BR112018073700B1 - TRPV4 ANTAGONIST COMPOUND, PHARMACEUTICAL COMPOSITION COMPRISING SAID COMPOUND, PROCESS FOR PREPARING THE PHARMACEUTICAL COMPOSITION AND THERAPEUTIC USES OF THE COMPOUND - Google Patents

Abstract

This is a new compound useful as a TRPV4 antagonist, specifically the compound 1-(((5 S,7 R)-3-(5-cyclopropylpyrazin-2-yl)-7-hydroxy-2-oxo-1 -oxa-3-azaspiro[4,5]decan-7-yl)methyl)-1H-benzo[d]imidazol-6-carbonitrile, pharmaceutically acceptable salts and pharmaceutical compositions containing the compound. The compound of the invention may be useful in the treatment of a disease state selected from: atherosclerosis, disorders related to vasogenic edema, post-surgical abdominal edema, ocular edema, cerebral edema, local and systemic edema, fluid retention, sepsis, hypertension, inflammation, bone-related dysfunctions and congestive heart failure, pulmonary disorders, chronic obstructive pulmonary disorder, ventilator-induced lung injury, high-altitude induced pulmonary edema, acute respiratory distress syndrome, acute lung injury, pulmonary fibrosis, sinusitis/rhinitis, asthma , cough; including acute cough, sub-acute cough and chronic cough, pulmonary hypertension, overactive bladder, cystitis, pain, motor neuron disorders, genetic gain of function disorders, cardiovascular disease, renal dysfunction, stroke, glaucoma, retinopathy, endometriosis, premature labor, dermatitis, pruritus, pruritus in liver disease, diabetes, metabolic disorder, obesity, migraine, pancreatitis, tumor suppression, immunosuppression, osteoarthritis, Crohn's disease, colitis, diarrhea, intestinal irregularity (hyperreactivity/hyporreactivity) , fecal incontinence, irritable bowel syndrome (IBS), constipation, intestinal pain and cramps, celiac disease,(...).

Description

Field of Invention

[001] The present invention relates to a new compound useful as a TRPV4 antagonist, specifically the compound 1-(((5S,7R)-3-(5-cyclopropylpyrazin-2-yl)-7-hydroxy-2 -oxo-1-oxa-3-azaspiro[4,5]decan-7-yl)methyl)-1-benzo[d]imidazol-6-carbo-nitrile, pharmaceutically acceptable salts and pharmaceutical compositions containing the compound.

Background of the Invention

[002] TRPV4 is a member of the Transient Receptor Potential (TRP) superfamily of cation channels and is activated by heat, demonstrating spontaneous activity at physiological temperatures (Guler et al., 2002. J Neurosci 22: 64086414). Consistent with its polymodemic activation property, TRPV4 is also activated by hypotonicity and physical cellular stress/pressure (Strotmann et al., 2000. Nat Cell Biol 2: 695-702), through a mechanism involving activation of phospholipase A2, arachidonic acid and generation of epoxyeicosatrienoic acid (Vriens et al., 2004. Proc Natl Acad SciEUA 101: 396-401). Furthermore, among other proposed mechanisms, tyrosine kinase activity, as well as protein kinases A and C, may also regulate TRPV4 (Wegierski et al., 2009. J Biol Chem. 284: 292333; Fan et al. , 2009. J Biol Chem 284: 27884-91).

[003] Heart failure results in decreased ability of the left ventricle to pump blood into the peripheral circulation, as indicated by a reduced ejection fraction and/or left ventricular dilation. This increases left ventricular end-diastolic pressure, resulting in increased pulmonary blood pressures. This puts the septal barrier, which separates the circulatory aqueous environment and the alveolar air spaces of the lung, at risk. Increased pulmonary pressure results in the flow of fluid from the pulmonary circulation into the alveolar space resulting in pulmonary edema/congestion, as seen in patients with congestive heart failure.

[004] TRPV4 is expressed in the lung (Delany et al., 2001. Physiol. Genomics 4: 165-174) and its expression level is upregulated in individuals with congestive heart failure (Thorneloe et al., 2012. Sci Transl Med 4: 159ra148). TRPV4 has been shown to mediate Ca2+ entry in isolated endothelial cells and in intact lungs (Jian et al., 2009). Am J Respir Cell Mol Biol 38: 386-92). Endothelial cells are responsible for the formation of capillaries that mediate the oxygen/carbon dioxide exchange and contribute to the septal barrier in the lung. Activation of TRPV4 channels results in contraction of endothelial cells in culture and cardiovascular collapse in vivo (Willette et al., 2008. J Pharmacol Exp Ther 325:466-74), at least in part due to increased filtration at the septal barrier evoking the pulmonary edema and hemorrhage (Alvarez et al., 2006. Circ Res 99: 988-95). Indeed, filtration at the septal barrier is increased in response to increased vascular and/or airway pressures and this response is dependent on the activity of TRPV4 channels (Jian et al., 2008. Am J Respir Cell Mol Biol 38: 386 -92). Consistent with these observations, TRPV4 antagonists prevent and resolve pulmonary edema in models of heart failure (Thorneloe et al., 2012). Overall, this suggests a clinical benefit of inhibiting TRPV4 function in the treatment of pulmonary congestion associated with acute and/or chronic heart failure.

[005] Additional benefit is suggested in inhibiting TRPV4 function in lung-based pathologies, presenting symptoms including pulmonary edema/congestion, infection, inflammation, lung remodeling and/or altered airway reactivity. A genetic link between TRPV4 and chronic obstructive pulmonary disorder (COPD) was recently identified (Zhu et al., 2009. Hum. Mol GeneticS, 18: 2053-62) suggesting potential efficacy for TRPV4 modulation in the treatment of COPD with or without coincident emphysema. Enhanced TRPV4 activity is also a key factor in mechanical ventilation-induced lung injury (Hamanaka et al., 2007. Am J Physiol 293: L923-32) and it is suggested that TRPV4 activation may underlie the pathologies involved. in acute respiratory distress syndrome (ARDS), pulmonary fibrosis (Rahaman et al., 2014. J Clin Invest 124: 5225-38), cough (Bonvini et al., 2016 J Allergy Clin Immunol 138: 249-61) and asthma (Liedtke & Simon, 2004. Am J Physiol 287: 269-71). A potential clinical benefit for TRPV4 blockers in the treatment of sinusitis as well as allergic and non-allergic rhinitis is also supported (Bhargave et al., 2008). Am J Rhinol 22:7-12).

[006] TRPV4 has been shown to be involved in acute lung injury (ALI). Chemical activation of TRPV4 disrupts the alveolar septal blood barrier, potentially leading to pulmonary edema (Alvarez et al., Circ Res. 2006 Oct 27;99(9):98895). In animal models, TRPV4 antagonism attenuates lung damage induced by chemical agents and biological toxins such as HCl, chlorine gas, and platelet-activating factor (Balakrishna et al., 2014. Am J Physiol Lung Cell Mol Physiol 307: L158 -72; Morty et al., 2014. Am J Physiol Lung Cell Mol Physiol 307: L817- 21; Furthermore, TRPV4 is required in a process known to cause or worsen ALI in humans (Hamanaka et al., Am J Physiol Lung Cell Mol Physiol. 2007 Oct; 293 (4): L923-32). Overall, this suggests a clinical benefit of inhibiting TRPV4 function in the treatment of ARDS and ALI.

[007] Furthermore, TRPV4 has been implicated in recent years in several other physiological/pathophysiological processes in which TRPV4 antagonists are likely to provide significant clinical benefits. These include various aspects of pain (Todaka et al., 2004. J Biol Chem 279: 35133-35138; Grant et al., 2007. J Physiol 578: 715-733; Alessandri-Haber et al., 2006. J Neurosci 26 : 3864- 3874), to genetic motor neural disorders (Auer-Grumbach et al., 2009. Nat Genet PMID:... 20037588; Deng et al., 2009. Nat Genet PMID: 20037587; Landoure et al., 2009 . [including osteoarthritis (Muramatsu et al., 2007. J. Biol. Chem. 282: 32158-67), gain-of-function genetic mutations (Krakow et al., 2009. Am J Hum Genet 84: 307-15; Rock et al., 2008 Nat Genet 40: 999-1003) and osteoclast differentiation (Masuyama et al. 2008 Cell Metab 8: 257-65)], itch (Akiyama et al., 2016. J Invest Dermatol 136: 154- 60; Chen et al., 2016. J Biol Chem 291: 10252-62), stroke and disorders associated with cerebral edema (Li et al., 2013. Front Cell Neurosci 7: 17; Jie et al. 2015. Front Cell Neurosci e 9: 141), inflammatory bowel disorders (Vergnolle, 2014. Biochem Pharmacol 89: 157-61), various eye diseases including glaucoma and retinopathy (Monaghan et al., 2015. PloS One 10 : e0128359; Jo et al., 2016. Proc Natl Acad Sci USA 113:3885-90), and metabolic syndrome including obesity and diabetes (Ye et al., 2012 Cell 151: 96-110; Duan et al. 2015. Mol Genet Genomics 290: 1357-65).

[008] International application No. PCT/US20 12/0 42622, which has an international filing date of June 15, 2012; which also has International Publication Number WO 2013/012500 and an international publication date of January 24, 2013, describes compounds of the substituted azaspiro[4.5]decan-7-yl group that are indicated to have TRPV4 antagonistic activity and that are indicated as being useful in the treatment of various diseases including sepsis, hypertension, inflammation, bone-related disorders, heart-related disorders, lung-related disorders, pain, motor neuron disorders, renal dysfunction, osteoarthritis, Crohn's disease, colitis, irritable bowel disease (IBS), celiac disease and lactose intolerance. WO 2013/012500 does not generally or specifically disclose or claim any 7-hydroxy substituted azaspiro[4.5]decan-7-yl compound. WO 2013/012500 does not generally cover or specifically describe 1-(((5S,7R)-3-(5-cyclopropylpyrazin-2-yl)-7-hydroxy-2-oxo-1-oxa-3- azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazol-6-carbonitrile.

[009] The preference for a 7-methyl substituent in a substituted azaspiro[4.5]decan-7-yl TRPV4 antagonist was presented by: Stoy, Discovery and Optimization of Spirocarbamate TRPV4 Antagonists, Medicinal Chemistry Gordon Research Conference, August 2015 .

Summary of the Invention

[010] This invention refers to the new compound: 1-(((5S,7R)-3-(5-cyclopropyl-pyrazin-2-yl)-7-hydroxy-2-oxo-1-oxa-3- azaspiro[4,5]decan-7-yl)methyl)-1H-benzo[d]imidazol-6-carbonitrile (hereinafter referred to as “Compound A”) and pharmaceutically acceptable salts thereof. This compound is represented by the following structure: Compound A

[011] In another aspect, this invention provides Compound A for use in therapy.

[012] In another aspect, this invention provides for the use of Compound A as a TRPV4 antagonist.

[013] In another aspect, this invention provides the use of a Compound for the treatment of conditions associated with TRPV4 imbalance.

[014] In another aspect, this invention provides a method for treating a disease state selected from: atherosclerosis, disorders related to vasogenic edema, postsurgical abdominal edema, ocular edema, cerebral edema, local and systemic edema, retention of fluids, sepsis, hypertension, inflammation, bone dysfunction and congestive heart failure, pulmonary disorders, critical obstructive pulmonary disorder, ventilator-induced lung injury, high altitude-induced pulmonary edema, acute respiratory distress syndrome, acute lung injury, pulmonary fibrosis, sinusitis /rhinitis, asthma, cough; including acute cough, sub-acute cough and chronic cough, pulmonary hypertension, overactive bladder, cystitis, pain, motor neuron disorders, genetic gain of function disorders, cardiovascular disease, renal dysfunction, stroke, glaucoma, retinopathy, endometri- osis, premature labor, dermatitis, pruritus, pruritus in liver disease, diabetes, metabolic disorder, obesity, migraine, pancreatitis, tumor suppression, immunosuppression, osteoarthritis, Crohn's disease, colitis, diarrhea, intestinal irregularity (hyperreactivity/hyporreactivity), fecal incontinence, irritable bowel syndrome (IBS), constipation, intestinal pain and cramps, celiac disease, lactose intolerance, and flatulence, which comprises administering to an individual, suitably, a human individual, in need thereof of a therapeutically appropriate amount effectiveness of Compound A.

[015] In another aspect, this invention provides Compound A for use in the treatment of a pathological condition selected from: atherosclerosis, disorders related to vasogenic edema, post-surgical abdominal edema, ocular edema, cerebral edema, local edema systemic, fluid retention, sepsis, hypertension, inflammation, bone dysfunction and congestive heart failure, pulmonary disorders, chronic obstructive pulmonary disease, ventilator-induced lung injury, high-altitude induced pulmonary edema, acute respiratory distress syndrome, acute lung injury, pulmonary fibrosis, sinusitis/rhinitis, asthma, cough; including acute cough, sub-acute cough and chronic cough, pulmonary hypertension, overactive bladder, cystitis, pain, motor neuron disorders, genetic gain of function disorders, cardiovascular disease, renal dysfunction, stroke, glaucoma, retinopathy, endometriosis, premature birth, dermatitis, pruritus, pruritus in liver disease, diabetes, metabolic disorder, obesity, migraine, pancreatitis, tumor suppression, immunosuppression, osteoarthritis, Crohn's disease, colitis, diarrhea, bowel irregularity (hyperreactivity/hyporreactivity), fecal incontinence , irritable bowel syndrome (IBS), constipation, intestinal pain and cramps, celiac disease, lactose intolerance and flatulence.

[016] In another aspect, this invention provides the use of Compound A in the manufacture of a medicament for the treatment of a disease state selected from: atherosclerosis, disorders related to vasogenic edema, post-surgical abdominal edema, ocular edema, cerebral edema , local and systemic edema, fluid retention, sepsis, hypertension, inflammation, bone dysfunction and congestive heart failure, pulmonary disorders, chronic obstructive pulmonary disease, mechanical ventilation-induced lung injury, high-altitude induced pulmonary edema, distress syndrome acute respiratory, acute lung injury, pulmonary fibrosis sinusitis/rhinitis, asthma, cough; including acute cough, sub-acute cough and chronic cough, pulmonary hypertension, overactive bladder, cystitis, pain, motor neuron disorders, genetic gain of function disorders, cardiovascular disease, renal dysfunction, stroke, glaucoma, retinopathy, endometriosis, premature birth, dermatitis, pruritus, pruritus in liver disease, diabetes, metabolic disorder, obesity, migraine, pancreatitis, tumor suppression, immunosuppression, osteoarthritis, Crohn's disease, colitis, diarrhea, bowel irregularity (hyperreactivity/hyporesponsiveness), fecal incontinence , irritable bowel syndrome (IBS), constipation, intestinal pain and cramps, celiac disease, lactose intolerance and flatulence.

[017] In another aspect, the TRPV4 Antagonist can be administered alone or in conjunction with one or more other therapeutic agents, for example, agents being selected from the group consisting of endothelin receptor antagonists, angiotensin-converting enzyme inhibitors ( ACE), angiotensin II receptor antagonists, vasopeptidase inhibitors, vasopressin receptor modulators, diuretics, digoxin, beta blocker, aldosterone antagonists, iontropes, NSAIDs, nitric oxide donors, calcium channel modulators, muscarinic antagonists, anti- steroidal inflammatory agents, bronchodilators, antihistamines, leukotriene antagonists, HMG-CoA reductase inhibitors, non-selective dual antagonists of the β-adrenoreceptor and αi-adrenoceptor, type 5 phosphodiesterase inhibitors and renin inhibitors.

[018] Other aspects and advantages of the present invention are further described in the following detailed description of preferred embodiments.

Detailed Description of the Invention

[019] The present invention is directed to the new compound 1-(((5S,7R)-3-(5-cyclopropylpyrazin-2-yl)-7-hydroxy-2-oxo-1-oxa-3-azaspiro [4.5 ]decan-7-yl) methyl)- 1H - benzo [d] imidazol-6-carbonitrile (Compound A) and its pharmaceutically acceptable salts, processes for its preparation, pharmaceutical formulations comprising this compound as an active ingredient , and methods for treating a disease state associated with excess production of TRPV4 with Compound A or a pharmaceutically acceptable salt thereof, or a pharmaceutical formulation thereof.

[020] Document WO 2013/012500 does not generally or specifically disclose or claim any 7-hydroxy substituted azaspiro[4.5]decan-7-yl compound. WO 2013/012500 does not generically cover or specifically describe 1-(((5S,7R)-3-(5-cyclopropylpyrazin-2-yl)-7-hydroxy-2-oxo-1-oxa-3- azas-pyro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazol-6-carbonitrile.

[021] As used herein, "pharmaceutically acceptable" refers to compounds, materials, compositions and dosage forms that are, within the scope of medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity , irritation, or other problem or complication, compatible with a reasonable benefit/risk balance.

[022] Those skilled in the art will observe that pharmaceutically acceptable salts of Compound A can be prepared. Indeed, in certain embodiments of the invention, pharmaceutically acceptable salts of Compound A may be preferred over the respective free or salt-free compound. Therefore, the invention is further directed to pharmaceutically acceptable salts of Compound A. The invention is further directed to free or salt-free Compound.

[023] Pharmaceutically acceptable salts of the compound of the invention are readily prepared by those skilled in the art.

[024] Compound A and its pharmaceutically acceptable salts can exist in solid or liquid form. In the solid state, it can exist in crystalline or non-crystalline form, or as a mixture of these. One skilled in the art will appreciate that crystalline compounds can be formed from pharmaceutically acceptable solvates, in which solvent molecules are incorporated into the crystal lattice during crystallization. Solvates may involve non-aqueous solvents such as, but not limited to, ethanol, isopropanol, DMSO, acetic acid, ethanolamine, or ethyl acetate, or may involve water as the solvent that is incorporated into the crystal lattice. Solvates in which water is the solvent incorporated into the crystal lattice are typically referred to as "hydrates". Hydrates include stoichiometric hydrates as well as compositions containing varying amounts of water.

[025] Those skilled in the art will further observe that Compound A, and its pharmaceutically acceptable salts, can exist in crystalline form, including its various solvates, can exhibit polymorphism (that is, the ability to occur in different crystalline structures). These different crystalline forms are typically known as "polymorphs". The polymorphs have the same chemical composition, but differ in packing, geometric arrangement, and other properties descriptive of the crystalline solid state. Polymorphs, therefore, can have different physical properties, such as shape, density, hardness, deformability, stability and dissolution properties. Polymorphs typically exhibit different melting points, IR spectra, and X-ray diffraction patterns, which can be used for identification. One skilled in the art will appreciate that different polymorphs can be produced, for example, by altering or adjusting the reaction conditions or reagents used in preparing the compound. For example, changes in temperature, pressure, or solvent can result in polymorphs. Furthermore, a polymorph can spontaneously convert to another polymorph under certain conditions.

Biological activity

[026] As stated above, Compound A and pharmaceutically acceptable salts thereof are TRPV4 antagonists, and may be useful in treating disease states associated with excess TRPV4 production. Appropriately, the disease state is selected from: cerebral edema, atherosclerosis, intestinal edema-related disorders, post-surgical abdominal edema, local and systemic edema, fluid retention, sepsis, hypertension, inflammation, bone dysfunction and insufficiency congestive heart disease, pulmonary disorders, chronic obstructive pulmonary disorder, ventilator-induced lung injury, high-altitude induced pulmonary edema, acute respiratory distress syndrome, pulmonary fibrosis, sinusitis/rhinitis, asthma, overactive bladder, pain, neuron disorders motor, genetic gain of function disorders, cardiovascular disease, renal dysfunction and osteoarthritis.

[027] The biological activity of Compound A can be determined using any assay suitable for determining the activity of a candidate compound as a TRPV4 antagonist, as well as tissues and in vivo models.

[028] The biological activity of Compound A is demonstrated by the following test.

Ligand-Controlled Assay:

[029] FLIPR assay for human TRPV4 expressed in HEK293 MSRII cells

[030] Activation of the TRPV4 channel results in an influx of divalent and monovalent cations, including calcium. The resulting changes in intracellular calcium were monitored using the calcium-specific fluorescent dye Fluo-4 (MDS Analytical Technologies). HEK293 MSRII cells (human embryonic kidney 293 cells stably expressing the class II macrophage scavenger receptor) transduced with BacMam virus expressing the human TRPV4 gene at 1% final concentration were plated on a coated plate with 384-well poly-D lysine (15,000 cells/well in 50 μL culture medium containing DMEM/F12 with 15 mM HEPES, 10% FBS, 1% Penicillin-Streptomycin, and 1% L-glutamine). Cells were incubated for 24 ~ 72 hours at 37 °C and 5% CO2. The culture medium was then aspirated using a Tecan plate washer and replaced with 20 μL/well of dye loading buffer: HBSS, 500 μM Brilliant Black (MDS Analytical Technologies), and 2 μM Fluo -4 AM. The dye-loaded plates were then incubated in the dark at room temperature for 1 to 1.5 hours. 10 μL of test compounds diluted in HBSS (with 1.5 mM calcium chloride, 1.5 mM magnesium chloride, and 10 mM HEPES, pH 7.4) + 0.01% blocks were added to each individual well of the plate, incubated for 10 min at room temperature in the dark and then 10 μL of (N -((S)-1-(((R)-1-((2-cyanophenyl)sulfonyl)- agonist 3-oxoazepan-4-yl)amino)-4-methyl-1-oxopentan-2-yl)benzo[b]thiophene-2-carboxamide, Thorneloe et al., Sci. Transl. (2012), 4, 159ra148) (hereinafter: Agonist Compound) were added to have a final concentration equal to the EC80 agonist. Calcium signals were measured using FLIPRTETRA (MDS Analytical Technologies) or FLIPR384 (MDS Analytical Technologies) and inhibition of the Agonist Compound-induced calcium signal by the test compound was determined.

[031] Compound A exhibited an activity of 6.3 nM when tested in general according to the ligand-controlled assay referred to above.

Usage Methods

[032] Compound A and its pharmaceutically acceptable salts may be useful as a TRPV4 antagonist and in the treatment or prevention of a disease state selected from: atherosclerosis, disorders related to vasogenic edema, post-surgical abdominal edema, edema ocular, cerebral edema, local and systemic edema, fluid retention, sepsis, hypertension, inflammation, bone dysfunction and congestive heart failure, pulmonary disorders, chronic obstructive pulmonary disorder, ventilatory induction lung injury, high altitude induced pulmonary edema, syndrome acute respiratory distress, acute lung injury, pulmonary fibrosis, sinusitis/rhinitis, asthma, cough; including acute cough, sub-acute cough and chronic cough, pulmonary hypertension, overactive bladder, cystitis, pain, motor neuron disorders, genetic gain of function disorders, cardiovascular disease, renal dysfunction, stroke, glaucoma, retinopathy, endometriosis, premature labor, dermatitis, pruritus, pruritus in liver disease, diabetes, metabolic disorder, obesity, migraine, pancreatitis, tumor suppression, immunosuppression, osteoarthritis, Crohn's disease, colitis, diarrhea, intestinal irregularity (hyperreactivity/hyporreactivity ), fecal incontinence, irritable bowel syndrome (IBS), constipation, intestinal pain and cramps, celiac disease, lactose intolerance and flatulence. Therefore, in another aspect, the invention is directed to methods of treating such conditions.

[033] In one embodiment, this invention provides Compound A for use in the treatment of congestive heart failure.

[034] In one embodiment, this invention provides Compound A for use in treating acute lung injury.

[035] In one embodiment, this invention provides the use of Compound A in the manufacture of a medicine for the treatment of congestive heart failure.

[036] In one embodiment, this invention provides the use of Compound A in the manufacture of a medicine for the treatment of acute lung injury.

[037] Chronic cough is highly prevalent throughout the world and is highly impactful on patients' quality of life, with typical cough rates of 10 to 50 coughs per hour during waking hours. There is a hypothesis that chronic cough reflects a state of neuronal hypersensitivity involving exaggerated spinal and cortical responses to afferent sensory signals in a manner similar to chronic pain. Activation of TRPV4 channels in vivo causes the release of ATP and triggers afferent sensory signals from the lung through the binding of ATP to P2X3 channels, resulting in coughing (Bonvini, JACI, 2016). ATP levels are increased in the exhaled air of patients with cough-associated diseases, for example COPD (Basoglu, Chest, 2015). Recently, a P2X3 anatagonist demonstrated high-level efficacy in reducing chronic cough and improving quality of life scores in a phase 2 clinical trial (Abdulqawi, Lancet, 2015). These clinical data, together with data from preclinical models, suggest a role for TRPV4 receptors in the generation of cough. TRPV4 receptors are expressed on airway smooth muscle cells (McAlexander, JPET, 2014), airway epithelial cells (Delany, Physiol Genomics, 2001), and lung sensory neurons, including Ad-fibers of pathway-specific afferent neurons aerial (Bonvini, JACI, 2016). Taken together, these data suggest a potential therapeutic role for TRPV4 antagonists in cough; including acute cough, sub-acute cough and chronic cough. • Abdulqawi R, et al. Lancet. 2015 Mar 28; 385(9974):1198-1205. • Basoglu OK, et al., Chest. 2015 Aug;148(2):430-5. • Bonvini SJ, et al., J Allergy Clin Immunol. 2016 Jul;138(1):249-261.e12. • Delany NS, et al., Physiol Genomics. 2001 Jan 19;4(3): 165-74. •McAlexander MA, et al., J Pharmacol Exp Ther. 2014 Apr;349(1):118-25.

[038] Compound A is tested for its ability to treat cough in vivo in preclinical models in which cough is induced, for example, the guinea pig model cited in Bonvini et al. above. The effectiveness of Compound A is tested for its ability to treat cough; including acute cough, sub-acute cough and chronic cough in people using objective cough monitoring and specific quality of life instruments, as cited in Abdulqawi et al.

[039] The treatment methods of the present invention comprise administering a safe and effective amount of Compound A, or a pharmaceutically acceptable salt thereof, to a mammal, suitably a human being, in need thereof.

[040] As used herein, "treat", and its derivatives, in reference to a condition means: (1) improving the condition or one or more of the biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade that leads to or is responsible for the condition or (b) one or more of the biological manifestations of the condition, (3) alleviate one or more of the symptoms or effects associated with the condition, or (4) slow the progression of the condition or one or more more of the biological manifestations of the condition.

[041] The term "treat" and its derivatives refers to therapeutic therapy. Therapeutic therapy is appropriate to alleviate symptoms or to treat the first signs of the disease or its progression.

[042] Those skilled in the art will note that "prevention" is not an absolute term. In medicine, "prevention" is understood to refer to the prophylactic administration of a drug to substantially lessen the likelihood or severity of a condition or biological manifestation thereof, or to delay the onset of such a condition or biological manifestation thereof.

[043] As used herein, "safe and effective amount" in reference to Compound A, or a pharmaceutically acceptable salt thereof means an amount of the compound sufficient to treat the patient's condition, but low enough to avoid serious side effects ( in a reasonable benefit/risk ratio) within the scope of medical judgment. A safe and effective amount of the compound will vary with the particular route of administration chosen; the condition to be treated; the severity of the condition being treated; the age, size, weight and physical condition of the patient to be treated; the medical history of the patient to be treated; the duration of treatment; the nature of concomitant therapy; the desired therapeutic effect; and similar factors, but can, however, be routinely determined by one skilled in the art.

[044] As used herein, "patient" and its derivatives refer to a human being or other mammal, suitably a human being.

[045] The individual to be treated in the methods of the invention is typically a mammal in need of such treatment, preferably a human being in need of such treatment.

[046] In another aspect, the invention provides Compound A or a pharmaceutically acceptable salt thereof for use in the treatment of atherosclerosis, disorders related to vasogenic edema, post-surgical abdominal edema, ocular edema, edema cerebral, systemic and local edema, fluid retention, sepsis, hypertension, inflammation, bone-related disorders and congestive heart failure, pulmonary disorders, chronic obstructive pulmonary disease, ventilator-induced lung injury, high altitude-induced pulmonary edema, syndrome of acute respiratory distress, acute lung injury, pulmonary fibrosis, sinusitis/rhinitis, asthma, cough, including acute cough, sub-acute cough and chronic cough, pulmonary hypertension, overactive bladder, cystitis, pain, motor neuron disorders, genetic gain function disorders, cardiovascular disease, renal dysfunction, stroke, glaucoma, retinopathy, endometriosis, premature labor, dermatitis, pruritus, pruritus in liver disease, diabetes, metabolic disorder, obesity, headache, pancreatitis, tumor suppression , immunosuppression, osteoarthritis, Crohn's disease, colitis, diarrhea, intestinal irregularity (hyperreactivity/hyporesponsiveness), fecal incontinence, irritable bowel syndrome (IBS), constipation, intestinal pain and cramps, celiac disease, lactose intolerance or flatulence. Suitably, the invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof for use in the treatment of congestive heart failure. Suitably, the invention provides Compound A, or a pharmaceutically acceptable salt thereof, for use in the treatment of acute lung injuries. Suitably, the invention provides Compound A, or a pharmaceutically acceptable salt thereof, for use in the treatment of cerebral edema. Suitably, the invention provides Compound A, or a pharmaceutically acceptable salt thereof, for use in the treatment of cough; including acute cough, sub-acute cough and chronic cough.

[047] In another aspect, the invention provides the use of Compound A, or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of atherosclerosis, disorders related to vasogenic edema, post-surgical abdominal edema, ocular edema, edema cerebral, local and systemic edema, fluid retention, sepsis, hypertension, inflammation, bone dysfunction and congestive heart failure, pulmonary disorders, chronic obstructive pulmonary disease, ventilator-induced lung injury, high-altitude induced pulmonary edema, distress syndrome acute respiratory, acute lung injury, pulmonary fibrosis, sinusitis/rhinitis, asthma, cough, including acute cough, sub-acute cough and chronic cough, pulmonary hypertension, overactive bladder, cystitis, pain, motor neuron disorders, genetic gain of function, cardiovascular diseases, renal dysfunction, stroke, glaucoma, retinopathy, endometriosis, premature labor, dermatitis, pruritus, pruritus in liver disease, diabetes, metabolic disorder, obesity, migraine, pancreatitis, tumor suppression, immunosuppression, osteoarthritis , Crohn's disease, colitis, diarrhea, intestinal irregularity (hyperreactivity/hyporreactivity), fecal incontinence, irritable bowel syndrome (IBS), constipation, intestinal pain and cramping, celiac disease, lactose intolerance or flatulence. Suitably, the invention provides the use of a compound of Formula (I) or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of congestive heart failure. Suitably, the invention provides the use of Compound A, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of acute lung injuries. Suitably, the invention provides the use of Compound A, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of cerebral edema.

[048] Compound A, or a pharmaceutically acceptable salt thereof, can be administered by any suitable route of administration, including both systemic and topical administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, rectal administration and inhalation administration. Parenteral administration refers to routes of administration other than enteral, transdermal, or inhalation, and is typically by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion. Inhalation refers to administration into the patient's lungs, whether inhaled through the mouth or through the nasal passages. Topical administration includes application to the skin, as well as intraocular, otic, intravaginal, and intranasal administration. Properly, administration is oral. Properly, administration is intravenous. Properly, administration is by inhalation.

[049] Compound A, or a pharmaceutically acceptable salt thereof, can be administered once or according to a dosage regimen in which a number of doses are administered at varying time intervals over a given period of time. For example, doses may be administered one, two, three or four times per day. Doses may be administered until the desired therapeutic effect is achieved or indefinitely to maintain the desired therapeutic effect. Suitable dosage regimens for the compound depend on the properties of the compound, such as absorption, distribution, and half-life, which can be determined by those skilled in the art. Furthermore, suitable dosage regimens, including the duration of such regimens are administered, for the compound of the invention depend on the condition being treated, the severity of the condition being treated, the age and physical condition of the patient being treated, the medical history of the patient to be treated, the nature of the concomitant therapy, the desired therapeutic effect and similar factors within the knowledge and skill of one skilled in the art. It would further be understood by those skilled in the art that suitable dosage regimens may require adjustments given an individual patient's response to the dosage regimen or over time as the individual patient needs to change.

[050] Typical daily dosages may vary depending on the particular route of administration chosen. Typical dosages for oral administration range from 1 mg to 1000 mg per person per dose. Preferred dosages are 1 to 500 mg once daily or twice daily per person.

[051] Additionally, Compound A, or a pharmaceutically acceptable salt thereof, can be administered as a prodrug. As used herein, a "prodrug" of a compound of the invention is a functional derivative of the compound that, upon administration to a patient, eventually releases the compound of the invention in vivo. Administration of a compound of the invention as a prodrug may allow the skilled artisan to do one or more of the following: (a) modify the initiation of the compound in vivo; (b) modify the duration of action of the compound in vivo; (c) modify the transport or distribution of the compound in vivo; (d) modify the solubility of the compound in vivo; and (e) overcome or overcome a side effect or other difficulty encountered with the compound. Typical functional derivatives used to prepare prodrugs include modifications of the compound that are chemically or enzymatically cleaved in vivo. Such modifications, which include the preparation of phosphates, amides, ethers, esters, thioesters, carbonates and carbamates, are well known to those skilled in the art.

Compositions

[052] Compound A, or a pharmaceutically acceptable salt thereof, will normally, but not necessarily, formulate into a pharmaceutical composition prior to administration to a patient. Thus, in another aspect, the invention is directed to pharmaceutical compositions comprising Compound A and one or more pharmaceutically acceptable excipients.

[053] The pharmaceutical compositions of the invention can be prepared and packaged in bulk form, in which a safe and effective amount of Compound A can be extracted and then administered to the patient, such as with powders or syrups. Alternatively, the pharmaceutical compositions of the invention may be prepared and packaged in unit dosage form, wherein each physically discrete unit contains a safe and effective amount of Compound A. When prepared in unit dosage form, the pharmaceutical compositions of the invention typically contain from 1 mg to 1000 mg of Compound A. Furthermore, the pharmaceutical compositions of the invention may optionally further comprise one or more additional pharmaceutically active compounds.

[054] As used herein, "pharmaceutically acceptable excipient" means a pharmaceutically acceptable material, composition or vehicle involved in giving shape or consistency to the pharmaceutical composition. Each excipient must be compatible with the other ingredients of the pharmaceutical composition when mixed, so that interactions that substantially reduce the effectiveness of the compound of the invention when administered to a patient and interactions that would result in pharmaceutical compositions that are not pharmaceutically acceptable are avoided. Furthermore, each excipient must, of course, be of sufficiently high purity to make it pharmaceutically acceptable.

[055] Compound A and the pharmaceutically acceptable excipient or excipients will typically be formulated in a dosage form adapted for administration to the patient by the desired route of administration. For example, dosage forms include those adapted for (1) oral administration such as tablets, capsules, caplets, pills, lozenges, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets and sachets; (2) parenteral administration, such as sterile solutions, suspensions, and powders for reconstitution; (3) transdermal administration, such as transdermal patches; (4) rectal administration such as suppositories; (5) inhalation, such as dry powders, aerosols, suspensions and solutions; and (6) topical administration, such as creams, ointments, lotions, solutions, pastes, sprays, foams and gels.

[056] Suitable pharmaceutically acceptable excipients will vary depending on the particular dosage form chosen. Additionally, suitable pharmaceutically acceptable excipients can be chosen for the particular function they can serve in the composition. For example, certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the production of uniform dosage forms. Certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate the production of stable dosage forms. Certain pharmaceutically acceptable excipients may be chosen for their ability to facilitate transport or transport of Compound A once administered to the patient from one organ, or portion of the body, to another organ or portion of the body. Certain pharmaceutically acceptable excipients may be chosen for their ability to improve patient compliance.

[057] Suitable pharmaceutically acceptable excipients include the following types of excipients: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweeteners, flavorings, flavorings, masking agents, coloring agents, anti-caking agents, hemectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants and buffering agents. One skilled in the art will appreciate that certain pharmaceutically acceptable excipients may serve more than one function and may serve alternative functions depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation.

[058] Qualified experts have the knowledge and expertise in the art that allows them to select suitable pharmaceutically acceptable excipients in appropriate amounts for use in the invention. Furthermore, there are several resources that are available to one skilled in the art that describe pharmaceutically acceptable excipients and may be useful in selecting suitable pharmaceutically acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).

[059] The pharmaceutical compositions of the invention are prepared using techniques and methods known to those skilled in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).

[060] In one aspect, the invention is directed to a solid oral dosage form, such as a tablet or capsule, comprising a safe and effective amount of compound A and a diluent or filler. Suitable diluents and fillers include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g., corn starch, potato starch and pregelatinized starch), cellulose and its derivatives (e.g., microcrystalline cellulose), calcium sulfate and dibasic calcium phosphate. The solid oral dosage form may further comprise a binder. Suitable binders include starch (e.g. corn starch, potato starch and pregelatinized starch), gelatin, acacia, sodium alginate, alginic acid, tragacanth, guar gum, povidone and cellulose and its derivatives (e.g. cellulose microcrystalline). The solid oral dosage form may further comprise a disintegrant. Suitable disintegrants include crospovidone, sodium starch glycolate, croscarmelose, alginic acid and sodium carboxymethyl cellulose. The solid oral dosage form may further comprise a lubricant. Suitable lubricants include stearic acid, magnesuine stearate, calcium stearate and talc.

[061] Included in the present invention is a process for preparing a pharmaceutical composition containing a pharmaceutically acceptable carrier and Compound A which process comprises placing Compound A in association with a pharmaceutically acceptable carrier.

[062] Compound A, or a pharmaceutically acceptable salt thereof, can be administered alone or together with one or more other therapeutic agents, said agents being selected from the group consisting of endothelin receptor antagonists, endothelin-converting enzyme. angiotensin (ACE), angiotensin II receptor antagonists, vasopeptidase inhibitors, vasopressin receptor modulators, diuretics, digoxin, beta blocker, aldosterone antagonists, iontropes, NSAIDs, nitric acid donors, calcium channel modulators, muscarinic antagonists, steroidal anti-inflammatory drugs, bronchodilators, antihistamines, leukotriene antagonists, HMG-CoA reductase inhibitors, non-selective dual β-adrenoceptor and α-adrenoceptor antagonists, phosphodiesterase type 5 inhibitors and renin inhibitors .

[063] The invention also provides a pharmaceutical composition comprising from 0.5 to 1,000 mg of Compound A, or a pharmaceutically acceptable salt and from 0.5 to 1,000 mg of a pharmaceutically acceptable excipient.

Examples

[064] The following examples illustrate the invention. These examples are not intended to limit the scope of the present invention, but rather to provide guidance to one skilled in the art for preparing and using the compound, compositions and methods of the present invention. Although particular embodiments of the present invention are described, one skilled in the art will appreciate that various changes and modifications can be made without departing from the spirit and scope of the invention.

[065] The following abbreviations and terms have the meanings indicated at all times:

Example 1

[066] Preparation of: 1-(((5S,7R)-3-(5-cyclopropylpyrazin-2-yl)-7-hydroxy-2-oxo- 1-oxa-3-azaspiro[4.5]decan-7- yl)methyl)-1H-benzo[d]imidazol-6-carbonitrile (Compound A) Scheme 1 Scheme 2 Scheme 3

Stage 1: 1,4-dioxaspiro[4.5] decan-7-one

[067] A solution of cyclohexane-1,3-dione (500 g, 4,459 mmol), 4Å molecular sieves (500 g, 4,459 mmol) and p-toluenesulfonic acid (254 g, 1,338 mmol) in dry ethylene glycol ( 2L) was stirred under nitrogen at room temperature for 4 hours. The reaction mixture was diluted with saturated NaHCO3 solution (1L) to adjust the basic pH, and the basic mixture was extracted with ethyl acetate (3 x 1 L). The combined organic extracts were washed with brine solution (500 ml) and dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the title compound (280 g, 1732 mmol, 38.8% yield) as a yellow liquid. . LCMS (m/z) 157.1 (M+H+).

Stage 2: 1,6,9-trioxadispiro[2.1.4.3]dodecane

[068] A solution of 1,4-dioxaspiro[4,5]decan-7-one (280g, 1,793 mmol) in dimethylsulfoxide (1.5L) under nitrogen at 20 °C was treated with trimethylsulfoxonium iodide (395 g, 1,793 mmol) and potassium tert-butoxy (221 g, 1,972 mmol) and the reaction mixture was stirred for 16 hours before being diluted with water and extracted with ethyl acetate (3 x 2 L). The combined organic extracts were washed with brine (2 x 500 ml), dried over Na2SO4, filtered, concentrated under reduced pressure and purified by silica gel column chromatography (15% ethyl acetate/petroleum ether) to yield the compound of the title (170 g, 962 mmol, 53.6% yield) as a colorless oil. GCMS (m/z) 141.2, 170.2.

Stage 3: 7-(aminomethyl)-1,4-dioxaspiro[4.5]decan-7-ol

[069] A mixture of 1,6,9-trioxadispiro[2.1.4.3]dodecane (125 g, 734 mmol) and 7N ammonia in MeOH (2.96 L, 20.7 mol) was stirred at room temperature for 72 hours. The reaction mixture was concentrated under reduced pressure to obtain a crude solid, which was triturated with pentane and dried under vacuum to obtain the title compound (100 g, 486 mmol, 66.2% yield) as an off-white solid. GC MS (m/z) 187.2.

Stage 4: 3-(((7-hydroxy-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)-4-nitro-benzonitrile

[070] A mixture of 7-(aminomethyl)-1,4-dioxaspiro[4,5]decan-7-ol (100 g, 534 mmol), 3-fluoro-4-nitrobenzonitrile (89 g, 534 mmol) and Potassium carbonate (148 g, 1068 mmol) in acetonitrile (1L) was stirred at room temperature under nitrogen for 18 hours. The reaction mixture was diluted with saturated NH4Cl solution (1.5L) and extracted with ethyl acetate (2 x 2L). The combined organic extracts were washed with brine (2 x 200 ml), dried over Na2SO4 and concentrated under reduced pressure. The crude material was triturated with 2% MeOH/CH2Cl2 (200 ml), filtered and washed with diethyl ether (300 ml) to obtain the title compound (120 g, 356 mmol, 66.7% yield) as a orange solid. LC MS (m/z) 334.0 (M+H+).

Stage 5: 4-amino-3-(((7-hydroxy-1,4-dioxaspiro[4.5]decan-7-yl)methyl)amino)benzonitrile

[071] A solution of 3-(((7-hydroxy-1,4-dioxaspiro[4.5]decan-7-yl) methyl)amino)-4-nitrobenzonitrile (120 g, 360 mmol) and ammonium chloride (193 g, 3600 mmol)) in ethanol (1500 ml) and water (500 ml) under nitrogen was treated with iron (201 g, 3600 mmol). The reaction mixture was stirred at room temperature for 2 hours and then heated at 85°C for 6 hours before being cooled to room temperature and concentrated under reduced pressure to remove ethanol. The remaining aqueous mixture was extracted with CH2Cl2 (2 x 1500 ml). The combined organic extracts were washed with NaHCO3 (1000 ml), brine (150 ml), dried over Na2SO4, filtered and concentrated under reduced pressure to obtain the title compound (110 g, 312 mmol, 87% yield) as a foam. pale yellow. LC MS (m/z) 304.0 (M+H+).

Stage 6: 1-((7-hydroxy-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazol-6-carbonitrile

[072] A solution of 4-amino-3 -(((7-hydroxy-1,4-dioxaspiro[4.5]decan-7-l)methyl)amino)benzonitrile (110 g, 312 mmol) and orthoformate Trimethyl (34.5 mL, 312 mmol) was treated with formic acid (11.96 mL, 312 mmol) and stirred at 80°C for 4 hours before being concentrated under reduced pressure. The residue was diluted with ethyl acetate (1.5 L) and washed with NaHCO3 solution (1 L), water (100 ml), and brine (100 ml). The organic layer was concentrated under reduced pressure. The material was triturated with acetone (100 mL), diethyl ether (3 x 300 mL) and hexane and dried to provide the title compound (80 g, 254 mmol, 81% yield) as an off-white solid. LCMS (m/z) 314.0 (M+H+).

Stage 7: 1-((1-hydroxy-3-oxocyclohexyl)-methyl)-1 H -benzo [d]imidazol-6-carbonitrile

[073] A solution of 1-((7-hydroxy-1,4-dioxaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazol-6-carbonitrile (60 g, 191 mmol) in Acetone (600 mL) and water (200 mL) was treated with 1N aqueous HCl (94 mL, 94 mmol) and stirred at room temperature for 24 hours. The reaction mixture was diluted with ethyl acetate and water and the layers were separated. The organic layer was washed with water (500 ml). The combined aqueous extracts were washed with ethyl acetate (3 x 800 ml). The combined organic extracts were washed with brine, dried over Na2SO4, and concentrated under reduced pressure. The resulting material was triturated with diethyl ether (550 ml), dried and then suspended in CH2Cl2 (500 ml) and stirred for 16 hours. The solid was collected by filtration and purified by silica gel column chromatography (0 to 4% MeOH/CH2Cl2) to give the title compound (30 g, 109 mmol, 57% yield) as an off-white solid. LC MS (m/z) 270.2 (M+H+).

Stage 8: (+/-) 1-(((3S, 5R)-5-hydroxy-1-oxaspiro[2.5]octan-5-yl) methyl)-1H-benzo[d]imidazol-6-carbonitrile (mixture racemic of cis enantiomers)

[074] A solution of 1-((1-hydroxy-3-oxocyclohexyl)methyl)-1H-benzo[d]imidazol-6-carbonitrile (30 g, 111 mmol) and trimethylsulfoxonium iodide (27, 0 g, 123 mmol) in dimethyl sulfoxide (200 ml) under nitrogen was treated with potassium tert-butide (13.75 g, 123 mmol) and stirred at room temperature for 1 hour. The reaction mixture was poured into ice-water (500 mL) and extracted with ethyl acetate (6 X 550 mL). The combined organic extracts were washed with water (2 x 100 ml) and brine (100 ml), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was triturated with 2:8 MeCN: diethyl ether (2 x 100 ml) and the solid was collected by filtration, washed with diethyl ether, and purified by silica gel column chromatography (0 to 3% MeOH/ CH2Cl2) to yield the title compound (12.5 g, 43.2 mmol, 38.8% yield) as a white solid. LC MS (m/z) 284.3 (M+H+).

Stage 9: 1-(((3S, 5R)-5-hydroxy-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazol-6-carbonitrile

[075](+/-) 1-(((3S,5R)-5-hydroxy-1-oxaspiro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazol-6-carbonitrile (mixture racemic cis enantiomers) (45 g, 159 mmol) was dissolved in 1:1 CH2Cl2:EtOH (900 ml) and the enantiomers were resolved by chiral supercritical fluid chromatography (SFC) using a ChiralPak IC column (30 x 250 mm, 5 μm) under isocratic conditions (35% IPA co-solvent, flow rate 60 grams/minute) to obtain the title compound (14.08 g, 47.22 mmol, 29.7% , > ee 99%) as a pale orange solid. LCMS (m/z) 284.1 (M+H+).

Stage 10: 1-(((5S,7R)-7-hydroxy-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazole- 6-carbonitrile

[076] A mixture of ethyl carbamate (15.47 g, 174 mmol), lithium tert-butide (1M in hexanes) (17.37 mL, 17.37 mmol) and N-methyl-2-pyrrolidone (174 mL) was stirred at room temperature for 5 minutes before 1-(((3S,5R)-5-hydroxy-1-oxas-piro[2.5]octan-5-yl)methyl)-1H-benzo[d]imidazole -6-carbonitrile (9.84 g, 34.7 mmol) was added. The mixture was stirred at 75°C for 15 hours, then cooled to room temperature and poured into 400 ml of water (heating of the mixture was observed). The resulting light orange suspension was stirred overnight and the solid was collected by filtration to give the title compound (6.745 g, 17.57 mmol, 50.6% yield) as a pale orange solid. LCMS (m/z) 327.0 (M+H+).

Stage 11: 2-bromo-5-cyclopropylpyrazine

[077] 2-Bromo-5-cyclopropylpyrazine was ordered from CombiPhos catalysts (CS2504) or synthesized using the following preparation. A solution of 2,5-dibromopyrazine (60 g, 252 mmol) in toluene (600 ml) was stirred and degassed with nitrogen for 10 minutes before a solution of K2CO3 (87 g, 631 mmol) in water (80 ml) was added. ) was added. The mixture was degassed for an additional 5 minutes, then cyclopropylboric acid (28.2 g, 328 mmol), palladium(II) acetate (2.83 g, 12.61 mmol), and PdCl2 (dppf) CH2Cl2 were added. adduct (10.30 g, 12.61 mmol) were added and the reaction mixture was stirred at 80°C overnight. The reaction mixture was filtered through celite and the filtrate was diluted with ethyl acetate (400 mL) and washed with water (2 x 200 mL) and brine. The organic layer was dried over Na2SO4, filtered, concentrated under reduced pressure and purified by silica gel chromatography (3% ethyl acetate/hexanes) to give a yellow oil, which was dissolved in 1:9 CH2Cl2:pentane (50 ml) and cooled to -20 °C. A precipitate formed and the resulting suspension was stirred for 10 minutes before the solvents were decanted and the solid dried. The CH2Cl2:pentane process was repeated, the solid obtained was dried under vacuum to give the title compound (18.35 g, 88 mmol, 34.9% yield) as a white crystalline solid. GCMS (m/z) 197.0/199.0.

Stage 12: 1-(((5S,7R)-3-(5-cyclopropylpyrazine-2-yl)-7-hydroxy-2-oxo-1-oxa-3- azaspiro[4.5]decan-7-yl)methyl )-1H-benzo[d]imidazol-6-carbonitrile

[078] A mixture of 1-(((5S,7R)-7-hydroxy-2-oxo-1-oxa-3-azaspiro[4.5]decan-7-yl)methyl)-1H - benzo[d]imidazole -6-carbonitrile (6.736 g, 20.64 mmol), 2-bromo-5-cyclopropylpyrazine (6.16 g, 31.0 mmol), and potassium phosphate, tribasic (8.76 g, 41.3 mmol) in 1,4-dioxane (138 mL) treated with N1,N2-dimethylethane-1,2-diamine (1.819 g, 20.64 mmol) and copper (I) (1.965 g, 10.32 mmol) and stirred at 95°C for 16 hours. The mixture was then cooled to room temperature, diluted with CH2Cl2 (200 ml), water (150 ml) and 7N NH3 in MeOH (10 ml; to remove copper impurities). The layers were separated, and the aqueous layer was extracted with CH2Cl2 (2 X 50 ml). The combined CH2Cl2 was washed with an 8:2 mixture of water and 7N NH3 in MeOH (3 X 50 ml), dried over Na2SO4, filtered, concentrated under reduced pressure, and purified by silica gel chromatography (0 to 10 % methanol/dichloromethane) to give a pale orange solid. This solid was stirred in 60 ml of EtOH at 50°C for 30 minutes and then at room temperature overnight before being collected by filtration to give a white solid. This solid was stirred in 40 ml of 50°C MeCN for 30 minutes, then at room temperature for 1 hour and then collected by filtration to provide the title compound (3.395 g, 7.26 mmol, 35.2% yield) as a solid white crystalline. LC MS (m/z) 445.3 (M+H+). 1H NMR (400 MHz, DMSO- d6) δ ppm: 9.12 (d, J = 1.5 Hz, 1 H), 8.39 (s, 1 H), 8.37 (d, J = 1, 5 Hz, 1 H), 8.32 (d, J = 1.2 Hz, 1H), 7.81 (d, J = 8.4 Hz, 1H), 7.56 (dd, J = 8.4 , 1.2 Hz, 1H), 4.66 (s, 1H), 4.26 (d, J = 14.6 Hz, 1H), 4.22 (d, J = 14.6 Hz, 1H), 3.92 (d, J = 10.2 Hz, 1H), 3.82 (d, J = 10.2 Hz, 1H), 2.12-2.21 (m, 1H), 1.78-2 .04 (m, 4H), 1.35-1.64 (m, 4H) 0.96 - 1.02 (m, 2H), 0.86 - 0.91 (m, 2H).

Example 2 - Capsule Composition

[079] An oral dosage form for administering the present invention is produced by depositing a standard two-piece hard gelatin capsule with the ingredients in the proportions shown in Table 1, below. Table 1

Example 3 - Injectable Parenteral Composition

[080] An injectable form for administering the present invention is produced by stirring 1.7% by weight of 1-(((5S,7R)-3-(5-cyclopropylpyrazin-2-yl)-7-hydroxy-2 - oxo-1-oxa-3-azaspiro[4,5]decan-7-yl) methyl)-1 H -benzo[d]imidazol-6-carbonitrile (Compound A) in 10% by volume of propylene glycol in water.

Example 4 - Table Composition

[081] Sucrose, calcium sulfate dihydrate and Compound A as shown in Table 2 below, are mixed and granulated in the proportions shown with a 10% gelatin solution. The wet granules are filtered, dried, mixed with starch, talc and stearic acid, sieved and compressed into a tablet. Table 2

[082] Although the preferred embodiments of the invention are illustrated by the above, it should be understood that the invention is not limited to the precise instructions disclosed herein and that the right to all modifications falling within the scope of the following claims is reserved.

Claims (16)

1. Compound CHARACTERIZED by the fact that it is: 1-(((5S,7R)-3-(5-cyclopropylpyrazin-2-yl)-7-hydroxy-2-oxo-1-oxa-3-azaz-pyro[ 4.5]decan-7-yl)methyl)-1H-benzo[d]imidazol-6-carbonitrile: or a pharmaceutically acceptable salt thereof. 2. Compound, according to claim 1, CHARACTERIZED by the fact that it is 1-(((5S,7R)-3-(5-cyclopropylpyrazin-2-yl)-7-hydroxy-2-oxo-1-oxa -3-azas- pyro[4.5]decan-7-yl)methyl)-1H-benzo[d]imidazol-6-carbonitrile: 3. Compound according to claim 1, CHARACTERIZED by the fact that it is a pharmaceutically acceptable salt of 1-(((5S,7R)-3-(5-cyclopropylpyrazin-2-yl)-7-hydroxy-2- oxo-1-oxa-3-azaspiro[4,5]decan-7-yl)methyl)-1H-benzo[d]imidazol-6-carbonitrile: 4. Pharmaceutical composition CHARACTERIZED by the fact that it comprises a compound, as defined in any one of claims 1 to 3, and a pharmaceutically acceptable carrier. 5. Pharmaceutical composition, according to claim 4, CHARACTERIZED by the fact that the composition is in tablet form. 6. Pharmaceutical composition, according to claim 4, CHARACTERIZED by the fact that the composition is in an intravenous form. 7. Process for preparing a pharmaceutical composition containing a pharmaceutically acceptable carrier and a compound, as defined in any one of claims 1 to 3, CHARACTERIZED by the fact that the process comprises placing the compound in association with a pharmaceutically acceptable carrier. 8. Use of a compound or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 3, CHARACTERIZED by the fact that it is for the manufacture of a medicament for the treatment or alleviation of the severity of a disease state selected from: atherosclerosis, disorders related to vasogenic edema, post-surgical abdominal edema, ocular edema, cerebral edema, local and systemic edema, fluid retention, sepsis, hypertension, inflammation, bone-related dysfunctions and congestive heart failure, pulmonary disorders, chronic obstructive pulmonary disorder, ventilator-induced lung injury, high-altitude induced pulmonary edema, acute respiratory distress syndrome, acute lung injury, pulmonary fibrosis, sinusitis/rhinitis, asthma, cough, acute cough, sub-acute cough, chronic cough, pulmonary hypertension, overactive bladder, cystitis, pain, motor neuron disorders, genetic gain of function disorders, cardiovascular disease, renal dysfunction, stroke, glaucoma, retinopathy, endometriosis, premature labor, dermatitis, pruritus, pruritus disease liver, diabetes, metabolic disorder, obesity, migraine, pancreatitis, tumor suppression, immunosuppression, osteoarthritis, Crohn's disease, colitis, diarrhea, intestinal irregularity (hyperreactivity/hyporreactivity), fecal incontinence, irritable bowel syndrome (IBS), constipation, intestinal pain and cramps, celiac disease, lactose intolerance and flatulence. 9. Use, according to claim 8, CHARACTERIZED by the fact that the compound is formulated to be administered orally. 10. Use, according to claim 8, CHARACTERIZED by the fact that the compound is formulated to be administered intravenously. 11. Use, according to claim 8, CHARACTERIZED by the fact that the compound is formulated to be administered by inhalation. 12. Use, according to claim 8, CHARACTERIZED by the fact that the disease is congestive heart failure. 13. Use, according to claim 8, CHARACTERIZED by the fact that the disease is acute lung injury. 14. Compound or a pharmaceutically acceptable salt thereof, according to any one of claims 1 to 3, CHARACTERIZED by the fact that it is for use in therapy. 15. Use of a compound or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 3, CHARACTERIZED by the fact that it is for the manufacture of a medicament for the treatment or alleviation of the severity of congestive heart failure. 16. Use of a compound or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 3, CHARACTERIZED by the fact that it is for the manufacture of a medicament for the treatment or alleviation of the severity of acute lung injury.