combination

Abstract

The present invention provides a combination comprising (a) a sulfonylurea and (b) at least one of the following components: (i) an insulin modulator and (ii) an aldosterone antagonist, such as a combination comprising (a) glibenclamide, or a structural or functional analog thereof, and (b) at least one of the following components: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) potassium canrenoate, or a structural or functional analog thereof. The combination is suitable for treating stroke and other neurodegenerative disorders, and for treating and / or preventing ischemia and / or reperfusion injury in various vital organs, including the brain and heart. Further aspects of the present invention relate to pharmaceutical products and compositions comprising the combination according to the present invention, as well as methods of treatment using the same.

Description

[Technical Field]

[0001] The present invention provides combinations suitable for the treatment of stroke and neurodegenerative disorders, as well as for treating and / or preventing ischemia and / or reperfusion injury in various vital organs, including the brain and heart.

[0002] Further aspects of the invention relate to pharmaceutical products and compositions containing said combinations, and to methods of treatment using same. [Background technology]

[0003] Strokes are caused by a lack of blood flow in the brain (ischemic stroke) or bleeding in the brain (hemorrhagic stroke), both of which result in brain cell death. According to the World Health Organization, stroke is the second leading cause of death worldwide, accounting for approximately 6 million deaths in 2016. According to the Global Burden of Disease study (Johnson CO et al. Lancet Neurol. 2019;18:439-58), there were 13.7 million new stroke cases and 80.1 million prevalent stroke cases worldwide in 2016. The high burden of stroke worldwide suggests that primary prevention strategies are not widely implemented or are not sufficiently effective. Guidelines for the management of acute ischemic stroke are available (Powers WJ, et al. Stroke. 2018;49:e46-e99). Interestingly, the guidelines conclude that currently, no pharmacological or non-pharmacological treatments with putative neuroprotective properties have demonstrated efficacy in improving outcomes after ischemic stroke, and therefore, other neuroprotective agents are not recommended. Guidelines for the management of hemorrhagic stroke also exist, but these guidelines do not recommend any therapy for managing the neurodegenerative consequences of hemorrhagic stroke other than rehabilitation (Hemphill JC 3rd, et al. Stroke. 2015; 46: 2032-2060).

[0004] The above data clearly demonstrate the current lack of effective pharmacological treatments for ischemic or hemorrhagic stroke and the need for neuroprotective treatments for stroke patients. Effective treatment of stroke-related reperfusion injury may provide neuroprotection. However, to date, attempts to develop effective neuroprotective treatments for stroke patients based on reducing reperfusion injury have been unsuccessful (Savitz SI, et al. Stroke. 2017; 48: 3413-3419, Patel RAG, et al. Prog Cardiovasc Dis. 2017; 59: 542-548). Previous research in the field of cardioprotection and reperfusion injury has revealed the surprising discovery that combination therapies not indicated for the treatment of cardiovascular disorders can provide significant synergistic effects in protecting against myocardial reperfusion injury when used at lower dose levels than those indicated for these other conditions (WO 2017 / 077378; U.S. Pat. No. 10,172,914; Genesis Pharma SA).

[0005] Neuroprotective therapies are expected to be useful in the treatment of neurodegenerative disorders. Neurodegenerative disorders are characterized by the progressive loss of neuronal structure or function, ultimately leading to neuronal death. Neurodegenerative disorders include currently incurable diseases such as Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, and vascular dementia. Alzheimer's disease showed the highest increase (46.2%) among neurological disorders worldwide between 2007 and 2017, resulting in 2.51 million deaths in 2017 (Roth GA, et al. Lancet. 2018; 392:1736-88). In the same study, Parkinson's disease also showed a 38.3% increase, resulting in 340,600 deaths worldwide in 2017. The prevalence of amyotrophic lateral sclerosis is 5.40 per 100,000 in Europe, 3.40 in the United States, and 2.34 in Asia (Marin B, et al. Int J Epidemiol 2017;46:57-74). Huntington's disease is a genetic neurological disorder considered rare, with prevalence ranging from 0.4 per 100,000 in Asia to 7.3 per 100,000 in North America (Rawlins MD. Neuroepidemiology. 2016;46:144-53). Vascular dementia is dementia caused by problems with blood supply to the brain, typically a series of minor strokes, leading to progressively worsening cognitive decline. This term refers to a syndrome consisting of a complex interaction of cerebrovascular disease and risk factors, leading to structural changes in the brain due to strokes and lesions, and consequent changes in cognition. Vascular dementia is the second most common dementia in the elderly after Alzheimer's disease (AD) (Battistin L, (December 2010). Neurochemical Research. 35 (12): 1933-8.; "Vascular Dementia: A Resource List"). The prevalence of the disease is 1.5% in Western countries and approximately 2.2% in Japan. This accounts for 50% of all dementia cases in Japan, 20%-40% in Europe, and 15% in Latin America.Twenty-five percent of stroke patients develop new-onset dementia within one year of the stroke. One study found that the prevalence of vascular dementia in the United States is 2.43% among all people over the age of 71, while another study found that the prevalence of dementia doubles every 5.1 years (Plassman BL, (2007). Neuroepidemiology. 29 (1-2); Jorm AF (November 1987), Acta Psychiatrica Scandinavica. 76 (5): 465-79.). Currently, there are no medications specifically approved for the prevention or treatment of vascular dementia. Currently approved therapies for Alzheimer's disease provide limited benefit (Atri A. Med Clin North Am. 2019; 103: 263-293). Although multiple pharmacological treatments are available to manage the motor and non-motor symptoms of Parkinson's disease, they are essentially symptomatic and ultimately induce dyskinesia, while none of these treatments offer neuroprotection (Chaudhuri KR, et al. Parkinsonism Relat Disord. 2016; 33 (Suppl 1): S2-S8). Currently, there are only two approved drugs (riluzole and edaravone) that modestly slow the progression of amyotrophic lateral sclerosis, and no approved therapies for Huntington's disease. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] International Publication No. 2017 / 077378 Brochure [Patent Document 2] U.S. Patent No. 10,172,914 [Non-patent literature]

[0007] [Non-Patent Document 1] Johnson CO et al. Global, regional, and national burden of stroke, 1990-2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet Neurol. 2019; 18 : 439-458 [Non-Patent Document 2] Powers WJ, et al. 2018 Guidelines for the early management of patients with acute ischemic stroke: a guideline for healthcare professionals from the American Heart Association / American Stroke Association. Stroke. 2018; 49: e46- e99 [Non-Patent Document 3] Hemphill JC 3rd, et al. Guidelines for the management of spontaneous intracerebral hemorrhage: a guideline for healthcare professionals from the American Heart Association / American Stroke Association. Stroke. 2015; 46: 2032-2060 [Non-Patent Document 4] Savitz SI, et al. Reconsidering Neuroprotection in the Reperfusion Era. Stroke. 2017; 48: 3413-3419 [Non-Patent Document 5] Patel RAG, et al. Neuroprotection in the Treatment of Acute Ischemic Stroke. Prog Cardiovasc Dis. 2017; 59: 542-548 [Non-patent document 6] Roth GA, et al. Global, regional, and national age-sex-specific mortality for 282 causes of death in 195 countries and territories, 1980-2017: a systematic analysis for the Global Burden of Disease Study 2017. Lancet. 2018; 392:1736-1788 [Non-Patent Document 7] Marin B, et al. Variation in worldwide incidence of amyotrophic lateral sclerosis: a meta-analysis. Int J Epidemiol 2017; 46: 57-74 [Non-patent document 8] Rawlins MD. The Prevalence of Huntington's Disease. Neuroepidemiology. 2016; 46: 144-53 [Non-Patent Document 9] Battistin L, Cagnin A (December 2010). "Vascular cognitive disorder. A biological and clinical overview". Neurochemical Research. 35 (12): 1933-8. doi:10.1007 / s11064-010-0346-5. PMID 21127967 [Non-Patent Document 10] Plassman BL, Langa KM, Fisher GG, Heeringa SG, Weir DR, Ofstedal MB, Burke JR, Hurd MD, Potter GG, Rodgers WL, Steffens DC, Willis RJ, Wallace RB (2007). "Prevalence of dementia in the United States: the aging, demographics, and memory study". Neuroepidemiology. 29 (1-2): 125-32. doi:10.1159 / 000109998. PMC 2705925. PMID 17975326 [Non-Patent Document 11] Jorm AF, Korten AE, Henderson AS (November 1987). "The prevalence of dementia: a quantitative integration of the literature". Acta Psychiatrica Scandinavica. 76 (5): 465-79. doi:10.1111 / j.1600-0447.1987.tb02906.x. PMID 3324647 [Non-Patent Document 12] Atri A. The Alzheimer's Disease Clinical Spectrum: Diagnosis and Management. Med Clin North Am. 2019; 103: 263-293 [Non-Patent Document 13] Chaudhuri KR, et al. Unmet needs in Parkinson's disease: New horizons in a changing landscape. Parkinsonism Relat Disord. 2016; 33 (Suppl 1): S2-S8 Summary of the Invention [Problem to be solved by the invention]

[0008] Thus, there is a clear need for additional and better treatments that provide neuroprotection, particularly in the context of treating stroke, cerebral reperfusion injury, and certain neurodegenerative diseases. [Means for solving the problem]

[0009] The present invention provides a combination that is neuroprotective and suitable for the prevention or treatment of cerebral reperfusion injury, stroke, and other disorders / diseases where neuroprotection is desirable. Advantageously, the combinations and other aspects of the invention claimed herein provide treatments that are more effective and provide superior clinical outcomes compared to therapies using a single active pharmaceutical agent.

[0010] The first aspect is (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a combination comprising:

[0011] The second aspect is (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. The present invention relates to a combination comprising:

[0012] The third aspect is (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; a pharmaceutically acceptable carrier, diluent or excipient; The present invention relates to a pharmaceutical composition comprising:

[0013] The fourth aspect is (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof; a pharmaceutically acceptable carrier, diluent or excipient; The present invention relates to a pharmaceutical composition comprising:

[0014] The fifth aspect is (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a pharmaceutical product comprising:

[0015] The sixth aspect is (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. The present invention relates to a pharmaceutical product comprising:

[0016] A seventh aspect relates to a combination or pharmaceutical composition or pharmaceutical product as defined above for use in the treatment and / or prevention of one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative diseases, neonatal asphyxia, cardiac arrest, cardiogenic shock and acute myocardial infarction, or for use in providing cardioprotection against cardiotoxic drugs, or for use in providing neuroprotection, for example against neurotoxic drugs.

[0017] An eighth aspect relates to a pharmaceutical product as defined above for use in the treatment and / or prevention of one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative diseases, neonatal asphyxia, cardiac arrest, cardiogenic shock and acute myocardial infarction, or for use in providing cardioprotection against cardiotoxic drugs, or for use in providing neuroprotection, for example against neurotoxic drugs, wherein the components are for simultaneous, sequential or separate administration.

[0018] A ninth aspect relates to a method for treating and / or preventing one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection, for example against neurotoxic drugs, comprising administering to a subject in need thereof: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; simultaneously, sequentially or separately.

[0019] A tenth aspect relates to a method for treating and / or preventing one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection, for example against neurotoxic drugs, comprising administering to a subject in need thereof: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. simultaneously, sequentially or separately.

[0020] An eleventh aspect relates to the use of a compound of formula (I) or (II) in the manufacture of a medicament for treating and / or preventing one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection, for example against neurotoxic drugs, comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; Regarding the use of.

[0021] A twelfth aspect relates to the use of a compound of formula (I) or (II) in the manufacture of a medicament for treating and / or preventing one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection, for example against neurotoxic drugs, (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. Regarding the use of.

[0022] A thirteenth aspect relates to a method for treating and / or preventing ischemia and / or reperfusion injury in an ex vivo organ before or during transplantation, comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to the use of a combination comprising:

[0023] A fourteenth aspect relates to a method for treating and / or preventing ischemia and / or reperfusion injury in an ex vivo organ before or during transplantation, comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. The present invention relates to the use of a combination comprising: The present application also includes the following aspects. (1) (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; A combination including: (2) The combination according to (1) above, comprising a sulfonylurea and an aldosterone antagonist. (3) The combination according to (1) above, comprising a sulfonylurea and an insulin modulator. (4) The combination according to (1) above, comprising a sulfonylurea, an insulin modulator, and an aldosterone antagonist. (5) The combination according to any one of (1) to (4) above, wherein the insulin modulator is selected from exenatide and its structural and functional analogues, and pharmaceutically acceptable salts thereof. (6) The combination according to any one of (1) to (5) above, wherein the sulfonylurea is selected from glibenclamide and structural and functional analogues thereof. (7) The combination according to (5) above, wherein the structural or functional analogue of exenatide is a GLP-1 receptor agonist. (8) The combination according to (5) above, wherein the structural or functional analogue of exenatide is selected from lixisenatide, albiglutide, liraglutide, taspoglutide and dulaglutide (LY2189265). (9) The combination according to (6) above, wherein the structural or functional analogue of glibenclamide is selected from the acylhydrazone, sulfonamide and sulfonylthiourea derivatives of glibenclamide, glimepiride, glipizide and gliclazide, preferably gliclazide. (10) The combination according to any one of (1) to (9) above, wherein the aldosterone antagonist is selected from spironolactone, eplerenone, canrenone, potassium canrenoate, finerenone and prorenone, and, if applicable, pharmaceutically acceptable salts thereof. (11) The combination according to (10) above, wherein the aldosterone antagonist is potassium canrenoate or a structural or functional analog thereof. (12) The combination according to any one of the above (1) to (11), comprising at least one further active pharmaceutical ingredient (API) selected from beta blockers, renin-angiotensin inhibitors, statins (HMG-CoA reductase inhibitors), inhibitors of platelet activation or aggregation, phosphodiesterase-3 inhibitors, calcium sensitizers, antioxidants and anti-inflammatory agents. (13) A pharmaceutical composition comprising the combination according to any one of (1) to (12) above and a pharmaceutically acceptable carrier, diluent or excipient. (14) A pharmaceutical composition according to (13) above, in a form suitable for parenteral administration, preferably intravenous administration. (15) (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; Pharmaceutical products including: (16) (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. The pharmaceutical product according to (15) above, comprising: (17) The pharmaceutical product according to (16) above, comprising glibenclamide and exenatide, or a pharmaceutically acceptable salt thereof. (18) The pharmaceutical product according to (16) above, comprising glibenclamide and potassium canrenoate. (19) The pharmaceutical product according to (16) above, comprising glibenclamide, canrenoate potassium, and exenatide, or pharmaceutically acceptable salts thereof. (20) The combination according to any one of (1) to (12) above, or the pharmaceutical composition according to (13) or (14) above, for use in the treatment and / or prevention of one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for use in providing cardioprotection against cardiotoxic drugs, or for use in providing neuroprotection. (21) The combination or pharmaceutical composition for use according to (20) above, wherein the ischemia and / or reperfusion injury is cerebral, cardiac, pulmonary or renal ischemia and / or reperfusion injury, preferably cerebral ischemia, cerebral reperfusion injury or stroke. (22) The combination or pharmaceutical composition for use according to (20) or (21) above, wherein the components are for intravenous administration. (23) The combination or pharmaceutical composition for use according to any one of (20) to (22) above, wherein the components are for administration during reperfusion. (24) The combination or pharmaceutical composition for use according to any one of (20) to (22) above, wherein the components are for administration before reperfusion. (25) The combination or pharmaceutical composition for use according to any one of (20) to (22) above, wherein the components are for administration after reperfusion. (26) The pharmaceutical product according to any one of (15) to (19) above, wherein the components are for simultaneous, sequential, or separate administration, for use in the treatment and / or prevention of one or more of the following: ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for use in providing cardioprotection against cardiotoxic drugs, or for use in providing neuroprotection. (27) The pharmaceutical product for use according to (26) above, wherein the ischemia and / or reperfusion injury is cerebral, cardiac, pulmonary or renal ischemia and / or reperfusion injury, preferably cerebral ischemia, cerebral reperfusion injury or stroke. (28) (26) or (27) above, wherein the component is for parenteral administration, preferably for intravenous administration. 10. A pharmaceutical product for the use described in claim 1. (29) The pharmaceutical product for use according to any one of (26) to (28) above, wherein the component is for administration during reperfusion. (30) The pharmaceutical product for use according to any one of (26) to (28) above, wherein the component is for administration before reperfusion. (31) The pharmaceutical product for use according to any one of (26) to (28) above, wherein the component is for administration after reperfusion. (32) The pharmaceutical product for use according to any one of (26) to (31) above, wherein the ingredients are for simultaneous administration. (33) 1. A method for treating and / or preventing one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or providing cardioprotection against cardiotoxic drugs, or providing neuroprotection, comprising administering to a subject in need thereof: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The method comprises administering simultaneously, sequentially or separately (34) To those who need it, (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. The method according to (33) above, comprising simultaneously, sequentially or separately administering (35) The method according to (34) above, wherein the structural or functional analog of exenatide is a GLP-1 receptor agonist. (36) The method according to (34) or (35) above, wherein the structural or functional analogue of exenatide is selected from lixisenatide, albiglutide, liraglutide, taspoglutide and dulaglutide (LY2189265). (37) The method according to any one of the above (34) to (36), wherein the structural or functional analogue of glibenclamide is selected from acylhydrazone, sulfonamide and sulfonylthiourea derivatives of glibenclamide, glimepiride, glipizide and gliclazide, preferably gliclazide. (38) The method according to any one of (33) to (37) above, wherein the ischemia and / or reperfusion injury is ischemia and / or reperfusion injury of the brain, heart, lungs, or kidney, preferably cerebral ischemia, cerebral reperfusion injury, or stroke. (39) The method according to any one of the above (33) to (38), wherein the component is administered parenterally, preferably intravenously. (40) The method according to any one of (34) to (39) above, wherein glibenclamide is administered at a dose of about 0.001 to about 30 μg per kg of the subject's body weight. (41) The method according to any one of (34) to (39) above, wherein exenatide or a pharmaceutically acceptable salt thereof is administered at a dose of about 0.001 to about 1.5 μg per kg of the subject's body weight. (42) The method according to any one of (34) to (39) above, wherein potassium canrenoate is administered at a dose of about 0.03 to about 10 mg per kg of the subject's body weight. (43) The method according to any one of (33) to (42) above, which comprises simultaneously administering the components to a subject. (44) In the manufacture of a medicament for treating and / or preventing one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection, (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; Use of. (45) In the manufacture of a medicament for treating and / or preventing one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection, (a) glibenclamide, or a structural or functional analog thereof; (b) at least two of the following ingredients: (i) exenatide, or a structural or functional analogue thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analogue thereof. Use of. (46) The use according to (45) above, wherein the structural or functional analogue of exenatide is a GLP-1 receptor agonist. (47) The use according to (45) or (46) above, wherein the structural or functional analogue of exenatide is selected from lixisenatide, albiglutide, liraglutide, taspoglutide and dulaglutide (LY2189265). (48) The use according to any one of the above (45) to (47), wherein the structural or functional analogue of glibenclamide is selected from acylhydrazone, sulfonamide and sulfonylthiourea derivatives of glibenclamide, glimepiride, glipizide and gliclazide, preferably gliclazide. (49) The use according to any one of (44) to (48) above, wherein the ischemia and / or reperfusion injury is ischemia and / or reperfusion injury of the brain, heart, lungs, or kidney, preferably cerebral ischemia, cerebral reperfusion injury, or stroke. (50) The use according to any one of (44) to (49) above, wherein the component is administered parenterally, preferably intravenously. (51) The use according to any one of (44) to (50) above, wherein the component is administered during reperfusion. (52) The use according to any one of (44) to (50) above, wherein the component is administered before reperfusion. (53) The use according to any one of (44) to (50) above, wherein the component is administered after reperfusion. (54) The use according to any one of (44) to (53) above, which comprises administering the components simultaneously to a subject. (55) For treating and / or preventing ischemia and / or reperfusion injury in ex vivo organs prior to or during transplantation, (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; Use of combinations including: (56) For treating and / or preventing ischemia and / or reperfusion injury in ex vivo organs prior to or during transplantation, (a) glibenclamide, or a structural or functional analog thereof; (b) at least two of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. Use of combinations including: (57) The use according to (56) above, wherein the structural or functional analogue of exenatide is a GLP-1 receptor agonist. (58) The use according to (56) or (57) above, wherein the structural or functional analogue of exenatide is selected from lixisenatide, albiglutide, liraglutide, taspoglutide and dulaglutide (LY2189265). (59) The use according to any one of the above (56) to (58), wherein the structural or functional analogue of glibenclamide is selected from acylhydrazone, sulfonamide and sulfonylthiourea derivatives of glibenclamide, glimepiride and gliclazide, preferably gliclazide. (60) The use according to any one of (55) to (59) above, wherein the ischemia and / or reperfusion injury is cerebral ischemia, cerebral reperfusion injury, or stroke. (61) The use according to any one of (55) to (60) above, wherein the component is administered before transplantation. (62) The combination according to any one of (1) to (12) above, or the pharmaceutical composition according to (13) or (14) above, for use in the treatment and / or prevention of stroke. (63) The combination according to any one of (1) to (12) above, or the pharmaceutical composition according to (13) or (14) above, for use in the treatment and / or prevention of a neurodegenerative disease, preferably selected from Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, and vascular dementia. (64) A combination according to any one of (1) to (12) above, or a pharmaceutical composition according to (13) or (14) above, for use in providing neuroprotection. (65) The method according to any one of (33) to (43) above, wherein the neurodegenerative disease is selected from Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, and vascular dementia. DETAILED DESCRIPTION OF THE INVENTION

[0024] The preferred embodiments described below are applicable to any of the above aspects of the invention, as appropriate.

[0025] As used herein, a structural analog, also known as a chemical analog, is a compound that is similar to the structure of another compound but has a different structure with respect to certain components. A structural analog may differ in one or more atoms, functional groups, or substructures, which are replaced with other atoms, groups, or substructures. A structural analog can, at least theoretically, be imagined to be formed from another compound. A structural analog is often isoelectronic.

[0026] As used herein, a functional analog is a chemical compound that has similar physical, chemical, biochemical, or pharmacological properties as another compound. A functional analog is not necessarily a structural analog that has a similar chemical structure.

[0027] Sulfonylurea The combination of the present invention comprises a sulfonylurea as an essential ingredient.

[0028] Sulfonylureas are a class of oral hypoglycemic drugs primarily used in the management of type 2 diabetes and certain forms of monogenic diabetes. Sulfonylureas lower blood glucose levels by stimulating insulin secretion from the beta cells of the pancreas. Their primary target is the ATP-sensitive potassium (K) receptor in the beta cell plasma membrane. ATP) channel is a sulfonylurea receptor (SUR1) subunit (Proks P, et al. Diabetes. 2002; 51 (Suppl 3): S368-76, Gribble FM, Reimann F. Diabetologia. 2003; 46: 875-891).

[0029] Consistent with their introduction into the clinic, sulfonylureas are traditionally classified into two generations, differing primarily in their properties, with second-generation drugs allowing for less frequent administration (Sola D, et al. Arch Med Sci. 2015; 11: 840-8): First generation drugs include chlorpropamide, tolbutamide, acetohexamide, carbutamide, glycylamide, tolhexamide, metahexamide, and tolazamide; however, these are no longer used in clinical practice. Second-generation sulfonylureas include glibenclamide (glyburide), glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, and glyclopyramide. Modified / extended-release formulations are available for some second-generation sulfonylureas (gliclazide, glipizide).

[0030] Current guidelines recommend the use of second-generation sulfonylureas as second-line therapy in combination with metformin when metformin alone is inadequately controlled, and second-generation sulfonylureas may also be used in triple-drug combinations when adequate glycemic control is not achieved with the two-drug combination (Garber AJ, et al. Endocr Pract. 2019;25:69-100, Inzucchi SE, et al. Diabetes Care. 2015;38:140-9). The decision to use a sulfonylurea should take into account patient characteristics and potential adverse events associated with the sulfonylurea (Cordiner RLM, Pearson ER. Diabetes Obes Metab. 2019;21:761-771).

[0031] In one particularly preferred embodiment, the sulfonylurea is a Sur-1 receptor antagonist. Suitable Sur-1 receptor antagonists can be identified using known assays.

[0032] In one particularly preferred embodiment, the sulfonylurea is a SUR1-TRPM4 channel antagonist. Suitable SUR1-TRPM4 channel antagonists can be identified using known assays.

[0033] The present invention also encompasses structural or functional analogs of sulfonylureas, particularly those modified to extend the half-life of the drug, eg, conjugates of sulfonylureas.

[0034] In one preferred embodiment, the sulfonylurea is selected from glibenclamide (glyburide), glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, and glyclopyramide.

[0035] In one highly preferred embodiment, the sulfonylurea is selected from glibenclamide and its structural and functional analogs.

[0036] Preferably, the sulfonylurea is selected from the acylhydrazone, sulfonamide and sulfonylthiourea derivatives of glibenclamide, glimepiride, glipizide and gliclazide.

[0037] In one preferred embodiment, the sulfonylurea has the structure shown below:

[0038] [ka]

[0039] Glimepiride has the following structure:

[0040] In one preferred embodiment, the sulfonylurea has the structure shown below:

[0041] [ka]

[0042] Gliclazide has the formula:

[0043] In one preferred embodiment, the sulfonylurea has the structure shown below:

[0044] [ka]

[0045] Glipizide has the formula:

[0046] In one particularly preferred embodiment, the sulfonylurea is glibenclamide.

[0047] The systematic (IUPAC) name for glibenclamide is 5-chloro-N-[2-[4-(cyclohexylcarbamoyl-sulfamoyl)phenyl]ethyl]-2-methoxybenzamide (chemical formula C 23 H 28 ClN3O5S) with a molecular weight of 494; it has the following chemical structure:

[0048] [ka]

[0049] It has.

[0050] The present invention also encompasses structural and functional analogs of glibenclamide, particularly those modified to extend the half-life of the drug, such as conjugates of glibenclamide.

[0051] Glibenclamide (also known as glyburide) is a sulfonylurea receptor-1 (Sur-1) antagonist used as a hypoglycemic agent to treat diabetes. Glibenclamide has been investigated as a treatment to reduce edema after brain injuries such as ischemic stroke, traumatic brain injury, and subarachnoid hemorrhage, but results have been inconsistent (Wilkinson CM, et al. PLoS One. 2019;14:e0215952; Xu F, et al. Brain Behav. 2019;9(4):e01254; King ZA, et al. Drug Des Devel Ther. 2018;12:2539-2552). We investigated the role of glibenclamide as part of a combination therapy aimed at reducing reperfusion injury and potentially providing neuroprotection.

[0052] Glibenclamide is available generically and sold under many brand names, including Gliben-J, Daonil, Diabeta, Euglucon, Gilemal, Glidanil, Glybovin, Glynase, Maninil, Micronase, and Semi-Daonil, in doses of 1.25, 2.5, and 5 mg. Glibenclamide is used orally for the treatment of type 2 diabetes in tablet formulations (for adults) or oral suspensions (for children). The defined daily dose (DDD) of glibenclamide for the treatment of type 2 diabetes is 7 mg for the micronized formulation, which is highly bioavailable, and 10 mg for the conventional formulation. The defined daily dose (DDD) is the expected average daily maintenance dose of the drug used for its primary indication in adults, as determined according to the WHO Collaborating Centre for Drug Statistics Methodology. The DDD is a unit of measurement and does not necessarily reflect the recommended or prescribed daily dose. Treatment doses for individual patients and patient groups are based on individual characteristics (e.g., age, weight, ethnic differences, type and severity of disease) and pharmacokinetic considerations and therefore often differ from the DDD. The DDD values ​​for glibenclamide are obtained from the WHO Collaborating Centre for Drug Statistics Methodology (see https: / / www.whocc.no / atc_ddd_index / ?code=A10BB01&showdescription=yes). The usual starting dose of glibenclamide (micronized formulation) as initial therapy is 2.5 to 5 mg daily, and the usual maintenance dose ranges from 1.25 to 20 mg daily, which may be given as a single dose or in divided doses with breakfast or the first main meal (according to the FDA labeling for Micronase® glyburide tablets). This corresponds to a maintenance dose of approximately 18 μg / kg to approximately 285 μg / kg in a 70 kg adult.

[0053] Multiple studies in animal models have demonstrated a protective role for glibenclamide in inflammation-associated damage, including reducing harmful neuroinflammation and improving behavioral outcomes after central nervous system injury (Zhang G, et al. Mediators Inflamm. 2017; 2017: 3578702) or ischemic and hemorrhagic stroke (Caffes N, et al. Int J Mol Sci. 2015; 16: 4973-84). In a rat model of traumatic brain injury, glibenclamide was administered intraperitoneally as a 10 μg / kg loading dose followed by a 200 ng / hour infusion for 7 days (Patel AD, et al. J Neuropathol Exp Neurol. 2010;69:1177-90); whereas, in a mouse model of traumatic brain injury, the dose of glibenclamide was 10 μg for 3 days after controlled cortical impact injury (Xu ZM, et al. J Neurotrauma. 2017;34:925-933). In a rodent model of cerebral ischemia and reperfusion injury, glibenclamide was shown to be effective at a dose of 1 mg / kg administered 10 minutes before reperfusion (Abdallah DM, et al. Brain Res. 2011;1385:257-62). In rodent models of subarachnoid hemorrhage, glibenclamide was shown to be effective when administered intraperitoneally at a loading dose of 10 μg / kg followed by a 200 ng / h infusion for 24 hours (Simard, J. M, et al. Journal of Cerebral Blood Flow and Metabolism. 2009; 29; 317-330) or 1 week (Tosun C, et al. Stroke. 2013; 44: 3522-8). Glibenclamide administered as a continuous infusion (75 ng / h) reduced cerebral edema, infarct volume, and mortality by 50%, and the reduction in infarct volume was associated with cortical sparing 7 days after middle cerebral artery occlusion in a thromboembolic model of stroke in rats (Simard J. M, et al. Nat Med. 2006; 12: 433-40).Glibenclamide administered at a dose of 10 μg either before or 2 hours after experimental intracerebral hemorrhage in mice has been shown to reduce brain edema, BBB disruption, and neurological deficits (Xu F, et al. Brain Behav. 2019;9:e01254), and similar findings were obtained in another study (Jiang B, et al. Transl Stroke Res. 2017;8:183-193). However, a widely used dose of glibenclamide shown to be effective in other studies (10 μg / kg loading dose followed by 200 ng / hour for up to 7 days) was not shown to be effective when intracerebral hemorrhage was induced by intrastriatal injection of collagenase (Wilkinson CM, et al. PLoS One. 2019;14(5):e0215952).

[0054] Glibenclamide has also been shown to have beneficial effects in stroke patients in several clinical trials. The Glyburide Advantage in Malignant Edema and Stroke (GAMES) clinical trial, in which glyburide was administered intravenously as a 0.13 mg bolus intravenous injection over the first 2 minutes (RP-1127), followed by an infusion of 0.16 mg / hour for the first 6 hours and then 0.11 mg / hour for the remaining 66 hours, revealed promising findings regarding brain swelling (midline shift), MMP-9, functional outcomes, and mortality (King ZA, et al. Drug Des Devel Ther. 2018;12:2539-2552). An exploratory study of oral glibenclamide in patients with acute hemispheric infarction showed that treatment was safe, but 6-month functional outcomes did not substantially improve, although there was a slight trend toward less severe cerebral edema and less severe disability and death (Huang K, et al. Acta Neurol Scand. 2019 May 29). A retrospective analysis of data on sulfonylurea-naive diabetic patients in the days following acute ischemic stroke found a strong association between sulfonylurea treatment and improved survival, increased functional independence, reduced NIH Stroke Scale scores, and reduced hemorrhagic transformation (Kunte H, et al. Ann Neurol. 2012; 72: 799-806).

[0055] The above preclinical and clinical findings may be related to the upregulation of SUR1-TRPM4 channels after brain injury such as ischemia (Woo SK, et al. J Biol Chem. 2013; 288: 3655-67, Mehta RI et al. J Neuropathol Exp Neurol. 2015; 74: 835-49).

[0056] Neuroprotective effects in animal models of ischemia and reperfusion injury have also been reported for other sulfonylureas, such as gliclazide (Tan F, et al. Brain Res. 2014;1560: 83-90), and protective effects in animal models of ischemia and reperfusion injury in other tissues, such as the myocardium, for glimepiride (Nishida H et al. J Pharmacol Sci. 2009;109: 251-6).

[0057] Some other drugs have insulin secretagogue effects, such as sulfonylureas; examples include glinides (such as repaglinide, nateglinide, and mitiglinide). In addition, other compounds, such as resveratrol, bind to sulfonylurea receptors (Hambrock A, et al. J Biol Chem. 2007; 282: 3347-56) and have been shown to have neuroprotective effects in stroke and traumatic CNS injury (Lopez MS, et al. Neurochem Int. 2015; 89: 75-82).

[0058] Research by the applicant, and that is described in more detail in the accompanying Examples, has shown that administering a sulfonylurea (e.g., glibenclamide) in combination with a second active agent that is either an aldosterone antagonist (e.g., potassium canrenoate) or an insulin modulator (e.g., exenatide) provides a neuroprotective effect, even when the sulfonylurea is administered only in very low doses.

[0059] Insulin modulators In one embodiment, the combination of the present invention comprises, in addition to the sulfonylurea component described above, an insulin modulator.

[0060] As used herein, the term "insulin modulator" refers to an agent that can directly or indirectly increase or decrease the activity of insulin, which in turn can increase or decrease insulin-mediated physiological responses.

[0061] In one embodiment, the insulin modulator is selected from a GLP-1 agonist, a DPP-4 inhibitor, a PPAR agonist, insulin, and analogs thereof.

[0062] Examples of GLP-1 agonists include exenatide, lixisenatide, albiglutide, liraglutide, taspoglutide and dulaglutide (LY2189265) and pharmaceutically acceptable salts thereof.

[0063] Examples of DPP-4 inhibitors include sitagliptin, vildagliptin, saxagliptin, linagliptin, anagliptin, teneligliptin, alogliptin, trelagliptin, gemigliptin, dutogliptin, and omarigliptin (MK-3102), and pharmaceutically acceptable salts thereof.

[0064] Examples of PPAR agonists include clofibrate, gemfibrozil, ciprofibrate, bezafibrate, fenofibrate, saroglitazar, aleglitazar, muraglitazar and tesaglitazar, and pharmaceutically acceptable salts thereof.

[0065] Examples of insulin analogues include insulin lispro, insulin aspart, insulin glulisine, insulin detemir, insulin degludec, insulin glargine, and pharmaceutically acceptable salts thereof.

[0066] Thus, in one embodiment, the insulin modulator is selected from exenatide, lixisenatide, albiglutide, liraglutide, taspoglutide, dulaglutide (LY2189265), sitagliptin, vildagliptin, saxagliptin, linagliptin anagliptin, teneligliptin, alogliptin, trelagliptin, gemigliptin, dutogliptin, omarigliptin (MK-3102), clofibrate, gemfibrozil, ciprofibrate, bezafibrate, fenofibrate, saroglitazar, aleglitazar, muraglitazar, tesaglitazar, insulin lispro, insulin aspart, insulin glulisine, insulin detemir, insulin degludec, insulin glargine, and pharmaceutically acceptable salts thereof.

[0067] In one embodiment, the insulin modulator is a GLP-1 agonist selected from exenatide, lixisenatide, albiglutide, liraglutide, taspoglutide, dulaglutide (LY2189265), and pharmaceutically acceptable salts thereof. Preferably, the GLP-1 agonist is exenatide.

[0068] Exenatide In one preferred embodiment, the insulin modulator is selected from exenatide and its structural and functional analogs, and pharmaceutically acceptable salts thereof.

[0069] In one preferred embodiment, exenatide is in the form of a pharmaceutically acceptable salt, more preferably exenatide acetate. In another preferred embodiment, exenatide is in free base form.

[0070] As used herein, the term "exenatide" refers to a 39-mer peptide of the following sequence: H-His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Leu-Ser-Lys-Gln-Met-Glu-Glu-Glu-Ala-Val-Ar g-Leu-Phe-Ile-Glu-Trp-Leu-Lys-Asn-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2

[0071] Exenatide (synonymous exendin-4) was originally isolated from the saliva of the Gila monster Heloderma suspectum in 1992. It is an insulinotropic agent with glucoregulatory effects similar to those of the human peptide glucagon-like peptide-1 (GLP-1).

[0072] Exenatide mimics human glucagon-like peptide 1 (GLP-1), an intestinal incretin hormone released in response to nutrient intake (Goke R, et al., J. Biol. Chem., 1993, 268:19650-19655). It exerts insulinotropic and insulinomimetic properties via the GLP-1 receptor. GLP-1 receptors are widely expressed in many organs, including the heart and vascular endothelium (Bullock et al., Endocrinology, 1996, 137:2968-2978; Nystrom et al., Am J Physiol Endocrinol Metab, 2004, 287:E1209-E1215). Exenatide is currently approved as an antidiabetic drug for the treatment of patients with type 2 diabetes. The recommended dose for this indication is initially 5 micrograms (µg) twice daily, which is increased to 10 µg twice daily after 1 month based on clinical response.

[0073] GLP-1 is ineffective as a therapeutic agent because it has a very short circulating half-life (less than 2 minutes) due to rapid degradation by dipeptidyl peptidase-4. Exenatide is 50% homologous to GLP-1 but lacks the dipeptidyl peptidase-4 cleavage site, resulting in a half-life of 2.4 hours in humans.

[0074] Exenatide enhances glucose-dependent insulin secretion by pancreatic beta cells, suppresses inappropriately elevated glucagon secretion, and delays gastric emptying. Exenatide is highly potent, with a minimum effective concentration in humans of 50 pg / mL (12 pM). Current exenatide therapy involves twice-daily injections (Byetta®). A sustained-release formulation (Bydureon®) is also approved for weekly injections.

[0075] As used herein, a functional analog of exenatide refers to a compound that has a similar structure but differs therefrom in certain aspects (e.g., one or more atoms, functional groups, amino acid residues, or substructures may differ and be replaced by others). Functional analogs may exhibit similar pharmacological properties and may be structurally related.

[0076] In one embodiment, the structural or functional analog of exenatide is a form of exenatide that has been modified to extend its half-life, for example, a conjugate of exenatide.

[0077] In one preferred embodiment, the structural or functional analog of exenatide is PEGylated exenatide.For example, in one preferred embodiment, the structural or functional analog is exenatide monoPEGylated with 40kDa PEG.PEGylated exenatide can be prepared by methods known in the art.For example, PEGylated forms of exenatide are described in WO 2013 / 059323 (Prolynx LLC), the contents of which are incorporated herein by reference.Exenatide can also be conjugated to other molecules, such as proteins.

[0078] In one particularly preferred embodiment, the structural or functional analog of exenatide is in a sustained-release form, for example, sold under the trade name Bydureon®. In another preferred embodiment, the structural or functional analog of exenatide is in the form of multilayered nanoparticles for sustained delivery, for example, as described in (Kim JY, et al, Biomaterials, 2013; 34: 8444-9), the contents of which are incorporated herein by reference.

[0079] In another particularly preferred embodiment, exenatide is in an injectable form, such as that sold under the trade name Byetta®.

[0080] Functional analogues of exenatide include GLP receptor agonists. Suitable functional analogues of exenatide include lixisenatide, albiglutide, liraglutide, taspoglutide and dulaglutide (LY2189265).

[0081] In one embodiment, functional analogs of exenatide include modified exenatide in which one or more amino acid residues have been replaced with other amino acid residues, and / or one or more amino acid residues have been deleted, and / or one or more amino acid residues have been added and / or inserted.

[0082] In one embodiment, the functional exenatide analog contains fewer than 10 amino acid modifications (substitutions, deletions, additions (including insertions) and any combination thereof) compared to exenatide, or fewer than 9, 8, 7, 6, 5, 4, 3, or 2 modifications compared to exenatide.

[0083] In one embodiment, the functional exenatide analog contains 10 amino acid modifications (substitutions, deletions, additions (including insertions) and any combination thereof) compared to exenatide, or 9, 8, 7, 6, 5, 4, 3, 2 or 1 modification compared to exenatide.

[0084] Structural and functional analogs of exenatide also include salts, isomers, enantiomers, solvates, polymorphs, prodrugs, and metabolites thereof.

[0085] Aldosterone antagonists In one embodiment, the combination of the present invention comprises, in addition to the sulfonylurea component described above, an aldosterone antagonist.

[0086] Acute myocardial infarction and the subsequent hemodynamic changes lead to complex neurohormonal activation. The renin-angiotensin-aldosterone pathway is one basis of such neurohormonal activation. Aldosterone, which is at its highest level after acute myocardial infarction, has been reported to promote a wide range of adverse cardiovascular effects, including acute endothelial dysfunction, inhibition of nitric oxide activity, increased endothelial oxidative stress, increased vascular tone, inhibition of tissue recapture of catecholamines, rapid development of vascular smooth muscle cell and cardiomyocyte necrosis, intravascular collagen deposition, myocardial hypertrophy, and fibrosis (Struthers, Am Heart J, 2002, 144: S2-S7; Zannad and Radauceanu, Heart Fail Rev, 2005, 10: 71-78). Furthermore, it has been shown to predict poor outcomes (Beygui et al., Circulation, 2006, 114: 2604-2610).

[0087] Aldosterone antagonists, or antimineralcorticoids, are diuretics that antagonize the action of aldosterone at the mineralocorticoid receptor. This group of drugs is often used in the management of chronic heart failure. Members of this class are also used in the management of hyperaldosteronism (including Conn's syndrome) and hirsutism in women (due to additive antiandrogenic effects). Most antimineralcorticoids are steroidal spirolactones.

[0088] Mineralocorticoid receptor antagonism inhibits sodium absorption in the collecting ducts of the nephrons of the kidney. This interferes with sodium / potassium exchange, reduces urinary potassium excretion, and weakly increases water excretion (diuresis). In congestive heart failure, aldosterone antagonists are used in addition to other drugs for their additive diuretic effect, thereby reducing edema and cardiac workload.

[0089] Current guidelines recommend the use of mineralocorticoid receptor antagonists in patients with heart failure after myocardial infarction, based on the results of the EPHESUS trial.

[0090] Several studies in animal models of acute myocardial infarction and in the clinic have demonstrated the benefits of aldosterone blockade in preventing reperfusion injury and improving cardiac function in patients with STEMI. Literature has also shown that mineralocorticoid receptor antagonists may have beneficial effects on the cerebrovasculature and during stroke (Dinh QN, et al. Neural Regen Res. 2016; 11:1230-1).

[0091] Examples of aldosterone antagonists include spironolactone (the first and most widely used member of this class), eplerenone (much more selective for its target than spironolactone, but somewhat less potent and effective), canrenone and potassium canrenoate, finerenone (nonsteroidal and more potent and selective than eplerenone or spironolactone), and prorenone. Some drugs also have antimineralcorticoid effects secondary to their primary mechanism of action. Examples include progesterone, drospirenone, gestodene, and benidipine.

[0092] In one particularly preferred embodiment, the aldosterone antagonist is potassium canrenoate.

[0093] The present invention also encompasses structural and functional analogs of aldosterone antagonists, particularly those modified to extend the half-life of the drug, eg, conjugates of aldosterone antagonists.

[0094] Potassium canrenoate Canrenoate potassium, also known as the potassium salt of canrenoic acid, is an aldosterone antagonist of the spirolactone group. Like spironolactone, canrenoate potassium is a prodrug that is metabolized to canrenone in the body. Canrenoate potassium is typically administered intravenously at doses ranging from 200 mg / day to 600 mg / day for the treatment of hyperaldosteronism or hypokalemia.

[0095] Potassium canrenoate has the systematic (IUPAC) name potassium 3-[(8R,9S,10R,13S,14S,17R)-17-hydroxy-10,13-dimethyl-3-oxo-2,8,9,11,12,14,15,16-octahydro-1H-cyclopenta[a]phenanthren-17-yl]propanoate, formula C 22 H 29 KO4, with the following chemical structure:

[0096] [ka]

[0097] It has.

[0098] combination In one aspect, the present invention provides a method for producing a pharmaceutical composition comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a combination comprising:

[0099] The preferred embodiments described below apply mutatis mutandis to other aspects of the invention, including methods, uses, products and compositions.

[0100] In one preferred embodiment, the combination comprises a sulfonylurea and an insulin modulator.

[0101] In one preferred embodiment, the combination consists of a sulfonylurea and an insulin modulator.

[0102] In another preferred embodiment, the combination comprises a sulfonylurea and an aldosterone antagonist.

[0103] In another preferred embodiment, the combination consists of a sulfonylurea and an aldosterone antagonist.

[0104] In another preferred embodiment, the combination comprises a sulfonylurea, an insulin modulator, and an aldosterone antagonist.

[0105] In another preferred embodiment, the combination consists of a sulfonylurea, an insulin modulator, and an aldosterone antagonist.

[0106] In one embodiment, the insulin modulator is defined according to any of the above embodiments of an insulin modulator.

[0107] In one embodiment, the aldosterone antagonist is defined according to any of the above embodiments of an aldosterone antagonist.

[0108] In one embodiment, the sulfonylurea is defined according to any of the above embodiments of a sulfonylurea.

[0109] In one preferred embodiment, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof; and Preferably, the combination comprises (b)(i) and (b)(ii).

[0110] In one preferred embodiment, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. (a) at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, glyclopyramide, chlorpropamide, tolbutamide, acetohexamide, carbutamide, glyccyclamide, tolhexamide, metahexamide, and tolazamide; (b) the following components: (i) at least one of exenatide, lixisenatide, albiglutide, liraglutide, taspoglutide, and dulaglutide (LY2189265) or a pharmaceutically acceptable salt thereof; and (ii) at least one of potassium canrenoate, canrenone, spironolactone, eplerenone, finerenone, and prorenone, or, if applicable, a pharmaceutically acceptable salt thereof (e.g., a pharmaceutically acceptable salt of canrenone, spironolactone, eplerenone, finerenone, and prorenone); The present invention relates to a combination comprising:

[0111] In one embodiment, the present invention provides with at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, glyclopyramide, chlorpropamide, tolbutamide, acetohexamide, carbutamide, glyccyclamide, tolhexamide, metahexamide, and tolazamide; with at least one of exenatide, lixisenatide, albiglutide, liraglutide, taspoglutide, and dulaglutide (LY2189265) or a pharmaceutically acceptable salt thereof; At least one of canrenoate potassium, canrenone, spironolactone, eplerenone, finerenone, and prorenone, or, where applicable, a pharmaceutically acceptable salt thereof. The present invention relates to a combination comprising:

[0112] In one embodiment, the present invention provides with at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, glyclopyramide, chlorpropamide, tolbutamide, acetohexamide, carbutamide, glyccyclamide, tolhexamide, metahexamide, and tolazamide; At least one of exenatide, lixisenatide, albiglutide, liraglutide, taspoglutide, and dulaglutide (LY2189265) or a pharmaceutically acceptable salt thereof The present invention relates to a combination comprising:

[0113] In another embodiment, the present invention provides with at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, glyclopyramide, chlorpropamide, tolbutamide, acetohexamide, carbutamide, glyccyclamide, tolhexamide, metahexamide, and tolazamide; At least one of canrenoate potassium, canrenone, spironolactone, eplerenone, finerenone, and prorenone, or, where applicable, a pharmaceutically acceptable salt thereof. The present invention relates to a combination comprising:

[0114] In one embodiment, the present invention provides Exenatide or a pharmaceutically acceptable salt thereof; at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, glyclopyramide, chlorpropamide, tolbutamide, acetohexamide, carbutamide, glyccyclamide, tolhexamide, metahexamide, and tolazamide; or a combination comprising these.

[0115] In one embodiment, the present invention provides Exenatide or a pharmaceutically acceptable salt thereof; at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, and glyclopyramide; or a combination comprising these.

[0116] In one embodiment, the present invention provides Exenatide or a pharmaceutically acceptable salt thereof; Glibenclamide and or a combination comprising these.

[0117] In one embodiment, the present invention provides at least one of exenatide, lixisenatide, albiglutide, liraglutide, taspoglutide, and dulaglutide (LY2189265) or a pharmaceutically acceptable salt thereof; Glibenclamide and or a combination comprising these.

[0118] In one embodiment, the present invention provides potassium canrenoate, at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, glyclopyramide, chlorpropamide, tolbutamide, acetohexamide, carbutamide, glyccyclamide, tolhexamide, metahexamide, and tolazamide; or a combination comprising these.

[0119] In one embodiment, the present invention provides potassium canrenoate, at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, and glyclopyramide; or a combination comprising these.

[0120] In one embodiment, the present invention provides at least one of canrenoate potassium, canrenone, spironolactone, eplerenone, finerenone, and prorenone, or, where applicable, a pharmaceutically acceptable salt thereof; Glibenclamide and or a combination comprising these.

[0121] In one embodiment, the present invention provides Canrenoate potassium and; Glibenclamide and or a combination comprising these.

[0122] In one embodiment, the present invention provides with glibenclamide; with at least one of exenatide, lixisenatide, albiglutide, liraglutide, taspoglutide, and dulaglutide (LY2189265) or a pharmaceutically acceptable salt thereof; At least one of canrenoate potassium, canrenone, spironolactone, eplerenone, finerenone, and prorenone, or, where applicable, a pharmaceutically acceptable salt thereof. The present invention relates to a combination comprising:

[0123] In one embodiment, the present invention provides with at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, glyclopyramide, chlorpropamide, tolbutamide, acetohexamide, carbutamide, glyccyclamide, tolhexamide, metahexamide, and tolazamide; Exenatide or a pharmaceutically acceptable salt thereof; At least one of canrenoate potassium, canrenone, spironolactone, eplerenone, finerenone, and prorenone, or, where applicable, a pharmaceutically acceptable salt thereof. The present invention relates to a combination comprising:

[0124] In one embodiment, the present invention provides with at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, glyclopyramide, chlorpropamide, tolbutamide, acetohexamide, carbutamide, glyccyclamide, tolhexamide, metahexamide, and tolazamide; with at least one of exenatide, lixisenatide, albiglutide, liraglutide, taspoglutide, and dulaglutide (LY2189265) or a pharmaceutically acceptable salt thereof; Canrenoate potassium or canrenone The present invention relates to a combination comprising:

[0125] In one embodiment, the present invention provides Exenatide or a pharmaceutically acceptable salt thereof; Canrenoate potassium and; Glibenclamide and or a combination comprising these.

[0126] In one aspect of the invention, for each of the above embodiments, the combination consists of a sulfonylurea and an aldosterone antagonist and / or insulin modulator, i.e., these are the only active agents present. In another (alternative) aspect, the combination further comprises one or more additional active agents, as described below.

[0127] The effects of drug combinations are inherently unpredictable, and in many cases, one drug tends to partially or completely inhibit the effects of the other. The present invention demonstrates that a combination comprising a sulfonylurea, such as glibenclamide or a structural or functional analog thereof, and at least one of (i) an insulin modulator, such as exenatide, or a structural or functional analog thereof or a pharmaceutically acceptable salt thereof, and (ii) an aldosterone antagonist, such as potassium canrenoate, or a structural or functional analog thereof, when administered simultaneously, separately, or sequentially, does not cause any significant or dramatic adverse interactions between the two drugs. The unexpected lack of such antagonistic interactions is important for the clinical use of the combination.

[0128] Furthermore, preferred combinations according to the present invention surprisingly exhibit an enhanced effect of the individual components such that the optimal doses of the drugs are lower than those recommended for the approved indications of these drugs and / or are also lower than those reported in the literature for reperfusion injury.

[0129] In one embodiment, a combination of active agents of the present invention provides enhanced efficacy compared to each agent administered alone.

[0130] Illustratively, Applicant's studies have shown that the preferred doses of glibenclamide that produce a synergistic effect in the context of the combinations claimed herein are significantly lower than doses previously reported in the literature for blood glucose lowering (e.g., diabetes mellitus). Indeed, the preferred doses of glibenclamide used in the combinations claimed herein are approximately 20-285 times lower than the recommended maintenance doses of glibenclamide (micronized formulations) for the treatment of diabetes mellitus (for micronized formulations of glibenclamide, the recommended daily maintenance dose is 1.25-20 mg, which corresponds to 18 μg / kg-285 μg / kg for a 70 kg adult, in contrast to the preferred doses of glibenclamide as low as 1 μg / kg body weight required in the combination therapies claimed herein). Advantageously, using glibenclamide at these preferred low doses avoids effects on blood glucose levels that could otherwise lead to adverse side effects. Studies by the applicant have also shown that the clinically effective dose of glibenclamide as a double or triple combination according to the present invention with low doses of exenatide and / or potassium carbonate is also significantly lower than the doses of glibenclamide shown to be neuroprotective in published clinical studies (0.16 or 0.11 mg / hour continuous infusion, i.e., 3.84 mg or 2.64 mg per day) (see King ZA, et al.).

[0131] Furthermore, in another embodiment, the combination of active agents of the present invention provides unexpected synergistic effects, for example, in the treatment and / or prevention of reperfusion injury, particularly cerebral or myocardial reperfusion injury.

[0132] Combinations of two or more drugs can result in different types of drug interactions. A drug interaction is said to be additive if the combined effect of the two drugs is equal to the sum of the effects of each drug given alone. A drug interaction is said to be synergistic if the combined effect of the two drugs exceeds the effect of each drug given alone (Goodman and Gilmans "The Pharmacological Basis of Therapeutics", 12th Edition).

[0133] Combination therapy is an important treatment modality in many disease settings, including cardiovascular disease, cancer, and infectious diseases. Recent scientific advances have increased our understanding of the pathophysiological processes underlying these and other complex diseases. This increased understanding has further spurred the development of new therapeutic approaches that use drug combinations that target multiple therapeutic targets to improve treatment response, minimize the development of resistance, or minimize adverse events. In settings where combination therapy offers significant therapeutic benefits, there is growing interest in developing new combinations of two or more drugs.

[0134] Advantageously, a synergistic combination allows for lower doses of each component to be present, thereby reducing the toxicity of the therapy while producing and / or maintaining the same or an improved therapeutic effect. Thus, in particularly preferred embodiments, each component of the combination is present in a sub-therapeutic amount. The term "sub-therapeutically effective amount" means an amount lower than the amount typically required to produce a therapeutic effect for treatment with each agent alone.

[0135] In one embodiment, the present invention relates to a synergistic combination comprising a sulfonylurea and an insulin modulator.

[0136] In another embodiment, the present invention relates to a synergistic combination comprising a sulfonylurea and an aldosterone antagonist.

[0137] In another embodiment, the present invention relates to a synergistic combination comprising a sulfonylurea, an insulin modulator and an aldosterone antagonist.

[0138] In one embodiment, the insulin modulator is defined according to any of the above embodiments of an insulin modulator.

[0139] In one embodiment, the aldosterone antagonist is defined according to any of the above embodiments of an aldosterone antagonist.

[0140] In one embodiment, the sulfonylurea is defined according to any of the above embodiments of a sulfonylurea.

[0141] In one embodiment, the present invention relates to a synergistic combination comprising a sulfonylurea and at least one of exenatide, lixisenatide, albiglutide, liraglutide, taspoglutide, and dulaglutide (LY2189265), or a pharmaceutically acceptable salt thereof.

[0142] In one embodiment, the present invention relates to a synergistic combination comprising a sulfonylurea and at least one of canrenoate potassium, canrenone, spironolactone, eplerenone, finerenone, and prorenone, or, where applicable, a pharmaceutically acceptable salt thereof.

[0143] In one embodiment, the present invention provides with at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, and glyclopyramide; and at least one of exenatide or a pharmaceutically acceptable salt thereof. Canrenoate potassium and The present invention relates to a synergistic combination comprising:

[0144] In one embodiment, the present invention provides with at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, and glyclopyramide; with at least one of exenatide, lixisenatide, albiglutide, liraglutide, taspoglutide, and dulaglutide (LY2189265) or a pharmaceutically acceptable salt thereof; Canrenoate potassium and The present invention relates to a synergistic combination comprising:

[0145] In one embodiment, the present invention provides with at least one of exenatide, lixisenatide, albiglutide, liraglutide, taspoglutide, and dulaglutide (LY2189265) or a pharmaceutically acceptable salt thereof; Glibenclamide and The present invention relates to a synergistic combination comprising:

[0146] In one embodiment, the present invention provides Exenatide or a pharmaceutically acceptable salt thereof, Glibenclamide and The present invention relates to a synergistic combination comprising:

[0147] In one embodiment, the present invention provides Exenatide or a pharmaceutically acceptable salt thereof; at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, and glyclopyramide; The present invention relates to a synergistic combination comprising:

[0148] In one embodiment, the present invention provides with at least one of canrenoate potassium, canrenone, spironolactone, eplerenone, finerenone and prorenone, or a pharmaceutically acceptable salt thereof; Glibenclamide and The present invention relates to a synergistic combination comprising:

[0149] In one embodiment, the present invention provides Canrenoate potassium; and and at least one of glibenclamide, glibornuride, gliclazide, glipizide, glimepiride, gliquidone, glisoxepide, and glyclopyramide.

[0150] In one embodiment, the present invention relates to a synergistic combination comprising canrenoate potassium and glibenclamide.

[0151] In one embodiment, the present invention relates to a synergistic combination comprising exenatide or a pharmaceutically acceptable salt thereof, potassium canrenoate, and glibenclamide.

[0152] Additional Active Pharmaceutical Ingredients In one embodiment, the combination described above comprises at least one additional active pharmaceutical ingredient (API).

[0153] In one embodiment, the combination described above may further comprise at least one additional API selected from beta blockers, renin-angiotensin inhibitors, statins (HMG-CoA reductase inhibitors), inhibitors of platelet activation or aggregation, phosphodiesterase-3 inhibitors, calcium sensitizers, antioxidants, and anti-inflammatory agents.

[0154] Examples of beta blockers include propranolol, metoprolol, bucindolol, carteolol, carvedilol, labetalol, nadolol, oxprenolol, penbutolol, pindolol, sotalol, and timolol.

[0155] Renin-angiotensin inhibitors include angiotensin converting enzyme inhibitors, angiotensin AT1 receptor inhibitors and renin inhibitors.

[0156] Examples of angiotensin converting enzyme inhibitors include captopril, zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trandolapril, cilazapril, and fosinopril.

[0157] Examples of angiotensin AT1 receptor antagonists include losartan, irbesartan, olmesartan, candesartan, valsartan, fimasartan and telmisartan.

[0158] Examples of renin inhibitors include remikiren and aliskiren.

[0159] Examples of calcium sensitizers include levosimendan and its analogs.

[0160] Examples of statins include atorvastatin, lovastatin, pravastatin, rosuvastatin, and simvastatin.

[0161] Examples of platelet activation or aggregation inhibitors include prostacyclin (epoprostenol) and its structural and functional analogs (e.g., treprostinil, iloprost), irreversible cyclooxygenase inhibitors (e.g., aspirin, triflusal), adenosine diphosphate (ADP) receptor inhibitors (e.g., clopidogrel, prasugrel, ticagrelor, ticlopidine), phosphodiesterase inhibitors (e.g., cilostazol), protease-activated receptor-1 (PAR-1) antagonists (e.g., vorapaxar), glycoprotein IIB / IIIA inhibitors (e.g., abciximab, eptifibatide, tirofiban), adenosine reuptake inhibitors (e.g., dipyridamole), and thromboxane inhibitors, including thromboxane synthase inhibitors and thromboxane receptor antagonists (e.g., terutroban).

[0162] Examples of phosphodiesterase-3 (PDE-3) inhibitors include amrinone, milrinone, and analogs thereof.

[0163] Examples of antioxidants include ascorbic acid, lipoic acid, glutathione, melatonin, and resveratrol.

[0164] Examples of anti-inflammatory agents include COX-2 inhibitors (eg, celecoxib), glucocorticoids (eg, hydrocortisone), and non-steroidal anti-inflammatory drugs (eg, ibuprofen).

[0165] In one embodiment, the combination comprises at least one additional API selected from propranolol, metoprolol, bucindolol, carteolol, carvedilol, labetalol, nadolol, oxprenolol, penbutolol, pindolol, sotalol, timolol, captopril, zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trandolapril, cilazapril, fosinopril, losartan, irbesartan, olmesartan, candesartan, valsartan, fimasartan, telmisartan, remikiren, aliskiren, melatonin, and resveratrol.

[0166] In another embodiment, the above combination comprises at least one additional API selected from carvedilol, metoprolol, losartan, irbesartan, olmesartan, candesartan, valsartan, fimasartan, telmisartan, captopril, zofenopril, enalapril, ramipril, quinapril, perindopril, lisinopril, benazepril, imidapril, trandolapril, cilazapril, fosinopril, remikiren, aliskiren, melatonin, and resveratrol.

[0167] In another embodiment, the above combination comprises at least one additional API selected from carvedilol, metoprolol, melatonin, and resveratrol.

[0168] pharmaceutically acceptable salts The active pharmaceutical agents of the present invention can be present as pharmaceutically acceptable salts.

[0169] Pharmaceutically acceptable salts of the agents of the present invention include suitable acid addition or base salts thereof. A review of suitable pharmaceutical salts can be found in (Berge et al., J Pharm Sci, 66, 1-19 (1977)). Salts are formed, for example, with strong inorganic acids, such as mineral acids, for example sulfuric acid, phosphoric acid or hydrohalic acids (for example HCl, HBr); with strong organic carboxylic acids, for example alkanecarboxylic acids of 1 to 4 carbon atoms, which are unsubstituted or substituted (for example by halogens), for example acetic acid; with saturated or unsaturated dicarboxylic acids, for example oxalic acid, malonic acid, succinic acid, maleic acid, fumaric acid, phthalic acid or tetraphthalic acid; with hydroxycarboxylic acids, for example ascorbic acid, glycolic acid, lactic acid, malic acid, tartaric acid or citric acid; with amino acids, for example aspartic acid or glutamic acid; with benzoic acid; or with organic sulfonic acids, for example (C1-C4)-alkylsulfonic acids or arylsulfonic acids, which are unsubstituted or substituted (for example by halogens), for example methanesulfonic acid or p-toluenesulfonic acid.

[0170] Enantiomers / Tautomers The present invention also includes, where appropriate, all enantiomers and tautomers of the active pharmaceutical agents. Those skilled in the art will recognize compounds that have optical properties (one or more chiral carbon atoms) or tautomeric characteristics. The corresponding enantiomers and / or tautomers can be isolated / prepared by methods known in the art.

[0171] Stereoisomers and geometric isomers Some of the active pharmaceutical agents of the present invention may exist as stereoisomers and / or geometric isomers, e.g., they may have one or more asymmetric and / or geometric centers and therefore may exist in two or more stereoisomeric and / or geometric forms. The present invention contemplates the use of all individual stereoisomers and geometric isomers of these inhibitors, as well as mixtures thereof. The terms used in the claims encompass these forms, so long as they retain the appropriate functional activity (though not necessarily to the same degree).

[0172] The present invention also encompasses all suitable isotopic variations of the active pharmaceutical agent or its pharmaceutically acceptable salt. An isotopic variation of an agent of the present invention or its pharmaceutically acceptable salt is defined as one in which at least one atom is replaced by an atom having the same atomic number but an atomic mass different from that normally found in nature. Examples of isotopes that can be incorporated into the agent and its pharmaceutically acceptable salt include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as 2H, 3H, 13C, 14C, 15N, 17O, 18O, 31P, 32P, 35S, 18F, and 36Cl, respectively. Certain isotopic variations of the agent and its pharmaceutically acceptable salt, for example, those incorporating radioactive isotopes such as 3H or 14C, are useful in drug and / or substrate tissue distribution studies. Tritiated, i.e., 3H, and carbon-14, i.e., 14C, isotopes are particularly preferred due to their ease of preparation and detectability. Additionally, substitution with isotopes such as deuterium, i.e., 2H, can afford certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and therefore may be preferable in some circumstances. Isotopic variations of the agents of the present invention and pharmaceutically acceptable salts thereof of the present invention can generally be prepared by conventional procedures using appropriate isotopic variations of suitable reagents.

[0173] solvate The present invention also includes solvated forms of the active pharmaceutical agents of the invention, and the terms used in the claims encompass these forms.

[0174] polymorph The present invention further relates to the active pharmaceutical agents of the present invention in various crystalline, polymorphic and (an)hydrated forms. It is well established within the pharmaceutical industry that chemical compounds can be isolated in any of these forms by slight variations in the purification and / or isolation methods from solvents used in the synthetic preparation of such compounds.

[0175] Pharmaceutical Composition In another aspect, the present invention relates to a pharmaceutical composition comprising a combination according to the invention as described above and a pharmaceutically acceptable carrier, diluent or excipient.

[0176] In one aspect, the present invention provides a method for producing a pharmaceutical composition comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; a pharmaceutically acceptable carrier, diluent or excipient; The present invention relates to a pharmaceutical composition comprising:

[0177] In one preferred embodiment, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. (a) glibenclamide, or a structural or functional analog thereof; and a pharmaceutically acceptable carrier, diluent, or excipient; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof; a pharmaceutically acceptable carrier, diluent or excipient; The present invention relates to a pharmaceutical composition comprising:

[0178] In one aspect, the present invention provides a method for producing a pharmaceutical composition comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; and a pharmaceutically acceptable carrier, diluent or excipient.

[0179] In one preferred embodiment, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. (a) glibenclamide, or a structural or functional analog thereof; and a pharmaceutically acceptable carrier, diluent, or excipient; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof; a pharmaceutically acceptable carrier, diluent or excipient; The present invention relates to a pharmaceutical composition comprising:

[0180] While the compounds of the invention (including their pharmaceutically acceptable salts) can be administered alone, they will generally be administered in admixture with a pharmaceutical carrier, excipient, or diluent, particularly for human therapy. The pharmaceutical compositions may be for human or non-human animal use, in human and veterinary medicine, respectively.

[0181] Examples of such excipients suitable for the various different forms of pharmaceutical compositions described herein can be found in "Handbook of Pharmaceutical Excipients", 2nd Edition, (1994), edited by A Wade and PJ Weller.

[0182] Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical art and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (AR Gennaro edit. 1985).

[0183] The choice of pharmaceutical carrier, excipient, or diluent can be selected with regard to the intended route of administration and standard pharmaceutical practice. Exemplary routes of administration include parenteral (e.g., intravenous, intramuscular, intradermal, intraperitoneal, or subcutaneous), oral, inhalation, transdermal (topical), intraocular, iontophoretic, and transmucosal administration.

[0184] In one embodiment, the pharmaceutical composition is for parenteral administration (e.g., intravenous, intraarterial, intrathecal, intramuscular, intradermal, intraperitoneal, or subcutaneous). Preferably, the composition is prepared from a sterile or sterilizable solution.

[0185] In another embodiment, the pharmaceutical composition is for intravenous, intramuscular, or subcutaneous administration.

[0186] In another embodiment, the pharmaceutical composition is for intravenous administration.

[0187] Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline, fixed oils, polyethylene glycol, glycerin, propylene glycol, or other synthetic solvents; an antibacterial agent such as benzyl alcohol or methylparaben; an antioxidant such as ascorbic acid or sodium bisulfite; a chelating agent such as ethylenediaminetetraacetic acid; a buffer such as acetate, citrate, or phosphate, and an agent for adjusting isotonicity such as sodium chloride or dextrose. pH can be adjusted with acids or bases such as hydrochloric acid or sodium hydroxide.

[0188] Pharmaceutical compositions suitable for injectable use can include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor™, or phosphate buffered saline (PBS). In all cases, a composition for parenteral administration must be sterile and should be fluid to the extent that easy injectability with a syringe is possible. It should be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms, such as bacteria and fungi.

[0189] Sterile injectable solutions can be prepared by incorporating the required amount of active compound into a suitable solvent with one or a combination of the ingredients listed above, as needed, followed by filtration sterilization.Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle containing a basic dispersion medium and the other necessary ingredients listed above.For the preparation of sterile injectable powders, typical preparation methods include vacuum drying and freeze-drying, which can produce powders of the active ingredient and any additional desired ingredients from the solution that has previously been sterile-filtered.The present invention also encompasses liposome and nanoparticle preparations containing active agents.Such preparations, together with the methods for their preparation, are well known to those skilled in the art.

[0190] Pharmaceutical products In another aspect, the present invention provides a method for producing a pharmaceutical composition comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a pharmaceutical product comprising:

[0191] In one preferred embodiment, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. The present invention relates to a pharmaceutical product comprising:

[0192] In another aspect, the present invention provides a method for producing a pharmaceutical composition comprising: (a) with sulfonylureas; (b) at least one of the following ingredients: (i) an insulin modulator, and (ii) an aldosterone antagonist.

[0193] In one preferred embodiment, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. (a) glibenclamide, or a structural or functional analog thereof; (b) A pharmaceutical product comprising at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) potassium canrenoate, or a structural or functional analog thereof.

[0194] In one preferred embodiment, each component of the pharmaceutical product is for separate administration.

[0195] In one embodiment, the pharmaceutical product is a kit of parts containing all the supplies needed for the course of treatment (eg, drug vials, diluent vials, syringes, and needles).

[0196] The components of the kit and pharmaceutical product are as described above. In a preferred embodiment, each component of the kit or pharmaceutical product is admixed with one or more pharmaceutically acceptable diluents, excipients, and / or carriers.

[0197] In one embodiment, the kit includes a separate container for each active agent, which may be an ampule, a disposable syringe, or a multi-dose vial.

[0198] In another embodiment, the kit comprises a container containing a combined preparation of each active agent.

[0199] The kit may further comprise instructions for the treatment and / or prevention of reperfusion injury.

[0200] medical use The present invention further relates to the use of the above combinations, pharmaceutical products or pharmaceutical compositions in treating various therapeutic disorders, as detailed below, and related methods of treatment.

[0201] In one preferred embodiment, each of the pharmaceutically active ingredients of the combination, pharmaceutical product or pharmaceutical composition is administered separately.

[0202] In one aspect, the present invention provides a method for the treatment and / or prevention of one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection, for example, against neurotoxic drugs. (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a combination comprising:

[0203] In one preferred embodiment, the combination is for use in providing neuroprotection. Preferably, the combination is for use in providing neuroprotection against neurotoxic drugs.

[0204] In one preferred embodiment, the present invention provides a method for producing a medicament for the treatment of a pulmonary arthritis. for use in the treatment and / or prevention of one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for use in providing cardioprotection against cardiotoxic drugs, or for use in providing neuroprotection against neurotoxic drugs; (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. The present invention relates to a combination comprising:

[0205] In another aspect, the present invention provides a method for the treatment and / or prevention of one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection against neurotoxic drugs. (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a pharmaceutical composition comprising a combination comprising:

[0206] In one preferred embodiment, the present invention provides a method for the treatment and / or prevention of one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection against neurotoxic drugs. (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; (ii) canrenoate potassium, or a structural or functional analog thereof; The present invention relates to a pharmaceutical composition comprising a combination comprising:

[0207] In another aspect, the present invention provides a method for the treatment and / or prevention of one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection against neurotoxic drugs. (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; A pharmaceutical product comprising: The components relate to pharmaceutical products that are for simultaneous, sequential or separate administration.

[0208] In one preferred embodiment, the present invention provides a method for the treatment and / or prevention of one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection against neurotoxic drugs. (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. A pharmaceutical product comprising: The components relate to pharmaceutical products that are for simultaneous, sequential or separate administration.

[0209] In another aspect, the present invention provides a method for the manufacture of a medicament for treating and / or preventing one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection against neurotoxic drugs, comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; Regarding the use of.

[0210] In another preferred embodiment, the present invention relates to a method for the treatment and / or prevention of one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection against neurotoxic drugs, comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. Regarding the use of.

[0211] In one embodiment, the present invention provides a method for treating and / or preventing reperfusion injury comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a combination comprising:

[0212] In one preferred embodiment, the present invention provides a method for treating and / or preventing reperfusion injury, comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least two of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. The present invention relates to a combination comprising:

[0213] In another embodiment, the present invention provides a method for treating and / or preventing reperfusion injury comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a pharmaceutical composition comprising:

[0214] In another preferred embodiment, the present invention provides a method for treating and / or preventing reperfusion injury comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; (ii) canrenoate potassium, or a structural or functional analog thereof; The present invention relates to a pharmaceutical composition comprising a combination comprising:

[0215] In another embodiment, the present invention provides a method for treating and / or preventing reperfusion injury comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; A pharmaceutical product comprising: The components relate to pharmaceutical products that are for simultaneous, sequential or separate administration.

[0216] In another preferred embodiment, the present invention provides a method for treating and / or preventing reperfusion injury comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; (ii) canrenoate potassium, or a structural or functional analog thereof; A pharmaceutical product comprising: The components relate to pharmaceutical products that are for simultaneous, sequential or separate administration.

[0217] In another embodiment, the present invention provides a method for the manufacture of a medicament for the treatment and / or prevention of reperfusion injury, comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; Regarding the use of.

[0218] In another preferred embodiment, the present invention provides a method for the preparation of a medicament for the treatment and / or prevention of reperfusion injury, comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. Regarding the use of.

[0219] In one preferred embodiment, the present invention provides a method for the treatment and / or prevention of ischemia comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a combination comprising:

[0220] In one preferred embodiment, the present invention provides a method for the treatment and / or prevention of ischemia comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; (ii) canrenoate potassium, or a structural or functional analog thereof; The present invention relates to a combination comprising:

[0221] In another embodiment, the present invention provides a method for the treatment and / or prevention of ischemia comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a pharmaceutical composition comprising a combination comprising:

[0222] In another preferred embodiment, the present invention provides a method for the treatment and / or prevention of ischemia comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. The present invention relates to a pharmaceutical composition comprising a combination comprising:

[0223] In another embodiment, the present invention provides a method for the treatment and / or prevention of ischemia comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; A pharmaceutical product comprising: The components relate to pharmaceutical products that are for simultaneous, sequential or separate administration.

[0224] In another preferred embodiment, the present invention provides a method for the treatment and / or prevention of ischemia comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. A pharmaceutical product comprising: The components relate to pharmaceutical products that are for simultaneous, sequential or separate administration.

[0225] In another embodiment, the present invention provides a method for the manufacture of a medicament for the treatment and / or prevention of ischemia, comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; Regarding the use of.

[0226] In another preferred embodiment, the present invention provides a method for the manufacture of a medicament for the treatment and / or prevention of ischemia, comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; (ii) canrenoate potassium, or a structural or functional analog thereof; Regarding the use of.

[0227] In one preferred embodiment, the present invention provides a compound comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a combination comprising:

[0228] In one preferred embodiment, the present invention provides a compound comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; (ii) canrenoate potassium, or a structural or functional analog thereof; The present invention relates to a combination comprising:

[0229] In another embodiment, the present invention provides a compound comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a pharmaceutical composition comprising a combination comprising:

[0230] In another preferred embodiment, the present invention provides a method for the treatment and / or prevention of stroke comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. The present invention relates to a pharmaceutical composition comprising a combination comprising:

[0231] In another embodiment, the present invention provides a compound comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; A pharmaceutical product comprising: The components relate to pharmaceutical products that are for simultaneous, sequential or separate administration.

[0232] In another preferred embodiment, the present invention provides a method for the treatment and / or prevention of stroke comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. A pharmaceutical product comprising: The components relate to pharmaceutical products that are for simultaneous, sequential or separate administration.

[0233] In another embodiment, the present invention provides a method for the manufacture of a medicament for the treatment and / or prevention of stroke, comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; Regarding the use of.

[0234] In another preferred embodiment, the present invention provides a method for the manufacture of a medicament for the treatment and / or prevention of stroke, comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; (ii) canrenoate potassium, or a structural or functional analog thereof; Regarding the use of.

[0235] In one preferred embodiment, the stroke is a hemorrhagic stroke.

[0236] In another preferred embodiment, the stroke is an ischemic stroke.

[0237] As used herein, the term "reperfusion injury" refers to damage to tissues caused when blood supply returns to the tissue after a period of ischemia. The lack of oxygen and nutrients from the blood creates a condition in which restoration of circulation leads to inflammation, mitochondrial dysfunction, and oxidative damage through the induction of oxidative stress, rather than restoration of normal function. Reperfusion injury can occur after either a naturally occurring event, such as arterial occlusion, or a planned event, such as some surgical interventions. Myocardial reperfusion injury can occur, for example, after myocardial infarction or as a result of heart transplantation. Cerebral reperfusion injury can occur, for example, after ischemic stroke or as a result of neonatal asphyxia.

[0238] In one embodiment, the ischemia and / or reperfusion injury is ischemia and / or reperfusion injury of the brain, heart, lung, kidney or other organ / tissue susceptible to ischemia and / or reperfusion injury.

[0239] In one embodiment, the ischemia and / or reperfusion injury is cerebral ischemia and / or reperfusion injury, preferably cerebral ischemia and / or cerebral reperfusion injury.

[0240] In one embodiment, the ischemia and / or reperfusion injury is cardiac ischemia and / or reperfusion injury, preferably myocardial ischemia and / or myocardial reperfusion injury.

[0241] In one embodiment, the present invention provides a method for the treatment and / or prevention of a neurodegenerative disorder comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a combination comprising:

[0242] In one preferred embodiment, the present invention provides a compound comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least two of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. The present invention relates to a combination comprising:

[0243] In another embodiment, the present invention provides a compound comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to a pharmaceutical composition comprising:

[0244] In another preferred embodiment, the present invention provides a method for the treatment and / or prevention of neurodegenerative disorders comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; (ii) canrenoate potassium, or a structural or functional analog thereof; The present invention relates to a pharmaceutical composition comprising a combination comprising:

[0245] In another embodiment, the present invention provides a compound comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; A pharmaceutical product comprising: The components relate to pharmaceutical products that are for simultaneous, sequential or separate administration.

[0246] In another preferred embodiment, the present invention provides a method for the treatment and / or prevention of neurodegenerative disorders comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; (ii) canrenoate potassium, or a structural or functional analog thereof; A pharmaceutical product comprising: The components relate to pharmaceutical products that are for simultaneous, sequential or separate administration.

[0247] In another embodiment, the present invention provides a method for the manufacture of a medicament for the treatment and / or prevention of a neurodegenerative disorder, comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; Regarding the use of.

[0248] In another preferred embodiment, the present invention provides a method for the preparation of a medicament for the treatment and / or prevention of a neurodegenerative disorder, comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; (ii) canrenoate potassium, or a structural or functional analog thereof; Regarding the use of.

[0249] In one preferred embodiment, the neurodegenerative disorder is selected from Parkinson's disease, amyotrophic lateral sclerosis (ALS), Huntington's disease, and Alzheimer's disease.

[0250] In one preferred embodiment, the neurodegenerative disorder is Parkinson's disease.

[0251] In one preferred embodiment, the neurodegenerative disorder is amyotrophic lateral sclerosis (ALS).

[0252] In one preferred embodiment, the neurodegenerative disorder is vascular dementia.

[0253] In one preferred embodiment, the neurodegenerative disorder is Alzheimer's disease.

[0254] The insulin modulator, aldosterone antagonist, and sulfonylurea may be for simultaneous, sequential, or separate administration (as part of a dosing regimen).

[0255] Exenatide or a structural or functional analogue thereof or a pharmaceutically acceptable salt thereof, canrenoate potassium or a structural or functional analogue thereof, and glibenclamide or a structural or functional analogue thereof or a pharmaceutically acceptable salt thereof may be for simultaneous, sequential, or separate administration (as part of a dosing regimen).

[0256] As used herein, "concurrently" is used to mean that the two agents are administered simultaneously.

[0257] As used herein, "sequentially" means that the active agents are not administered simultaneously, but one is administered after the other. Thus, "sequential" administration may allow one agent to be administered within 5 minutes, 10 minutes, or approximately several hours after the other, as long as the circulating half-life of the agent administered first is such that both the agent and the other agent are present simultaneously in therapeutically effective amounts. The time delay between the administration of these components varies depending on the exact nature of these components, the interaction between them, and their respective half-lives. "Separately," as opposed to "sequentially," is used herein to mean that there is a significant gap between the administration of one agent and the other, i.e., the agent administered first may no longer be present in the bloodstream in therapeutically effective amounts when the second agent is administered.

[0258] In one embodiment, the components of the combination are for simultaneous administration.

[0259] Treatment method In another aspect, the present invention provides a method for treating and / or preventing ischemia and / or reperfusion injury, comprising administering to a subject: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; simultaneously, sequentially or separately.

[0260] In another preferred embodiment, the present invention provides a method for treating and / or preventing ischemia and / or reperfusion injury, comprising administering to a subject in need thereof: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. simultaneously, sequentially or separately.

[0261] In one embodiment, the present invention provides a method for treating and / or preventing reperfusion injury, comprising administering to a subject in need thereof: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; simultaneously, sequentially or separately.

[0262] In one preferred embodiment, the present invention provides a method for treating and / or preventing reperfusion injury, comprising administering to a subject in need thereof: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. simultaneously, sequentially or separately.

[0263] In another embodiment, the present invention provides a method for treating and / or preventing ischemia, comprising administering to a subject in need thereof: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; simultaneously, sequentially or separately.

[0264] In another embodiment, the present invention provides a method for treating and / or preventing ischemia, comprising administering to a subject in need thereof: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. simultaneously, sequentially or separately.

[0265] In one embodiment, the method relates to treating and / or preventing ischemia and / or reperfusion injury in the brain, heart, lungs, kidneys or other organs / tissues susceptible to ischemia and / or reperfusion injury.

[0266] In one embodiment, the method relates to treating and / or preventing reperfusion injury in the brain, heart, lungs, kidneys or other organs / tissues susceptible to reperfusion injury.

[0267] In one embodiment, the method relates to treating and / or preventing ischemia in the brain, heart, lungs, kidneys or other organs / tissues prone to ischemia.

[0268] In another embodiment, the method relates to treating and / or preventing cerebral ischemia and / or reperfusion injury, preferably cerebral ischemia and / or cerebral reperfusion injury.

[0269] In another embodiment, the method relates to treating and / or preventing cerebral reperfusion injury, preferably cerebral reperfusion injury.

[0270] In another embodiment, the method relates to treating and / or preventing cardiac ischemia and / or reperfusion injury, preferably myocardial ischemia and / or myocardial reperfusion injury.

[0271] In another embodiment, the method relates to treating and / or preventing cardiac reperfusion injury, preferably myocardial reperfusion injury.

[0272] In one particularly preferred embodiment, the method relates to treating and / or preventing acute myocardial infarction, which is one of the most common clinical indications of reperfusion injury.

[0273] In one particularly preferred embodiment, the method relates to treating and / or preventing stroke.

[0274] In one preferred embodiment, the stroke is a hemorrhagic stroke.

[0275] In one particularly preferred embodiment, the method relates to treating and / or preventing ischemic stroke, which is one of the most common clinical indications of reperfusion injury.

[0276] In another embodiment, the method relates to treating and / or preventing neonatal asphyxia.

[0277] Neonatal asphyxia (or perinatal asphyxia) is a medical condition resulting from a lack of oxygen to a newborn, usually to the brain, during the birth process that lasts long enough to cause physical harm. The most common cause of neonatal asphyxia is a drop in maternal blood pressure during labor or other obstruction to blood flow to the infant's brain, for example, due to inadequate circulation or perfusion, impaired respiratory effort, or inadequate ventilation.

[0278] Neonatal asphyxia can cause hypoxic damage to most of an infant's organs (heart, lungs, liver, gastrointestinal tract, and kidneys), but brain damage is the most severe and is probably the least likely to heal immediately or completely. In more severe cases, the infant survives, but brain damage manifests as either mental disabilities, such as developmental delay or intellectual disability, or physical disabilities, such as spasticity. Infants suffering from severe perinatal asphyxia typically have poor color (cyanosis), poor perfusion, poor responsiveness, poor muscle tone, and inadequate respiratory effort. Excessive asphyxia can lead to cardiac arrest and death. Neonatal asphyxia occurs in 2–10 per 1,000 newborns born at term, with a higher incidence in preterm infants. The WHO estimates that birth asphyxia causes 4 million newborn deaths each year, accounting for 38% of deaths in children under the age of five.

[0279] In another embodiment, the method relates to treating and / or preventing cardiac ischemia, preferably myocardial ischemia.

[0280] In another embodiment, the present invention provides a method for treating and / or preventing stroke, comprising administering to a subject in need thereof: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; simultaneously, sequentially or separately.

[0281] In another embodiment, the present invention provides a method for treating and / or preventing stroke, comprising administering to a subject in need thereof: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. simultaneously, sequentially or separately.

[0282] In another embodiment, the present invention provides a method for treating and / or preventing a neurodegenerative disease, comprising administering to a subject in need thereof: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; simultaneously, sequentially or separately.

[0283] In another embodiment, the present invention provides a method for treating and / or preventing a neurodegenerative disease, comprising administering to a subject in need thereof: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. simultaneously, sequentially or separately.

[0284] In another embodiment, the present invention provides a method of providing neuroprotection comprising administering to a subject in need thereof: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; simultaneously, sequentially or separately.

[0285] In another embodiment, the present invention provides a method of providing neuroprotection comprising administering to a subject in need thereof: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. simultaneously, sequentially or separately.

[0286] In one embodiment, the subject is a mammal, more preferably a human.

[0287] In one embodiment, the method comprises administering the components parenterally (e.g., intravenously, intramuscularly, intradermally, intraperitoneally, or subcutaneously) to a subject. The pharmaceutically active components of the combination can be administered separately or in a combined formulation. Preferably, the pharmaceutically active components are administered separately.

[0288] In another embodiment, the method comprises administering the component to the subject intravenously, intramuscularly, or subcutaneously.

[0289] In another embodiment, the method comprises administering the component to the subject intravenously.

[0290] Each component can be administered by the same or a different route as the other components. Preferably, the components are administered by the same route.

[0291] In one embodiment, the combination is administered to the donor subject and / or recipient subject before, during, and / or after heart transplantation. For example, in some embodiments, the combination can be administered to a first subject from whom a heart organ is removed for transplantation into a second subject. Additionally or alternatively, in some embodiments, the combination is administered to the extracted heart organ before introduction into the second subject. Additionally or alternatively, in some embodiments, the combination therapy is administered to the second subject before, during, and / or after heart transplantation.

[0292] In one embodiment, the combination is for administration to a subject having a stroke. A stroke occurs when insufficient blood flow to the brain leads to cell death. There are two main types of stroke: ischemic, caused by insufficient blood flow, and hemorrhagic, caused by bleeding. As a result, part of the brain stops functioning properly. Signs and symptoms of a stroke include, among others, an inability to move or feel one side of the body, problems understanding or speaking, a feeling that the world is spinning, and loss of vision on one side. Ischemic stroke is usually caused by a blockage in a blood vessel. Treatment for ischemic stroke involves surgery to open (reperfusion) the arteries to the brain in patients with a narrowing problem. If detected within three to four and a half hours, ischemic stroke can sometimes be treated with medications that can destroy blood clots. In 2013, stroke was the second leading cause of death after coronary artery disease, accounting for 6.4 million deaths (12% of all deaths).

[0293] Ischemic stroke and acute myocardial infarction require urgent reperfusion to improve functional outcomes (Patel and Saver, 2013, Stroke,44: 94-98). Intravenous tissue-type plasminogen activator has long been the only reperfusion therapy with proven clinical benefit in patients with acute ischemic stroke. As in acute myocardial infarction, endovascular methods to restore reperfusion in acute ischemic stroke expose patients to increased ischemia / reperfusion injury, which may counteract the benefits of recanalization by promoting hemorrhagic transformation and severe vasogenic edema, both considered markers of reperfusion injury (Bai and Lyden, 2015; Int J Stroke,10: 143-152). Experimental evidence indicates that cerebral ischemic reperfusion injury (as can myocardial reperfusion injury) can be attenuated by ischemic preconditioning and postconditioning. Glibenclamide has been shown to enhance the therapeutic benefits of early hypothermia after severe stroke in rats (Zhu S, et al. Aging Dis. 2018; 9: 685-695). In addition, in a clinically relevant rat model of stroke (middle cerebral artery occlusion), reperfusion was initiated 4.5 hours later, and recombinant tissue plasminogen activator (RTPA) was administered concomitantly. This was followed by glibenclamide (10 μg / kg IP loading dose plus a constant subcutaneous infusion of 200 ng / hour) starting 4.5 or 10 hours after the onset of ischemia; glibenclamide significantly reduced hemispheric swelling at 24 and 48 hours after death and improved the Garcia score at 48 hours, suggesting that the glibenclamide treatment period extends up to 10 hours after the onset of ischemia. This finding is consistent with the observations of a retrospective clinical study suggesting that the use of sulfonylureas is beneficial in the context of rt-PA-assisted recanalization / reperfusion after acute ischemic stroke (Simard, JM, et al. Ann. NY Acad. Sci. 2012;1268:95-107).

[0294] In one embodiment, the combination is intended for administration to a subject suffering from cardiogenic shock. Cardiogenic shock is a life-threatening medical condition resulting from insufficient blood circulation due to primary failure of the heart's ventricles to function effectively. This condition occurs in 2-10% of patients hospitalized for myocardial infarction and is the leading cause of death among these patients (Holmes et al., 1995, J Am Coll Cardiol, 26: 668-674). More specifically, cardiogenic shock is the result of a complex process initiated by cardiac pump failure, involving impaired oxygen delivery, systemic ATP deficiency, and multiple organ dysfunction (Okuda, 2006, Shock, 25: 557-570). Because it is a type of circulatory shock, tissue perfusion is insufficient to meet the demand for oxygen and nutrients. This condition is accompanied by increasingly widespread cell death due to oxygen deprivation (hypoxia) and nutrient deprivation (e.g., hypoglycemia). This condition can lead to cardiac arrest (or circulatory arrest), the sudden cessation of cardiac pumping function (as well as respiratory arrest and loss of consciousness). Cardiogenic shock is defined by sustained hypotension accompanied by tissue hypoperfusion despite adequate left ventricular filling pressure. Signs of tissue hypoperfusion include low urine production (less than 30 mL / hour), cold extremities, and altered level of consciousness. Several large-scale studies have shown that coronary revascularization is the most important strategy for improving patient survival (Hochman et al., 1999, N Engl J Med, 341: 625-634). However, patients who develop cardiogenic shock despite rapid revascularization have a poor prognosis, likely due to reperfusion injury and related to the resulting infarct size. Indeed, hypothermia has been shown to provide tissue protection in myocardial ischemia, and preclinical studies have shown beneficial results in reducing infarct size in experimentally induced myocardial infarction (Dae et al, 2002, Am J Physiol Heart Circ Physiol, 282: H1584-H1591).Thus, in a porcine model, mild therapeutic hypothermia reduced acute mortality and improved hemodynamic parameters in cardiogenic shock (Gotberg et al, 2010, Resuscitation, 81: 1190-96).

[0295] In one embodiment, the combination is for administration to a subject suffering from cardiac arrest. Cardiac arrest is the sudden cessation of effective blood flow due to a failure of the heart's effective contraction. The most common cause of cardiac arrest is coronary artery disease. Treatment for cardiac arrest is immediate cardiopulmonary resuscitation (CPR) and, if a shockable rhythm is present, defibrillation. In the United States, out-of-hospital cardiac arrest occurs in approximately 13 per 10,000 people per year (326,000 cases). In-hospital cardiac arrest occurs in an additional 209,000 people (Kronick et al., Circulation, 2015, 132: S397-S413). In addition to providing high-quality cardiopulmonary resuscitation, optimizing the management of post-resuscitation syndrome is crucial for improving the long-term outcomes of cardiac arrest patients. In this syndrome ("postresuscitation syndrome"), there are three main areas of focus: (1) post-cardiac arrest brain injury; (2) post-cardiac arrest myocardial dysfunction and reperfusion injury; and (3) the systemic ischemia-reperfusion response. It is now clear that post-resuscitation care can affect the long-term survival of survivors as well as myocardial and neurological recovery and function (Kem, 2015, Circ J, 79: 1156-1163).

[0296] In one embodiment, the subject is at risk for (or susceptible to) vaso-occlusive injury or cardiac ischemia-reperfusion injury.

[0297] In one embodiment, the combination is for use in, or a method of, providing neuroprotection in a subject.

[0298] As used herein, the term "neuroprotection" refers to protecting neural entities, including the brain, by preventing, reducing, or delaying brain damage that can lead to neuronal death and neurodegenerative disorders such as Alzheimer's disease, Parkinson's disease, or vascular dementia.

[0299] In one embodiment, the combination is for use in or a method of providing neuroprotection in a subject against the neurotoxic effects of a drug. Examples of neurotoxic drugs are described by Gouzoulis-Mayfrank and Daumann (Dialogues Clin Neurosci. 2009; 11(3): 305-17). Neurotoxic drugs include drugs of abuse (e.g., 3,4-methylenedioxymethamphetamine, methamphetamine, and amphetamine), pesticides (e.g., organophosphate pesticides), certain chemotherapeutics (e.g., platinum), and dopamine.

[0300] In one embodiment, the claimed combination is for use in or a method of providing cardioprotection to a subject against the cardiotoxic effects of a drug (e.g., anthracycline). Examples of cardiotoxic drugs are described in Bovelli et al. (Annals of Oncology 21 (Supplement5): v277-v282, 2010).

[0301] As used herein, the term "cardioprotection" refers to protecting the heart, for example, by preventing, reducing, or delaying myocardial injury. Cardiotoxic drugs include drugs associated with cardiac failure, drugs associated with ischemia or thromboembolism, drugs associated with hypertension, and drugs associated with other toxic effects, such as tamponade and endomyocardial fibrosis, hemorrhagic myocarditis, bradyarrhythmia, Raynaud's phenomenon, autonomic neuropathy, QT prolongation or polymorphic ventricular tachycardia, or pulmonary fibrosis. Examples of cardiotoxic drugs include anthracyclines / anthraquinolones, cyclophosphamide, trastuzumab and other monoclonal antibody-based tyrosine kinase inhibitors, antimetabolites (fluorouracil, capecitabine), microtubule inhibitors (paclitaxel, docetaxel), cisplatin, thalidomide, bevacizumab, sunitinib, sorafenib, busulfan, paclitaxel, vinblastine, bleomycin, vincristine, arsenic trioxide, bleomycin, and methotrexate.

[0302] In one embodiment, the components are administered simultaneously.

[0303] In one embodiment, the components are administered sequentially or separately.

[0304] For combinations of three components, all three components can be administered simultaneously, or any two components can be administered simultaneously and the third component can be administered separately or sequentially, or all three components can be administered separately or sequentially in any order.

[0305] In one embodiment, the sulfonylurea is administered prior to the administration of the insulin modulator, either sequentially or separately.

[0306] In another embodiment, the insulin modulator is administered prior to the administration of the sulfonylurea, either sequentially or separately.

[0307] In one embodiment, the sulfonylurea is administered prior to the administration of the aldosterone antagonist, either sequentially or separately.

[0308] In one embodiment, the aldosterone antagonist is administered prior to the administration of the sulfonylurea, either sequentially or separately.

[0309] In one embodiment, exenatide or a structural or functional analogue or pharmaceutically acceptable salt thereof, canrenoate potassium or a structural or functional analogue thereof, and glibenclamide or a structural or functional analogue thereof are administered sequentially or separately.

[0310] In one embodiment, the components are each administered in a therapeutically effective amount with respect to the individual component.

[0311] As used herein, the term "therapeutically effective amount" refers to an amount sufficient to achieve a desired therapeutic and / or prophylactic effect, e.g., an amount that results in prevention or reduction of ischemia and / or reperfusion injury or one or more symptoms associated with ischemia and / or reperfusion injury.

[0312] In the context of therapeutic or prophylactic use, the amount of the composition administered to a subject depends on the type and severity of the disease, as well as on the individual's characteristics, such as general health, age, sex, weight, and tolerance to drugs. It also depends on the severity and type of the disease. Those skilled in the art can determine the appropriate dosage based on these and other factors. The composition can also be administered in combination with one or more additional therapeutic agents.

[0313] In one embodiment, the components are each administered in a sub-therapeutically effective amount with respect to the individual component.

[0314] In one embodiment, the component is administered prior to reperfusing the subject.

[0315] In one embodiment, the component is administered during reperfusion of the subject.

[0316] In one embodiment, the component is administered after reperfusion of the subject.

[0317] In one embodiment, the component is administered before and / or during and / or after reperfusion of the subject.

[0318] In some embodiments of the method, the subject is administered a sulfonylurea continuously before, during, and after reperfusion of the subject, and an insulin modulator is administered as a bolus dose before reperfusion.

[0319] In some embodiments of the method, the subject is administered an insulin modulator continuously before, during, and after reperfusion of the subject, and a sulfonylurea is administered as a bolus dose before reperfusion.

[0320] In some embodiments of the method, the subject is administered a sulfonylurea continuously before, during, and after reperfusion of the subject, and an aldosterone antagonist is administered as a bolus dose before reperfusion.

[0321] In some embodiments of the method, the subject is administered an aldosterone antagonist continuously before, during, and after reperfusion of the subject, and a sulfonylurea is administered as a bolus dose before reperfusion.

[0322] In some embodiments of the method, the subject is administered the components sequentially before, during, and after reperfusion of the subject.

[0323] In some embodiments of the method, additional administration of one or more components may occur after reperfusion. Preferably, this repeated administration is performed at least twice, more preferably 2 to 100 times, or may be in the form of a continuous infusion.

[0324] In some embodiments of the method, the subject is administered the component as a bolus dose prior to reperfusion.

[0325] In some embodiments of the method, the subject is administered the component as a bolus dose during reperfusion.

[0326] In some embodiments of the method, the subject is administered the component as a bolus dose after reperfusion.

[0327] As used herein, "reperfusion" is the restoration of blood flow to any organ or tissue in which blood flow has been reduced or blocked. For example, blood flow can be restored to any organ or tissue affected by ischemia or hypoxia. Restoration of blood flow (reperfusion) can occur by any method known to those skilled in the art. For example, reperfusion of ischemic cardiac tissue can result from revascularization.

[0328] In one embodiment, reperfusion is achieved by a revascularization procedure selected from the group consisting of percutaneous coronary intervention, balloon angioplasty, insertion of a bypass graft, insertion of a stent, directional coronary atherectomy, treatment with one or more thrombolytic agents, and removal of an obstruction.

[0329] In one embodiment, the one or more thrombolytic agents are selected from the group consisting of tissue plasminogen activator; urokinase; pro-urokinase; streptokinase; acylated forms of plasminogen; acylated forms of plasmin; and acylated streptokinase-plasminogen complex.

[0330] Dosage Those skilled in the art can easily determine the appropriate dosage of one of the compositions to be administered to a subject without undue experimentation.Typically, a doctor will determine the actual dosage that will be most suitable for each patient, and it depends on various factors, including the activity of the specific compound used, the metabolic stability and duration of the action of the compound, age, body weight, general health, sex, diet, mode and time of administration, excretion rate, drug combination, the severity of specific condition and individual ongoing therapy.The dosage disclosed herein is an example of average case.Of course, there may be individual cases where higher or lower dosage ranges are advantageous, and such cases are within the scope of the present invention.

[0331] In one highly preferred embodiment of the present invention, the dose of the insulin modulator (e.g., exenatide) in the combination is generally lower than the dose typically used in monotherapy in the context of its currently approved therapy and / or lower than the common dose reported in the reperfusion injury literature.

[0332] In one highly preferred embodiment of the invention, the dose of the aldosterone antagonist (e.g., potassium canrenoate) in the combination is generally lower than doses typically used in monotherapy in the context of its currently approved therapy and / or lower than common doses reported in the reperfusion injury literature.

[0333] In one highly preferred embodiment of the invention, the dose of the sulfonylurea (e.g., glibenclamide) in the combination is generally lower than the dose typically used in monotherapy in the context of its currently approved therapy and / or lower than the common dose reported in the reperfusion injury literature.

[0334] Each component of the claimed combination may be formulated in unit dosage form, i.e., in the form of discrete portions containing a unit dose, or a multiple or sub-unit of a unit dose. The dosages described herein are applicable to each of the above medical uses.

[0335] When used in the combinations claimed herein, the insulin modulator (e.g., exenatide) is preferably administered at a dose of about 0.001 to about 1.5 μg / kg, more preferably about 0.005 to about 0.15 μg / kg. In one preferred embodiment, the insulin modulator (e.g., exenatide) is preferably administered at a dose of about 0.01 to about 1.5 μg / kg, more preferably about 0.05 to about 1.5 μg / kg. As used herein, the dosage of the insulin modulator is in μg / kg body weight (μg = microgram).

[0336] In one preferred embodiment, the insulin modulator (e.g., exenatide) is administered at a dose of preferably about 0.01 to about 0.5 μg / kg, more preferably about 0.02 to about 0.5 μg / kg, or about 0.03 to about 0.5 μg / kg, or about 0.04 to about 0.5 μg / kg, or about 0.05 to about 0.5 μg / kg, or about 0.05 to about 0.2 μg / kg, or about 0.05 to about 0.15 μg / kg.

[0337] In one preferred embodiment, the insulin modulator (e.g., exenatide) is administered at a dose of preferably about 0.01 to about 0.1 μg / kg, more preferably about 0.02 to about 0.08 μg / kg, or about 0.03 to about 0.07 μg / kg, or about 0.04 to about 0.06 μg / kg, or at a dose of about 0.05 μg / kg.

[0338] When used in the combinations claimed herein, the aldosterone antagonist (e.g., potassium canrenoate) is preferably administered at a dose of about 0.03 to about 10 mg / kg, or about 0.1 to about 10 mg / kg, or about 0.3 to about 5 mg / kg, or about 1 to about 10 mg / kg, or about 1 to about 5 mg / kg, or about 1 to about 3 mg / kg. As used herein, the dose of the aldosterone antagonist is in mg / kg body weight.

[0339] In one preferred embodiment, the aldosterone antagonist (e.g., potassium canrenoate) is preferably administered at a dose of about 0.1 to about 3 mg / kg, or about 0.2 to about 2 mg / kg, or about 0.3 to about 1.5 mg / kg, or about 0.3 to about 1 mg / kg.

[0340] In one preferred embodiment, the aldosterone antagonist (e.g., potassium canrenoate) is administered at a dose of about 0.1 to about 0.5 mg / kg, or about 0.2 to about 0.5 mg / kg, more preferably about 0.2 to about 0.4 mg / kg, and even more preferably about 0.3 to about 0.4 mg / kg.

[0341] When used in the combinations claimed herein, the sulfonylurea (e.g., glibenclamide) is preferably administered at a dose of about 0.001 to about 30 μg / kg, more preferably about 0.01 to about 5 μg / kg, and even more preferably about 0.01 to about 2 μg / kg. As used herein, sulfonylurea dosages are in μg / kg body weight.

[0342] In one preferred embodiment, the sulfonylurea (e.g., glibenclamide) is preferably administered at a dose of about 0.5 to about 20 μg / kg, or about 0.5 to about 15 μg / kg, or about 0.5 to about 10 μg / kg, or about 1 to about 10 μg / kg.

[0343] In one preferred embodiment, the sulfonylurea (e.g., glibenclamide) is administered at a dose of about 0.5 to about 8 μg / kg, or about 0.5 to about 7 μg / kg, or about 0.5 to about 6 μg / kg, or about 0.5 to about 5 μg / kg. In one preferred embodiment, the sulfonylurea (e.g., glibenclamide) is administered at a dose of about 0.5 to about 3 μg / kg, or about 0.5 to about 2 μg / kg, or about 0.5 to about 1.5 μg / kg, or about 0.8 to about 1.2 μg / kg, or about 1 μg / kg.

[0344] In one highly preferred embodiment, the combination is a fixed-dose combination comprising a predetermined dosage of each pharmaceutically active ingredient, thereby allowing the subject to be administered the dosages described above, e.g., about 0.005 to about 0.15 μg / kg of exenatide and about 0.001 to about 30 μg / kg of glibenclamide.

[0345] Preferably, the fixed dose combination comprises predetermined doses of each active pharmaceutical ingredient to enable the following doses to be administered to a subject:

[0346] In one highly preferred embodiment, the combination is a fixed dose combination comprising about 0.01 to about 0.5 μg / kg of exenatide and about 0.5 to about 20 μg / kg of glibenclamide.

[0347] In one highly preferred embodiment, the combination is a fixed dose combination comprising about 0.01 to about 0.1 μg / kg of exenatide and about 0.5 to about 8 μg / kg of glibenclamide.

[0348] In one highly preferred embodiment, the combination is a fixed dose combination comprising about 0.01 to about 0.1 μg / kg of exenatide and about 0.5 to about 1.5 μg / kg of glibenclamide.

[0349] In one highly preferred embodiment, the combination is a fixed-dose combination comprising a predetermined dosage of each component, for example, from about 0.03 to about 10 mg / kg of potassium canrenoate and from about 0.001 to about 30 μg / kg of glibenclamide.

[0350] In one highly preferred embodiment, the combination is a fixed dose combination comprising about 0.1 to about 3 mg / kg of potassium canrenoate and about 0.5 to about 20 μg / kg of glibenclamide.

[0351] In one highly preferred embodiment, the combination is a fixed dose combination comprising about 0.1 to about 0.5 mg / kg of potassium canrenoate and about 0.5 to about 8 μg / kg of glibenclamide.

[0352] In one highly preferred embodiment, the combination is a fixed dose combination comprising about 0.1 to about 0.5 mg / kg of potassium canrenoate and about 0.5 to about 1.5 μg / kg of glibenclamide.

[0353] In one highly preferred embodiment, the combination is a fixed dose combination comprising a predetermined dosage of each component, for example, about 0.03 to about 10 mg / kg of potassium canrenoate, about 0.001 to about 30 μg / kg of glibenclamide, and about 0.005 to about 0.15 μg / kg of exenatide.

[0354] In one highly preferred embodiment, the combination is a fixed dose combination comprising about 0.1 to about 3 mg / kg of potassium canrenoate, about 0.5 to about 20 μg / kg of glibenclamide, and about 0.01 to about 0.5 μg / kg of exenatide.

[0355] In one highly preferred embodiment, the combination is a fixed dose combination comprising about 0.1 to about 0.5 mg / kg of potassium canrenoate, about 0.5 to about 8 μg / kg of glibenclamide, and about 0.01 to about 0.1 μg / kg of exenatide.

[0356] In one highly preferred embodiment, the combination is a fixed dose combination comprising about 0.1 to about 0.5 mg / kg of potassium canrenoate, about 0.5 to about 1.5 μg / kg of glibenclamide, and about 0.01 to about 0.1 μg / kg of exenatide.

[0357] In one highly preferred embodiment, the combination is a fixed dose combination comprising about 0.05 μg / kg of exenatide and 1 μg / kg of glibenclamide.

[0358] In one highly preferred embodiment, the combination is a fixed dose combination comprising about 0.33 mg / kg of potassium canrenoate and about 1 μg / kg of glibenclamide.

[0359] In one highly preferred embodiment, the combination is a fixed dose combination comprising about 0.05 μg / kg of exenatide, about 0.33 mg / kg of potassium canrenoate, and about 1 μg / kg of glibenclamide.

[0360] Non-therapeutic use In another aspect, the present invention provides a method for treating and / or preventing ischemia and / or reperfusion injury in an ex vivo organ prior to or during transplantation, comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to the use of a combination comprising:

[0361] In another preferred embodiment, the present invention provides a method for treating and / or preventing ischemia and / or reperfusion injury in an ex vivo organ before or during transplantation, comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. The present invention relates to the use of a combination comprising:

[0362] In one embodiment, the present invention provides a method for treating and / or preventing reperfusion injury in an ex vivo organ prior to or during transplantation, comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to the use of a combination comprising:

[0363] In one preferred embodiment, the present invention provides a method for treating and / or preventing reperfusion injury in an ex vivo organ prior to or during transplantation, comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; (ii) canrenoate potassium, or a structural or functional analog thereof; The present invention relates to the use of a combination comprising:

[0364] In one embodiment, the present invention provides a method for treating and / or preventing ischemia in an ex vivo organ prior to or during transplantation, comprising: (a) with sulfonylureas; (b) at least one of the following components: (i) an insulin modulator, and (ii) an aldosterone antagonist; The present invention relates to the use of a combination comprising:

[0365] In one preferred embodiment, the present invention provides a method for treating and / or preventing ischemia in an ex vivo organ prior to or during transplantation, comprising: (a) glibenclamide, or a structural or functional analog thereof; (b) at least one of the following ingredients: (i) exenatide, or a structural or functional analog thereof, or a pharmaceutically acceptable salt thereof; and (ii) canrenoate potassium, or a structural or functional analog thereof. The present invention relates to the use of a combination comprising:

[0366] Ex vivo (removed from the body) organs may be susceptible to reperfusion injury due to lack of blood flow. Therefore, the combination of the present invention can be used to prevent reperfusion injury in removed organs. Preferably, the organ is the heart, liver, or kidney, more preferably the heart.

[0367] In some embodiments, the removed organ is placed in a standard buffer solution, such as a solution commonly used in the art, containing the combination of the present invention. For example, the removed heart can be placed in a cardioplegic solution containing exenatide, potassium canrenoate, and glibenclamide. The concentrations of exenatide, potassium canrenoate, and glibenclamide useful in standard buffer solutions can be easily determined by those skilled in the art. Such concentrations may be, for example, about 0.1 nM to about 10 μM, preferably about 1 nM to about 10 μM.

[0368] The present invention will now be further described with reference to the accompanying non-limiting examples and the following figures: [Brief explanation of the drawings]

[0369] [Figure 1] Figure 1 shows the relative cerebral infarct volume (%) at day 7 after repeated administration (for 7 days) of a low dose of the triple combination (Treatment S) versus the relative cerebral infarct volume (%) at day 7 in control animals in Part I and Part II, and in animals receiving the corresponding monotherapies (Treatments A, E, and I) and the corresponding double combinations (Treatments M, N, and O) in a rat model of transient middle cerebral artery occlusion. Treatment A: exenatide 0.05 μg / kg (n=4); Treatment E: canrenoate potassium 0.33 mg / kg (n=3); Treatment I: glibenclamide 1 μg / kg (n=4); Treatment M: exenatide 0.05 μg / kg and canrenoate potassium 0.33 mg / kg (n=13); Treatment N: exenatide 0.05 μg / kg and glibenclamide 1 μg / kg (n=12); Treatment O: canrenoate potassium 0.33 mg / kg and glibenclamide 1 μg / kg (n=11); Treatment S: exenatide 0.05 μg / kg and canrenoate potassium 0.33 mg / kg and glibenclamide 1 μg / kg (n=7); Control Part I (n=6); Control Part II (n=5). [Figure 2]FIG. 1 shows the modified neurological severity scores on day 2 after repeated administration (7 days) of a low dose of the triple combination (Treatment S) versus the modified neurological severity scores on day 2 in control animals in Part I and Part II, and in animals receiving the corresponding monotherapies (Treatments A, E, and I) and the corresponding dual combinations (Treatments M, N, and O) in a rat model of transient middle cerebral artery occlusion. Treatment A: exenatide 0.05 μg / kg (n=4); Treatment E: canrenoate potassium 0.33 mg / kg (n=3); Treatment I: glibenclamide 1 μg / kg (n=4); Treatment M: exenatide 0.05 μg / kg and canrenoate potassium 0.33 mg / kg (n=13); Treatment N: exenatide 0.05 μg / kg and glibenclamide 1 μg / kg (n=12); Treatment O: canrenoate potassium 0.33 mg / kg and glibenclamide 1 μg / kg (n=11); Treatment S: exenatide 0.05 μg / kg and canrenoate potassium 0.33 mg / kg and glibenclamide 1 μg / kg (n=7); Control Part I (n=6); Control Part II (n=5). [Figure 3] FIG. 1 shows the modified neurological severity scores at day 7 after repeated administration (for 7 days) of a low dose of the triple combination (Treatment S) versus the modified neurological severity scores at day 7 in control animals in Part I and Part II, and in animals receiving the corresponding monotherapies (Treatments A, E, and I) and the corresponding dual combinations (Treatments M, N, and O) in a rat model of transient middle cerebral artery occlusion. Treatment A: exenatide 0.05 μg / kg (n=4); Treatment E: canrenoate potassium 0.33 mg / kg (n=3); Treatment I: glibenclamide 1 μg / kg (n=4); Treatment M: exenatide 0.05 μg / kg and canrenoate potassium 0.33 mg / kg (n=13); Treatment N: exenatide 0.05 μg / kg and glibenclamide 1 μg / kg (n=12); Treatment O: canrenoate potassium 0.33 mg / kg and glibenclamide 1 μg / kg (n=11); Treatment S: exenatide 0.05 μg / kg and canrenoate potassium 0.33 mg / kg and glibenclamide 1 μg / kg (n=7); Control Part I (n=6); Control Part II (n=5). [Figure 4]Figure 1 shows the relative cerebral infarct volume (%) at day 7 after repeated administration (for 7 days) of a high dose of the triple combination (Treatment T) versus the relative cerebral infarct volume (%) at day 7 in control animals in Part I and Part II, and in animals receiving the corresponding monotherapies (Treatments B, F, and K) and the corresponding double combinations (Treatments Q and R) in a rat model of transient middle cerebral artery occlusion. Treatment B: exenatide 0.15 μg / kg (n=8); Treatment F: canrenoate potassium 1 mg / kg (n=8); Treatment K: glibenclamide 10 μg / kg (n=8); Treatment Q: exenatide 0.15 μg / kg and glibenclamide 10 μg / kg (n=4); Treatment R: canrenoate potassium 1 mg / kg and glibenclamide 10 μg / kg (n=5); Treatment T: exenatide 0.15 μg / kg, canrenoate potassium 1 mg / kg, and glibenclamide 10 μg / kg (n=7). Control Part I (n=6); Control Part II (n=5). [Figure 5] FIG. 1 shows the modified neurological severity scores on day 2 after repeated administration (7 days) of a high dose of the triple combination (Treatment T) versus the modified neurological severity scores on day 2 in control animals in Part I and Part II, and in animals receiving the corresponding monotherapies (Treatments B, F, and K) and the corresponding dual combinations (Treatments Q and R) in a rat model of transient middle cerebral artery occlusion. Treatment B: exenatide 0.15 μg / kg (n=8); Treatment F: canrenoate potassium 1 mg / kg (n=8); Treatment K: glibenclamide 10 μg / kg (n=8); Treatment Q: exenatide 0.15 μg / kg and glibenclamide 10 μg / kg (n=4); Treatment R: canrenoate potassium 1 mg / kg and glibenclamide 10 μg / kg (n=5); Treatment T: exenatide 0.15 μg / kg, canrenoate potassium 1 mg / kg, and glibenclamide 10 μg / kg (n=7). Control Part I (n=6); Control Part II (n=5). [Figure 6]FIG. 1 shows the modified neurological severity scores at day 7 after repeated administration (for 7 days) of a high dose of the triple combination (Treatment T) versus the modified neurological severity scores at day 7 in control animals in Part I and Part II, and in animals receiving the corresponding monotherapies (Treatments B, F, and K) and the corresponding dual combinations (Treatments Q and R) in a rat model of transient middle cerebral artery occlusion. Treatment B: exenatide 0.15 μg / kg (n=8); Treatment F: canrenoate potassium 1 mg / kg (n=8); Treatment K: glibenclamide 10 μg / kg (n=8); Treatment Q: exenatide 0.15 μg / kg and glibenclamide 10 μg / kg (n=4); Treatment R: canrenoate potassium 1 mg / kg and glibenclamide 10 μg / kg (n=5); Treatment T: exenatide 0.15 μg / kg, canrenoate potassium 1 mg / kg, and glibenclamide 10 μg / kg (n=7). Control Part I (n=6); Control Part II (n=5). [Figure 7] Figure 7 shows the results of histological TUNEL staining of apoptosis in the hippocampus region of rats treated with the triple combination of exenatide / canrenoate potassium / glibenclamide in a rat model of vascular dementia. More specifically, Figure 7 shows the percentage (mean ± SEM) of apoptotic cells in rats treated with exenatide 0.05 μg / kg + canrenoate potassium 0.33 mg / kg + glibenclamide 1 μg / kg (group 2M; 13 animals; intravenous administration) compared to vehicle-treated controls (group 1M; 9 animals). [Example]

[0370] This invention is further illustrated by the following examples, which should not be construed as limiting in any way. Example 1

[0371] Dose-response study of the efficacy of exenatide, canrenoate potassium, and glibenclamide and their combination in a rat model of cerebral ischemia and reperfusion injury The objectives of this study were to evaluate (a) the dose-response of the neuroprotective effects of glibenclamide, exenatide, and potassium canrenoate in a rat model of cerebral ischemia and reperfusion injury after repeated intravenous administration as monotherapy (Study Part I) and (b) the effects of the compound combination versus the corresponding monotherapy (Study Part II).

[0372] Transient middle cerebral artery occlusion (t-MCAO) was performed according to the method described by R. Schmid-Elsaesser et al. (Stroke. 1998; 29(10): 2162-70). Test compounds were administered intravenously 20 minutes before reperfusion and then twice daily for 6 consecutive days. The modified neurological severity score (NSS) was graded on a scale of 0 to 18 (with a normal score of 0 and a maximum deficit score of 18) on study day 2 (the day after surgery) and study day 7 (7 days after surgery and before study termination); this included a series of clinical neurological examinations (a composite of motor, sensory, reflex, and balance tests). At the end of the study, brains were harvested, sliced ​​into five 2-mm-thick coronal sections, and stained with triphenyl tetrazolium chloride (TTC). Infarct size was measured using the ImageJ program. Morbidity, mortality, body weight, and clinical observations were also recorded. Numerical results are presented as the mean ± standard deviation of the mean. Statistical significance (P) of treated versus untreated controls was determined using two-way ANOVA followed by Bonferroni post-hoc tests using the GraphPad Prism5 program.

[0373] In Part I of the study, a total of 112 male SD rats (270-320 gr on arrival) were divided into 13 groups (5 or 10 rats per treatment group and 8 rats in the control group). Due to mortality in some groups, numbers within groups were adjusted to ensure there were enough animals in all groups. The groups were as follows: Control (saline) Exenatide administered at 0.05 μg / kg, 0.15 μg / kg, 0.5 μg / kg, and 1.5 μg / kg; canrenoate potassium administered at 0.33 mg / kg, 1 mg / kg, 3 mg / kg, and 10 mg / kg; Glibenclamide administered at 1 μg / kg, 3 μg / kg, 10 μg / kg and 30 μg / kg.

[0374] The efficacy endpoint results obtained in Part I are summarized in Table 1, which also includes the results of statistical comparisons of each treatment group versus the matched control group, expressed as a percentage change relative to the matched control. Twenty-seven animals died during the study across all groups (1 during surgery, 5 after occlusion, 1 euthanized on day 2, 7 immediately after reperfusion, and 13 found dead in their cages within 1 to 5 days after surgery). There were no statistically significant differences in body weight between all animal groups.

[0375] With the exception of the lowest doses of exenatide, canrenoate potassium, and glibenclamide (Treatments A, E, and I), all monotherapies demonstrated statistically significant reductions in cerebral infarct size when compared with the control group. There was no clear indication of a dose response.

[0376] For modified neurological severity score (NSS), only the lowest doses of exenatide, canrenoate potassium, and glibenclamide (Treatments A, E, and I) and the 3 μg / kg dose of glibenclamide (Treatment J) showed no statistically significant reduction compared to the control group on day 2, whereas only the lowest doses of exenatide, canrenoate potassium, and glibenclamide (Treatment A, E, and I) and the 0.5 μg / kg dose of exenatide (Treatment C) showed no statistically significant reduction compared to the control group on day 7. As with cerebral infarct size, there was no clear indication of a dose response for modified NSS on any day.

[0377] For Part II of the study, a total of 86 SD male rats (270-320 gr on arrival) were divided into 9 groups (7 or 15 rats per treatment group, 6 rats in the control group). Due to mortality in some groups, numbers within groups were adjusted to ensure there were enough animals in all groups. The groups were as follows: Control (saline) exenatide administered at 0.05 μg / kg and potassium canrenoate administered at 0.33 mg / kg; exenatide administered at 0.05 μg / kg and glibenclamide administered at 1 μg / kg; canrenoate potassium administered at 0.33 mg / kg and glibenclamide administered at 1 μg / kg; exenatide administered at 0.15 μg / kg and potassium canrenoate administered at 0.33 mg / kg; exenatide administered at 0.15 μg / kg and glibenclamide administered at 10 μg / kg; potassium canrenoate administered at 1 mg / kg and glibenclamide administered at 10 μg / kg; exenatide administered at 0.05 μg / kg, and potassium canrenoate administered at 0.33 mg / kg and glibenclamide administered at 1 μg / kg; Exenatide administered at 0.15 μg / kg, and potassium canrenoate administered at 1 mg / kg and glibenclamide administered at 10 μg / kg.

[0378] The efficacy endpoint results obtained in Part II are summarized in Table 2, which also includes the results of statistical comparisons of each treatment group versus the matched control group, expressed as a percentage change relative to the matched control. Nineteen animals died during the study across all groups (two were euthanized on day 6, four died immediately after reperfusion, and 13 were found dead in their cages within one to five days after surgery). There were no statistically significant differences in body weights among all animal groups.

[0379] All dual combinations (Groups M, N, O, P, Q, and R) and both triple combinations (Groups S and T) showed statistically significant changes in cerebral infarct size when compared with the control group. There were no differences between dual combinations (Groups M, N, O, P, Q, and R) or triple combinations (Groups S and T). The reduction in cerebral infarct size after the dual combination of the lowest dose of exenatide and potassium canrenoate (Group M) was statistically different from that of the corresponding monotherapy (Groups A and E). The reduction in cerebral infarct size after the dual combination of the lowest dose of exenatide and glibenclamide (Group N) was statistically different only from that of the corresponding glibenclamide monotherapy (Group I). Furthermore, the low-dose triple combination (Group S), but not the high-dose triple combination (Group T), demonstrated statistically significant reductions in cerebral infarct size compared with the corresponding monotherapy.

[0380] All double combinations (Groups M, N, O, P, Q, and R) and both triple combinations (Groups S and T) demonstrated statistically significant reductions in modified neurological severity scores (NSS) when compared with the control group on Days 2 and 7. Furthermore, the reduction in modified NSS with the triple combination of the lowest dose of exenatide, canrenoate potassium, and glibenclamide (Group S) was statistically significantly different from the corresponding canrenoate potassium and glibenclamide monotherapies (Groups E and I) on Day 2 and from all three corresponding monotherapies (Groups A, E, and I) on Day 7, but not from any of the corresponding double combinations (Groups M, N, and O). The triple combination of high-dose exenatide, canrenoate potassium, and glibenclamide (Group T) was not statistically significantly different from any of the corresponding double combinations (Groups Q and R).

[0381] To assess the efficacy of the combination therapy, the lowest dose triple combination (Treatment "S") was compared with the corresponding monotherapy and double combination for relative cerebral infarct volume and modified neurological severity score at days 2 and 7, respectively, as shown in Figures 1-3. Similarly, the results of the comparison of the high dose triple combination (Treatment "T") versus the corresponding monotherapy and double combination are shown in Figures 4-6.

[0382] The results demonstrate that low doses of exenatide, canrenoate potassium, and glibenclamide, which are ineffective when administered as monotherapy, demonstrate statistically significant efficacy and synergy when combined (both as double or triple combinations). The results support the combined effects of glibenclamide with exenatide and / or canrenoate potassium, as the combined effects achieved exceeded the sum of the effects of each monotherapy. Reduce the size of the stroke, and / or Improve neurological severity scores, and / or This study provides strong evidence that glibenclamide synergistically improves motor performance scores. Furthermore, a surprising finding is that the dose of glibenclamide that produced this synergistic effect in this study (i.e., 1 μg / kg twice daily, or 0.66 μg for the 330 g rats used in the study) was significantly lower than the dose previously reported in the stroke literature, i.e., a daily infusion of 200 ng / hour, or 4.8 μg (Simard et al., Transl Stroke Res. 2012). Importantly, such a very low dose of glibenclamide when used in the context of this invention corresponds to a dose (i.e., 70 μg / day) that is less than 1 / 100th of the prescribed daily dose (7 mg oral micronized formulation) or approximately 20-285 times less than the recommended maintenance dose of glibenclamide (micronized formulation), and therefore is not expected to affect blood glucose levels or have any adverse effects. The above clinically effective doses of glibenclamide in double or triple combinations with low-dose exenatide and / or potassium carbonate have also been shown to be significantly lower than the doses of glibenclamide shown to be neuroprotective in published clinical studies (0.16 or 0.11 mg / hour continuous infusion, i.e., 3.84 mg or 2.64 mg per day) (see King ZA, et al.). Example 2

[0383] Study of the efficacy of a combination of exenatide, canrenoate potassium, and glibenclamide in a rat model of vascular dementia The chronic cerebral hypoperfusion model in Wistar rats involves permanent occlusion of both common carotid arteries, resulting in brain lesions that may also affect cognitive function. This model is similar to models of vascular dementia, and this technique can reduce blood flow to the cerebral cortex and hippocampus by up to 40-80% for several months, resulting in certain learning disabilities.

[0384] Research purpose The aim of this study was to evaluate the neuroprotective effect of a combination of exenatide, canrenoate potassium, and glibenclamide given intravenously, twice daily for 3 weeks, 24 hours after permanent ligation of both common carotid arteries, using a Wistar rat model of vascular dementia.

[0385] treatment group The treatment groups were as follows: Group 1M: vehicle-treated control (9 animals, administered intravenously); Group 2M: exenatide 0.05 μg / kg + potassium canrenoate 0.33 mg / kg + glibenclamide 1 μg / kg (13 animals; administered intravenously).

[0386] Exenatide acetate was obtained from Bachem AG, Switzerland. Canrenoate potassium was obtained from Pfizer, Switzerland. Glibenclamide was obtained from Tocris Bioscience.

[0387] Study Design and Timeline This study evaluated the neuroprotective effects of the combination administered intravenously at low doses twice daily for three weeks in a Wistar rat model of vascular dementia. The test compounds were administered twice daily for three weeks, 24 hours after common carotid artery ligation. On day 1, both common carotid arteries were permanently ligated. Morris water maze testing was performed before common carotid artery ligation as baseline training, and at weeks 4 and 8 thereafter. Brains were harvested at the end of the study. Histological analysis of tissues was performed.

[0388] The timeline of the study is as follows:

[0389] [Table 1]

[0390] The first administration date was assigned as "Day 1," and the end date was "Day 56," 8 weeks after common carotid artery ligation.

[0391] Histology analysis Tissue preparation and trimming (affected hemisphere), precise sectioning of the striatum (corpus callosum), and dorsal hippocampus x3, and optical tracks per brain. Paraffin block preparation, H&E and TUNEL staining, IHC: doublecortin for neuronal regeneration in the subventricular zone of the brain, MBP, myelin in the white matter, Iba-1 for microglia, and GFAP for astrocytes, Olig-2 for all oligodendrocytes, and NG2 for young oligodendrocytes. Slide evaluation analysis: cell body counting in the CA1 and CA3 regions of the hippocampus - 3 sections per brain, neuronal death count in 3 fields per section, and morphometric analysis of MBP.

[0392] animal Male Wistar rats were used in the study, weighing 290-390 g at the start of the study.

[0393] animal control breeding Animal care was conducted in accordance with the guidelines of the National Institutes of Health (NIH) and the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). Animals were housed in 42.5 × 26.5 × 18.5 cm polyethylene cages (maximum of three rats / cage) equipped with stainless steel grill-tops to facilitate access to pelleted food and bottled drinking water; bedding: steam-sterilized clean rice husks (Envigo, Sani-chips catalog number 7090C). Bedding was changed along with the cages at least twice weekly.

[0394] feed Animals were fed a commercial rodent diet (Teklad Certified Global 18% Protein Diet, Envigo, Cat. No. 2018SC) ad libitum. Animals had ad libitum access to standard tap drinking water obtained from a municipal source and treated according to Pharmaseed's SOP No. 214: "Water system." Animal diets arrived with a certificate of analysis, and water was autoclaved before use.

[0395] environmental conditions Animals were housed singly for the postoperative period in a temperature-controlled environment. The air was filtered (HEPA F6 / 6) with an adequate fresh supply (minimum 15 air changes / hour). The temperature was maintained between 18 and 24°C and the relative humidity between 30 and 70%. Animals were exposed to a 12-hour light and 12-hour dark cycle (6 AM / 6 PM).

[0396] Randomization Animals were randomly assigned to cages according to Pharmaseed's SOP#027 "Random allocation of animals."

[0397] Treatment and Evaluation Surgical procedure: ligation of two common carotid arteries On the day of surgery, anesthesia was induced with 4% isoflurane in a mixture of 70% NO and 30% O on a heating pad and maintained with 1.5–2% isoflurane. 0.1 mg / kg buprenorphine was injected subcutaneously. Two common carotid artery occlusions were performed according to the method described by Hyun Joon Lee et al. (Citicoline Protects Against Cognitive Impairment in a Rat Model of Chronic Cerebral Hypoperfusion, J Clin Neurol. 2009; 5(1):33-38). Both common carotid arteries (CCA) were exposed through a midline neck incision and carefully dissected free of surrounding nerves and fascia. Both arteries were double-ligated with 4-0 silk sutures 8–10 mm below the visible area of ​​the external carotid artery. The surgical wound was closed, and the animal was returned to its cage and allowed to recover from anesthesia. Analgesic treatment with buprenorphine was resumed by the end of the day and twice daily for the next 4 days.

[0398] Combination Administration Treatment was initiated 24 hours after arterial ligation by intravenous (IV) injection twice daily for three consecutive weeks.

[0399] body weight Animal weights were monitored during acclimation, before common carotid artery ligation, and twice weekly thereafter. Animals were weighed according to Pharmaseed's SOP No. 010: "Weighing laboratory animals." Individual weight changes were calculated.

[0400] Clinical observations Clinical signs were monitored once during acclimatization, during the first 4 hours postoperatively, twice daily for the first 2 days postoperatively, and twice weekly thereafter.

[0401] Morris water maze test The Morris water maze (MWM) test is designed to assess cognitive impairment after common carotid artery ligation. The test was performed according to Pharmaseed's SOP 100 (Morris Water Maze Testing V6) and related publications (e.g., Brandeis R, Brandys Y and Yehuda S, "The use of the Morris Water Maze in the study of memory and learning", Int J Neurosci. 1989; 48(1-2):29-69).

[0402] Pre-operative training Animals were trained and conditioned in the Morris water maze for 1 week according to Pharmaseed's SOP100 and scientific publications (see, e.g., Brandeis R et al.). Prior to the MWM, rat cages were moved from the animal house to the behavioral testing room and allowed to acclimate for approximately 1 hour.

[0403] The training results on the final day were considered as baseline data for comparison.,The MWM test had the following exclusion criteria: failure to,escape to the platform within 90 seconds (the third day of training).

[0404] After common carotid artery ligation test Before the MWM, rat cages were moved from the animal house to the behavioral testing room and allowed to acclimate for approximately 1 hour.

[0405] The MWM test was performed 4 and 8 weeks after common carotid artery ligation.

[0406] statistical analysis Numerical results are presented as means and standard deviations or standard errors. Descriptive statistics of data and group comparisons, whenever possible, were performed using a statistical analysis program (GraphPad Prism version 5.02 for Windows, GraphPad Software, San Diego, California, USA). Appropriate parametric or nonparametric tests were performed, followed by appropriate post-hoc analyses. A probability of 5% (p≦0.05) was considered statistically significant.

[0407] result Preliminary results of the MWM test indicate that animals treated with the exenatide / potassium / glibenclamide triple combination (treatment group 2M; 13 animals) performed better than vehicle-treated animals (group 1M; 9 animals) in terms of both the mean time (seconds) to reach the platform and the mean distance (cm) swam to the platform. Furthermore, quantitative assessment of histological TUNEL staining for apoptosis in the hippocampal region demonstrated statistically significant neuroprotection (p = 0.03 by Mann-Whitney test) for group 2M treatment compared with group 1M (vehicle). Figure 7 shows the percentage of apoptotic cells (mean ± SEM) for each group.

[0408] Therefore, it can be concluded that the triple combination of exenatide / potassium / glibenclamide, when administered intravenously at low doses twice daily for 3 weeks, exhibited neuroprotective effects in the Wistar rat model of vascular dementia, showing both improved cognitive behavior and reduced apoptosis in the hippocampal brain region.

[0409] Various modifications and variations of the described aspects of the invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention that are obvious to those skilled in the relevant fields are intended to be within the scope of the following claims.

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[0411] [Table 2]

[0412] [Table 3]

Claims

1. (a) a sulfonylurea that is glibenclamide; (b) an aldosterone antagonist that is canrenoate potassium; and (c) an insulin modulator that is exenatide or a pharmaceutically acceptable salt thereof, or a structural or functional analog of exenatide selected from lixisenatide, albiglutide, liraglutide, taspoglutide, and dulaglutide (LY2189265); and A combination including:

2. 10. The combination of claim 1, comprising at least one further active pharmaceutical ingredient (API) selected from beta blockers, renin-angiotensin inhibitors, statins (HMG-CoA reductase inhibitors), inhibitors of platelet activation or aggregation, phosphodiesterase-3 inhibitors, calcium sensitizers, antioxidants, and anti-inflammatory agents.

3. 3. A pharmaceutical composition comprising a combination according to claim 1 or 2 and a pharmaceutically acceptable carrier, diluent or excipient.

4. 4. The pharmaceutical composition of claim 3 in a form suitable for parenteral administration.

5. 5. The pharmaceutical composition of claim 4 in a form suitable for intravenous administration.

6. (a) a sulfonylurea that is glibenclamide; (b) an aldosterone antagonist that is canrenoate potassium; and (c) an insulin modulator that is exenatide or a pharmaceutically acceptable salt thereof, or a structural or functional analog of exenatide selected from lixisenatide, albiglutide, liraglutide, taspoglutide, and dulaglutide (LY2189265); and Pharmaceutical products including:

7. 10. A combination according to claim 1 or 2, or a pharmaceutical composition according to claim 4 or 5, or a pharmaceutical product according to claim 6, for use in the treatment and / or prevention of one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative diseases, neonatal asphyxia, cardiac arrest, cardiogenic shock and acute myocardial infarction, or for use in providing cardioprotection against cardiotoxic drugs, or for use in providing neuroprotection.

8. 4. The pharmaceutical composition of claim 3 for use in the treatment and / or prevention of one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for use in providing cardioprotection against cardiotoxic drugs, or for use in providing neuroprotection.

9. 8. The combination or pharmaceutical composition or pharmaceutical product for use according to claim 7, wherein the ischemia and / or reperfusion injury is cerebral, cardiac, pulmonary, or renal ischemia and / or reperfusion injury.

10. 9. The combination or pharmaceutical composition or pharmaceutical product for use according to claim 8, wherein the ischemia and / or reperfusion injury is cerebral, cardiac, pulmonary, or renal ischemia and / or reperfusion injury.

11. 11. The combination or pharmaceutical composition or pharmaceutical product for use according to claim 9 or 10, wherein the ischemia and / or reperfusion injury is cerebral ischemia, cerebral reperfusion injury or stroke.

12. The combination or pharmaceutical composition or pharmaceutical product for use according to claim 7, wherein (a) to (c) are for intravenous administration.

13. The combination or pharmaceutical composition or pharmaceutical product for use according to claim 8, wherein (a) to (c) are for intravenous administration.

14. 8. The combination or pharmaceutical composition or pharmaceutical product for use according to claim 7, wherein (a) to (c) are for administration during, before or after reperfusion.

15. 9. The combination or pharmaceutical composition or pharmaceutical product for use according to claim 8, wherein (a) to (c) are for administration during, before or after reperfusion.

16. 7. The pharmaceutical product of claim 6, for use in the treatment and / or prevention of one or more of the following: ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for use in providing cardioprotection against cardiotoxic drugs, or for use in providing neuroprotection, wherein (a) to (c) are for simultaneous, sequential, or separate administration.

17. 17. The pharmaceutical product for use according to claim 16, wherein (a) to (c) are for parenteral administration.

18. 18. The pharmaceutical product for use according to claim 17, wherein (a) to (c) are for intravenous administration.

19. The pharmaceutical product for use according to any one of claims 16 to 18, wherein (a) to (c) are for simultaneous administration.

20. In the manufacture of a medicament for treating and / or preventing one or more of ischemia and / or reperfusion injury, stroke, neurodegenerative disease, neonatal asphyxia, cardiac arrest, cardiogenic shock, and acute myocardial infarction, or for providing cardioprotection against cardiotoxic drugs, or for providing neuroprotection, (a) glibenclamide; and (b) (i) exenatide, or a pharmaceutically acceptable salt thereof, or a structural or functional analog of exenatide selected from lixisenatide, albiglutide, liraglutide, taspoglutide, and dulaglutide (LY2189265); and (ii) canrenoate potassium; Use of.

21. 21. The use according to claim 20, wherein the ischemia and / or reperfusion injury is cerebral, cardiac, pulmonary, or renal ischemia and / or reperfusion injury.

22. 21. The use according to claim 20, wherein the ischemia and / or reperfusion injury is cerebral ischemia, cerebral reperfusion injury or stroke.

23. 21. The use of claim 20, wherein (a) and (b) are administered parenterally.

24. 24. The use of claim 23, wherein (a) and (b) are administered intravenously.

25. 21. The use of claim 20, wherein (a) and (b) are administered during, before or after reperfusion.

26. 21. The use of claim 20, comprising administering (a) and (b) simultaneously to a subject.

27. (a) glibenclamide; (b) (i) exenatide, or a pharmaceutically acceptable salt thereof, or a structural or functional analog of exenatide selected from lixisenatide, albiglutide, liraglutide, taspoglutide, and dulaglutide (LY2189265); and (ii) canrenoate potassium.

10. A method for treating and / or preventing ischemia and / or reperfusion injury in an ex vivo organ before or during transplantation, comprising administering to a subject in need thereof a combination comprising:

28. The therapeutic and / or prophylactic agent according to claim 27, wherein the ischemia and / or reperfusion injury is cerebral ischemia, cerebral reperfusion injury, or stroke.

29. The therapeutic and / or prophylactic agent according to claim 27 or 28, wherein (a) and (b) are administered before transplantation.

30. 6. A combination according to claim 1 or 2, or a pharmaceutical composition according to claim 4 or 5, for use in the treatment and / or prevention of stroke.

31. 4. The pharmaceutical composition according to claim 3 for use in the treatment and / or prevention of stroke.

32. 6. A combination according to claim 1 or 2 or a pharmaceutical composition according to claim 4 or 5 for use in the treatment and / or prevention of a neurodegenerative disease.

33. 4. The pharmaceutical composition according to claim 3 for use in the treatment and / or prevention of neurodegenerative diseases.

34. 6. A combination according to claim 1 or 2, or a pharmaceutical composition according to any of claims 4 or 5, for use in the treatment and / or prevention of a neurodegenerative disease selected from Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis and vascular dementia.

35. 4. The pharmaceutical composition according to claim 3, for use in the treatment and / or prevention of a neurodegenerative disease selected from Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis and vascular dementia.

36. 6. A combination according to claim 1 or 2, or a pharmaceutical composition according to claim 4 or 5, for use in providing neuroprotection.

37. 4. The pharmaceutical composition of claim 3 for use in providing neuroprotection.

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