TWI891029B - Treatment for acute organ injury using cd39, recombinant cd39 - Google Patents

Abstract

The present invention pertains to the use or methods for treating organ injury, in particular acute organ injury, such as acute kidney injury (AKI) using CD39, human recombinant CD39 and variants thereof.

Description

Using CD39 and recombinant CD39 for the treatment of acute organ damage

The present disclosure generally relates to methods of using CD39, human recombinant CD39, and variants thereof (e.g., recombinant CD39 variants having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-21) for treating organ injury, particularly acute organ injury (e.g., acute renal injury (AKI)).

Acute kidney injury (AKI) is an asymptomatic condition characterized by a sudden deterioration in kidney function, sometimes accompanied by a decrease in urine output or even a complete loss of urine output. Most patients experience this syndrome during hospitalization, especially those who are critically ill and undergoing major surgery. Recovery from AKI is possible; however, in some patients, kidney function will continue to decline and remain impaired long-term. Some patients will require at least short-term dialysis or will develop severe kidney impairment and remain dialysis-dependent, requiring a kidney transplant.

Furthermore, during sepsis, the host's dysregulated response to infection can lead to life-threatening multi-organ dysfunction. Sepsis is a leading cause of death and a major public health problem affecting millions of patients worldwide each year. Approximately 90% of sepsis cases are caused by bacterial infection, and 40%-60% of septic patients develop acute kidney injury (AKI). AKI is a common complication of sepsis, affecting 28% of patients. Acute kidney injury (AKI) in patients with sepsis is associated with a significant increase in mortality, up to 65%, and survivors are at risk for chronic kidney disease (CKD) and end-stage renal disease, resulting in significant health and economic burdens for both patients and society. Sepsis-associated acute kidney injury (SA-AKI) is a multifactorial syndrome characterized by the simultaneous occurrence of inflammatory, renal toxic, and ischemic damage, along with other pathophysiological responses, leading to rapid kidney damage. Over the past 30 years, a significant number of potential drug targets have emerged, leading to the development of new therapies for the treatment of sepsis and AKI. Attempts have primarily focused on improving hemodynamics, reducing oxidative stress, and blocking the hyperinflammatory response; however, all of these have failed in clinical trials. Treatment of SA-AKI patients relies primarily on antibiotics and supportive care, including fluid resuscitation, vasoactive agents, and renal replacement therapy.

AKI is also a common and serious complication of cardiopulmonary bypass surgery (CPBS) and cardiac surgery-associated AKI (CSA-AKI). AKI is new or worsening renal failure characterized by a relatively sudden drop in glomerular filtration rate (GFR), often accompanied by decreased urine output. AKI typically occurs within the first 7 days after the initial injury, most commonly following a transient hypotensive episode of any cause, but may also occur in response to renal toxins or radioactive contrast agents. The clinical incidence of AKI is 3%-18% of all hospitalized adult patients worldwide and is more common in the setting of complex surgery. By definition, AKI occurs in up to 15%-40% of adults after CPBS. Severe AKI requires renal replacement therapy in 1%-5% of cases and is associated with a mortality rate as high as 70%.

Several lines of evidence suggest that converting extracellular ATP to adenosine has potential benefits in treating sepsis and associated organ damage. In an animal model of cecal ligation and puncture (CLP)-induced sepsis, treatment with adenosine (ATPase), an ATPase that promotes the enzymatic breakdown of extracellular ATP, reduced extracellular ATP concentrations in peritoneal lavage and significantly reduced mortality.

A clinical study using alkaline phosphatase (ALP) in patients with SA-AKI showed that AP improved renal function, as determined by a composite endpoint of creatinine clearance, RRT requirement, and RRT duration. Alkaline phosphatases are endogenous enzymes that detoxify by dephosphorylating endotoxins and extracellular ATP. In a relatively large clinical study of patients with SA-AKI (N = 301), results suggested a potential mortality benefit with AP treatment compared with placebo, although the study did not meet the primary endpoint of improvement in short-term renal function (measured by area corrected for endogenous creatinine clearance from day 1 to day 7).

There are currently no approved drug therapies to treat or prevent AKI or SA-AKI.

We have now developed novel treatments using recombinant CD39 and its variants, as defined herein, that are safe, effective, and provide sustained responses in subjects with common acute organ injury and AKI, particularly AKI such as sepsis-associated acute renal injury (SA-AKI) or cardiac surgery-associated AKI (CSA-AKI). These new treatments address a long-standing need for safe and effective AKI treatments for clinicians and patients. Therapeutic approaches that utilize the mechanism by which recombinant CD39 hydrolyzes extracellular ATP and ADP to AMP to enhance the conversion of extracellular ATP to adenosine represent a therapeutic strategy to mitigate and inhibit common acute organ injury. Due to efficient hydrolysis of ATP and ADP, recombinant CD39, as defined herein, dose-dependently inhibits ATP/LPS-induced IL-1β secretion and ADP- and thrombin-induced platelet aggregation. The inhibition of thrombin-induced platelet aggregation is attributed to the release of ADP from platelet dense granules via activation of protease-activated receptor 4 (PAR4) by thrombin inducers. Furthermore, recombinant CD39, as defined herein, inhibits ATP/LPS-induced IL-1β release in a concentration-dependent manner. Such treatment exhibits anti-inflammatory, anticoagulant, and tissue-protective effects, potentially improving renal function and reducing mortality in subjects with AKI (e.g., SA-AKI or CSA-AKI).

In one embodiment, recombinant CD39 is provided, comprising an amino acid sequence of SEQ ID NOs: 1 to 21, wherein the recombinant CD39 is administered to a subject in need thereof at a dose of about 0.1 mg/kg to about 10 mg/kg body weight (mg/kg).

In a preferred embodiment, recombinant CD39 designated as CD39* is provided. Specifically, CD39* comprises the amino acid sequence of SEQ ID NO: 21, and wherein the recombinant CD39* is administered to a subject in need thereof at a dose of about 1 mg/kg to about 5 mg/kg.

In one embodiment, the route of administration is intravenous (IV), with IV infusion of recombinant CD39 according to the embodiments described herein.

In another example, a suitable regimen for recombinant CD39 (as defined herein) is a daily regimen.

In some embodiments, recombinant CD39 (as defined herein, preferably CD39*) can be administered to a subject at a dose of about 0.1 mg/kg to about 10 mg/kg, for example, at a dose of about 1 mg/kg, 2 mg/kg, 3 mg/kg, 4 mg/kg, or about 5 mg/kg delivered IV.

In yet another specific embodiment, a dose of about 4 mg/kg of recombinant CD39 (preferably CD39* as defined herein) is administered IV over a 2-hour period.

In yet another specific embodiment, a dose of about 5 mg/kg of recombinant CD39 (preferably CD39* as defined herein) is administered IV over a 2-hour period.

Within hours of the first diagnosis of AKI or sepsis in a subject, recombinant CD39 (preferably CD39* as defined herein) can be administered to the subject daily by infusion, e.g., i.v., at a dose of about 4-5 mg/kg.

Recombinant CD39 (as defined herein, preferably CD39*) can be administered to the subject by infusion, such as IV, at a dose of about 4-5 mg/kg, starting from the time of suspected or confirmed AKI diagnosis in the subject.

Recombinant CD39 (as defined herein, preferably CD39*) can be administered to the subject by infusion, e.g., IV, at a dose of about 4-5 mg/kg within about 48 hours or less (e.g., within 12 hours, within 24 hours, or within 36 hours, for example) after the subject is first diagnosed with AKI.

In some embodiments, a treatment regimen may last from a few hours to a day to several weeks, as determined by a competent caregiver (treating physician).

In one embodiment, the present invention comprises administering recombinant CD39 (as defined herein, preferably CD39*) to a subject with AKI (e.g., SA-AKI or CSA-AKI) at a dose ranging from about 0.1 mg/kg to about 10 mg/kg per treatment, preferably from 1 mg/kg to 5 mg/kg per treatment, and preferably about 4-5 mg/kg per treatment. In one embodiment, the subject receives 4 or 5 mg/kg per treatment. In one embodiment, the subject with AKI (e.g., SA-AKI or CSA-AKI) receives a single treatment within about 48 hours of the initial diagnosis of AKI (e.g., SA-AKI or CSA-AKI).

In some embodiments, recombinant CD39 (as defined herein, preferably CD39*) is administered in combination with one or more additional agents. In some embodiments, the one or more additional agents are steroids, metalloproteinase inhibitors, serine protease inhibitors, local anesthetic creams, proliferative factor inhibitors, antinausea agents, or antibiotics.

In another aspect, the present disclosure provides novel dosing regimens for recombinant CD39 (as defined herein, preferably CD39*), which can be used in methods for treating or preventing AKI (e.g., SA-AKI or CSA-AKI).

In another aspect, the present disclosure provides novel dosing regimens for recombinant CD39 (preferably CD39* as defined herein), which can be used in methods for treating or preventing sepsis.

In another aspect, the present disclosure provides novel dosing regimens for recombinant CD39 (as defined herein, preferably CD39*) as a medicament, wherein the recombinant CD39 is administered to a subject in need thereof at a dose of about 0.1 mg/kg to about 10 mg/kg, for example, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg, or about 5 mg/kg. In yet another aspect, the present disclosure provides novel therapeutic methods using recombinant CD39 (as defined herein, preferably CD39*), wherein the recombinant CD39 is administered to a subject in need thereof at a dose of about 0.1 mg/kg to about 10 mg/kg, for example, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg, or about 5 mg/kg.

For the purposes of interpreting this specification, the following definitions shall apply, and where appropriate, terms used in the singular shall include the plural, and vice versa. Implementation:

A1. A recombinant CD39 for use in treating or preventing AKI in a subject in need thereof.

A2. The recombinant CD39 according to embodiment A1, wherein the AKI is severe AKI.

A3. The recombinant CD39 according to embodiment A1, wherein at least one symptom of AKI in the subject is reduced or eliminated.

A4. The recombinant CD39 according to any of the preceding embodiments, wherein the severity of AKI is reduced by at least one stage as defined by the modified AKI Network (AKIN) criteria for serum creatinine.

A5. The recombinant CD39 according to any one of embodiments A1-A4, for use in preventing AKI in a subject.

A6. The recombinant CD39 according to any one of embodiments A1-A4, for use in reducing the risk of AKI.

A7. The recombinant CD39 according to any one of embodiments A1-A4, for use in reducing the severity of AKI.

A8. The recombinant CD39 according to any of the above embodiments, wherein the incidence of severity of acute renal disease is reduced, as assessed by renal function and major adverse renal events (MAKE).

A9. The recombinant CD39 according to any of the preceding embodiments, wherein the severity of SA-AKI is reduced.

A10. The recombinant CD39 according to any of the preceding embodiments, wherein the progression of AKI, such as septic-AKI, is prevented.

A11. The recombinant CD39 according to any of the above embodiments, for use in improving major adverse renal events, such as reducing renal damage.

A12. A recombinant CD39 for use in treating or preventing sepsis.

A13. The recombinant CD39 according to embodiment A12, wherein the diagnosis of sepsis is performed according to the third edition of the international consensus definition criteria.

A14. The recombinant CD39 according to embodiment A12 or A13, wherein sepsis and septic shock (septicemia-3) are defined as follows: - suspected or confirmed infection, and - an acute increase in the Sequential Organ Failure Assessment (SOFA) score of 2 or more (excluding the renal component). The baseline SOFA score should be considered zero unless the participant is known to have pre-existing organ dysfunction (acute or chronic) prior to infection.

A15. The recombinant CD39 according to any of the above embodiments is administered at a dose of about 0.1 mg/kg to about 10 mg/kg, for example, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg or about 5 mg/kg.

A16. The recombinant CD39 according to any of the above embodiments, wherein the recombinant CD39 is administered at a dose of about 4 mg/kg of recombinant CD39.

A17. The recombinant CD39 according to any of the above embodiments, wherein the recombinant CD39 is administered at a dose of about 5 mg/kg of recombinant CD39.

A18. The recombinant CD39 according to any of the above embodiments, wherein the recombinant CD39 is administered IV, for example, administered IV over a period of about 2 hours.

A19. The recombinant CD39 according to any of the above embodiments, wherein the recombinant CD39 is administered within a few hours after the subject is first diagnosed with AKI.

A20. The recombinant CD39 according to any of the above embodiments, wherein the recombinant CD39 is administered to the subject at a dose of about 4 or 5 mg/kg daily by infusion, such as IV, starting from the time the subject is diagnosed with suspected or confirmed AKI.

A21. The recombinant CD39 according to any of the above embodiments, wherein the recombinant CD39 is administered within about 48 hours or less (e.g., within 12 hours, within 24 hours, or within 36 hours, for example) after the subject is first diagnosed with AKI.

A22. The recombinant CD39 according to any of the above embodiments, wherein the recombinant CD39 is administered within about 24 hours or as soon as possible after the first diagnosis of AKI or sepsis.

A23. The recombinant CD39 according to any of the above embodiments, wherein the recombinant CD39 is administered within a few hours after the subject is first diagnosed with sepsis.

A24. The recombinant CD39 according to any of the above embodiments, wherein the recombinant CD39 is administered to the subject at a dose of about 4 or 5 mg/kg daily by infusion, such as IV, starting from the time when the subject is diagnosed with suspected or confirmed sepsis.

A25. The recombinant CD39 according to any of the above embodiments, wherein the recombinant CD39 is administered to the subject within about 48 hours or less (e.g., within 12 hours, within 24 hours, or within 36 hours, for example) after the first diagnosis of sepsis.

A26. The recombinant CD39 according to any one of the above embodiments, wherein the recombinant CD39 has an amino acid sequence selected from the group consisting of SEQ ID NO: 1 to 21.

A27. The recombinant CD39 according to any one of the above embodiments, wherein the recombinant CD39 has the amino acid sequence of SEQ ID NO: 21.

A28. The recombinant CD39 according to any of the preceding embodiments, wherein the recombinant CD39 is administered to the subject as a single therapy.

A29. The recombinant CD39 according to any of the preceding embodiments, wherein the recombinant CD39 is co-administered with another therapeutic agent.

A30. The recombinant CD39 according to any of the above embodiments, wherein the recombinant CD39 is co-administered with an additional therapeutic agent selected from a steroid, a metalloproteinase inhibitor, a serine protease inhibitor, a local anesthetic emulsion, a proliferation factor inhibitor, an antinausea agent or an antibiotic.

A31. The recombinant CD39 according to any of the preceding embodiments, wherein the AKI is SA-AKI.

A32. The recombinant CD39 according to any of the above embodiments, wherein the AKI is cardiac surgery-associated AKI (CSA-AKI).

A33. The recombinant CD39 according to any of the preceding embodiments, wherein the recombinant CD39 is used in a method for preventing or treating one or more symptoms associated with AKI.

A34. A recombinant CD39 for use as a medicament, wherein the recombinant CD39 is administered in an amount of about 0.1 mg/kg to about 10 mg/kg, for example, in an amount of about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg or about 5 mg/kg.

A35. A recombinant CD39 for use in treating or preventing tissue damage, wherein the recombinant CD39 is administered at a dose of about 0.1 mg/kg to about 10 mg/kg, for example, at a dose of about 1 mg/kg, 2 mg/kg, about 3 mg/kg, about 4 mg/kg or about 5 mg/kg.

A36. A recombinant CD39 for use in treating or preventing tissue injury, wherein the tissue injury is acute brain injury (stroke); acute multiple organ failure; delayed graft function after kidney or other solid organ transplantation; burns; radiation injury; acute injury due to trauma and/or hypoxia, such as acute respiratory distress syndrome (ARDS) or lung injury; acute kidney injury, such as acute kidney injury secondary to thoracic surgery (e.g., aortic valve replacement, coronary artery bypass surgery) or sepsis or rhabdomyolysis or toxic effects of antibiotics or other drugs; acute myocardial injury, And wherein the recombinant CD39 is administered at a dose of about 0.1 mg/kg to about 10 mg/kg, for example, at a dose of about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg or about 5 mg/kg.

A37. A recombinant CD39 for use in the treatment of delayed graft function (DGF), acute respiratory distress syndrome (ARDS), acute myocardial infarction (AMI), traumatic brain injury (TBI)/acute ischemic stroke (AIS), ischemia-reperfusion injury (IRI), or a combination thereof, commonly referred to as multiple organ failure (MOF), wherein the recombinant CD39 is administered in an amount of about 0.1 mg/kg to about 10 mg/kg, for example, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg, or about 5 mg/kg.

A38. The recombinant CD39 for use according to embodiment A36 or A37, wherein the recombinant CD39 is administered at a dose of about 1 mg/kg of the recombinant CD39.

A39. The recombinant CD39 for use according to embodiment A36 or A37, wherein the recombinant CD39 is administered at a dose of about 2 mg/kg of the recombinant CD39.

A40. The recombinant CD39 for use according to embodiment A36 or A37, wherein the recombinant CD39 is administered at a dose of about 4 mg/kg of the recombinant CD39.

A50. The recombinant CD39 for use according to embodiment A36 or A37, wherein the recombinant CD39 is administered at a dose of about 5 mg/kg of the recombinant CD39.

A51. Recombinant CD39 for use according to embodiment A36 or A37, wherein the recombinant CD39 is administered IV, for example, is administered IV over a period of about 2 hours.

A52. The recombinant CD39 according to any one of the above embodiments, wherein the recombinant CD39 comprises the amino acid sequence of SEQ ID NO: 2.

B1. A method of treating or preventing AKI in a subject, particularly a subject in need thereof, comprising administering recombinant CD39, typically in a therapeutically effective amount.

B2. The method according to embodiment B1, wherein the AKI is severe AKI.

B3. The method of embodiment B1, wherein at least one symptom of AKI in the subject is reduced or eliminated.

B.4. The method of any one of embodiments B1-B3 above, wherein the severity of AKI is reduced by at least one stage as defined by the modified AKI Network (AKIN) criteria for serum creatinine.

B5. The method of any one of embodiments B1-B4, wherein the risk of AKI is reduced.

B6. The method of any one of embodiments B1-B4, wherein the severity of AKI is reduced.

B7. The method according to any one of embodiments B1-B4 above, wherein the incidence of severity of acute renal disease is reduced, as assessed by renal function and major adverse renal events (MAKE).

B8. The method according to any one of embodiments B1-B4 above, wherein the progression of AKI, such as septic-AKI, is arrested.

B9. The method according to any one of embodiments B1-B8 above, wherein one or more major adverse renal events are improved, such as reduced renal damage.

B10. A method of treating or preventing sepsis in a subject, particularly a subject in need thereof, comprising administering recombinant CD39, typically in a therapeutically effective amount.

B11. The method according to embodiment B10, wherein the diagnosis of sepsis is performed according to the third edition of the international consensus definition criteria.

B12. The method of embodiment B10 or B11, wherein sepsis and septic shock (septicemia-3) are defined based on the following: - suspected or confirmed infection, and - an acute increase in the Sequential Organ Failure Assessment (SOFA) score of 2 or more (excluding the renal component). The baseline SOFA score should be considered zero unless the participant is known to have pre-existing organ dysfunction (acute or chronic) prior to infection.

B13. The method according to any one of embodiments B1-B12 above, wherein the recombinant CD39 is administered at a dose of about 0.1 mg/kg to about 10 mg/kg, for example, at a dose of about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg or about 5 mg/kg.

B14. The method according to any one of embodiments B1-B13 above, wherein the recombinant CD39 is administered IV, for example, over a period of about 2 hours.

B15. The method of any one of embodiments B1-B14 above, wherein the recombinant CD39 is administered within a few hours after the subject is first diagnosed with AKI or sepsis, for example, within about 48 hours or less (e.g., within 12 hours, within 24 hours, or within 36 hours, for example) after the subject is first diagnosed with AKI.

B16. The method according to any one of embodiments B1-B15 above, wherein the recombinant CD39 is administered to the subject daily by infusion, e.g., IV, at a dose of about 4 or 5 mg/kg starting from the time the subject is diagnosed with suspected or confirmed AKI or sepsis.

B17. The method according to any one of embodiments B1-B16 above, wherein the recombinant CD39 has an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 21.

B18. The method according to any one of embodiments B1-B17 above, wherein the recombinant CD39 has the amino acid sequence of SEQ ID NO: 21.

B19. The method according to any one of embodiments B1-B18 above, wherein the recombinant CD39 is administered to the subject as a single therapy.

B20. The method according to any one of embodiments B1-B18 above, wherein the recombinant CD39 is co-administered with another therapeutic agent.

B21. The method according to embodiment B20, wherein the recombinant CD39 is co-administered with an additional therapeutic agent selected from a steroid, a metalloproteinase inhibitor, a serine protease inhibitor, a local anesthetic emulsion, a proliferation factor inhibitor, an antinausea agent, or an antibiotic.

B22. The method according to any one of embodiments B1-B21 above, wherein the AKI is SA-AKI or cardiac surgery-related AKI (CSA-AKI).

B23. A method of using recombinant CD39 for treatment, wherein the recombinant CD39 is administered at a dose of about 0.1 mg/kg to about 10 mg/kg, for example, at a dose of about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg, or about 5 mg/kg.

B24. A method of using recombinant CD39 for treating or preventing tissue damage, wherein the recombinant CD39 is administered at a dose of about 0.1 mg/kg to about 10 mg/kg, for example, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg, or about 5 mg/kg.

B25. A method of using recombinant CD39 for treating or preventing tissue injury, wherein the tissue injury is acute brain injury (stroke); acute multiple organ failure; delayed graft function after kidney or other solid organ transplantation; burns; radiation injury; acute injury due to trauma and/or hypoxia, such as acute respiratory distress syndrome (ARDS) or lung injury; acute kidney injury, such as acute kidney injury secondary to thoracic surgery (e.g., aortic valve replacement, coronary artery bypass surgery), sepsis, rhabdomyolysis, or toxic effects of antibiotics or other drugs; acute myocardial injury, And wherein the recombinant CD39 is administered at a dose of about 0.1 mg/kg to about 10 mg/kg, for example, at a dose of about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg or about 5 mg/kg.

B26. A method of using recombinant CD39 for treating delayed graft function (DGF), acute respiratory distress syndrome (ARDS), acute myocardial infarction (AMI), traumatic brain injury (TBI)/acute ischemic stroke (AIS), ischemia-reperfusion injury (IRI), or a combination thereof, commonly referred to as multiple organ failure (MOF), wherein the recombinant CD39 is administered in an amount of about 0.1 mg/kg to about 10 mg/kg, for example, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4 mg/kg, or about 5 mg/kg.

B27. The method according to any one of embodiments B1 to B26 above, wherein the recombinant CD39 comprises the amino acid sequence of SEQ ID NO: 21. Definition

As used herein, the term "CD39" refers to the human protein CD39 (cluster of differentiation 39), also known as ectonucleoside triphosphate diphosphohydrolase-1 (NTPDase 1 or Ecto-ATPDase 1). CD39 is an ectonucleotidase present on the cell surface, with a catalytic site on the extracellular surface. This ectonucleotidase catalyzes the hydrolysis of the γ- and β-phosphate residues of nucleoside triphosphates and diphosphates into their monophosphate derivatives. Specifically, human CD39 has the amino acid sequence of SEQ ID NO: 22 and/or is represented by the UniProt entry P49961.

A "CD39 variant" refers to a protein derived from human CD39 that has an amino acid sequence that is at least 60% identical to human CD39 over the entire length of human CD39. In certain embodiments, the CD39 variant consists solely of the extracellular domain of human CD39, or a portion thereof (amino acids 38 to 478 of SEQ ID NO:22). In particular, the CD39 variant has an amino acid sequence that is at least 80% identical to the extracellular domain of human CD39 over the entire length of amino acid positions 38 to 478 of human CD39. In particular, the CD39 variant is a soluble variant, i.e., a variant that lacks the transmembrane domain and its membrane anchor of human CD39. In certain embodiments, the variants have an N-terminal deletion of 30 to 50 amino acids, a C-terminal deletion of 20 to 40 amino acids, and/or a central deletion of 10 to 15 amino acids compared to human CD39. Furthermore, the variants may contain up to five point mutations compared to human CD39. Certain variants of CD39 are disclosed in WO 2020/016804. In particular, the human recombinant CD39 variants have an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 21.

In particular, "recombinant CD39" refers to a human recombinant CD39 variant having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1 to 21.

The term "about" in relation to a numerical value x means, for example, +/- 10%. When used before a numerical range or a list of numbers, the term "about" applies to each number in the series. For example, the phrase "about 1-5" should be interpreted as "about 1 - about 5," or, for example, the phrase "about 1, 2, 3, 4" should be interpreted as "about 1, about 2, about 3, about 4, etc."

The word "substantially" does not exclude "completely", for example, a composition "substantially free of" Y may completely be free of Y. If necessary, the word "substantially" may be omitted from the definitions of the present disclosure.

As used herein, a subject is "in need of" a treatment if the subject would benefit biologically, medically, or in quality of life from such treatment.

The term "pharmaceutically acceptable" refers to non-toxic materials that do not interfere with the effectiveness of the biological activity of one or more active ingredients.

The term "treatment" or "treat" is defined herein as applying or administering an ATPase according to the present invention to a subject, or applying or administering a pharmaceutical composition comprising recombinant CD39 to a subject or a tissue or cell line isolated from the subject, wherein the subject has tissue damage or symptoms associated with tissue damage, with the purpose of alleviating, reducing, or ameliorating tissue damage or any symptoms associated with tissue damage, particularly by reducing extracellular ATP levels.

"Treatment" also refers to applying or administering a pharmaceutical composition comprising recombinant CD39 to a subject, or applying or administering a pharmaceutical composition comprising an ATPase of the present invention to an isolated tissue or cell line from a subject, wherein the subject has tissue damage or symptoms associated with tissue damage, with the goal of alleviating, reducing, or ameliorating tissue damage or any symptoms associated with tissue damage.

The terms "prevent" and "preventing" refer to prophylactic and preventative treatments that focus on delaying or preventing the onset of the disease, disorder, and/or symptoms associated with the disease.

As used herein, the term "administration" or "administering" a subject compound means providing a compound of the invention and its prodrug to a subject in need of treatment. Administration "in combination" with one or more other therapeutic agents includes both simultaneous (concurrent) and consecutive administration in any order and by any route of administration.

As used herein, a "therapeutically effective amount" refers to an amount of recombinant CD39 that, when administered to a patient (e.g., a human) in a single dose or in multiple doses, is effective to treat, prevent, protect against the onset of, cure, delay, reduce the severity of, alleviate at least one symptom of a disorder or recurrence of a disorder, or prolong the patient's survival beyond that expected in the absence of such treatment. When applied to a single active ingredient (e.g., recombinant CD39) administered alone, the term refers to that ingredient alone. When applied to a combination, the term refers to the combined dose or amount of the active ingredients (whether administered serially or simultaneously in combination) that produces a therapeutic effect.

The phrase "dosing regimen" refers to the regimen used to treat a disease, for example, the dosing regimen used during the treatment of AKI (SA-AKI or CSA-AKI).

The phrase "means for administration" is used to refer to any device used to administer a drug systemically to a patient, including but not limited to prefilled syringes, vials and syringes, injection pens, autoinjectors, intravenous (i.v.) drip and injection bags, pumps, patch pumps, etc. Using these items, a patient can self-administer the drug (i.e., self-administer the drug) or a physician can administer the drug.

AKI will be determined based on the staging system outlined in the KDIGO acute kidney injury guidelines, for example, severe AKI. This classification system stages AKI into three levels based on the degree of sCr elevation and the degree and duration of oliguria; patients are categorized based on the worst outcome (sCr or oliguria). The following table categorizes the three stages of AKI: [ Table 1 ] Stages of AKI period of illness serum creatinine Urine volume 1 1.5-1.9 times baseline or an increase of ≥ 0.3 mg/dL (≥ 26.5 μmol/L) < 0.5 ml/kg/h for 6-12 hours 2 2.0-2.9 times baseline < 0.5 ml/kg/h for ≥ 12 hours 3 3.0 times baseline or serum creatinine increased to ≥ 4.0 mg/dL (≥ 353.6 μmol/L) or initiation of renal replacement therapy < 0.3 ml/kg/h for ≥ 24 hours or anuria for ≥ 12 hours

The diagnosis of sepsis can be made, for example, according to the criteria defined in the Third International Consensus Definition for Sepsis and Septic Shock (Sepsis-3), with the additional diagnosis of AKI (stage 1 or higher using KDIGO serum creatinine-based criteria).

The term "cardiac surgery" is used to refer to non-emergency open-chest major cardiopulmonary bypass (CPB) surgery with an expected CPB time of >1 hour, defined as any of the following: a. Combined coronary artery bypass grafting (CABG) surgery and surgery on one or more heart valves (Valve Surgery) b. Surgery on more than one heart valve (Valve Surgery) c. Surgery on the aortic root or ascending aorta, or in combination with an aortic valve d. Surgery on the aortic root or ascending aorta, combined with CABG and/or one or more valves.

Sequential Organ Failure Assessment Score: The SOFA (Sequential Organ Failure Assessment) score was developed to assess the acute onset of major illness at a population level and has been extensively validated as a tool for this purpose across a range of healthcare scenarios and settings. With the development of a new definition, the SOFA is now used as a key criterion for the diagnosis of sepsis syndrome at the individual patient level. This score is routinely calculated upon ICU admission and within each subsequent 24-hour period. The score consists of six criteria that reflect the function of specific organ systems (respiratory, cardiovascular, renal, neurologic, hepatic, and hematologic) and is assigned a score of 0-4.

Scores range from 0 to 24, with higher scores indicating greater functional impairment.

The SOFA score has been used in a range of clinical and research applications.

In the context of randomized controlled trials, a meta-analysis identified 25 studies that used the change in SOFA score from baseline or maximum to a specified time point and revealed a strong association between SOFA change and mortality—32% of the observed mortality effect was accounted for by the delta SOFA score. This association of SOFA scores at admission and during ICU stay with long-term outcomes led the EMA to accept the use of delta SOFA as a valid surrogate endpoint in exploratory clinical trials of participants with sepsis.

Acute Physiology and Chronic Health Evaluation II (APACHE-II) score is a system for classifying the severity of illness in patients. This score is administered within 24 hours of admission to the ICU or intermediate/high-density unit (HDU). The score, an integer between 0 and 71, is calculated based on several measurements: higher scores correspond to more severe illness and a higher risk of death in ICU patients.

Renal Replacement Therapy (RRT): RRT is recommended if at least one of the following criteria is met: 1. Anuria (little to no urine output for 6 hours) 2. Severe oliguria (urine output < 200 mL for more than 12 hours) 3. Hyperkalemia (potassium concentration > 6.5 mmol/L) 4. Severe metabolic acidosis (pH < 7.2 despite normal arterial carbon dioxide partial pressure or low PaCO2 partial pressure) 5. Volume overload 6. Significant azotemia (urea concentration > 30 mmol/L (> 84 mg/dL) or creatinine concentration > 300 µmol/L (> 3.4 mg/dL) 7. Clinical complications of uremia (e.g., encephalopathy, pericarditis, neuropathy)

"Major adverse renal events" (MAKE) are a composite endpoint that includes death, need for RRT, and worsening renal function (a decrease in eGFR of ≥ 25% from baseline). MAKEs can be assessed at designated time intervals/time points and are increasingly recognized as an important participant-centered outcome in studies conducted in severely ill participants. MAKE rates will be calculated at 30, 60, and 90 days. The number of participants with a composite endpoint of MAKE, defined as death, receipt of RRT, or a decrease in eGFR of ≥ 25% from baseline, will be included. eGFR will be calculated using the CKD-EPI formula, regardless of ethnicity.

In another preferred embodiment, the present disclosure relates to the use of recombinant CD39 according to the present invention for treating sepsis-associated acute renal injury (SA-AKI).

In another preferred embodiment, the present disclosure relates to the use of recombinant CD39 according to the present invention for treating cardiac surgery-associated acute kidney injury (CSA-AKI).

The following provides exemplary , non - limiting embodiments of the methods described in this disclosure.

Therapeutic proteins are typically formulated in aqueous form for administration or as a lyophilized form for reconstitution with a suitable diluent prior to administration. Proteins can be formulated as a lyophilized form or as an aqueous composition, such as in a pre-filled syringe.

Suitable formulations can provide aqueous pharmaceutical compositions or lyophilizates that can be reconstituted to deliver solutions with high concentrations of the therapeutic protein active ingredient and low levels of protein aggregation for delivery to patients. High protein concentrations are useful because they reduce the amount of material (dose) that must be delivered to the patient. The reduced dosage volume minimizes the time required to deliver a fixed dose to the patient. Aqueous compositions of the present invention having high protein concentrations are particularly suitable for subcutaneous administration.

Thus, the present invention provides aqueous pharmaceutical compositions suitable for administration in a subject (e.g., for subcutaneous administration), comprising a therapeutic protein.

When combined with a pharmaceutically acceptable carrier, the therapeutic protein can be used as a pharmaceutical composition. In addition to the therapeutic protein, such a composition may contain a carrier, various diluents, fillers, salts, buffers, stabilizers, cosolvents, and other materials known in the art. The characteristics of the carrier will depend on the route of administration. The pharmaceutical composition for use in the disclosed methods may also contain other therapeutic agents for treating the specific targeted disorder.

Because the indications treated according to the use described herein (e.g., treatment of SA-AKI) require immediate treatment, the treatment is not administered orally and may be given as an IV or liquid formulation.

Recombinant CD39 (hereinafter referred to as "CD39*") having the amino acid sequence of SEQ ID NO: 21 is an engineered, highly potent, soluble, and stable human recombinant CD39 enzyme. This enzyme hydrolyzes extracellular ATP and ADP into adenosine monophosphate (AMP), the rate-limiting step in the conversion of extracellular ATP to adenosine. AMP is further catalyzed into adenosine by the extracellular 5'-nucleotidase CD73. The enzymatic activities of CD39 and CD73 play a strategic role in calibrating the duration, amplitude, and chemical nature of purinergic signals delivered to immune cells, thereby driving the shift from an ATP-driven pro-inflammatory milieu to an adenosine-induced anti-inflammatory one. It is increasingly recognized that rebalancing this equilibrium may alter the course and outcome of several pathophysiological events, such as SA-AKI.

Exemplary methods for obtaining certain variants of CD39 (recombinant CD39, such as CD39*) are disclosed in WO 2020/016804. Example 2. Non-clinical study evaluation

CD39* has been extensively studied in preclinical models. In human whole blood, CD39* dose-dependently inhibits ATP/lipopolysaccharide (LPS)-induced IL-1β secretion and ADP-induced platelet aggregation. CD39* has also been shown to be beneficial in preclinical models of acute organ injury, including a mouse model of ischemia-reperfusion-induced AKI, where it dose-dependently protects renal function. This protection is manifested in renal architecture, renal inflammation, and acute tubular necrosis, hallmarks of human AKI pathophysiology (results not shown).

Furthermore, common biomarkers of kidney damage, function, and repair normalized to normal levels in kidney tissue and urine of animals experiencing AKI after CD39* treatment.

Target engagement biomarkers and proximal pharmacodynamics in the urine of animals after AKI showed a dose-dependent decrease in extracellular ATP and ADP, and a dose-dependent increase in extracellular AMP and adenosine.

The in vitro potency of CD39* in whole blood (WB) assays (LPS/ATP-induced IL-1β release and ADP-induced platelet aggregation) was demonstrated to be comparable in rats, minipigs, and humans. The similarity of pharmacology across species allows for the prediction and exploration of the expected safe exposure range based on animal safety and pharmacology data, and for achieving the expected therapeutic dose. The PD whole blood assays revealed an in vitro IC90 of approximately 1.66 to 2.66 μg/mL in humans. Example 3. Phase I , randomized, placebo-controlled, participant-blinded study in healthy volunteers

This Phase I, randomized, placebo-controlled, participant-blinded study was conducted in healthy volunteers in which a single IV dose of CD39* (0.1, 0.3, 1.0, and 2.0 mg/kg, as well as 4.0 mg/kg) was administered as a 2-hour infusion.

Administration of 0.1, 0.3, 1.0, and 2.0 mg/kg, as well as 4.0 mg/kg CD39* or placebo as a single IV infusion was well tolerated, and no safety issues related to the investigational medicinal product (IMP) were identified.

CD39* was safe and well tolerated, with no immunogenicity observed.

The PK results were fully consistent with the previously predicted CD39* serum concentrations.

Pathway engagement markers demonstrated dose-dependent and reversible inhibition of (i) ADP-induced platelet aggregation and (ii) LPS/ATP-induced IL-1β release in CD39*-treated subjects, with in vitro IC90s of 0.25 and 7.8 µg/mL for inhibition of platelet aggregation and IL-1β release, respectively.

CD39* induced a dose-dependent and reversible reduction in free inorganic pyrophosphate (PPi) levels at doses above 0.1 mg/kg, with maximal effects achieved within hours of administration.

Exposure determined by Cmax and AUCinf demonstrated dose-proportional behavior across this dose range. A 3-, 10-, and 20-fold increase in dose from 0.1 mg/kg to 0.3 mg/kg, 1.0 mg/kg, and 2.0 mg/kg resulted in a 3-, 12-, and 24-fold increase in mean Cmax and a 3-, 11-, and 17-fold increase in mean AUCinf, respectively.

Dose proportionality was also demonstrated by the model by evaluating the correlation between dose and Cmax and between dose and AUC.

The exposure of CD39* in human skin was studied across SAD cohorts 1 to 4 (0.1, 0.3, 1.0, and 2.0 mg/kg IV) by dosing CD39* in interstitial fluid collected by vesicular adsorption 24 and 168 hours after dosing. Interstitial fluid CD39* exposure increased in a fairly dose-proportional manner. Increasing the dose by 3-, 10-, and 20-fold increased mean interstitial fluid concentrations by 3-, 10-, and 33-fold, respectively, at 24 hours after dosing, and by 3-, 9-, and 18-fold, respectively, at 168 hours after dosing.

The results of the study showed that CD39* is safe and well tolerated. Example 4. A method for treating sepsis-related acute renal injury, the method comprising administering a therapeutically effective amount of recombinant CD39* to a subject in need of such treatment

This multicenter, participant- and investigator-blinded, randomized, placebo-controlled study aims to characterize the PK/PD profile and evaluate the safety and efficacy of the human recombinant CD39 enzyme CD39* in hospitalized adult participants diagnosed with sepsis and acute renal injury (AKI). Approximately 20 participants will be randomly assigned to the study. The study consists of a screening period (up to 48 hours), a treatment period (Day 1), and a post-treatment period (Days 2 to 90).

Screening will be conducted during hospitalization in the intensive care unit (ICU) or moderate/high dependency unit (HDU), where potential participants will be screened for the presence of sepsis and AKI based on the Sequential Organ Failure Assessment (SOFA) and Kidney Disease Improving Global Outcomes (KDIGO) scores.

Participants will be randomly assigned to receive either 2 mg/kg of CD39* or placebo using a 3:1 (CD39*:placebo) allocation ratio. A single dose of study treatment will be administered via a 2-hour IV infusion at the randomization visit, blinded to the participant and investigator. Study treatment must be administered within 48 hours of the first diagnosis of SA-AKI, but preferably as soon as possible. Two PK samples will be collected on Day 1: one before study treatment and one immediately (within 15 minutes) after completion of the IV infusion. Additional samples will be collected on Days 2, 3, 5, 8, 14, 30, 60, and 90, ideally at the same time as the start of CD39* dosing on Day 1. All participants will be followed up for a total of 90 days.

Safety will be monitored throughout the study. All adverse events (AEs) and serious adverse events (SAEs) will be collected and reported according to standard procedures and local regulations.

At 2 mg/kg, the expected systemic exposure and interstitial exposure in relevant tissues are thought to inhibit purinergic signaling and provide beneficial pharmacological effects. In a two-week minipig study, the total exposure (AUC inf ) in humans following a single dose of 2 mg/kg was approximately 10.4-fold lower than the total exposure at the NOAEL of CD39* after four IV doses of 10 _mg /kg every 4 days (a 2.6-fold margin compared to the AUC _tau,ss after the fourth dose in minipigs at steady-state), and C _max showed a 7-fold margin above the NOAEL. Participants will be followed for a total of 90 days.

Unless otherwise specified at the time of randomization (Day 1), eligible participants must meet all of the following criteria at the screening assessment: 1. Must provide signed informed consent prior to study participation. 2. Age ≥ 18 years and ≤ 85 years. 3. Admitted to the ICU or moderate/HDU. 4. Diagnosis of sepsis according to the criteria defined by the third international consensus definition for sepsis and septic shock (SESEP-3) based on: • suspected or confirmed infection • SOFA score of 2 or more (excluding the renal component) 5. Diagnosis of AKI stage 1 or higher at the time of randomization according to the following criteria: • absolute increase in serum or plasma creatinine (CR) of ≥ 0.3 mg/dL (≥ 26.5 μmol/L) within 48 hours or presumed to have occurred within the past 48 hours compared to the reference creatinine baseline For hospital-acquired AKI, a stable serum creatinine obtained in the hospital before the onset of AKI should be used as the reference baseline. Otherwise, the baseline serum creatinine should be prioritized in the following order: 1. Median value within 3 months of admission. If unavailable: 2. Median value between 3 and 6 months before admission. If unavailable: 3. On admission.

Unless otherwise specified at randomization (Day 1), participants who met any of the following criteria at the screening assessment were ineligible for enrollment in the study. 1. Not expected to survive 24 hours. 2. Not expected to survive 30 days due to a medical condition other than SA-AKI. 3. Documented history of CKD with an estimated GFR < 45 ml/min before the development of AKI. 4. Currently receiving RRT or a decision to start RRT within 24 hours of admission. 5. Weight less than 40 kg or greater than 125 kg. 6. With life support limitations (e.g., not capable of CPR, not capable of dialysis, not capable of intubation). 7. AKI, as defined by the AKI inclusion criteria, persisting for more than 48 hours before study drug administration. 8. AKI, as indicated by clinical manifestations (e.g., prolonged oliguria or severe renal failure (e.g., serum creatinine > 4 mg/dL) on admission and no history of CKD), persisting for more than 48 hours before study drug administration. 9. Evidence of recovery from AKI, as determined by the investigator's clinical judgment, prior to randomization. 10. AKI is most likely due to causes other than sepsis, such as nephrotoxic drugs (nonsteroidal anti-inflammatory drugs (NSAIDs), contrast agents, aminoglycosides), other medical conditions (e.g., heart failure, liver failure, acute abdominal aortic aneurysm, dissection, renal artery stenosis), or urinary tract obstruction. 11. A history of documented (biopsy-confirmed) or suspected acute or subacute renal disease, such as rapidly progressive glomerulonephritis (RPGN) and acute interstitial nephritis (AIN). 12. Patients undergoing nephrectomy. 13. Patients receiving dual antiplatelet therapy. 14. Patients with thrombocytopenia (platelet count < 100,000 μL) at screening or otherwise at high bleeding risk as determined by the investigator. 15. Immunosuppressed Patients: • History of immunodeficiency or known HIV positive test. • Patients currently receiving immunosuppressive agents or chronic high-dose (high-dose therapy for more than 2 weeks) steroids equivalent to prednisone/prehydrocortisone 0.5 mg/kg/day, including solid organ transplant recipients. This may include patients in septic shock treated with hydrocortisone (e.g., 3 × 100 mg). 16. Active hepatitis (defined as (a) abnormal liver enzymes (alanine aminotransferase (ALT), gamma-glutamyl transferase (GGT), ALP > 3 times the upper limit of normal (ULN)) or (b) positive hepatitis B virus (HBV) or hepatitis C virus (HCV) serology for active hepatitis B or C infection), or patients with advanced chronic liver disease confirmed by a Child-Pugh score of 10-15 (grade C). 17. Acute pancreatitis of undetermined infectious origin. 18. Active hematologic malignancy (previously untreated hematologic malignancy is permitted). 19. Burns requiring ICU treatment. 20. Septicemia attributable to confirmed COVID-19. 21. Use of other investigational drugs within 5 half-lives (30 days (e.g., small molecules)) at the time of enrollment and/or the expected pharmacodynamic effect has not recovered to baseline (e.g., biologics), whichever is longer; or a longer period of use of other investigational drugs if required by local regulations. 22. History of hypersensitivity reaction to the study treatment or its formulations, or to drugs of similar chemical classes. 23. Any medical condition that, in the judgment of the investigator, could significantly increase the safety risk to the participant by participating in this study. 24. Women with a positive pregnancy test, who are pregnant, or who are breastfeeding. 25. Women of childbearing potential are defined as all women who are physiologically capable of becoming pregnant, unless they are using highly effective contraception while on study treatment and for 38 days after stopping study treatment. Highly effective contraceptive methods include: •   Total abstinence (when consistent with the participant's preferred and daily lifestyle). Cyclic abstinence (e.g., calendar, ovulation-based, temperature-based, post-ovulation methods) and ejaculation are not acceptable methods of contraception. •   Female sterility (bilateral oophorectomy, with or without hysterectomy), total hysterectomy, or bilateral tubal ligation at least six weeks before starting the study drug. In the case of oophorectomy alone, only if the female reproductive status has been confirmed by subsequent hormonal assessment. •   Male sterility (at least 6 months before screening). For female participants in the study, the vasectomized male partner should be the participant's only partner. •   Use of oral (estrogen and progesterone), injectable, or implantable hormonal contraceptive methods or placement of an intrauterine device (IUD) or intrauterine system (IUS), or other hormonal contraceptive methods with similar efficacy (failure rate < 1%), such as hormonal vaginal rings or transdermal hormonal contraception.

Study treatment will be administered by investigators at the clinical site as a 2-hour IV infusion (approximately) via an infusion syringe according to designated study procedures. Example 5. A randomized, multicenter, placebo-controlled, participant- and investigator-blinded study to evaluate the safety, tolerability, and efficacy of a single IV infusion of CD39* in adult patients at risk for acute renal injury after cardiac surgery

This is a non-validation, randomized, multicenter, placebo-controlled study with blinded participants and investigators. Approximately 120 adult patients at risk for acute renal injury after cardiac surgery will be enrolled in the study.

There will be two groups in the study: • Placebo • CD39* 4 mg/kg

Participants entering this study will be managed according to current medical and surgical standards of care and in accordance with standard institutional procedures. The study will consist of a preoperative period (screening visit), a treatment period (Day 1), and a follow-up period (Day 2 to Day 90). Participants will be followed up on Days 2, 3, 4, 5, 6, and 8 in the hospital (or at home/rehabilitation center until discharged earlier on Day 8) and then as outpatients until the end of the study (Day 30 and Day 90 visits).

On Day 1, participants will undergo cardiac surgery. Randomization will occur once cardiopulmonary bypass time is confirmed to be equal to or greater than 60 minutes. Participants will be stratified according to eGFR >= 30 mL/min/1.73 m² versus eGFR < 30 mL/min/1.73 m². Once wound closure is achieved and the absence of ongoing bleeding is confirmed, participants will receive a single dose of CD39* or placebo. Administration should begin as soon as possible after wound closure and no later than 60 minutes after wound closure.

In this study, patients will receive either a 4 mg/kg dose of study drug or placebo via a 2-hour intravenous infusion in a double-blind fashion. The 4 mg/kg dose represents the highest safe dose tested in healthy volunteers, and the expected systemic and interstitial exposures are thought to inhibit purinergic signaling and provide beneficial pharmacological effects. In a 2-week minipig study, a single dose of 4 mg/kg resulted in a total human exposure (AUCinf) approximately 5.2-fold lower than the total exposure at the NOAEL of CD39* after four i.v. doses of 10 mg/kg every 4 days (a 1.3-fold margin compared to the AUC after the fourth minipig dose), and Cmax demonstrated a 3.5-fold margin above the NOAEL. Patients will be followed for a total duration of 90 days.

Participants eligible for inclusion in this study must meet all of the following criteria: 1. Must provide signed informed consent prior to study participation. 2. Participants must be able to communicate well with the investigator and understand and comply with study requirements. 3. Male and female patients must be ≥ 45 years of age at screening. 4. Participants must weigh a minimum of 50 kg and a maximum of 150 kg to participate in the study and have a body mass index (BMI) less than 40. BMI = Weight (kg) / [Height (m)] ² 5. Vital signs (systolic and diastolic blood pressure and pulse rate) will be assessed in the sitting position at screening. Seated vital signs should be within the following ranges: a. Oral temperature between 35.0°C and 37.5°C b. Blood pressure (systolic 100-160 mmHg, diastolic <100 mmHg) c. Pulse rate (50-100/min) stable, regardless of medication, as assessed by the Investigator. If vital signs are outside these ranges, the Investigator may obtain two additional readings, allowing for up to three consecutive assessments. In order for a participant to qualify, at least the final reading must be within the ranges provided above. 6. No known increase in SCr of ≥ 25% at the Screening Visit compared to the previous value recorded no more than 6 weeks earlier in the on-site local laboratory using standard testing methods. 7. eGFR < 60 mL/min/1.73 m2 (CKD stage 3A or higher) at screening (calculated using the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI 2021) formula). 8. Non-emergency open-chest major cardiopulmonary bypass (CPB) surgery with an expected CPB time > 1 hour, defined as any of the following: a. Combined coronary artery bypass grafting (CABG) surgery and surgery on one or more heart valves (Valve Surgery). b. Surgery on more than one heart valve (Valve Surgery). c. Surgery on the aortic root or ascending portion of the aorta, or in combination with an aortic valve. d. Surgery involving the aortic root or ascending portion of the aorta combined with CABG and/or one or more valves. Note 1: If CABG or single-valve surgery is performed alone, participants are required to have at least one additional AKI risk factor: i. Medication-dependent diabetes mellitus, defined as type 1 or type 2, prior to hospitalization for cardiac surgery. ii. Proteinuria, defined as a urine albumin-to-creatinine ratio > 300 mg/g (30 mg/mmol) or urine albumin excretion > 300 mg/24 hours at screening. iii. Age ≥ 70 years at screening. iv. History of congestive heart failure requiring hospitalization. NOTE 2: If minimally invasive cardiac surgery (MICS) performed with a small thoracotomy meets the definition of open chest intervention, patients undergoing MICS may be enrolled if the CPB time is > 1 hour and they meet inclusion criterion 8. NOTE 3: Transcatheter aortic valve implantation (TAVI) or transcatheter aortic valve replacement (TAVR) is permitted only when performed in combination with a procedure defined in inclusion criterion 8.

Participants who meet any of the following criteria are not eligible for inclusion in this study. 1. eGFR < 15 mL/min/1.73 m2 at screening (calculated using the CKD-EPI 2021 formula). 2. Currently receiving renal replacement therapy. 3. Patients at risk for bleeding at screening, defined as: - A history of bleeding with suspected or confirmed bleeding disorders, or any other high bleeding risk determined by the investigator - Thrombocytopenia: Platelet count < 100 x 10 ⁹ /L - Thrombopenia: ADP-induced platelet aggregation less than 60% - Preexisting coagulation factor deficiency: including but not limited to fibrinogen < 2.5-2.8 g/L 4. Any emergency surgery performed less than 30 days prior to screening, including aortic stenosis and/or severe congenital heart defect. 5. Planned cardiac surgery with weaning of CPB or hypothermic circulatory arrest. 6. Planning to undergo TAVI or TAVR only, or single-vessel, minimally invasive direct coronary artery bypass grafting (MIDCAB) off-pump surgery, or left ventricular assist device (LVAD) implantation (Note: See Inclusion Criteria 8). 7. Cardiogenic shock or hemodynamic instability within 4 weeks prior to surgery, requiring inotropes or vasopressors, or a mechanical device such as an intra-aortic balloon pump (IABP). 8. Requirement of any of the following within 4 weeks prior to cardiac surgery: defibrillator, mechanical ventilation, IABP, LVAD, or other form of mechanical circulatory support (MCS). 9. Received cardiopulmonary resuscitation (CPR) within 30 days prior to cardiac surgery. 10. Recent stent implantation ≤ 30 days prior to cardiac surgery, or other conditions requiring reinitiation of P2Y12 inhibitor therapy within 48 hours or less after surgery. 11. Cardiopulmonary bypass (CPB) duration < 60 minutes. 12. Patients with active bleeding at the end of surgery, as assessed by the surgical team. 13. Use of other study drugs at the time of enrollment, or within 5 half-lives of other study drugs at the time of enrollment, or the expected PD effect has not recovered to baseline, whichever is longer; or if local regulations require the use of other study drugs for a longer period of time. 14. History of hypersensitivity reaction to any study treatment or formulation, or to drugs of similar chemical class. 15. Known history (within 6 months prior to screening) or current clinically significant arrhythmias associated with dizziness, dyspnea, or hemodynamic instability. 16. Patients undergoing nephrectomy. 17. Concomitant use of medications known to prolong the QT interval, unless such medications can be permanently discontinued during the study (QT > 500 ms at the discretion of the investigator). 18. Taking medications prohibited by the protocol. 19. History of malignancy of any organ system (excluding localized basal cell carcinoma of the skin or in situ cervical carcinoma) in the past 5 years, treated or untreated (regardless of whether there is evidence of local recurrence or metastasis). 20. Donation or loss of > 450 mL of blood or > 200 mL of plasma within four weeks or longer prior to drug administration, if required by local regulations. 21. Any history of organ or cell transplantation requiring active immunosuppressive therapy that may interfere with renal function (e.g., calcineurin inhibitors). 22. Persistent sepsis or a history of sepsis within the past 8 weeks, or an untreated confirmed infection prior to the screening visit. Sepsis is defined as the presence of a documented pathogen, accompanied by fever or hypothermia and hypoperfusion or hypotension. 23. Stage IV heart failure as defined by the New York Heart Association (NYHA) Functional Classification. 24. Left ventricular ejection fraction < 30% based on the most recent available echocardiogram. 25. Recent history (within the past three years) and/or recurrence of autonomic dysfunction (e.g., recurrence of syncope, palpitations, etc.). 26. Any surgical or medical condition that could significantly alter the absorption, distribution, metabolism, or excretion of the drug, or that could endanger the participant during study participation. The investigator should make this decision based on the participant's medical history and/or clinical or laboratory evidence of any of the following: • Inflammatory bowel disease, peptic ulcers, gastrointestinal disease, including rectal bleeding; • Major gastrointestinal surgery, such as gastrectomy, gastroenterostomy, or bowel resection; • Pancreatic injury or pancreatitis; • Abnormal liver function tests indicating liver disease or damage: ALT (SGPT), AST (SGOT), GGT, alkaline phosphatase, and serum bilirubin will be measured. • Evidence of urinary tract obstruction or dysuria at screening. If necessary, laboratory tests may be repeated once before randomization (as soon as possible) to rule out any laboratory errors. 27. History of immunodeficiency. (Assessed by local testing?) Also includes tuberculosis? 28. Chronic infection with hepatitis B (HBV) or hepatitis C (HCV) (history). A positive HBV surface antigen (HBsAg) test, or, if following standard local practices, a positive HBV core antibody test, excludes participants. Active hepatitis defined by active AST/ALT? (From sepsis testing?) 29. History of drug abuse or unhealthy alcohol consumption within 12 months prior to drug administration ^# or evidence of such abuse by laboratory testing performed during screening. ^#Unhealthy drinking is defined as a history of alcohol misuse/abuse or current alcohol misuse/abuse, defined as "five or more drinks on the same occasion on each of five or more days in the past 30 days." 30. Pregnant or lactating (lactating) women. 31. Women of childbearing potential, defined as all women who are physiologically capable of pregnancy, unless they use highly effective contraception while on study treatment and until the end of the study. Highly effective contraceptive methods include: • Complete abstinence (when this is consistent with the participant's preferred and daily lifestyle). Cyclic abstinence (e.g., calendar, ovulation-based, temperature-based, post-ovulation methods) and ejaculation outside the body are not acceptable methods of contraception. • Bilateral tubal ligation, female sterilization (with bilateral oophorectomy, with or without hysterectomy), or total hysterectomy at least six weeks before study treatment. In the case of oophorectomy alone, only if the female reproductive status has been confirmed by subsequent hormonal level assessment. • Male sterilization (at least 6 months before screening). For female participants in the study, the vasectomized male partner should be the participant's only partner. • Use of oral (estrogen and progesterone), injectable, or implantable hormonal contraceptive methods or placement of an intrauterine device (IUD) or intrauterine system (IUS) or other hormonal contraceptive methods with similar efficacy (failure rate < 1%), such as hormonal vaginal rings or transdermal hormonal contraception. Example 6: A multicenter, randomized, double-blind, placebo-controlled, four-arm, parallel-group, Phase 2b dose-finding study to investigate the safety and efficacy of CD39* in the treatment of patients with sepsis-associated acute kidney injury ( SA-AKI )

This is a double-blind, placebo-controlled, four-arm, parallel-group, Phase 2b dose-finding study. Approximately 320 participants will be randomly assigned in a 3:1:1:3 ratio to one of three single CD39* doses (4 mg/kg, 2 mg/kg, 1 mg/kg) or placebo. Key inclusion criteria • Age ≥ 18 years to ≤ 85 years • Admission to the ICU or intermediate care unit/high dependency unit (HDU) • Criteria defined according to the third edition of the International Consensus Definitions for Sepsis and Septic Shock (Sepsis-3), based on a diagnosis of sepsis: - suspected or confirmed infection, and - an acute increase in the SOFA score of 2 points or more (excluding the renal component). The baseline SOFA score should be considered zero unless the participant is known to have pre-existing organ dysfunction (acute or chronic) before the infection. • AKI stage 1 or higher diagnosed at randomization according to the following criteria: an absolute increase in serum or plasma creatinine ≥ 0.3 mg/dL (≥ 26.5 μmol/L) compared with the reference pre-septic creatinine within 48 hours or presumed to have occurred within the past 48 hours. • For hospital-acquired AKI, a stable serum creatinine obtained in the hospital before the diagnosis of AKI should be used as the reference serum creatinine. • For patients from the community, the reference pre-septic serum creatinine should be estimated using the following priority: 1. Most recent value within 3 months of admission. If unavailable: 2. Most recent value between 3 and 12 months before admission. If unavailable: 3. Key exclusion criteria at admission • Documented history of chronic kidney disease (CKD) with eGFR < 45 mL/min before admission. • eGFR < 45 mL/min on admission and no other reference serum eGFR in the last 12 months • Currently receiving renal replacement therapy (RRT) or a decision to start RRT within 24 hours of admission • Presence of sepsis as determined by sepsis inclusion criteria for more than 72 hours before ICU admission • AKI as determined by AKI inclusion criteria within 48 hours of ICU admission • Inability to administer study drug within 24 hours of AKI as determined by AKI inclusion criteria • Presence of AKI as determined by the investigator, as clinically indicated (e.g., prolonged oliguria or severe renal dysfunction on admission and no history of CKD), for more than 24 hours before study drug administration Prior to randomization, participants had evidence of recovery from AKI based on the investigator's clinical judgment: • A history of documented (biopsy-confirmed) or suspected acute or subacute renal disease, such as rapidly progressive glomerulonephritis (RPGN) and acute interstitial nephritis (AIN); • Participants who had thrombocytopenia (platelet count < 50,000 μL) at screening or were otherwise at high bleeding risk as determined by the investigator. One or more primary objectives: • To assess the dose-response relationship of CD39* to renal function as measured by creatinine clearance (CrCl) in subjects with SA-AKI. One or more secondary objectives: • To assess the effect of CD39* compared to placebo in reducing major renal adverse events • To assess the effect of CD39* compared to placebo in improving renal function • To assess the effect of CD39* compared to placebo in improving global health status • To assess the safety and tolerability of CD39* compared to placebo Secondary estimates are defined by assessing the effect of study treatment on the following endpoints and summary measures: • MAKE (death, RRT, ≥ 25% reduction in eGFR) at Day 90: the summary measure is the odds ratio for participants who have MAKE. • Weighted mean of the area under the curve of time-adjusted endogenous serum creatinine from Day 1 to Day 14 and from Day 1 to Day 30: the summary measure is the geometric mean. • Weighted mean of the time-adjusted area under the curve for endogenous serum cystin-C from Days 1 to 14 and from Days 1 to 30: The summary measure is the geometric mean. • Weighted mean of the time-adjusted area under the curve for endogenous creatinine clearance from Days 5 to 14 (AUC5-14). The estimation framework was the same as for the primary endpoint. • Any use of RRT during the study: The summary measure is the odds ratio for subjects who received RRT. • RRT dependence at Day 90: The summary measure is the odds ratio for subjects who used RRT. • Number of days alive and RRT-free during the study: The summary measure is the mean number of days alive and RRT-free. • Proportion of subjects with a ≥ 25% decrease in eGFR at Day 90: The summary measure is the odds ratio for subjects with a ≥ 25% decrease in eGFR. • KDIGO AKI duration: The summary measure is the change from baseline in KDIGO AKI duration on Day 14. • SOFA score change from baseline to Day 30: The summary measure is the mean change from baseline in SOFA score.

Potential study participants will have a recent diagnosis of sepsis and a subsequent diagnosis of AKI according to criteria defined by the third edition of the International Consensus Definitions for Sepsis and Septic Shock ( SEPTEMIA -3). SEQ ID NO describe sequence 1 Human recombinant CD39 variants TQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAREVIPRSQHQETPVYLGATAGMRLLRMESEE LADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQAL WQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVMKF LNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEYCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 2 Human recombinant CD39 variants TQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAMEVIPRSQHQETPVYLGATAGMRLLRMESEE LADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQAL WQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVMKF LNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEYCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 3 Human recombinant CD39 variants TQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAREVIPRSQHQETPVYLGATAGMRLLRMESEE LADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQAL WQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYSVMKF LNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEYCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 4 Human recombinant CD39 variants TQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAMEVIPRSQHQETPVYLGATAGMRLLRMESEE LADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQAL WQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVMKF LNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVNEKYLSEFCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 5 Human recombinant CD39 variants TQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAMEVIPRSQHQETPVYLGATAGMRLLRMESEE LADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQAL WQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVMKF LNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEFCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 6 Human recombinant CD39 variants TQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAREVIPRSQHQETPVYLGATAGMRLLRMESEE LADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQAL WQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVMKF LNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEFCFSGTYILSLLQQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 7 Human recombinant CD39 variants TQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAMEVIPRSQHQETPVYLGATAGMRLLRMESEE LADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQAL WQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVMKF LNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEFCFSGTYILSLLQQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 8 Human recombinant CD39 variants TQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAMEVIPRSQHQETPVYLGATAGMRLLRMESEE LADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQAL WQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYSVMKF LNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEYCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 9 Human recombinant CD39 variants APTSSSTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAMEVIPRSQHQETPVYLGATAGMRLLRM ESEELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKD QALWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVM KFLNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEFCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 10 Human recombinant CD39 variants APTSSSTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAREVIPRSQHQETPVYLGATAGMRLLRM ESEELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKD QALWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVM KFLNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEFCFSGTYILSLLQQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 11 Human recombinant CD39 variants APTSSSTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAMEVIPRSQHQETPVYLGATAGMRLLRM ESEELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKD QALWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVM KFLNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEFCFSGTYILSLLQQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 12 Human recombinant CD39 variants APTTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAREVIPRSQHQETPVYLGATAGMRLLRMES EELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQA LWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVMK FLNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEFCFSGTYILSLLQQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 13 Human recombinant CD39 variants APTTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAMEVIPRSQHQETPVYLGATAGMRLLRMES EELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQA LWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVMK FLNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEFCFSGTYILSLLQQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 14 Human recombinant CD39 variants APTTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAMEVIPRSQHQETPVYLGATAGMRLLRMES EELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQA LWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVMK FLNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEYCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 15 Human recombinant CD39 variants APTTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAMEVIPRSQHQETPVYLGATAGMRLLRMES EELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQA LWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYSVMK FLNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEYCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 16 Human recombinant CD39 variants APTSSSTKKTQLTSSGTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAREVIPRSQHQETPVYLGAT AGMRLLRMESEELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFL CYGKDQALWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYF VMKFLNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEYCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 17 Human recombinant CD39 variants APTSSSTKKTQLTSSTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAMEVIPRSQHQETPVYLGATA GMRLLRMESEELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLC YGKDQALWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYF VMKFLNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVNEKYLSEFCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 18 Human recombinant CD39 variants APTSSSTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAMEVIPRSQHQETPVYLGATAGMRLLRM ESEELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKD QALWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYSVM KFLNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEYCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 19 Human recombinant CD39 variants APTTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAMEVIPRSQHQETPVYLGATAGMRLLRMES EELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQA LWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVMK FLNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEFCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST 20 Human recombinant CD39 variants APTTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAREVIPRSQHQETPVYLGATAGMRLLRMES EELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQA LWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVMK FLNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEYCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST twenty one Human recombinant CD39 variants APTTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAREVIPRSQHQETPVYLGATAGMRLLRMES EELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLCYGKDQA LWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYSVMK FLNLTSEKVSQEKVTEMMKKFCAQPWEEIKTSYAGVKEKYLSEYCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHST twenty two Human recombinant CD39 MEDTKESNVKTFCSKNILAILGFSSIIAVIALLAVGLTQNKALPENVKYGIVLDAGSSHTSLYIYKWPAEKENDTGVVHQVEECRVKGPGISKFVQKVNEIGIYLTDCMERAREVIPRSQHQETPVY LGATAGMRLLRMESEELADRVLDVVERSLSNYPFDFQGARIITGQEEGAYGWITINYLLGKFSQKTRWFSIVPYETNNQETFGALDLGGASTQVTFVPQNQTIESPDNALQFRLYGKDYNVYTHSFLC YGKDQALWQKLAKDIQVASNEILRDPCFHPGYKKVVNVSDLYKTPCTKRFEMTLPFQQFEIQGIGNYQQCHQSILELFNTSYCPYSQCAFNGIFLPPLQGDFGAFSAFYFVMKFLNLTSEKVSQEKV TEMMKKFCAQPWEEIKTSYAGVKEKYLSEYCFSGTYILSLLLQGYHFTADSWEHIHFIGKIQGSDAGWTLGYMLNLTNMIPAEQPLSTPLSHSTYVFLMVLFSLVLFTVAIIGLLIFHKPSYFWKDMV

without

without

without

TW202404627A_112128207_SEQL.xml

Claims (19)

A use of recombinant CD39 for preparing a medicament for treating or preventing acute kidney injury (AKI) in a subject in need thereof, wherein the recombinant CD39 has the amino acid sequence of SEQ ID NO: 21. The use as described in claim 1, wherein the AKI is severe AKI. The use of claim 1, wherein at least one symptom of AKI in the individual is reduced or eliminated. The use of claim 1, wherein the severity of AKI is reduced by at least one stage as defined by the modified AKI Network (AKIN) criteria for serum creatinine. The use according to any one of claims 1 to 4, wherein the recombinant CD39 is administered at a dose of 0.1 mg/kg to 10 mg/kg of the recombinant CD39. The use as described in claim 5, wherein the recombinant CD39 is administered at a dose of 4 mg/kg of the recombinant CD39. The use as described in claim 5, wherein the recombinant CD39 is administered at a dose of 5 mg/kg of the recombinant CD39. The use according to any one of claims 1 to 4, wherein the recombinant CD39 is administered intravenously (IV). The use of any one of claims 1 to 4, wherein the recombinant CD39 is administered within 48 hours after the individual is first diagnosed with AKI. The use of any one of claims 1 to 4, wherein the recombinant CD39 is administered within 36 hours after the individual is first diagnosed with AKI. The use of any one of claims 1 to 4, wherein the recombinant CD39 is administered within 24 hours after the individual is first diagnosed with AKI. The use of any one of claims 1 to 4, wherein the recombinant CD39 is administered within 12 hours after the individual is first diagnosed with AKI. The use according to any one of claims 1 to 4, wherein the AKI is sepsis-associated acute renal injury (SA-AKI). The use according to any one of claims 1 to 4, wherein the AKI is cardiac surgery-associated AKI (CSA-AKI). The use according to any one of claims 1 to 4, wherein the recombinant CD39 is used to prevent or treat one or more symptoms associated with AKI. The use according to any one of claims 1 to 4, wherein the recombinant CD39 is administered at a dose of 1 mg/kg, 2 mg/kg, 3 mg/kg, 4 mg/kg or 5 mg/kg. The use of any one of claims 1 to 4, wherein the recombinant CD39 is administered to the individual as a single therapy. The use according to any one of claims 1 to 4, wherein the recombinant CD39 is co-administered with another therapeutic agent. The use of claim 18, wherein the recombinant CD39 is co-administered with another therapeutic agent selected from steroids, metalloproteinase inhibitors, serine protease inhibitors, local anesthetic emulsions, proliferation factor inhibitors, antinausea drugs or antibiotics.