperfusion damage, leading to delayed or irreversible transplant function.

[0006] The accumulation of thrombi and vasospasm result in extremely poor irrigation with organ preservation solutions, making it difficult for the solution to distribute evenly throughout the organ. This fails to mitigate cold ischemia damage during cryopreservation, significantly reducing the quality and preservation effectiveness of the harvested organs. Organs subjected to combined warm and cold ischemia injuries will suffer severe ischemia-reperfusion injury after transplantation, inevitably leading to serious complications. Organs with a large number of residual microthrombi are often deemed unsuitable for transplantation after pathological evaluation because, upon reperfusion, new thrombi will rapidly form on top of existing ones, severely hindering the recovery of organ microcirculation perfusion function.

[0007] Therefore, employing effective irrigation methods to remove intravascular thrombi and relieve vasospasm during organ procurement is fundamental and crucial for increasing the utilization rate of donated organs after cardiac death. However, there is currently a lack of effective organ irrigation fluids specifically designed for clinical use in organ procurement after cardiac death. This severely limits the acquisition, utilization, preservation, transportation, in vitro repair, and even final surgical transplantation of donated organs after cardiac death, becoming a significant obstacle to the rapid development of the organ transplantation field.

[0008] A previous Chinese patent application, CN115777692A (publication date March 14, 2023), disclosed a room-temperature mechanical lavage fluid for kidneys from cardiac death donors. However, this patent application addresses the in vitro repair of donated organs obtained after cardiac death using a room-temperature mechanical perfusion platform. It is an intervention method during organ retrieval and preservation, primarily used to repair organs with poor quality scores after pathological histological evaluation. Compared to interventions taken in the early organ retrieval stage, this is a "better late than never" approach. Therefore, providing a lavage fluid for retrieving donated organs after cardiac death is essential. Summary of the Invention

[0009] The purpose of this invention is to provide an irrigation fluid for organ retrieval after cardiac death and its application. The organ retrieval irrigation fluid of this invention can effectively remove thrombi formed by activation of the coagulation system, reduce vasospasm caused by thermal ischemia injury, and alleviate tissue edema during irrigation. This irrigation fluid can effectively improve the quality and function of the retrieved organs, increase organ utilization, and has promising prospects for clinical translational applications.

[0010] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0011] This invention provides an irrigation fluid for obtaining donated organs after cardiac death, comprising the following components: thrombolytic drugs, vasodilators, albumin, dexamethasone sodium phosphate, low molecular weight heparin sodium, and lactated Ringer's solution.

[0012] Preferably, the thrombolytic drug includes any one of streptokinase, urokinase, alteplase, and plasmin.

[0013] Preferably, the vasodilator includes any one of fasudil hydrochloride, urapidil hydrochloride, and alprostadil.

[0014] Preferably, in the irrigation solution, sodium lactate Ringer's solution is used as the solvent, the concentration of streptokinase is 10,000-15,000 IU / mL, the concentration of urokinase is 2,000-20,000 IU / mL, the concentration of alteplase is 0.5-2 mg / mL, and the concentration of plasmin is 5-50 IU / mL.

[0015] Preferably, in the irrigation solution, sodium lactate Ringer's solution is used as the solvent, the concentration of fasudil hydrochloride is 0.05-10 mg / mL, the concentration of urapidil hydrochloride is 0.05-8 mg / mL, and the concentration of alprostadil is 0.05-10 μg / mL.

[0016] Preferably, in the irrigation solution, sodium lactate Ringer's solution is used as the solvent, the concentration of albumin is 100-180 g / L, the concentration of dexamethasone sodium phosphate is 5-20 mg / L, and the concentration of low molecular weight heparin sodium is 12000-13000 IU / L.

[0017] The present invention also provides an application of the above-mentioned lavage fluid for obtaining donated organs after cardiac death, wherein lavage is performed during organ retrieval when the thermal ischemia injury time after cardiac death is 5 min to 90 min.

[0018] This invention provides an irrigation solution for procuring donated organs after cardiac death and its application. Compared to standard donor organ procuring, organs donated after cardiac death suffer varying degrees of thermal ischemia-reperfusion injury. If crystalloid solutions such as physiological saline, lactated Ringer's solution, or compound electrolyte solutions are used directly for irrigation, or as drug solvents, tissue edema often occurs. This tissue edema compresses surrounding capillaries, exacerbating vascular occlusion. The organ irrigation solution of this invention, in addition to components such as lactated Ringer's solution, also contains albumin as a colloidal component and the glucocorticoid dexamethasone sodium phosphate, to prevent tissue damage and reduce tissue edema during irrigation.

[0019] The organ retrieval lavage fluid of this invention can not only effectively remove diffuse microthrombus accumulation in donated organs after cardiac death and alleviate ischemic spasm of capillaries within the organs, but also reduce tissue edema during the organ retrieval lavage process. This invention solves the current problem of lacking a lavage fluid specifically for retrieval of donated organs after cardiac death. This lavage fluid can more effectively improve the quality and function of retrieved organs, increase organ utilization, and improve the survival rate of transplanted organs. Attached Figure Description

[0020] Figure 1 The images show the histopathological examination results of the liver and kidney obtained using organ preservation solution, irrigation solution from Example 1, Example 2, and Example 3, respectively. Detailed Implementation

[0021] This invention provides an irrigation fluid for obtaining donated organs after cardiac death, comprising the following components: thrombolytic drugs, vasodilators, albumin, dexamethasone sodium phosphate, low molecular weight heparin sodium, and lactated Ringer's solution.

[0022] In this invention, the thrombolytic drug preferably includes any one of streptokinase, urokinase, alteplase, and plasmin, and is more preferably plasmin.

[0023] In this invention, the vasodilator preferably includes any one of fasudil hydrochloride, urapidil hydrochloride, and alprostadil, and more preferably urapidil hydrochloride.

[0024] In this invention, the irrigation solution uses sodium lactate Ringer's solution as a solvent, and the concentration of the streptokinase is preferably 10,000 to 15,000 IU / mL, more preferably 12,000 to 15,000 IU / mL, and even more preferably 14,000 IU / mL.

[0025] In this invention, the irrigation solution uses sodium lactate Ringer's solution as a solvent, and the concentration of urokinase is preferably 2000-20000 IU / mL, more preferably 4000-20000 IU / mL, and even more preferably 10000 IU / mL.

[0026] In this invention, the irrigation solution uses sodium lactate Ringer's solution as the solvent, and the concentration of alteplase is preferably 0.5-2 mg / mL, more preferably 1-2 mg / mL, and even more preferably 1.5 mg / mL.

[0027] In this invention, the irrigation solution uses sodium lactate Ringer's solution as the solvent, and the concentration of the plasmin is preferably 5-50 IU / mL, more preferably 20-50 IU / mL, and even more preferably 30 IU / mL.

[0028] In this invention, the irrigation solution uses sodium lactate Ringer's solution as a solvent, and the concentration of fasudil hydrochloride is preferably 0.05-10 mg / mL, more preferably 0.1-10 mg / mL, and even more preferably 8 mg / mL.

[0029] In this invention, the irrigation solution uses sodium lactate Ringer's solution as the solvent, and the concentration of urapidil hydrochloride is preferably 0.05-8 mg / mL, more preferably 0.1-8 mg / mL, and even more preferably 4 mg / mL.

[0030] In this invention, the irrigation solution uses sodium lactate Ringer's solution as the solvent, and the concentration of alprostadil is preferably 0.05-10 μg / mL, more preferably 0.1-10 μg / mL, and even more preferably 8 μg / mL.

[0031] In this invention, sodium lactate Ringer's solution is used as the solvent in the irrigation fluid, and the concentration of albumin is preferably 100-180 g / L, more preferably 120-150 g / L, and even more preferably 120 g / L.

[0032] In this invention, the irrigation solution uses sodium lactate Ringer's solution as the solvent, and the concentration of dexamethasone sodium phosphate is preferably 5-20 mg / L, more preferably 10-18 mg / L, and even more preferably 10 mg / L.

[0033] In this invention, the irrigation solution uses sodium lactate Ringer's solution as a solvent, and the concentration of the low molecular weight heparin sodium is 12,000 to 13,000 IU / L, more preferably 12,500 IU / L.

[0034] The present invention also provides an application of the above-mentioned lavage fluid for obtaining donated organs after cardiac death, wherein lavage is performed during organ retrieval when the thermal ischemia injury time after cardiac death is 5 min to 90 min.

[0035] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0036] Example 1

[0037] This embodiment provides an irrigation fluid for obtaining donated organs after cardiac death, and the specific preparation process is as follows:

[0038] Prepare 1000 mL of irrigation solution using sodium lactate Ringer's solution as the solvent. The solution contains plasmin at a concentration of 30 IU / mL, fasudil hydrochloride at a concentration of 8 mg / mL, albumin at a concentration of 100 g / L, dexamethasone sodium phosphate at a concentration of 10 mg / L, and low molecular weight heparin sodium at a concentration of 12500 IU / L.

[0039] Example 2

[0040] This embodiment provides an irrigation fluid for obtaining donated organs after cardiac death, and the specific preparation process is as follows:

[0041] Prepare 1000 mL of irrigation solution using sodium lactate Ringer's solution as the solvent. The solution contains plasmin at a concentration of 30 IU / mL, urapidil hydrochloride at a concentration of 4 mg / mL, albumin at a concentration of 120 g / L, dexamethasone sodium phosphate at a concentration of 15 mg / L, and low molecular weight heparin sodium at a concentration of 12500 IU / L.

[0042] Example 3

[0043] This embodiment provides an irrigation fluid for obtaining donated organs after cardiac death, and the specific preparation process is as follows:

[0044] Prepare 1000 mL of irrigation solution using sodium lactate Ringer's solution as the solvent. The solution contains plasmin at a concentration of 50 IU / mL, alprostadil at a concentration of 8 μg / mL, albumin at a concentration of 130 g / L, dexamethasone sodium phosphate at a concentration of 10 mg / L, and low molecular weight heparin sodium at a concentration of 12500 IU / L.

[0045] Experimental Example 1

[0046] This experimental case establishes a model of organ donation after cardiac death to verify the effect of the lavage fluids used in Examples 1-3 on the lavage of donated organs after cardiac death. The specific process is as follows:

[0047] (1) Organ donation model after cardiac death

[0048] Four SPF-grade male SD rats aged 6–8 weeks were selected and fasted for 12 hours before the procedure. After anesthesia with 1% sodium pentobarbital via intraperitoneal injection, the abdominal hair was shaved and the rats were fixed on the operating table. Cardiac arrest was induced by bilateral thoracic incision. The peritoneal temperature was maintained at 36.5–37°C using a constant temperature heating pad, and the warm ischemia time was calculated to be 90 min.

[0049] (2) Organ procurement

[0050] One rat was treated with organ preservation solution (UW solution, University of Wisconsin, USA), and the irrigation solutions of Examples 1, 2, and 3, respectively. The liver and kidneys were irrigated by cannulating the abdominal aorta and portal vein of the rat with the preservation solution / irrigation solution. An opening was cut in the superior and inferior vena cava of the liver. After clear liquid flowed out, organ preservation solution at 4°C was infused and the rat was immersed in organ preservation solution for low-temperature preservation.

[0051] (3) Examination of irrigation organs

[0052] The histopathological examination results of the liver and kidneys obtained 90 minutes after cardiac death following lavage with organ preservation solution and lavage solutions from Examples 1-3 are as follows: Figure 1 As shown.

[0053] As can be seen from histopathological HE staining, after perfusion of the liver and kidneys in groups 1-3 90 minutes after cardiac death, there was no significant accumulation of red blood cells in the capillaries, and the tissue structure was intact, reflecting good organ perfusion and structural condition. However, after perfusion with organ preservation fluid, a large amount of red blood cell accumulation and thrombus obstruction of blood vessels remained. Figure 1 (As indicated by the black arrow in the middle).

[0054] 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 application for obtaining lavage fluid from donated organs after cardiac death, characterized in that, Irrigation was performed during organ retrieval, with a post-cardiac death warm ischemia injury time of 5 to 90 minutes; the donated organ was the liver. The irrigation fluid used for obtaining organs from donors after cardiac death consists of the following components: thrombolytic drugs, vasodilators, albumin, dexamethasone sodium phosphate, low molecular weight heparin sodium, and lactated Ringer's solution. The thrombolytic drugs include any one of streptokinase, urokinase, alteplase, and plasmin. The vasodilator includes any one of fasudil hydrochloride, urapidil hydrochloride, and alprostadil. The concentration of the streptokinase is 10,000-15,000 IU / mL, the concentration of the urokinase is 2,000-20,000 IU / mL, the concentration of the alteplase is 0.5-2 mg / mL, and the concentration of the plasmin is 5-50 IU / mL. The concentration of fasudil hydrochloride is 0.05~10 mg / mL, the concentration of urapidil hydrochloride is 0.05~8 mg / mL, and the concentration of alprostadil is 0.05~10 μg / mL; The albumin concentration is 100~180g / L, the dexamethasone sodium phosphate concentration is 5~20mg / L, and the low molecular weight heparin sodium concentration is 12000~13000IU / L.

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