Dual inhibitors of amine oxidases and peroxidases, and uses thereof

Abstract

The present invention relates to novel compounds which are capable of inhibiting semicarbazide-sensitive amine oxidase and myeloperoxidase. These compounds are useful for the treatment of a variety of inflammatory indications, e.g., autoimmune diseases, respiratory disease, fibrosis, vasculitis, neurological diseases or disorders, and the complications resulting from infectious disease, obesity, diabetes / metabolic syndrome and / or cardiovascular disease in human subjects as well as in pets and livestock. In addition, the present invention relates to pharmaceutical compositions containing these compounds, as well as various uses thereof.

Description

DUAL INHIBITORS OF AMINE OXIDASES AND PEROXIDASES, AND USES THEREOF Cross Reference

[0001] The present application claims priority to Australian provisional application number 2023903784 filed on 24 November 2023, the entire contents of which is herein incorporated by cross reference in its entirety. Technical Field

[0002] The present invention relates to novel compounds which are capable of inhibiting semicarbazide- sensitive amine oxidase and myeloperoxidase. These compounds are useful for the treatment of a variety of inflammatory indications, e.g., autoimmune diseases, respiratory disease, fibrosis, vasculitis, neurological diseases or disorders, and the complications resulting from infectious disease, obesity, diabetes / metabolic syndrome and / or cardiovascular disease in human subjects as well as in pets and livestock. In addition, the present invention relates to pharmaceutical compositions containing these compounds, as well as various uses thereof. Background

[0003] Inflammatory diseases encompass a wide range of conditions characterized by an abnormal immune response, leading to chronic inflammation of various tissues and organs in the body. This dysregulated immune response can result in tissue damage, pain, and functional impairment, affecting millions of individuals worldwide. These conditions include autoimmune disease, respiratory disease, fibrosis, neurological disease, and many others. The pathology of inflammatory diseases involves the activation of immune cells, particularly neutrophils and macrophages, which release inflammatory mediators such as cytokines, chemokines, and reactive oxygen species. This increased inflammatory response attracts more immune cells to the affected site, perpetuating the cycle of chronic inflammation. Over time, this sustained inflammation can lead to tissue destruction and organ dysfunction. The global health impact of inflammatory diseases is significant, both in terms of financial burden and quality of life for affected individuals. According to the World Health Organization, chronic inflammatory diseases are among the leading causes of morbidity and mortality worldwide. They can have profound physical, psychological, and social consequences, limiting mobility, impairing daily activities, and reducing overall well-being. Furthermore, the economic impact of these diseases is substantial, encompassing direct healthcare costs, lost productivity, and decreased quality of life. The burden is particularly high in developed countries where the prevalence of inflammatory diseases is rising due to factors such as agingpopulations, sedentary lifestyles, and environmental triggers. The development of effective anti- inflammatory drugs plays a crucial role in managing these diseases and alleviating symptoms. By targeting specific molecular pathways involved in the inflammatory response, anti-inflammatory drugs can help modulate and regulate the immune response, reducing inflammation and preventing tissue damage. These medications provide relief to individuals suffering from inflammatory diseases, improving their quality of life and potentially reducing long-term complications.

[0004] SSAO / VAP-1 (semicarbazide sensitive amine oxidase / vascular adhesion protein 1) and MPO (myeloperoxidase) are enzymes that play a critical role in inflammation. SSAO / VAP-1 is an enzyme that catalyzes the oxidation of primary amines, such as amino acids and biogenic amines. In the context of inflammation, SSAO / VAP-1 has been found to be involved in various processes, including the generation of reactive oxygen species (ROS) and the regulation of vascular functions. ROS, produced by SSAO / VAP- 1, can contribute to the oxidative stress and tissue damage associated with inflammation. Additionally, SSAO / VAP-1 has been shown to influence the migration and adhesion of immune cells, further contributing to the inflammatory response. MPO, on the other hand, is an enzyme primarily found in neutrophils, a type of white blood cell. MPO plays a crucial role in the immune system's defence against pathogens. It generates hypochlorous acid and other reactive intermediates, which have antimicrobial properties. However, excessive or dysregulated MPO activity leads to tissue damage and inflammation. MPO has been implicated in various inflammatory diseases, including atherosclerosis, rheumatoid arthritis, and chronic obstructive pulmonary disease (COPD) to name a few. Its presence and activity in inflamed tissues contribute to the production of oxidative and nitrosative stress, promoting further inflammation. Both SSAO / VAP-1 and MPO have been studied extensively for their involvement in inflammation, and targeting these enzymes may potentially offer therapeutic benefits for managing inflammatory diseases.

[0005] In most organisms, including humans, two families of mammalian amine oxidases metabolize various mono-, di-, and polyamines produced endogenously or absorbed from exogenous sources. The first family includes the monoamine oxidases (MAO-A and MAO-B) which are present in the mitochondria of most cell types and use covalently bound flavin adenine dinucleotide (FAD) as the cofactor. Polyamine oxidase is another FAD-dependent amine oxidase which oxidatively deaminates spermine and spermidine. The second family is dependent on copper and uses other co-factors apart from FAD, such as an oxidized tyrosine residue (abbreviated as TPQ or LTQ). One member of this family is semicarbazide-sensitive amine oxidase (SSAO), also known as primary amine oxidase, plasma amine oxidase and benzylamine oxidase, and is identical to vascular adhesion protein-1 (VAP-1 – herein described as SSAO / VAP-1). Diamine oxidase (DAO) and the lysyl oxidase family of proteins (lysyl oxidase (LOX) and lysyl oxidase-like (LOXL) 1-4) also belong to this second family. Pharmacologically, MAO-A is selectively inhibited by clorgyline, MAO-B by L-deprenyl and SSAO / VAP-1 by semicarbazide.

[0006] SSAO / VAP-1 is an ectoenzyme containing a very short cytoplasmic tail, a single transmembrane domain, and a large, highly glycosylated extracellular domain which contains the active site for the amine oxidase activity. SSAO / VAP-1 is also present in a soluble form circulating in the plasma of some animals. It has been shown that this form is a cleaved product of membrane-bound SSAO / VAP-1. SSAO / VAP-1 is highly expressed in adipose tissue, lung, liver, ileum and aorta, and the roles it plays in each organ have only recently begun to be investigated.

[0007] Oxidative deamination of primary monoamines by SSAO / VAP-1 produces ammonia (NH3), aldehydes and hydrogen peroxide (H2O2), agents with established toxicity that can cause significant inflammation through their promiscuous reactivity and oxidative stress. During the SSAO / VAP-1 amine oxidase catalytic cycle the covalently bound cofactor, TPQ, is first reduced, and then re-oxidized by oxygen in the presence of copper with the generation of ammonia and hydrogen peroxide as by-products. It has been speculated that excessive hydrogen peroxide concentrations can be deleterious and may contribute to the pathology of various inflammatory and neurodegenerative processes.

[0008] The role of SSAO / VAP-1 in inflammatory diseases has been reviewed. SSAO / VAP-1 appears to have two physiological functions: the first is the amine oxidase activity and the second is cell adhesion activity. Both activities are associated with inflammatory processes. Inhibitors of the amine oxidase activity of SSAO / VAP-1 have been found to interfere with leukocyte rolling, adhesion and extravasation and, similar to SSAO / VAP-1 antibodies, exhibit anti-inflammatory properties. Excessive and chronic inflammatory responses have been associated with the symptoms of many chronic diseases, such as rheumatoid arthritis, multiple sclerosis and asthma. Patients suffering from either atopic eczema or psoriasis (both chronic inflammatory skin disorders) have higher levels of SSAO / VAP-1 positive cells in their skin compared to skin from healthy controls.

[0009] It has been shown that SSAO / VAP-1 blocking by enzyme inhibitors has a pronounced effect in neutrophil-driven animal models of inflammation. Unregulated neutrophil activation and activity can lead to release of excessive MPO which reacts with hydrogen peroxide to form hypochlorous acid that causes local tissue damage and further inflammation.

[0010] MPO is a highly cationic, heme-containing, glycosylated enzyme which is found mainly in neutrophils, making up 5% of the total dry weight of these leukocytic cells. It is also found to a lesser degree in macrophages and monocytes. It belongs to a large family of mammalian peroxidases that also includes the closely related eosinophil peroxidase (EPO), thyroid peroxidase (TPO) lactoperoxidase (LPO) and vascular peroxidase (VPO). The mature form of this protein is a homo-dimer linked through a disulphide bridge, with each monomer containing a covalently bound heme cofactor that is responsible for the highly characteristic green colour of MPO. Mature MPO is stored in azurophilic granules of fully differentiatedneutrophils in a dormant state. Following priming and activation by a variety of inflammatory mediators, MPO can be released from the neutrophils by rapid degranulation and acts on the immediate environment. Under certain conditions, MPO can also be released by neutrophils via the extrusion of neutrophil extracellular traps (NETs) allowing for extracellular, remote targeting of pathogens, and in response to specific inflammatory stimuli.

[0011] In the presence of hydrogen peroxide, the released MPO is activated and becomes a potent oxidant able to oxidize a variety of physiological low molecular weight entities (Cl-, Br-, SCN- or tyrosine) forming highly reactive species such as HOCl, HOBr, HOSCN and the tyrosyl radical. The hydrogen peroxide required for MPO function can come from a variety of sources, such as from superoxide dismutase (SOD) turnover of the superoxide anion during respiratory burst or from amine oxidases, such as SSAO / VAP-1, during metabolism of their endogenous substrates.

[0012] Due to the high concentrations of chloride present in most biological fluids, it is thought that this is the most relevant of the physiological substrates, forming hypochlorous acid (HOCl, bleach) capable of reacting with a variety of targets such as amino acids (isolated and as part of a protein), DNA and lipids rapidly and indiscriminately, significantly affecting cellular function. Hypochlorous acid readily chlorinates tyrosine to 3-chlorotyrosine which is used as a specific biomarker for MPO activity resulting in HOCl production in vivo.

[0013] Asthma is a disease resulting from chronic inflammation of the airways ensuing in bronchoconstriction and excessive build-up of mucus.10-20% of asthmatics suffer from severe asthma and don't respond well to standard treatments (eg, β2 agonists, leukotriene antagonists and inhaled steroids). A subset of these patients present with high neutrophil counts in their lungs, and are sensitive to repeated and persistent viral infections that can exacerbate the underlying inflammation and lead to asthma attacks. Inhibitors of SSAO / VAP-1 have been shown to significantly reduce inflammation and airway hyper- reactivity in a mouse model of viral-induced asthma exacerbation. In addition to asthma, SSAO / VAP-1 is associated with other lung inflammatory diseases. Clinically, SSAO / VAP-1 is upregulated in both COPD and idiopathic pulmonary fibrosis patients and preclinically, SSAO / VAP-1 has been shown to be increased in a porcine model of acute respiratory distress syndrome (ARDS). The product of aminoacetone oxidation by SSAO / VAP-1, methyl glyoxal, is significantly elevated in patients admitted to ICU with COVID-19 infections and correlates with mortality, and plasma SSAO / VAP-1 has been observed to be elevated in patients with both mild and severe disease. SSAO / VAP-1 inhibitors have shown beneficial effects in multiple preclinical models of neutrophilic lung inflammation, including lipopolysaccharide-induced lung inflammation and Klebsiella pneumoniae infection where inhibition diminished, but did not abolish neutrophil influx thereby avoiding the immunosuppressive effects associated with corticosteroid use. A SSAO / VAP-1 inhibitor has also been shown to substantially suppress inflammatory cell influx and fibrosisin the airways and improve lung function when dosed therapeutically in a cigarette smoking mouse model of COPD.

[0014] MPO is also associated with a variety of lung inflammatory diseases and conditions, such as ARDS, acute lung injury (ALI), ventilator associated pneumonia and asthma. Increased serum MPO levels are associated with rapid lung function decline and poor cardiovascular outcomes in COPD, and MPO is considered to be a biomarker for disease severity. Late intervention with a MPO inhibitor in a guinea pig model of COPD has been reported to stop progression of emphysema and small airway remodeling. Inhibition of MPO has also been shown to decrease morbidity and oxidative stress in mice with cystic fibrosis-like lung inflammation. MPO has been positively associated with the likelihood of needing to be artificially ventilated in intensive care units when infected with SARS-CoV-2 virus and positively diagnosed with COVID-19.

[0015] As both SSAO / VAP-1 and MPO are implicated in neutrophil-driven inflammation in a variety of lung disease, their inhibition is expected to reduce inflammation which occurs as a consequence of infectious diseases, such as COVID-19 and other viral or bacterial- induced diseases. Concomitant inhibition of both those targets may lead to stronger and / or more extensive effects. A compound that is able to inhibit both those targets concomitantly is highly desirable.

[0016] Serum MPO levels are used as a prognostic marker for the progression of atherosclerosis, and evidence suggests that MPO and MPO-derived oxidants play a part in the development of atherosclerotic plaques. Administration of an MPO inhibitor has been shown to be efficacious in mouse models of atherosclerosis, vasculitis and myocardial infarction. Clinical MPO inhibitor AZD4831 (mitiperstat) has shown benefit in a Phase 2 study in patients suffering from heart failure with a preserved ejection fraction. Circulating SSAO / VAP-1 levels have also been found to correlate with cardiovascular risk factors and early atherosclerotic manifestations. Circulating SSAO / VAP-1 has also been shown to contribute to endothelial progenitor cell dysfunction in patients with vasculitis. SSAO / VAP-1 is expressed in adipocytes where it plays a role in glucose transport independent of the presence of insulin. It has been observed that levels of plasma SSAO / VAP-1 are increased in patients suffering from diabetes and SSAO / VAP-1 has been linked to an increase in cross-linking and advanced glycation end products in diabetic animal models.

[0017] The implications of oxidative stress and reactive oxygen species in fibrosis are well established. The roles of both systemic and localised inflammation and environmental toxins (e.g. cigarette smoke) in pulmonary fibrosis and hepatic fibrosis have been explored.

[0018] In the kidney both SSAO / VAP-1 and MPO are implicated in the pathology of fibrosis and chronic kidney disease. In mice, MPO deficiency ameliorates the progression of chronic kidney disease andSSAO / VAP-1 inhibitors have been shown to significantly reduce fibrosis in two different models of kidney disease.

[0019] Circulating levels of soluble SSAO / VAP-1 are 2-4 fold higher in patients with liver disease (alcoholic liver disease, biliary cirrhosis and hepatocellular carcinoma). Moreover, it has been shown that these levels correct themselves upon transplant of the diseased organ suggesting that the inflamed liver is the source of SSAO / VAP-1 and SSAO / VAP-1 has a significant role in pathophysiology of the diseased organ. Additionally, in multiple murine models of hepatic injury an absence or blockade of SSAO / VAP-1 reduced inflammatory cell recruitment to the liver and attenuated fibrosis. This effect was shown to be dependent on the enzyme activity.

[0020] It has been observed that there is increased hepatic myeloperoxidase activity in obese subjects with NAFLD / NASH. Levels of soluble SSAO / VAP-1 are also elevated in NAFLD / NASH patients and correlate with disease severity, indicating that SSAO / VAP-1 is also a valid biomarker for non-invasive assessment of disease severity. Similarly, a correlation has been shown between NASH and MPO in obese patients, and both MPO- / -mice and inhibitors of MPO activity have been shown to attenuate disease in preclinical studies.

[0021] Inflammation linked to SSAO / VAP-1 enzyme activity is also believed to be an important feature of neuromuscular diseases such as Parkinson’s disease, Alzheimer’s disease and multiple sclerosis, and is a feature of the pathophysiology that occurs after a cerebral occlusion / reperfusion event. Excessive SSAO / VAP-1 activity has been independently implicated in these processes. Pharmacological inhibition of SSAO / VAP-1 activity has been shown to reduce neutrophil infiltration into the brain, and reduce microglial activation in an LPS-induced model of neuroinflammation. It is also thought that endothelial SSAO / VAP-1 plays a role in the cerebral amyloid angiopathy related to Alzheimer’s disease through an effect on cerebrovascular amyloid-beta (Aβ) deposits, leading to the underlying blood-brain barrier dysfunction associated with disease progression. Additionally, SSAO-mediated oxidation of primary amines (such as methylamine to formaldehyde) has been linked to oxidative stress in models of Alzheimer’s disease.

[0022] Complementary to this, MPO has been reported to be upregulated in regions of neurodegeneration in the brains of both Parkinson’s disease and Alzheimer’s disease patients, and the efficacy of MPO inhibitors in experimental models of Parkinson’s disease and multiple systems atrophy (MSA) show the potential of such approaches in the treatment of neurodegenerative diseases.

[0023] Neutrophils have become the focus of new therapeutic approaches for the treatment of muscular dystrophies following studies in mdx mice that suggest that they promote muscle lesion; antibody-mediated depletion of host neutrophils in dystrophic mice resulted in a delayed and significantly reduced amount ofskeletal muscle breakdown. Additionally, it has been shown that neutrophil elastase activity is increased in dystrophic mice and this increase impairs myoblast survival. SSAO / VAP-1 is up-regulated in muscle donor tissue from Duchenne Muscular Dystrophy (DMD) patients, and pharmacological inhibition of SSAO / VAP-1 activity in vivo reduces inflammation in mdx mice, further supporting the theory that neutrophils, recruited at least in part through SSAO-mediated activity, are involved in the progression of muscular dystrophy.

[0024] MPO released by neutrophils in dystrophic muscle has also been indicated to play a role in disease progression, with levels of MPO correlating with the amount of oxidative stress measured in the muscle in a dog model of DMD. Treatment with high dose taurine has been shown to reduce inflammation and reactive oxygen species (ROS), and increase muscle function and protect muscles from necrosis in both mdx mice and in the golden retriever muscular dystrophy model, presumably through “quenching” of the ROS (e.g., HOCl) produced by MPO activity.

[0025] Both SSAO / VAP-1 and MPO are significant targets for therapeutic intervention in their own right. Given the complementary roles that these two enzymes play in the context of neutrophil-driven inflammation, it is proposed that combined targeting (dual therapy) may represent an improved and complementary approach to resolving inflammation driven by an aberrant neutrophilic response. Equally, where both targets are involved in the pathophysiology of disease through oxidative stress and ongoing fibrosis, this dual therapy offers a better chance at improving patient outcomes than mono therapy alone.

[0026] A dual inhibitor refers to a type of drug that can simultaneously target two different molecular targets involved in a particular disease or condition. This approach offers several advantages over combination therapy, which involves using two separate drugs to target different aspects of the disease. Firstly, a dual inhibitor can provide a more targeted and precise treatment approach. By specifically targeting two key molecular targets, it can potentially achieve a greater therapeutic effect and spectrum compared to using two separate drugs that may have overlapping or redundant actions. Secondly, using a single dual inhibitor can simplify the treatment process for patients. Instead of having to take multiple drugs, a patient can benefit from a single medication that targets multiple pathways. This can improve treatment adherence and convenience for the patient. Additionally, a dual inhibitor can also potentially reduce the risk of drug interactions and adverse effects. When two separate drugs are used in combination therapy, there is a higher chance of potential interactions between the drugs, leading to unexpected side effects. With a dual inhibitor, the drug's formulation can be optimized to minimize such interactions. Overall, a dual inhibitor offers the potential for enhanced efficacy, greater proportion of patients responding, simplified treatment, and reduced risks compared to combination therapy.

[0027] Some known MAO inhibitors also inhibit SSAO / VAP-1 (e.g., the MAO-B inhibitor Mofegiline illustrated below). Mofegiline has been reported to inhibit experimental autoimmune encephalomyelitis (US 2006 / 0025438 A1). This inhibitor is a member of the haloallylamine family of MAO inhibitors. Fluoroallylamine inhibitors are also described in US 4,454,158 as MAO inhibitors.

[0028] Other examples structurally related to Mofegiline, such as LJP1586, are described in WO 2007 / 120528:

[0029] Additionally, a series of 2-substituted-3-haloallylamine SSAO / VAP-1 inhibitors has been disclosed in patent application WO 2013 / 163675 including PXS-4728 and PXS-4681:

[0030] In both of these patent applications the enzyme inhibition reported was selective for SSAO / VAP- 1.

[0031] Numerous patent applications have been filed more recently describing additional types of aryloxy and heteroaryloxy 2-substituted-3-fluoroallylamines as SSAO / VAP-1 inhibitors, for example WO 2019 / 241751.

[0032] Inhibition of MPO has been described for a variety of different structural classes.

[0033] Fused 2-thioxo-2,3-dihydropyrimidin-4(1H)-ones such as AZD5904 (imidazole fused - WO2003 / 089430) and AZD3241 / BHV3241 (verdiperstat; pyrrole fused - WO2006 / 0262465) have been extensively studied in the scientific literature in models of MSA, COPD and Parkinson’s disease.

[0034] Further molecules also displaying the pyrrolo-fused core have been described as potent inhibitors of MPO with high selectivity over TPO (WO2016 / 087338). The chloro example shown below has been designated AZD4831 (mitiperstat), and is currently in clinical development.

[0035] Non-fused 2-thioxo-2,3-dihydropyrimidin-4(1H)-ones have been reported in a series of patents describing PF-06282999 (WO2013 / 068875) and derivatives thereof with diminished activity towards the hERG ion channel (WO2016 / 178113).

[0036] There remains a need for compounds that are useful as inhibitors of both SSAO / VAP-1 and MPO. Summary

[0037] The present invention provides haloallylamine fused thiouracil compounds that inhibit the enzyme activity of both SSAO / VAP-1 and MPO. Compounds of the invention were found to show selectivity for the inhibition of SSAO / VAP-1 and MPO compared to inhibition of other amine oxidase family members, such as monoamine oxidases, diamine oxidase, lysyl oxidase (LOX), and lysyl-like amine oxidases LOXL1-4, and also peroxidase family members, such as eosinophil peroxidase, thyroid peroxidase lactoperoxidase, and vascular peroxidase.

[0038] The present invention describes the synthesis and use of compounds which inhibit the amine oxidase activity of SSAO / VAP-1 and the peroxidase activity of MPO, and details the use of such dual- acting inhibitors to treat patients suffering from inflammatory diseases.

[0039] A first aspect of the invention provides for a compound of formula I:or a pharmaceutically acceptable salt thereof; wherein: R1and R2are independently selected from the group consisting of hydrogen and fluorine; provided that R1and R2are not hydrogen at the same time; X is selected from the group consisting of N, CH and C-CH3; Y is selected from the group consisting of hydrogen, fluorine and chlorine; and Z is hydrogen or fluorine.

[0040] A second aspect of the invention provides for a pharmaceutical composition comprising a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient, carrier or diluent.

[0041] A third aspect of the invention provides for a method of inhibiting the amine oxidase activity of SSAO / VAP-1 and the perioxidase activity of MPO in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to the second aspect of the invention.

[0042] A fourth aspect of the invention provides for a method of treating or preventing a disease by inhibiting the activity of the SSAO / VAP-1 protein and the MPO protein, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to the first aspect of the invention or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to the second aspect of the invention.

[0043] A fifth aspect of the invention provides for use of a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating or preventing a disease by inhibiting the activity of the SSAO / VAP-1 protein and the MPO protein.

[0044] A sixth aspect of the invention provides for a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt thereof, for use in treating or preventing a disease by inhibiting the activity of the SSAO / VAP-1 protein and the MPO protein.

[0045] In one embodiment of the methods and uses of the present invention the disease is associated with inflammation or fibrosis.

[0046] In one embodiment of the methods and uses of the present invention the disease is associated with a reduction in kidney function, and the cause of this reduction in kidney function is selected from the group consisting of acute tubular necrosis (ATN), severe or sudden dehydration, toxic kidney injury from poisons or certain medications, the result of a severe bacterial or viral infection, such as COVID-19, urinary tract obstruction and autoimmune kidney diseases, such as acute nephritic syndrome and interstitial nephritis.

[0047] In one embodiment of the methods and uses of the present invention the disease is chronic kidney disease, and the cause of the chronic kidney disease is selected from the group consisting of obesity, type 1 or type 2 diabetes, including diabetic nephropathy, high blood pressure, glomerulonephritis, interstitial nephritis, polycystic kidney disease, prolonged obstruction of the urinary tract from conditions such as cancer, heart failure, heart failure with preserved ejection fraction, heart failure with reduced ejection fraction, vasculitis (including anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis), enlarged prostate or kidney stones, vesicoureteral reflux and pyelonephritis.

[0048] In one embodiment of the methods and uses of the present invention the disease is a respiratory disease selected from the group consisting of asthma, neutrophilic asthma, chronic obstructive pulmonary disease, lung fibrosis, including idiopathic pulmonary fibrosis, bronchiectasis, interstitial lung disease or resulting from severe bacterial or viral infection, mesothelioma, asbestosis, silicosis, effects of severe bacterial or viral infection in the lung such as acute respiratory distress syndrome or acute lung injury as a result of a cytokine storm, ventilator associated pneumonia, and pulmonary complications arising as a result thereof,

[0049] In one embodiment of the methods and uses of the present invention the disease is a cardiovascular disease selected from the group consisting of hypertension, abnormal heart rhythms, or arrhythmias, atherosclerosis, aorta disease and Marfan syndrome, congenital heart disease, cardiomyopathy, coronary artery disease, deep vein thrombosis and pulmonary embolism, dyslipidemias, heart attack, heart failure, heart failure with preserved ejection fraction, heart failure with reduced ejection fraction, cardiac fibrosis,including viral or bacterial induced inflammation and fibrosis, pericardial disease, peripheral vascular disease, rheumatic heart disease, stroke and vasculitis (including anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis).

[0050] In one embodiment of the methods and uses of the present invention the disease is a liver disease selected from the group consisting of liver fibrosis, including alcoholic liver disease, fatty liver disease, non-alcoholic steatohepatitis (NASH) / metabolic dysfunction-associated steatohepatitis (MASH), acute and chronic hepatitis, biliary cirrhosis, primary sclerosing cholangitis, liver autoimmune diseases, hepatocellular carcinoma and toxic liver injury.

[0051] In one embodiment of the methods and uses of the present invention the disease is an eye disease selected from the group consisting of uveitis, diabetic macular oedema, glomerulosclerosis, diabetic retinopathy, age-related macular degeneration, ocular scarring and conjunctival inflammatory disease.

[0052] In one embodiment of the methods and uses of the present invention the disease is selected from the group consisting of a fibrotic disease and a neurological disease.

[0053] Contemplated herein is combination therapy in which the methods further comprise co administering additional therapeutic agents that are used for the treatment of disorders that are associated with cardiovascular disease, kidney disease, respiratory disease, liver disease, eye disease, inflammation or fibrosis. In one embodiment of the methods of the present inventions a compound of the first aspect of the invention may be administered with a second therapeutic agent. In one embodiment the second therapeutic agent is selected from the group consisting of an anti-hyperglycemic agent, an anti-inflammatory agent, an anti-hypertensive agent, an agent used to lower cholesterol, a diuretic agent, an anti-fibrotic agent, an agent used to manage heart disease, an anti-angiogenic agent, an antiviral agent and an immunosuppressive agent. Definitions

[0054] The following are some definitions that may be helpful in understanding the description of the present invention. These are intended as general definitions and should in no way limit the scope of the present invention to those terms alone, but are put forth for a better understanding of the following description.

[0055] Unless the context requires otherwise or specifically states to the contrary, integers, steps, or elements of the invention recited herein as singular integers, steps or elements clearly encompass both singular and plural forms of the recited integers, steps or elements.

[0056] Throughout this specification, unless the context requires otherwise, the word "comprise", or variations such as "comprises" or "comprising", will be understood to imply the inclusion of a stated stepor element or integer or group of steps or elements or integers, but not the exclusion of any other step or element or integer or group of elements or integers. Thus, in the context of this specification, the term “comprising” means “including principally, but not necessarily solely”.

[0057] Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the invention includes all such variations and modifications. The invention also includes all of the steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations or any two or more of said steps or features.

[0058] The present invention includes within its scope isotopes of different atoms. Any atom not specifically designated as a particular isotope is meant to represent any stable isotope of that atom. Thus, the present disclosure should be understood to include the deuterium isotope of hydrogen.

[0059] All references cited in this application are specifically incorporated by cross-reference in their entirety. Reference to any such documents should not be construed as an admission that the document forms part of the common general knowledge or is prior art.

[0060] In the context of this specification the term “administering” and variations of that term including “administer” and “administration”, includes contacting, applying, delivering or providing a compound or composition of the invention to an organism, or a surface by any appropriate means. In the context of this specification, the term “treatment”, refers to any and all uses which remedy a disease state or symptoms, prevent the establishment of disease, or otherwise prevent, hinder, retard, or reverse the progression of disease or other undesirable symptoms in any way whatsoever.

[0061] In the context of this specification the term “effective amount” includes within its meaning a sufficient but non-toxic amount of a compound or composition of the invention to provide a desired effect. Thus, the term “therapeutically effective amount” includes within its meaning a sufficient but non-toxic amount of a compound or composition of the invention to provide the desired therapeutic effect. The exact amount required will vary from subject to subject depending on factors such as the species being treated, the sex, age and general condition of the subject, the severity of the condition being treated, the particular agent being administered, the mode of administration, and so forth. Thus, it is not possible to specify an exact “effective amount”. However, for any given case, an appropriate “effective amount” may be determined by one of ordinary skill in the art using only routine experimentation.Brief Description of the Figures

[0062] Figure 1 depicts the anti-inflammatory effects of Compound 1 in BALB / cJUnib mice in a model of acute lung inflammation.

[0063] Figure 2 depicts the anti-inflammatory effects of Compound 1 in C57BL / 6 mice in a model of peritoneal inflammation.

[0064] Figure 3 depicts the ability of Compound 1 to inhibit MPO in the peritoneum in C57BL / 6 mice in a model of peritoneal inflammation.

[0065] Figure 4 depicts the anti-inflammatory effects of Compound 1 in BALB / c mice in the carrageenan air pouch model.

[0066] Figure 5 depicts the ability of Compound 1 to inhibit SSAO / VAP-1 in the adipose tissue in BALB / c mice in the carrageenan air pouch model.

[0067] Figure 6 depicts the anti-fibrotic effects of Compound 1 in Wistar rats in a model of cardiac ischemia- reperfusion injury.

[0068] Figure 7 depicts the anti-fibrotic effects of Compound 1 in C57 / Bl6J mice in the unilateral ureteral obstruction model. Detailed Description

[0069] The present invention relates to substituted haloallylamine derivatives which may inhibit both SSAO / VAP-1 and MPO. In particular, the present invention relates to substituted fluoroallylamine derivatives with an attached thiouracil moiety.

[0070] In particular, the present invention relates to compounds of formula I:or a pharmaceutically acceptable salt thereof; wherein:R1and R2are independently selected from the group consisting of hydrogen and fluorine; provided that R1and R2are not hydrogen at the same time; X is selected from the group consisting of N, CH and C-CH3; Y is selected from the group consisting of hydrogen, fluorine and chlorine; and Z is hydrogen or fluorine.

[0071] In one embodiment of the compounds of the present invention X is selected from the group consisting of N, CH and C-CH3. In another embodiment of the compounds of the present invention X is N or CH. In a further embodiment of compounds of the present invention X is N or C-CH3. In another embodiment of compounds of the present invention X is CH or C-CH3. In a further embodiment of the compounds of the present invention X is N. In another embodiment of the compounds of the present invention X is CH. In a further embodiment of the compounds of the present invention X is C-CH3.

[0072] In one embodiment of the compounds of the present invention R1and R2are independently selected from the group consisting of hydrogen and fluorine, provided that R1and R2are not hydrogen at the same time. In another embodiment of the compounds of the present invention R1is hydrogen and R2is fluorine. In a further embodiment of the compounds of the present invention R1is fluorine and R2is hydrogen. In another embodiment of the compounds of the present invention R1and R2are both fluorine.

[0073] In one embodiment of the compounds of the present invention Y is selected from the group consisting of hydrogen, fluorine and chlorine. In another embodiment of the compounds of the present invention Y is hydrogen or fluorine. In a further embodiment of compounds of the present invention Y is hydrogen or chlorine. In another embodiment of compounds of the present invention Y is fluorine or chlorine. In a further embodiment of the compounds of the present invention Y is hydrogen. In another embodiment of the compounds of the present invention Y is fluorine. In a further embodiment of the compounds of the present invention Y is chlorine.

[0074] In one embodiment of the compounds of the present invention Z is hydrogen or fluorine. In another embodiment of the compounds of the present invention Z is hydrogen. In a further embodiment of the compounds of the present invention Z is fluorine.

[0075] In one embodiment of the compounds of the present invention X is N, R1and R2are independently selected from the group consisting of hydrogen and fluorine, provided that R1and R2are not hydrogen at the same time, Y is hydrogen and Z is hydrogen. In another embodiment of the compounds of the present invention X is N, R1is hydrogen, R2is fluorine, Y is hydrogen and Z is hydrogen. In a further embodimentof the compounds of the present invention X is N, R1is fluorine, R2is hydrogen, Y is hydrogen and Z is hydrogen.

[0076] In one embodiment of the compounds of the present invention X is C-CH3, R1is hydrogen, R2is fluorine, Y is selected from the group consisting of hydrogen, fluorine and chlorine and Z is hydrogen or fluorine. In another embodiment of the compounds of the present invention X is C-CH3, R1is hydrogen, R2is fluorine, Y is hydrogen and Z is fluorine. In a further embodiment of the compounds of the present invention X is C-CH3, R1is hydrogen, R2is fluorine, Y is fluorine and Z is hydrogen. In another embodiment of the compounds of the present invention X is C-CH3, R1is hydrogen, R2is fluorine, Y is chlorine and Z is hydrogen.

[0077] In one embodiment of the compounds of the present invention X is CH, R1is hydrogen, R2is fluorine, Y is hydrogen or fluorine and Z is hydrogen or fluorine. In another embodiment of the compounds of the present invention X is CH, R1is hydrogen, R2is fluorine, Y is hydrogen and Z is fluorine. In a further embodiment of the compounds of the present invention X is CH, R1is hydrogen, R2is fluorine, Y is fluorine and Z is hydrogen. In another embodiment of the compounds of the present invention X is CH, R1is hydrogen, R2is fluorine, Y is hydrogen and Z is hydrogen.

[0078] In one embodiment of the present invention there is provided a compound of Formula Ia:or a pharmaceutically acceptable salt thereof; wherein: X is selected from the group consisting of N, CH and C-CH3; Y is selected from the group consisting of hydrogen, fluorine and chlorine; and Z is hydrogen or fluorine.

[0079] In the context of the present disclosure, any one or more aspect(s) or embodiment(s) may be combined with any other aspect(s) or embodiment(s).

[0080] Exemplary compounds according to the present invention include the compounds set forth in Table 1:Table 1 Compound - -- -Preparation of Compounds of Formula I

[0081] Compounds of Formula I described herein are synthesized using standard synthetic chemistry techniques using methods known in the art. The methods described are provided for illustrative purposes only and do not limit the scope of the claims provided herein.

[0082] Standard synthetic chemistry techniques are described in, but not limited to, Advanced Organic Chemistry, 8th Edition by March, John Wiley and Sons Inc. Standard procedures for the use of protecting groups for the temporary protection of functional groups such as alcohols, amines and carboxylic acids are described in for example Protecting Groups in Organic Synthesis, 5th Edition, John Wiley and Sons Inc.

[0083] Alternative reaction conditions for the chemical transformations described herein may be employed such as variation in solvent, reaction temperature, reaction time as well as different chemical reagents. Unless stated otherwise, starting materials for chemical synthesis and biological applications are available from commercial sources.

[0084] In general scheme 1, P1is a functional group used to protect nitrogen or sulphur functionality. In some embodiments, P1is a benzyl or 4-methoxybenzyl (PMB) group. In general schemes 1 and 2, P2and / or P3is a functional group used to protect a nitrogen functionality. In some embodiments, P2and / or P3are alkoxycarbonyl groups such as the tert-butyloxycarbonyl (BOC), the 9-fluorenylmethyloxycarbonyl (FMOC), and the benzyloxycarbonyl (CBZ) groups. In some embodiments, P2and P3form a cyclic nitrogen protecting group such as N-phthalimide and N-2,5-dimethylpyrrole. In some embodiments, compounds indicated by Formula I (X = N) are prepared as shown by Scheme 1. In some embodiments, compounds of general structure 1-4 can be obtained through a process of reductive amination. In some embodiments, 3-formylated phenols of structure 1-1 are treated with 4-amino-1H-imidazole-carboxamide (1-2) in a suitable solvent such as MeOH at a temperature of 0 – 30 °C, for 8 - 20 h to provide intermediate of generalstructure 1-3. In an alternative embodiment, the solvent is EtOH. In some embodiments, subsequent reduction of the imine functional group of compounds of general structure 1-3, to provide structure 1-4 is achieved by treatment with Pd / C in the presence of an atmosphere of H2, in a suitable solvent such as MeOH or THF (or combination thereof). In some embodiments, intermediate of general structure 1-3 is treated with a reducing agent such as NaBH3CN to provide compounds of structure 1-4. In yet another embodiment, the reducing agent is NaBH4. In an alternative embodiment, compounds of structure 1-1 can be converted to compounds of structure 1-4 in one “pot” without intermediate work-up or isolation of compounds of structure 1-3. In some embodiments, compounds of structure 1-4 are treated with benzoyl isothiocyanate in a suitable solvent such as CH2Cl2 or MeOH (or combination thereof) to provide compounds of general structure 1-5. In some embodiments, subsequent treatment of 1-5 with a suitable base, for example K2CO3 or Cs2CO3, in a solvent such as water or MeOH at 20 – 110 °C, provides thiouracil compounds of general structure 1-6. In another embodiment the base is NaOH or Na2CO3. In a further embodiment, compounds of general structure 1-4 can be converted to compounds of general structure 1-6 without isolation of the intermediate of general structure 1-5. In some embodiments, compounds of structure 1-6 can be suitably protected to provide compounds of general structure 1-7. In some embodiments, the protecting group (P1) is a 4-methoxybenzyl group. In some embodiments, compounds of structure 1-6 are treated with 4- methoxybenzyl chloride in a suitable solvent such as dimethylformamide (DMF) to provide compounds of structure 1-7. In some embodiments, compounds of structure 1-7 are treated with allylbromides of structure 1-8 in the presence of a base, for example Cs2CO3 in a solvent such as DMF at 20 – 40 °C to provide compounds of structure 1-9. In an alternative embodiment, the base is K2CO3. In yet another embodiment, the base is NaOtBu. There are many, well established, chemical procedures for the de-protection of compounds of the general structure 1-9 to compounds described by Formula I. For example, if P1is a PMB protecting group and P2and / or P3is a BOC protecting group, compounds described by structure 1-9 can be treated with an acidic substance such as dry hydrogen chloride in a suitable solvent such as diethyl ether or ethyl acetate or 1,4-dioxane to furnish the compounds described by Formula I as the hydrochloride salt. In another embodiment, the acidic reagent is trifluoroacetic acid (TFA). In yet another embodiment, compounds of structure 1-9 are treated with a combination of TFA and dry HCl in a solvent such as ethyl acetate or 1,4-dioxane. In general, the free amino compounds are converted to acid addition salts for ease of handling and for improved chemical stability. Examples of acid addition salts include, but are not limited to, hydrochloride, hydrobromide, 2,2,2-trifluoroacetate and methanesulfonate salts.Scheme 1

[0085] In some embodiments, compounds indicated by Formula I (X = C or C-CH3) are prepared as shown by Scheme 2. In some embodiments, 3-formylated phenols of structure 2-1 are treated with allylbromides of structure 2-2 in the presence of a base, for example Cs2CO3, in a solvent such as DMF at 20 – 40 °C to provide compounds of structure 2-3. In an alternative embodiment, the base is K2CO3. In yet another embodiment, the base is NaOtBu. In some embodiments, compounds of general structure 2-5 can be obtained through a process of reductive amination. In some embodiments, compounds of structure 2-3 are treated with ethyl 4-aminopyrrole-carboxylates of general structure 2-4, in the presence of a suitable reducing agent such as NaBH4or NaBH3CN, in a suitable solvent such as MeOH at a temperature of 0 –45 °C, for 2 - 10 h to provide compounds of structure 2-5. In an alternative embodiment, the solvent is EtOH. In some embodiments, compounds of structure 2-5 are treated with benzoyl isothiocyanate in a suitable solvent such as CH2Cl2 or MeOH (or combination thereof) to provide compounds of general structure 2-6. In some embodiments, subsequent treatment of 2-6 with a suitable base, for example K2CO3 or Cs2CO3, in a solvent such as water or MeOH at 20 – 110 °C, provides thiouracil compounds of general structure 2-7. In another embodiment the base is NaOH or Na2CO3. In a further embodiment, compounds of general structure 2-5 can be converted to compounds of general structure 2-7 without isolation of the intermediate of general structure 2-6. There are many, well established, chemical procedures for the de- protection of compounds of the general structure 2-7 to compounds described by Formula I. For example, if P2and / or P3is a BOC protecting group, compounds of general structure 2-7 can be treated with an acidic substance such as dry hydrogen chloride in a suitable solvent such as diethyl ether or ethyl acetate or 1,4- dioxane to furnish the compounds described by Formula I as the hydrochloride salt. In another embodiment, the acidic reagent is trifluoroacetic acid (TFA). In general, the free amino compounds are converted to acid addition salts for ease of handling and for improved chemical stability. Examples of acid addition salts include, but are not limited to, hydrochloride, hydrobromide, 2,2,2-trifluoroacetate and methanesulfonate salts.Scheme 2 Therapeutic uses

[0086] The present invention provides methods for the use of compounds described by Formula I to inhibit membrane-bound SSAO / VAP-1 and soluble SSAO / VAP-1. In addition, the present invention provides methods for the use of compounds described by Formula I to inhibit MPO. The relative inhibitory potencies of the compounds can be determined by the amount needed to inhibit the enzyme activity of SSAO / VAP-1 and MPO in a variety of ways, e.g., in an in vitro assay with recombinant human protein or with recombinant non-human enzyme, in cellular assays expressing normal rodent enzyme, in cellular assays which have been transfected with human protein, in in vivo tests in rodent and other mammalian species, and the like.

[0087] The present invention also discloses methods to use the compounds described by Formula I to inhibit SSAO / VAP-1 and MPO in patients suffering from an inflammatory disease, and methods to treat inflammatory diseases.

[0088] Thus, in one aspect, the present invention is directed to a method of inhibiting the enzyme activity of SSAO / VAP-1 and MPO in a subject in need thereof, comprising administering to the subject an effective amount of a compound of Formula I, or a pharmaceutically acceptable salt, or solvate thereof, or a pharmaceutical composition thereof.

[0089] In another aspect, the present invention is directed to a method of treating or preventing a disease by inhibiting the activity of the SSAO / VAP-1 protein and the MPO protein, comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition thereof. In one embodiment the present invention is directed to a method of treating a disease by inhibiting the activity of the SSAO / VAP-1 protein and the MPO protein. In another embodiment the present invention is directed to a method of preventing a disease by inhibiting the activity of the SSAO / VAP-1 protein and the MPO protein.

[0090] In still another aspect, the present invention is directed to methods of treating a disease modulated by SSAO / VAP-1 and MPO, said methods comprising administering to a subject in need thereof a therapeutically effective amount of a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof, or a pharmaceutical composition thereof.

[0091] A further aspect of the present invention provides for use of a compound of Formula I, or a pharmaceutically acceptable salt or solvate thereof, for the manufacture of a medicament for treating or preventing a disease by inhibiting the activity of the SSAO / VAP-1 protein and the MPO protein.

[0092] The above-described methods and uses are applicable wherein the disease is inflammation. As employed herein, “inflammation” or “inflammatory disorder” embraces a wide variety of indications, including arthritis (including juvenile rheumatoid arthritis), Crohn’s disease, ulcerative colitis, inflammatory bowel diseases (e.g., irritable bowel disease), psoriasis, asthma, pulmonary inflammation, COPD, bronchiectasis, skin inflammation, ocular disease, contact dermatitis, liver inflammation, liver autoimmune diseases, autoimmune hepatitis, primary biliary cirrhosis, sclerosing cholangitis, autoimmune cholangitis, alcoholic liver disease, non-alcoholic steatohepatitis (NASH) / metabolic dysfunction- associated steatohepatitis (MASH), arteriosclerosis, vasculitis, chronic heart failure, congestive heart failure, ischemic diseases, stroke and complications thereof, inflammation arriving from bacterial or viral infection such as those experienced in sepsis, acute lung injury, acute respiratory distress syndrome,COVID-19 and cystic fibrosis, myocardial infarction and complications thereof, inflammatory cell destruction following stroke, synovitis and systemic inflammatory response syndrome.

[0093] The dual benefits of SSAO / VAP-1 and MPO inhibition include the potential to treat inflammatory disorders related to oxidative stress.

[0094] Diseases associated with the term “oxidative stress” cover disorders where an increase in reactive oxygen species has been linked to the disease progression or to the symptoms of the disease and include a number of different diseases including cancer, muscular dystrophy, Alzheimer’s disease, Parkinson’s disease, diabetes, cardiovascular conditions such as high blood pressure, arteriosclerosis and stroke, inflammatory disorders, chronic fatigue syndrome, Long COVID, fragile X syndrome, depression, attention deficit hyperactivity disorder, autism, Asperger syndrome, Huntington’s disease, amyotrophic lateral sclerosis, multiple sclerosis, asthma and male infertility.

[0095] In one embodiment of the methods and uses of the present invention the disease is associated with a reduction in kidney function, and the cause of this reduction in kidney function is selected from the group consisting of acute tubular necrosis (ATN), severe or sudden dehydration, toxic kidney injury from poisons or certain medications, the result of a severe bacterial or viral infection, such as COVID-19, urinary tract obstruction and autoimmune kidney diseases, such as acute nephritic syndrome and interstitial nephritis.

[0096] In one embodiment of the methods and uses of the present invention the inflammation and oxidative stress are associated with kidney disease, both acute kidney failure and chronic kidney disease.

[0097] Inflammatory disorders and diseases linked to acute kidney failure include, but are not restricted to, acute tubular necrosis (ATN), severe or sudden dehydration, toxic kidney injury from poisons or certain medications, COVID-19, urinary tract obstruction and autoimmune kidney diseases, such as acute nephritic syndrome and interstitial nephritis.

[0098] Inflammatory disorders and disease linked to chronic kidney disease include, but are not restricted to, obesity, type 1 or type 2 diabetes, including diabetic nephropathy, high blood pressure, glomerulonephritis, interstitial nephritis, polycystic kidney disease, prolonged obstruction of the urinary tract from conditions such as cancer, heart failure, heart failure with preserved ejection fraction, heart failure with reduced ejection fraction, vasculitis (including anti-neutrophil cytoplasmic antibody (ANCA)- Associated vasculitis), enlarged prostate or kidney stones, vesicoureteral reflux and pyelonephritis.

[0099] In one embodiment of the method and uses of the present invention the disease is a respiratory disease selected from the group consisting of asthma, neutrophilic asthma, chronic obstructive pulmonary disease, lung fibrosis, including idiopathic pulmonary fibrosis, bronchiectasis, interstitial lung disease orresulting from severe bacterial or viral infection, mesothelioma, asbestosis, silicosis, effects of severe bacterial or viral infection in the lung such as acute respiratory distress syndrome or acute lung injury as a result of a cytokine storm, ventilator associated pneumonia, and pulmonary complications arising as a result thereof.

[0100] In one embodiment of the methods and uses of the present invention the inflammation and oxidative stress are associated with cardiovascular disease.

[0101] As employed herein, “cardiovascular disease” embraces a wide variety of indications, including hypertension, abnormal heart rhythms, or arrhythmias, atherosclerosis, aorta disease and Marfan syndrome, congenital heart disease, cardiomyopathy, coronary artery disease, deep vein thrombosis and pulmonary embolism, dyslipidemias, heart attack, heart failure, heart failure with preserved ejection fraction, heart failure with reduced ejection fraction, cardiac fibrosis, including viral or bacterial induced inflammation and fibrosis, pericardial disease, peripheral vascular disease, rheumatic heart disease, stroke and vasculitis (including anti-neutrophil cytoplasmic antibody (ANCA)-Associated vasculitis).

[0102] In one embodiment of the methods and uses of the present invention the disease is an inflammatory condition selected from a neurological disease, a metabolic disease and fibrosis.

[0103] The above-described methods and uses are also applicable wherein the disease is a neurological disease or disorder. As employed herein, “neurological disease or disorder” covers both neuroinflammatory and neurodegenerative diseases and embraces a variety of indications, including stroke, Alzheimer’s disease, familial Alzheimer's disease, Parkinson’s disease, senile dementia, vascular dementia, Huntington’s disease, amyotrophic lateral sclerosis, multiple sclerosis, Down's syndrome and homozygotes for the apolipoprotein E4 allele.

[0104] The above-described methods and uses are also applicable wherein the disease is a metabolic disorder or disease. As employed here “metabolic disorder or disease” embraces a wide variety of indications, including such diseases as obesity, Type 1 and 2 diabetes, hypertension, atherosclerosis, high blood pressure, nonalcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH) / metabolic dysfunction-associated steatohepatitis (MASH), chronic kidney disease, dyslipidemias, heart disease, metabolic syndrome, insulin resistance, diabetic nephropathy, glomerulosclerosis, diabetic retinopathy, choroidal neovascularization and cardiovascular complications arising as a result thereof.

[0105] In accordance with yet another aspect of the present invention, there are provided methods for treating patients suffering from Type 1 and 2 diabetes, and diabetes-related diseases with a dual SSAO / VAP-1 and MPO inhibitor. Diabetes-related diseases contemplated for treatment herein includediabetic nephropathy, diabetic macular oedema, glomerulosclerosis, diabetic retinopathy, choroidal neovascularization and cardiovascular complications arising as a result thereof.

[0106] The above-described methods and uses are also applicable wherein the disease is fibrosis. As employed here “fibrosis” includes such diseases as cystic fibrosis, idiopathic pulmonary fibrosis, kidney fibrosis, chronic obstructive pulmonary disease, pleural fibrosis and radiation-induced pulmonary fibrosis, non-diabetic renal fibrosis, pancreatic fibrosis, scleroderma, connective tissue diseases, scarring, skin fibrosis, fibrosis associated with organ transplant, stenosis, liver fibrosis, including alcoholic liver disease, fatty liver disease, non-alcoholic steatohepatitis (NASH) / metabolic dysfunction-associated steatohepatitis (MASH), acute and chronic hepatitis, biliary cirrhosis, primary sclerosing cholangitis, hepatocellular carcinoma and toxic liver injury, and other diseases where excessive fibrosis contributes to disease pathology.

[0107] In one embodiment of the method and uses of the present invention the disease is a liver disease selected from the group consisting of liver fibrosis, including alcoholic liver disease, fatty liver disease, non-alcoholic steatohepatitis (NASH) / metabolic dysfunction-associated steatohepatitis (MASH), acute and chronic hepatitis, biliary cirrhosis, primary sclerosing cholangitis, liver autoimmune diseases, hepatocellular carcinoma and toxic liver injury.

[0108] In one embodiment of the method and uses of the present invention the disease is an eye disease selected from the group consisting of uveitis, diabetic macular oedema, glomerulosclerosis, diabetic retinopathy, age-related macular degeneration, ocular scarring and conjunctival inflammatory disease.

[0109] In one embodiment of the method and uses of the present invention the disease is selected from the group consisting of inflammatory bowel disease, a fibrotic disease and a neurological disease. Pharmaceutical and / or Therapeutic Formulations

[0110] In another embodiment of the present invention, there are provided compositions comprising a compound having Formula I and at least one pharmaceutically acceptable excipient, carrier or diluent thereof. The compound(s) of Formula I may also be present as suitable salts, including pharmaceutically acceptable salts.

[0111] The phrase “pharmaceutically acceptable carrier” refers to any carrier known to those skilled in the art to be suitable for the particular mode of administration. In addition, the compounds may be formulated as the sole pharmaceutically active ingredient in the composition or may be combined with other active ingredients.

[0112] The phrase “pharmaceutically acceptable salt” refers to any salt preparation that is appropriate for use in a pharmaceutical application. By pharmaceutically acceptable salt it is meant those salts which, within the scope of sound medical judgement, are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit / risk ratio.

[0113] Pharmaceutically acceptable salts are well known in the art and include acid addition and base salts. Hemisalts of acids and bases may also be formed. Pharmaceutically acceptable salts include amine salts of mineral acids (e.g., hydrochlorides, hydrobromides, sulfates, and the like); and amine salts of organic acids (e.g., formates, acetates, lactates, malates, tartrates, citrates, ascorbates, succinates, maleates, butyrates, valerates, fumarates, and the like).

[0114] As compounds of Formula I have a basic site, suitable pharmaceutically acceptable salts may be acid addition salts. For example, suitable pharmaceutically acceptable salts of such compounds may be prepared by mixing a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, methanesulfonic acid, succinic acid, fumaric acid, maleic acid, benzoic acid, phosphoric acid, acetic acid, oxalic acid, carbonic acid, tartaric acid, or citric acid with the compounds of the invention.

[0115] S. M. Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66:1-19. The salts can be prepared in situ during the final isolation and purification of the compounds of the invention, or separately by reacting the free base function with a suitable organic acid. Representative acid addition salts include acetate, adipate, alginate, ascorbate, asparate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, digluconate, cyclopentanepropionate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate salts, and the like. Suitable base salts are formed from bases that form non-toxic salts. Examples include the aluminium, arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine and zinc salts. Representative alkali or alkaline earth metal salts include sodium, lithium potassium, calcium, magnesium, and the like, as well as non-toxic ammonium, quaternary ammonium, and amine cations, including, but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, triethanolamine and the like.

[0116] Pharmaceutically acceptable salts of compounds of Formula I may be prepared by methods known to those skilled in the art, including for example: i. by reacting the compound of formula I with the desired acid or base; ii.by removing an acid- or base-labile protecting group from a suitable precursor of the compound of formula I or by ring-opening a suitable cyclic precursor, for example, a lactone or lactam, using the desired acid or base; or iii. by converting one salt of the compound of formula I to another by reaction with an appropriate acid or base or by means of a suitable ion exchange column.

[0117] The above reactions (i)-(iii) are typically carried out in solution. The resulting salt may precipitate out and be collected by filtration or may be recovered by evaporation of the solvent. The degree of ionisation in the resulting salt may vary from completely ionised to almost non-ionised.

[0118] Thus, for instance, suitable pharmaceutically acceptable salts of compounds according to the present invention may be prepared by mixing a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, methanesulfonic acid, succinic acid, fumaric acid, maleic acid, benzoic acid, phosphoric acid, acetic acid, oxalic acid, carbonic acid, tartaric acid, or citric acid with the compounds of the invention. Suitable pharmaceutically acceptable salts of the compounds of the present invention therefore include acid addition salts.

[0119] The compounds of the invention may exist in both unsolvated and solvated forms. The term ‘solvate’ is used herein to describe a molecular complex comprising the compound of the invention and a stoichiometric amount of one or more pharmaceutically acceptable solvent molecules, for example, ethanol. The term ‘hydrate’ is employed when the solvent is water.

[0120] In one embodiment the compounds of Formula I may be administered in the form of a “prodrug”. The phrase “prodrug” refers to a compound that, upon in vivo administration, is metabolized by one or more steps or processes or otherwise converted to the biologically, pharmaceutically or therapeutically active form of the compound. Prodrugs can be prepared by modifying functional groups present in the compound in such a way that the modifications are cleaved, either in routine manipulation or in vivo, to a compound described herein. For example, prodrugs include compounds of the present invention wherein a hydroxy, amino, or sulfhydryl group is bonded to any group that, when administered to a mammalian subject, can be cleaved to form a free hydroxyl, free amino, or free sulfhydryl group, respectively. Representative prodrugs include, for example, amides, esters, enol ethers, enol esters, acetates, formates, benzoate derivatives, and the like of alcohol and amine functional groups in the compounds of the present invention. The prodrug form can be selected from such functional groups as -C(O)alkyl, -C(O)cycloalkyl, -C(O)aryl, -C(O)- arylalkyl, C(O)heteroaryl, -C(O)-heteroarylalkyl, or the like. By virtue of knowledge of pharmacodynamicprocesses and drug metabolism in vivo, those of skill in this art, once a pharmaceutically active compound is known, can design prodrugs of the compound (see, e.g., Nogrady (1985) Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392).

[0121] Compositions herein comprise one or more compounds provided herein. The compounds are, in one embodiment, formulated into suitable pharmaceutical preparations such as solutions, suspensions, tablets, creams, gels, dispersible tablets, pills, capsules, powders, sustained release formulations or elixirs, for oral administration or in sterile solutions or suspensions for parenteral administration, as well as transdermal patch preparation and dry powder inhalers. In one embodiment, the compounds described above are formulated into pharmaceutical compositions using techniques and procedures well known in the art (see, e.g., Ansel Introduction to Pharmaceutical Dosage Forms, Fourth Edition 1985, 126).

[0122] In the compositions, effective concentrations of one or more compounds or pharmaceutically acceptable derivatives thereof is (are) mixed with a suitable pharmaceutical carrier. The compounds may be derivatized as the corresponding salts, esters, enol ethers or esters, acetals, ketals, orthoesters, hemiacetals, hemiketals, acids, bases, solvates, hydrates or prodrugs prior to formulation, as described above. The concentrations of the compounds in the compositions are effective for delivery of an amount, upon administration, that treats, prevents, or ameliorates one or more of the symptoms of diseases or disorders to be treated.

[0123] In one embodiment, the compositions are formulated for single dosage administration. To formulate a composition, the weight fraction of compound is dissolved, suspended, dispersed or otherwise mixed in a selected carrier at an effective concentration such that the treated condition is relieved, prevented, or one or more symptoms are ameliorated.

[0124] The active compound is included in the pharmaceutically acceptable carrier in an amount sufficient to exert a therapeutically useful effect in the absence of undesirable side effects on the patient treated. The therapeutically effective concentration may be determined empirically by testing the compounds in in vitro and in vivo systems described herein and in PCT publication WO 04 / 018997, and then extrapolated from there for dosages for humans.

[0125] The concentration of active compound in the pharmaceutical composition will depend on absorption, distribution, inactivation and excretion rates of the active compound, the physicochemical characteristics of the compound, the dosage schedule, and amount administered as well as other factors known to those of skill in the art.

[0126] In one embodiment, a therapeutically effective dosage should produce a serum concentration of active ingredient of from about 0.1 ng / mL to about 50 - 100 µg / mL. The pharmaceutical compositions, inanother embodiment, should provide a dosage of from about 0.001 mg to about 2000 mg of compound per kilogram of body weight per day. Pharmaceutical dosage unit forms are prepared to provide from about 0.01 mg, 0.1 mg or 1 mg to about 500 mg, 1000 mg or 2000 mg, and in one embodiment from about 10 mg to about 500 mg of the active ingredient or a combination of essential ingredients per dosage unit form.

[0127] Dosing may occur at intervals of minutes, hours, days, weeks, months or years or continuously over any one of these periods. Suitable dosages lie within the range of about 0.1 ng per kg of body weight to 1 g per kg of body weight per dosage. The dosage is preferably in the range of 1 µg to 1 g per kg of body weight per dosage, such as is in the range of 1 mg to 1 g per kg of body weight per dosage. Suitably, the dosage is in the range of 1 µg to 500 mg per kg of body weight per dosage, such as 1 µg to 200 mg per kg of body weight per dosage, or 1 µg to 100 mg per kg of body weight per dosage. Other suitable dosages may be in the range of 1 mg to 250 mg per kg of body weight, including 1 mg to 10, 20, 50 or 100 mg per kg of body weight per dosage or 10 µg to 100 mg per kg of body weight per dosage.

[0128] Suitable dosage amounts and dosing regimens can be determined by the attending physician and may depend on the particular condition being treated, the severity of the condition, as well as the general health, age and weight of the subject.

[0129] In instances in which the compounds exhibit insufficient solubility, methods for solubilizing compounds may be used. Such methods are known to those of skill in this art, and include, but are not limited to, using cosolvents, such as dimethyl sulfoxide (DMSO), using surfactants, such as TWEEN®, dissolution in aqueous sodium bicarbonate, formulating the compounds of interest as nanoparticles, and the like. Derivatives of the compounds, such as prodrugs of the compounds may also be used in formulating effective pharmaceutical compositions.

[0130] Upon mixing or addition of the compound(s), the resulting mixture may be a solution, suspension, emulsion or the like. The form of the resulting mixture depends upon a number of factors, including the intended mode of administration and the solubility of the compound in the selected carrier or vehicle. The effective concentration is sufficient for ameliorating the symptoms of the disease, disorder or condition treated and may be empirically determined.

[0131] The pharmaceutical compositions are provided for administration to humans and animals in unit dosage forms, such as tablets, capsules, pills, powders, granules, sterile parenteral solutions or suspensions, and oral solutions or suspensions, and oil-water emulsions containing suitable quantities of the compounds or pharmaceutically acceptable derivatives thereof. The pharmaceutically active compounds and derivatives thereof are, in one embodiment, formulated and administered in unit-dosage forms or multiple- dosage forms. The active ingredient may be administered at once, or may be divided into a number of smaller doses to be administered at intervals of time. Unit-dose forms as used herein refers to physicallydiscrete units suitable for human and animal subjects and packaged individually as is known in the art. Each unit-dose contains a predetermined quantity of the therapeutically active compound sufficient to produce the desired therapeutic effect, in association with the required pharmaceutical carrier, vehicle or diluent. Examples of unit-dose forms include ampules and syringes and individually packaged tablets or capsules. Unit-dose forms may be administered in fractions or multiples thereof. A multiple-dose form is a plurality of identical unit-dosage forms packaged in a single container to be administered in segregated unit-dose form. Examples of multiple-dose forms include vials, bottles of tablets or capsules or bottles of pints or gallons. Hence, multiple dose form is a multiple of unit-doses which are not segregated in packaging.

[0132] Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, Pa., 15th Edition, 1975.

[0133] Dosage forms or compositions containing active ingredient in the range of 0.005% to 100% (wt%) with the balance made up from non-toxic carrier may be prepared. Methods for preparation of these compositions are known to those skilled in the art. The contemplated compositions may contain 0.001%- 100% (wt%) active ingredient, in one embodiment 0.1-95% (wt%), in another embodiment 75-85% (wt%). Modes of Administration

[0134] Convenient modes of administration include injection (subcutaneous, intravenous, etc.), oral administration, inhalation, transdermal application, topical creams or gels or powders, vaginal or rectal administration. Depending on the route of administration, the formulation and / or compound may be coated with a material to protect the compound from the action of enzymes, acids and other natural conditions which may inactivate the therapeutic activity of the compound. The compound may also be administered parenterally or intraperitoneally. Compositions for oral administration

[0135] Oral pharmaceutical dosage forms are either solid, gel or liquid. The solid dosage forms are tablets, capsules, granules, and bulk powders. Types of oral tablets include compressed, chewable lozenges and tablets which may be enteric-coated, sugar-coated or film-coated. Capsules may be hard or soft gelatin capsules, while granules and powders may be provided in non-effervescent or effervescent form with the combination of other ingredients known to those skilled in the art. Solid compositions for oral administration

[0136] In certain embodiments, the formulations are solid dosage forms, in one embodiment, capsules or tablets. The tablets, pills, capsules, troches and the like can contain one or more of the followingingredients, or compounds of a similar nature: a binder; a lubricant; a diluent; a glidant; a disintegrating agent; a coloring agent; a sweetening agent; a flavoring agent; a wetting agent; an emetic coating; and a film coating. Examples of binders include microcrystalline cellulose, gum tragacanth, glucose solution, acacia mucilage, gelatin solution, molasses, polvinylpyrrolidine, povidone, crospovidones, sucrose and starch paste. Lubricants include talc, starch, magnesium or calcium stearate, lycopodium and stearic acid. Diluents include, for example, lactose, sucrose, starch, kaolin, salt, mannitol and dicalcium phosphate. Glidants include, but are not limited to, colloidal silicon dioxide. Disintegrating agents include crosscarmellose sodium, sodium starch glycolate, alginic acid, corn starch, potato starch, bentonite, methylcellulose, agar and carboxymethylcellulose. Coloring agents include, for example, any of the approved certified water soluble FD and C dyes, mixtures thereof; and water insoluble FD and C dyes suspended on alumina hydrate. Sweetening agents include sucrose, lactose, mannitol and artificial sweetening agents such as saccharin, and any number of spray dried flavors. Flavoring agents include natural flavors extracted from plants such as fruits and synthetic blends of compounds which produce a pleasant sensation, such as, but not limited to peppermint and methyl salicylate. Wetting agents include propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate and polyoxyethylene laural ether. Emetic-coatings include fatty acids, fats, waxes, shellac, ammoniated shellac and cellulose acetate phthalates. Film coatings include hydroxyethylcellulose, sodium carboxymethylcellulose, polyethylene glycol 4000 and cellulose acetate phthalate.

[0137] The compound, or pharmaceutically acceptable derivative thereof, could be provided in a composition that protects it from the acidic environment of the stomach. For example, the composition can be formulated in an enteric coating that maintains its integrity in the stomach and releases the active compound in the intestine. The composition may also be formulated in combination with an antacid or other such ingredient.

[0138] When the dosage unit form is a capsule, it can contain, in addition to material of the above type, a liquid carrier such as a fatty oil. In addition, dosage unit forms can contain various other materials which modify the physical form of the dosage unit, for example, coatings of sugar and other enteric agents. The compounds can also be administered as a component of an elixir, suspension, syrup, wafer, sprinkle, chewing gum or the like. A syrup may contain, in addition to the active compounds, sucrose as a sweetening agent and certain preservatives, dyes and colorings and flavors.

[0139] The active materials can also be mixed with other active materials which do not impair the desired action, or with materials that supplement the desired action, such as antacids, H2 blockers, and diuretics. The active ingredient is a compound or pharmaceutically acceptable derivative thereof as described herein. Higher concentrations, up to about 98% by weight of the active ingredient may be included.

[0140] In all embodiments, tablets and capsules formulations may be coated as known by those of skill in the art in order to modify or sustain dissolution of the active ingredient. Thus, for example, they may be coated with a conventional enterically digestible coating, such as phenylsalicylate, waxes and cellulose acetate phthalate. Liquid compositions for oral administration

[0141] Liquid oral dosage forms include aqueous solutions, emulsions, suspensions, solutions and / or suspensions reconstituted from non-effervescent granules and effervescent preparations reconstituted from effervescent granules. Aqueous solutions include, for example, elixirs and syrups. Emulsions are either oil- in-water or water-in-oil.

[0142] Liquid pharmaceutically administrable compositions can, for example, be prepared by dissolving, dispersing, or otherwise mixing an active compound as defined above and optional pharmaceutical adjuvants in a carrier, such as, for example, water, saline, aqueous dextrose, glycerol, glycols, ethanol, and the like, to thereby form a solution or suspension. If desired, the pharmaceutical composition to be administered may also contain minor amounts of nontoxic auxiliary substances such as wetting agents, emulsifying agents, solubilizing agents, pH buffering agents and the like, for example, acetate, sodium citrate, cyclodextrine derivatives, sorbitan monolaurate, triethanolamine sodium acetate, triethanolamine oleate, and other such agents.

[0143] Elixirs are clear, sweetened, hydroalcoholic preparations. Pharmaceutically acceptable carriers used in elixirs include solvents. Syrups are concentrated aqueous solutions of a sugar, for example, sucrose, and may contain a preservative. An emulsion is a two-phase system in which one liquid is dispersed in the form of small globules throughout another liquid. Pharmaceutically acceptable carriers used in emulsions are non-aqueous liquids, emulsifying agents and preservatives. Suspensions use pharmaceutically acceptable suspending agents and preservatives. Pharmaceutically acceptable substances used in non- effervescent granules, to be reconstituted into a liquid oral dosage form, include diluents, sweeteners and wetting agents. Pharmaceutically acceptable substances used in effervescent granules, to be reconstituted into a liquid oral dosage form, include organic acids and a source of carbon dioxide. Coloring and flavoring agents are used in all of the above dosage forms.

[0144] Solvents include glycerin, sorbitol, ethyl alcohol and syrup. Examples of preservatives include glycerin, methyl and propylparaben, benzoic acid, sodium benzoate and ethanol. Examples of non-aqueous liquids utilized in emulsions include mineral oil and cottonseed oil. Examples of emulsifying agents include gelatin, acacia, tragacanth, bentonite, and surfactants such as polyoxyethylene sorbitan monooleate. Suspending agents include sodium carboxymethylcellulose, pectin, tragacanth, Veegum and acacia. Sweetening agents include sucrose, syrups, glycerin and artificial sweetening agents such as saccharin.Wetting agents include propylene glycol monostearate, sorbitan monooleate, diethylene glycol monolaurate and polyoxyethylene lauryl ether. Organic acids include citric and tartaric acid. Sources of carbon dioxide include sodium bicarbonate and sodium carbonate. Coloring agents include any of the approved certified water soluble FD and C dyes, and mixtures thereof. Flavoring agents include natural flavors extracted from plants such fruits, and synthetic blends of compounds which produce a pleasant taste sensation.

[0145] For a solid dosage form, the solution or suspension, in for example propylene carbonate, vegetable oils or triglycerides, is in one embodiment encapsulated in a gelatin capsule. For a liquid dosage form, the solution, e.g., for example, in a polyethylene glycol, may be diluted with a sufficient quantity of a pharmaceutically acceptable liquid carrier, e.g., water, to be easily measured for administration.

[0146] Alternatively, liquid or semi-solid oral formulations may be prepared by dissolving or dispersing the active compound or salt in vegetable oils, glycols, triglycerides, propylene glycol esters (e.g., propylene carbonate) and other such carriers, and encapsulating these solutions or suspensions in hard or soft gelatin capsule shells. Other useful formulations include those set forth in U.S. Patent Nos. RE28,819 and 4,358,603. Briefly, such formulations include, but are not limited to, those containing a compound provided herein, a dialkylated mono- or poly-alkylene glycol, including, but not limited to, 1,2-dimethoxymethane, diglyme, triglyme, tetraglyme, polyethylene glycol-350-dimethyl ether, polyethylene glycol-550-dimethyl ether, polyethylene glycol-750-dimethyl ether wherein 350, 550 and 750 refer to the approximate average molecular weight of the polyethylene glycol, and one or more antioxidants, such as butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), propyl gallate, vitamin E, hydroquinone, hydroxycoumarins, ethanolamine, lecithin, cephalin, ascorbic acid, malic acid, sorbitol, phosphoric acid, thiodipropionic acid and its esters, and dithiocarbamates. Injectables, Solutions and Emulsions

[0147] Parenteral administration, in one embodiment characterized by injection, either subcutaneously, intramuscularly or intravenously is also contemplated herein. Injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution or suspension in liquid prior to injection, or as emulsions. The injectables, solutions and emulsions also contain one or more excipients. Suitable excipients are, for example, water, saline, dextrose, glycerol or ethanol. In addition, if desired, the pharmaceutical compositions to be administered may also contain minor amounts of non-toxic auxiliary substances such as wetting or emulsifying agents, pH buffering agents, stabilizers, solubility enhancers, and other such agents, such as for example, sodium acetate, sorbitan monolaurate, triethanolamine oleate and cyclodextrins.

[0148] Implantation of a slow-release or sustained-release system, such that a constant level of dosage is maintained is also contemplated herein. Briefly, a compound provided herein is dispersed in a solid innermatrix, e.g., polymethylmethacrylate, polybutylmethacrylate, plasticized or unplasticized polyvinylchloride, plasticized nylon, plasticized polyethyleneterephthalate, natural rubber, polyisoprene, polyisobutylene, polybutadiene, polyethylene, ethylene-vinylacetate copolymers, silicone rubbers, polydimethylsiloxanes, silicone carbonate copolymers, hydrophilic polymers such as hydrogels of esters of acrylic and methacrylic acid, collagen, cross-linked polyvinylalcohol and cross-linked partially hydrolyzed polyvinyl acetate, that is surrounded by an outer polymeric membrane, e.g., polyethylene, polypropylene, ethylene / propylene copolymers, ethylene / ethyl acrylate copolymers, ethylene / vinylacetate copolymers, silicone rubbers, polydimethyl siloxanes, neoprene rubber, chlorinated polyethylene, polyvinylchloride, vinylchloride copolymers with vinyl acetate, vinylidene chloride, ethylene and propylene, ionomer polyethylene terephthalate, butyl rubber epichlorohydrin rubbers, ethylene / vinyl alcohol copolymer, ethylene / vinyl acetate / vinyl alcohol terpolymer, and ethylene / vinyloxyethanol copolymer, that is insoluble in body fluids. The compound diffuses through the outer polymeric membrane in a release rate controlling step. The percentage of active compound contained in such parenteral compositions is highly dependent on the specific nature thereof, as well as the activity of the compound and the needs of the subject.

[0149] Parenteral administration of the compositions includes intravenous, subcutaneous and intramuscular administrations. Preparations for parenteral administration include sterile solutions ready for injection, sterile dry soluble products, such as lyophilized powders, ready to be combined with a solvent just prior to use, including hypodermic tablets, sterile suspensions ready for injection, sterile dry insoluble products ready to be combined with a vehicle just prior to use and sterile emulsions. The solutions may be either aqueous or nonaqueous.

[0150] If administered intravenously, suitable carriers include physiological saline or phosphate buffered saline (PBS), and solutions containing thickening and solubilizing agents, such as glucose, polyethylene glycol, and polypropylene glycol and mixtures thereof.

[0151] Pharmaceutically acceptable carriers used in parenteral preparations include aqueous vehicles, nonaqueous vehicles, antimicrobial agents, isotonic agents, buffers, antioxidants, local anesthetics, suspending and dispersing agents, emulsifying agents, sequestering or chelating agents and other pharmaceutically acceptable substances.

[0152] Examples of aqueous vehicles include Sodium Chloride Injection, Ringers Injection, Isotonic Dextrose Injection, Sterile Water Injection, Dextrose and Lactated Ringers Injection. Nonaqueous parenteral vehicles include fixed oils of vegetable origin, olive oil, cottonseed oil, corn oil, sesame oil and peanut oil. Antimicrobial agents in bacteriostatic or fungistatic concentrations must be added to parenteral preparations packaged in multiple-dose containers which include phenols or cresols, mercurials, benzyl alcohol, chlorobutanol, methyl and propyl p-hydroxybenzoic acid esters, thimerosal, benzalkoniumchloride and benzethonium chloride. Isotonic agents include sodium chloride and dextrose. Buffers include phosphate and citrate. Antioxidants include sodium bisulfate. Local anesthetics include procaine hydrochloride. Suspending and dispersing agents include sodium carboxymethylcelluose, hydroxypropyl methylcellulose and polyvinylpyrrolidone. Emulsifying agents include Polysorbate 80 (TWEEN^ 80). A sequestering or chelating agent of metal ions include EDTA. Pharmaceutical carriers also include ethyl alcohol, polyethylene glycol and propylene glycol for water miscible vehicles; and sodium hydroxide, hydrochloric acid, citric acid or lactic acid for pH adjustment.

[0153] The concentration of the pharmaceutically active compound is adjusted so that an injection provides an effective amount to produce the desired pharmacological effect. The exact dose depends on the age, weight and condition of the patient or animal as is known in the art.

[0154] The unit-dose parenteral preparations are packaged in an ampule, a vial or a syringe with a needle. All preparations for parenteral administration must be sterile, as is known and practiced in the art.

[0155] Illustratively, intravenous or intra-arterial infusion of a sterile aqueous solution containing an active compound is an effective mode of administration. Another embodiment is a sterile aqueous or oily solution or suspension containing an active material injected as necessary to produce the desired pharmacological effect.

[0156] Injectables are designed for local and systemic administration. In one embodiment, a therapeutically effective dosage is formulated to contain a concentration of at least about 0.1% w / w up to about 90% w / w or more, in certain embodiments more than 1% w / w of the active compound to the treated tissue(s).

[0157] The compound may be suspended in micronized or other suitable form or may be derivatized to produce a more soluble active product or to produce a prodrug. The form of the resulting mixture depends upon a number of factors, including the intended mode of administration and the solubility of the compound in the selected carrier or vehicle. The effective concentration is sufficient for ameliorating the symptoms of the condition and may be empirically determined. Lyophilized Powders

[0158] Of interest herein are also lyophilized powders, which can be reconstituted for administration as solutions, emulsions and other mixtures. They may also be reconstituted and formulated as solids or gels.

[0159] The sterile, lyophilized powder is prepared by dissolving a compound provided herein, or a pharmaceutically acceptable derivative thereof, in a suitable solvent. The solvent may contain an excipient which improves the stability or other pharmacological component of the powder or reconstituted solution,prepared from the powder. Excipients that may be used include, but are not limited to, dextrose, sorbital, fructose, corn syrup, xylitol, glycerin, glucose, sucrose or other suitable agent. The solvent may also contain a buffer, such as citrate, sodium or potassium phosphate or other such buffer known to those of skill in the art at, in one embodiment, about neutral pH. Subsequent sterile filtration of the solution followed by lyophilization under standard conditions known to those of skill in the art provides the desired formulation. In one embodiment, the resulting solution will be apportioned into vials for lyophilization. Each vial will contain a single dosage or multiple dosages of the compound. The lyophilized powder can be stored under appropriate conditions, such as at about 4 °C to room temperature.

[0160] Reconstitution of this lyophilized powder with water for injection provides a formulation for use in parenteral administration. For reconstitution, the lyophilized powder is added to sterile water or other suitable carrier. The precise amount depends upon the selected compound. Such amount can be empirically determined. Topical Administration

[0161] Topical mixtures are prepared as described for the local and systemic administration. The resulting mixture may be a solution, suspension, emulsions or the like and are formulated as creams, gels, ointments, emulsions, solutions, elixirs, lotions, suspensions, tinctures, pastes, foams, aerosols, irrigations, sprays, suppositories, bandages, dermal patches or any other formulations suitable for topical administration.

[0162] The compounds or pharmaceutically acceptable derivatives thereof may be formulated as aerosols for topical application, such as by inhalation. These formulations for administration to the respiratory tract can be in the form of an aerosol or solution for a nebulizer, or as a microfine powder for insufflation, alone or in combination with an inert carrier such as lactose. In such a case, the particles of the formulation will, in one embodiment, have diameters of less than 50 microns, in one embodiment less than 10 microns.

[0163] The compounds may be formulated for local or topical application, such as for topical application to the skin and mucous membranes, such as in the eye, in the form of gels, creams, solutions and lotions Nasal solutions of the active compound alone or in combination with other pharmaceutically acceptable excipients can also be administered.

[0164] These solutions, particularly those intended for ophthalmic use, may be formulated as 0.01% - 10% (vol%) isotonic solutions, pH about 5-7, with appropriate salts.Compositions for other routes of administration

[0165] Other routes of administration, such as transdermal patches, including iontophoretic and electrophoretic devices, vaginal and rectal administration, are also contemplated herein.

[0166] For example, pharmaceutical dosage forms for rectal administration are rectal suppositories, capsules and tablets for systemic effect. Rectal suppositories are used herein mean solid bodies for insertion into the rectum which melt or soften at body temperature releasing one or more pharmacologically or therapeutically active ingredients. Pharmaceutically acceptable substances utilized in rectal suppositories are bases or vehicles and agents to raise the melting point. Examples of bases include cocoa butter (theobroma oil), glycerin-gelatin, carbowax (polyoxyethylene glycol) and appropriate mixtures of mono-, di- and triglycerides of fatty acids. Combinations of the various bases may be used. Agents to raise the melting point of suppositories include spermaceti and wax. Rectal suppositories may be prepared either by the compressed method or by molding. The weight of a rectal suppository, in one embodiment, is about 2 to 3 gm.

[0167] Tablets and capsules for rectal administration are manufactured using the same pharmaceutically acceptable substance and by the same methods as for formulations for oral administration. Co-administration with other drugs

[0168] In accordance with another aspect of the present invention, it is contemplated that compounds of Formula I as described herein may be administered to a subject in need thereof in combination with medication considered by those of skill in the art to be current standard of care for the condition of interest. Such combinations provide one or more advantages to the subject, e.g., increased efficacy, requiring reduced dosages to achieve similar benefit, obtaining the desired effect in less time, and the like.

[0169] Compounds in accordance with the present invention may be administered as part of a therapeutic regimen with other drugs. It may be desirable to administer a combination of active compounds, for example, for the purpose of treating a particular disease or condition. Accordingly, it is within the scope of the present invention that two or more pharmaceutical compositions, at least one of which contains a compound of Formula (I) according to the present invention, may be combined in the form of a kit suitable for co-administration of the compositions.

[0170] In one embodiment of the methods of the present inventions a compound of Formula I may be administered with a second therapeutic agent. In one embodiment the second therapeutic agent is selected from the group consisting of an anti-hyperglycemic agent, an anti-inflammatory agent, an anti-hypertensiveagent, an agent used to lower cholesterol, a diuretic agent, an anti-fibrotic agent, an agent used to manage heart disease, an anti-angiogenic agent, an antiviral agent and an immunosuppressive agent.

[0171] In another embodiment of the methods of the present inventions a compound of Formula I may be administered with a second therapeutic agent selected from the group consisting of insulin or an insulin analogue; a GLP-1 analogue such as liraglutide, semaglutide, exenatide, lixisenatide and dulaglutide; tirzepatide; survodutide, cotadutide; a biguanide, such as metformin; a thiazolidinedione, such as pioglitazone; a sulfonylurea, such as chlorpropamide, glimepiride, glipizide and tolazamide; dipeptidyl peptidase-4 inhibitors, such as sitagliptin, vildagliptin, saxagliptin and linagliptin; a sodium / glucose cotransporter 2 inhibitor, such as dapagliflozin, empagliflozin, ertugliflozin and canagliflozin; and a thyroid hormone receptor ligand such as resmetirom.

[0172] In another embodiment of the methods of the present inventions a compound of Formula I may be administered with a second therapeutic agent selected from the group consisting of a steroidal anti- inflammatory agent, aspirin, paracetamol, ibuprofen, naproxen, indomethacin, celecoxib, diclofenac and DPP-1 inhibitors like brensocatib.

[0173] In another embodiment of the methods of the present inventions a compound of Formula I may be administered with a second therapeutic agent selected from the group consisting of a calcium channel blocker, such as verapamil, nisoldipine and amlodipine; an angiotensin II receptor antagonist, such as irbesartan, losartan, telmisartan and valsartan; and an inhibitor of angiotensin-converting enzyme, such as benazepril, captopril, enalapril, lisinopril and fosinopril.

[0174] In another embodiment of the methods of the present inventions a compound of Formula I may be administered with a second therapeutic agent selected from the group consisting of a 3-hydroxy-3- methyl-glutaryl-coenzyme A reductase inhibitor, such as atorvastatin, rosuvastatin, simvastatin and pravastatin; a cholesterol absorption inhibitor, such as ezetimibe; a proprotein convertase subtilisin / kexin type 9 inhibitor, such as alirocumab and evolocumab; and a drug used to lower tri-glycerides, such as niacin.

[0175] In another embodiment of the methods of the present inventions a compound of Formula I may be administered with a second therapeutic agent selected from the group consisting of a vasopressin receptor antagonist, such as amphotericin B, tolvaptan and conivaptan; a sodium–proton exchanger antagonist, such as dopamine; a loop diuretic, such as ethacrynic acid, furosemide and torsemide; an osmotic diuretic, such as glucose or mannitol; a potassium-sparing diuretic, such as amiloride, spironolactone and eplerenone; a thiazide, such as bendroflumethiazide and hydrochlorothiazide; and a xanthine, such as theophylline and theobromine.

[0176] In another embodiment of the methods of the present inventions a compound of Formula I may be administered with a second therapeutic agent selected from the group consisting of pirfenidone, nintedanib and bexotegrast.

[0177] In another embodiment of the methods of the present inventions a compound of Formula I may be administered with a second therapeutic agent selected from the group consisting of digoxin, warfarin, a beta-blocker, such as metoprolol, bisoprolol, naldolol, atenolol and propranolol; a vasodilator, such as nitroglycerin, hydralazine, epoprostenol, treprostinil and iloprost; an endothelin receptor antagonist, such as bosentan, ambrisentan and macitentan; and a phosphodiesterase 5 inhibitor, such as sildenafil and tadalafil.

[0178] In another embodiment of the methods of the present inventions a compound of Formula I may be administered with a second therapeutic agent selected from the group consisting of axitinib, bevacizumab, everolimus, lenalidomide, pomalidomide, regorafenib, sorafenib, sunitinib, thalidomide, vandetanib and ziv-aflibercept.

[0179] In another embodiment of the methods of the present inventions a compound of Formula I may be administered with a second therapeutic agent selected from the group consisting of a glucocorticoid, such as prednisone, dexamethasone, and hydrocortisone; a cyostatic agent, such as cyclophosphamide, azathioprine, dactinomycin and methotrexate; an antibody acting directly against an immune response, such as basiliximab, daclizumab and muromonab; agents that act upon the immunophilins, such as cyclosporine, rapamycin, tacrolimus and everolimus; and other immunosuppressant agents, including fingolimod, interferon therapy, infliximab, etanercept and adalimumab.

[0180] In another embodiment of the methods of the present inventions a compound of Formula I may be administered with a second therapeutic agent. In one embodiment the second therapeutic agent is selected from drugs thought to be useful in the treatment of muscular dystrophies, including: prednisone, deflazacort, ataluren, myostatin, eteplirsen, golodirsen, casimersen, idebenone, pamrevlumab, edasalonexent, givinostat, rimeporide and ifetroban.

[0181] In another embodiment of the methods of the present inventions a compound of Formula I may be administered with a second therapeutic agent selected from the group consisting of an antiviral agent such as an antibody, mixture of antibodies and plasma containing antibodies; a nucleoside analogue such as remdesivir, galidesivir, favipiravir and ribavirin; antibiotics, such as doxycycline, azithromycin and ciprofloxacin; chloroquine and hydroxychloroquine.

[0182] When two or more active ingredients are co-administered, the active ingredients may be administered simultaneously, sequentially or separately. In one embodiment the compound of Formula I isco-administered simultaneously with a second therapeutic agent. In another embodiment the compound of Formula I and the second therapeutic agent are administered sequentially. In a further embodiment the compound of Formula I and the second therapeutic agent are administered separately.

[0183] In one embodiment the compound of Formula I is co-administered simultaneously with a second therapeutic agent. In another embodiment the compound of Formula I and the second therapeutic agent are administered sequentially. In a further embodiment the compound of Formula I and the second therapeutic agent are administered separately.

[0184] The invention will now be described in greater detail, by way of illustration only, with reference to the following non-limiting examples. The examples are intended to serve to illustrate the invention and should not be construed as limiting the generality of the disclosure of the description throughout this specification.

[0185] For purposes of this specification, the following abbreviations have the indicated meanings: rt = room temperature min = minute(s) °C = degrees Celsius h = hour(s) NaHMDS = sodium hexamethyldisilazide THF = tetrahydrofuran DMF = dimethylformamide NaOtBu = sodium tertbutoxide TFA = trifluoroacetic acid DIBAL = diisobutylaluminumhydride MTBE = methyl tertbutylether DIPEA = diisopropylethylamine LC-MS = liquid chromatography-mass spectroscopy rpm = revolutions per minute NMR = nuclear magnetic resonance DMSO = dimethylsulfoxide Boc = tert-butyloxycarbonyl HPLC = high-performance liquid chromatography (also, high pressure liquid chromatography) rac = racemic TLC = thin-layer chromatography V = volume(s)Experimental: General methods

[0186] All commercially available solvents and reagents were used as received. Where appropriate, reactions were carried out under an argon atmosphere. Reactions were monitored by either analytical thin- layer chromatography (TLC) or by analytical liquid chromatography−mass spectrometry (LC-MS) recorded on either a Shimadzu LC-MS 2020 instrument or an Agilent LC-MS 1200 instrument using reverse-phase conditions. Purification of intermediates and final compounds was conducted, where necessary, using column chromatography or preparative HPLC. Normal-phase column chromatography was conducted under medium pressure either on silica gel or on prepacked silica gel cartridges using a flash chromatography system (CombiFlash Rf200, Teledyne Isco systems, USA). Reverse-phase column chromatography was conducted under low pressure on prepacked C18 cartridges using a flash chromatography system (Reveleris® X2). Eluents were monitored by UV light (λ = 254 / 280 nm).1H-NMR and19F-NMR spectra were recorded using either a Bruker 300 MHz NMR spectrometer or a Bruker Avance III plus 400 MHz NMR spectrometer. Chemical shifts (δ) are reported as parts per million (ppm) relative to tetramethylsilane (TMS; internal standard). The following abbreviations are used for multiplicities: s = singlet; br s = broad singlet; d = doublet; t = triplet; q = quartet; p = pentet; m = multiplet; and br m = broad multiplet. Low resolution mass spectra (MS) were obtained as electrospray – atmospheric pressure ionization (ES-API) mass spectra on a Shimadzu LC-MS 2020 or an Agilent LC-MS 1200 instrument. All animal experiments were conducted in compliance with institutional guidelines and approval from local ethics committees. EXAMPLE 1

[0187] Synthesis of (E)-3-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)benzyl)-2-thioxo-1,2,3,7-tetrahydro- 6H-purin-6-one hydrochloride (Compound 1)Procedure A: Preparation of tert- 3- 1H- 1- (Int-1)

[0188] To a vessel charged with acetonitrile (754 L) under nitrogen was added sequentially β-alanine tert-butyl ester hydrochloride (75 kg), 2,5-hexanedione (97.5 L) and potassium carbonate (75 kg). Theresulting mixture was heated at 75 °C for 2 h and then stirred for a further 4 h. The reaction mixture was cooled to 55 °C and then concentrated in vacuo to ~290 L. Water (1200 L) was then added to the residue at 55 °C. The mixture was stirred at 55 °C for 1 h, and then cooled at 10 °C for 2 h. Stirring was continued at 10 °C for 10 h. The slurry was then spun down in a centrifuge and the liquid phase was then removed. The resulting “cake” was rinsed with further water (150 L x 3), spinning down (centrifuge) and removing the liquid phase after each rinsing cycle. After final removal of the liquid phase, the wet “cake” was dried under vacuum at 40 °C for 24 h to afford Int-1 (79.4 kg, 85%).1H-NMR (300 MHz, CDCl3) δ ppm: 5.78 (s, 2H), 4.02 – 4.07 (m, 2H), 2.50 – 2.56 (m, 2H), 2.25 (s, 6H), 1.47 (s, 9H). Procedure B: Preparation of 1-(tert-butyl) 3-ethyl 2-(difluoromethyl)-2-((2,5-dimethyl-1H-pyrrol-1- yl)methyl)malonate (Int-2)

[0189] This process was performed on two batches of Int-1 (39.5 kg x 2). The batches were combined at a late stage.

[0190] To vessel (A), charged with THF (277 L) under nitrogen was added NaHMDS (2.0 M; 245 kg) over 20 min. The resulting mixture was cooled to -58 °C. To vessel (B), charged with THF (198 L) under nitrogen at -50 °C was added Int-1 (39.5 kg). The resulting mixture was stirred at -50 °C for 25 min. The contents of vessel (B) was then transferred to vessel (A) over 30 min at -58 °C. To vessel (A) was then added ethyl chloroformate (17.4 L) over 1 h during which time an exotherm was observed. The resulting mixture was stirred at -58 °C for 1 h. To this was added NaOtBu (15.3 kg) over 15 min. Chlorodifluoromethane (17.8 kg) was then introduced slowly at -20 °C. The mixture was stirred at -30 °C for 2 h. The vessel was then charged with further chlorodifluoromethane (7.00 kg) slowly at -20 °C. Stirring was continued for a further 2 h at -20 °C. The reaction vessel was charged with water (356 L) followed by citric acid (79 kg). The resulting mixture was stirred for 30 min and the organic layer separated. NaCl solution (10%, 342.4 kg) was added and the mixture stirred for 1 h. The organic layer was separated and concentrated in vacuo. Residue was dissolved in n-heptane (584.8 kg) and silica gel added. Stirred for 30 min and then filtered through Celite®, washing with n-heptane (434 kg). Combined filtrate was concentrated in vacuo and the residue thus obtained recrystallized from EtOH:H2O (1:1) to afford Int 2 as a light yellow solid (91.0 kg, 74.5%).1H-NMR (300 MHz, CDCl3) δ ppm: 6.29 (t, J = 54.6 Hz, 1H), .72 (s, 2H), 4.56 – 4.69 (m, 2H), 4.12 – 4.23 (m, 1H), 3.98 – 4.08 (m, 1H), 2.20 (s, 6H), 1.36 (s, 9H), 1.23 (t, J = 7.2 Hz, 3H).Procedure C: Preparation of ethyl (E)-2-((2,5-dimethyl-1H-pyrrol-1-yl)methyl)-3-fluoroacrylate (Int-3)

[0191] To reaction vessel (A), charged with TFA (279 L) at 10 °C under nitrogen was added Int-2 (91 kg). The resulting mixture was heated to 35 °C, left to stir for 13 h and then cooled to 5 °C. To reaction vessel (B) was added triethylamine (620 L) followed by n-heptane (532 L). The resulting solution was cooled to 5 °C. The content of reactor (A) was then transferred to reactor (B) over 2 h. The resulting mixture in reactor vessel (B) was heated to 40 °C and stirring was continued for 10 h. The reaction mixture was cooled to 10 °C and then washed with water (910 L). The organic phase was separated and washed with a solution of citric acid (91 kg) in water (455 L) followed by a solution of NaCl (73 kg) in water (410 L). To the organic phase was added silica gel (91 kg) and the resulting slurry was stirred for 30 min. The slurry was then filtered and the filter “cake” was washed with n-heptane. The filtrate was concentrated in vacuo and the resulting residue was taken up in toluene (520 L). After further concentration to 650 – 750 L, the resulting solution of crude Int-3 was used in the subsequent step.1H-NMR (300 MHz, CDCl3) δ ppm: 7.60 (d, J = 80.1 Hz, 1H), 5.77 (s, 2H), 4.67 – 4.68 (m, 2H), 4.20 (q, J = 7.2 Hz, 2H), 2.26 (s, 6H), 1.27 (t, J = 7.2 Hz, 3H). Procedure D: Preparation of (E)-2-((2,5-dimethyl- pyrrol-1-yl)methyl)-3-fluoroprop-2-en-1-ol (Int-4)

[0192] To the solution of Int-3 in toluene (520 L), prepared in the previous step, under nitrogen at -55 °C was added DIBAL (1.5 M in toluene; 199.2 kg) dropwise over 4 h. The resulting mixture was stirred at -45 °C for 1 h. In-process control analysis after this time indicated incomplete conversion. A further amount of DIBAL (1.5 M in toluene; 15 kg) was added dropwise over 40 min at -55 °C. The mixture was then stirred at -45 °C for a further 30 min and then warmed to 10 °C. To quench the reaction, a cold (0 °C) solution of citric acid (64 kg) in water (320 L) was added slowly over 2 h. The organic phase was separated and washed with two solutions of NaCl (32 kg) in water (160 L). Silica gel (32 kg) was added to the organic phase and the resulting slurry was stirred for 30 min. The slurry was filtered through a pad of silica gel (160 kg), and the filter “cake” was washed with toluene (296 L). The filtrate was then concentrated to ~50 L. To the residue was added n-heptane (319 L), and the resulting mixture was cooled to 0 °C and stirred for 10 h. The solid was then filtered and dried to afford Int-4 (11.4 kg, 24% over two steps) as an off-white solid.1H-NMR (400 MHz, CDCl3) δ ppm: 6.70 (d, J = 83.4 Hz, 1H), 5.80 (s, 2H), 4.63 (t, J = 1.2 Hz, 2H), 3.76 (t, J = 4.0 Hz, 2H), 2.23 (s, 6H).Procedure E: Preparation of tert-butyl (E)-(3-fluoro-2-(hydroxymethyl)allyl)carbamate (Int-5)

[0193] To a reaction vessel charged with Int-4 (2.0 kg), hydroxylamine hydrochloride (3.80 kg) and water (8.00 L) was added a solution of potassium hydroxide (1.20 kg) in water (10.0 L). The resulting mixture was heated to reflux and stirred for 18 h. After cooling to rt, the reaction mixture was filtered and the filter “cake” washed with water (6.0 L). To the filtrate was then added sodium carbonate (3.46 kg) and the resulting mixture was stirred for 30 min. Acetone (9.1 L) was added dropwise at 20 °C, and the mixture was stirred for 2 h. Finally, a solution of di-tert-butyldicarbonate (2.37 kg) in MTBE, (10 L) was added and stirring was then continued for a further 18 h at 15 °C. The organic layer was then separated and washed with aqueous NaOH (1.0 M; 16 L x 4). To the organic layer was added silica gel (2.0 kg) and the resulting slurry was stirred for 30 min. The mixture was then filtered and the filter “cake” was washed with MTBE (10 L). The filtrate was concentrated in vacuo to afford Int-5 (2.0 kg, 89%).1H-NMR (400 MHz, CDCl3) δ ppm: 6.78 (d, J = 81.2 Hz, 1H), 4.86 (br. s, 1H), 4.00 – 3.96 (m, 4H), 1.45 (s, 9H). Procedure F: Preparation of tert-butyl (bromomethyl)-3-fluoroallyl)carbamate (Int-6)

[0194] To a vessel charged with Int-5 (4.0 kg), DIPEA (10.2 L) and THF (40.0 L) at 5 °C was added a solution of methanesulfonic anhydride (5.78 kg) in THF (20.0 L). The resulting mixture was stirred for 30 min. To this was added lithium bromide (8.47 kg) and stirring was continued at 5 °C for 2 h. Water (20.0 L) was then added dropwise to the reaction mixture. The organic layer was separated and concentrated to ~8.0 L. To this was added ethyl acetate (24 L) and water (20 L). The organic layer was again separated and concentrated to ~12 L. Heptane (40 L) and silica gel (2.0 kg) was added and the resulting slurry was stirred for 30 min. The slurry was then filtered and the filter “cake” was washed with n-heptane (20 L). The filtrate was concentrated to dryness in vacuo. Further n-heptane was added and stirring was continued at 5 °C for 1 h. The resulting solid was collected by filtration and dried under vacuum at 30 °C to afford Int-6 (3.20 kg, 61%).1H-NMR (400 MHz, CDCl3) δ ppm: 6.78 (d, J = 81.2 Hz, 1H), 4.68 (br.s, 1H), 4.00 (d, J = 4.4 Hz, 2H), 3.96 (d, J = 3.2 Hz, 2H), 1.45 (s, 9H).Procedure G: Preparation of 4-((3-hydroxybenzyl)amino)-1H-imidazole-5-carboxamide (Int-7)

[0195] To a stirring solution 3-hydroxybenzaldehyde (20.0 kg) in MeOH (240 kg) under at N2 at 20 °C was added 4-amino-1H-imidazole-5-carboxamide (20.0 kg) and acetic acid (9.40 kg). The resulting mixture was stirred at 20 °C for 8 h. The reaction mixture was filtered and the solid was washed with MeOH (40.0 kg). The wet “cake” was transferred to a second reaction vessel. To this vessel was added MeOH (200 kg) and THF (228 kg) followed by Pd / C (3.00 kg). The resulting slurry was stirred at 20 °C for 15 min before introduction of H2, keeping the pressure at 0.3 MPa. The reaction mixture was stirred at 20 °C under an atmosphere of H2for 20 h. The reaction mixture was filtered, and the filtrate was concentrated to ~3 V. The resulting solution was used directly in the next step.1H NMR (300 MHz, DMSO-d6) δ pm: 11.96 (s, 1H), 9.40 (s, 1H), 7.24 (s, 1H), 7.08 (t, J = 8.0 Hz, 1H), 6.86 – 6.66 (m, 4H), 6.61 (dd, J = 8.2, 2.3 Hz, 1H), 6.37 (s, 1H), 4.34 (d, J = 6.7 Hz, 2H). Procedure H: Preparation of 4-(3-benzoyl-1-(3-hydroxybenzyl)thioureido)-1H-imidazole-5-carboxamide (Int-8)

[0196] To a stirring solution of Int-7 (as a solution progressed from Procedure G) in MeOH (127 kg) and CH2Cl2 (424 kg) at 20 °C was added benzoyl isothiocyanate (37.5 kg). The resulting mixture was stirred at 20 °C for 10 h. The reaction mixture was filtered and the solid “cake” was washed with CH2Cl2 (84.8 kg). The solid was then dried under vacuum at 35 °C for 12 h to afford Int-8 (55.0 kg, 88% over three steps). MS Found: 396.06 [M+H]+. Procedure I: Preparation of purin-6-one (Int-9)

[0197] To a stirring solution of Int-8 (55.0 kg) in MeOH (654 kg) under N2 was added aqueous potassium hydroxide (5.0 M; 385 L). An exotherm was observed during addition. The resulting mixture was stirred at 25 °C for 26 h. The pH of the reaction mixture was adjusted to ~6.5 by the addition of aqueous HCl (6.0 M; 400 kg) at a rate that kept the reaction temperature at ~25 °C. The mixture was stirred at 25 °C for a further 30 min. The reaction mixture was filtered, and the filter “cake” was washed with water (130.8 L x 2) followed by MeOH (55.1 L). The solid was then dried at 50 °C for 12 h to afford Int-9 (32.0 kg, 84%).1H-NMR (300 MHz, DMSO-d6) δ ppm: 9.32 (s, 1H), 8.10 (d, J = 17.1 Hz, 1H), 7.08 (t, J = 7.8 Hz, 1H), 6.77 (d, J = 7.6 Hz, 1H), 6.69 (t, J = 2.0 Hz, 1H), 6.65 – 6.58 (m, 1H), 5.64 (s, 2H). Procedure J: Preparation of 3-(3-hydroxybenzyl)-7-(4-methoxybenzyl)-2-((4-methoxybenzyl)thio)-3,7- dihydro-6H-purin-6-one (Int-10)

[0198] To a suspension of Int-9 (32.0 kg) and K2CO3(64.5 kg) in DMF (160 kg) at 25 °C was slowly added para-methoxybenzyl chloride (PMBCl) (43.8 kg). The resulting mixture was left to stir at 25 °C for 8 h. The reaction mixture was diluted with ethyl acetate (160 kg) and water (320 kg). The resulting solid was filtered and the filter “cake” was washed with ethyl acetate (32 kg). A suspension of the crude solid and THF (160 kg) was heated at reflux for 1 h. The solution (not completely homogeneous) was then cooled to 5 °C and stirring was continued for a further 1 h. The resulting solid was filtered and then dried under vacuum at 55 °C to afford Int-10 (27.0 kg, 45%).1H-NMR (300 MHz, DMSO-d6) δ ppm: 9.42 (s, 1H), 8.32 (s, 1H), 7.45 – 7.38 (m, 2H), 7.38 – 7.30 (m, 2H), 7.14 – 7.03 (m, 1H), 6.96 – 6.89 (m, 2H), 6.89 – 6.81 (m, 2H), 6.64 (dt, J = 7.6, 2.3 Hz, 2H), 6.56 (t, J = 2.0 Hz, 1H), 5.51 (s, 2H), 5.32 (s, 2H), 4.42 (d, J = 2.5 Hz, 2H), 3.73 (s, 3H), 3.73 (s, 3H). Procedure K: Preparation of tert-butyl (E)-(3-fluoro-2-((3-((7-(4-methoxybenzyl)-2-((4- - - (Int-11)

[0199] To a stirring solution of Int-10 (17.7 kg) in DMF (88.0 L) at 20 °C was added Cs2CO3(16.8 kg). After stirring for 5 min, Int-6 (9.7 kg) was added and the resulting suspension was stirred at 20 °C for 5 h. The reaction mixture was diluted with water (176 L) and isopropylacetate (176 L). The organic phase was separated, washed with further water (88 L x 2) and then concentrated to ~70 L. The product was thenobtained in pure form through a series of precipitations as follows. The organic phase (~70 L) was warmed to 35 °C before the addition of MTBE (211 L). The mixture was then cooled to 5 °C and stirred for 3 h. The solid was then filtered and tested for purity. The level of residual Int-6 was found to be 130 ppm. To the solid was added MTBE (88 L), and the resulting suspension was heated, with stirring, at 55 °C for 2 h. After addition of n-heptane (88 L), the mixture was cooled to 5 °C and the solid was then filtered. The level of residual Int-6 in the filtered solid was found to be below 3 ppm. The solid was dried under vacuum at 45 °C for 20 h to afford Int-11 (19.5 kg, 81%).1H-NMR (300 MHz, CDCl3) δ ppm: 7.57 (s, 1H), 7.46 – 7.39 (m, 2H), 7.39 – 7.33 (m, 2H), 7.22 (t, J = 7.9 Hz, 1H), 6.94 – 6.83 (m, 8H), 5.58 (s, 2H), 5.42 (d, J = 7.1 Hz, 2H), 4.53 (s, 2H), 4.42 – 4.32 (m, 2H), 4.02 – 3.91 (m, 2H), 3.82 (s, 3H), 3.81 (s, 3H), 1.42 (s, 9H). Procedure L: Preparation of (E)-3-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)benzyl)-2-thioxo-1,2,3,7- tetrahydro-6H-purin-6-one hydrochloride (Compound 1)

[0200] To a reaction vessel charged with 1,2-dimethoxybenzene (7.70 kg) and TFA (90.0 kg) at 20 °C was added Int-11 (19.5 kg) portion-wise. An exotherm with concomitant gas release was observed. The resulting mixture was stirred at 20 °C for 1 h. The reaction mixture was then concentrated to remove isobutene. To the residue was added TFA (90.0 kg) and the resulting mixture was stirred at 75 °C for 5 h. The reaction mixture was then concentrated to ~59 L (3 V). To remove residual TFA (through co- evaporation), ethyl acetate (50 L) was added and the mixture was concentrated to ~59 L. This process was then repeated with further ethyl acetate (50 L). To the solution of crude product (59 L, 3 V) was added further ethyl acetate (50 L) and the resulting solution was cooled to 10 °C and stirring was then continued for 10 h. The solid was then filtered and dried to afford the TFA salt (11.5 kg). To a stirred mixture of this material in THF (138 L) was added HCl (4.0 M in ethyl acetate; 35.0 L). The resulting mixture was then warmed to 55 °C to achieve a clear, homogeneous solution. After cooling to 35 °C, the mixture was filtered through filter paper. The filtrate was concentrated to ~35 L. THF (55 L) was added and the solution was again concentrated to ~35 L. This process (to remove residual water) was repeated twice more and, after the final concentration step, the volume was ~55 L. The solid was then filtered and the filter “cake” was washed with THF (12 L). The solid was then dried under vacuum at 55 °C for 40 h. The solid was then suspended in ethanol (10 V) and stirred at 60 °C for 2 h. The solid was then filtered and the filter “cake” was washed with ethanol (1 V) and then dried at 60 °C for 60 h to afford Compound 1 (8.4 kg, 76%).1H- NMR (400 MHz, D2O) δ ppm: 7.94 (s, 1H), 7.11 (t,1H, J = 8 Hz); 6.92 (d, 1H, J = 80.8 Hz), 6.7-6.8 (m, 3H), 5.50 (s, 2H), 4.43 (d, 2H, J = 3.2 Hz), 3.70 (d, 2H, J = 1.6 Hz);19F-NMR (400 MHz, D2O) ^ ppm: - 120.96; MS Found: 362 [M+H]+.EXAMPLE 2

[0201] Synthesis of (Z)-3-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)benzyl)-2-thioxo-1,2,3,7-tetrahydro- 6H-purin-6-one hydrochloride (Compound 2)Procedure M: Preparation of (Z)-3-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)benzyl)-2-thioxo-1,2,3,7- tetrahydro-6H-purin-6-one; trifluoroacetic acid salt (Compound 2)

[0202] Step 1: To a solution of Int-10 (1.60 g) in DMF (16.0 mL) was added Cs2CO3(1.52 g), the mixture was stirred at 10 - 15 °C for 5 min. To this was added Int-5 (as a mixture of isomers: Z / E = 10:90; 0.84 g) was added and the resulting mixture was stirred at 10 – 15 °C for 2 h. The reaction mixture was partitioned between water (32 mL) and ethyl acetate (32 mL). The organic layer was dried and concentrated in vacuo. To the residue was added MTBE (32 mL) and the resulting solid was filtered to afford crude product (1.90 g). This material was used in the next step without purification.

[0203] Step 2: A solution of the crude product from step 1 (1.90 g) in TFA (10.0 mL) was heated at reflux for 1 h. After cooling the reaction mixture to rt, ethyl acetate (19.0 mL) was added and the resulting solid was filtered. Purification was performed by prep-HPLC [Waters Xbridge-prep-C18 (150 mm x 19 mm), A=0.1% HCl in water; B=MeCN gradient, 5 - 95% B over 15 min, Flow: 15.0 mL / min] to afford Compound 2 (120 mg).1H-NMR (400 MHz, DMSO-d6) δ ppm: 8.16 (s, 1H), 7.25 (t, J = 7.9 Hz, 1H), 7.17 (d, J = 81.0 Hz, 1H), 6.99 – 6.83 (m, 3H), 5.70 (s, 2H), 4.65 (d, J = 2.6 Hz, 2H), 3.52 (d, J = 3.0 Hz, 2H).; MS Found: 362 [M+H]+.EXAMPLE 3

[0204] Synthesis of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)-5-fluorobenzyl)-7-methyl-2-thioxo- 1,2,3,5-tetrahydro-4H-pyrrolo[3,2-d]pyrimidin-4-one hydrochloride (Compound 3)Procedure N: Preparation of tert-butyl (E)-(3-fluoro-2-((3-fluoro-5-formylphenoxy)methyl)allyl)- carbamate (Int-12)

[0205] To a solution of 3-fluoro-5-hydroxybenzaldehyde (0.20 g) in DMF (3.0 ml) was added Int-6 (0.40 g) and NaOtBu (0.17 g). The resulting mixture was stirred at rt for 2 h. The reaction was partitioned between MTBE (20 mL) and water (20 mL). The aqueous layer was then extracted with further MTBE (20 mL). The combined organics were washed with water, brine and then dried over Na2SO4. Concentration of the solvent in vacuo afforded crude Int-12. This material was used directly in the next step without further purification. MS Found: 328 [M+H]+. Procedure O: Preparation of ethyl (E)-3-((3-((2-(((tert-butoxycarbonyl)amino)methyl)-3-fluoroallyl)oxy)- 5-fluorobenzyl)amino)-4-methyl-1H-pyrrole-2-carboxylate (Int-13)

[0206] To a stirred solution of crude Int-12 in CH2Cl2 (10 mL) was added ethyl 3-amino-4-methyl-1H- pyrrole-2-carboxylate (0.24 g) and acetic acid (1 drop). The resulting mixture was stirred at rt overnight. To the mixture was added ethanol (5.0 mL) followed by NaBH3CN (0.26 g) and stirring was continued at rt for 2 h. The reaction mixture was concentrated in vacuo. The residue taken up in ethyl acetate (20 mL), and then washed with water (20 mL). The organic layer was dried over Na2SO4 and then concentrated in vacuo. Purification was performed using preparative-HPLC [Gemini nx C18 reverse-phase column (5 μm, 19 x 150 mm) with a linear gradient of the binary solvent system 10 - 80% B; A: H2O (0.1% NH4HCO3),B: acetonitrile, with a flow rate of 15mL / min and UV detection at 214 nm; GT 12 min] to afford Int-13 (0.33 g, 49%).1H-NMR (300 MHz, CDCl3) δ ppm: 8.80-6.60 (m, 1H), 6.68-6.66 (m, 2H), 6.62-6.47 (m, 2H), 4.82 (bs, 1H), 4.83 (s, 2H), 4.41-4.385 (m, 4H), 4.29-4.24 (m, 2H), 3.97-3.96 (m, 2H), 2.04 (s, 3H), 1.42 (s, 9H), 1.33-1.28 (m, 3H); MS Found: 480 [M+H]+. Procedure P: Preparation of tert-butyl (E)-(3-fluoro-2-((3-fluoro-5-((7-methyl-4-oxo-2-thioxo-2,3,4,5- tetrahydro-1H-pyrrolo[3,2-d]pyrimidin-1-yl)methyl)phenoxy)methyl)allyl)carbamate (Int-14)

[0207] To a stirred solution of Int-13 (0.33 g) in CH2Cl2 (10 mL) at rt was added a solution of benzoyl isothiocyanate (0.22 g) in CH2Cl2 (1.0 mL) drop-wise. The resulting mixture was stirred at rt for 6 h. The reaction mixture was concentrated in vacuo. The residue was then dissolved in MeOH (15 mL). To this was added Cs2CO3 (0.56 g), and the resulting suspension was stirred at 60 °C for 3 h. The reaction mixture was then concentrated to dryness. MTBE (10 mL) and H2O (15 mL) were added and the aqueous layer was acidified to pH = 5 with acetic acid. To the organic layer was added petroleum ether (30 mL). The resulting solid was filtered and washed with water and further petroleum ether. The solid was then slurried in MeOH (0.5 mL), MTBE (1 mL) and petroleum ether (10 mL), filtered and then dried under vacuum to afford Int- 14 (0.22 g, 65%).1H-NMR (400 MHz, DMSO-d6) δ ppm: 12.39 (s, 1H), 12.26 (s, 1H), 7.14-6.75 (m, 4H), 6.52-6.48 (m, 2H), 4.38 (s, 2H), 3.71 (s, 2H), 1.96 (s, 3H), 1.29 (s, 9H). MS Found: 493 [M+H]+. Procedure Q: Preparation of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)-5-fluorobenzyl)-7-methyl-2- thioxo-1,2,3,5-tetrahydro-4H-pyrrolo[3,2-d]pyrimidin-4-one hydrochloride (Compound 3)

[0208] To a stirred solution of Int-14 (0.15 g) in CH2Cl2 (3.0 mL) at rt was added TFA (1.5 mL). The resulting mixture was left to stir at rt for 1 h. The solvent and TFA were removed in vacuo, and the residue was purified by preparative-HPLC [Gemini NX C18 reverse-phase column (5 μm, 21.2 x 150 mm) with a linear gradient of the binary solvent system 10 - 50% B, A: H2O (0.1% NH4HCO3), B: acetonitrile, with a flow rate of 20 mL / min and UV detection at 214 nm over a period of12 min]. The purified material was then lyophilized. The white solid was dissolved in MeCN (3 mL) and H2O (2 mL), acidified with 1 M HClto pH = 1 and then lyophilized again to afford Compound 3 (67 mg, 60%) as a white solid.1H-NMR (400 MHz, DMSO-d6) δ ppm: 12.41 (s, 1H), 12.29 (s, 1H), 8.21 (s, 3H), 7.40-7.19 (m, 1H), 7.15 (d, J = 2.4 Hz, 1H), 6.83 (d, J = 10.4 Hz, 1H), 6.57 (d, J = 9.6 Hz, 1H), 6.51 (s, 1H), 5.33 (bs, 1H), 4.58 (s, 2H), 3.57 (s, 2H), 3.37(s, 2H), 1.96 (s, 3H); MS Found: 393 [M+H]+. EXAMPLE 4

[0209] Synthesis of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)-2-fluorobenzyl)-7-methyl-2-thioxo- 1,2,3,5-tetrahydro-4H-pyrrolo[3,2-d]pyrimidin-4-one hydrochloride (Compound 4)

[0210] Preparation of tert-butyl (E)-(3-fluoro-2-((2-fluoro-3-formylphenoxy)methyl)allyl)-carbamate (Int-15)

[0211] Int-15 was prepared according to the experimental protocol described in procedure N from 2- fluoro-3-hydroxybenzaldehyde and Int-6 as a light yellow oil; (340 mg, 99%). MS Found: 328 [M+H]+.

[0212] Preparation of ethyl (E)-3-((3-((2-(((tert-butoxycarbonyl)amino)methyl)-3-fluoroallyl)oxy)-2- fluorobenzyl)amino)-4-methyl-1H-pyrrole-2-carboxylate (Int-16)

[0213] Int-16 was prepared by modification of the experimental protocol described in Procedure O from Int-15 and ethyl 3-amino-4-methyl-1H-pyrrole-2-carboxylate. No EtOH was used and the reaction mixture was adjusted to pH~8 with aqueous NaHCO3prior to workup. Purification was performed using reverse- phase chromatography (80% CH3CN in H2O) to afford Int-16 as a brown oil (233 mg, 50%).1H-NMR (400 MHz, CDCl3) δ ppm: 8.05 (bs, 1H), 7.04 - 6.96 (m, 2H), 6.90 - 6.85 (m, 1H), 6.74 (d, J = 89.6 Hz, 1H), 6.52 (d, J = 2.4 Hz, 1H), 4.89 (s, 1H), 4.48 (s, 4H), 4.27 (q, J = 7.2 Hz, 2H), 4.01 (d, J = 4.0 Hz, 2H), 2.09 (s, 3H), 1.42 (s, 9H), 1.33-1.30 (m, 3H); MS Found: 480 [M+H]+.

[0214] Preparation of tert-butyl (E)-(3-fluoro-2-((2-fluoro-3-((7-methyl-4-oxo-2-thioxo-2,3,4,5- tetrahydro-1H-pyrrolo[3,2-d]pyrimidin-1-yl)methyl)phenoxy)methyl)allyl)carbamate (Int-17)

[0215] Int-17 was prepared according to the experimental protocol described in Procedure P from Int-16; (147 mg, 64%).1H-NMR (400 MHz, DMSO-d6) δ ppm: 12.41 (s, 1H), 12.26 (br.s, 1H), 7.16 - 6.95 (m, 5H), 6.34 (t, J = 6.8 Hz, 1H), 4.51 (s, 2H), 3.77 (d, J = 4.4 Hz, 2H), 1.94 (s, 3H), 1.34 (s, 9H); MS Found: 493 [M+H]+.

[0216] Preparation of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)-2-fluorobenzyl)-7-methyl-2-thioxo- 1,2,3,5-tetrahydro-4H-pyrrolo[3,2-d]pyrimidin-4-one hydrochloride (Compound 4)

[0217] Compound 4 was prepared according to the experimental protocol described in Procedure Q from Int-17. Purification was performed using preparative-HPLC [Gilson-2 Xbridge C18 reverse-phase column (5 μm, 19 x 150 mm) with a linear gradient of the binary solvent system 5 - 60% B, A: H2O (0.1% NH4HCO3), B: acetonitrile, with a flow rate of 15 mL / min and UV detection at 214 nm; GT 12 min] to afford Compound 4 (58.0 mg, 50%).1H-NMR (400 MHz, MeOH-d4) δ ppm: 7.23-6.92 (m, 4H), 6.38 (t, J = 6.8 Hz, 1H), 5.88 (brs, 2H), 4.61 (d, J = 2.8 Hz, 2H), 3.76 (s, 2H), 1.94 (s, 3H); MS Found: 393 [M+H]+. EXAMPLE 5

[0218] Synthesis of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)-2-fluorobenzyl)-2-thioxo-1,2,3,5- tetrahydro-4H-pyrrolo[3,2-d] 5)

[0219] Preparation of ethyl (E)-3-((3-((2-(((tert-butoxycarbonyl)amino)methyl)-3-fluoroallyl)oxy)-2- fluorobenzyl)amino)-1H-pyrrole-2-carboxylate (Int-18)

[0220] Int-18 was prepared according to the experimental protocol outlined in Procedure O from Int-15 and ethyl 3-amino-1H-pyrrole-2-carboxylate. No ethanol was used and the reaction mixture was adjusted to pH~7 with aqueous NaHCO3prior to workup. Purification was performed using reverse-phase chromatography (80% CH3CN in H2O) to afford Int-18 as a brown oil (250 mg, 66%).1H-NMR (400 MHz, CDCl3) δ ppm: 8.13 (brs, 1H), 7.00-6.64 (m, 5H), 5.71 (t, J = 3.2 Hz, 1H), 4.88 (s, 1H), 4.49 (d, J = 3.2 Hz, 2H), 4.42 (d, J = 5.6 Hz, 2H), 4.30 (d, J = 6.0 Hz, 2H), 4.02 (d, J = 5.2 Hz, 2H), 1.42 (s, 9H), 1.34 (t, J = 7.2 Hz, 3H); MS Found: 466 [M+H]+.

[0221] Preparation of tert-butyl (E)-(3-fluoro-2-((2-fluoro-3-((4-oxo-2-thioxo-2,3,4,5-tetrahydro-1H- pyrrolo[3,2-d]pyrimidin-1-yl)methyl)phenoxy)methyl)allyl)carbamate (Int-19)

[0222] Int-19 was prepared by modification of the experimental protocol detailed in Procedure P from Int-18. In this instance, sodium hydroxide was used in place of Cs2CO3, and stirring was continued at rt for 3 d. After workup Int-18 was afforded as a white solid (200 mg, 78%).1H-NMR (400 MHz, DMSO-d6) δ ppm: 12.49 (s, 1H), 12.37 (s, 1H), 7.32 (d, J = 2.8 Hz, 1H), 7.16-6.95 (m, 4H), 6.56 (t, J = 6.8 Hz, 1H), 6.10 (d, J = 2.0 Hz, 1H), 5.70 (s, 2H), 4.50 (d, J = 2.8 Hz, 2H), 3.77 (d, J = 4.8 Hz, 2H), 1.34 (s, 9H); MS Found: 479 [M+H]+.

[0223] Preparation of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)-2-fluorobenzyl)-2-thioxo-1,2,3,5- tetrahydro-4H-

[0224] Compound 5 was prepared according to the experimental protocol outlined in Procedure Q from Int-19. The product was purified using preparative-HPLC [Gilson Xbridge C18 reverse-phase column (5 μm, 19 x 150 mm) with a linear gradient of the binary solvent system 5 - 60% B, A: H2O (0.1% NH4HCO3), B: acetonitrile, with a flow rate of 15 mL / min and UV detection at 214 nm; GT 12 min] to afford Compound 5 (69.0 mg, 61%).1H-NMR (400 MHz, DMSO-d6) δ ppm: 12.54 (s, 1H), 12.39 (s, 1H), 8.47 (s, 2H), 7.43-7.23 (m, 2H), 7.18-7.14 (m, 1H), 7.03 (t, J = 8.0 Hz, 1H), 6.59 (t, J = 6.8 Hz, 1H), 6.12 (d, J = 2.8 Hz, 1H), 5.71 (s, 2H), 4.74 (s, 2H), 3.60 (s, 2H); MS Found: 379 [M+H]+. EXAMPLE 6

[0225] Synthesis of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)-5-fluorobenzyl)-2-thioxo-1,2,3,5- tetrahydro-4H-pyrrolo[3,2-d]pyrimidin-4-one hydrochloride (Compound 6)Procedure R: Preparation of ethyl (E)-3-((3-((2-(((tert-butoxycarbonyl)amino)methyl)-3-fluoroallyl)oxy)- 5-fluorobenzyl)amino)-1H-pyrrole-2-carboxylate (Int-20)

[0226] To a stirred solution of Int-12 (317 mg) and ethyl 3-amino-1H-pyrrole-2-carboxylate (150 mg) in MeOH (5.0 mL) at rt was added NaBH3CN (91.4 mg). The resulting mixture was stirred at rt overnight. TLC analysis after this time showed unreacted aldehyde and amine starting material. Additional quantities of NaBH3CN (91.4 mg) and acetic acid (83 uL) were added and stirring was continued for a further 2 h. Water (10 mL) was added and the aqueous layer was extracted with CH2Cl2(10 mL x 2). The combined organic layer was concentrated and adsorbed onto silica gel. Purification was performed over silica gel (CombiFlash®), eluting with 10 – 50% ethyl acetate in hexanes to afford Int-20 (150 mg) as a mixture of desired product:unreacted amine (1.0:1.4). This mixture was used in the next step without purification.Procedure S: Preparation of ethyl (E)-3-(3-benzoyl-1-(3-((2-(((tert-butoxycarbonyl)amino)methyl)-3- fluoroallyl)oxy)benzyl)thioureido)-1H-pyrrole-2-carboxylate (Int-21)

[0227] To a stirred solution of Int-21 (150 mg) in CH2Cl2 (2.0 mL) at rt was added benzoyl isothiocyanate (89 ^L). The resulting mixture was stirred at rt for 1 h. The reaction mixture was filtered and the filtrate was concentrated in vacuo. Purification was performed over silica gel (CombiFlash®), eluting with 10 – 100% ethyl acetate in hexane to afford Int-21 (110 mg, 54%). This material was used immediately in the next step. Procedure T: Preparation of tert-butyl (E)-(3-fluoro-2-((3-fluoro-5-((4-oxo-2-thioxo-2,3,4,5-tetrahydro- 1H-pyrrolo[3,2-d]pyrimidin-1-yl)methyl)phenoxy)methyl)allyl)carbamate (Int-22)

[0228] To a stirred solution of Int-21 (110 mg) in ethanol (2.0 mL) was added aqueous K2CO3 (0.3% w / w; 1.0 mL). The resulting mixture was heated to 110 °C in a microwave reactor, and stirring was continued at this temperature for 30 min. The reaction mixture then was partitioned between ethyl acetate and water. The organic layer was dried over Na2SO4 and then concentrated in vacuo. The crude material was purified over silica gel (CombiFlash®), eluting with 10 – 50% ethyl acetate in hexanes to afford Int-22 (25 mg, 11%) as a light grey solid.1H NMR (300 MHz, CDCl3) δ ppm: 11.11 (s, 1H), 9.94 (s, 1H), 7.22 (t, J = 3.0 Hz, 1H), 6.87 (s, 1H), 6.81 – 6.49 (m, 3H), 6.03 (t, J = 2.5 Hz, 1H), 5.68 (s, 2H), 4.41 (dd, J = 3.8, 1.2 Hz, 2H), 3.98 (d, J = 6.2 Hz, 2H), 1.43 (s, 9H).Procedure U: Preparation of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)-5-fluorobenzyl)-2-thioxo- 1,2,3,5-tetrahydro-4H-pyrrolo[3,2-d]pyrimidin-4-one hydrochloride (Compound 6)

[0229] To a stirred solution of Int-22 (23 mg) in MeOH (0.5 mL) was added HCl (2.0 M in diethylether; 2.0 mL). The resulting mixture was stirred at rt for 1 h during which time a white solid precipitated. The reaction mixture was then transferred to a vial and spun down in a centrifuge (4000 rpm, 4 min). The supernatant was carefully decanted and the solid “cake” was dried under vacuum to afford Compound 6 (13 mg, 65%) as a grey solid.1H NMR (300 MHz, Methanol-d4) δ ppm: 7.27 (d, J = 2.9 Hz, 1H), 7.21 (d, J = 80.6 Hz, 1H), 6.85 – 6.77 (m, 2H), 6.73 (dt, J = 10.6, 2.4 Hz, 1H), 6.13 (d, J = 2.9 Hz, 1H), 5.77 (s, 2H), 4.58 (d, J = 3.6 Hz, 2H), 3.79 (s, 2H). MS Found: 379 [M+H]+. EXAMPLE 7

[0230] Synthesis of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)-2-chlorobenzyl)-2-thioxo-1,2,3,5- tetrahydro-4H-pyrrolo[3,2-d]pyrimidin-4-one hydrochloride (Compound 7)

[0231] Preparation of tert-butyl (E)-(2-((2-chloro-3-formylphenoxy)methyl)-3-fluoroallyl)carbamate (Int-23)

[0232] Int-23 was prepared by modification of the protocol detailed in Procedure N from 2-chloro-3- hydroxybenzaldehyde and Int-6. In this instance, Cs2CO3was used in place of NaOtBu to afford Int-23 (563 mg, 100%).1H NMR (300 MHz, CDCl3) δ ppm: 10.54 (d, J = 0.8 Hz, 1H), 7.59 (dd, J = 7.8, 1.5 Hz, 1H), 7.36 (td, J = 8.0, 0.8 Hz, 1H), 7.20 (dd, J = 8.2, 1.5 Hz, 1H), 6.84 (d, J = 81.0 Hz, 1H), 4.58 (dd, J = 3.8, 1.2 Hz, 2H), 4.09 – 4.02 (m, 2H), 1.43 (s, 9H).Procedure V: Preparation of ethyl (E)-3-((3-((2-(((tert-butoxycarbonyl)amino)methyl)-3-fluoroallyl)oxy)- 2-chlorobenzyl)amino)-1H-pyrrole-2-carboxylate (Int-24)

[0233] To a stirred mixture of Int-23 (563 mg) and ethyl 3-amino-4-methyl-1H-pyrrole-2-carboxylate (140 mg) in MeOH (4.0 mL) at rt was added NaBH3CN (68.0 mg) followed by acetic acid (48 ^L). The resulting mixture was stirred at rt for 1 h. The reaction mixture was concentrated in vacuo. Attempts to purify this material by normal phase methods were unsuccessful. Crude Int-24 (360 mg) was obtained as a light yellow oil and carried forward to the next step.

[0234] Preparation of ethyl (E)-3-(3-benzoyl-1-(3-((2-(((tert-butoxycarbonyl)amino)methyl)-3- fluoroallyl)oxy)-2-chlorobenzyl)thioureido)-1H-pyrrole-2-carboxylate (Int-25)

[0235] Int-25 was prepared by modification of the protocol detailed in Procedure S from Int-24. Purification was performed over silica gel (Reveleris), eluting with 15 – 55% ethyl acetate in hexanes to afford Int-25 (260 mg) as a yellow solid.1H-NMR (300 MHz, CDCl3) complex, possibly due to hindered rotation; TLC Rf (0.5, 1:1 EtOAc : n-Hexane). Procedure W: Preparation of tert-butyl (E)-(2-((2-chloro-3-((4-oxo-2-thioxo-2,3,4,5-tetrahydro-1H- pyrrolo[3,2-d] 1- -

[0236] To a stirred mixture of Int-25 (260 mg) in EtOH (4.0 mL) at rt was added NaOtBu (114 mg) in one lot. The resulting mixture was stirred at rt for 2 d. HPLC analysis after this time indicated 25 - 30% of unchanged starting remained. The reaction mixture was concentrated in vacuo. The residue was partitioned between ethyl acetate (100 mL) and saturated aqueous NH4Cl (20 mL). The organic layer was washed withsaturated NaCl (20mL), dried over Na2SO4 and then concentrated in vacuo. The crude material was purified over silica gel (Reveleris®), eluting with 20 – 75% ethyl acetate in hexanes to afford Int-26 (120 mg, 60%) as a white solid.1H-NMR (300 MHz, CDCl3) δ ppm: 10.72 (s, 1H), 9.99 (s, 1H), 7.16 (t, J = 8.0 Hz, 1H), 7.01 (s, 1H), 6.91 (d, J = 8.2 Hz, 1H), 6.82 (d, J = 81.0 Hz, 1H), 6.49 (dd, J = 7.9, 1.2 Hz, 1H), 5.07 (s, 2H), 4.55 (d, J = 3.6 Hz, 2H), 4.07 (dd, J = 6.3, 2.2 Hz, 2H), 1.98 (s, 3H), 1.45 (s, 9H).

[0237] Preparation of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)-2-chlorobenzyl)-2-thioxo-1,2,3,5- tetrahydro-4H-pyrrolo[3,2-d]pyrimidin-4-one hydrochloride (Compound 7)

[0238] Compound 7 was prepared according to the experimental protocol detailed in Procedure U from Int-26; (96 mg, 91%) white solid.1H-NMR (300 MHz, Methanol-d4) δ ppm: 7.29 (d, J = 81.1 Hz, 1H), 7.25 (t, J = 8.0 Hz, 1H), 7.13 (d, J = 8.4 Hz, 1H), 7.08 (d, J = 0.9 Hz, 1H), 6.48 (dd, J = 7.7, 1.3 Hz, 1H), 5.95 (s, 2H), 4.75 (dd, J = 3.8, 1.0 Hz, 2H), 3.91 (d, J = 2.3 Hz, 2H), 1.95 (d, J = 0.8 Hz, 3H). MS Found: 393 [M+H]+. EXAMPLE 8

[0239] Synthesis of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)benzyl)-2-thioxo-1,2,3,5-tetrahydro- 4H-pyrrolo[3,2-d]pyrimidin-4-one hydrochloride (Compound 8)Procedure X: Preparation of - - - 1H-pyrrole (Int-27)

[0240] To a stirred solution of Int-4 (97 mg) and carbon tetrabromide (193 mg) in THF (3.0 mL) at rt was added triphenylphosphine (152 mg). The resulting mixture was stirred at rt for 1 h. The reaction mixture was concentrated in vacuo. The residue was purified over silica gel, eluting with 10% ethyl acetate in hexane to afford Int-27 (100 mg, 77%).1H-NMR (300 MHz, CDCl3) δ ppm: 6.86 (d, J = 81.3 Hz, 1H), 5.80 (s, 2H), 4.66 – 4.67 (m, 2H), 3.64 – 3.66 (m, 2H), 2.26 (s, 6H).

[0241] Preparation of (E)-3-((2-((2,5-dimethyl-1H-pyrrol-1-yl)methyl)-3-fluoroallyl)oxy)benzaldehyde (Int-28)

[0242] Int-28 was prepared according to the experimental protocol detailed in Procedure N from 3- hydroxybenzaldehyde and Int-27. Purification was performed over silica gel (CombiFlash®), eluting with 4 - 6% ethyl acetate in hexanes to afford Int-28 (1.10 g, 52%).1H NMR (400 MHz, CDCl3) δ ppm: 9.95 (s, 1H), 7.49-7.42 (m, 2H), 7.29-7.26 (m, 1H), 7.12-7.09 (m, 1H), 6.8 (d, J = 82 Hz, 1H), 5.74 (s, 2H), 4.7- 4.69 (m, 2H), 4.08 (d, J = 4 Hz, 2H), 2.13 (s, 6H). Procedure Y: Preparation of ethyl (E)-3-((3-((2-((2,5-dimethyl-1H-pyrrol-1-yl)methyl)-3- fluoroallyl)oxy)benzyl)amino)-1H-pyrrole-2-carboxylate (Int-29)

[0243] To a stirred solution of ethyl 3-amino-1H-pyrrole-2-carboxylate hydrochloride (0.50 g) and titanium isopropoxide (5.00 mL) at rt under N2 was added Int-28 (0.83 g). The resulting mixture was stirred at rt for 2 h under nitrogen atmosphere. After cooling to 0 °C, the reaction mixture was diluted with MeOH (10 mL). Sodium borohydride (0.50 g) was then added in portions and stirring was continued at rt for 5 h. The reaction mixture was then concentrated in vacuo. The residue obtained was purified over silica gel (Combiflash®, MPLC SilasepTM), eluting with 30 - 40% ethyl acetate in hexane to afford Int-29 (0.20 g, 18%).1H NMR (400 MHz, CDCl3) δ ppm: 7.2 (t, J = 4.8 Hz, 1H), 6.95 (d, J = 7.2 Hz, 1H), 6.84-6.83 (m, 2H), 6.7-6.68 (m, 2H), 5.73 (s, 2H), 5.64 (t, J = 2.8 Hz, 1H), 4.65 (s, 2H), 4.33-4.28 (m, 4H), 4.0 (d, J = 3.6 Hz, 2H), 2.15 (s, 6H), 1.35-1.31 (m, 3H); MS found: 426.3 (M+H)+. Procedure Z: Preparation of ethyl (E)-3-((3-((2-((2,5-dimethyl-1H-pyrrol-1-yl)methyl)-3- fluoroallyl)oxy) -

[0244] To a stirred solution of Int-29 (0.20 g) in CH2Cl2 at rt under N2 added benzoyl isothiocyanate (0.09 g). The resulting mixture was stirred at rt for 1 h. The reaction mixture was then concentrated in vacuo. The residue was dissolved in methanolic ammonia solution (7.0 M; 10 mL), transferred in a steel bomb, andthen heated at 80 °C for 4 h. The reaction mixture was again concentrated under reduced pressure. The residue obtained was purified over silica gel (CombiFlash®, MPLC SilasepTM) eluting with, 3 - 5% ethyl acetate in hexanes to afford Int-30 (95 mg, 48%) as an off-white solid.1H NMR (400 MHz, DMSO-d6) δ ppm: 12.35-12.25 (m, 2H), 7.31-7.28 (m, 2H), 7.2 (t, J = 7.6 Hz, 1H), 6.90-6.84 (m, 2H), 6.78-6.75 (m, 1H), 6.1 (s, 1H), 5.65 (s, 2H), 5.56 (s, 2H), 4.57 (s, 2H), 3.99 (d, J = 2.8 Hz, 2H), 2.03 (s, 6H); MS found: 439.3 (M+H)+. Procedure AA: Preparation of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)benzyl)-2-thioxo-1,2,3,5- tetrahydro-4H-pyrrolo[3,2-d]pyrimidin-4-one hydrochloride (Compound 8)

[0245] To a stirred solution of Int-30 (50.0 mg) in a 30 mL microwave vial at rt was added HCl (20% w / w in EtOH; 2.0 mL). The vial was capped and the mixture was heated in a microwave (Biotage) at 120oC for 45 min. The reaction mixture was concentrated in vacuo. These conditions were repeated on a further 3 batches (50 mg each). The residue from the combined batches were purified using prep- HPLC: [Waters Xbridge C18 (150 mm x 21.2 mm), A=0.05% HCl in water; B=MeCN Gradient, 10 - 50% B, Flow: 15.0 mL / min] and fractions were lyophilized to afford Compound 8 (27 mg, 60%) as an off white solid.1H- NMR (300 MHz, DMSO-d6) ^ ppm: 12.48 (s, 1H), 12.34 (s, 1H), 8.18 (s, 3H), 7.31 (d, J = 2.9 Hz, 1H), 7.30 (d, J = 82.1 Hz, 1H), 7.29 (d, J = 8.1 Hz, 1H), 6.98 – 6.93 (m, 2H), 6.93 – 6.87 (m, 1H), 6.13 (d, J = 2.9 Hz, 1H), 5.70 (s, 2H), 4.62 – 4.51 (m, 2H), 3.58 (d, J = 2.0 Hz, 2H); MS found 361.05 (M+H)+. COMPARATIVE EXAMPLE 9

[0246] Synthesis of (E)-3-(4-((2-(aminomethyl)-3-fluoroallyl)oxy)benzyl)-2-thioxo-1,2,3,7-tetrahydro- 6H-purin-6-one hydrochloride (Compound 9)Procedure AB: Preparation of tert-butyl (E)-(3-fluoro-2-((4-formylphenoxy)methyl)allyl)carbamate (Int- 31)

[0247] To a stirred solution of Int-5 (500 mg) and triethylamine (370 mg) in CH2Cl2(50.0 mL) at 0 °C was added MsCl (838 mg) by dropwise. The resulting mixture was then stirred at rt for 2 h. The reaction mixture was poured into water, and the product was extracted with ethyl acetate (100 mL x 3). The combined organics were washed with brine, dried over Na2SO4and concentrated in vacuo. The residue was taken up in DMSO (20.0 mL) and, after cooling to 0 °C, 4-hydroxybenzaldehyde (357 mg) was added followed by NaOtBu (309 mg). The resulting mixture was left to stir at rt overnight. The reaction mixture was poured into water, and the product was extracted with ethyl acetate (100 mL x 3). The combined organics were washed with brine, dried over anhydrous Na2SO4and concentrated in vacuo to afford Int-31 (850 mg). This material was used in the next step without further purification. MS Found: 310 [M+H]+. Procedure AC: Preparation of ethyl (E)-4-((4-((2-(((tert-butoxycarbonyl)amino)methyl)-3- fluoroallyl)oxy)benzyl)amino)- 5-carboxylate (Int-

[0248] To a stirred solution of Int-31 (754 mg) and ethyl 4-amino-1H-imidazole-5-carboxylate hydrochloride (0.48 g) in MeOH (3.0 mL) at rt was added NaBH3CN (0.15 g). The resulting mixture was stirred at rt for 2 h. The reaction mixture was then concentrated in vacuo. The residue was dissolved in ethyl acetate and washed with water. The aqueous layer was extracted with further ethyl acetate (100 mL x 2). The combined organic layers were dried over Na2SO4and then concentrated in vacuo. The crude product was purified over silica gel, eluting with 10% ethyl acetate in petroleum ether to afford crude Int-32 (1.10 g, 100%). This material was used in next step without further purification. MS Found: 420 [M+H]+.Procedure AD: Preparation of tert-butyl (E)-(3-fluoro-2-((4-((6-oxo-2-thioxo-1,2,6,7-tetrahydro-3H-purin- 3-yl)methyl)phenoxy)methyl)allyl)carbamate (Int-33)

[0249] To a stirred solution of benzoyl isothiocyanate (0.36 g) in CH2Cl2(1.0 mL) at 0 °C was added Int- 32 (545 mg). The resulting mixture was stirred at rt for 12 h. The solvent was removed in vacuo, and the residue was dissolved in aqueous NaOH (2.0% w / w; 27.0 mL). The resulting mixture was heated at 80 °C for 3 h. The pH of the reaction mixture was adjusted to neutral with 1M HCl. The resulting precipitate was collected by filtration, washed with water and purified by preparative-HPLC [Kinete EVO (Waters) C18 reverse-phase column (5 μm, 21.2 x 150mm) with a linear gradient of the binary solvent system 05 - 50% B, A:H2O (0.1% TFA), B: acetonitrile, with a flow rate of 15 mL / min and UV detection at 214 nm; GT 10min] to afford Int-33 (200 mg, 33%). MS Found: 461 [M+H]+. Procedure AE: Preparation of (E)-3-(4-((2-(aminomethyl)-3-fluoroallyl)oxy)benzyl)-2-thioxo-1,2,3,7- tetrahydro-6H-purin-6-one hydrochloride (Compound 9)

[0250] To a solution of Int-33 (200 mg) in ethyl acetate (20 mL) at 0 °C was added HCl (6.0 M in ethyl acetate; 15 mL). The resulting mixture was stirred at rt for 3 h. The solvent was concentrated in vacuo, and the residue was purified by preparative-HPLC [Gemininx C18 reverse-phase column (5 μm, 21.2 x 150mm) with a linear gradient of the binary solvent system 02 - 20% B, A: H2O (0.1% NH4HCO3), B: acetonitrile, with a flow rate of 15 mL / min and UV detection at 214 nm; GT 7 min] to afford Compound 9 (33.0 mg, 19%).1H-NMR (400 MHz, D2O) δ ppm: 8.47 (s, 1H), 7.38-7.36 (m, 2H), 7.14-6.94 (m, 1H), 6.89-6.87 (m, 2H), 4.54-4.53 (m, 2H), 4.38 (s, 2H), 3.80 (s, 3H); MS Found: 362 [M+H]+.COMPARATIVE EXAMPLE 10

[0251] Synthesis of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)-4-chlorobenzyl)-2-thioxo-1,2,3,5- tetrahydro-4H-pyrrolo[3,2-d]pyrimidin-4-one hydrochloride (Compound 10)

[0252] Preparation of tert-butyl (E)-(2-((2-chloro-5-formylphenoxy)methyl)-3-fluoroallyl)carbamate (Int-34)

[0253] Int-34 was prepared according to the experimental protocol detailed in Procedure AB from Int-5 and 4-chloro-3-hydroxybenzaldehyde; (430 mg). The crude material was used immediately in the next step without purification. MS found: 244 / 246 [M+H-Boc]+.

[0254] Preparation of ethyl (E)-3-((3-((2-(((tert-butoxycarbonyl)amino)methyl)-3-fluoroallyl)oxy)-4- chlorobenzyl)amino)-1H-pyrrole-2-carboxylate (Int-35)

[0255] Int-35 was prepared according to the experimental protocol outlined by Procedure AC from Int- 34 and ethyl 3-amino-1H-pyrrole-2-carboxylate. Purification was performed over silica gel (Reveleris®), eluting with 5 – 20% ethyl acetate in hexanes afforded Int-35 (150 mg, 25% over 2 steps).1H-NMR (300 MHz, CDCl3) δ ppm: 7.31 (d, J = 8.1 Hz, 1H), 6.96 (s, 1H), 6.92 (dd, J = 8.2, 1.8 Hz, 1H), 6.71 (d, J = 81.0 Hz, 1H), 6.69 (s, 2H), 4.52 – 4.42 (m, 2H), 5.16 (br. s, 1H), 4.40 – 4.23 (m, 4H), 4.01 (dd, J = 6.1, 2.4 Hz, 2H), 1.42 (s, 9H), 1.35 (t, J = 7.1 Hz, 3H).

[0256] Preparation of ethyl (E)-3-(3-benzoyl-1-(3-((2-(((tert-butoxycarbonyl)amino)methyl)-3- fluoroallyl)oxy)-4-chlorobenzyl)thioureido)-1H-pyrrole-2-carboxylate (Int-36)

[0257] Int-36 was prepared according to the experimental protocol detailed in Procedure S from Int-35. The crude material was purified over silica gel (Reveleris®), eluting with 10 – 100% ethyl acetate in hexanes to afford Int-36 (200 mg, 100%). MS Found: 645.2 [M+H]+.

[0258] Preparation of tert-butyl (E)-(2-((2-chloro-5-((4-oxo-2-thioxo-2,3,4,5-tetrahydro-1H-pyrrolo[3,2- d]pyrimidin-1-yl)methyl)phenoxy)methyl)-3-fluoroallyl)carbamate (Int-37)

[0259] Int-37 was prepared by modification of the experimental protocol detailed in Procedure T from Int-36. Aqueous K2CO3(2.0% w / w) was employed and the reaction mixture was heated at 110 °C, under microwave irradiation, for 20 min. The crude material was purified by reverse-phase chromatography (Reveleris®), eluting with 40 – 90% CH3CN in water to afford Int-37 (41 mg, 29%).1H-NMR (300 MHz, CDCl3) δ ppm: 7.32 (d, J = 8.1 Hz, 1H), 7.18 (d, J = 2.9 Hz, 1H), 7.12 (s, 1H), 6.92 (dd, J = 8.1, 2.0 Hz, 1H), 6.75 (d, J = 84.0 Hz, 1H), 6.05 (d, J = 3.0 Hz, 1H), 5.68 (s, 2H), 5.17 (br. s, 1H), 4.50 (dd, J = 3.8, 1.2 Hz, 2H), 4.06 – 3.96 (m, 2H), 1.43 (s, 9H).

[0260] Preparation of (E)-1-(3-((2-(aminomethyl)-3-fluoroallyl)oxy)-4-chlorobenzyl)-2-thioxo-1,2,3,5- tetrahydro-4H-

[0261] Compound 10 was prepared according to the experimental protocol outlined in Procedure U from Int-37; (30 mg, 84%).1H NMR (300 MHz, Methanol-d4) δ ppm: 7.39 (d, J = 8.1 Hz, 1H), 7.30 – 7.21 (m,2H), 7.23 (d, J = 81.1 Hz, 1H), 7.04 (dd, J = 8.2, 1.9 Hz, 1H), 6.17 (d, J = 3.0 Hz, 1H), 5.79 (s, 2H), 4.66 (dd, J = 3.6, 1.0 Hz, 2H), 3.84 (d, J = 1.8 Hz, 2H); MS Found: 395.1 [M+H]+. EXAMPLE 11

[0262] Method to determine the ability of compounds of Formula I to inhibit human recombinant SSAO / VAP-1

[0263] Human recombinant SSAO / VAP-1 amine oxidase activity was determined using the coupled colorimetric method as described for monoamine oxidase, copper-containing amine oxidases and related enzymes (Holt A. and Palcic M., A peroxidase-coupled continuous absorbance plate-reader assay for flavin monoamine oxidases, copper-containing amine oxidases and related enzymes. Nat Protoc 2006, 1, 2498- 2505). Briefly, a cloned cDNA template corresponding to residues 34-763 of human SSAO / VAP-1, and incorporating a mouse Ig kappa (κ) signal sequence, N-terminal flag epitope tag and tobacco etch virus (TEV) cleavage site, was assembled in a mammalian expression vector (pLO-CMV) by Geneart AG. This vector containing human SSAO / VAP-1 residues was transfected into CHO-K1 glycosylation mutant cell line, Lec 8. A clone stably expressing human SSAO / VAP-1 was isolated and cultured in large scale. Active human SSAO / VAP-1 was purified and recovered using immunoaffinity chromatography. This was used as the source for SSAO / VAP-1 activity. A high-throughput colorimetric assay was developed using either 96 or 384 well format. Briefly, in a standard 96 well plate assay 50 µL of purified human SSAO / VAP-1 (0.25 µg / mL) in 0.1 M sodium phosphate buffer (pH 7.4) was added into each well. Test compounds were dissolved in DMSO and tested in a Concentration Response Curve (CRC) with 4-11 data points, typically in the micromolar or nanomolar range after incubation with human SSAO / VAP-1 for 30 min at 37 ^C. After 30 min incubation, 50 µL of the reaction mixture containing 600 µM benzylamine (Sigma Aldrich), 120 µM Amplex Red (Sigma Aldrich) and 1.5 U / mL horseradish peroxidase (Sigma Aldrich) prepared in 0.1 M sodium phosphate buffer (pH 7.4) were added to the corresponding well. The fluorescence unit (RFU) was read every 2.5 min for 30 min at 37^C excitation 565nm and emission 590 (Optima; BMG labtech). The slope of the kinetics for each well was calculated using MARS data analysis software (BMG labtech) and this value was used to deduce the IC50 value (Dotmatics). The ability of the compounds of Formula I to inhibit SSAO / VAP-1 is shown in Table 2. EXAMPLE 12

[0264] Method to determine the ability of compounds of Formula I to inhibit human native MPO – Amplex Red method (AR)

[0265] Human MPO activity was determined using the colorimetric method as described (Ward et al., Biochemistry 2013, 52, 9187-9201). Purified human MPO from polymorphonuclear leukocytes wasobtained from Calbiochem®(cat 475911-100µg) and used as the source for MPO activity. A high- throughput colorimetric assay was used in either 96 or 384 well format. Briefly, in a standard 384 well plate assay 25 µL of purified human MPO (500 pM) in Assay Buffer (50 mM sodium phosphate, 150 mM sodium chloride, 1 mM DPTA and 2% DMSO, pH 7.4) was added into each well. Test compounds were dissolved in DMSO and tested in a Concentration Response Curve (CRC) with 4-11 data points, typically in the micromolar or nanomolar range after incubation with human MPO for 30 min at 37 ^C. After 30 min incubation, 25 µL of the reaction mixture containing 60 µM Amplex Red (Sigma Aldrich) and 100 µM H2O2 (Sigma Aldrich) prepared in Assay Buffer were added to the corresponding well. The fluorescence unit (RFU) was read every 2.5 min for 30 min at 37 ^C excitation 565 nm and emission 590 nm (Optima; BMG labtech). The slope of the kinetics for each well was calculated using MARS data analysis software (BMG labtech) and this value was used to deduce the IC50 value (Dotmatics). The ability of the compounds of Formula I to inhibit MPO is shown in Table 2. EXAMPLE 13

[0266] Method to determine the ability of compounds of Formula I to inhibit human recombinant monoamine oxidase A (MAO-A)

[0267] MAO-A inhibitors have the potential to induce hypertensive crisis when dietary tyramine is ingested and therefore come with certain dietary limitations. Thus, for medicinal applications a high selectivity over MAO-A is required.

[0268] The selectivity of the compounds of this invention was tested by determining their ability to inhibit MAO-A activity in vitro, using recombinant human MAO-A (0.003 mg / mL; Sigma Aldrich). The assay was performed in a similar way as for human SSAO / VAP-1 (EXAMPLE 11) except the substrate tyramine was used at 100 µM in place of benzylamine.

[0269] Compounds of Formula I inhibit MAO-A with an IC50 > 1 ^M. EXAMPLE 14

[0270] Method to determine the ability of compounds of the invention to inhibit lysyl oxidase from different sources

[0271] Lysyl oxidase (LOX) is an extracellular copper dependent enzyme which oxidizes peptidyl lysine and hydroxylysine residues in collagen and lysine residues in elastin to produce peptidyl alpha- aminoadipic-delta-semialdehydes. This catalytic reaction can be irreversibly inhibited by β- aminopropionitrile (BAPN) that binds to the active site of LOX. LOX can be extracted from animal aorta or prepared from human cell cultures.

[0272] Under normal physiological conditions, LOX is ubiquitously expressed in most human tissues at high levels and is essential in the development of the healthy bone, respiratory system, and cardiovascular function. In the context of drug development, a high selectivity over LOX is required.

[0273] The inhibitory effects of the compounds of the present invention were tested against the given LOX preparation using a method based on the detection of hydrogen peroxide with an Amplex Red oxidation assay (Zhou. et al. Anal. Biochem. 1997, 253, 162-168). The assay was developed using either 384 or 96 well format. Briefly, in a standard black, clear bottom 384 well plate assay 25 μL of a dilution of LOX enzyme in 1.2 M urea, 50 mM sodium borate buffer (pH 8.2) were added into each well in the presence of 1 μM mofegiline and 0.5 mM pargyline (to inhibit SSAO / VAP-1 / MAO-B and MAO-A, respectively). Test compounds were dissolved in DMSO and tested in a Concentration Response Curve (CRC) with 11 data points, typically in the micromolar or nanomolar range after incubation with the enzyme for 30 min at 37 °C.25 μL of a reaction mixture containing twice the KMconcentration of putrescine (Sigma Aldrich, 20 mM), 120 μM Amplex Red (Sigma Aldrich) and 1.5 U / mL horseradish peroxidase (Sigma Aldrich) prepared in 1.2 M urea, 50 mM sodium borate buffer (pH 8.2) were then added to the corresponding wells. The above volumes were doubled in the case of 96 wells plate. The fluorescence (RFU) was read every 2.5 min for 30 min at a range of temperatures from 37 °C, excitation 565nm and emission 590 (Optima; BMG labtech). The slope of the kinetics for each well was calculated using MARS data analysis software (BMG labtech) and this value was used to deduce the IC50value (Dotmatics).

[0274] Compounds of Formula I inhibit the LOX protein with an IC50 > 1 ^M. EXAMPLE 15

[0275] Method to determine the ability of compounds of Formula I to inhibit human recombinant thyroid peroxidase (TPO)

[0276] Thyroid peroxidase (TPO), is a peroxidase closely related to MPO that is involved in the production of key thyroid hormone proteins. In the context of drug development, a high selectivity over TPO is required.

[0277] The selectivity of the compounds of this invention was tested by determining their ability to inhibit TPO activity in vitro, using recombinant human TPO (Abacus dx). Compounds were diluted to desired concentrations in PBS (10 mM Na2HPO4, 140 mM NaCl), pH 7.4, supplemented with ascorbic acid 100 μM), DTPA (100 μM), and NaI (50 μM). TPO (200 nM) was added to each tube and mixture incubated for 5 mins at rt before initiating reaction. Reaction mixtures were transferred to UV-transparent 96-well assay plates (UV-Star®, Sigma-Aldrich) and reactions were initiated by addition of H2O2 (25 μM), and rapidly mixed. Final reaction volume was 210 μL. Absorbance at 266 nm was immediately measured witha plate reader (Spectramax M3; Molecular Devices). Ascorbate oxidation was continuously monitored every 20 seconds for 6 min. Ascorbate oxidation rate was expressed as a percentage of control with reaction rate calculated from 0-200 sec of the reaction.

[0278] Compounds of Formula I inhibit TPO with an IC50 > 1 ^M. Table 2. A <0.1; 0.1< B <1; 1< C <10; D >10.*Table 2 Footnote: 9 and 10 are Comparative examples, that are structurally similar but not part of the current invention. EXAMPLE 16

[0100] Pharmacokinetic parameters of Compound 1

[0101] Pharmacokinetic (PK) studies were carried out at either GVK Bio, Hyderabad (Wistar rat), or Sundia MediTech Co. Ltd., Shanghai (beagle dog, C57 mouse). Animals were administered Compound 1 by either intravenous injection or by oral gavage at the specified doses. At various times between 15 minutes and 8 hours, blood samples were collected. Plasma was analyzed by high performance liquid chromatography–tandem mass spectrometry. The basic PK parameters thus obtained are outlined in Table 3.Table 3. Pharmacokinetic profile of Compound 1 in rats, dogs and mice Species rat (Wistar) dog (Beagle) mouse (C57) I O C A C T FEXAMPLE 17

[0279] Method to determine the long lasting inhibition of target enzymes of compounds of Formula I

[0280] Long lasting inhibition of the target enzymes may be an advantage, leading to more extended effects, even after the concentration of a pharmacologically active substance in circulation has fallen below efficacious levels. This example investigates the ability of compounds to continue to inhibit enzyme activity after rapid dilution, based on the protocol outlined by Ward et. al. Biochemistry, 2013, 52, 9187−9201.

[0281] MPO was pre-incubated in the presence of 2 μM H2O2with Compound 1 at concentration approximately 30-fold higher than the IC50value, or DMSO as a control for 15 min at room temperature. The reaction mixture was then rapidly diluted 300-fold into the assay buffer containing of 50 mM sodium phosphate buffer (pH 7.4), 1 mM diethylenetriaminepentaacetic acid, 2 µM H2O2and 30 μM Amplex Red. The reaction was well mixed and 100 μL transferred to a 96-well assay plate. The target percentage activity was measured as a function of time after the dilution event.

[0282] For SSAO / VAP-1 the measurement of the residence time was based on the detection of hydrogen peroxide with an Amplex Red oxidation assay, and as described in EXAMPLE 11 in a 96 well format. The protein was incubated with Compound 1 at 30 x IC50for 60 minutes at 37 °C in SnapStrip PCR vials. After the incubation, a 50-fold dilution was carried out by adding the assay buffer 0.1 M sodium phosphate buffer (pH 7.4), immediately followed by a 1:2 dilution with an equal volume of reaction mixture (a mixture of 600 µM benzylamine (Sigma Aldrich), 120 µM Amplex Red (Sigma Aldrich) and 1.5 U / mL horseradish peroxidase (Sigma Aldrich) prepared in 0.1 M sodium phosphate buffer (pH 7.4)) and subjected to measurement as described in the enzymatic assay methodology. The target percentage activity was measured as a function of time after the dilution event.

[0283] Compound 1 displayed behaviour consistent with long-lasting inhibition of both of the target enzymes.EXAMPLE 18

[0284] Efficacy of Compound 1 in a lipopolysaccharide (LPS)-induced lung inflammation model

[0285] Female Balb / cJUniB mice received a single oral gavage of the vehicle or Compound 1 at 30 mg / kg dissolved in vehicle (0.5% carboxymethylcellulose and 0.25% TWEEN20®in PBS). After one hour, mice were anesthetized via subcutaneous injection with a mixture of ketamine 10 mg / kg and xylazine 15 mg / kg (both Syntec) diluted in sterile saline, then intranasally administered with LPS, 25 µg per animal, in a volume of 30 µL of sterile saline. Six hours after LPS installation mice received a lethal dose of ketamine / xylazine

[0286] Mice were cannulated in the trachea and 1 mL of PBS was gently flushed three times in and out of the lung, followed by a further 1 mL. As a result, a total volume of ~1,8 mL of brochoalveolar lavage fluid (BALF) was obtained (~90% recovery of the total flushed volume).

[0287] BALF was centrifuged at 4 ºC, 1500 rpm for 8 minutes and the resultant pellet was resuspended in 100 µL of saline solution with 3% of BSA (bovine serum albumin). Cells were added to cytospin slide with 3% BSA and centrifuged at 450 rpm for 5 minutes (Shandon III). Slides were stained for May- Grunwald-Giemsa and the differential counts were performed. Cells were differentiated into mononuclear cells, neutrophils and eosinophils based on morphology.

[0288] The anti-inflammatory effects of Compound 1 in the lung are shown in Figure 1. Compound 1 significantly reduced total inflammatory cell and neutrophil influx measured in the BALF. Data were analyzed by student’s t test. *p < 0.05, **p < 0.01, ***p < 0.001 and ****p < 0.0001 compared to non- treated mice (bottom value) or vehicle treated animals (top value). EXAMPLE 19

[0289] Efficacy of Compound 1 in a thioglycolate broth (TGB) - induced peritonitis model

[0290] C57BL / 6 mice received a single oral gavage of the vehicle or Compound 1 at 60 mg / kg dissolved in vehicle (0.5% hydroxypropyl methyl cellulose (HPMC) (Sigma-Aldrich), with 0.1% (w / v) Tween 80 in water). Mice undergoing lavage at 12 hours after inhibitor treatment, received a repeat dose of drug by oral gavage 6 hours after TGB treatment. After one hour, mice received a 1 mL intra-peritoneal injection of 4% thioglycolate broth (TGB) (prepared by dissolving 4% (w / v) thioglycolate medium (Becton Dickinson) in distilled water with heat) to induce leukocyte recruitment into the peritoneum.

[0291] At 6 or 12 hours after administration of Compound 1, mice were euthanized by CO2asphyxiation, and the peritoneal cavity was lavaged with 1 mL of ice-cold PBS solution. Total cell number in exudateswas determined by haemocytometer. Cell counts of leukocyte subtypes (neutrophils and monocytes) were determined using differential staining (Diff Quik).

[0292] The anti-inflammatory effects of Compound 1 in the peritoneum are shown in Figure 2. Compound 1 significantly reduced total inflammatory cell and neutrophil influx measured in the peritoneal lavage. Data were analyzed by student’s t test. *p < 0.05 and **p < 0.005 compared to TGB-vehicle treated mice. EXAMPLE 20

[0293] Method to determine the ability of compounds of Formula I to inhibit mouse peritoneal MPO.

[0294] C57BL / 6 mice initially received a 1 mL intra-peritoneal injection of 4% thioglycollate broth (TGB), to induce leukocyte recruitment into the peritoneum. The broth was prepared by adding 4% (w / v) thioglycollate medium (Becton Dickinson) to distilled water, and the solution was brought to boil with constant mixing. After 20 hours, mice received a single dose (via oral gavage) of Compound 1 at 10, 30, or 60 mg / kg of body weight dissolved in vehicle (0.5% Hydroxypropyl methyl cellulose (HPMC) (Sigma- Aldrich), with 0.1% (w / v) Tween 80 in water). One hour after oral gavage, leukocytes were activated by intra-peritoneal injection with 0.5 mL of Zymosan A (10 mg / mL) (from Saccharomyces cerevisiae, Sigma- Aldrich) in 0.9% saline (Baxter). Four hours after Zymosan A treatment, mice were euthanased by CO2 asphyxiation, and the peritoneal cavity was lavaged with 1 mL of ice-cold PBS. Peritoneal exudate cells were pelleted by centrifugation (4,500 rpm, 5 min at 4°C). MPO activity in peritoneal exudates was determined using a bioluminescence assay, measuring MPO Activity on a Polymer Surface (MAPS) (as described by Goiffon, Martinez et al.2015). Lavage samples were diluted with MAPS loading buffer (130 mM NaCl, 6.28 mM Na2HPO4, 18.7 mM NaH2PO4, and 50 ppm v / v Tween 20) to a total volume of 500 μL. Prior to bioluminescence assay, the concentration of MPO in lavage fluid was determined with an ELISA kit (Hycult) to allow for MPO normalisation to a positive control. Samples were thoroughly mixed then 100 μL / well loaded into a black-walled, tissue culture treated 96-well microtiter plate (Corning). The microtiter plate was covered and shaken at 1,000 rpm for 30 min at RT, to facilitate protein binding to well surfaces. Subsequently, each well was aspirated and washed three times with MAPS loading buffer: twice with 150 μL, then once with 300 μL. Each well then received 75 μL of freshly prepared MAPS imaging solution (3.1 mM citric acid, 21.9 mM trisodium citrate, 20 mM NaBr, 200 mM (NH4)2SO4, and 100 ppm v / v Tween 20) supplemented with 50 μM L-012 (Wako), and 100 μM H2O2immediately before use. The plate was briefly shaken at 1,000 rpm and read immediately. Bioluminescence was continuously monitored every 20 seconds for 16 min, using an EnSpire® Multimode Plate Reader (PerkinElmer). Area Under Curve (AUC), of Relative Luminescence Units (RLU) over 16 min, was calculated with data expressed as a percentage of positive control.

[0295] The in vivo inhibition of peritoneal activated MPO by Compound 1 is shown in Figure 3. Stimulation of the inflammatory infiltrate by Zymosan A generates activated MPO that is then inhibited by Compound 1 in a dose-dependent manner. EXAMPLE 21

[0296] Efficacy of Compound 1 in the carrageenan air pouch model.

[0297] On day 0, BALB / c mice were shaved at the nape of the neck, anesthetized and 6 mL of sterile air (0.2 μm, Sartorius, Cat.16532) was injected subcutaneously using a 23G x 1½ inch needle fixed to a 6 mL syringe. The injection site was sealed with flexible collodion (Macron, Cat.4580-02). On day 3, a further 3 mL of sterile air was injected into the neck. 3% λ-carrageenan was prepared by dissolving 3 g λ- carrageenan (FLUKA, Cat. 22049, lot 1408463) in 80 mL hot de-ionized water. Once dissolved, the solution was cooled to room temperature and the volume made up to 100 mL with de-ionized water. On day six, Compound 1 in 30% (2-hydroxypropyl)-β-cyclodextrin (vehicle) or vehicle only were administered by oral gavage to the mice.30 minutes after administration, mice were injected with 1 mL 3% λ-carrageenan, using a 23G x 2-inch needle fitted to a 1 mL syringe, directly into the air pouch. Sham animals were administered 1 mL de-ionized water instead of λ-carrageenan solution. Eight hours after carrageenan injection, the mice were re-dosed with either Compound 1 or vehicle.

[0298] Twelve hours after carrageenan / de-ionized water injection, the mice were anesthetized and 3 mL 10 U / mL heparinized saline was injected into the air pouch. The air pouch was gently massaged, the contents immediately removed using an 18G x 1-inch needle fitted to a 5 mL syringe, and the exudate volume recorded. An aliquot of the exudate taken for total white blood cell counting; a second aliquot of the exudate was taken for differential white blood cell counts.

[0299] The anti-inflammatory effects of Compound 1 are shown in Figure 4. Carrageenan injection caused an increase in inflammation, which was reduced by application of Compound 1, as seen by a decrease in both exudate volume and total white blood cell (WBC) count in the air pouch. *p<0.05 compared to carrageenan-treated mice. EXAMPLE 22

[0300] Method to determine the ability of compounds of Formula I to inhibit mouse adipose tissue SSAO / VAP-1

[0301] BALB / c mice from the study outlined in EXAMPLE 21 were sacrificed at various time points and gonadal fat tissue collected.

[0302] Adipose tissue was homogenized in ice-cold HES buffer (20 mM HEPES, 1 mM EDTA, sucrose 250 mM, proteases and phosphatases inhibitor, pH 7.4). Homogenate was then centrifuged at 2000 rpm for 5 minutes at 4oC and the resultant supernatant portion (minus the lipid top layer) used for assay. SSAO / VAP-1 activity was determined by using a non-specific MAO inhibitor (e.g. pargyline) to establish the high signal and a mixture of non-specific MAO inhibitor and an SSAO / VAP-1 inhibitor (e.g. semicarbazide) to determine the low signal.

[0303] Activity of the treated samples was analyzed using the fluorometric method described in EXAMPLE 11 for SSAO / VAP-1 activity.

[0304] The in vivo inhibition of SSAO / VAP-1 by Compound 1 is shown in Figure 5. Animals subjected to the Carrageenan Air pouch model were sacrificed on day 6 of the experiment at either 4 h or 12 h post initial administration of Compound 1. EXAMPLE 23

[0305] Efficacy of Compound 1 in a rat model of myocardial infarction. [HRI]

[0306] Cardiac I / R (ischemia reperfusion) injury was induced in male Wistar rats by a transient 30 min ligation of the left anterior descending coronary artery and animals were dosed by oral gavage twice a day for the following four weeks.

[0307] At the end of the experiment, animals were euthanized and the hearts removed and sliced. The heart was fixed with 10% formalin. Fibrosis was assessed in the non‐infarct area using both Masson’s Trichrome and Picrosirius Red staining. Fibrotic area and whole non‐infarct area were measured using computerized planimetry (Image J). The fibrotic area was presented as a percentage of the whole non‐ infarct area. Three random fields per heart were counted, averaged and a total of 30‐45 fields per group were measured.

[0308] The anti-fibrotic effects of Compound 1 are shown in Figure 6. In a rat model of myocardial infarction, induced by ligation of the left descending coronary artery, Compound 1 was administered twice daily at 60 mg / kg for 4 weeks following surgery, affording a significant reduction in fibrosis. *p<0.05 and **p<0.01 vs control. EXAMPLE 24

[0309] Efficacy of Compound 1 in the unilateral ureteral obstruction (UUO) acute kidney injury model.

[0310] UUO was performed at Urosphere SAS, Toulouse, France, following the procedure described by Schanstra et al., J. Clin. Invest.2002, 110, 371-379. Briefly, mice were anesthetized with a mix of oxygen-isoflurane (0.2 L / min and 1.5-2% isoflurane) and then maintained at 37 °C using a thermo-regulated system during surgery. A cutaneous incision was performed on the left flank, followed by a muscular incision. The left ureter was exposed, and ligated (6 / 0 silk). Then, the flank incision was sutured (muscle wall and the cutaneous wall) and disinfected with antimicrobial agent (Vetadine®, Centravet, France). After surgery, the animals were returned to their cages.

[0311] Compound 1 was dissolved at a concentration of 13.21 mg / mL in 0.6% methylcellulose solution (Sigma –Aldrich). Captopril was dissolved at a concentration of 10 mg / mL in deionized water. Compound 1 was administered twice a day at 60 mg / kg by oral gavage. Captopril was administered via the drinking water to a concentration of ~32 mg / kg / day. Dosing was continued from day 1 after surgery until day 13.

[0312] At the end of the protocol (14 days after UUO) animals were sacrificed by cervical dislocation. After sacrifice, the right and left kidneys were removed, cleaned from all around tissues, rinsed in physiological saline and blotted on a paper. Each kidney (right and left) was weighed and the length and thickness measured. Left and right kidneys were placed in separated embedding cassettes into formalin 10% solution at room temperature for 24 h. The fixed kidneys were then transferred in 70% ethanol until histological preparation.

[0313] Kidneys were embedded in paraffin blocks and two sections (3 to 5 μm) were made per block. Slides were stained with Sirius Red for histological evaluation of fibrosis (interstitial collagen accumulation) and digitalized at X20 magnitude using Nanozoomer 2.0 HT from Hamamatsu (Hamamatsu, Japan). Kidney fibrosis was quantified using Image J software. Non-renal tissue (vascular, perivascular and adipose tissue) was not included in the quantification of fibrosis (red stained surface).

[0314] The anti-fibrotic effects of Compound 1 are shown in Figure 7. In a 14-day model of acute kidney injury (AKI), induced by unilateral ureteral obstruction (UUO), Compound 1 dosed twice a day from day 1 significantly reduced fibrosis in the obstructed kidney, as measured by Picrosirius Red staining. *p<0.05 vs UUO / vehicle (one-way Anova, Dunnett’s post-test) $ p<0.05, $$ p<0.01 vs UUO / vehicle (unpaired student t-test) ### p<0.001 left kidney vs right kidney of UUO / vehicle (unpaired student t-test with Welch’s correction). EXAMPLE 25

[0315] Assessment of the metabolic stability profile of Compound 1 using standardized commercially available methods (available through contract research organizations such as Eurofins Panlabs Inc, (https: / / www.eurofinsdiscoveryservices.com / services / in-vitro-assays / ) or Evotec SE, (https: / / www.evotec.com / en / execute / integrated-pre-clinical-development / adme)).

[0316] The potential for metabolism in vivo was evaluated in vitro using the following assays:

[0317] Liver microsomal stability (human, rat, dog; in presence of NADPH)

[0318] Hepatocyte stability (human, rat, dog)

[0319] Plasma stability (human, rat, dog)

[0320] Compound 1 was found to have favourable metabolic stability. EXAMPLE 26

[0321] Assessment of the off-target profile of Compound 1 using standardized commercially available methods (available through contract research organizations such as Eurofins Panlabs Inc, (https: / / www.eurofinsdiscoveryservices.com / services / in-vitro-assays / ) or Evotec SE, (https: / / www.evotec.com / en / execute / integrated-pre-clinical-development / adme)).

[0322] The potential for off-target promiscuity in vivo was evaluated in vitro using the following assays:

[0323] Inhibition of cytochrome P450 activity (30 ^M single concentration; 1A2, 2B6, 2C8, 2C9, 2C19, 2D6, 3A4)

[0324] Eurofins Discovery Lead Profiling Screen®panel of 68 enzymes, receptors, ion channels and transporters (percentage inhibition at a single 10 ^M concentration).

[0325] Eurofins Transporter Protein Screen panel of 13 common transporters (percentage inhibition at a single 30 ^M concentration).

[0326] Compound 1 was found to have a favorable off-target profile. EXAMPLE 27

[0327] Assessment of the mutagenicity potential of Compound 1 in a miniaturized bacterial reverse mutagenicity screening assay.

[0328] Compound 1 was tested for mutagenic potential in a range-finding mutagenicity assay at single plates per dose, with and without metabolic activation (±S9). The assay was conducted using the following five bacterial strains that are prescribed in the test guideline for the bacterial reverse mutation assay (OECD, 1997): Salmonella typhimurium TA98, TA100, TA97a, and TA1535, and Escherichia coli WP2 uvrA pKM101. Metabolic activation was provided by a 10% phenobarbital / benzoflavone-induced rat liver S9 mix. A high concentration stock solution was prepared from the neat test article at 50 mg / mL in sterilewater. The remaining dose formulations were prepared as serial dilutions from the high-concentration stock solution. Nominal formulation concentrations were 50, 25, 12.5, 5, 2.5, 1.25, 0.5, and 0.2 mg / mL and nominal test doses were 5000, 2500, 1250, 500, 250, 125, 50, and 20 μg / plate. The assay was performed using the pre-incubation method in which the assay tubes were incubated at 37 ± 1°C for 20 ± 1 minutes before plating onto minimal agar. Test plates were incubated at 37 ± 1°C for 48 ± 2 hours and then counted using the Sorcerer / Ames Study Manager system from Perceptive Instruments.

[0329] Compound 1 did not induce a response indicative of mutagenic potential in this assay. EXAMPLE 28

[0330] Assessment of the potential of Compound 1 to induce phospholipidosis in a cell-based fluorescence assay.

[0331] Phospholipidosis is the result of undesired interference with normal cellular functions that may be associated with drug candidates, particularly such that combine a basic, positively ionisable group with lipophilic features.

[0332] Compound 1 was tested in vitro for the potential to induce phospholipidosis according to the method outlined by Fujimura et. al. (Experimental and Toxicologic Pathology, 2007, 58, 375–382) at concentrations up to 100 ^M.

[0333] Compound 1 had no effect on the cellular phenotype nor cell viability in this assay.

Claims

CLAIMS 1. A compound of formula I:or a pharmaceutically acceptable salt thereof; wherein: R1and R2are independently selected from the group consisting of hydrogen and fluorine; provided that R1and R2are not hydrogen at the same time; X is selected from the group consisting of N, CH and C-CH3; Y is selected from the group consisting of hydrogen, fluorine and chlorine; and Z is hydrogen or fluorine.

2. A compound according to Claim 1, wherein R1is hydrogen and R2is fluorine 3. A compound according to Claim 1 or 2, wherein X is N.

4. A compound according to Claim 1 or 2, wherein X is CH.

5. A compound according to Claim 1 or 2, wherein X is C-CH3 6. A compound according to Claim 1 of Formula Ia:or a pharmaceutically acceptable salt thereof; wherein: X is selected from the group consisting of N, CH and C-CH3; Y is selected from the group consisting of hydrogen, fluorine and chlorine; andZ is hydrogen or fluorine.

7. A compound according to Claim 1 selected from the group consisting of: C m nd - -- -or a pharmaceutically acceptable salt thereof.

8. A pharmaceutical composition comprising a compound according to any one of Claims 1 to 7, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient, carrier or diluent.

9. A method of inhibiting the amine oxidase activity of SSAO / VAP-1 and the peroxidase activity of MPO in a subject in need thereof, comprising administering to the subject an effective amount of a compound according to any one of Claims 1 to 7, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to Claim 8.

10. A method of treating or preventing a disease by inhibiting the activity of the SSAO / VAP-1 protein and the MPO protein, comprising administering to a subject in need thereof a therapeutically effective amount of a compound according to any one of Claims 1 to 7, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to Claim 8.

11. The method according to Claim 10, wherein the disease is associated with inflammation or fibrosis.

12. The method according to Claim 10, wherein the disease is associated with a reduction in kidney function, and the cause of this reduction in kidney function is selected from the group consisting of acute tubular necrosis (ATN), severe or sudden dehydration, toxic kidney injury from poisons or certain medications, the result of a severe bacterial or viral infection, such as COVID-19, urinary tract obstruction and autoimmune kidney diseases, such as acute nephritic syndrome and interstitial nephritis.

13. The method according to Claim 10, wherein the disease is chronic kidney disease, and the cause of the chronic kidney disease is selected from the group consisting of obesity, type 1 or type 2 diabetes, including diabetic nephropathy, high blood pressure, glomerulonephritis, interstitial nephritis, polycystic kidney disease, prolonged obstruction of the urinary tract from conditions such as cancer, heart failure, heart failure with preserved ejection fraction, heart failure with reduced ejection fraction, vasculitis (including anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis), enlarged prostate or kidney stones, vesicoureteral reflux and pyelonephritis.

14. The method according to Claim 10, wherein the disease is a respiratory disease selected from the group consisting of asthma, neutrophilic asthma, chronic obstructive pulmonary disease, lung fibrosis, including idiopathic pulmonary fibrosis, bronchiectasis, interstitial lung disease or resulting from severe bacterial or viral infection, mesothelioma, asbestosis, silicosis, effects of severe bacterial or viral infection in the lung such as acute respiratory distress syndrome or acute lung injury as a result of a cytokine storm, ventilator associated pneumonia, and pulmonary complications arising as a result thereof.

15. The method according to Claim 10, wherein the disease is a cardiovascular disease selected from the group consisting of hypertension, abnormal heart rhythms, or arrhythmias, atherosclerosis, aorta disease and Marfan syndrome, congenital heart disease, cardiomyopathy, coronary artery disease, deep vein thrombosis and pulmonary embolism, dyslipidemias, heart attack, heart failure, heart failure with preserved ejection fraction, heart failure with reduced ejection fraction, cardiac fibrosis, including viral or bacterial induced inflammation and fibrosis, pericardial disease, peripheral vascular disease, rheumatic heart disease, stroke and vasculitis (including anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis).

16. The method according to Claim 10, wherein the disease is a liver disease selected from the group consisting of liver fibrosis, including alcoholic liver disease, fatty liver disease, non-alcoholic steatohepatitis (NASH) / metabolic dysfunction-associated steatohepatitis (MASH), acute and chronic hepatitis, biliary cirrhosis, primary sclerosing cholangitis, liver autoimmune diseases, hepatocellular carcinoma and toxic liver injury.

17. The method according to Claim 10, wherein the disease is an eye disease selected from the group consisting of uveitis, diabetic macular oedema, glomerulosclerosis, diabetic retinopathy, age-related macular degeneration, ocular scarring and conjunctival inflammatory disease.

18. The method according to Claim 10, wherein the disease is selected from the group consisting of a fibrotic disease and a neurological disease.

19. Use of a compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for treating or preventing a disease by inhibiting the activity of the SSAO / VAP-1 protein and the MPO protein.

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