WO2025104674A1 - Combination therapy for treating neurodegenerative disorders - Google Patents

Abstract

The present disclosure provides a pharmaceutical composition for the prevention or treatment of neurodegenerative disorders comprising a phosphodiesterase 5 inhibitor and a glucagon-like peptide 1 (GLP-1) receptor agonist or a dipeptidyl peptidase IV (DPP-4) inhibitor as active ingredients. The phosphodiesterase 5 inhibitor is selected from mirodenafil, sildenafil, vardenafil, tadalafil, udenafil, dasantafil, avanafil, and pharmaceutically acceptable salts, solvates and hydrates thereof. The GLP-1 receptor agonist is selected from semaglutide, exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, tirzepatide, cotadutide, and taspoglutide. The DPP-4 inhibitor is selected from sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, teneligliptin, alogliptin, evogliptin, and anagliptin. The combination exhibits synergistic effects in reducing intracellular Aβ42, IL1β, and TNFα levels compared to the individual compounds alone.

Description

COMBINATION THERAPY FOR TREATING NEURODEGENERATIVE DISORDERS

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority from Korean Provisional Application Serial No. 10-2023-0156962 filed on November 14, 2023, which is incorporated herein by reference in its entirety.

FIELD

[0002] The present teachings relate to methods and compositions for the treatment of neurodegenerative disorders.

INTRODUCTION

[0003] Neurodegenerative disorders is a broad term used to describe a decline in cognitive function severe enough to interfere with daily life. It is not a specific disease or disorder, but rather a group of symptoms affecting memory, thinking, and social abilities. The most common neurodegenerative disorder is dementia, which is caused by Alzheimer's disease in 60-80% of cases. Other types include vascular dementia, Lewy body dementia, and frontotemporal dementia.

[0004] As the global population ages, the prevalence of dementia is increasing rapidly. This trend poses significant challenges for healthcare systems, families, and societies worldwide. The progressive nature of neurodegenerative disorders leads to increasing dependency and need for care as the condition advances. Currently, there is no cure for most types of neurodegenerative disorders, and available treatments primarily focus on managing symptoms and slowing disease progression.

[0005] Existing pharmacological approaches for treating neurodegenerative disorders typically target specific neurotransmitter systems or attempt to address underlying pathological processes. However, the complex and multifaceted nature of neurodegenerative disorders often limits the effectiveness of single-target therapies. This has led to growing interest in combination therapies that may address multiple aspects of the disease simultaneously.

[0006] Recent research has explored various molecular pathways and cellular mechanisms involved in the development and progression of neurodegenerative disorders. These investigations have highlighted potential new targets for therapeutic intervention. Among these, certain enzymes and receptors have emerged as promising candidates for drug development.

[0007] Despite advances in understanding the underlying mechanisms of neurodegenerative disorders, translating this knowledge into effective treatments remains challenging. Many potential therapies that show promise in preclinical studies fail to demonstrate significant benefits in clinical trials. This underscores the need for innovative approaches to drug discovery and development in the field of neurodegenerative disorder research.

[0008] The search for more effective treatments for neurodegenerative disorders continues to be an active area of scientific inquiry. Researchers are exploring various strategies, including repurposing existing drugs, developing novel compounds, and investigating combination therapies. The ultimate goal is to find interventions that can significantly improve cognitive function, slow disease progression, and enhance the quality of life for individuals affected by neurodegenerative disorders.

SUMMARY

[0009] According to an aspect of the present disclosure, a pharmaceutical composition is provided for the prevention or treatment of neurodegenerative disorders, comprising: a PDE-5 inhibitor, a GLP-1 receptor agonist, a GIP agonist, and glucagon wherein the molar ratio of any individual member of the group above to another member is between about (1,0,0,1000) to about (1000,0,0,0). Such molar ratios are described more fully below.

[0010] According to another aspect of the present disclosure, a pharmaceutical composition is provided for the prevention or treatment of neurodegenerative disorders, comprising: a PDE-5 inhibitor, a DPP-4 inhibitor, a GIP agonist, and glucagon wherein the molar ratio of any individual member of the group above to another member is between about (1,0,0,1000) to about (1000,0,0,0).

[0011] In various aspects, the PDE-5 inhibitor may be mirodenafil, sildenafil, vardenafil, tadalafil, udenafil, dasantafil, avanafil, and pharmaceutically acceptable salts, solvates and hydrates thereof, and in further aspects is mirodenafil. In various aspects, the GLP-1 receptor agonist may be semaglutide, exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, tirzepatide, cotadutide, and taspoglutide, and in further aspects is semaglutide. In various aspects, the DPP-4 inhibitor is sitagliptin. [0012] According to further aspects of the present disclosure, the neurodegenerative disorder may be Parkinson's disease (PD); sporadic or heritable dementia with Lewy bodies (DLB); pure autonomic failure (PAF) with aS deposition; multiple system atrophy (MSA); hereditary neurodegeneration with brain iron accumulation; and incidental Lewy body disease of advanced age; Alzheimer's disease of the Lewy body variant; Down's syndrome; progressive supranuclear palsy; essential tremor with Lewy bodies; familial parkinsonism with or without dementia resulting from a mutant gene and loci where no gene mutation has yet been identified; Creutzfeldt Jakob disease; bovine spongiform encephalopathy; secondary Parkinson disease/parkinsonism resulting from neurotoxin exposure/drug-induced parkinsonism with a-synuclein deposition; sporadic or heritable spinocerebellar ataxia; amyotrophic lateral sclerosis (ALS); idiopathic rapid eye movement sleep behavior disorder; and other conditions associated with central and/or peripheral a-synuclein accumulation in mammals accompanying a primary disease process.

[0013] According to further aspects of the present disclosure, the neurodegenerative disorder may be Alzheimer’s disease, Vascular dementia, Frontotemporal dementia (FTD), Lewy body dementia (LBD), Mixed dementia, Posterior cortical atrophy (PCA), Primary progressive aphasia (PPA), Corticobasal degeneration (CBD), and Progressive supranuclear palsy (PSP), idiopathic myeloma, amyloid polyneuropathy, amyloid cardiomyopathy, systemic senile amyloidosis, amyloid polyneuropathy, hereditary cerebral hemorrhage with amyloidosis, Down's syndrome, Scrapie, medullary carcinoma of the thyroid, isolated atrial amyloidosis, Pi-microglobulin amyloidosis, inclusion body myositis, muscle wasting disease, Islets of Langerhans diabetes, Type 1 diabetes, insulinoma, Type 2 diabetes mellitus, hereditary cerebral hemorrhage amyloidosis (Dutch), amyloid A (reactive) amyloidosis, secondary amyloidosis, familial Mediterranean fever, familial amyloid nephropathy with urticaria and deafness (Muckle-wells Syndrome), amyloid lambda L-chain amyloidosis, amyloid kappa L-chain amyloidosis, idiopathic associated amyloidosis, myeloma- associated amyloidosis, macroglobulinemia-associated amyloidosis, A beta 2M amyloidosis (chronic hemodialysis), ATTR amyloidosis (familial amyloid polyneuropathy (Portuguese, Japanese, Swedish)), familial amyloid cardiomyopathy (Danish), isolated cardiac amyloidosis, systemic senile amyloidosis, AIAPP or amylin insulinoma, atrial natriuretic factor amyloidosis (isolated atrial amyloidosis), procalcitonin amyloidosis (medullary carcinoma of the thyroid), gelsolin amyloidosis (familial amyloidosis (Finnish)), cystatin C (hereditary cerebral hemorrhage with amyloidosis (Icelandic)), AApo-A-I amyloidosis (familial amyloidotic polyneuropathy-Iowa), AApo-A-II amyloidosis, traumatic brain injury, fibrinogen- associated amyloidosis, Creutzfeldt- Jakob disease, Gertsmann-Straussler-Scheinker syndrome, bovine spongiform encephalitis, condition associated with homozygosity for the apolipoprotein E4 allele, and Huntington's disease.

[0014] In particular, the neurodegenerative disorder may be Parkinson's Disease (PD), Parkinson's Disease Dementia (PDD), Dementia with Lewy Bodies (DLB), Multiple System Atrophy (MSA), Pure Autonomic Failure (PAF), and Lewy Body Variant of Alzheimer's Disease (LBV). In particular, the neurodegenerative disorder may be Alzheimer’s disease, Vascular dementia, Frontotemporal dementia (FTD), Lewy body dementia (LBD), Mixed dementia, Posterior cortical atrophy (PCA), Primary progressive aphasia (PPA), Corticobasal degeneration (CBD), and Progressive supranuclear palsy (PSP). In various aspects, the neurodegenerative disorder may be Dementia with Lewy Bodies and Alzheimer’s disease.

[0015] In various aspects, the pharmaceutical composition is formulated for oral administration. In particular, the pharmaceutical composition is in the form of a tablet or capsule.

[0016] Various methods are provided for treating a neurodegenerative disorder in a subject in need thereof, the method comprising: administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a PDE-5 inhibitor, a GLP-1 receptor agonist, a GIP agonist, and glucagon, wherein the molar ratio of any individual member of the group above to another member is between about (1,0,0,1000) to about (1000,0,0,0).

[0017] In various further methods are provided for treating a neurodegenerative disorder in a subject in need thereof, the method comprising: administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a PDE-5 inhibitor, a DPP-4 inhibitor, a GIP agonist, and glucagon, wherein the molar ratio of any individual member of the group above to another member is between about (1,0,0,1000) to about (1000,0,0,0).

[0018] In various aspects, the PDE-5 inhibitor may be mirodenafil, sildenafil, vardenafil, tadalafil, udenafil, dasantafil, avanafil, and pharmaceutically acceptable salts, solvates and hydrates thereof, and in particular mirodenafil. In various aspects, the GLP-1 receptor agonist may be semaglutide, exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, tirzepatide, cotadutide, and taspoglutide, and in particular is semaglutide. In various aspects, the DPP-4 inhibitor may be sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, teneligliptin, alogliptin, evogliptin, and anagliptin, and in particular sitagliptin.

[0019] In further aspects, the neurodegenerative disorder is selected from the group consisting of Parkinson's disease (PD); sporadic or heritable dementia with Lewy bodies (DLB); pure autonomic failure (PAF) with aS deposition; multiple system atrophy (MSA); hereditary neurodegeneration with brain iron accumulation; and incidental Lewy body disease of advanced age; Alzheimer's disease of the Lewy body variant; Down's syndrome; progressive supranuclear palsy; essential tremor with Lewy bodies; familial parkinsonism with or without dementia resulting from a mutant gene and loci where no gene mutation has yet been identified; Creutzfeldt Jakob disease; bovine spongiform encephalopathy; secondary Parkinson disease/parkinsonism resulting from neurotoxin exposure/drug-induced parkinsonism with a-synuclein deposition; sporadic or heritable spinocerebellar ataxia; amyotrophic lateral sclerosis (ALS); idiopathic rapid eye movement sleep behavior disorder; and other conditions associated with central and/or peripheral a-synuclein accumulation in mammals accompanying a primary disease process.

[0020] In yet other aspects, the neurodegenerative disorder is selected from the group consisting of Alzheimer’s disease, Vascular dementia, Frontotemporal dementia (FTD), Lewy body dementia (LBD), Mixed dementia, Posterior cortical atrophy (PCA), Primary progressive aphasia (PPA), Corticobasal degeneration (CBD), and Progressive supranuclear palsy (PSP), idiopathic myeloma, amyloid polyneuropathy, amyloid cardiomyopathy, systemic senile amyloidosis, amyloid polyneuropathy, hereditary cerebral hemorrhage with amyloidosis, Down's syndrome, Scrapie, medullary carcinoma of the thyroid, isolated atrial amyloidosis, Pi-microglobulin amyloidosis, inclusion body myositis, muscle wasting disease, Islets of Langerhans diabetes, Type 1 diabetes, insulinoma, Type 2 diabetes mellitus, hereditary cerebral hemorrhage amyloidosis (Dutch), amyloid A (reactive) amyloidosis, secondary amyloidosis, familial Mediterranean fever, familial amyloid nephropathy with urticaria and deafness (Muckle-wells Syndrome), amyloid lambda L-chain amyloidosis, amyloid kappa L-chain amyloidosis, idiopathic associated amyloidosis, myeloma-associated amyloidosis, macroglobulinemia- associated amyloidosis, A beta 2M amyloidosis (chronic hemodialysis), ATTR amyloidosis (familial amyloid polyneuropathy (Portuguese, Japanese, Swedish)), familial amyloid cardiomyopathy (Danish), isolated cardiac amyloidosis, systemic senile amyloidosis, AIAPP or amylin insulinoma, atrial natriuretic factor amyloidosis (isolated atrial amyloidosis), procalcitonin amyloidosis (medullary carcinoma of the thyroid), gelsolin amyloidosis (familial amyloidosis (Finnish)), cystatin C (hereditary cerebral hemorrhage with amyloidosis (Icelandic)), AApo-A-I amyloidosis (familial amyloidotic polyneuropathy-Iowa), AApo-A-II amyloidosis, traumatic brain injury, fibrinogen- associated amyloidosis, Creutzfeldt- Jakob disease, Gertsmann-Straussler-Scheinker syndrome, bovine spongiform encephalitis, condition associated with homozygosity for the apolipoprotein E4 allele, and Huntington's disease.

[0021 ] In various aspects, the neurodegenerative disorder may be Parkinson's Disease (PD), Parkinson's Disease Dementia (PDD), Dementia with Lewy Bodies (DLB), Multiple System Atrophy (MSA), Pure Autonomic Failure (PAF), and Lewy Body Variant of Alzheimer's Disease (LBV). In various aspects, the neurodegenerative disorder may be Alzheimer’s disease, Vascular dementia, Frontotemporal dementia (FTD), Lewy body dementia (LBD), Mixed dementia, Posterior cortical atrophy (PCA), Primary progressive aphasia (PPA), Corticobasal degeneration (CBD), and Progressive supranuclear palsy (PSP). In further aspects, the neurodegenerative disorder may be Dementia with Lewy Bodies and Alzheimer’s disease.

[0022] These and other features, aspects and advantages of the present teachings will become better understood with reference to the following description, examples and appended claims.

DRAWINGS

[0023] Those of skill in the art will understand that the drawings, described below, are for illustrative purposes only. The drawings are not intended to limit the scope of the present teachings in any way.

[0024] FIG. 1 illustrates a bar graph showing effects of AR1001 and semaglutide on IL-6 levels, according to aspects of the present disclosure.

[0025] FIG. 2 illustrates a bar graph showing effects of AR1001 and semaglutide on IL-6 levels, according to aspects of the present disclosure.

[0026] FIG. 3 illustrates a bar graph showing effects of AR1001 and Tirzepatide on IL-6 levels, according to aspects of the present disclosure.

[0027] FIG. 4 illustrates a bar graph showing effects of AR1001 and dulaglutide on IL-6 levels, according to aspects of the present disclosure.

[0028] FIG. 5 illustrates a bar graph showing effects of AR1001 and semaglutide on IL-6 levels, according to aspects of the present disclosure.

[0029] FIG. 6 illustrates a bar graph showing effects of AR1001 and Tirzepatide on IL-6 levels, according to aspects of the present disclosure. [0030] FIG. 7 illustrates a bar graph showing effects of AR1001 and dulaglutide on IL-6 levels, according to aspects of the present disclosure.

[0031] FIG. 8 illustrates bar graphs showing effects of AR1001 and Exenatide on IL-6 levels, according to aspects of the present disclosure.

[0032] FIG. 9 illustrates a bar graph showing effects of AR1001 and semaglutide on JC-1 ratio, according to an embodiment.

[0033] FIG. 10 illustrates a bar graph showing effects of AR1001 and Exenatide on JC-1 ratio, according to an embodiment.

[0034] FIG. 11 illustrates a bar graph showing effects of AR1001 and Semaglutide on Ap reduction, according to an embodiment.

[0035] FIG. 12 illustrates a bar graph showing effects of AR1001 and Semaglutide on ILip reduction, according to an embodiment.

[0036] FIG. 13 illustrates a bar graph showing effects of AR1001 and Semaglutide on TNFa reduction, according to an embodiment.

[0037] FIG. 14 illustrates a bar graph and data table showing effects of AR1001 and Sitagliptin on Ap reduction, according to an embodiment.

[0038] FIG. 15 illustrates a bar graph and data table showing effects of AR1001 and Vildagliptin on Ap reduction, according to aspects of the present disclosure.

[0039] FIG. 16 illustrates a bar graph and data table showing effects of AR1001 and Linagliptin on Ap reduction, according to an embodiment.

[0040] FIG. 17 illustrates a bar graph showing effects of AR1001 and Sitagliptin on ILip levels, according to aspects of the present disclosure.

[0041] FIG. 18 illustrates a bar graph showing effects of AR1001 and Sitagliptin on ILip reduction, according to an embodiment.

[0042] FIG. 19 illustrates a bar graph showing effects of AR1001 and Sitagliptin on TNFa levels, according to aspects of the present disclosure.

[0043] FIG. 20 illustrates a bar graph showing effects of AR1001 and Sitagliptin on TNFa reduction, according to an embodiment.

[0044] FIG. 21 illustrates a bar graph showing effects of AR1001 and Linagliptin on ILip reduction, according to aspects of the present disclosure.

[0045] FIG. 22 illustrates a bar graph showing effects of AR1001 and Linagliptin on TNFa reduction, according to an embodiment.

DETAILED DESCRIPTION [0046] Abbreviations and Definitions

[0047] To facilitate understanding of the invention, a number of terms and abbreviations as used herein are defined below as follows:

[0048] All patents, applications, published applications and other publications cited herein are incorporated by reference in their entirety. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the invention belongs. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts. Should a discrepancy exist between a depicted structure and a name given for that structure, the depicted structure is to be accorded more weight. Where the stereochemistry of a structure or a portion of a structure is not indicated in a depicted structure or a portion of the depicted structure, the depicted structure is to be interpreted as encompassing all of its possible stereoisomers.

[0049] Any methods, devices and materials similar or equivalent to those described herein can be used in the practice of this invention. The following definitions are provided to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure. In the event that there is a plurality of definitions for a term herein, those in this section prevail unless stated otherwise. Headings used herein are for organizational purposes only and in no way limit the invention described herein.

[0050] The term “administering” refers to the act of delivering a combination or composition described herein into a subject by such routes as oral, mucosal, topical, suppository, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal or subcutaneous administration. Parenteral administration includes intravenous, intramuscular, intra-arteriole, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial administration. Administration generally occurs after the onset of the disease, disorder, or condition, or its symptoms but, in certain instances, can occur before the onset of the disease, disorder, or condition, or its symptoms (e.g., administration for patients prone to such a disease, disorder, or condition).

[0051] The terms “coadministering” or “coadministration” or “combination” refers to administration of two or more agents (e.g., a combination described herein including, optionally, another active agent such as an anti-neurodegenerative agent described herein). The timing of coadministration depends in part of the combination and individual compositions administered and can include administration at the same time, just prior to, or just after the administration of one or more additional therapies, for example therapies such as an anti-neurodegenerative agent including, for example, immunotherapy. Coadministration is meant to include simultaneous or sequential administration of each compound of the combination. Thus, the preparations can also be combined, when desired, with other active substances (e.g., to reduce metabolic degradation). The compounds described herein can be used in combination with one another, with other active agents known to be useful in treating neurodegeneration.

[0052] The terms “therapies” and “therapy” refer to any protocol(s), method(s), and/or agent(s) that can be used in the prevention, treatment, management, and/or amelioration of a disease, disorder, or condition or one or more symptoms thereof. In certain instances the term refers to active agents such as an anti-neurodegenerative agent described herein. The terms “therapy” and “therapy” can also refer to anti-viral therapy, anti-bacterial therapy, anti-fungal therapy, anti-neurodegenerative therapy, biological therapy, supportive therapy, and/or other therapies useful in treatment, management, prevention, or amelioration of a disease, disorder, or condition or one or more symptoms thereof known to one skilled in the art, for example, a medical professional such as a physician.

[0053] The term “patient” or “subject” refers to a mammal, such as a human, bovine, rat, mouse, dog, monkey, ape, goat, sheep, cow, or deer. Generally a patient as described herein is human. The subject according to the present disclosure may be a human subject suffering from Alzheimer's disease (e.g., AD, Alzheimer's type dementia) or a human subject possibly suffering from Alzheimer's disease. In these cases, whether a human subject is suffering from or likely to suffer from Alzheimer's disease may be determined by the methods disclosed in the Examples provided herein, and by methods commonly practiced by those skilled in the art. Human subjects suffering from or potentially suffering from Alzheimer's disease include, for example, those for whom a cognitive function test such as the Clinical Dementia Rating Scale Sum of Boxes (CDR-SB), the Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-Cog), or the Mini-Mental State Examination (MMSE) suggests a decline in cognitive function or the possibility thereof. The subject may be a human subject with brain deposits or cerebrospinal fluid concentrations of proteins known to accumulate in Alzheimer's disease patients, such as amyloid-P and tau proteins, a human subject with decreased metabolic function of the brain or atrophy of brain tissue.

[0054] The terms “treating” or “treatment” refer to any indicia of success or amelioration of the progression, severity, and/or duration of a disease, pathology or condition, including any objective or subjective parameter such as abatement; remission; diminishing of symptoms or making the injury, pathology or condition more tolerable to the patient; slowing in the rate of degeneration or decline; making the final point of degeneration less debilitating; or improving a patient’s physical or mental well-being.

[0055] The terms “AR1001” and “mirodenafil” refer to the chemical compound, 5- ethyl-2- [5- [4-(2-hydroxyethyl)piperazin- 1 -yl] sulfonyl-2-propoxyphenyl] -7 -propyl -3 7- pyrrolo[3,2-d]pyrimidin-4-one, which has the following chemical formula:

Mirodenafil is a member of the class of pyrrolopyrimidines that is 3,5-dihydro-4H- pyrrolo[3,2-d]pyrimidin-4-one having a 5- { [4-(2-hydroxyethyl)piperazin-l-yl] sulfonyl} -2- propoxyphenyl group at positon 2, ethyl group at position 5, and a propyl group at position 7. It is a phosphodiesterase type 5 inhibitor which is used for the treatment of erectile dysfunction. It has a role as an EC 3.1.4.35 (3',5'-cyclic-GMP phosphodiesterase) inhibitor and a vasodilator agent. It is a sulfonamide, a pyrrolopyrimidine, a N-alkylpiperazine, a primary alcohol and an aromatic ether. Mirodenafil has been used in trials studying the treatment and supportive care of Kidney Diseases, Urologic Diseases, Renal Insufficiency, Erectile Dysfunction, and Male Erectile Dysfunction. More information about mirodenafil can be found in U.S. Pat. No. 9,750,743, and at Kang, B.W., Kim, F., Cho, JY. et al. Phosphodiesterase 5 inhibitor mirodenafil ameliorates Alzheimer-like pathology and symptoms by multimodal actions. Alz Res Therapy 14, 92 (2022). https://doi.org/10.1186/sl3195-022-01034-3. Mirodenafil is commercially available from vendors including Cooke Chemical Co., Ltd. (#M4050735), AstaTech, Inc. (#C16519), and RR Scientific (#R772058), among others.

[0056] The term “anti-neurodegenerative agent” is used in accordance with its plain ordinary meaning and refers to a composition having neuronal protective properties or the ability to inhibit degeneration of neuronal cells. The term can include neuroprotective agents known to those of skill in the art. In embodiments, an anti-neurodegenerative agent is an agent identified herein having utility in methods of treating neurodegeneration. In embodiments, an anti-neurodegenerative agent is an agent approved by the US FDA or similar regulatory agency of a country other than the US, for treating neurodegeneration. This term includes, but is not limited to, the combinations described herein, and in the context of the present disclosure other anti-neurodegenerative agents.

[0057] The term “neurodegenerative disorder” or “neurodegeneration” refers to a condition primarily characterized by neuron (z.e., neuronal cell) loss. The most common neurodegenerative disorders include Alzheimer’s disease and Parkinson’s disease. Although there are several medicines currently approved for managing neurodegenerative disorders, a large majority of them only help with associated symptoms. A listing of over 500 Neurological Disorders is provided by the US National Institute of Neurological Disorders and Stroke (NINDS) currently available at: https://www.ninds.nih.gov/health- information/disorders. See also, Wolfe, M. The Molecular and Cellular Basis of Neurodegenerative Diseases: Underlying Mechanisms. Academic Press, April 18, 2018; Lewis, P. The Molecular and Clinical Pathology of Neurodegenerative Disease. Academic Press, December 14, 2018. For the purposes of the present disclosure, the term “neurodegenerative disorders” includes those disorders, and specifically the synucleinopathies and amyloidoses and amyloidosis-associated condition described herein.

[0058] The term “alpha-synuclein” refers to a protein or polypeptide (aS or aS protein), as used herein, includes a single, monomeric protein or polypeptide, as well as such aS proteins and polypeptides in the form of oligomers, e.g., in the form of dimers, trimers, tetramers, or in the form of lipid- associated complexes, or lipid-free forms, or in the form of aggregates, and any of these forms can be soluble or insoluble. The terms also include the aS proteins found in complexes with other molecules.

[0059] The term “synucleinopathy” is used herein to name a group of neurodegenerative disorders characterized by the presence of increased levels, e.g., steadystate levels, of any one or more of soluble non-fibrillary variants, soluble oligomeric isoforms, insoluble non-fibrillary variants, complexes, and insoluble fibrillary aggregates of aS protein within cellular compartments of selective populations of neurons and glia. Thus, the aS steady-state level is understood to encompass all soluble as well as insoluble and intermediate (metastable) forms of the SNCA gene product. See, UniProt P37840; Gene ID: 6622; NCBI Reference Sequence: NG_011851.1. These disorders include any one of the following grouped as “invariable” (or “primary”) synucleinopathies: Parkinson's disease (PD) e.g., sporadic Parkinson disease/parkinsonism and familial Parkinson disease/parkinsonism; sporadic or heritable dementia with Lewy bodies (DLB) (aka diffuse Lewy body disease); pure autonomic failure (PAF) with aS deposition; multiple system atrophy (MSA) (of cerebellar, parkinsonian, or mixed type); hereditary neurodegeneration with brain iron accumulation (aka, Hallervordem Spatz disease or pantothenate kinase 2-linked neurodegeneration); and incidental Lewy body disease of advanced age. Furthermore, “variable” (or “secondary”) synucleinopathies have been identified, where dysregulation of the alpha-synuclein metabolism is recognized to be a secondary event (given the abundance of the protein in the nervous system), which nevertheless contributes significantly to the course, penetrance, age-of-onset, severity and expressivity of the primary illness. Disorders with variable synucleinopathy include, but are not limited to, Alzheimer's disease of the Lewy body variant; Down's syndrome; progressive supranuclear palsy; essential tremor with Lewy bodies; familial parkinsonism with or without dementia resulting from a mutant gene and loci where no gene mutation has yet been identified; Creutzfeldt Jakob disease and related prion diseases such as bovine spongiform encephalopathy (mad cow disease); secondary Parkinson disease/parkinsonism resulting from neurotoxin exposure/drug-induced parkinsonism with a-synuclein deposition; sporadic or heritable spinocerebellar ataxia; amyotrophic lateral sclerosis (ALS); idiopathic rapid eye movement sleep behavior disorder; and other conditions associated with central and/or peripheral a-synuclein accumulation in mammals accompanying a primary disease process. See, Schlossmacher MG a-synuclein and synucleinopathies. The Dementias 2 Blue Books of Practical Neurology; Editors: Growdon J H & Rossor M N. Butterworth Heinemann, Inc., Oxford. 2007; Chapter 8: pp 184-213. Clinically, all of these related disorders are characterized by a chronic and progressive decline in motor, cognitive, behavioral, and/or autonomic functions, depending on the distribution of the alpha-synuclein abnormalities.

[0060] The terms “amyloidosis” and “amyloidoses” refer to a group of diseases that involve the accumulation of amyloid proteins in the body. Amyloid proteins can be deposited in one part of the body, called localized amyloidosis, or in multiple parts, called systemic amyloidosis. Many forms of amyloidosis exist, and the disease can be classified into four groups: primary amyloidosis, secondary amyloidosis, hereditary amyloidosis, and amyloidosis associated with normal aging. Primary amyloidosis (light chain amyloidosis) occurs with abnormalities of plasma cells, and some people with primary amyloidosis also have multiple myeloma (cancer of the plasma cells). Typical sites of amyloid buildup in primary amyloidosis are the heart, lungs, skin, tongue, thyroid gland, intestines, liver, kidneys, and blood vessels. Secondary amyloidosis may develop in response to various diseases that cause persistent infection or inflammation, such as tuberculosis, rheumatoid arthritis, and familial Mediterranean fever. Typical sites of amyloid buildup in secondary amyloidosis are the spleen, liver, kidneys, adrenal glands, and lymph nodes. Hereditary amyloidosis has been noted in some families, particularly those from Portugal, Sweden, and Japan. The amyloid-producing defect occurs because of mutations in specific proteins in the blood. Typical sites for amyloid buildup in hereditary amyloidosis are the nerves, heart, blood vessels, and kidneys. Alzheimer's disease is a type of localized amyloidosis where amyloidbeta proteins build up in the brain. This is the most common type of amyloidosis in humans and the most common form of dementia. The “amyloid hypothesis” is the prevailing theory that Alzheimer's disease is caused by the accumulation of beta-amyloid proteins in the brain. Some studies have shown that amyloid triggers a binding of two proteins in the brain's neurons, which can lead to the rapid accumulation of tau proteins. Tau proteins are a primary driver of neurodegeneration in Alzheimer's disease. Amyloidosis can be caused by chronic inflammation or genetic mutation. There are many different types of amyloid proteins involved in amyloidosis, and each type of amyloid deposit can characterize a different disease.

[0061] The term “amyloidosis-associated condition” refers to a disease that is associated with amyloid deposition and can include but not be limited to Alzheimer's Disease, mild cognitive impairment due to Alzheimer's disease, mild Alzheimer's disease dementia, prodromal stage of Alzheimer’s disease, early-onset Alzheimer’s disease, mild Alzheimer’s disease, moderate Alzheimer’s disease, early Alzheimer’s disease, preclinical Alzheimer’s disease, idiopathetic myeloma, amyloid polyneuropathy, amyloid cardiomyopathy, systemic senile amyloidosis, amyloid polyneuropathy, hereditary cerebral hemorrhage with amyloidosis, Down's syndrome, Scrapie, medullary carcinoma of the thyroid, isolated atrial amyloid, p2-microglobulin amyloid in dialysis patients, inclusion body myositis, P2-amyloid deposits in muscle wasting disease, and Islets of Langerhans diabetes Type II insulinoma, Type 2 diabetes mellitus, hereditary cerebral hemorrhage amyloidosis (Dutch), amyloid A (reactive), secondary amyloidosis, familial Mediterranean fever, familial amyloid nephropathy with urticaria and deafness (Muckle-wells Syndrome), amyloid lambda L-chain or amyloid kappa L-chain (idiopathic, myeloma or macroglobulinemia-associated) A beta 2M (chronic hemodialysis), ATTR (familial amyloid polyneuropathy (Portuguese, Japanese, Swedish)), familial amyloid cardiomyopathy (Danish), isolated cardiac amyloid, systemic senile amyloidoses, AIAPP or amylin insulinoma, atrial naturetic factor (isolated atrial amyloid), procalcitonin (medullary carcinoma of the thyroid), gelsolin (familial amyloidosis (Finnish)), cystatin C (hereditary cerebral hemorrhage with amyloidosis (Icelandic)), AApo- A-l (familial amyloidotic polyneuropathy-Iowa), AApo-A-II (accelerated senescence in mice), head injuries (traumatic brain injury), dementia, fibrinogen-associated amyloid; and Asor or Pr P-27 (scrapie, Creutzfeld Jacob disease, Gertsmann-Straussler-Scheinker syndrome, bovine spongiform encephalitis) or in cases of persons who are homozygous for the apolipoprotein E4 allele, and the condition associated with homozygosity for the apolipoprotein E4 allele or Huntington's disease.

[0062] The term “early-onset Alzheimer’s disease” or “EOAD” refers to AD cases where symptoms appear before age 65. EOAD includes, but is not limited to: Genetic (Familial) Alzheimer's Disease and its phenotypic variants including Logopenic Variant Primary Progressive Aphasia, Posterior Cortical Atrophy, Behavioral/Dysexecutive Alzheimer's Disease, and Acalculia Variant.

[0063] As used herein, the term “pharmaceutically acceptable” means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0064] As used herein, the term “pharmaceutically acceptable carrier” refers to a diluent, adjuvant, excipient, or vehicle with which a compound is administered. Such carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like, polyethylene glycols, glycerine, propylene glycol, or other synthetic solvents. Water is a preferred carrier when a compound is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, and the like. A compound, if desired, can also combine minor amount of wetting or emulsifying agents, or pH buffering agents such as acetates, citrates, or phosphates. Antibacterial agents such as a benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid; and agents for the adjustment of tonicity such as sodium chloride or dextrose may also be a carrier. Methods for producing compounds in combination with carriers are known to those of skill in the art.

[0065] As used herein, the term “pharmaceutically acceptable salt” includes those salts of a pharmaceutically acceptable compound formed with free amino groups such as those derived from hydrochloric, phosphoric, acetic, oxalic, and tartaric acids, and those formed with free carboxyl groups such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamine, triethylamine, 2-ethylamino ethanol, histidine, and procaine. If the compound is basic, salts may be prepared from pharmaceutically acceptable non-toxic acids including inorganic and organic acids. Such acids include acetic, benzene-sulfonic (besylate), benzoic, camphorsulfonic, citric, ethenesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic, and the like. Particularly preferred are besylate, hydrobromic, hydrochloric, phosphoric, and sulfuric acids. If the compound is acidic, salts may be prepared from pharmaceutically acceptable organic and inorganic bases. Suitable organic bases include, but are not limited to, lysine, N,N’ -dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylene diamine, meglumine (N-methyl-glucamine) and procaine. Suitable inorganic bases include, but are not limited to, alkaline and earth- alkaline metals such as aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc. Methods for synthesizing such salts are known to those of skill in the art.

[0066] The term “effective amount” refers to the amount of a therapy (e.g., a combination provided herein or another active agent such as an anti-neurodegenerative agent described herein) which is sufficient to accomplish a stated purpose or otherwise achieve the effect for which it is administered. An effective amount can be sufficient to reduce and/or ameliorate the progression, development, recurrence, severity and/or duration of a given disease, disorder or condition and/or a symptom related thereto. An effective amount can be a “therapeutically effective amount” which refers to an amount sufficient to provide a therapeutic benefit such as, for example, the reduction or amelioration of the advancement or progression of a given disease, disorder or condition, reduction or amelioration of the recurrence, development or onset of a given disease, disorder or condition, and/or to improve or enhance the prophylactic or therapeutic effect(s) of another therapy. A therapeutically effective amount of a composition described herein can enhance the therapeutic efficacy of another therapeutic agent.

[0067] The term “therapeutically effective amount” refers to the amount of a therapy which is sufficient to accomplish a stated purpose or otherwise achieve the effect for which it is administered. An effective amount can be sufficient to reduce and/or ameliorate the progression, development, recurrence, severity and/or duration of a given disease, disorder or condition and/or a symptom related thereto. An effective amount can be a “therapeutically effective amount” which refers to an amount sufficient to provide a therapeutic benefit such as, for example, the reduction or amelioration of the advancement or progression of a given disease, disorder or condition, reduction or amelioration of the recurrence, development or onset of a given disease, disorder or condition, and/or to improve or enhance the prophylactic or therapeutic effect(s) of another therapy. A therapeutically effective amount of a composition described herein can enhance the therapeutic efficacy of another therapeutic agent.

[0068] The term “regimen” refers to a protocol for dosing and timing the administration of one or more therapies (e.g., combinations described herein including, optionally, another active agent such as an anti-neurodegenerative agent described herein) for treating a disease, disorder, or condition described herein. A regimen can include periods of active administration and periods of rest as known in the art. Active administration periods include administration of combinations and compositions described herein and the duration of time of efficacy of such combinations and compositions. Rest periods of regimens described herein include a period of time in which no compound is actively administered, and in certain instances, includes time periods where the efficacy of such compounds can be minimal. Combination of active administration and rest in regimens described herein can increase the efficacy and/or duration of administration of the combinations and compositions described herein.

[0069] The term “enhance” refers to an increase or improvement in the function or activity of a protein or cell after administration or contacting with a combination described herein compared to the protein or cell prior to such administration or contact.

[0070] Combination Therapies for the Treatment of Neurodegenerative Disorders

[0071] The present disclosure relates to pharmaceutical compositions and methods for the prevention or treatment of neurodegenerative disorders. More specifically, the present disclosure provides combination therapies utilizing (1) a phosphodiesterase 5 inhibitor (PDE- 5 inhibitor) in conjunction with (2) either a glucagon-like peptide 1 (GLP-1) receptor agonist or a dipeptidyl peptidase IV (DPP-4) inhibitor, (3) a Gastric inhibitory peptide (GIP; also known as “glucose-dependent insulinotropic polypeptide”) agonist, and (4) glucagon in molar ratios more fully described below.

[0072] Neurodegenerative disorders encompass a range of cognitive disorders characterized by impairment of memory, thinking, and social abilities. As the global population ages, the prevalence of neurodegenerative disorders continues to increase, posing significant challenges for healthcare systems and societies worldwide. Current treatments for neurodegenerative disorders primarily focus on managing symptoms and slowing disease progression, with limited success in addressing the underlying causes of cognitive decline. [0073] The pharmaceutical compositions described herein may provide a novel approach to treating neurodegenerative disorders by combining agents that target multiple pathways involved in the disease process. Phosphodiesterase 5 inhibitors may modulate signaling pathways related to neuronal function and survival. GLP-1 receptor agonists and DPP-4 inhibitors may influence metabolic processes and neuroprotective mechanisms in the brain.

[0074] In some aspects, the combination of a phosphodiesterase 5 inhibitor with a GLP- 1 receptor agonist may result in synergistic effects on reducing intracellular amyloid beta levels and inflammatory markers associated with dementia. The combination may also demonstrate improved efficacy compared to either agent alone in preserving mitochondrial function in neuronal cells.

[0075] In other aspects, the combination of a phosphodiesterase 5 inhibitor with a DPP-4 inhibitor may exhibit enhanced neuroprotective properties and anti-inflammatory effects. This combination may provide a multi-targeted approach to addressing the complex pathophysiology of neurodegenerative disorders.

[0076] The pharmaceutical compositions described herein may be formulated for various routes of administration and may contain specific ratios of active ingredients to optimize therapeutic effects. The combinations may be used to treat or prevent various forms of neurodegenerative disorders, including but not limited to Alzheimer's disease, vascular dementia, and Lewy body dementia.

[0077] By targeting multiple aspects of neurodegenerative disorders pathology simultaneously, these combination therapies may offer potential advantages in efficacy and disease modification compared to existing single-agent treatments. The disclosed compositions and methods may provide new options for addressing the growing global burden of neurodegenerative disorders and improving outcomes for affected individuals.

[0078] Gastric inhibitory peptide (GIP) and glucagon-like peptide- 1 (GLP-1) are two important incretin hormones that play crucial roles in glucose homeostasis and metabolism. GIP (Gastric Inhibitory Peptide) is a 42-amino acid peptide hormone secreted by K cells in the small intestine in response to nutrient ingestion. Its main functions include stimulating insulin secretion from pancreatic P-cells in a glucose-dependent manner, promoting P-cell proliferation and inhibiting apoptosis, enhancing postprandial glucagon response, facilitating fat deposition in adipose tissues, and promoting bone formation.

[0079] GLP-1 (Glucagon-Like Peptide- 1) is a 30- or 31 -amino acid peptide hormone produced by L-cells in the distal ileum and colon. Its primary functions are stimulating insulin secretion from pancreatic P-cells (glucose-dependent), inhibiting glucagon secretion from pancreatic a-cells, slowing gastric emptying and reducing appetite, promoting P-cell proliferation and inhibiting apoptosis, and inhibiting bone resorption,

[0080] Glucagon is a hormone produced by pancreatic a-cells that plays a crucial role in maintaining blood glucose levels. Its main functions include stimulating glycogenolysis (conversion of stored glycogen to glucose), promoting gluconeogenesis (production of glucose from non-carbohydrate sources), reducing glucose consumption by the liver, and stimulating breakdown of fat stores in adipose tissue.

[0081 ] Interplay Between GIP, GLP-1, and Glucagon

[0082] GIP and GLP- 1 work together to enhance insulin secretion and improve glucose control, while glucagon acts as a counterregulatory hormone to prevent hypoglycemia. Both GIP and GLP-1 promote P-cell proliferation and survival, expanding pancreatic P-cell mass. However, GIP enhances postprandial glucagon response, while GLP- 1 suppresses it. GLP-1 has a more pronounced effect on reducing appetite and food intake compared to GIP. Due to their complementary effects, dual GIP/GLP-1 receptor agonists are being developed as potential treatments for type 2 diabetes and obesity. These combination therapies have shown promising results in clinical trials, offering greater weight loss and better glycemic control than GLP- 1 receptor agonists alone.

[0083] In conclusion, GIP, GLP-1, and glucagon form a complex regulatory system for glucose homeostasis and metabolism. Understanding their interplay and developing therapies that target multiple hormones simultaneously may lead to more effective treatments for a variety of disorders. Quite unexpectedly, these disorders include neurodegenerative disorders which is a feature of the present invention. What is surprising is that the combination of a PDE-5 inhibitor or DPP-4 inhibitor within the complex of GIP, GLP-1, and glucagon modulators enhances the therapeutic effect of any one of the individual modulators in the regulatory system. Without being bound by a particular theory,

[0084] PDE-5 inhibitors

[0085] The phosphodiesterase 5 inhibitor may be selected from the group consisting of mirodenafil, sildenafil, vardenafil, tadalafil, udenafil, dasantafil, avanafil, and pharmaceutically acceptable salts, solvates and hydrates thereof.

[0086] DPP-4 inhibitors [0087] The DPP-4 inhibitor may be selected from the group consisting of Sitagliptin, Vildagliptin, Saxagliptin, Linagliptin, Gemigliptin, Teneligliptin, Alogiptin, Evogliptin, and Anagliptin.

[0088] GLP-1 receptor agonists

The GLP- 1 receptor agonist may be selected from the group consisting of semaglutide, exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, tirzepatide, cotadutide, and taspoglutide.

[0089] GIP agonists

[0090] The GIP agonist may be tirzepatide (which has dual activity as a GLP-1 receptor agonist), CT-288 (Roche), or other Research Use Only compounds known by those of skill in the art.

[0091 ] Glucagon

[0092] As described above, glucagon is a peptide hormone produced by the alpha cells in the pancreas that plays a crucial role in regulating blood glucose levels.

[0093] Dual Agonists and Triple Agonists

[0094] Those of skill in the art will recognize that some compounds may act on multiple pathways, such as tirzepatide with is a dual agonist related to GLP-1 receptor and GIP. In another example, Retatrutide and SAR441255 are triple agonists related to GLP-1 receptor, GIP, and glucagon.

[0095] Molar Ratios

[0096] In some aspects, the pharmaceutical composition may include a PDE-5 inhibitor, a GLP- 1 receptor agonist or DPP-4 inhibitor, a GIP agonist, and glucagon in specific molar ratios. The molar ratio of (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, and (4) glucagon in the pharmaceutical composition can include (100, 0, 0, 900), (100, 0, 100, 800), (100, 0, 200, 700), (100, 0, 300, 600), (100, 0, 400, 500), (100, 0, 500, 400), (100, 0, 600, 300), (100, 0, 700, 200), (100, 0, 800, 100), (100, 0, 900, 0), (100, 100, 0, 800), (100, 100, 100, 700), (100, 100, 200, 600), (100, 100, 300, 500), (100, 100, 400, 400), (100, 100, 500, 300), (100, 100, 600, 200), (100, 100, 700, 100), (100, 100, 800, 0), (100, 200, 0, 700), (100, 200, 100, 600), (100, 200, 200, 500), (100, 200, 300, 400), (100, 200, 400, 300), (100, 200, 500, 200), (100, 200, 600, 100), (100, 200, 700, 0), (100, 300, 0, 600), (100, 300, 100, 500), (100, 300, 200, 400), (100, 300, 300, 300), (100, 300, 400, 200), (100, 300, 500, 100), (100, 300, 600, 0), (100, 400, 0, 500), (100, 400, 100, 400), (100, 400, 200, 300), (100, 400, 300, 200), (100, 400, 400, 100), (100, 400, 500, 0), (100, 500, 0, 400), (100, 500, 100, 300), (100, 500, 200, 200), (100, 500, 300, 100), (100, 500, 400, 0), (100, 600, 0, 300), (100, 600, 100, 200), (100, 600, 200, 100), (100, 600, 300, 0), (100, 700, 0, 200), (100, 700, 100, 100), (100, 700, 200, 0), (100, 800, 0, 100), (100, 800, 100, 0), (100, 900, 0, 0), (200, 0, 0, 800), (200, 0, 100, 700), (200, 0, 200, 600), (200, 0, 300, 500), (200, 0, 400, 400), (200, 0, 500, 300), (200, 0, 600, 200), (200, 0, 700, 100), (200, 0, 800, 0), (200, 100, 0, 700), (200, 100, 100, 600), (200, 100, 200, 500), (200, 100, 300, 400), (200, 100, 400, 300), (200, 100, 500, 200), (200, 100, 600, 100), (200, 100, 700, 0), (200, 200, 0, 600), (200, 200, 100, 500), (200, 200, 200, 400), (200, 200, 300, 300), (200, 200, 400, 200), (200, 200, 500, 100), (200, 200, 600, 0), (200, 300, 0, 500), (200, 300, 100, 400), (200, 300, 200, 300), (200, 300, 300, 200), (200, 300, 400, 100), (200, 300, 500, 0), (200, 400, 0, 400), (200, 400, 100, 300), (200, 400, 200, 200), (200, 400, 300, 100), (200, 400, 400, 0), (200, 500, 0, 300), (200, 500, 100, 200), (200, 500, 200, 100), (200, 500, 300, 0), (200, 600, 0, 200), (200, 600, 100, 100), (200, 600, 200, 0), (200, 700, 0, 100), (200, 700, 100, 0), (200, 800, 0, 0), (300, 0, 0, 700), (300, 0, 100, 600), (300, 0, 200, 500), (300, 0, 300, 400), (300, 0, 400, 300), (300, 0, 500, 200), (300, 0, 600, 100), (300, 0, 700, 0), (300, 100, 0, 600), (300, 100, 100, 500), (300, 100, 200, 400), (300, 100, 300, 300), (300, 100, 400, 200), (300, 100, 500, 100), (300, 100, 600, 0), (300, 200, 0, 500), (300, 200, 100, 400), (300, 200, 200, 300), (300, 200, 300, 200), (300, 200, 400, 100), (300, 200, 500, 0), (300, 300, 0, 400), (300, 300, 100, 300), (300, 300, 200, 200), (300, 300, 300, 100), (300, 300, 400, 0), (300, 400, 0, 300), (300, 400, 100, 200), (300, 400, 200, 100), (300, 400, 300, 0), (300, 500, 0, 200), (300, 500, 100, 100), (300, 500, 200, 0), (300, 600, 0, 100), (300, 600, 100, 0), (300, 700, 0, 0), (400, 0, 0, 600), (400, 0, 100,

500), (400, 0, 200, 400), (400, 0, 300, 300), (400, 0, 400, 200), (400, 0, 500, 100), (400, 0,

600, 0), (400, 100, 0, 500), (400, 100, 100, 400), (400, 100, 200, 300), (400, 100, 300, 200), (400, 100, 400, 100), (400, 100, 500, 0), (400, 200, 0, 400), (400, 200, 100, 300), (400, 200, 200, 200), (400, 200, 300, 100), (400, 200, 400, 0), (400, 300, 0, 300), (400, 300, 100, 200), (400, 300, 200, 100), (400, 300, 300, 0), (400, 400, 0, 200), (400, 400, 100, 100), (400, 400, 200, 0), (400, 500, 0, 100), (400, 500, 100, 0), (400, 600, 0, 0), (500, 0, 0, 500), (500, 0, 100,

400), (500, 0, 200, 300), (500, 0, 300, 200), (500, 0, 400, 100), (500, 0, 500, 0), (500, 100, 0,

400), (500, 100, 100, 300), (500, 100, 200, 200), (500, 100, 300, 100), (500, 100, 400, 0), (500, 200, 0, 300), (500, 200, 100, 200), (500, 200, 200, 100), (500, 200, 300, 0), (500, 300, 0, 200), (500, 300, 100, 100), (500, 300, 200, 0), (500, 400, 0, 100), (500, 400, 100, 0), (500, 500, 0, 0), (600, 0, 0, 400), (600, 0, 100, 300), (600, 0, 200, 200), (600, 0, 300, 100), (600, 0, 400, 0), (600, 100, 0, 300), (600, 100, 100, 200), (600, 100, 200, 100), (600, 100, 300, 0), (600, 200, 0, 200), (600, 200, 100, 100), (600, 200, 200, 0), (600, 300, 0, 100), (600, 300, 100, 0), (600, 400, 0, 0), (700, 0, 0, 300), (700, 0, 100, 200), (700, 0, 200, 100), (700, 0, 300,

0), (700, 100, 0, 200), (700, 100, 100, 100), (700, 100, 200, 0), (700, 200, 0, 100), (700, 200,

100, 0), (700, 300, 0, 0), (800, 0, 0, 200), (800, 0, 100, 100), (800, 0, 200, 0), (800, 100, 0,

100), (800, 100, 100, 0), (800, 200, 0, 0), (900, 0, 0, 100), (900, 0, 100, 0), or (900, 100, 0, 0).

While each of the ratio examples above provides a sum of all four numbers in each ratio to be 1000, it is contemplated that the sum of all four numbers can be less than 1000, e.g., (1,1, 1,0), (5, 1,0,0) and the like.

[0097] Within each of the numerals above includes integers 1 through 99, and in particular 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50,

51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75,

76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99.

Therefore, the number “100”, “200”, and so on can be replaced in the ratios above with any of the integers 1 through 99.

[0098] In addition, any two of the above components in the ratios above can have individual ratios of 1: 1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1: 10, 1: 11, 1: 12, 1: 13, 1: 14, 1: 15, 1: 16, 1: 17, 1: 18, 1: 19, 1:20, 1:21, 1:22, 1:23, 1:24, 1:25, 1:26, 1:27, 1:28, 1:29, 1:30, 1:31,

1:32, 1:33, 1:34, 1:35, 1:36, 1:37, 1:38, 1:39, 1:40, 1:41, 1:42, 1:43, 1:44, 1:45, 1:46, 1:47,

1:48, 1:49, 1:50, 1:51, 1:52, 1:53, 1:54, 1:55, 1:56, 1:57, 1:58, 1:59, 1:60, 1:61, 1:62, 1:63,

1:64, 1:65, 1:66, 1:67, 1:68, 1:69, 1:70, 1:71, 1:72, 1:73, 1:74, 1:75, 1:76, 1:77, 1:78, 1:79,

1:80, 1:81, 1:82, 1:83, 1:84, 1:85, 1:86, 1:87, 1:88, 1:89, 1:90, 1:91, 1:92, 1:93, 1:94, 1:95,

1:96, 1:97, 1:98, 1:99, or 1: 100, and values in between. Moreover, any two of the above components in the ratios above can have individual ratios of 1: 100, 1:200, 1:300, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900, or 1: 1000 and values in between. These specific molar ratios may provide different balances between the two active ingredients, potentially allowing for optimization of the therapeutic efficacy of the composition for different patients or conditions.

[0099] Compounds of (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, and (4) glucagon in the molar ratios described in this disclosure include pharmaceutically acceptable salts, pharmaceutically acceptable stereoisomers, prodrugs, enantiomers, diastereomers, hydrates, co-crystals, and polymorphs thereof.

[0100] In certain instances, the combination includes (1) a PDE-5 inhibitor, (2) a GLP- 1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon present at an amount of greater than about: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 250 mg, or 300 mg. The combination can include (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon present at an amount greater than about: 25 mg, 50 mg, 100 mg, 200 mg, 250 mg, or 300 mg.

[0101] The combination can include (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon present at an amount greater than about: 1 mg to about 10 mg, 1 mg to about 25 mg, 1 mg to about 50 mg, 5 mg to about 10 mg, 5 mg to about 25 mg, 5 mg to about 50 mg, 10 mg to about 25 mg, 10 mg to about 50 mg, 50 mg to about 100 mg, 100 mg to about 200 mg, or 200 mg to about 300 mg.

[0102] The combination can include (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon in an amount of at least about: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 250 mg, or 300 mg. The combination can include (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon present at an amount of at least about: 25 mg, 50 mg, 100 mg, 200 mg, 250 mg, or 300 mg. The combination can include (1) a PDE-5 inhibitor, (2) a GLP- 1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon present at an amount of at least about: 1 mg to about 10 mg, 1 mg to about 25 mg, 1 mg to about 50 mg, 5 mg to about 10 mg, 5 mg to about 25 mg, 5 mg to about 50 mg, 10 mg to about 25 mg, 10 mg to about 50 mg, 50 mg to about 100 mg, 100 mg to about 200 mg, or 200 mg to about 300 mg.

[0103] The combination can include (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon present in an amount of about: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, 200 mg, 250 mg, or 300 mg. The combination can include (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon present at an amount of about: 25 mg, 50 mg, 100 mg, 200 mg, 250 mg, or 300 mg. The combination can include (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon present at an amount of about: 1 mg to about 10 mg, 1 mg to about 25 mg, 1 mg to about 50 mg, 5 mg to about 10 mg, 5 mg to about 25 mg, 5 mg to about 50 mg, 10 mg to about 25 mg, 10 mg to about 50 mg, 50 mg to about 100 mg, 100 mg to about 200 mg, or 200 mg to about 300 mg.

[0104] A PDE-5 inhibitor, a GLP- 1 agonist or DPP-4 inhibitor, a GIP agonist, or glucagon can be present in the combinations described herein relative to the weight of the patient (e.g., mg/kg). In some instances, the (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon is present in an amount equivalent to about: 0.0001 mg/kg to about 200 mg/kg, 0.001 mg/kg to about 200 mg/kg, 0.01 mg/kg to about 200 mg/kg, 0.01 mg/kg to about 150 mg/kg, 0.01 mg/kg to about 100 mg/kg, 0.01 mg/kg to about 50 mg/kg, 0.01 mg/kg to about 25 mg/kg, 0.01 mg/kg to about 10 mg/kg, or 0.01 mg/kg to about 5 mg/kg, 0.05 mg/kg to about 200 mg/kg, 0.05 mg/kg to about 150 mg/kg, 0.05 mg/kg to about 100 mg/kg, 0.05 mg/kg to about 50 mg/kg, 0.05 mg/kg to about 25 mg/kg, 0.05 mg/kg to about 10 mg/kg, or 0.05 mg/kg to about 5 mg/kg, 0.5 mg/kg to about 200 mg/kg, 0.5 mg/kg to about 150 mg/kg, 0.5 mg/kg to about 100 mg/kg, 0.5 mg/kg to about 50 mg/kg, 0.5 mg/kg to about 25 mg/kg, 0.5 mg/kg to about 10 mg/kg, or 0.5 mg/kg to about 5 mg/kg. In other instances the (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon is present in an amount equivalent to about: 1 mg/kg to about 200 mg/kg, 1 mg/kg to about 150 mg/kg, 1 mg/kg to about 100 mg/kg, 1 mg/kg to about 50 mg/kg, 1 mg/kg to about 25 mg/kg, 1 mg/kg to about 10 mg/kg, or 1 mg/kg to about 5 mg/kg.

[0105] In certain instances the therapeutically effective amount of combination hereof is determined as an amount provided in a package insert provided with the combination. The term package insert refers to instructions customarily included in commercial packages of medicaments approved by the FDA or a similar regulatory agency of a country other than the US, which contains information about, for example, the usage, dosage, administration, contraindications, and/or warnings concerning the use of such medicaments.

[0106] At least one of the (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon in the molar ratios described in this disclosure can be provided in amounts that are synergistic with another member of the group. The term synergistic refers to a combination described herein (e.g., (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon - including, optionally, coadministration with another active agent such as an anti-neurodegenerative agent described herein) or a combination of regimens such as those described herein that is more effective than the additive effects of each individual therapy or regimen. [0107] A synergistic effect of a combination described herein can permit the use of lower dosages of one or more of the components of the combination (e.g., (1) a PDE-5 inhibitor, (2) a GLP- 1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon in the molar ratios described in this disclosure). A synergistic effect can permit less frequent administration of at least one of the administered therapies (e.g., (1) a PDE-5 inhibitor, (2) a GLP- 1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon in the molar ratios described in this disclosure) to a subject with a disease, disorder, or condition described herein. Such lower dosages and reduced frequency of administration can reduce the toxicity associated with the administration of at least one of the therapies (e.g., (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon in the molar ratios described in this disclosure) to a subject without reducing the efficacy of the treatment. A synergistic effect as described herein avoid or reduce adverse or unwanted side effects associated with the use of any therapy.

[0108] Pharmaceutical Compositions

[0109] Combinations described herein can be provided as a pharmaceutical composition suitable for administration via any route to a patient described herein including but not limited to: oral, mucosal (e.g., nasal, inhalation, pulmonary, sublingual, vaginal, buccal, or rectal), parenteral (e.g., subcutaneous, intravenous, bolus injection, intramuscular, or intra-arterial), topical (e.g., eye drops or other ophthalmic preparations), transdermal or transcutaneous administration to a patient.

[0110] Exemplary of dosage forms include: tablets; caplets; capsules (e.g., gelatin capsules); cachets; lozenges; suppositories; powders; gels; liquid dosage forms suitable for parenteral administration to a patient; and sterile solids (e.g., crystalline or amorphous solids) that can be reconstituted to provide liquid dosage forms suitable for parenteral administration to a patient.

[0111] Pharmaceutical compositions and dosage forms described herein typically include one or more excipients. Suitable excipients are well known to those skilled in the art of pharmacy. Whether a particular excipient is suitable for incorporation into a pharmaceutical composition or dosage form depends on a variety of factors such as, for example, the intended route of administration to the patient. Pharmaceutical compositions described herein can include other agents such as stabilizers, lubricants, buffers, and disintegrants that can reduce the rate by which an active ingredient can decompose in a particular formulation. [0112] Pharmaceutical compositions described herein can in certain instances include additional active agents other than those in the combinations described herein (e.g., an anti-neurodegenerative agent such as those described herein) in an amount provided herein.

[0113] In one embodiment, one of (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon is provided in an oral dosage form such as a tablet or capsule. In another embodiment, one of (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon is supplied as a powder (e.g., lyophilized powder) that can be resuspended in a liquid suitable for parenteral administration.

[0114] The combinations described herein can be provided in forms convenient to or facilitate their administration to a patient. For example, the combination can be formulated as a tablet, capsule, or as a powder (e.g., lyophilized powder) that can be resuspended in a liquid suitable for parenteral administration.

[0115] Combinations described herein can be provided as controlled release pharmaceutical products, which have a goal of improving drug therapy over that achieved by their non-controlled counterparts. Controlled release formulations can extend activity of the drug, reduce dosage frequency, and increase subject compliance. In addition, controlled release formulations can be used to affect the time of onset of action or other characteristics, such as blood levels of the drug, and can thus affect the occurrence of side (e.g., adverse) effects.

[0116] Methods

[0117] The combinations, pharmaceutical compositions, and kits described herein are useful for treating diseases, disorders, or alleviating, ameliorating, or eliminating the symptoms of diseases and disorders such as, for example, neurodegenerative disorders. It is to be understood that the methods described herein pertain to administration of combinations and pharmaceutical compositions described herein, and such combinations and pharmaceutical compositions can be provided in the form of a kit as described herein. Provided herein are methods of treating neurodegeneration by administering a therapeutically effective amount of a combination described herein to a patient in need thereof. Also provided herein are methods of managing neurodegeneration by administering therapeutically effective amount of a combination described herein to a patient in need thereof.

[0118] In an aspect the methods of treating neurodegeneration provide for methods for reducing amyloid or Ap burden in an individual by administering a therapeutically effective amount of a combination described herein. In some embodiments, neurodegeneration is reduced by at least about 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%.

[0119] The methods of treating neurodegeneration described herein also provide for methods for increasing or otherwise prolonging time to neurodegenerative disorder progression. Time to disease progression can be prolonged in a patient by administering a therapeutically effective amount of a combination described herein. In some embodiments, the increase is a comparison between the time to disease progression without treatment and with treatment with a combination described herein. In some embodiments, the methods described herein prolong the time to disease progression by at least 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 10 years, 15 years, 20 years, 25 years, or more, including values in between.

[0120] The methods of treating neurodegenerative disorders described herein also provide for methods for increasing or otherwise prolonging survival of patients diagnosed with neurodegenerative disorders as described herein. Patient survival can be prolonged by administering a therapeutically effective amount of a combination described herein. In some embodiments, the increase is a comparison between the survival without treatment and with treatment with a combination as described herein. In some embodiments, the methods described herein prolong survival by at least 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 10 years, 15 years, 20 years, 25 years, or more, including values in between.

[0121] The methods of treating neurodegenerative disorders described herein also provide for methods for increasing progression-free survival of patients diagnosed with neurodegenerative disorders as described herein. Patient progression-free survival can be prolonged by administering a therapeutically effective amount of a combination described herein. In some embodiments, the increase is a comparison between the progression-free survival without treatment and with treatment with a combination as described herein. In some embodiments, the methods described herein increase progression-free survival by at least 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 2 years, 3 years, 4 years, 5 years, 10 years, 15 years, 20 years, 25 years, or more, including values in between. [0122] Target Neurodegenerative Disorders

[0123] Synucleinopathies are a group of neurodegenerative disorders characterized by the abnormal accumulation and aggregation of alpha- sy nuclein protein in various parts of the nervous system. In some cases, these disorders may share common pathological mechanisms, but they often present with distinct clinical features. The following outlines several known synucleinopathies:

[0124] Parkinson's Disease (PD): PD may be characterized by the loss of dopaminergic neurons in the substantia nigra pars compacta and the presence of Lewy bodies containing aggregated alpha- synuclein. In some cases, patients may experience motor symptoms such as tremor, rigidity, bradykinesia, and postural instability. Non-motor symptoms may include cognitive impairment, depression, sleep disorders, and autonomic dysfunction.

[0125] Parkinson's Disease Dementia (PDD): PDD may develop in some patients with long-standing PD. It may be characterized by cognitive decline, including impairments in attention, executive function, and visuospatial abilities. In some cases, PDD may be associated with a more widespread distribution of Lewy bodies in cortical and limbic regions.

[0126] Dementia with Lewy Bodies (DLB): DLB may share features with both PD and Alzheimer's disease. It may be characterized by fluctuating cognition, visual hallucinations, and parkinsonism. In some cases, patients may experience rapid eye movement (REM) sleep behavior disorder. The distribution of Lewy bodies in DLB may be more widespread than in PD, often affecting cortical areas.

[0127] Multiple System Atrophy (MSA): MSA may be characterized by a combination of parkinsonian, cerebellar, and autonomic symptoms. In some cases, it may be divided into two subtypes: MSA-P (predominant parkinsonism) and MSA-C (predominant cerebellar ataxia). Alpha- synuclein aggregates in MSA may primarily affect oligodendrocytes, forming glial cytoplasmic inclusions.

[0128] Pure Autonomic Failure (PAF): PAF may be characterized by progressive autonomic dysfunction without significant central nervous system involvement. In some cases, patients may experience orthostatic hypotension, gastrointestinal disturbances, and urogenital dysfunction. Alpha- synuclein aggregates may be found in peripheral autonomic neurons.

[0129] Lewy Body Variant of Alzheimer's Disease (LBV): LBV may represent a condition where patients exhibit pathological features of both Alzheimer's disease and DLB.

In some cases, patients may show cognitive decline typical of Alzheimer's disease along with some features of DLB, such as visual hallucinations or fluctuating cognition.

[0130] In some aspects, these synucleinopathies may share common pathological mechanisms related to alpha- synuclein aggregation and neuronal dysfunction. However, the specific distribution of alpha- synuclein pathology and the affected cell types may vary among these disorders, contributing to their distinct clinical presentations.

[0131] The development of therapeutic strategies targeting alpha- synuclein aggregation, such as the combination of Lamotrigine and Rivastigmine described in this disclosure, may have potential applications across various synucleinopathies. In some cases, such approaches may address the underlying pathological processes common to these disorders, potentially offering new avenues for prevention, alleviation, or treatment of these debilitating neurodegenerative conditions.

[0132] Alzheimer's disease (AD) is a progressive neurodegenerative disorder that may be characterized by cognitive decline, memory loss, and behavioral changes. In some cases, AD may be the most common cause of dementia in older adults. The disease may be associated with the accumulation of Ap plaques and neurofibrillary tangles composed of hyperphosphorylated tau protein in the brain.

[0133] In some aspects, the pathological hallmarks of AD may include:

[0134] 1. Ap plaques: These extracellular deposits may consist of aggregated betaamyloid peptides, which may be derived from the amyloid precursor protein (APP).

[0135] 2. Neurofibrillary tangles: These intracellular aggregates may be composed of hyperphosphorylated tau protein, which may disrupt normal neuronal function.

[0136] 3. Neuronal loss: Progressive degeneration of neurons, particularly in regions such as the hippocampus and cortex, may occur.

[0137] 4. Synaptic dysfunction: Impairment of synaptic transmission and plasticity may contribute to cognitive decline.

[0138] In some cases, AD may progress through several stages, from mild cognitive impairment to severe dementia. The disease may affect various cognitive domains, including memory, language, executive function, and visuospatial abilities.

[0139] Other neurodegenerative diseases related to AD may include:

[0140] Vascular dementia: This form of dementia may be caused by reduced blood flow to the brain, often due to stroke or other vascular issues.

[0141 ] Frontotemporal dementia (FTD): FTD may be characterized by changes in behavior, personality, and language abilities, often with an earlier onset than AD.

[0142] Lewy body dementia (LBD): This condition may share features with both AD and Parkinson's Disease, and may be characterized by cognitive fluctuations, visual hallucinations, and parkinsonism.

[0143] Mixed dementia: In some cases, individuals may exhibit pathological features of multiple types of dementia, such as AD and vascular dementia.

[0144] Posterior cortical atrophy (PCA): This rare form of dementia may primarily affect visual processing and spatial awareness.

[0145] Primary progressive aphasia (PPA): PPA may be characterized by progressive language impairment, which may be a variant of FTD or AD.

[0146] Corticobasal degeneration (CBD): This rare neurological disorder may affect movement and cognition, often presenting with asymmetric motor symptoms and cognitive impairment.

[0147] Progressive supranuclear palsy (PSP): PSP may be characterized by problems with balance, eye movements, and cognitive function.

[0148] Amyloidoses are also target neurodegenerative disorders of the present disclosure. In some aspects, these neurodegenerative disorders may share common pathological mechanisms, such as protein aggregation, mitochondrial dysfunction, and neuroinflammation, including mechanisms involved in synucleinopathies. However, the specific proteins involved, the regions of the brain affected, and the clinical presentations may vary among these disorders.

[0149] The development of therapeutic strategies targeting multiple pathological processes, such as the combination of (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon in the molar ratios described in this disclosure, may have potential applications across various neurodegenerative disorders. In some cases, such approaches may address the underlying pathological processes common to these disorders, potentially offering new avenues for prevention, alleviation, or treatment of these debilitating conditions.

[0150] The combinations described herein can include administration of each therapy (e.g., (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon in the molar ratios described in this disclosure, optionally including another anti-neurodegenerative agent), where the administration is performed simultaneously or sequentially (in either order). In one embodiment, (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, and (4) glucagon in the molar ratios provided herein are administered simultaneously (e.g., within at least 1 to 5 min of each other). In another embodiment, the (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, and (4) glucagon in the molar ratios provided herein are administered sequentially (e.g., within at least 10 min, 15 min, 30 min, 1 h, 2 h, 5 h, 10 h, 12 h, 1 day, 2 days, 5 days, 7 days, 14 days, or 21 days of each other).

[0151] The combinations of the present disclosure can be administered, for example, once a day (QD), twice daily (BID), once a week (QW), twice weekly (BIW), three times a week (TIW), or monthly (QM) regularly on a continuous basis or intermittent basis such as BIW for 3 months then resume a month later. For example, the combinations of the present disclosure can be administered BID. The combinations of the present disclosure can be administered TIW. In certain instances, the combinations of the present disclosure can be administered 2 to 3 times a week. In another embodiment, the combinations of the present disclosure is administered QD. The combinations of the present disclosure can be administered QD for about: 1 day to about 7 days, 1 day to about 14 days, 1 day to about 21 days, 1 day to about 28 days, or daily until disease progression changes or unacceptable toxicity occurs. The administration of combinations of the present disclosure can, in part, depend upon the tolerance of the patient where greater tolerance can allow greater or more frequent administration. Alternatively, where a patient shows poor tolerance to (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon, a lesser amount of one individual compounds or a less frequent dosing of the combination can be performed. Combinations of the present disclosure can be administered in any regimen as described herein.

[0152] For example, combinations of the present disclosure can be administered at an amount of about: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, or 200 mg, QD. For example, combinations of the present disclosure can be administered at an amount of about: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, or 200 mg, BIW. For example, combinations of the present disclosure can be administered at an amount of about: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, or 200 mg, TIW. For example, combinations of the present disclosure can be administered at an amount of about: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, or 200 mg, QW. For example, combinations of the present disclosure can be administered at an amount of about: 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 60 mg, 70 mg, 80 mg, 85 mg, 90 mg, 100 mg, 125 mg, 150 mg, 175 mg, or 200 mg, Q2W. For example, combinations of the present disclosure can be administered at an amount of about 5 mg or about 10 mg, QD. For example, combinations of the present disclosure can be administered at an amount of about 5 mg or about 10 mg, BIW. For example, combinations of the present disclosure can be administered at an amount of about 5 mg or about 10 mg, TIW. For example, combinations of the present disclosure can be administered at an amount of about 5 mg or about 10 mg, QW. For example, combinations of the present disclosure can be administered at an amount of about 5 mg or about 10 mg, Q2W. Administration of combinations of the present disclosure can be continuous. Administration of combinations of the present disclosure can be intermittent.

[0153] For example, combinations of the present disclosure can be administered at an amount of about: 1 mg to about 10 mg, 1 mg to about 25 mg, 1 mg to about 50 mg, 5 mg to about 10 mg, 5 mg to about 25 mg, 5 mg to about 50 mg, 10 mg to about 25 mg, 10 mg to about 50 mg, 50 mg to about 100 mg, or 100 mg to about 200 mg, QD. For example, combinations of the present disclosure can be administered at an amount of about: 1 mg to about 10 mg, 1 mg to about 25 mg, 1 mg to about 50 mg, 5 mg to about 10 mg, 5 mg to about 25 mg, 5 mg to about 50 mg, 10 mg to about 25 mg, 10 mg to about 50 mg, 50 mg to about 100 mg, or 100 mg to about 200 mg, BIW. For example, combinations of the present disclosure can be administered at an amount of about: 1 mg to about 10 mg, 1 mg to about 25 mg, 1 mg to about 50 mg, 5 mg to about 10 mg, 5 mg to about 25 mg, 5 mg to about 50 mg, 10 mg to about 25 mg, 10 mg to about 50 mg, 50 mg to about 100 mg, or 100 mg to about 200 mg, TIW. For example, combinations of the present disclosure can be administered at an amount of about: 1 mg to about 10 mg, 1 mg to about 25 mg, 1 mg to about 50 mg, 5 mg to about 10 mg, 5 mg to about 25 mg, 5 mg to about 50 mg, 10 mg to about 25 mg, 10 mg to about 50 mg, 50 mg to about 100 mg, or 100 mg to about 200 mg, QW. For example, combinations of the present disclosure can be administered at an amount of about: 1 mg to about 10 mg, 1 mg to about 25 mg, 1 mg to about 50 mg, 5 mg to about 10 mg, 5 mg to about 25 mg, 5 mg to about 50 mg, 10 mg to about 25 mg, 10 mg to about 50 mg, 50 mg to about 100 mg, or 100 mg to about 200 mg, Q2W. Administration of combinations of the present disclosure can be continuous. Administration of combinations of the present disclosure can be intermittent.

[0154] For example, combinations of the present disclosure can be administered at an amount of about: 0.0001 mg/kg to about 200 mg/kg, 0.001 mg/kg to about 200 mg/kg, 0.01 mg/kg to about 200 mg/kg, 0.01 mg/kg to about 150 mg/kg, 0.01 mg/kg to about 100 mg/kg, 0.01 mg/kg to about 50 mg/kg, 0.01 mg/kg to about 25 mg/kg, 0.01 mg/kg to about 10 mg/kg, or 0.01 mg/kg to about 5 mg/kg, 0.05 mg/kg to about 200 mg/kg, 0.05 mg/kg to about 150 mg/kg, 0.05 mg/kg to about 100 mg/kg, 0.05 mg/kg to about 50 mg/kg, 0.05 mg/kg to about 25 mg/kg, 0.05 mg/kg to about 10 mg/kg, or 0.05 mg/kg to about 5 mg/kg, 0.5 mg/kg to about 200 mg/kg, 0.5 mg/kg to about 150 mg/kg, 0.5 mg/kg to about 100 mg/kg, 0.5 mg/kg to about 50 mg/kg, 0.5 mg/kg to about 25 mg/kg, 0.5 mg/kg to about 10 mg/kg, or 0.5 mg/kg to about 5 mg/kg, QD. For example, combinations of the present disclosure can be administered at an amount of about: 0.0001 mg/kg to about 200 mg/kg, 0.001 mg/kg to about 200 mg/kg, 0.5 mg/kg to about 200 mg/kg, 0.5 mg/kg to about 150 mg/kg, 0.5 mg/kg to about 100 mg/kg, 0.5 mg/kg to about 50 mg/kg, 0.5 mg/kg to about 25 mg/kg, 0.5 mg/kg to about 10 mg/kg, or 0.5 mg/kg to about 5 mg/kg, BIW. For example, combinations of the present disclosure can be administered at an amount of about: 0.0001 mg/kg to about 200 mg/kg, 0.001 mg/kg to about 200 mg/kg, 0.5 mg/kg to about 200 mg/kg, 0.5 mg/kg to about 150 mg/kg, 0.5 mg/kg to about 100 mg/kg, 0.5 mg/kg to about 50 mg/kg, 0.5 mg/kg to about 25 mg/kg, 0.5 mg/kg to about 10 mg/kg, or 0.5 mg/kg to about 5 mg/kg, TIW. For example, combinations of the present disclosure can be administered at an amount of about: 0.0001 mg/kg to about 200 mg/kg, 0.001 mg/kg to about 200 mg/kg, 0.5 mg/kg to about 200 mg/kg, 0.5 mg/kg to about 150 mg/kg, 0.5 mg/kg to about 100 mg/kg, 0.5 mg/kg to about 50 mg/kg, 0.5 mg/kg to about 25 mg/kg, 0.5 mg/kg to about 10 mg/kg, or 0.5 mg/kg to about 5 mg/kg, QW. For example, combinations of the present disclosure can be administered at an amount of about: 0.0001 mg/kg to about 200 mg/kg, 0.001 mg/kg to about 200 mg/kg, 0.5 mg/kg to about 200 mg/kg, 0.5 mg/kg to about 150 mg/kg, 0.5 mg/kg to about 100 mg/kg, 0.5 mg/kg to about 50 mg/kg, 0.5 mg/kg to about 25 mg/kg, 0.5 mg/kg to about 10 mg/kg, or 0.5 mg/kg to about 5 mg/kg, Q2W. In one example, combinations of the present disclosure can be administered at an amount of about 15 mg/kg to about 75 mg/kg, QD. In another example, combinations of the present disclosure can be administered at an amount of about 20 mg/kg to about 50 mg/kg. In still another example, combinations of the present disclosure can be administered at an amount of about 0.001 mg/kg, 0.01 mg/kg, 0.05 mg/kg, 0.1 mg/kg, 0.5 mg/kg, 1 mg/kg, 2 mg/kg, 3 mg/kg, 4 mg/kg, 5 mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg, 60 mg/kg, 70 mg/kg, 80 mg/kg, 90 mg/kg, 100 mg/kg, 125 mg/kg, 150 mg/kg, 175 mg/kg, or 200 mg/kg. Administration of combinations of the present disclosure can be continuous. Administration of combinations of the present disclosure can be intermittent. [0155] For example, combinations of the present disclosure can be administered at an amount of about: 1 mg/kg to about 200 mg/kg, 1 mg/kg to about 150 mg/kg, 1 mg/kg to about 100 mg/kg, 1 mg/kg to about 50 mg/kg, 1 mg/kg to about 25 mg/kg, 1 mg/kg to about 10 mg/kg, or 1 mg/kg to about 5 mg/kg, QD. For example, combinations of the present disclosure can be administered at an amount of about: 1 mg/kg to about 200 mg/kg, 1 mg/kg to about 150 mg/kg, 1 mg/kg to about 100 mg/kg, 1 mg/kg to about 50 mg/kg, 1 mg/kg to about 25 mg/kg, 1 mg/kg to about 10 mg/kg, or 1 mg/kg to about 5 mg/kg, BIW. For example, combinations of the present disclosure can be administered at an amount of about: 1 mg/kg to about 200 mg/kg, 1 mg/kg to about 150 mg/kg, 1 mg/kg to about 100 mg/kg, 1 mg/kg to about 50 mg/kg, 1 mg/kg to about 25 mg/kg, 1 mg/kg to about 10 mg/kg, or 1 mg/kg to about 5 mg/kg, TIW. For example, combinations of the present disclosure can be administered at an amount of about: 1 mg/kg to about 200 mg/kg, 1 mg/kg to about 150 mg/kg, 1 mg/kg to about 100 mg/kg, 1 mg/kg to about 50 mg/kg, 1 mg/kg to about 25 mg/kg, 1 mg/kg to about 10 mg/kg, or 1 mg/kg to about 5 mg/kg, QW. For example, combinations of the present disclosure can be administered at an amount of about: 1 mg/kg to about 200 mg/kg, 1 mg/kg to about 150 mg/kg, 1 mg/kg to about 100 mg/kg, 1 mg/kg to about 50 mg/kg, 1 mg/kg to about 25 mg/kg, 1 mg/kg to about 10 mg/kg, or 1 mg/kg to about 5 mg/kg, Q2W. In one example, combinations of the present disclosure can be administered at an amount of about 15 mg/kg to about 75 mg/kg, QD. In another example, combinations of the present disclosure can be administered at an amount of about 20 mg/kg to about 50 mg/kg. In still another example, combinations of the present disclosure can be administered at an amount of about 0.001 mg/kg, 0.01 mg/kg, 0.05 mg/kg, 0.1 mg/kg, 0.5 mg/kg, 1 mg/kg, 2 mg/kg, 3 mg/kg, 4 mg/kg, 5 mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg, 60 mg/kg, 70 mg/kg, 80 mg/kg, 90 mg/kg, 100 mg/kg, 125 mg/kg, 150 mg/kg, 175 mg/kg, or 200 mg/kg. Administration of combinations of the present disclosure can be continuous. Administration of combinations of the present disclosure can be intermittent.

[0156] As used herein, the term daily is intended to mean that a therapeutic compound of a combination described herein, such as (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon, is administered once or more than once each day for a period of time. The term continuous is intended to mean that a therapeutic compound of a combination described herein, such as (1) a PDE-5 inhibitor, (2) a GLP- 1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon, is administered daily for an uninterrupted period of at least 10 days to 52 weeks, to multiple years. The term intermittent or intermittently as used herein is intended to mean stopping and starting at either regular or irregular intervals. For example, intermittent administration of a therapeutic compound of a combination described herein, such as (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon, includes administration for one to six days per week (e.g., 2 to 3 times per week or QD), administration in cycles (e.g., daily administration for two to eight consecutive weeks, then a rest period with no administration at least one day), or, for example, administration on alternate days.

[0157] The combinations described herein can be administered in a regimen. The regimen can be structured to provide therapeutically effective amounts of (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, and (4) glucagon in the molar ratios provided herein, optionally including another anti-neurodegenerative agent, over a predetermined period of time (e.g., an administration time). The regimen can be structured to limit or prevent side-effects or undesired complications of each of the components of the combination described herein. The regimen can be structured in a manner that results in increased effect for both therapies of the combination (e.g., synergy). Regimens useful for treating neurodegenerative disorders can include any number of days, months or years of administration which can be repeated as necessary. Administration periods can be broken by a rest period that includes no administration of at least one therapy. For example, a regimen can include administration periods that include 2, 3, 5, 7, 10, 15, 21, 28, or more days. These periods can be repeated. For example, a regimen can include a set number of days as previously described where the regimen is repeated 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or more times.

[0158] Regimens can include a rest period of at least 1, 2, 3, 5, 7, 10, or more days, where at least one therapy is no longer administered to a patient. The rest period can be determined by, for example, monitoring the reaction of the patient to the drug or by measuring the efficacy of the treatment. A rest period can be applicable to a single therapy, such that only one therapy of a combination described herein is discontinued in the rest period but the other therapy(ies) are still administered. Rest periods can be applied to all of the therapies administered to the subject such that the subject receives no therapy for a set period of time during the rest period.

[0159] Regimens described herein for the treatment of neurodegenerative disorders using the combinations described herein can be continued until disease progression is changed or unacceptable toxicity occurs.

[0160] Regimens for administration of combinations described herein include, for example administration one of (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon can be administered BIW, and administration of another member of the group TIW. For example, one of (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon can be administered QD for about 21 days, and another member of the group can be administered Q2W or Q4W. For example, one of (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon can be administered BIW or TIW, and another member of the group can be administered Q2W. In another exemplary regimen, one of (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon can be administered BIW or TIW, and another member of the group can be administered BIW for 2 or 3 weeks. In still another exemplary regimen, one of (1) a PDE-5 inhibitor, (2) a GLP- 1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon can be administered BIW or TIW, and another member of the group can be administered Q4W. In still another exemplary regimen, one of (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon can be administered BIW, and another member of the group can be administered Q2W, Q3W, or Q4W. In certain instances, such regimens include administration of another anti-neurodegenerative agent administered Q2W, Q3W, or Q4W. In yet another exemplary regimen, one of (1) a PDE-5 inhibitor, (2) a GLP-1 agonist or DPP-4 inhibitor, (3) a GIP agonist, or (4) glucagon can be administered TIW, and another member of the group can be administered Q2W, Q3W, or Q4W.

[0161] It should also be appreciated that the combinations described herein for treating neurodegenerative disorders can be coadministered with other active agents other than those present in the combinations described herein (e.g., anti-neurodegenerative agents). Regimens for administration of a combination described herein, including the exemplary regimens set forth above, can be modified as necessary to include administration of such active agents. Administration of such active agents, e.g., anti-neurodegenerative agents, can be performed QD, QW, QM, BID, BIW, TIW, Q2W, Q3W, or Q4W, or in accordance with prescribing information for such anti-neurodegenerative agents as set forth, for example, in a package insert.

[0162] Referring to Table 1 in the Examples below, the effects of combining ARI 001 and semaglutide on intracellular Ap reduction are presented. The table shows data for AR1001 alone, semaglutide alone, and combinations of the two compounds at various concentrations. In some aspects, AR1001 may be tested at concentrations of 0.2 pM and 0.6 pM, while semaglutide may be tested at concentrations of 0.06 pM, 0.14 pM, and 0.28 pM. The data indicates that when used individually, AR1001 and semaglutide may have limited effects on Ap reduction. For instance, 0.2 pM AR1001 alone may result in a 0.547% reduction, while 0.06 pM semaglutide alone may result in a 0.699% reduction.

[0163] However, when AR 1001 and semaglutide are combined, synergistic effects may be observed. In some cases, the combination of 0.2 pM ARI 001 and 0.06 pM semaglutide may result in a 3.573% reduction in intracellular Ap levels. This reduction may be significantly higher than the sum of the individual effects of AR 1001 and semaglutide at these concentrations. The synergistic effect may be quantified by the AB/(A+B) ratio, where values greater than 1 indicate synergy. For the aforementioned combination, the synergistic effect value may be 2.87, suggesting a strong synergistic interaction.

[0164] In some aspects, other concentration combinations may also demonstrate synergistic effects. For example, 0.2 pM AR1001 combined with 0.14 pM semaglutide may result in a 5.957% reduction, with a synergistic effect value of 2.14. Similarly, 0.6 pM AR1001 combined with 0.06 pM semaglutide may lead to a 7.264% reduction, with a synergistic effect value of 2.57.

[0165] The data presented in Table 1 may suggest that the combination of AR1001 and semaglutide may provide enhanced reduction of intracellular Ap levels compared to either compound alone. This synergistic effect may be observed across various concentration ratios of the two compounds. The findings may indicate potential benefits of combining ARI 001 and semaglutide for addressing conditions associated with elevated intracellular Ap levels.

[0166] Referring to Table 2 in the Examples below, the effects of combining ARI 001 and semaglutide on ILip levels in an LPS-induced inflammation model are presented. The table shows data for AR1001 alone, semaglutide alone, and combinations of the two compounds at various concentrations. In some aspects, AR1001 at concentrations of 2 pM and 6 pM may reduce ILip levels by 0.41% and 17.43%, respectively. Semaglutide alone at concentrations of 0.6 pM, 1.4 pM, and 2.8 pM may reduce ILip levels by 9.85%, 32.28%, and 56.66%, respectively.

[0167] When AR 1001 and semaglutide are combined, synergistic effects in reducing ILip levels may be observed. For instance, the combination of 2 pM AR1001 with 0.6 pM semaglutide may result in a 14.52% reduction in ILip levels, with a synergistic effect value of 1.42. In some cases, the combination of 6 pM ARI 001 with 0.6 pM semaglutide may lead to a 56.12% reduction in ILip levels, demonstrating a synergistic effect value of 2.06. [0168] The synergistic effect values for the combinations tested range from 1.42 to 2.06, indicating that the combined effect of AR1001 and semaglutide on ILip reduction may be greater than the sum of their individual effects. In some aspects, the degree of synergy may vary depending on the specific concentrations of AR1001 and semaglutide used in combination.

[0169] These results suggest that combining AR1001 and semaglutide may provide enhanced anti-inflammatory effects compared to either compound alone, as measured by the reduction in ILip levels. The synergistic interactions between ARI 001 and semaglutide may contribute to their potential efficacy in addressing inflammation-related conditions.

[0170] Referring to Table 3 in the Examples below, the effects of combining AR1001 and semaglutide on TNFa levels in an LPS-induced inflammation model are presented. The table shows data for AR1001 alone, semaglutide alone, and combinations of the two compounds at various concentrations. In some aspects, AR1001 at concentrations of 2 pM and 6 pM may reduce TNFa levels by 6.94% and 8.92%, respectively. Semaglutide alone at concentrations of 0.6 pM, 1.4 pM, and 2.8 pM may reduce TNFa levels by 8.15%, 23.57%, and 39.78%, respectively.

[0171] When AR 1001 and semaglutide are combined, synergistic effects in reducing TNFa levels may be observed. For instance, the combination of 2 pM ARI 001 with 0.6 pM semaglutide may result in a 16.17% reduction in TNFa levels, with a synergistic effect value of 1.07. This synergistic effect value, being greater than 1, suggests that the combination may be more effective in reducing TNFa levels than what would be expected from the sum of their individual effects.

[0172] In some cases, increasing the concentration of semaglutide in combination with 2 pM AR1001 may lead to greater reductions in TNFa levels. For example, combining 2 pM AR1001 with 1.4 pM semaglutide may result in a 32.03% reduction, while the combination with 2.8 pM semaglutide may achieve a 52.82% reduction. The synergistic effect values for these combinations may be 1.05 and 1.13, respectively.

[0173] The data also shows that increasing the concentration of ARI 001 to 6 pM in combination with various concentrations of semaglutide may lead to even greater reductions in TNFa levels. For instance, the combination of 6 pM AR1001 with 0.6 pM semaglutide may result in a 19.76% reduction, with a synergistic effect value of 1.16. Combining 6 pM AR1001 with 1.4 pM or 2.8 pM semaglutide may lead to reductions of 35.75% and 56.68%, respectively, with synergistic effect values of 1.10 and 1.16.

[0174] These results suggest that combinations of AR1001 and semaglutide may exhibit synergistic effects in reducing TNFa levels across various concentration ratios. The synergistic effect values ranging from 1.05 to 1.16 indicate that the combinations may be more effective than what would be expected from the individual effects of each compound alone.

[0175] Referring to Table 4 in the Examples below, the effects of combining ARI 001 and Sitagliptin on Ap reduction are presented. The table shows data for various concentrations of AR1001 and Sitagliptin, both individually and in combination, and their impact on Ap levels. In some aspects, AR1001 alone may reduce Ap levels, with a 2.36% reduction observed at 0.2 pM and a 31.47% reduction at 0.6 pM. Sitagliptin alone may also exhibit Ap-reducing effects, with reductions of 0.75% at 1 pM and 9.40% at 2 pM.

[0176] When ARI 001 and Sitagliptin are combined, the Ap reduction effects may be enhanced. For instance, the combination of 0.2 pM AR1001 with 1 pM Sitagliptin may result in a 5.83% reduction in Ap levels. In some cases, increasing the concentration of Sitagliptin to 2 pM while maintaining ARI 001 at 0.2 pM may lead to a 14.52% reduction. The combination of 0.6 pM AR1001 with 1 pM Sitagliptin may produce a 35.83% reduction in Ap levels, while increasing Sitagliptin to 2 pM in this combination may result in a 45.35% reduction.

[0177] The synergistic effects of these combinations are evaluated using the AB/(A+B) ratio. Values greater than 1 for this ratio may indicate synergistic interactions between ARI 001 and Sitagliptin. In some aspects, the combination of 0.2 pM ARI 001 with 1 pM Sitagliptin may exhibit a synergistic effect with an AB/(A+B) ratio of 1.874.

[0178] These results suggest that combining AR1001 and Sitagliptin may lead to enhanced Ap reduction compared to either compound alone. The synergistic effects observed may vary depending on the specific concentrations and ratios of AR1001 and Sitagliptin used. In some cases, the combination of these compounds may provide a more potent approach for reducing Ap levels than using either compound individually.

[0179] In some aspects, the combination of AR 1001 and sitagliptin may exhibit synergistic effects in reducing ILip levels in an LPS-induced inflammation model. Referring to Table 5 in the Examples below, the data shows the effects of AR1001 and sitagliptin, both individually and in combination, on ILip levels.

[0180] ARI 001 alone may reduce ILip levels to varying degrees depending on the concentration used. For example, at 2 pM, AR1001 may reduce ILip levels by 8.07%, while at 6 pM, the reduction may increase to 45.15%. Similarly, sitagliptin alone may also reduce ILip levels, with reductions of 12.23% and 22.62% observed at 10 pM and 20 pM concentrations, respectively.

[0181] When ARlOOl and sitagliptin are combined, the reduction in ILip levels may be greater than the sum of their individual effects. For instance, the combination of 2 pM ARlOOl with 10 pM sitagliptin may result in a 23.54% reduction in ILip levels, with a synergistic effect value of 1.16. This synergistic effect value, being greater than 1, suggests that the combination may be more effective than what would be expected from simply adding the effects of the individual compounds.

[0182] The synergistic effects may be observed across various concentration combinations. For example, 2 pM ARlOOl combined with 20 pM sitagliptin may lead to a 37.69% reduction in ILip levels, with a synergistic effect value of 1.23. Higher concentrations of ARlOOl combined with sitagliptin may result in even greater reductions in ILip levels. The combination of 6 pM ARlOOl with 10 pM sitagliptin may reduce ILip levels by 63.36%, while 6 pM ARlOOl with 20 pM sitagliptin may achieve a 74.80% reduction.

[0183] These results suggest that the combination of ARlOOl and sitagliptin may provide enhanced anti-inflammatory effects compared to either compound alone. The synergistic effect values ranging from 1.10 to 1.23 for the various combinations indicate that this enhanced effect may be consistently observed across different concentration ratios of ARlOOl and sitagliptin.

[0184] Referring to Table 6 of the Examples below, the effects of combining ARlOOl and sitagliptin on TNFa levels in an LPS-induced inflammation model are presented. The table shows data for various concentrations of ARlOOl and sitagliptin, both individually and in combination. In some aspects, ARlOOl alone may reduce TNFa levels, with a 5.08% reduction observed at 2 pM and a 22.95% reduction at 6 pM. Sitagliptin alone may also exhibit TNFa-reducing effects, with a 0.49% reduction at 10 pM and a 16.63% reduction at 20 pM.

[0185] When ARlOOl and sitagliptin are combined, synergistic effects in reducing TNFa levels may be observed. For instance, the combination of 2 pM ARlOOl with 10 pM sitagliptin may result in a 6.64% reduction in TNFa levels, with a synergistic effect value of 1.19. Similarly, 2 pM ARlOOl combined with 20 pM sitagliptin may lead to a 24.34% reduction, with a synergistic effect value of 1.12.

[0186] In some cases, higher concentrations of ARlOOl combined with sitagliptin may produce even greater reductions in TNFa levels. For example, 6 pM ARlOOl combined with 10 pM sitagliptin may result in a 27.84% reduction, with a synergistic effect value of 1.19. The highest reduction of 43.99% may be observed when 6 pM ARI 001 is combined with 20 pM sitagliptin, demonstrating a synergistic effect value of 1.11.

[0187] These results suggest that combinations of AR1001 and sitagliptin may produce synergistic effects in reducing TNFa levels across various concentration ratios. The synergistic effect values greater than 1 for all tested combinations indicate that the combined treatments may be more effective in reducing TNFa levels than what would be expected from the sum of their individual effects.

[0188] Referring to Table 7 of the Example below, the effects of combining ARI 001 and vildagliptin on Ap reduction are presented. In some aspects, the table may show the individual and combined effects of different concentrations of AR 1001 and vildagliptin on Ap levels. The data may indicate that when AR1001 and vildagliptin are administered together, they may produce a greater reduction in Ap levels compared to either compound alone.

[0189] In some cases, AR1001 at concentrations of 0.2 pM and 0.6 pM may reduce Ap levels by 2.36% and 31.47% respectively when administered alone. Vildagliptin, when used individually, may show Ap reduction rates of 3.71% at 1.0 pM and 42.64% at 2.0 pM concentrations.

[0190] The combination of AR 1001 and vildagliptin may exhibit synergistic effects in reducing Ap levels. For instance, 0.2 pM AR1001 combined with 1.0 pM vildagliptin may result in a 7.34% reduction, while 0.2 pM ARI 001 with 2.0 pM vildagliptin may lead to a 45.37% reduction. These combination effects may be greater than the sum of the individual effects of each compound at the corresponding concentrations.

[0191] The synergistic effects may be quantified using the AB/(A+B) ratio, where values greater than 1 may indicate synergism. In some aspects, the combination of 0.2 pM AR1001 and 1.0 pM vildagliptin may show an AB/(A+B) ratio of 1.210, suggesting a synergistic interaction. The highest synergistic effect may be observed for the combination of 0.6 pM AR1001 and 1.0 pM vildagliptin, with an AB/(A+B) ratio of 1.405 and a corresponding Ap reduction rate of 49.43%.

[0192] These results may suggest that combining AR1001 and vildagliptin at certain concentration ratios may lead to enhanced Ap reduction compared to the individual compounds. The synergistic effects observed may vary depending on the specific concentrations and ratios of AR1001 and vildagliptin used in combination.

[0193] In some aspects, the combination of AR 1001 and vildagliptin may exhibit synergistic effects in reducing ILip levels in an LPS-induced inflammation model. Referring to Table 8 of the Examples below, the data presented illustrates the effects of AR1001 and vildagliptin, both individually and in combination, on ILip levels.

[0194] The table shows that ARI 001 alone at concentrations of 2 pM and 6 pM may reduce ILip levels by 9.02% and 45.68%, respectively. Vildagliptin alone at concentrations of 10 pM and 20 pM may reduce ILip levels by 1.44% and 3.82%, respectively.

[0195] When ARI 001 and vildagliptin are combined, the reduction in ILip levels may be significantly enhanced. For example, the combination of 2 pM ARI 001 with 10 pM vildagliptin may result in a 24.97% reduction in ILip levels, with a synergistic effect value of 2.39. Similarly, the combination of 2 pM AR1001 with 20 pM vildagliptin may lead to a 28.93% reduction, with a synergistic effect value of 2.25.

[0196] Higher concentrations of AR 1001 combined with vildagliptin may produce even greater reductions in ILip levels. The combination of 6 pM AR1001 with 10 pM vildagliptin may result in a 55.26% reduction, while 6 pM AR1001 with 20 pM vildagliptin may lead to a 60.32% reduction in ILip levels. These combinations may demonstrate synergistic effect values of 1.17 and 1.22, respectively.

[0197] The synergistic effect values, calculated as AB/(A+B), may indicate the degree of synergy between AR 1001 and vildagliptin. Values greater than 1 may suggest a synergistic interaction, with higher values potentially indicating stronger synergistic effects. In this case, all tested combinations of AR 1001 and vildagliptin may exhibit synergistic effects in reducing ILip levels, with the most pronounced synergy observed at lower AR1001 concentrations.

[0198] These results may suggest that the combination of AR1001 and vildagliptin could potentially provide enhanced anti-inflammatory effects compared to either compound alone, particularly in reducing ILip levels in LPS-induced inflammation models.

[0199] In some aspects, the combination of AR 1001 and vildagliptin may exhibit synergistic effects in reducing TNFa levels in an LPS-induced inflammation model. Referring to Table 9 of the Examples below, the data presents the results of experiments evaluating the effects of AR 1001 and vildagliptin, both individually and in combination, on TNFa levels.

[0200] The table shows that ARI 001 alone at concentrations of 2 pM and 6 pM may reduce TNFa levels by 5.12% and 23.72%, respectively. Vildagliptin alone at concentrations of 10 pM and 20 pM may reduce TNFa levels by 4.48% and 9.71%, respectively. [0201] When AR1001 and vildagliptin are combined, the reduction in TNFa levels may be greater than the sum of their individual effects. For example, the combination of 2 pM ARlOOl and 10 pM vildagliptin may result in an 11.19% reduction in TNFa levels, with a synergistic effect value (AB/(A+B)) of 1.17. Similarly, the combination of 2 pM ARlOOl and 20 pM vildagliptin may lead to a 16.88% reduction, with a synergistic effect value of 1.14.

[0202] Higher concentrations of ARlOOl combined with vildagliptin may produce even greater reductions in TNFa levels. The combination of 6 pM ARlOOl and 10 pM vildagliptin may result in a 33.80% reduction, with a synergistic effect value of 1.20. The highest reduction of 37.48% may be observed with the combination of 6 pM ARlOOl and 20 pM vildagliptin, demonstrating a synergistic effect value of 1.12.

[0203] In some cases, the synergistic effect values greater than 1 for all tested combinations may indicate that ARlOOl and vildagliptin work synergistically to reduce TNFa levels in this inflammation model. The degree of synergy may vary depending on the specific concentrations and ratios of ARlOOl and vildagliptin used.

[0204] Referring to Table 10 of the Examples below, the effects of combining ARlOOl and linagliptin on Ap reduction are presented. The table shows data for various concentrations of ARlOOl and linagliptin, both individually and in combination. In some aspects, ARlOOl may be tested at concentrations of 0.2 pM and 0.6 pM, while linagliptin may be tested at a concentration of 0.1 pM.

[0205] The data indicates that when used alone, ARlOOl at 0.2 pM and 0.6 pM may result in Ap reduction rates of 2.36% and 31.47%, respectively. Linagliptin alone at 0.1 pM may produce Ap reduction rates of 6.29%.

[0206] In some cases, when ARlOOl and linagliptin are combined, the Ap reduction rates may be significantly higher than those observed with either compound alone. For example, the combination of 0.2 pM ARlOOl and 0.1 pM linagliptin may result in a 15.73% reduction in Ap levels.

[0207] The synergistic effects of these combinations may be evaluated using the AB/(A+B) ratio. Values greater than 1 for this ratio may indicate synergistic interactions between ARlOOl and linagliptin. The combination of 0.2 pM ARlOOl and 0.1 pM linagliptin may exhibit an AB/(A+B) ratio of 1.819, indicating a strong synergistic interaction.

[0208] An Ap reduction rate of 46.50% may be observed for the combination of 0.6 pM ARlOOl and 0.1 pM linagliptin, with an AB/(A+B) ratio of 1.231. This combination may demonstrate a substantial improvement in Ap reduction compared to either compound used individually.

[0209] These results may suggest that the combination of AR1001 and linagliptin could potentially enhance the therapeutic effects in reducing Ap levels, which may be relevant for conditions associated with Ap accumulation. The synergistic interactions observed at various concentration combinations may provide flexibility in dosing strategies for potential therapeutic applications.

[0210] Table 11 of the Examples below presents data on the effects of combining AR1001 and linagliptin on IL-ip levels in an LPS-induced inflammation model. In some aspects, AR1001 and linagliptin may be tested individually and in combination at various concentrations to assess their impact on IL-ip production. The table may show IL-ip fold change values and reduction percentages for different treatment conditions.

[0211] In some cases, ARI 001 alone at concentrations of 2 pM and 6 pM may reduce IL-ip levels by 6.47% and 43.45%, respectively. Linagliptin alone at 1 pM may reduce IL-ip levels by 7.21%. When AR1001 and linagliptin are combined, the reduction in IL-ip levels may be greater than either compound alone. For example, the combination of 2 pM ARI 001 and 1 pM linagliptin may result in a 15.62% reduction in IL-ip levels, while the combination of 6 pM ARI 001 and 1 pM linagliptin may lead to a 60.50% reduction.

[0212] The synergistic effect of combining AR 1001 and linagliptin may be evaluated using the AB/(A+B) ratio. In some aspects, values greater than 1 for this ratio may indicate a synergistic interaction between the compounds. The data may show AB/(A+B) values of 1.14 and 1.19 for the tested combinations, suggesting synergistic effects in reducing IL-ip levels.

[0213] Referring to Table 12 of the Examples below, the effects of combining AR1001 and linagliptin on TNFa levels in an LPS-induced inflammation model are presented. The table shows data for various concentrations of AR1001 and linagliptin, both individually and in combination. In some aspects, AR1001 alone may reduce TNFa levels in a concentration-dependent manner. For example, 2 pM AR1001 may reduce TNFa levels by 4.77%, while 6 pM AR1001 may reduce TNFa levels by 22.31%. In some cases, linagliptin alone at 1 pM may reduce TNFa levels by 11.33%.

[0214] The combination of AR 1001 and linagliptin may exhibit synergistic effects in reducing TNFa levels. For instance, when 2 pM AR1001 is combined with 1 pM linagliptin, the TNFa reduction may increase to 18.91%, which is higher than the sum of their individual effects. This synergistic effect is quantified by the AB/(A+B) ratio of 1.17, where values greater than 1 indicate synergy. In some aspects, a more pronounced synergistic effect may be observed when 6 pM ARI 001 is combined with 1 pM linagliptin, resulting in a 40.58% reduction in TNFa levels and an AB/(A+B) ratio of 1.21.

[0215] The data in Table 12 may suggest that the combination of AR 1001 and linagliptin can produce greater reductions in TNFa levels compared to either compound alone across different concentration combinations. This synergistic effect may be particularly evident at higher concentrations of AR 1001 combined with linagliptin.

[0216] Referring to FIG. 1, a bar graph is shown illustrating the effects of AR1001 and semaglutide on IL-6 level reduction in an LPS-induced inflammation model. The y-axis of the graph represents IL-6 level reduction as a percentage, ranging from 0 to 100%. The x- axis displays different treatment conditions, including LPS alone, varying concentrations of AR1001 (0.1, 0.5, 1, and 5 pM), and combinations of AR1001 with semaglutide (0.01, 0.1, 0.1, and 0.1 pM respectively).

[0217] In some aspects, the graph may demonstrate that AR 1001 or semaglutide alone have minimal effects on IL-6 reduction when compared to the LPS-only control. However, combinations of AR 1001 and semaglutide may result in significantly greater reductions in IL-6 levels.

[0218] The data suggests that in certain cases, the combination of AR1001 and semaglutide may produce a synergistic effect in reducing IL-6 levels. For example, the combination of 0.5 pM ARI 001 and 0.1 pM semaglutide may show the highest reduction in IL-6 levels among the tested combinations. This reduction may be substantially greater than the sum of the reductions observed for each compound individually at these concentrations.

[0219] In some embodiments, the synergistic effect may be observed across various concentration combinations of AR 1001 and semaglutide. The degree of synergy may vary depending on the specific concentrations used. For instance, the combination of 0.1 pM ARI 001 and 0.1 pM semaglutide may exhibit a different level of synergy compared to the combination of 5 pM AR1001 and 0.1 pM semaglutide.

[0220] The graph includes error bars for each condition, which may indicate the variability of the measurements across different samples or replicates. These error bars may provide information about the reliability and consistency of the observed effects.

[0221] In some cases, the synergistic effects observed in this LPS-induced inflammation model may suggest potential therapeutic applications for the combination of AR1001 and semaglutide in conditions associated with elevated IL-6 levels. However, further studies may be necessary to fully characterize the nature and extent of this synergistic interaction across a broader range of concentrations and in different experimental models. [0222] Table 13 of the Examples below presents data on the effects of combining AR1001 and semaglutide on IL-6 levels in an LPS-induced inflammation model. The table shows various concentrations of ARI 001 (0.1, 0.5, 1, and 5 pM) and semaglutide (0.1 pM) tested individually and in combination.

[0223] In some aspects, AR1001 alone may exhibit modest effects on IL-6 reduction, with reduction percentages ranging from 8.70% to 11.48% across the tested concentrations. Semaglutide alone at 0.1 pM may show minimal IL-6 reduction of 0.99%.

[0224] The combination of AR 1001 and semaglutide may demonstrate synergistic effects in reducing IL-6 levels. For instance, the combination of 0.1 pM AR1001 and 0.1 pM semaglutide may result in a 16.67% reduction in IL-6 levels, with a synergistic effect value of 1.65. This synergistic effect value, being greater than 1, suggests that the combination may be more effective than the sum of the individual effects of AR 1001 and semaglutide.

[0225] In some cases, the synergistic effect may vary depending on the concentration ratio of AR1001 to semaglutide. The data indicates that ratios ranging from 1: 1 to 50: 1 (AR 1001: semaglutide) may all exhibit synergistic effects, with synergistic effect values ranging from 1.16 to 1.65.

[0226] The highest IL-6 reduction of 18.61% may be observed for the combination of 0.5 pM AR1001 and 0.1 pM semaglutide, corresponding to a 5: 1 ratio. This combination may demonstrate a synergistic effect value of 1.49.

[0227] These results suggest that combining AR1001 and semaglutide may provide enhanced anti-inflammatory effects compared to either compound alone in this LPS-induced inflammation model. The synergistic interactions observed across various concentration ratios may indicate flexibility in potential dosing strategies for combined use of these compounds.

[0228] Referring to FIG. 2, a bar graph is shown depicting the effects of AR1001 and semaglutide on IL-6 level reduction in an LPS-induced inflammation model. The y-axis of the graph represents the IL-6 level reduction percentage, ranging from 0 to 100%. The x- axis displays different treatment conditions, including LPS alone, AR1001 at concentrations of 10 and 20 pM, semaglutide at concentrations of 0.1 and 1 pM, and combinations of AR1001 and semaglutide at various concentrations.

[0229] In some aspects, the graph demonstrates that the combination of AR 1001 and semaglutide may result in a greater reduction of IL-6 levels compared to either compound alone. For example, when AR1001 at 10 pM is combined with semaglutide at 0.1 pM, the IL-6 level reduction may be approximately 30%, which may be higher than the reduction observed with either AR1001 or semaglutide alone at these concentrations.

[0230] The data suggests that increasing the concentration of both compounds may lead to enhanced IL-6 reduction. In some cases, the combination of 20 pM ARI 001 and 1 pM semaglutide may produce the highest reduction in IL-6 levels, reaching approximately 55% reduction. This reduction may be substantially greater than the reductions observed with either compound administered individually at these concentrations.

[0231] The graph includes error bars for each condition, which may indicate the variability of the measurements across different samples or replicates. These error bars may provide information about the consistency and reliability of the observed effects.

[0232] In some aspects, the synergistic effects of combining AR 1001 and semaglutide may be particularly pronounced at higher concentrations. For instance, while AR1001 at 20 pM alone may reduce IL-6 levels by approximately 45%, and semaglutide at 1 pM alone may have minimal effect, their combination may result in a reduction of about 55%, suggesting a potential synergistic interaction.

[0233] The data presented in FIG. 2 may support the concept that combining AR 1001 and semaglutide at certain concentrations may provide enhanced anti-inflammatory effects in this LPS-induced inflammation model, as measured by IL-6 level reduction. This combination approach may offer potential benefits in addressing inflammatory conditions where IL-6 plays a significant role.

[0234] Referring to Table 14 of the Examples below, the data illustrates the effects of combining AR1001 and semaglutide on IL-6 levels in an LPS-induced inflammation model. The table presents results for various concentrations of AR1001 (10 pM and 20 pM) and semaglutide (0.1 pM and 1 pM), both individually and in combination.

[0235] In some aspects, ARI 001 alone at 10 pM and 20 pM may reduce IL-6 levels by 25.75% and 46.60% respectively. Semaglutide alone at 0.1 pM and 1 pM may show increases in IL-6 levels of 19.95% and 2.26% respectively.

[0236] When AR1001 and semaglutide are combined, greater reductions in IL-6 levels may be observed. For example, the combination of 10 pM ARI 001 with 0.1 pM semaglutide may result in a 31.33% reduction in IL-6 levels. In some cases, increasing the concentration of semaglutide to 1 pM while maintaining AR1001 at 10 pM may lead to a 36.31% reduction.

[0237] The data suggests that higher concentrations of both compounds may produce more pronounced effects. For instance, the combination of 20 pM AR1001 with 0.1 pM semaglutide may reduce IL-6 levels by 49.16%. The most substantial reduction of 55.68% may be observed with 20 pM AR1001 and 1 pM semaglutide.

[0238] The synergistic effect column in Table 14 displays values greater than 1 for all combinations, which may indicate synergistic interactions between AR 1001 and semaglutide in reducing IL-6 levels. The highest synergistic effect value of 5.40 may be observed for the combination of 10 pM AR1001 and 0.1 pM semaglutide.

[0239] The rate column in Table 14 shows the ratio of AR 1001 to semaglutide concentrations used in each combination treatment. These ratios may range from 10:1 to 200: 1, allowing for analysis of different proportional combinations and their effects on IL-6 reduction.

[0240] Referring to FIG. 3, a bar graph is shown illustrating the effects of AR1001 and Tirzepatide on IL-6 level reduction in an LPS-induced inflammation model. The y-axis of the graph represents the IL-6 level reduction percentage, ranging from 0 to 100%. The x- axis displays different treatment conditions, including LPS alone, AR1001 at concentrations of 10 pM and 20 pM, Tirzepatide at concentrations of 0.1 pM and 1 pM, and combinations of AR1001 and Tirzepatide at various concentrations.

[0241] In some aspects, the graph may demonstrate that combinations of AR 1001 and Tirzepatide result in greater reductions of IL-6 levels compared to either compound administered alone. For instance, when ARI 001 at 10 pM is combined with Tirzepatide at 0.1 pM, the IL-6 level reduction may be significantly higher than the reduction observed with either AR1001 or Tirzepatide alone at these concentrations.

[0242] The highest IL-6 reduction may be observed for the combination of 20 pM ARI 001 and 1 pM Tirzepatide, reaching nearly 90% reduction. This combination may exhibit a synergistic effect, as the reduction in IL-6 levels appears to be greater than the sum of the reductions achieved by each compound individually.

[0243] In some cases, the graph may include error bars for each condition, indicating the variability of the measurements across different samples or replicates. Asterisks above some bars may denote statistical significance, potentially highlighting the combinations that show significantly greater IL-6 reduction compared to control conditions or individual treatments.

[0244] The data presented in FIG. 3 may suggest a dose-dependent effect of both AR1001 and Tirzepatide on IL-6 reduction. Additionally, the graph may indicate potential synergistic action between AR1001 and Tirzepatide in reducing IL-6 levels in this LPS- induced inflammation model.

[0245] Referring to Table 15 of the Examples below, the effects of combining AR1001 and Tirzepatide on IL-6 levels in an LPS-induced inflammation model are presented. The table shows data for various concentrations of ARI 001 (10 pM and 20 pM) and Tirzepatide (0.1 pM and 1 pM), both individually and in combination. In some aspects, AR1001 alone at 10 pM may reduce IL-6 levels by 21.31%, while at 20 pM it may reduce IL- 6 levels by 59.12%. Tirzepatide alone at 0.1 pM and 1 pM may increase IL-6 levels by 18.94% and 2.49%, respectively.

[0246] When AR 1001 and Tirzepatide are combined, synergistic effects in reducing IL-6 levels may be observed. For instance, the combination of 10 pM AR1001 and 0.1 pM Tirzepatide may result in a 59.12% reduction in IL-6 levels, with a synergistic effect value of 24.90. This synergistic effect value, calculated as (AB/A+B), indicates a significantly enhanced reduction compared to the sum of the individual effects of AR1001 and Tirzepatide alone.

[0247] In some cases, the combination of 10 pM ARI 001 and 1 pM Tirzepatide may lead to a 43.45% reduction in IL-6 levels, with a synergistic effect value of 2.31. The combination of 20 pM AR1001 and 0.1 pM Tirzepatide may result in a 51.87% reduction, with a synergistic effect value of 1.29. The highest reduction in IL-6 levels (89.63%) may be observed with the combination of 20 pM ARI 001 and 1 pM Tirzepatide, corresponding to a synergistic effect value of 1.58.

[0248] These results suggest that various combinations of AR1001 and Tirzepatide may produce synergistic effects in reducing IL-6 levels in this inflammation model, with the degree of synergy varying based on the specific concentrations and ratios used. The concentration ratios of AR1001 to Tirzepatide in these combinations range from 10: 1 to 200: 1, providing insights into potential optimal ratios for enhanced anti-inflammatory effects.

[0249] Referring to FIG. 4, a bar graph is shown illustrating the effects of AR1001 and dulaglutide on IL-6 level reduction in an LPS-induced inflammation model. The graph displays IL-6 reduction percentages for various concentrations of AR1001 and dulaglutide, both individually and in combination. In some aspects, the y-axis represents IL-6 reduction as a percentage, ranging from 0 to 100%, while the x-axis shows different treatment conditions.

[0250] The graph may include error bars for each condition, indicating variability in the measurements. In some cases, certain bars are marked with asterisks, which may denote statistical significance levels. The overall trend suggests that combinations of AR1001 and dulaglutide may reduce IL-6 levels to a greater extent compared to either compound administered alone.

[0251] In some aspects, the graph demonstrates that AR 1001 at concentrations ranging from 0.1 to 5 pM, when combined with dulaglutide at concentrations of 0.001 to 0.1 pM, may produce enhanced IL-6 reduction compared to the individual treatments. The highest reduction in IL-6 levels may be observed for specific combinations of AR1001 and dulaglutide, potentially reaching approximately 20-25% reduction.

[0252] The graph may also include a control condition and treatments with LPS alone at 100 ng/ml, providing a baseline for comparison. In some cases, the synergistic effects of AR 1001 and dulaglutide combinations may be particularly evident when compared to these control conditions.

[0253] The data presented in FIG. 4 may suggest that AR1001 and dulaglutide, when administered in combination, could potentially offer enhanced anti-inflammatory effects in conditions associated with elevated IL-6 levels. However, the specific concentrations and ratios of AR1001 and dulaglutide that produce optimal IL-6 reduction may vary and may require further investigation to determine the most effective treatment regimens.

[0254] Referring to Table 16 of the Examples below, the effects of combining AR1001 and dulaglutide on IL-6 levels in an LPS-induced inflammation model are presented. The table shows data for various concentrations of ARI 001 (0.1 to 5 pM) and dulaglutide (0.001 to 0.1 pM), both individually and in combination. In some aspects, AR1001 alone may have limited effects on IL-6 reduction, with the highest reduction of 8.21% observed at 1 pM concentration. Similarly, dulaglutide alone may show modest IL-6 reductions, with a maximum of 7.82% at 0.1 pM concentration.

[0255] The combination of AR 1001 and dulaglutide may result in enhanced IL- 6 reduction compared to either compound alone. For instance, the combination of 0.1 pM AR1001 and 0.01 pM dulaglutide may lead to a 9.54% reduction in IL-6 levels, which is greater than the sum of their individual effects. This synergistic effect is quantified by the AB/(A+B) value of 1.53, where values greater than 1 indicate synergy.

[0256] In some cases, the synergistic effects may be more pronounced at certain concentration ratios. For example, the combination of 0.5 pM AR1001 and 0.01 pM dulaglutide (50:1 ratio) may result in a 26.63% reduction in IL-6 levels, with a synergistic effect value of 1.85. The highest synergistic effect (2.03) may be observed for the combination of 0.1 pM AR1001 and 0.001 pM dulaglutide (100: 1 ratio), resulting in a 4.85% IL-6 reduction.

[0257] The data suggests that the synergistic effects may vary depending on the concentration ratios of AR1001 and dulaglutide. Ratios ranging from 1: 1 to 1000: 1 may exhibit synergistic interactions, with AB/(A+B) values consistently above 1. In some aspects, the most effective combinations for IL-6 reduction may be observed at ratios between 50: 1 and 500: 1.

[0258] These results may indicate that combining AR1001 and dulaglutide at specific concentration ratios could potentially enhance their anti-inflammatory effects in this LPS-induced inflammation model. The synergistic interactions observed may suggest a possible benefit in co-administering these compounds for reducing IL-6 levels, which may have implications for managing inflammatory conditions.

[0259] Referring to FIG. 5, a bar graph is shown depicting the effects of AR 1001 and semaglutide on IL-6 level reduction in a TNF a- induced inflammation model. The y-axis of the graph may represent the IL-6 level reduction percentage, ranging from 0 to 100%. The x-axis may display different treatment conditions, including TNFa alone, ARI 001 alone at concentrations of 10 and 20 pM, semaglutide alone at concentrations of 0.01, 0.1, and 1 pM, and combinations of AR1001 and semaglutide at various concentrations.

[0260] In some aspects, the graph may demonstrate that combinations of AR1001 and semaglutide result in greater IL-6 level reductions compared to either compound alone. For example, while AR1001 at 10 pM or 20 pM alone may show minimal effects on IL-6 reduction, and semaglutide alone at 0.01, 0.1, or 1 pM may also have limited impact, the combinations of these compounds may exhibit substantially higher reductions in IL-6 levels.

[0261] The highest reduction in IL-6 levels may be observed for the combination of 20 pM ARI 001 and 1 pM semaglutide. This combination may demonstrate a synergistic effect, where the reduction in IL-6 levels is greater than the sum of the reductions achieved by each compound individually.

[0262] Error bars included for each data point may indicate variability in the measurements across different samples or replicates. The presence of these error bars may suggest that multiple experiments were conducted to ensure the reliability of the results.

[0263] In some cases, statistical significance symbols may be included above certain bars in the graph. These symbols may indicate that the differences observed between certain treatment conditions are statistically significant, providing further support for the synergistic effects of combining AR 1001 and semaglutide.

[0264] The data presented in FIG. 5 may suggest that the combination of AR 1001 and semaglutide may have potential therapeutic applications in conditions characterized by elevated IL-6 levels or TNFa-induced inflammation. However, it is important to note that these results are based on a specific experimental model, and further studies may be necessary to fully elucidate the mechanisms underlying these synergistic effects and to determine their relevance in other experimental systems or clinical settings.

[0265] Referring to Table 17, the effects of combining AR 1001 and semaglutide on IL-6 levels in a TNFa-induced inflammation model are presented. The table shows data for various concentrations of AR1001 (10 pM and 20 pM) and semaglutide (0.01 pM, 0.1 pM, and 1 pM) alone and in combination.

[0266] In some aspects, ARI 001 alone at 10 pM may increase IL-6 levels by 7.06%, while at 20 pM it may reduce IL-6 levels by 10.41%. Semaglutide alone may increase IL-6 levels at lower concentrations (14.28% increase at 0.01 pM and 13.99% increase at 0.1 pM) but may show a smaller increase (3.26%) at 1 pM.

[0267] The combination of AR 1001 and semaglutide may exhibit synergistic effects in reducing IL-6 levels. For instance, 10 pM AR1001 combined with 0.01 pM semaglutide may reduce IL-6 levels by 23.70%, with a synergistic effect value of 1.11. The combination of 10 pM AR1001 and 0.1 pM semaglutide may result in a 25.34% reduction, with a synergistic effect of 1.20.

[0268] In some cases, higher concentrations of AR 1001 combined with semaglutide may lead to more pronounced synergistic effects. The combination of 20 pM ARI 001 and 0.1 pM semaglutide may reduce IL-6 levels by 24.68%, with a synergistic effect value of 6.89. The highest reduction in IL-6 levels (41.58%) may be observed with 20 pM AR1001 and 1 pM semaglutide, demonstrating a synergistic effect value of 5.81.

[0269] The synergistic effect values greater than 1 for all combinations tested suggest that AR 1001 and semaglutide may work synergistically to reduce IL-6 levels in this TNFa-induced inflammation model. The degree of synergy may vary depending on the specific concentrations and ratios of AR1001 and semaglutide used.

[0270] Referring to FIG. 6, a bar graph is shown depicting the effects of AR1001 and Tirzepatide on IL-6 level reduction in a TNFa-induced inflammation model. The graph displays IL-6 level reduction percentages on the y-axis, ranging from 0 to 100%. The x-axis shows different treatment conditions, including varying concentrations of AR1001 (0.1 to 20 pM) and Tirzepatide (0.001 to 1 pM), both alone and in combination.

[0271] In some aspects, the graph may demonstrate that combinations of AR 1001 and Tirzepatide result in greater IL-6 level reductions compared to either compound administered individually. For instance, the combination of 20 pM AR1001 and 1 pM Tirzepatide may exhibit the most significant reduction in IL-6 levels, as indicated by the tallest bar on the graph. [0272] The graph may include error bars for each condition, which may represent the variability in the measurements across different samples or replicates. In some cases, asterisks above certain bars may denote statistical significance, potentially indicating that the observed differences between treatment conditions are unlikely to have occurred by chance.

[0273] AR 1001 alone may show a concentration-dependent effect on IL- 6 level reduction, with higher concentrations generally resulting in greater reductions. Similarly, Tirzepatide administered individually may also demonstrate some ability to reduce IL-6 levels, though potentially to a lesser extent than AR1001 at the concentrations tested.

[0274] The synergistic effects of combining AR1001 and Tirzepatide may be particularly evident at certain concentration ratios. For example, the combinations of 10 pM ARI 001 with 1 pM Tirzepatide, and 20 pM ARI 001 with 0.1 or 1 pM Tirzepatide, may show substantially greater IL-6 reductions than what might be expected from the sum of their individual effects.

[0275] In some aspects, this data may suggest that the combination of AR 1001 and Tirzepatide may provide enhanced anti-inflammatory effects in TNF a- induced inflammation models compared to either compound alone. The synergistic interaction between these compounds may potentially lead to more effective reduction of inflammatory markers such as IL-6.

[0276] Referring to Table 18 of the Examples below, the effects of combining AR1001 and Tirzepatide on IL-6 levels in a TNFa-induced inflammation model are presented. The table shows data for various concentrations of ARI 001 (ranging from 0.1 pM to 20 pM) and Tirzepatide (ranging from 0.001 pM to 1 pM) alone and in combination. In some aspects, AR1001 alone may exhibit modest reductions in IL-6 levels, with a 6.48% increase at 10 pM and a 7.60% reduction at 20 pM. Tirzepatide alone may show variable effects, with increases in IL-6 levels at lower concentrations and a slight reduction of 0.75% at 0.1 pM.

[0277] When AR 1001 and Tirzepatide are combined, synergistic effects in reducing IL-6 levels may be observed. For instance, the combination of 10 pM AR1001 and 1 pM Tirzepatide may result in a 19.26% reduction in IL-6 levels, with a synergistic effect value of 1.11.

[0278] The data suggests that higher concentrations of both compounds may produce more pronounced synergistic effects. For example, the combination of 20 pM AR1001 and 1 pM Tirzepatide may result in a 36.01% reduction in IL-6 levels, with a notably high synergistic effect value of 10.86. This combination may demonstrate the potential for enhanced anti-inflammatory effects when AR1001 and Tirzepatide are used together at specific concentration ratios.

[0279] In some aspects, the synergistic effects may vary depending on the concentration ratios of AR1001 to Tirzepatide. The table presents ratios ranging from 5: 1 to 200: 1, with each combination showing synergistic effects to varying degrees. This variability in synergistic effects across different concentration ratios may suggest the importance of optimizing the relative concentrations of AR1001 and Tirzepatide for maximal antiinflammatory efficacy in this model.

[0280] Referring to FIG. 7, a bar graph is shown illustrating the effects of AR1001 and dulaglutide on IL-6 levels in a TNFa-induced inflammation model. The y-axis of the graph represents IL-6 concentration in pg/ml, ranging from 0 to 100. The x-axis displays different treatment conditions, including varying concentrations of TNFa, AR1001, and dulaglutide. In some aspects, the graph may demonstrate the potential synergistic effects of combining AR1001 and dulaglutide in reducing IL-6 levels compared to either compound alone.

[0281] The graph may include multiple bars for each treatment condition, with error bars indicating variability in the measurements. In some cases, asterisks above certain bars may denote statistical significance levels for the observed effects. The legend below the graph may provide details on the concentrations of TNFa (5ng/ml), AR1001 (0.1 to 1 pM), and dulaglutide (0.001 to 1 pM) used in each condition.

[0282] In some aspects, the overall trend of the data may suggest that certain combinations of AR1001 and dulaglutide reduce IL-6 levels more effectively than TNFa treatment alone or individual treatments with either compound. For example, the combination of 0.1 pM ARI 001 and 0.01 pM dulaglutide may result in a lower IL-6 concentration compared to either compound administered individually at those concentrations.

[0283] The synergistic effects of AR1001 and dulaglutide may be observed across various concentration combinations. In some cases, the most pronounced reduction in IL-6 levels may be achieved with specific ratios of AR1001 to dulaglutide. For instance, the combination of 0.5 pM AR1001 and 0.01 pM dulaglutide may exhibit a particularly strong synergistic effect in reducing IL-6 levels.

[0284] It is important to note that the effectiveness of the combination therapy may vary depending on the specific concentrations of AR1001 and dulaglutide used. In some aspects, certain concentration ratios may produce more optimal results than others in terms of IL-6 reduction. The data presented in FIG. 7 may provide insights into potential dosing strategies for combining AR 1001 and dulaglutide to achieve maximal anti-inflammatory effects in TNF a- induced inflammation models.

[0285] Referring to Table 19 of the Examples below, the effects of combining AR1001 and dulaglutide on IL-6 levels in a TNFa-induced inflammation model are presented. The table shows data for various concentrations of ARI 001 (0.1 pM to 1 pM) and dulaglutide (0.001 pM to 0.1 pM), both individually and in combination. In some aspects, AR1001 alone may exhibit modest reductions in IL-6 levels, with a 2.53% reduction observed at 1 pM concentration. Dulaglutide alone may also show some IL-6 reduction, with a maximum of 2.97% reduction at 0.01 pM concentration.

[0286] When AR1001 and dulaglutide are combined, the IL-6 reduction may be enhanced compared to either compound alone. For instance, the combination of 0.1 pM AR1001 and 0.01 pM dulaglutide may result in a 19% reduction in IL-6 levels. This combination may demonstrate a synergistic effect, as indicated by the AB/(A+B) value of 6.44, which is greater than 1.

[0287] In some cases, other concentration ratios of AR1001 and dulaglutide may also exhibit synergistic effects. For example, the combination of 0.5 pM AR1001 and 0.1 pM dulaglutide may lead to a 17.74% reduction in IL-6 levels, with an AB/(A+B) value of 4.64. The synergistic effects may be observed across various concentration ratios, ranging from 1: 1 to 500: 1 (AR 1001: dulaglutide).

[0288] The data in Table 19 may suggest that combining AR 1001 and dulaglutide at certain concentration ratios may result in enhanced IL-6 reduction compared to the individual compounds in this TNFa-induced inflammation model. These findings may indicate potential benefits of combining these compounds for reducing inflammatory responses in certain conditions.

[0289] Referring to FIG. 8, the effects of combining AR1001 and Exenatide on IL-6 level reduction in primary astrocytes are illustrated in two bar graphs. Graph A shows the IL- 6 level reduction in response to Ap+IFNy treatment, while Graph B shows the IL-6 level reduction in response to STZ treatment. In some aspects, the combination of AR1001 and Exenatide may produce a greater reduction in IL-6 levels compared to either compound alone in both experimental conditions.

[0290] In Graph A, the treatments may include Ap+IFNy alone, ARI 001 at 1 pM, Exenatide at 0.1 and 1 pM, and combinations of ARI 001 and Exenatide. The combination of AR1001 and Exenatide may result in a significantly higher reduction of IL-6 levels compared to the individual treatments. In some cases, the combination of 1 pM AR1001 and 0.1 pM Exenatide may produce the greatest reduction in IL-6 levels in the Ap+ZFNy-induced inflammation model.

[0291 ] Graph B follows a similar pattern but with STZ as the initial treatment. The combination of AR 1001 and Exenatide may again demonstrate a synergistic effect in reducing IL-6 levels compared to either compound alone. In some aspects, the combination of 1 pM ARI 001 and 0.1 pM Exenatide may show the most pronounced reduction in IL-6 levels in the STZ-induced inflammation model.

[0292] The error bars in both graphs may indicate variability in the measurements across different samples or replicates. The asterisks above certain bars may denote statistical significance, suggesting that the observed differences in IL-6 reduction between treatment conditions may be statistically meaningful.

[0293] In some cases, the synergistic effects observed in both inflammation models may suggest that the combination of AR 1001 and Exenatide could potentially offer enhanced anti-inflammatory benefits in primary astrocytes compared to monotherapy with either compound. This combination approach may have implications for addressing neuroinflammation associated with various neurodegenerative conditions.

[0294] Referring to Table 20 of the Examples below, the effects of combining AR1001 and Exenatide on IL-6 levels in an Ap+IFNy-induced inflammation model are presented. The data shows the IL-6 fold change and reduction percentages for various concentrations of AR 1001 and Exenatide, both individually and in combination.

[0295] In some aspects, ARI 001 alone at 1 pM may slightly increase IL-6 levels, as indicated by a negative reduction percentage of -2.75%. Exenatide alone may exhibit varying effects depending on the concentration. At 0.1 pM, Exenatide may increase IL-6 levels by 5.41%, while at 1 pM, it may reduce IL-6 levels by 8.52%.

[0296] The combination of AR 1001 and Exenatide may demonstrate synergistic effects in reducing IL-6 levels. For instance, when 1 pM AR1001 is combined with 0.1 pM Exenatide (10: 1 ratio), the IL-6 reduction may reach 22.19%. This combination may exhibit a synergistic effect value of 4.04, indicating a significantly enhanced effect compared to the individual compounds.

[0297] In some cases, the combination of 1 pM AR1001 and 1 pM Exenatide (1 : 1 ratio) may result in an IL-6 reduction of 11.45%, with a synergistic effect value of 1.40. This suggests that even at equal concentrations, AR 1001 and Exenatide may work synergistically to reduce IL-6 levels in this inflammation model.

[0298] The synergistic effect values greater than 1 observed for both combinations indicate that the combined effect of AR 1001 and Exenatide may be greater than the sum of their individual effects. This synergistic interaction may potentially enhance the antiinflammatory properties of these compounds when used in combination.

[0299] Table 21 of the Examples below presents data on the effects of combining AR1001 and Exenatide on IL-6 levels in an STZ-induced inflammation model using primary astrocytes. The table shows the results of various treatments, including STZ alone, AR1001 alone, Exenatide alone, and combinations of AR1001 and Exenatide at different concentrations.

[0300] In some aspects, ARI 001 at a concentration of 1 pM may reduce IL-6 levels by 5.29% compared to the STZ-only control. Exenatide alone at concentrations of 0.1 pM and 1 pM may increase IL-6 levels slightly, by 2.00% and 2.08% respectively.

[0301] The combination of AR 1001 and Exenatide may produce synergistic effects in reducing IL-6 levels. For example, when 1 pM AR1001 is combined with 0.1 pM Exenatide (a 10: 1 ratio), the IL-6 reduction may reach 24.06%. This combination may exhibit a synergistic effect value of 9.46, indicating a substantial enhancement in IL-6 reduction compared to the individual compounds.

[0302] In some cases, a different ratio of AR 1001 to Exenatide may also demonstrate synergistic effects. The combination of 1 pM ARI 001 and 1 pM Exenatide (a 1: 1 ratio) may reduce IL-6 levels by 6.57%, with a synergistic effect value of 1.39.

[0303] These results suggest that the combination of AR1001 and Exenatide may provide enhanced anti-inflammatory effects in STZ-induced inflammation compared to either compound alone. The synergistic effect may vary depending on the concentration ratio of the two compounds, with the 10: 1 ratio showing a particularly strong synergistic interaction in this model.

[0304] Referring to FIG. 9, a bar graph is shown illustrating the effects of AR1001 and semaglutide on JC-1 ratio in an Ap-induced model of mitochondrial dysfunction. The JC- 1 ratio, represented as a percentage on the y-axis, may be indicative of mitochondrial membrane potential. In some aspects, the x-axis displays various treatment conditions, including Ap alone and different combinations of ARI 001 and semaglutide concentrations.

[0305] The graph may demonstrate that combinations of AR 1001 and semaglutide result in higher JC-1 ratios compared to Ap treatment alone or individual compound treatments. In some cases, the combination of AR1001 at 0.1 pM and semaglutide at 0.01 pM may produce a particularly notable increase in JC-1 ratio, suggesting a potential synergistic effect on mitochondrial function. [0306] ARI 001 concentrations in the combinations may range from 0.1 pM to 5 pM, while semaglutide concentrations may range from 0.01 pM to 1 pM. The data suggests that certain combinations of AR 1001 and semaglutide may have synergistic effects in improving mitochondrial function in the presence of Ap. For example, the combination of 0.1 pM ARI 001 and 0.01 pM semaglutide may show a more substantial increase in JC-1 ratio compared to what might be expected from the sum of their individual effects.

[0307] In some aspects, the graph may include error bars for each condition, indicating variability in the measurements across different samples or replicates. The presence of these error bars may provide information about the reliability and consistency of the observed effects.

[0308] The synergistic effects observed in the combination treatments may suggest that AR 1001 and semaglutide could work through complementary mechanisms to protect mitochondrial function in the presence of Ap. This combination approach may offer potential advantages in addressing mitochondrial dysfunction associated with certain neurodegenerative conditions.

[0309] Referring to Table 22, the effects of combining AR1001 and semaglutide on JC-1 ratio in an Ap-induced model are presented. The table shows data for various concentrations of AR1001 and semaglutide, both individually and in combination. In some aspects, AR1001 alone may have limited effects on increasing the JC-1 ratio, with changes ranging from 0 to +7 at concentrations of 0.1 to 5 pM. Similarly, semaglutide alone may show modest increases in JC-1 ratio, with changes of +1 to +14 at concentrations of 0.01 to 1 pM.

[0310] In some cases, the combination of AR 1001 and semaglutide may result in synergistic effects on increasing the JC-1 ratio. For example, the combination of 0.1 pM AR1001 and 0.01 pM semaglutide may lead to a 28% increase in JC-1 ratio, with a synergistic effect value of 23.74. This synergistic effect value, calculated as (AB/A+B), may indicate a strong positive interaction between AR1001 and semaglutide in improving mitochondrial function.

[0311] The data suggests that various concentration ratios of AR1001 to semaglutide may produce synergistic effects. In some aspects, ratios ranging from 1: 1 to 100: 1 may be effective. For instance, a 10: 1 ratio (0.1 pM AR1001 to 0.01 pM semaglutide) may yield a synergistic effect value of 23.74, while a 1 : 1 ratio (0.1 pM ARlOOl to O. l pM semaglutide) may result in a synergistic effect value of 2.40.

[0312] Higher concentrations of AR 1001 combined with semaglutide may also demonstrate synergistic effects. For example, 5 pM ARI 001 combined with 1 pM semaglutide may increase the JC-1 ratio by 21%, with a synergistic effect value of 1.27. These results suggest that the combination of AR1001 and semaglutide may have potential benefits in mitigating Ap-induced mitochondrial dysfunction across a range of concentration ratios.

[0313] Referring to FIG. 10, a bar graph is shown depicting the effects of AR1001 and Exenatide on JC-1 ratio in primary neurons exposed to Ap. The JC-1 ratio may be used as an indicator of mitochondrial membrane potential, with higher ratios suggesting improved mitochondrial function. In some aspects, the graph displays the JC-1 ratio as a percentage on the y-axis, ranging from 0 to 100%. The x-axis may show different treatment conditions, including Ap alone, ARI 001 at concentrations of 0.1 and 0.5 pM, and combinations of AR1001 and Exenatide at various concentrations.

[0314] In some cases, the treatments with combined AR 1001 and Exenatide may show higher JC-1 ratios compared to Ap alone or individual treatments. This suggests that the combination of AR 1001 and Exenatide may have a synergistic effect in maintaining mitochondrial membrane potential in primary neurons exposed to Ap. The graph may include error bars for each condition, indicating the variability of the measurements across different samples or replicates.

[0315] The combination of 0.1 pM AR1001 and 0.001 pM Exenatide may result in a higher JC-1 ratio compared to either compound alone. Similarly, the combination of 0.5 pM AR1001 and 0.001 pM Exenatide may show an even greater increase in JC-1 ratio. These results suggest that AR 1001 and Exenatide may work synergistically to protect mitochondrial function in the presence of Ap-induced stress.

[0316] In some aspects, the synergistic effect observed with the combination of AR 1001 and Exenatide may be due to complementary mechanisms of action. AR 1001, as a PDE5 inhibitor, may increase intracellular cGMP levels, while Exenatide, as a GLP-1 receptor agonist, may activate neuroprotective signaling pathways. The combination of these effects may lead to enhanced mitochondrial protection in primary neurons exposed to Ap.

[0317] The data presented in FIG. 10 may provide evidence for the potential therapeutic benefits of combining AR1001 and Exenatide in treating conditions associated with mitochondrial dysfunction, such as neurodegenerative disorders. However, it is important to note that the optimal concentrations and ratios of AR 1001 and Exenatide for achieving maximal synergistic effects may vary depending on the specific cellular context and experimental conditions. [0318] Referring to Table 23, the effects of combining AR1001 and Exenatide on JC-1 ratio in an Ap-induced model of mitochondrial dysfunction in primary neurons are presented. The table shows data for various concentrations of AR1001 and Exenatide, both individually and in combination, in the presence of 10 pM Ap. In some aspects, the JC-1 ratio may be used as an indicator of mitochondrial membrane potential, with higher ratios suggesting improved mitochondrial function.

[0319] The data indicates that ARI 001 alone at 0.1 pM may slightly decrease the JC-1 ratio by 1%, while at 0.5 pM it may increase the ratio by 6% compared to the Ap-only control. Exenatide alone at 0.001 pM may increase the JC-1 ratio by 7%.

[0320] When AR1001 and Exenatide are combined, synergistic effects on JC-1 ratio may be observed. For instance, the combination of 0.1 pM AR1001 and 0.001 pM Exenatide may result in a 6% increase in JC-1 ratio, while 0.5 pM AR1001 and 0.001 pM Exenatide may lead to a 15% increase. These combination effects may be greater than the sum of the individual compound effects, as indicated by the synergistic effect values of 1.15 and 1.17, respectively.

[0321] The concentration ratios of AR1001 to Exenatide tested in this study were 100: 1 and 500: 1. In some cases, these ratios may provide optimal synergistic effects for improving mitochondrial function in the presence of Ap. The data suggests that combining ARI 001 and Exenatide may offer potential benefits in mitigating Ap-induced mitochondrial dysfunction in primary neurons.

[0322] Referring to FIG. 11, a bar graph is shown illustrating the effects of AR1001 and Semaglutide combination therapy on Ap reduction in SH-SY5Y cells. The graph displays Ap reduction percentages for various concentrations of ARI 001 and Semaglutide, both individually and in combination. In some aspects, the combination of AR1001 and Semaglutide may result in greater Ap reduction compared to either compound alone. The highest Ap reduction may be observed for certain concentration ratios of ARI 001 and Semaglutide.

[0323] In some cases, AR1001 and Semaglutide may exhibit synergistic effects on Ap reduction at specific concentration ratios. For example, the combination may show synergistic effects at AR1001 to Semaglutide ratios of 10:3, 10:7, 10: 14, 30:3, 30:7, and 30: 14. These ratios may correspond to oral doses of 3, 7, and 14 mg/day for Semaglutide, and 10 and 30 mg/day for AR 1001.

[0324] The graph may include error bars for each treatment condition, indicating variability in the measurements. Statistical significance may be denoted by symbols explained in a legend accompanying the graph. The x-axis of the graph may display different concentrations of AR1001 and Semaglutide, while the y-axis may represent Ap reduction as a percentage.

[0325] In some aspects, the combination of AR1001 and Semaglutide may provide enhanced Ap reduction compared to monotherapy with either compound. This enhanced effect may be particularly pronounced at certain concentration ratios, which may correspond to specific oral dosing regimens. The data presented in FIG. 11 may suggest potential benefits of combining ARI 001 and Semaglutide for reducing Ap levels in cellular models relevant to neurodegenerative conditions.

[0326] Referring to Table 24, the combination of mirodenafil AR 1001 and semaglutide may exhibit synergistic effects in reducing intracellular Ap levels. In some aspects, the combination of mirodenafil AR1001 and semaglutide at certain concentrations may result in a synergistic effect value of 2.87. This synergistic effect value may indicate a significant enhancement in the reduction of intracellular Ap levels compared to the individual effects of mirodenafil AR 1001 and semaglutide alone.

Table 24

A: Ap+ARlOOl treated group; B: Ap+ Semaglutide treated group; AB: combined treatment of AB+AR1001+ Semaglutide Synergistic Effect Evaluation (AB / A + B)

> 1 Synergistic Effect; = 1 Additive Effect; < 1 Antagonistic Effect

[0327] In some cases, the combination of mirodenafil AR 1001 and semaglutide may produce a synergistic effect value of 2.57 in reducing intracellular Ap levels. This synergistic effect value may suggest a substantial improvement in Ap reduction when mirodenafil AR1001 and semaglutide are used in combination at specific concentrations.

[0328] The data in Table 24 may also demonstrate a synergistic effect value of 2.14 for certain combinations of mirodenafil AR 1001 and semaglutide. This synergistic effect value may indicate that the combination therapy may be more effective in reducing intracellular Ap levels than would be expected from the sum of their individual effects.

[0329] In some aspects, the combination of mirodenafil AR 1001 and semaglutide may result in a synergistic effect value of 1.78. This synergistic effect value may suggest that the combination therapy may provide enhanced efficacy in reducing intracellular Ap levels compared to monotherapy with either compound.

[0330] The use of mirodenafil AR1001 as the specific PDE5 inhibitor in combination with semaglutide may contribute to these observed synergistic effects. Mirodenafil AR 1001 may interact with semaglutide in a manner that enhances their combined ability to reduce intracellular Ap levels. The synergistic effects observed at various concentration combinations may suggest that the interaction between mirodenafil AR 1001 and semaglutide may be concentration-dependent and may be optimized for maximal Ap reduction.

[0331] In some aspects, the combination of AR 1001 and semaglutide may exhibit a synergistic effect value of 1.24 in reducing intracellular Ap levels. This synergistic effect may be observed when specific concentrations of AR1001 and semaglutide are combined. The synergistic effect value greater than 1 suggests that the combination of AR 1001 and semaglutide may produce a greater reduction in intracellular Ap levels than would be expected from the sum of their individual effects.

[0332] In other cases, the combination of AR1001 and semaglutide may demonstrate a synergistic effect value of 1.44 in reducing intracellular Ap levels. This higher synergistic effect value may indicate an even more pronounced cooperative action between AR1001 and semaglutide in reducing Ap accumulation within cells. The specific concentrations of AR1001 and semaglutide that result in this synergistic effect may vary, and optimal ratios may be determined through experimentation.

[0333] These synergistic effects may suggest that the combination of AR1001 and semaglutide could potentially provide enhanced therapeutic benefits in conditions associated with elevated intracellular Ap levels. The observed synergy may allow for the use of lower doses of each compound while still achieving significant reductions in Ap levels, which may potentially lead to improved efficacy and reduced side effects.

[0334] Referring to FIG. 12, a bar graph illustrates the effects of AR1001 and Semaglutide combination therapy on ILip reduction in BV-2 cells. The graph displays ILip reduction percentages for various concentrations of AR1001 and Semaglutide, both individually and in combination. In some aspects, the combination of AR1001 and Semaglutide may result in greater ILip reduction compared to either compound alone.

[0335] The x-axis of the graph shows different concentrations of ARI 001 (2 pM and 6 pM) and Semaglutide (0.6 pM, 1.4 pM, and 2.8 pM), while the y-axis represents the ILip reduction percentage. Multiple bars are displayed for each treatment condition, with error bars indicating variability in the measurements.

[0336] In some cases, ARI 001 alone at 2 pM and 6 pM concentrations may produce modest reductions in ILip levels. Semaglutide alone at 0.6 pM, 1.4 pM, and 2.8 pM concentrations may also exhibit some ILip reduction effects. However, when AR1001 and Semaglutide are combined, the ILip reduction may be significantly enhanced.

[0337] The graph suggests that certain concentration ratios of AR1001 to Semaglutide may be particularly effective in reducing ILip levels. For example, the combination of 6 pM AR1001 with 0.6 pM Semaglutide may result in a substantial reduction in ILip levels, potentially indicating a synergistic effect. Similarly, the combination of 6 pM AR1001 with 1.4 pM Semaglutide may also demonstrate enhanced ILip reduction compared to either compound alone.

[0338] In some aspects, the synergistic effects of ARI 001 and Semaglutide on ILip reduction may be observed across multiple concentration ratios. The graph indicates that combinations of AR1001 and Semaglutide at ratios of 10:3, 10:7, 30:3, 30:7, and 30: 14 may all exhibit enhanced ILip reduction compared to the individual compounds.

[0339] The data presented in FIG. 12 may suggest that combining AR 1001 and Semaglutide could potentially offer improved anti-inflammatory effects in BV-2 cells, as indicated by the reduction in ILip levels. This combination approach may provide a basis for developing more effective strategies to address inflammatory processes in certain cellular contexts.

[0340] Referring to Table 25, the effects of combining AR 1001 and semaglutide on ILip levels in an LPS-induced inflammation model are presented. The table shows data for various concentrations of AR1001 and semaglutide, both individually and in combination. In some aspects, AR1001 alone may reduce ILip levels in a concentration-dependent manner. For example, 2 pM AR1001 may result in a 0.41% reduction, while 6 pM AR1001 may lead to a 17.43% reduction in ILip levels.

Table 25

treatment of LPS +AR1001+Semaglutide Synergistic Effect Evaluation (AB / A + B) > 1 Synergistic Effect; = 1 Additive Effect; < 1 Antagonistic Effect

[0341 ] Semaglutide alone may also exhibit concentration-dependent effects on ILip reduction. In some cases, 0.6 pM semaglutide may reduce ILip levels by 9.85%, while 1.4 pM and 2.8 pM semaglutide may result in 32.28% and 56.66% reductions, respectively.

[0342] When AR1001 and semaglutide are combined, synergistic effects on ILip reduction may be observed. For instance, the combination of 2 pM AR1001 with 0.6 pM semaglutide may result in a 14.52% reduction in ILip levels, with a synergistic effect value of 1.42. This synergistic effect value, being greater than 1, suggests that the combination may be more effective than the sum of the individual effects of AR1001 and semaglutide alone.

[0343] In some aspects, higher concentrations of AR 1001 combined with semaglutide may lead to more pronounced synergistic effects. For example, 6 pM AR1001 combined with 0.6 pM semaglutide may result in a 56.12% reduction in ILip levels, with a synergistic effect value of 2.06. This combination may demonstrate the highest synergistic effect among the tested concentrations.

[0344] The data in Table 25 may indicate that various concentration ratios of ARI 001 and semaglutide can produce synergistic effects in reducing ILip levels. In some cases, the synergistic effect values range from 1.42 to 2.06, depending on the specific concentration combinations. These results suggest that combining AR1001 and semaglutide may provide enhanced anti-inflammatory effects compared to either compound used alone in this LPS -induced inflammation model.

[0345] Referring to FIG. 13, a bar graph is shown illustrating the effects of AR1001 and Semaglutide combination therapy on TNFa reduction in BV-2 cells. The graph displays TNFa reduction percentages for various concentrations of ARI 001 and Semaglutide, both individually and in combination. In some aspects, the combination of AR1001 and Semaglutide may result in greater TNFa reduction compared to either compound alone.

[0346] The x-axis of the graph shows different treatment conditions, including varying concentrations of AR1001 (which may range from 2 pM to 6 pM) and Semaglutide (which may range from 0.6 pM to 2.8 pM). The y-axis represents the TNFa reduction as a percentage, which may range from 0 to 100%. Multiple bars are displayed for each treatment condition, with error bars indicating variability in the measurements.

[0347] In some cases, the highest TNFa reduction may be observed for combinations of AR1001 and Semaglutide at certain concentration ratios. For example, the combination of 6 pM AR1001 and 2.8 pM Semaglutide may show a particularly strong reduction in TNFa levels. This combination may demonstrate a synergistic effect, where the combined treatment produces a greater reduction in TNFa than would be expected from the sum of their individual effects.

[0348] The graph may include a legend explaining the symbols used to denote statistical significance and different treatment conditions. This information can be useful for interpreting the relative effectiveness of various treatment combinations.

[0349] In some aspects, AR1001 and Semaglutide may have a synergistic effect on down-regulation of TNFa expression at specific ratios. For instance, the ratios of AR1001 to Semaglutide that may show synergistic effects could include 10:3, 10:7, 10: 14, 30:3, 30:7, and 30: 14. These ratios may correspond to the concentrations used in the study and may provide insights into the optimal combinations for TNFa reduction.

[0350] The synergistic effects observed in this study may suggest that combining AR1001 and Semaglutide could potentially enhance anti-inflammatory effects in BV-2 cells. This combination therapy approach may offer advantages over single-agent treatments in reducing TNFa levels, which may be relevant in the context of neuroinflammatory conditions.

[0351 ] Referring to Table 26, the data illustrates the effects of combining mirodenafil ARI 001 and semaglutide on TNFa levels in an LPS-induced inflammation model. The table presents results for various concentrations of mirodenafil AR1001 and semaglutide, both individually and in combination. In some aspects, mirodenafil AR1001 alone may reduce TNFa levels to varying degrees depending on the concentration used. For example, at 2 pM, AR1001 may reduce TNFa levels by 6.94%, while at 6 pM, the reduction may increase to 8.92%. Similarly, semaglutide alone may exhibit concentration-dependent effects on TNFa reduction, with 0.6 pM, 1.4 pM, and 2.8 pM concentrations resulting in 8.15%, 23.57%, and 39.78% reductions, respectively. Table 26

A: LPS+AR1001 treated group; B: LPS+Semaglutide treated group; AB: combined treatment of LPS+ARlOOl+Semaglutide Synergistic Effect Evaluation (AB / A + B)

> 1 Synergistic Effect; =1 Additive Effect; < 1 Antagonistic Effect

[0352] The combination of mirodenafil AR 1001 and semaglutide may produce synergistic effects in reducing TNFa levels. In some cases, when 2 pM mirodenafd AR1001 is combined with 0.6 pM semaglutide, a synergistic effect value 1.07 may be observed, indicating a greater reduction in TNFa levels than what would be expected from the sum of their individual effects. The synergistic effect may become more pronounced at certain concentration ratios. For instance, the combination of 6 pM mirodenafil ARlOOl with 0.6 pM semaglutide may result in a synergistic effect value 1.16, suggesting an enhanced ability to reduce TNFa levels.

[0353] In some aspects, the synergistic effect may be further increased by adjusting the concentration ratios of mirodenafil ARlOOl and semaglutide. The data shows that combining 6 pM mirodenafil ARlOOl with 1.4 pM semaglutide may lead to a synergistic effect value 1.16, potentially indicating an optimal ratio for maximizing the reduction of TNFa levels in this inflammation model. These synergistic effects may suggest that the combination of mirodenafil ARlOOl and semaglutide could provide enhanced antiinflammatory benefits compared to either compound used alone.

[0354] Referring to FIG. 14, a combination bar graph and data table are shown illustrating the effects of ARlOOl and Sitagliptin on Ap reduction. The bar graph displays Ap reduction percentages on the y-axis and different concentrations of ARlOOl and Sitagliptin on the x-axis. In some aspects, various colored bars may represent different treatment conditions. The highest reduction may be observed for combinations of ARlOOl and Sitagliptin.

[0355] In some cases, the data table below the graph may provide detailed information on ARlOOl and Sitagliptin concentrations, Ap concentrations, reduction rates, and AB/(A+B) ratios. The table may include calculations for synergistic effects, with values greater than 1 potentially indicating synergistic interactions between ARlOOl and Sitagliptin in reducing Ap levels.

[0356] The combination of ARlOOl and Sitagliptin may demonstrate enhanced Ap reduction compared to either compound alone. For example, in some aspects, ARlOOl at 0.2 pM combined with Sitagliptin at 1 pM may result in a 5.83% reduction in Ap levels, while ARlOOl or Sitagliptin alone at these concentrations may show lower reduction rates of 2.36% and 0.75%, respectively.

[0357] In some cases, the most effective concentration ratios for ARlOOl to Sitagliptin may range from 1 :3.3 to 1 : 10. For instance, the combination of 0.2 pM ARlOOl and 2 pM Sitagliptin (1 : 10 ratio) may exhibit a 14.52% reduction in Ap levels, with an AB/(A+B) ratio of 1.235, suggesting a potential synergistic effect.

[0358] The statistical significance of the results may be indicated by symbols in the legend above the graph. These symbols may denote different levels of significance when comparing the combination treatments to individual compound treatments or controls.

[0359] In some aspects, the synergistic effects of ARlOOl and Sitagliptin combinations may be concentration-dependent. For example, higher concentrations of both compounds may not necessarily lead to greater synergistic effects. The data may suggest that optimal synergy may be achieved at specific concentration ratios, which may be important for potential therapeutic applications.

[0360] The AB/(A+B) ratios provided in the data table may offer a quantitative measure of the synergistic effects. Values greater than 1 may indicate synergy, with higher values potentially suggesting stronger synergistic interactions. In some cases, these ratios may help identify the most promising combination ratios for further investigation or development.

[0361 ] Referring to FIG. 15, a combination bar graph and data table are shown illustrating the effects of ARI 001 and Vildagliptin on Ap reduction. The bar graph displays Ap reduction percentages on the y-axis and different concentrations of AR1001 and Vildagliptin on the x-axis. In some aspects, various colored bars may represent different treatment conditions. The highest reduction may be observed for combinations of AR1001 and Vildagliptin at 0.6 pM and 2.0 pM respectively.

[0362] A legend above the graph may explain the symbols used to denote statistical significance and treatment conditions. In some cases, the data table below the graph provides detailed information on ARI 001 and Vildagliptin concentrations, Ap concentrations, reduction rates, and AB/(A+B) ratios. The table may include calculations for synergistic effects, with values greater than 1 indicating synergistic interactions between AR1001 and Vildagliptin in reducing Ap levels.

[0363] In some aspects, the combination of AR1001 and Vildagliptin may demonstrate synergistic effects in reducing Ap levels compared to either compound alone. The most effective concentration ratios may be observed when ARI 001 is at 0.6 pM and Vildagliptin is at 2.0 pM. This combination may result in a higher Ap reduction percentage compared to other concentration combinations or individual treatments.

[0364] The statistical significance of the results may be indicated by symbols in the bar graph. In some cases, these symbols may denote different levels of significance, allowing for a quick visual assessment of which combinations produce statistically significant reductions in Ap levels compared to controls or individual treatments.

[0365] The AB/(A+B) ratios provided in the data table may offer a quantitative measure of the synergistic effects. Values greater than 1 may suggest that the combination of AR1001 and Vildagliptin produces a greater effect than would be expected from the sum of their individual effects. In some aspects, this synergistic effect may be particularly pronounced at certain concentration ratios, which may be identified by examining the AB/(A+B) values across different combinations.

[0366] Referring to FIG. 16, a combination bar graph and data table are shown illustrating the effects of ARI 001 and Linagliptin on Ap reduction. The bar graph displays Ap reduction percentages on the y-axis, ranging from 0 to 100%, and different concentrations of AR1001 and Linagliptin on the x-axis. In some aspects, various colored bars may represent different treatment conditions. The highest reduction may be observed for the combination of 0.6 pM ARI 001 and 0.1 pM Linagliptin.

[0367] In some cases, a legend above the graph may explain the symbols used to denote statistical significance. The data table below the graph may provide detailed information on AR1001 and Linagliptin concentrations, Ap concentrations, reduction rates, and AB/(A+B) ratios. The table may include calculations for synergistic effects, with values greater than 1 potentially indicating synergistic interactions between AR 1001 and Linagliptin in reducing Ap levels.

[0368] The combination of AR 1001 and Linagliptin may demonstrate synergistic effects in reducing Ap levels compared to either compound alone. For example, ARI 001 at 0.2 pM alone may result in a 2.36% reduction in Ap levels, while Linagliptin at 0.1 pM alone may result in a 6.29% reduction. However, when combined at these concentrations, the reduction may increase to 15.73%, with an AB/(A+B) ratio of 1.819, suggesting a synergistic effect.

[0369] In some aspects, the most effective concentration ratio of AR1001 to Linagliptin may be 6: 1, as demonstrated by the combination of 0.6 pM AR1001 and 0.1 pM Linagliptin. This combination may result in a 46.50% reduction in Ap levels, with an AB/(A+B) ratio of 1.231, indicating a strong synergistic effect.

[0370] The statistical significance of the results may be indicated by symbols in the bar graph. In some cases, asterisks or other markers may denote levels of significance, such as p < 0.05, p < 0.01, or p < 0.001, when comparing the combination treatments to individual compound treatments or controls.

[0371] A note at the bottom of the figure may explain how to interpret the AB/(A+B) ratios. Values less than 1 may indicate an antagonistic effect, values equal to 1 may suggest an additive effect, and values greater than 1 may indicate a synergistic effect between ARI 001 and Linagliptin in reducing Ap levels.

[0372] The data presented in FIG. 16 may demonstrate the potential of combining ARI 001 and Linagliptin at specific concentration ratios to achieve enhanced Ap reduction compared to either compound alone. This synergistic effect may have implications for the development of more effective treatments for conditions associated with Ap accumulation.

[0373] Referring to FIG. 17, a bar graph is shown illustrating the effects of AR1001 and Sitagliptin on ILip levels in an LPS-induced inflammation model. The graph displays ILip mRNA fold change on the y-axis, ranging from 0 to 18, while the x-axis shows different treatment conditions. In some aspects, the treatment conditions may include LPS alone, varying concentrations of ARI 001 (2 pM and 6 pM), and combinations of ARI 001 with Sitagliptin (10 pM and 20 pM).

[0374] The graph demonstrates that combinations of AR1001 and Sitagliptin may result in lower ILip mRNA fold changes compared to LPS alone or individual treatments. In some cases, the lowest ILip levels may be observed for the combination of 6 pM ARI 001 and 20 pM Sitagliptin. This combination may exhibit a synergistic effect in reducing ILip levels, as the reduction appears to be greater than the sum of the reductions achieved by each compound individually.

[0375] The data suggests that AR1001 alone may have a dose-dependent effect on ILip reduction, with 6 pM showing a greater reduction than 2 pM. Similarly, Sitagliptin alone may also demonstrate a dose-dependent effect, with 20 pM potentially being more effective than 10 pM in reducing ILip levels.

[0376] When AR1001 and Sitagliptin are combined, the reduction in ILip levels may be more pronounced. For instance, the combination of 2 pM AR1001 with 10 pM Sitagliptin may show a greater reduction in ILip levels compared to either compound alone at these concentrations. The synergistic effect appears to be further enhanced when higher concentrations of both compounds are used in combination.

[0377] Error bars included for each condition indicate the variability of the measurements, which may be considered when interpreting the results. The statistical significance of the differences between treatment groups may be denoted by symbols explained in the legend, allowing for a more detailed analysis of the synergistic effects observed.

[0378] In some aspects, these results may suggest that the combination of AR 1001 and Sitagliptin could potentially offer improved anti-inflammatory effects in conditions where ILip plays a role, such as in certain neurodegenerative disorders or inflammatory diseases. The synergistic interaction between AR1001 and Sitagliptin may allow for lower doses of each compound to be used while still achieving significant reductions in inflammatory markers. [0379] Referring to FIG. 18, a bar graph illustrates the effects of AR 1001 and Sitagliptin on ILip mRNA reduction in an LPS-induced inflammation model. The graph displays ILip mRNA reduction percentages for various concentrations of AR1001 and Sitagliptin, both individually and in combination. In some aspects, the y-axis represents ILip mRNA reduction as a percentage, ranging from 0 to 100%. The x-axis may show different concentrations of ARI 001 (2 pM and 6 pM) and Sitagliptin (10 pM and 20 pM).

[0380] The graph demonstrates that combinations of AR1001 and Sitagliptin may result in higher ILip mRNA reduction percentages compared to either compound alone. In some cases, the highest reduction may be observed for the combination of 6 pM ARI 001 and 20 pM Sitagliptin. The data suggests potential synergistic effects between AR1001 and Sitagliptin in reducing ILip mRNA levels.

[0381] Multiple bars may be shown for each treatment condition, with error bars indicating variability in the measurements. A legend in the upper right corner of the graph may explain the symbols used to denote statistical significance for different comparisons. The statistical significance indicators allow for the evaluation of the reliability and importance of the observed differences between treatment conditions.

[0382] In some aspects, the graph may reveal a dose-dependent effect for both AR1001 and Sitagliptin when used individually. The combination treatments may show enhanced ILip mRNA reduction compared to the sum of the individual effects, suggesting a potential synergistic interaction between AR 1001 and Sitagliptin in modulating inflammatory responses.

[0383] The data presented in FIG. 18 may provide insights into the potential antiinflammatory effects of combining AR1001 and Sitagliptin. This combination approach may offer advantages in reducing ILip mRNA levels compared to monotherapy with either compound. The observed synergistic effects may have implications for developing more effective strategies to manage inflammatory conditions.

[0384] Referring to FIG. 19, a bar graph is shown illustrating the effects of AR1001 and Sitagliptin on TNFa mRNA levels in an LPS-induced inflammation model. The y-axis of the graph represents TNFa mRNA fold change, while the x-axis displays different treatment conditions. In some aspects, the graph may demonstrate that combinations of AR1001 and Sitagliptin result in lower TNFa mRNA fold changes compared to LPS alone or individual treatments.

[0385] The graph may show results for various concentrations of ARI 001 (2 pM and 6 pM) and Sitagliptin (10 pM and 20 pM), both individually and in combination. In some cases, the lowest TNFa levels may be observed for the combination of 6 pM AR1001 and 20 pM Sitagliptin. This combination may exhibit a synergistic effect in reducing TNFa mRNA levels beyond what might be expected from the individual effects of AR1001 and Sitagliptin alone.

[0386] Error bars included for each condition may indicate the variability of the measurements. A legend in the upper right corner of the graph may explain the statistical significance symbols used, allowing for comparison between different treatment groups.

[0387] In some aspects, the graph may demonstrate a dose-dependent effect of AR1001 on TNFa mRNA reduction, with 6 pM AR1001 showing a greater effect than 2 pM ARI 001. Similarly, Sitagliptin at 20 pM may exhibit a stronger TNFa-reducing effect compared to 10 pM Sitagliptin.

[0388] The combination treatments may show enhanced TNFa mRNA reduction compared to either compound alone. For example, the combination of 2 pM AR1001 with 10 pM Sitagliptin may result in a greater reduction of TNFa mRNA levels than either 2 pM ARI 001 or 10 pM Sitagliptin individually. This synergistic effect may be even more pronounced for the combination of 6 pM ARI 001 and 20 pM Sitagliptin.

[0389] In some cases, the data presented in FIG. 19 may suggest that combining AR1001 and Sitagliptin could potentially provide enhanced anti-inflammatory effects in conditions where TNFa plays a role. The synergistic reduction in TNFa mRNA levels observed with certain combinations of AR1001 and Sitagliptin may indicate a possible therapeutic strategy for managing inflammatory conditions.

[0390] Referring to FIG. 20, a bar graph illustrates the effects of AR 1001 and Sitagliptin on TNFa mRNA reduction in an LPS-induced inflammation model. The graph displays TNFa mRNA reduction percentages for different concentrations of ARI 001 (2 pM and 6 pM) and Sitagliptin (10 pM and 20 pM), both individually and in combination. In some aspects, the combinations of ARI 001 and Sitagliptin may result in higher TNFa reduction percentages compared to either compound alone. The highest reduction may be observed for the combination of 6 pM ARI 001 and 20 pM Sitagliptin.

[0391] Error bars in the graph indicate variability in the measurements across different samples or replicates. A legend in the upper right comer explains the symbols used to denote statistical significance for different comparisons. In some cases, the statistical significance symbols may indicate that the observed differences between treatment conditions are unlikely to have occurred by chance.

[0392] The synergistic effects of combining ARI 001 and Sitagliptin on TNFa mRNA reduction may be particularly evident when comparing the combination treatments to the individual compound treatments. For example, while ARI 001 at 6 pM or Sitagliptin at 20 pM alone may produce moderate reductions in TNFa mRNA levels, their combination may lead to a substantially greater reduction, suggesting a potential synergistic interaction between the two compounds in modulating the inflammatory response.

[0393] In some aspects, the concentration-dependent effects of AR1001 and Sitagliptin on TNFa mRNA reduction may be observed. The graph may show that increasing concentrations of either compound alone may lead to greater reductions in TNFa mRNA levels. However, the most pronounced effects may be seen when higher concentrations of both compounds are combined.

[0394] The data presented in FIG. 20 may provide insights into the potential antiinflammatory mechanisms of AR1001 and Sitagliptin when used in combination. The observed synergistic effects on TNFa mRNA reduction may suggest that these compounds may work through complementary pathways to modulate the inflammatory response in this model system.

[0395] In some aspects, combinations of mirodenafil AR 1001 and sitagliptin may exhibit synergistic effects in reducing ILip levels in an LPS-induced inflammation model. Referring to Table 27, various concentrations of mirodenafil AR1001 and sitagliptin were evaluated individually and in combination for their effects on ILip production.

Table 27

[0396] The data indicates that mirodenafil ARI 001 alone at concentrations of 2 pM and 6 pM may reduce ILip levels by 8.07% and 45.15%, respectively. Sitagliptin alone at concentrations of 10 pM and 20 pM may reduce ILip levels by 12.23% and 22.62%, respectively.

[0397] When mirodenafil AR 1001 and sitagliptin are combined, greater reductions in ILip levels may be observed compared to either compound alone. For example, the combination of 2 pM mirodenafil ARI 001 with 10 pM sitagliptin may result in a 23.54% reduction in ILip levels, while the combination of 6 pM mirodenafil AR1001 with 20 pM sitagliptin may lead to a 74.80% reduction.

[0398] The synergistic effects of these combinations may be quantified by the AB/(A+B) ratio, where values greater than 1 suggest synergistic interactions. In some cases, the AB/(A+B) ratios for mirodenafil AR1001 and sitagliptin combinations range from 1.10 to 1.23, indicating potential synergistic effects across various concentration ratios.

[0399] These results suggest that combining mirodenafil AR1001 and sitagliptin may provide enhanced anti-inflammatory effects by reducing ILip levels more effectively than either compound alone in this LPS-induced inflammation model. The synergistic interactions observed may occur at different concentration ratios of mirodenafil AR1001 to sitagliptin, potentially allowing for flexibility in dosing strategies.

[0400] Referring to Table 28, a data table 100 may be used to present results of combination therapy effects on TNFa levels in an LPS-induced inflammation model. The data table 100 may include columns for mirodenafil AR 1001 concentration, sitagliptin concentration, LPS treatment, TNFa fold change, TNFa reduction percentage, and AB/(A+B) ratio. In some aspects, the data table 100 may be organized into sections for mirodenafil AR1001 alone, sitagliptin alone, and combinations of mirodenafil AR1001 and sitagliptin.

Table 28

[0401] The data table may show that mirodenafil ARI 001 alone at 2 pM and 6 pM concentrations may reduce TNFa levels by 5.08% and 22.95% respectively. In some cases, sitagliptin alone at 10 pM and 20 pM concentrations may reduce TNFa levels by 0.49% and 16.63% respectively.

[0402] When mirodenafil AR 1001 and sitagliptin are combined, greater reductions in TNFa levels may be observed. For example, the combination of 2 pM mirodenafil ARI 001 with 10 pM sitagliptin may result in a 6.64% reduction in TNFa levels. In some aspects, the highest reduction of 43.99% may be observed for 6 pM mirodenafil ARI 001 combined with 20 pM sitagliptin.

[0403] The AB/(A+B) ratio may be used to assess potential synergistic effects between mirodenafil AR1001 and sitagliptin. In some cases, these values may range from 1.11 to 1.19 for different concentration combinations. Values greater than 1 may suggest synergistic effects in reducing TNFa levels. For instance, the combination of 2 pM mirodenafil ARI 001 with 10 pM sitagliptin may show an AB/(A+B) value of 1.19, indicating a potential synergistic interaction. [0404] The data presented in the table may demonstrate that combinations of mirodenafil AR1001 and sitagliptin at various concentration ratios may produce greater reductions in TNFa levels compared to either compound alone in this LPS-induced inflammation model.

[0405] Referring to FIG. 21, a bar graph is shown illustrating the effects of AR1001 and Linagliptin on ILip mRNA reduction in an LPS-induced inflammation model. The y-axis of the graph represents ILip mRNA reduction as a percentage, ranging from 0 to 100%. The x-axis displays different concentrations of Linagliptin (0, 1 pM) and ARI 001 (2, 6 pM), both individually and in combination.

[0406] In some aspects, the graph demonstrates that combinations of AR 1001 and Linagliptin may result in higher ILip mRNA reduction percentages compared to either compound alone. The highest reduction may be observed for the combination of 6 pM ARI 001 and 1 pM Linagliptin. This combination may exhibit a synergistic effect, potentially leading to a more pronounced reduction in ILip mRNA levels than what might be expected from the sum of the individual effects of AR 1001 and Linagliptin.

[0407] The graph may include error bars for each condition, which may indicate variability in the measurements across different samples or replicates. These error bars may provide information about the consistency and reliability of the observed effects.

[0408] In some cases, a legend in the upper right corner of the figure may explain the symbols used to denote statistical significance for different comparisons. This information may be useful for interpreting the relative importance of the observed differences between treatment conditions.

[0409] The synergistic effect observed in the combination of AR1001 and Linagliptin may suggest a potential therapeutic advantage in using these compounds together for reducing inflammatory responses. However, it is important to note that the optimal concentration ratio and absolute concentrations of AR1001 and Linagliptin may vary depending on the specific application or physiological context.

[0410] In some aspects, the data presented in FIG. 21 may provide insights into the potential mechanisms by which AR 1001 and Linagliptin interact to modulate inflammatory responses. The enhanced reduction of ILip mRNA levels in the presence of both compounds may indicate complementary or synergistic effects on signaling pathways involved in inflammation regulation.

[0411 ] Referring to FIG. 22, a bar graph is shown illustrating the effects of AR1001 and Linagliptin on TNFa mRNA reduction in an LPS-induced inflammation model. The graph displays TNFa mRNA reduction as a percentage on the y-axis, ranging from 0 to 100%. The x-axis shows different concentrations of Linagliptin (0, 1 pM) and ARI 001 (2, 6 pM), both individually and in combination.

[0412] In some aspects, the graph demonstrates that combinations of AR 1001 and Linagliptin may result in higher TNFa mRNA reduction percentages compared to either compound alone. The highest reduction may be observed for the combination of 6 pM ARI 001 and 1 pM Linagliptin. This combination may exhibit a synergistic effect, potentially leading to a more pronounced reduction in TNFa mRNA levels than what might be expected from the sum of their individual effects.

[0413] The graph includes error bars for each condition, which may indicate variability in the measurements across different samples or replicates. These error bars may provide information about the consistency and reliability of the observed effects.

[0414] In some cases, statistical significance between different treatment conditions may be denoted by symbols, as explained in a legend in the upper left comer of the figure. This statistical analysis may help in determining whether the observed differences between treatment groups are likely to be meaningful rather than due to chance.

[0415] The synergistic effect observed in the combination of AR1001 and Linagliptin may suggest a potential therapeutic advantage in using these compounds together for reducing inflammatory responses. However, it is important to note that the optimal concentration ratio and absolute concentrations of these compounds may vary depending on specific conditions or applications.

[0416] In some aspects, the data presented in this figure may provide insights into the potential anti-inflammatory mechanisms of AR1001 and Linagliptin when used in combination. The reduction in TNFa mRNA levels may indicate a decrease in the expression of this pro-inflammatory cytokine, which could contribute to an overall reduction in inflammatory responses.

[0417] It is worth noting that while this data shows promising results in an LPS- induced inflammation model, further studies may be necessary to fully elucidate the mechanisms underlying the observed synergistic effects and to determine the potential clinical implications of these findings.

[0418] Referring to Table 29, the effects of combining mirodenafil AR 1001 and linagliptin on IL-ip levels in an LPS-induced inflammation model are presented. The data shows that mirodenafil AR1001 and linagliptin may exhibit synergistic effects in reducing IL-ip levels when used in combination compared to their individual effects. Table 29

[0419] In some aspects, mirodenafil AR1001 alone may reduce IL-ip levels in a concentration-dependent manner. For example, at 2 pM, mirodenafd ARI 001 may reduce IL- ip levels by 6.47%, while at 6 pM, the reduction may increase to 43.45%. Linagliptin alone at 1 pM may reduce IL-ip levels by 7.21%.

[0420] When mirodenafil AR 1001 and linagliptin are combined, the reduction in IL-ip levels may be greater than the sum of their individual effects. For instance, the combination of 2 pM mirodenafd AR1001 and 1 pM linagliptin may result in a 15.62% reduction in IL-ip levels. This combination may produce a synergistic effect, as indicated by an AB/(A+B) ratio of 1.14.

[0421] In some cases, increasing the concentration of mirodenafil AR 1001 in the combination may lead to an even greater reduction in IL-ip levels. The combination of 6 pM mirodenafd AR1001 and 1 pM linagliptin may result in a 60.50% reduction in IL-ip levels, with an AB/(A+B) ratio of 1.19, suggesting a stronger synergistic effect.

[0422] The synergistic effects observed in these combinations may indicate that mirodenafil AR 1001 and linagliptin may work through complementary mechanisms to reduce IL-ip levels in this inflammation model. These findings may suggest potential benefits of combining mirodenafil AR 1001 and linagliptin for managing inflammatory conditions associated with elevated IL-ip levels.

[0423] Referring to Table 30, the effects of combining mirodenafil AR1001 and linagliptin on TNFa levels in an LPS-induced inflammation model are presented. The data shows the TNFa fold change and reduction percentage for various concentrations of mirodenafil AR 1001 and linagliptin, both individually and in combination.

Table 30

[0424] In some aspects, mirodenafil AR1001 alone may reduce TNFa levels in a concentration-dependent manner. For example, at 2 pM, mirodenafil ARI 001 may reduce TNFa levels by 4.77%, while at 6 pM, the reduction may increase to 22.31%.

[0425] Linagliptin, when used alone at a concentration of 1 pM, may reduce TNFa levels by 11.33%. This reduction may be greater than that observed with the lower concentration of mirodenafil AR1001, but less than that seen with the higher concentration.

[0426] The combination of mirodenafil AR 1001 and linagliptin may result in a synergistic effect on TNFa reduction. When 2 pM mirodenafil AR1001 is combined with 1 pM linagliptin, the TNFa reduction may increase to 18.91%. This combination may produce a synergistic effect value of 1.17, as indicated by the AB/(A+B) ratio. [0427] In some cases, a more pronounced synergistic effect may be observed when using a higher concentration of mirodenafd ARI 001. The combination of 6 pM mirodenafd AR1001 with 1 pM linagliptin may result in a 40.58% reduction in TNFa levels. This combination may yield a synergistic effect value of 1.21, suggesting an enhanced cooperative effect between the two compounds at these concentrations.

[0428] The synergistic effect values greater than 1 for both combinations may indicate that the combined use of mirodenafil AR1001 and linagliptin may produce a greater reduction in TNFa levels than would be expected from the sum of their individual effects. This synergistic interaction may suggest a potential benefit in combining these compounds for reducing inflammatory responses in certain conditions.

[0429] EXAMPLES

[0430] Aspects of the present teachings may be further understood in light of the following examples, which should not be construed as limiting the scope of the present teachings in any way.

[0431 ] Example 1. Evaluation of Combination Therapy with Mirodenafil and Semaglutide

[0432] (1) Determine the effect of reducing intracellular Ap42

[0433] To investigate the synergistic effect on the reduction of intracellular Ap4 22 by the combination of mirodenafil and semaglutide, an Amyloid beta 42 Human ELISA was performed. Cells were treated with Ap at a final concentration of 5 pM, mirodenafil at a final concentration of 0.2, 0.6 pM, and semaglutide at a final concentration of 0.06, 0.14, 0.27 pM diluted in DMEM (1% FBS, 1% Penicillin/Streptomycin), incubated in an incubator at 37°C, 5% CO2 for 24 hours, and the experiment was performed as described in the Human Ap42 ELISA Kit, Ultrasensitive (invitrogen). First, the cell culture medium was removed, and then the cells were treated with 0.2 ml of RIPA lysis buffer, and the cells were crushed using a Cell Bioruptor for 15 minutes. Centrifuge at 14000 rpm, 4°C for 10 min, collect the supernatant, and add 50 pl of the lysate to each well of the Human Ap42 ELISA Kit (invitrogen) plate. Add 50 pl of Hu Ap42 Detection Antibody solution to each well and incubate for 3 hours at room temperature. Each well was emptied and the wells were washed with 200 pl of washing buffer (repeated 4 times). Add 100 pl of Anti-Rabbit IgGHRP solution to each well and leave at room temperature for 30 minutes, then empty each well and wash the wells with 200 pl of washing buffer (repeated 4 times). Add 100 P# of Stabilized Chromogen and incubate for 30 minutes at room temperature. Immediately after adding 100 pl of Stop Solution, the absorbance value was measured at 450 nm with a plate reader and the results are shown in Table 1 below. The experiment was repeated three times and statistical analysis was performed using Student's T-Test. All significance tests were performed at the P< 0.05 level:

[0434] In the following Tables, the combination index is as follows: (Combination Effect of A & B) / (AS ingle + B Single) = 1 : Additive Effect, > 1 : Synergic Effect.

Table 1

[0435] As shown in Table 1 above, the Ap reduction rate was significantly increased with the combination of mirodenafil and semaglutide compared to the Ap reduction rate with mirodenafil and semaglutide alone. Specifically, the Ap reduction rates of 0.547% and 0.699% for mirodenafil 0.2 pM and semaglutide 0.6 pM, respectively, were found to be significantly higher than the sum of the reduction rates of mirodenafil 0.2 pM and semaglutide 0.6 pM, but the Ap reduction rate of 3.573 was found to be significantly higher than the sum of the reduction rates of mirodenafil and semaglutide alone. Thus, the combination of mirodenafil and semaglutide demonstrated a synergistic effect in reducing Ap compared to mirodenafil and semaglutide alone.

[0436] (2) See if it reduces inflammatory factors

[0437] To investigate the synergistic effect of the combination of mirodenafil and semaglutide on the reduction of inflammatory factors, assays were performed using the Mouse ILip ELISA Kit and Mouse TNFa ELISA Kit. Mice were treated with LPS (10 ng/mL — 100 ng/mL), mirodenafil at concentrations of 2, 6 pM and semaglutide at concentrations of 0.6, 1.4, 2.8 pM, and incubated in a 37°C, 5% CO2 incubator for 24 hours. After 24 hours, the cultures of each group were collected in 15 ml conical tubes, and the Mouse IL-ip ELISA (Abeam, ab 197742) and Mouse TNF alpha ELISA Kit (Abeam, ab208348) were performed according to the method specified in the manual, respectively, and the results are shown in Tables 2 and 3 below.

Table 2

[0438] As shown in Table 2 above, the ILip reduction rate was significantly increased with the combination of mirodenafil and semaglutide compared to the ILip reduction rate with mirodenafil and semaglutide alone. Specifically, the ILip reduction rate of mirodenafil 2 pM and semaglutide 6 pM alone was 0.41% and 9.85%, respectively, but the ILip reduction rate of mirodenafil 2 pM and semaglutide 6 pM in combination was 14.52%, which was significantly higher than the sum of the reduction rates of mirodenafil and semaglutide alone. Thus, the combination of mirodenafil and semaglutide demonstrated a synergistic effect in reducing ILip compared to mirodenafil and semaglutide alone. Table 3

[0439] As can be seen from Table 3 above, TNFa reduction was significantly increased with the combination of mirodenafil and semaglutide compared to TNFa reduction with mirodenafil and semaglutide alone. Specifically, the TNFa reduction rate of mirodenafil 2 pM and semaglutide 6 pM alone was 6.94% and 8.15%, respectively, but the TNFa reduction rate of mirodenafil 2 pM and semaglutide 6 pM in combination was 16.17%, which was significantly higher than the sum of the reduction rates of mirodenafil and semaglutide alone. Thus, the combination of mirodenafil and semaglutide demonstrated a synergistic effect in reducing TNFa compared to mirodenafil and semaglutide alone.

[0440] Example 2. Evaluation of Combination Therapy with Mirodenafil and Sitagliptin

[0441 ] (1) Determine the effect of reducing intracellular Ap42

[0442] To investigate the synergistic effect of the combination of mirodenafil and sitagliptin on the reduction of intracellular Ap4 22, an Amyloid beta 42 Human ELISA was performed. Cells were treated with Ap diluted in DMEM (1% FBS, 1% Penicillin/Streptomycin) to a final concentration of 5 pM, mirodenafil to a final concentration of 0.2, 0.6 pM, and sitagliptin to a final concentration of 1, 2 pM, and incubated in a 37°C, 5% CO2 incubator for 24 hours. Then, as in Example 1 above, the experiment was performed according to the method specified in the Human Ap42 ELISA Kit, Ultrasensitive (invitrogen), and the results are shown in Table 4 below. Table 4

[0443] As shown in Table 4 above, the Ap reduction rate was significantly increased with the combination of mirodenafil and sitagliptin compared to the Ap reduction rate with mirodenafil and sitagliptin alone. Specifically, the Ap reduction rate of mirodenafil 0.2 pM and sitagliptin 1 pM alone was 2.36% and 0.75%, respectively, but the Ap reduction rate of mirodenafil 0.2 pM and sitagliptin 1 pM in combination was 5.83%, which was significantly higher than the sum of the reduction rates of mirodenafil and sitagliptin alone. Thus, the combination of mirodenafil and sitagliptin demonstrated a synergistic effect in reducing Ap compared to mirodenafil and sitagliptin alone.

[0444] (2) See if it reduces inflammatory factors

[0445] To determine the synergistic effect of the combination of mirodenafil and sitagliptin on the reduction of inflammatory factors, assays were performed using the Mouse ILip ELISA Kit and Mouse TNFa ELISA Kit using the same methods as in Example 1 above, and the results are shown in Tables 5 and 6 below.

Table 5

[0446] As shown in Table 5 above, the ILip reduction rate was significantly increased with the combination of mirodenafil and sitagliptin compared to the ILip reduction rate with mirodenafil and sitagliptin alone. Specifically, the ILip reduction rate of mirodenafil 2 pM and sitagliptin 10 pM alone was 8.07% and 12.23%, respectively, but the ILip reduction rate of mirodenafil 2 pM and sitagliptin 10 pM in combination was 23.54%, which was significantly higher than the sum of the reduction rates of mirodenafil and sitagliptin alone. Thus, the combination of mirodenafil and sitagliptin demonstrated a synergistic effect in reducing ILip compared to mirodenafil and sitagliptin alone.

Table 6

[0447] As shown in Table 6 above, the reduction in TNFa was significantly increased with the combination of mirodenafil and sitagliptin compared to the reduction in TNFa with mirodenafil and sitagliptin alone. Specifically, the TNFa reduction rate of mirodenafil 2 pM and sitagliptin 10 pM alone was 5.08% and 0.49%, respectively, but the TNFa reduction rate of mirodenafil 2 pM and sitagliptin 10 pM in combination was 6.64%, which was significantly higher than the sum of the reduction rates of mirodenafil and sitagliptin alone. Thus, the combination of mirodenafil and sitagliptin demonstrated a synergistic effect in reducing TNFa compared to mirodenafil and sitagliptin alone.

[0448] Example 3. Evaluation of Combination Therapy with Mirodenafil and Vildagliptin

[0449] (1) Determine the effect of reducing intracellular Ap42

[0450] To investigate the synergistic effect of the combination of mirodenafil and vildagliptin on the reduction of intracellular Ap4 22, an Amyloid beta 42 Human ELISA was performed. Cells were treated with Ap diluted in DMEM (1% FBS, 1% Penicillin/Streptomycin) to a final concentration of 5 pM, mirodenafil to a final concentration of 0.2, 0.6 pM, and vildagliptin to a final concentration of 1, 2 pM, and incubated in a 37°C, 5% CO2 incubator for 24 hours. Then, as in Example 1 above, the experiment was performed according to the method specified in Human Ap42 ELISA Kit, Ultrasensitive (invitrogen), and the results are shown in Table 7 below.

Table 7

[0451] As shown in Table 7 above, the Ap reduction was significantly increased with the combination of mirodenafil and vildagliptin compared to the Ap reduction with mirodenafil and vildagliptin alone. Specifically, the Ap reduction rate of mirodenafil 0.2 pM and vildagliptin 1 pM alone was 2.36% and 3.71%, respectively, but the Ap reduction rate of mirodenafil 0.2 pM and vildagliptin 1 pM in combination was 7.34%, which was significantly higher than the sum of the reduction rates of mirodenafil and vildagliptin alone. Thus, the combination of mirodenafil and vildagliptin demonstrated a synergistic effect on Ap reduction compared to mirodenafil and vildagliptin alone.

[0452] (2) See if it reduces inflammatory factors

[0453] To investigate the synergistic effect of the combination of mirodenafil and vildagliptin on the reduction of inflammatory factors, assays were performed using the Mouse ILip ELISA Kit and Mouse TNFa ELISA Kit using the same methods as in Example 1 above, and the results are shown in Tables 8 and 9 below.

Table 8

[0454] As can be seen from Table 8 above, there was a significant increase in ILip reduction with the combination of mirodenafil and vildagliptin compared to the ILip reduction with mirodenafil and vildagliptin alone. Specifically, the ILip reduction rate of mirodenafil 2 pM and vildagliptin 10 pM alone was 9.02% and 1.44%, respectively, but the ILip reduction rate of mirodenafil 2 pM and vildagliptin 10 pM in combination was 24.97%, which was significantly higher than the sum of the reduction rates of mirodenafil and vildagliptin alone. Thus, the combination of mirodenafil and vildagliptin demonstrated a synergistic effect in reducing ILip compared to mirodenafil and vildagliptin alone. Table 9

[0455] As shown in Table 9 above, the reduction in TNFa was significantly increased with the combination of mirodenafil and vildagliptin compared to the reduction in TNFa with mirodenafil and vildagliptin alone. Specifically, the TNFa reduction rate of mirodenafil 2 pM and vildagliptin 10 pM alone was 5.12% and 4.48%, respectively, but the TNFa reduction rate of mirodenafil 2 pM and vildagliptin 10 pM in combination was 11.19%, which was significantly higher than the sum of the reduction rates of mirodenafil and vildagliptin alone. Thus, the combination of mirodenafil and vildagliptin demonstrated a synergistic effect in reducing TNFa compared to mirodenafil and vildagliptin alone.

[0456] Example 4. Evaluation of Combination Therapy with Mirodenafil and Linagliptin

[0457] (1) Determine the effect of reducing intracellular Ap42

[0458] To determine the synergistic effect of the combination of mirodenafil and linagliptin on the reduction of intracellular Ap4 22, an Amyloid beta 42 Human ELISA was performed. Cells were treated with Ap diluted in DMEM (1% FBS, 1% Penicillin/Streptomycin) to a final concentration of 5 pM, mirodenafil to a final concentration of 0.2, 0.6 pM, and linagliptin to a final concentration of 0.1 pM, and incubated in a 37°C, 5% CO2 incubator for 24 hours. Then, as in Example 1 above, the experiment was performed according to the method specified in the Human Ap42 ELISA Kit, Ultrasensitive (invitrogen), and the results are shown in Table 10 below. Table 10

[0459] As shown in Table 10 above, the Ap reduction rate was significantly increased with the combination of mirodenafil and linagliptin compared to the Ap reduction rate with mirodenafil and linagliptin alone. Specifically, the Ap reduction rate of mirodenafil 0.2 pM and linagliptin 0.1 pM alone was 2.36% and 6.29%, respectively, but the Ap reduction rate of mirodenafil 0.2 pM and linagliptin 0.1 pM in combination was 15.73%, which was significantly higher than the sum of the reduction rates of mirodenafil and linagliptin alone. Thus, the combination of mirodenafil and linagliptin demonstrated a synergistic effect on Ap reduction compared to mirodenafil and linagliptin alone.

[0460] (2) See if it reduces inflammatory factors

[0461] To determine the synergistic effect of the combination of mirodenafil and linagliptin on the reduction of inflammatory factors, assays were performed using the Mouse ILip ELISA Kit and Mouse TNFa ELISA Kit using the same methods as in Example 1 above, and the results are shown in Tables 11 and 12 below.

Table 11

[0462] As can be seen from Table 11 above, there was a significant increase in ILip reduction with the combination of mirodenafil and linagliptin compared to the ILip reduction with mirodenafil and linagliptin alone. Specifically, the ILip reduction rate of mirodenafil 2 pM and linagliptin 1 pM alone was 96.47% and 7.21%, respectively, but the ILip reduction rate of mirodenafil 2 pM and linagliptin 1 pM in combination was 15.62%, which was significantly higher than the sum of the reduction rates of mirodenafil and linagliptin alone. Thus, the combination of mirodenafil and linagliptin demonstrated a synergistic effect in reducing ILip compared to mirodenafil and linagliptin alone.

Table 12

[0463] As shown in Table 12 above, the reduction in TNFa was significantly increased with the combination of mirodenafil and linagliptin compared to the reduction in TNFa with mirodenafil and linagliptin alone. Specifically, the TNFa reduction rate of 2 pM of mirodenafil and 1 pM of linagliptin alone was 4.77% and 11.33%, respectively, but the TNFa reduction rate of 2 pM of mirodenafil and 1 pM of linagliptin in combination was 18.92%, which was significantly higher than the sum of the reduction rates of mirodenafil and linagliptin alone. Thus, the combination of mirodenafil and linagliptin demonstrated a synergistic effect in reducing TNFa compared to mirodenafil and linagliptin alone.

[0464] Example 5

[0465] Culture method of HMC3 cells

[0466] The HMC3 cells, a human microglia cell line used in the experiments, were cultured, and maintained in DMED/F12 complete medium (HyClone) containing 10% fetal bovine serum (FBS, HyClone) and 1% antibiotic-antimycotic (GIBCO) at 37°C with 5% CO2 in a humidified CO2 incubator (Thermos fisher scientific). 2xl0³ cells were seeds in each 96- well plate and they were incubated for 24 hours in the humidified CO2 incubator as mentioned above. Further, LPS (lOOng/ml) or TNFa (5ng/ml) with/without the drugs, AR1001 and GLP-1 agonists (Semaglutide, Liraglutide, Tirzepatide, dulaglutide), were treated in the concentration of 0.001, 0.01, 0.1, 1 M individually or in combination.

[0467] Measurement of IL-6 by ELISA

[0468] IL-6 in culture supernatants were assayed by enzyme-linked immunosorbent assays (ELISA). The assays were conducted using Purified Rat Anti-Human IL-6, Biotin Rat Anti-Human IL-6 (BD Bioscience Pharmingen, CA, USA). IL-6 levels were derived from standard curves using the Thermo Scientific Skanlt Software (Thermos fisher scientific, Waltham, MA, USA).

[0469] Results of Measuring the pro-inflammatory cytokine (IL-6) in LPS-induced inflammation model

[0470] As can be seen from Figures 1-3, in LPS-induced inflammation, IL-6 levels were not significantly reduced when mirodenafil and GLP-1 agonists (Semaglutide, Liraglutide, Tirzepatide, dulaglutide) were administered alone, whereas IL-6 levels were significantly reduced when mirodenafil + semaglutide or mirodenafil + Tirzepatide or mirodenafil + dulaglutide were combined. However, there was no significant effect in combinations mirodenafil + Liraglutide.

[0471] As provided in Figure 1 and Table 1, there was no difference in the LPS- induced inflammatory response (IL-6) in a dose-dependent manner mirodenafil or semaglutide. However, when administered together, it was confirmed that mirodenafil 0.1 pM + semaglutide 0.1 pM (1 : 1) showed 16.67, mirodenafd 0.5 pM + semaglutide 0.1 pM (5: 1) showed 18.61, mirodenafil 1 pM + semaglutide 0.1 pM (10: 1) showed 14.20, and mirodenafil 5 pM + semaglutide 0.1 pM (50: 1) showed 12.26 reduction effects. Therefore, it was proven that when mirodenafil and semaglutide were administered together (1: 1 to 50: 1), there was a synergistic effect in reducing IL-6 compared to when they were administered alone.

Table 13

[0472] As provided in Figure 2 and Table 2, co-administration of high doses of mirodenafd and semaglutide was also effective. It was confirmed that mirodenafil 10 pM + semaglutide 0.1 pM (100: 1) showed 31.33, mirodenafil 10 pM + semaglutide 1 pM (10: 1) showed 36.31, mirodenafil 20 pM + semaglutide 0.1 pM (200: 1) showed 49.16, and mirodenafil 20 pM + semaglutide 1 pM (20: 1) showed 55.68 reduction effects. Therefore, it was proven that when mirodenafil and semaglutide were administered together (10: 1 to 200: 1), there was a synergistic effect in reducing IL-6 compared to when they were administered alone.

Table 14

[0473] As provided in Figure 3 and Table 3, here was no effect when low- concentration mirodenafil and tirzepatide were administered together, but a significant effect was observed when high-concentration mirodenafil and tirzepatide were administered together. It was confirmed that mirodenafil 10 pM + tirzepatide 0.1 pM (100: 1) showed 59.12, mirodenafil 10 pM + tirzepatide 1 pM (10: 1) showed 43.45, mirodenafil 20 pM + semaglutide 0.1 pM (200: 1) showed 51.87, and mirodenafil 20 pM + semaglutide 1 pM (20: 1) showed 89.63 reduction effects. Therefore, it was proven that when mirodenafil and semaglutide were administered together (10: 1 to 200: 1), there was a synergistic effect in reducing IL-6 compared to when they were administered alone. Table 15

[0474] As provided in Figure 4 and Table 4, there was no difference in the LPS- induced inflammatory response (IL-6) in a dose-dependent manner mirodenafil or dulaglutide. However, when administered together, it was confirmed that mirodenafil 0.1 pM + dulaglutide 0.001 pM (100: 1) showed 4.85, mirodenafil 0.1 pM + dulaglutide 0.01 pM (10: 1) showed 9.54, mirodenafil 0.1 pM + dulaglutide 0.1 pM (1 : 1) showed 9.31, mirodenafil 0.5 pM + dulaglutide 0.001 pM (500:1) showed 15.22, mirodenafil 0.5 pM + dulaglutide 0.01 pM (50: 1) showed 26.63, mirodenafil 1 pM + dulaglutide 0.001 pM (1000: 1) showed 22.16, mirodenafil 1 pM + dulaglutide 0.01 pM (100: 1) showed 16.42, and mirodenafil 1 pM + dulaglutide 0.1 pM (10: 1) showed 21.15 reduction effects. Therefore, it was proven that when mirodenafil and semaglutide were administered together (1: 1 to 1000: 1), there was a synergistic effect in reducing IL-6 compared to when they were administered alone. Table 16

Fold change LPS Treatment (% of Synergistic

(100 AR1001 Dulaglutide LPS AIL- 6 effect Rate ng/ml) (pM) (pM) group) ratio (AB/A+B) (A+B)

+ 100 0

A + .1 - 100.63 -0.63 + .5 - 92.51 7.49 + 1 - 91.79 8.21 + 5 - 93.21 6.79

B + .001 96.68 3.02 + .01 93.13 6.87 + .1 92.18 7.82 A+B + .1 .001 95.15 4.85 2.03 100: 1 + .1 .01 90.46 9.54 1.53 10: 1 + .1 .1 90.69 9.31 1.29 1: 1 + .5 .001 84.78 15.22 1.45 500: 1 + .5 .01 73.37 26.63 1.85 50: 1 + 1 .001 77.84 22.16 1.97 1000: 1 + 1 .01 83.58 16.42 1.09 100: 1 + 1 .1 78.85 21.15 1.32 10: 1

[0475] Results of Measuring the pro-inflammatory cytokine (IL-6) in TNFa- induced inflammation model

[0476] As can be seen from [Figures 5-6], in TNFa- induced inflammation, IL-6 levels were not significantly reduced when mirodenafil and GLP-1 agonists (Semaglutide, Liraglutide, Tirzepatide, dulaglutide) were administered alone, whereas IL-6 levels were significantly reduced when mirodenafil + semaglutide or mirodenafil + Tirzepatide or mirodenafil + dulaglutide were combined. However, there was no significant effect in combinations mirodenafil + Liraglutide.

[0477] As provided in Figure 5 and Table 5, there was no effect when low- concentration mirodenafil and semaglutide were administered together, but a significant effect was observed when high-concentration mirodenafil and semaglutide were administered together. It was confirmed that mirodenafil 10 pM + semaglutide 0.01 pM (1000: 1) showed 23.70, mirodenafil 10 pM + semaglutide 0.1 pM (100: 1) showed 25.34, mirodenafil 10 pM + semaglutide 1 pM (10: 1) showed 27.54, mirodenafil 20 pM + semaglutide 0.1 pM (200: 1) showed 24.68, and mirodenafil 20 pM + semaglutide 1 pM (20: 1) showed 41.58 reduction effects. Therefore, it was proven that when mirodenafil and semaglutide were administered together (10: 1 to 1000: 1), there was a synergistic effect in reducing IL-6 compared to when they were administered alone.

Table 17

[0478] As provided in Figure 6 and Table 6, there was no difference in the TNFa- induced inflammatory response (IL-6) in a dose-dependent manner mirodenafil or Tirzepatide. However, when administered together, it was confirmed that mirodenafil 0.1 pM + Tirzepatide 0.001 pM (100: 1) showed 20.51, mirodenafil 0.1 pM + Tirzepatide 0.01 pM (10: 1) showed 13.27, mirodenafil 0.5 pM + Tirzepatide 0.1 pM (5: 1) showed 11.19, mirodenafil 1 pM + Tirzepatide 0.1 pM (10: 1) showed 14.54, mirodenafil 10 pM + Tirzepatide 1 pM (10: 1) showed 19.26, mirodenafil 20 pM + Tirzepatide 0.1 pM (200: 1) showed 15.16, and mirodenafil 20 pM + Tirzepatide 1 pM (20: 1) showed 36.01 reduction effects. Therefore, it was proven that when mirodenafil and Tirzepatide were administered together (1:1 to 200: 1), there was a synergistic effect in reducing IL-6 compared to when they were administered alone.

Table 18

[0479] As provided in Figure 7 and Table 7, there was no difference in the TNFa- induced inflammatory response (IL-6) in a dose-dependent manner mirodenafil or dulaglutide. However, when administered together, it was confirmed that mirodenafil 0.1 pM + dulaglutide 0.001 pM (100: 1) showed 12.05, mirodenafil 0.1 pM + dulaglutide 0.01 pM (10: 1) showed 19, mirodenafil 0.1 pM + dulaglutide 0.1 pM (1: 1) showed 12.64, mirodenafil 0.5 pM + dulaglutide 0.001 pM (500:1) showed 13.76, mirodenafil 0.5 pM + dulaglutide 0.01 pM (50: 1) showed 15.31, mirodenafil 0.5 pM + dulaglutide 0.1 pM (5: 1) showed 17.74, mirodenafil 1 pM + dulaglutide 0.01 pM (100: 1) showed 14.12, and mirodenafil 1 pM + dulaglutide 0.1 pM (10: 1) showed 8.16 reduction effects. Therefore, it was proven that when mirodenafil and semaglutide were administered together (1: 1 to 500: 1), there was a synergistic effect in reducing IL-6 compared to when they were administered alone.

Table 19

Fold change

LPS Treatment (% of Synergistic

(100 AR1001 Dulaglutide LPS AIL-6 effect Rate ng/ml) (pM) (pM) group) ratio (AB/A+B) (A+B)

+ - - 100 0

A + .1 - 100.2 -.02

+ .5 - 98.25 1.75 + 1 91A1 2.53

B + .001 97.47 2.53

+ .01 97.03 2.97

+ .1 97.93 2.07

A+B + .1 .001 87.95 12.05 4.80 100: 1

+ .1 .01 81 19 6.44 10: 1

+ .1 .1 87.36 12.64 6.17 1: 1

+ .5 .001 86.24 13.76 3.21 500: 1

+ .5 .01 84.69 15.31 3.24 50: 1

+ .5 .1 82.26 17.74 4.64 5: 1

+ 1 .01 85.88 14.12 2.57 100: 1

+ 1 .1 91.84 8.16 1.77 10: 1

[0480] Example 6

[0481] Culture method of primary astrocytes

[0482] Primary cultures of mouse cortical neurons were prepared from the pregnant mouse (TP14.5-15.5). Briefly, cortices free of meninges were dissociated in 0.125% (w/v) trypsin at 37°C. The tissue fragments were dissociated mechanically by repeated gentle pipetting through wide and narrow-bore fire-polished Pasteur pipettes in culture medium [DMEM/F12 (Hyclone) containing 10% fetal bovine serum (FBS, HyClone) and 1% antibiotic-antimycotic (GIBCO) at 37°C with 5% CO2 in a humidified CO2 incubator (Thermos fisher scientific). Cells were counted and then plated in multiwells (96 wells; SPL, Korea) at a density of 30,000 cells per well. Cultures were grown at 37°C in humidified atmosphere, 5% CO2/95% air. The cultures were used in experiments after 7 days of in vitro incubation (days in vitro: DIV).

[0483] Measurement of IL-6 by ELISA

[0484] IL-6 in culture supernatants were assayed by enzyme-linked immunosorbent assays (ELISA). The assays were conducted using Purified Rat Anti-Human IL-6, Biotin Rat Anti-Human IL-6 (BD Bioscience Pharmingen, CA, USA). IL-6 levels were derived from standard curves using the Thermo Scientific Skanlt Software (Thermos fisher scientific, Waltham, MA, USA).

[0485] Results of Measuring the pro-inflammatory cytokine (IL-6) in A-beta + IFNr or STZ -induced inflammation model

[0486] As provided in Figure 8 and Tables 8-9, in primary astrocytes, IL-6, an inflammatory marker, was confirmed after treatment with A-beta (10pM/ml)+IFNr (20ng/ml) or STZ (lOmM/ml). When administered together, it was confirmed that mirodenafil 1 pM + Exenatide 0.1 pM (10: 1) showed 22.19 and mirodenafil 1 pM + Exenatide 1 pM (11 : 1) showed 11.45 reduction effects. Therefore, it was proven that when mirodenafil and Exenatide were administered together (1: 1 to 10:1), there was a synergistic effect in reducing IL-6 compared to when they were administered alone. As with A-beta+IFNr, IL-6 was also decreased when mirodenafil+exenatide was co-treated with STZ.

Table 20

Table 21

[0487] Example 3

[0488] Culture method of primary neuron culture

[0489] Primary cultures of mouse cortical neurons were prepared from the pregnant mouse (TP14.5-15.5). Briefly, cortices free of meninges were dissociated in 0.125% (w/v) trypsin at 37°C. The tissue fragments were dissociated mechanically by repeated gentle pipetting through wide and narrow-bore fire-polished Pasteur pipettes in culture medium [Neurobasal medium (Gibco, Grand Island, NY, USA) supplemented with 2% B27® Supplement (50X), serum free (Gibco®)]. Cells were counted and then plated in a poly-L- lysine (5 g/ml)-coated multiwells (96 wells; SPL, Korea) at a density of 30,000 cells per well. Cultures were grown at 37°C in humidified atmosphere, 5% CO2/95% air. The cultures were used in experiments after 7 days of in vitro incubation (days in vitro: DIV).

[0490] Mitochondrial membrane potential (JC-1 assay)

[0491] Following 48 hours of incubation at 37°C with 5% CO2, the mitochondrial membrane potential was assessed using the JC-1 assay. The JC-1 Mitochondrial Membrane Potential Assay Kit (abl 13850, Abeam) was used according to the manufacturer’s protocol, with the cells stained for 10 minutes in a CO2 incubator. Fluorescent measurements were taken using a microplate reader (Varioskan LUX), with results expressed as the ratio of aggregate (535 nm, red) to monomer (475 nm, green).

[0492] Results of Measuring JC-1 ratio in A-beta induced in primary neuron

[0493] As provided in Figure 9 and Table 10, the decrease in JC-1 ratio (Aggregate/Monomer) in primary neurons following Ap treatment was compared to the JC-1 ratio changes in the drug-treated groups. There was no difference in the Ap-induced mitochondrial damage (JC-1) in a dose-dependent manner mirodenafil or semaglutide. However, when administered together, it was confirmed that mirodenafil 0.1 pM + semaglutide 0.01 pM (10: 1) showed 28, mirodenafil 0.1 pM + dulaglutide 0.1 pM (1 : 1) showed 18, mirodenafil 0.5 pM + dulaglutide 0.01 pM (50: 1) showed 6, mirodenafil 0.5 pM + dulaglutide 0.01 pM (50: 1) showed 14, mirodenafil 1 pM + dulaglutide 0.01 pM (100: 1) showed 14, and mirodenafil 5 pM + dulaglutide 1 pM (5: 1) showed 21 increase effects. Therefore, it was proven that when mirodenafil and semaglutide were administered together (1: 1 to 100: 1), there was a synergistic effect in inducing JC-1 compared to when they were administered alone. Table 22

As provided in Figure 10 and Table 11, the decrease in JC-1 ratio (Aggregate/Monomer) in primary neurons following Ap treatment was compared to the JC-1 ratio changes in the drug-treated groups. There was no difference in the Ap-induced mitochondrial damage (JC-1) in a dose-dependent manner mirodenafil or Exenatide. However, when administered together, it was confirmed that mirodenafil 0.1 pM + Exenatide 0.001 pM (100: 1) showed 6, and mirodenafil 0.5 pM + dulaglutide 0.001 pM (500: 1) showed 15 increase effects. Therefore, it was proven that when mirodenafil and exenatide were administered together (1: 1 to 100: 1), there was a synergistic effect in inducing JC-1 compared to when they were administered alone. Table 23

[0494] Statistical analysis

[0495] The values are indicated as Mean ± SD and statistical significance was analyzed by One Way ANOVA.

[0496] Other Embodiments

[0497] The detailed description set-forth above is provided to aid those skilled in the art in practicing the present invention. However, the invention described and claimed herein is not to be limited in scope by the specific embodiments herein disclosed because these embodiments are intended as illustration of several aspects of the invention. Any equivalent embodiments are intended to be within the scope of this invention. Indeed, various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description which do not depart from the spirit or scope of the present inventive discovery. Such modifications are also intended to fall within the scope of the appended claims.

[0498] References Cited

[0499] All publications, patents, patent applications and other references cited in this application are incorporated herein by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent application or other reference was specifically and individually indicated to be incorporated by reference in its entirety for all purposes. Citation of a reference herein shall not be construed as an admission that such is prior art to the present invention.

Claims

CLAIMS What is claimed is:

1. A pharmaceutical composition for the prevention or treatment of neurodegenerative disorders, comprising: a PDE-5 inhibitor; and a GLP-1 receptor agonist, a GIP agonist, and and glucagon wherein the molar ratio of any individual member of the group above to another member is between about (1,0,0,1000) to about (1000,0,0,0).

2. A pharmaceutical composition for the prevention or treatment of neurodegenerative disorders, comprising: a PDE-5 inhibitor; and a DPP-4 inhibitor, a GIP agonist, and and glucagon wherein the molar ratio of any individual member of the group above to another member is between about (1,0,0,1000) to about (1000,0,0,0).

3. A pharmaceutical composition for the prevention or treatment of neurodegenerative disorders, comprising: a PDE-5 inhibitor; and a GLP-1 receptor agonist, wherein the molar ratio of any individual member of the group above to another member is between about (1,0,0,1000) to about (1000,0,0,0).

4. A pharmaceutical composition for the prevention or treatment of neurodegenerative disorders, comprising: a PDE-5 inhibitor; and a dual GLP- 1 receptor agonist and GIP agonist, wherein the molar ratio of any individual member of the group above to another member is between about (1,0,0,1000) to about (1000,0,0,0).

5. A pharmaceutical composition for the prevention or treatment of neurodegenerative disorders, comprising: a PDE-5 inhibitor; and a triple GLP-1 receptor agonist, GIP agonist, and glucagon agonist, wherein the molar ratio of any individual member of the group above to another member is between about (1,0,0,1000) to about (1000,0,0,0).

6. The pharmaceutical composition of claims 1 through 5, wherein the PDE-5 inhibitor is selected from the group consisting of mirodenafil, sildenafil, vardenafil, tadalafil, udenafil, dasantafil, avanafil, and pharmaceutically acceptable salts, solvates and hydrates thereof.

7. The pharmaceutical composition of claims 1 through 5, wherein the GLP-1 receptor agonist is selected from the group consisting of semaglutide, exenatide, liraglutide, lixisenatide, albiglutide, dulaglutide, tirzepatide, cotadutide, and taspoglutide.

8. The pharmaceutical composition of claim 6, wherein the PDE-5 inhibitor is mirodenafil and pharmaceutically acceptable salts thereof.

9. The pharmaceutical composition of claim 7, wherein the GLP-1 receptor agonist is semaglutide.

10. The pharmaceutical composition of claim 2, wherein the DPP-4 inhibitor is sitagliptin.

11. The pharmaceutical composition of claims 1 through 5, wherein the neurodegenerative disorder is selected from the group consisting of Parkinson's disease (PD); sporadic or heritable dementia with Lewy bodies (DLB); pure autonomic failure (PAF) with aS deposition; multiple system atrophy (MSA); hereditary neurodegeneration with brain iron accumulation; and incidental Lewy body disease of advanced age; Alzheimer's disease of the Lewy body variant; Down's syndrome; progressive supranuclear palsy; essential tremor with Lewy bodies; familial parkinsonism with or without dementia resulting from a mutant gene and loci where no gene mutation has yet been identified; Creutzfeldt Jakob disease; bovine spongiform encephalopathy; secondary Parkinson disease/parkinsonism resulting from neurotoxin exposure/drug-induced parkinsonism with a-synuclein deposition; sporadic or heritable spinocerebellar ataxia; amyotrophic lateral sclerosis (ALS); idiopathic rapid eye movement sleep behavior disorder; and other conditions associated with central and/or peripheral a-synuclein accumulation in mammals accompanying a primary disease process.

12. The pharmaceutical composition of claims 1 through 5, wherein the neurodegenerative disorder is selected from the group consisting of Alzheimer’s disease, Vascular dementia, Frontotemporal dementia (FTD), Lewy body dementia (LBD), Mixed dementia, Posterior cortical atrophy (PCA), Primary progressive aphasia (PPA), Corticobasal degeneration (CBD), and Progressive supranuclear palsy (PSP), idiopathic myeloma, amyloid polyneuropathy, amyloid cardiomyopathy, systemic senile amyloidosis, amyloid polyneuropathy, hereditary cerebral hemorrhage with amyloidosis, Down's syndrome, Scrapie, medullary carcinoma of the thyroid, isolated atrial amyloidosis, P2- microglobulin amyloidosis, inclusion body myositis, muscle wasting disease, Islets of Langerhans diabetes, Type 1 diabetes, insulinoma, Type 2 diabetes mellitus, hereditary cerebral hemorrhage amyloidosis (Dutch), amyloid A (reactive) amyloidosis, secondary amyloidosis, familial Mediterranean fever, familial amyloid nephropathy with urticaria and deafness (Muckle-wells Syndrome), amyloid lambda L-chain amyloidosis, amyloid kappa L-chain amyloidosis, idiopathic associated amyloidosis, myeloma- associated amyloidosis, macroglobulinemia-associated amyloidosis, A beta 2M amyloidosis (chronic hemodialysis), ATTR amyloidosis (familial amyloid polyneuropathy (Portuguese, Japanese, Swedish)), familial amyloid cardiomyopathy (Danish), isolated cardiac amyloidosis, systemic senile amyloidosis, AIAPP or amylin insulinoma, atrial natriuretic factor amyloidosis (isolated atrial amyloidosis), procalcitonin amyloidosis (medullary carcinoma of the thyroid), gelsolin amyloidosis (familial amyloidosis (Finnish)), cystatin C (hereditary cerebral hemorrhage with amyloidosis (Icelandic)), AApo-A-I amyloidosis (familial amyloidotic polyneuropathy-Iowa), AApo-A-II amyloidosis, traumatic brain injury, fibrinogen- associated amyloidosis, Creutzfeldt- Jakob disease, Gertsmann-Straussler-Scheinker syndrome, bovine spongiform encephalitis, condition associated with homozygosity for the apolipoprotein E4 allele, and Huntington's disease.

13. The pharmaceutical composition of claims 1 through 5, wherein the neurodegenerative disorder is selected from the group consisting of Parkinson's Disease (PD), Parkinson's Disease Dementia (PDD), Dementia with Lewy Bodies (DLB), Multiple System Atrophy (MSA), Pure Autonomic Failure (PAF), and Lewy Body Variant of Alzheimer's Disease (LBV).

14. The pharmaceutical composition of claims 1 through 5, wherein the neurodegenerative disorder is selected from the group consisting of Alzheimer’ s disease, Vascular dementia, Frontotemporal dementia (FTD), Lewy body dementia (LBD), Mixed dementia, Posterior cortical atrophy (PCA), Primary progressive aphasia (PPA), Corticobasal degeneration (CBD), and Progressive supranuclear palsy (PSP).

15. The pharmaceutical composition of claims 1 through 5, wherein the neurodegenerative disorder is selected from the group consisting of Dementia with Lewy

Bodies and Alzheimer’s disease.

16. The pharmaceutical composition of claims 1 through 5, wherein the pharmaceutical composition is formulated for oral administration.

17. The pharmaceutical composition of claim 16, wherein the pharmaceutical composition is in the form of a tablet or capsule.

18. A method for treating a neurodegenerative disorder in a subject in need thereof, the method comprising: administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a PDE-5 inhibitor, a GLP-1 receptor agonist, a GIP agonist, and glucagon, wherein the molar ratio of any individual member of the group above to another member is between about (1,0,0,1000) to about (1000,0,0,0).

19. A method for treating a neurodegenerative disorder in a subject in need thereof, the method comprising: administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a PDE-5 inhibitor, a DPP-4 inhibitor, a GIP agonist, and glucagon, wherein the molar ratio of any individual member of the group above to another member is between about (1,0,0,1000) to about (1000,0,0,0).

20. A method for treating a neurodegenerative disorder in a subject in need thereof, the method comprising: administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a PDE-5 inhibitor, a GLP-1 receptor and GIP dual agonist, wherein the molar ratio of any individual member of the group above to another member is between about (1,0,0,1000) to about (1000,0,0,0).

21. A method for treating a neurodegenerative disorder in a subject in need thereof, the method comprising: administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a PDE-5 inhibitor, a GLP-1 receptor, GIP, and glucagon triple agonist, wherein the molar ratio of any individual member of the group above to another member is between about (1,0,0,1000) to about (1000,0,0,0).

22. The method of claims 18 through 21, wherein the PDE-5 inhibitor is selected from the group consisting of mirodenafil, sildenafil, vardenafil, tadalafil, udenafil, dasantafil, avanafil, and pharmaceutically acceptable salts, solvates and hydrates thereof.

23. The method of claims 18 through 21, wherein the PDE-5 inhibitor is mirodenafil.

24. The method of claim 19, wherein the DPP-4 inhibitor is selected from the group consisting of sitagliptin, vildagliptin, saxagliptin, linagliptin, gemigliptin, teneligliptin, alogliptin, evogliptin, and anagliptin.

25. The method of claim 24, wherein the DPP-4 inhibitor is sitagliptin.

26. The method of claims 18 through 21, wherein the neurodegenerative disorder is selected from the group consisting of Parkinson's disease (PD); sporadic or heritable dementia with Lewy bodies (DLB); pure autonomic failure (PAF) with aS deposition; multiple system atrophy (MSA); hereditary neurodegeneration with brain iron accumulation; and incidental Lewy body disease of advanced age; Alzheimer's disease of the Lewy body variant; Down's syndrome; progressive supranuclear palsy; essential tremor with Lewy bodies; familial parkinsonism with or without dementia resulting from a mutant gene and loci where no gene mutation has yet been identified; Creutzfeldt Jakob disease; bovine spongiform encephalopathy; secondary Parkinson disease/parkinsonism resulting from neurotoxin exposure/drug-induced parkinsonism with a-synuclein deposition; sporadic or heritable spinocerebellar ataxia; amyotrophic lateral sclerosis (ALS); idiopathic rapid eye movement sleep behavior disorder; and other conditions associated with central and/or peripheral a-synuclein accumulation in mammals accompanying a primary disease process.

27. The method of claims 18 through 21, wherein the neurodegenerative disorder is selected from the group consisting of Alzheimer’s disease, Vascular dementia, Frontotemporal dementia (FTD), Lewy body dementia (LBD), Mixed dementia, Posterior cortical atrophy (PCA), Primary progressive aphasia (PPA), Corticobasal degeneration (CBD), and Progressive supranuclear palsy (PSP), idiopathic myeloma, amyloid polyneuropathy, amyloid cardiomyopathy, systemic senile amyloidosis, amyloid polyneuropathy, hereditary cerebral hemorrhage with amyloidosis, Down's syndrome, Scrapie, medullary carcinoma of the thyroid, isolated atrial amyloidosis, P2- microglobulin amyloidosis, inclusion body myositis, muscle wasting disease, Islets of Langerhans diabetes, Type 1 diabetes, insulinoma, Type 2 diabetes mellitus, hereditary cerebral hemorrhage amyloidosis (Dutch), amyloid A (reactive) amyloidosis, secondary amyloidosis, familial Mediterranean fever, familial amyloid nephropathy with urticaria and deafness (Muckle-wells Syndrome), amyloid lambda L-chain amyloidosis, amyloid kappa L-chain amyloidosis, idiopathic associated amyloidosis, myeloma- associated amyloidosis, macroglobulinemia-associated amyloidosis, A beta 2M amyloidosis (chronic hemodialysis), ATTR amyloidosis (familial amyloid polyneuropathy (Portuguese, Japanese, Swedish)), familial amyloid cardiomyopathy (Danish), isolated cardiac amyloidosis, systemic senile amyloidosis, AIAPP or amylin insulinoma, atrial natriuretic factor amyloidosis (isolated atrial amyloidosis), procalcitonin amyloidosis (medullary carcinoma of the thyroid), gelsolin amyloidosis (familial amyloidosis (Finnish)), cystatin C (hereditary cerebral hemorrhage with amyloidosis (Icelandic)), AApo-A-I amyloidosis (familial amyloidotic polyneuropathy-Iowa), AApo-A-II amyloidosis, traumatic brain injury, fibrinogen- associated amyloidosis, Creutzfeldt- Jakob disease, Gertsmann-Straussler-Scheinker syndrome, bovine spongiform encephalitis, condition associated with homozygosity for the apolipoprotein E4 allele, and Huntington's disease.

28. The method of claims 18 through 21, wherein the neurodegenerative disorder is selected from the group consisting of Parkinson's Disease (PD), Parkinson's Disease Dementia (PDD), Dementia with Lewy Bodies (DLB), Multiple System Atrophy (MSA), Pure Autonomic Failure (PAF), and Lewy Body Variant of Alzheimer's Disease (LBV).

29. The method of claims 18 through 21, wherein the neurodegenerative disorder is selected from the group consisting of Alzheimer’s disease, Vascular dementia, Frontotemporal dementia (FTD), Lewy body dementia (LBD), Mixed dementia, Posterior cortical atrophy (PCA), Primary progressive aphasia (PPA), Corticobasal degeneration (CBD), and Progressive supranuclear palsy (PSP).

30. The method of claims 18 through 21, wherein the neurodegenerative disorder is selected from the group consisting of Dementia with Lewy Bodies and Alzheimer’s disease.