Fluorinated adamantyl p2x7 receptor antagonists and uses thereof

Abstract

The invention relates generally to a series of adamantyl and mono-, di-, and tri- substituted adamantyl P2X7 receptor antagonists of Formula (I) and their use in the treatment of inflammatory conditions such as cardiovascular disease, wherein each of R1, R2 and R3 may be H or F, with the proviso that R1, R2 and R3 cannot each be H, and pharmaceutically-acceptable salts thereof:

Description

FLUORINATED ADAMANTYL P2X7 RECEPTOR ANTAGONISTS AND USES THEREOF Related Application

[0001] The present application claims convention priority to Australian provisional patent application 2023902401, filed on 28 July 2023. The content of AU’401 is incorporated herein by reference in its entirety. Field of the Invention

[0002] The present invention relates to a series of adamantyl and mono-, di-, and tri- substituted adamantyl P2X7 receptor antagonists of Formula (I), wherein each of R1,R2and R3 may be H or F, with the proviso that R1, R2 and R3 cannot each be H, and pharmaceutically-acceptable salts thereof: (I).

[0003] Other forms of the invention relate to the synthesis and medical uses of such compounds, in particular, for the treatment of cardiovascular diseases.

[0004] Although the present invention will be described hereinafter with reference to its preferred embodiment, it will be appreciated by those of skill in the art that the spirit and scope of the invention may be embodied in many other forms. Background of the Invention

[0005] Any discussion of the prior art throughout the specification should in no way be considered as an admission that such prior art is widely known or forms part of common general knowledge in the field.

[0006] The P2X7 receptor (P2X7R) is a purinergic receptor which has been implicated in numerous central nervous system (CNS) disorders due to its central role in the inflammatory response. Activation of the P2X7R releases pro-inflammatory cytokines such as interleukin 1β, which have been shown to underlie the pathogenesis of many inflammatory disorders. P2X7 signaling has been heavily implicated in atherosclerosis and atherothrombosis throughin vitro and in vivo experiments, with blockade of receptor-mediated signaling mitigating atherosclerosis, hypertension and diabetic retinopathy, as well as dilated cardiomyopathy and arrhythmia post myocardial infarction.

[0007] Recent studies have shown that the P2X7R also plays a role in cardiovascular diseases (CVDs), with a special focus on ischemic injury.

[0008] International patent publication WO 2001 / 044170, entitled “Adamantane derivatives”, to AstraZeneca AB, discloses molecule “AZD9056”, as shown below:

[0009] WO’170 teaches adamantane derivatives including AZD9056, a process for their preparation, pharmaceutical compositions containing them, a process for preparing the pharmaceutical compositions, and their use in effecting immunosuppression (e.g., in the treatment of rheumatoid arthritis, irritable bowel disease, atherosclerosis, psoriasis, pulmonary disease, e.g., COPD or bronchitis, or diseases of the central nervous system, e.g., Alzheimer’s disease or stroke).

[0010] Subsequently, AZD9056 has been documented in clinical trials for rheumatoid arthritis (Keystone, E. C.; Wang, M. M.; Layton, M.; Hollis, S.; McInnes, I. B., Ann. Rheum. Dis.2012, 71 (10), 1630–1635.10.1136 / Annrheumdis-2011-143578). While no efficacy for rheumatoid arthritis was established, phase IIb found orally-administered AZD9056 was well tolerated up to 400 mg / day. Further, the present inventors understand that is one trial currently underway for Major Depressive Disorder (Inflammation) by Janssen (Phase 2 clinical trial with API JNJ-54175446).

[0011] International patent publication WO 2006 / 083214, entitled “Pharmaceutical composition comprising a P2X7 receptor antagonist and a HMG-CoA reductase inhibitor”, to AstraZeneca AB, teaches a pharmaceutical composition, product or kit comprising a first active ingredient which is a P2X7 receptor antagonist and a second active ingredient which is a HMG-CoA reductase inhibitor. The P2X7 receptor antagonist is optionally an adamantly derivate, such as AZD9056, and the HMG-CoA reductase inhibitor a statin. The invention is intended for use in the treatment of cardiovascular disorders, especially atherosclerosis.

[0012] Chinese patent application CN 103520721, entitled “Application of P2X7 antagonistoxATP in preparation of medicament for treating myocardial ischemia injuries and hypertension / sympathetic nervous system diseases”, to Nanchang University, discloses a P2X7 antagonist in preparation of medicament for treating myocardial ischemia injuries and hypertension / sympathetic nervous system diseases. The document discloses a P2X7 receptor antagonist oxATP which can be used for reducing sympathetic nervous reflex caused by myocardial ischemia injuries and avoiding the phenomenon that myocardial ischemia and hypoxia are further aggravated due to cardiac sympathetic erethism, so that the myocardial ischemia injuries are relieved or alleviated. The hypertension can be prevented and cured by inhibiting cervical sympathetic ganglion erethism transmission. Sympathetic nervous diseases can be prevented and cured by inhibiting sympathetic erethism and related pathological processes. The mechanism of action of the oxATP in preventing and treating myocardial ischemia injuries, hypertension and sympathetic nervous lesions is as follows: the sympathetic excitation transmission mediated by cervical sympathetic ganglion P2X7 accepter is blocked, and an effect of preventing and treating the myocardial ischemia injuries, hypertension and sympathetic nervous diseases is generated.

[0013] Chinese patent application CN 112245585, entitled “Application of P2X7 receptor inhibitor in preparation of Ang II (angiotension II) induced cardiomyopathy prevention and treatment drug”, to The First Affiliated Hospital of Wenzhou Medical University, teaches the application of a P2X7 receptor inhibitor in preparing a drug for preventing and treating Ang II (angiotensin II) induced cardiomyopathy, which is based on the physiological action of a P2X7 receptor and the basis of other field researches, clarifies the influence and corresponding mechanism of the P2X7 receptor on the occurrence and development of Ang II-induced cardiomyopathy, discloses a new target point for regulating and controlling Ang II-induced myocardial remodeling, and provides a new revelation for preventing and treating hypertensive heart remodeling.

[0014] International patent publication WO 2020 / 037350, entitled “Adamantanyl- substituted benzamide compounds and their use as P2X7 receptor antagonists”, to the University of Sydney, teaches adamantanyl-substituted benzamide compounds and their use as antagonists of the P2X7 purinoreceptor. The invention further relates to methods for the treatment of disease and conditions associated with the P2X7purinoreceptor. The disease or condition may be an inflammatory disease or condition such as a neuroinflammatory disease or a neurodegenerative disease, or it may be a disease of the central nervous system. In one embodiment, the disease or condition is coronary artery disease, cardiovascular disease or atherosclerosis, more particularly acute coronary syndrome, coronary artery disease,myocarditis, pericarditis, myocardial ischemia and reperfusion injury.

[0015] International patent publication WO 2023 / 092175, entitled “Novel P2X7 receptor antagonists”, to the University of Sydney, teaches a series of novel heterocyclic adamantyl and F3-adamantyl cyanoguanidine P2X7 receptor. The compounds or pharmaceutically acceptable salts thereof have potential medical use in the treatment or prevention of conditions such as stroke; amyotrophic lateral sclerosis; multiple sclerosis; Alzheimer s disease; Huntington s disease, atherosclerosis, diabetic retinopathy; dilated cardiomyopathy; ischemic injury and left ventricular hypertrophy post myocardial infarction.

[0016] It is an object of the present invention to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative.

[0017] The development of a P2X7R inhibitor capable of reducing central and peripheral proinflammatory signaling has been a major goal of numerous pharmaceutical and academic groups. CNS-penetrant P2X7R antagonists have yet to proceed past clinical trials. Moreover, in cardiovascular diseases (CVD), there is no available drug for P2X7R modulation.

[0018] The present invention relates to the use of known P2X7R antagonist AZD9056 to combat atherosclerosis and CVD. The present inventors have explored the chemical space around the known adamantanyl benzamide series and have generated a series of novel P2X7R antagonists, named the “PKT400” series.

[0019] Use of AZD9056 in the treatment of cardiovascular diseases and synthesis of the novel PKT400 series for use in treating inflammatory conditions such as CVD represent potentially significant advances in the development of a drug-like P2X7R antagonist for treatment of atherosclerosis and CVD.

[0020] Although the invention will be described with reference to specific examples it will be appreciated by those skilled in the art that the invention may be embodied in many other forms. Definitions

[0021] In describing and defining the present invention, the following terminology will be used in accordance with the definitions set out below. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments of the invention only and is not intended to be limiting.

[0022] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one having ordinary skill in the art to which theinvention pertains.

[0023] Unless the context clearly requires otherwise, throughout the description and the claims, the words “comprise”, “comprising” and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”.

[0024] As used herein, the phrase “consisting of” excludes any element, step, or ingredient not specified in the claim. When the phrase “consists of” (or variations thereof) appears in a clause of the body of a claim, rather than immediately following the preamble, it limits only the element set forth in that clause; other elements are not excluded from the claim as a whole. As used herein, the phrase “consisting essentially of” limits the scope of a claim to the specified elements or method steps, plus those that do not materially affect the basis and novel characteristic(s) of the claimed subject matter.

[0025] With respect to the terms “comprising”, “consisting of” and “consisting essentially of”, where one of these three terms are used herein, the presently disclosed and claimed subject matter may include the use of either of the other two terms. Thus, in some embodiments not otherwise explicitly recited, any instance of “comprising” may be replaced by “consisting of” or, alternatively, by “consisting essentially of”.

[0026] Other than in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein are to be understood as modified in all instances by the term “about”, having regard to normal tolerances in the art. The examples are not intended to limit the scope of the invention. In what follows, or where otherwise indicated, “%” will mean “weight %”, “ratio” will mean “weight ratio” and “parts” will mean “weight parts”.

[0027] The term “substantially” as used herein shall mean comprising more than 50%, where relevant, unless otherwise indicated.

[0028] The recitation of a numerical range using endpoints includes all numbers subsumed within that range (e.g., 1 to 5 includes 1, 1.5, 2, 2.75, 3, 3.80, 4, 5, etc.).

[0029] The terms “preferred” and “preferably” refer to embodiments of the invention that may afford certain benefits, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful and is not intended to exclude other embodiments from the scope of the invention.

[0030] It must also be noted that, as used in the specification and the appended claims, the singular forms “a”, “an” and “the” include plural referents unless the context clearly dictatesotherwise.

[0031] The person skilled in the art would appreciate that the embodiments described herein are exemplary only and that the electrical characteristics of the present application may be configured in a variety of alternative arrangements without departing from the spirit or the scope of the invention.

[0032] Although example embodiments of the disclosed technology are explained in detail herein, it is to be understood that other embodiments are contemplated. Accordingly, it is not intended that the disclosed technology be limited in its scope to the details of construction and arrangement of components set forth in the following description or illustrated in the drawings. The disclosed technology is capable of other embodiments and of being practised or carried out in various ways.

[0033] The term “heteroaryl,” as used herein, means an aromatic monocyclic ring or an aromatic bicyclic ring. The aromatic monocyclic rings are five or six membered rings containing 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of N, O, and S. The nitrogen heteroatoms can be optionally quaternised or oxidized to the N- oxide. The nitrogen containing rings can be optionally N-protected. The five membered aromatic monocyclic rings have two double bonds and the six membered aromatic monocyclic rings have three double bonds. The aromatic bicyclic rings are composed of an aromatic monocyclic ring fused to a phenyl group. Alternatively, aromatic bicyclic rings are composed of an aromatic monocyclic ring fused to another aromatic monocyclic ring. The aromatic monocyclic rings and the aromatic bicyclic rings are connected to the parent molecular moiety through a carbon or nitrogen atom. Representative examples of heteroaryl include, but are not limited to, benzothienyl, benzoxadiazolyl, cinnolinyl, dibenzofuranyl, furopyridinyl, furyl, imidazolyl, indazolyl, indolyl, isoxazolyl, isoquinolinyl, isothiazolyl, naphthyridinyl, oxadiazolyl, oxazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, pyrazolyl, pyridinium N-oxide, pyrrolyl, quinolinyl, tetrazolyl, thiadiazolyl, thiazolyl, thienopyridinyl, thienyl, triazolyl, and triazinyl.

[0034] The heteroaryl groups of the present invention are substituted with 0, 1, 2, 3, or 4 substituents independently selected from alkenyl, -ORa, alkylORa, -C(O)ORa, alkyl, - C(O)Ra, -OC(O)Ra, -SRa, alkynyl, -C(O)O- cyano, formyl, haloalkoxy, haloalkyl, halogen, hydroxy, hydroxyalkyl, mercapto, nitro, -NRcRc, and (NRcRd)carbonyl, wherein Rcand Rdare independently selected from hydrogen, alkyl, -C(O)Ra, formyl, aryl and arylalkyl. Representative examples include, but are not limited to, 3-cyanopyridin-2-yl, 5- hydroxypyridin-2-yl, and 3-methylpyridin-2-yl.

[0035] The term “heterocycle” as used herein, refers to a monocyclic or bicyclic, non- aromatic, saturated or partially unsaturated ring system. Monocyclic ring systems are exemplified by any 4-membered ring containing a heteroatom independently selected from oxygen, nitrogen and sulfur; or a 5-, 6-, 7-, or 8-membered ring containing one, two or three heteroatoms wherein the heteroatoms are independently selected from nitrogen, oxygen and sulfur. The 5-membered ring has 0 or 1 double bond. The 6-memebered ring has 0, 1 or 2 double bonds. The 7- or 8-membered ring has 0, 1, 2 or 3 double bonds.

[0036] Representative examples of monocyclic ring systems include, but are not limited to azetidinyl, azepanyl, azepinyl, diazepinyl, dioxolanyl, dioxanyl, dithianyl, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl, 3- oxo-morpholinyl, oxadiazolinyl, oxadiazolidinyl, oxazolinyl, 2-oxo-oxazolinyl, oxazolidinyl, piperazinyl, piperidyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuryl, tetrahydropyranyl, tetrahydropyridyl, tetrahydrothienyl, thiadiazolinyl, thiadiazolidinyl, thiazolinyl, thiazolidinyl, thiomorpholinyl, 1,1- dioxidothiomorpholinyl (thiomorpholine sulfone), thiopyranyl, 1,4-diazepanyl and trithianyl.

[0037] Bicyclic ring systems are exemplified by any of the above monocyclic ring systems fused to a phenyl group, a monocyclic cycloalkenyl group, as defined herein, a monocyclic cycloalkyl group, as defined herein, or an additional monocyclic heterocycle group, as defined herein.

[0038] Representative examples of bicyclic ring systems include but are not limited to, benzodioxinyl, benzopyranyl, benzothiopyranyl, 2,3-dihydroindolyl, indolizinyl, pyranopyridinyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, thiopyranopyridinyl, 2-oxo- 1,3-benzoxazolyl, 3-oxo-benzoxazinyl, 3-azabicyclo[3.2.0]heptyl, 3,6- diazabicyclo[3.2.0]heptyl, octahydrocyclopenta[c]pyrrolyl, hexahydro-1H-furo[3,4- c]pyrrolyl, and octahydropyrrolo[3,4-c]pyrrolyl. The monocyclic or bicyclic ring systems as defined herein may have two of the non-adjacent carbon atoms connected by a heteroatom selected from nitrogen, oxygen or sulfur, or an alkylene bridge between one and three additional carbon atoms.

[0039] Representative examples of monocyclic or bicyclic ring systems that contain such connection between two non-adjacent carbon atoms include, but not limited to, 2- azabicyclo[2.2.2]octyl, 2-oxa-5-azabicyclo[2.2.2]octyl, 2,5-diazabicyclo[2.2.2]octyl, 2- azabicyclo[2.2.1]heptyl, 2-oxa-5-azabicyclo[2.2.1]heptyl, 2,5-diazabicyclo[2.2.1]heptyl, 2- azabicyclo[2.1.1]hexyl, 5-azabicyclo[2.1.1]hexyl, 3-azabicyclo[3.1.1]heptyl, 6-oxa-3-azabicyclo[3.1.1]heptyl, 8-azabicyclo[3.2.1]octyl, 3-oxa-8-azabicyclo[3.2.1]octyl, 1,4- diazabicyclo[3.2.2]nonyl, 1,4-diazatricyclo[4.3.1.13,8]undecyl, 3,10- diazabicyclo[4.3.1]decyl, or 8-oxa-3-azabicyclo[3.2.1]octyl, octahydro-1H-4,7- methanoisoindolyl, and octahydro-1H-4,7-epoxyisoindolyl.

[0040] The heterocycle groups of this invention, including the representative examples listed above, can be optionally substituted with 1, 2, or 3 substituents independently selected from alkenyl, -ORa, -C(O)ORa, -alkylC(O)ORa, alkyl, -CORa, -OC(O)Ra, alkylOC(O)Ra, - S(O)Ra, -S(O)2Ra, -S(O)2NRaRb, alkynyl, -C(O)NRaRb, cyano, halo, haloalkyl, haloalkoxy, nitro, -NRaRb, and (NRaRb)alkyl, wherein Ra and Rb are independently selected from the group consisting of hydrogen, alkyl, aryl, haloalkyl and arylalkyl; furyl, imidazolyl, isothiazolyl, isoxazolyl, naphthyl, oxadiazolyl, oxazolyl, phenyl, pyrazinyl, pyrazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrrolyl, tetrazinyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazinyl, triazolyl, benzimidazolyl, benzothiazolyl, benzothiadiazolyl, benzothienyl, benzoxadiazolyl, benzoxazolyl, benzofuranyl, cinnolinyl, indolyl, naphthyridinyl, isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolinyl, and quinolinyl wherein said furyl, imidazolyl, isothiazolyl, isoxazolyl, naphthyl, oxadiazolyl, oxazolyl, phenyl, pyrazinyl, pyrazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrrolyl, tetrazinyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazinyl, triazolyl, benzimidazolyl, benzothiazolyl, benzothiadiazolyl, benzothienyl, benzoxadiazolyl, benzoxazolyl, benzofuranyl, cinnolinyl, indolyl, naphthyridinyl, isobenzofuranyl, isobenzothienyl, isoindolyl, isoquinolinyl, and quinolinyl may be substituted with 1 or 2 substituents independently selected from alkenyl, - ORa, -C(O)ORa, alkylC(O)ORa, alkyl, -CORa, -OC(O)Ra, alkylOC(O)Ra, -S(O)Ra, -SO2Ra, - SO2NRaRb, alkynyl, -C(O)NRaRb, cyano, halo, haloalkyl, haloalkoxy, nitro, -NRaRb, and (NRaRb)alkyl, wherein Raand Rbare independently selected from the group consisting of hydrogen, alkyl, aryl, haloalkyl and arylalkyl. The heterocycle groups of this invention are connected to the parent molecular moiety through any substitutable carbon or nitrogen atom in the groups. The nitrogen heteroatom may or may not be quaternised and may or may not be oxidized to the N-oxide. In addition, the nitrogen containing heterocyclic rings may or may not be N-protected.

[0041] In the context of this specification, the term “effective amount” includes a non-toxic but sufficient amount of an active compound to provide the stated / desired effect. Those skilled in the art will appreciate that the exact amount of a compound required will vary based on a number of factors and thus it is not possible to specify an exact “effective amount”. However, for any given case an appropriate “effective amount” may be determinedby one of ordinary skill in the art.

[0042] In the context of this specification, the term “therapeutically effective amount” includes a non-toxic but sufficient amount of an active compound to provide the desired therapeutic effect. Those skilled in the art will appreciate that the exact amount of a compound required will vary based on a number of factors and thus it is not possible to specify an exact “therapeutically effective amount”. However, for any given case an appropriate “therapeutically effective amount” may be determined by one of ordinary skill in the art.

[0043] In the context of this specification, the terms “treating” and “treatment” refer to any and all uses, which remedy the stated disease or symptoms thereof, hinder, retard or otherwise reverse the progression of the disease or other undesirable symptoms in any way whatsoever. Thus, the terms “treating” and “treatment” are to be considered in their broadest context. For example, treatment does not necessarily imply that a subject is treated until total recovery.

[0044] In the context of this specification the term “associated with” when used in the context of a disease or condition “associated with” the P2X7R means that the disease or condition, or a symptom thereof, may result from, be characterized by, or otherwise related to P2X7R. Thus, the association between the disease or condition and P2X7R activity may be direct or indirect and may be temporally separated.

[0045] In the context of this specification, the term “subject” includes human and also nonhuman animals. As such, in addition to being useful in the treatment of diseases and conditions in humans, the compounds of the present invention also find use in the treatment of diseases in non-human animals, for example mammals such as companion animals and farm animals. Non-limiting examples of companion animals and farm animals include dogs, cats, horses, cows, sheep and pigs. Preferably, the subject is a human.

[0046] 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 by any appropriate means. Summary of the Invention

[0047] The “PKT400” series has been synthesised as PKT401, PKT402 and PKT403.

[0048] The “PKT400” series has been synthesised and assessed for in vitro P2X7R antagonism in a functional dye uptake assay, and for antagonism of IL-1β and IL18 release (see, Table 4). The “PKT400” series has been assessed in an ex vivo patient Peripheral Blood Mononuclear Cell (PBMC) assay measuring IL-1β and IL18 release (see, Figure 1). The ”PKT400” series has been assessed for in vitro clearance in liver microsomal and hepatocyte studies (see, Table 5 and Table 6).

[0049] A PKT400 lead compound is planned to be assessed in the following in vivo models for CVD: Tandem stenosis; myocardial infarction; bleomycin pulmonary hypertension model; and monocrotaline pulmonary hypertension model. Table 1. Compounds of Formula (I) according to the present invention

[0050] According to a first aspect of the present invention there is provided a compound of Formula (I):(I)

[0051] wherein are or

[0052] with the proviso that R1, R2and R3cannot each be H;

[0053] or a pharmaceutically-acceptable salt thereof.

[0054] In an embodiment, R1, R2 and R3 are each F. This compound is designated “PKT401”.

[0055] In an embodiment, R1and R2are F and R3is H. This compound is designated “PKT402”.

[0056] In an embodiment, R1is F and R2and R3are H. This compound is designated “PKT403”.

[0057] In an embodiment, the pharmaceutically-acceptable salt is a hydrochloride salt.

[0058] In an embodiment, the compound is a P2X7 receptor antagonist.

[0059] According to a second aspect of the present invention there is provided a pharmaceutical composition comprising a compound of Formula (I) according to the first aspect of the present invention or a pharmaceutically-acceptable salt thereof, together with a pharmaceutically acceptable carrier, diluent or excipient.

[0060] According to a third aspect of the present invention there is provided a method for the treatment of a disease or condition associated with P2X7R in a subject in need thereof, the method comprising administration to the subject of a therapeutically effective amount of a compound according to the first aspect of the present invention, a pharmaceutically- acceptable salt thereof, or a composition according to the second aspect of the present invention.

[0061] In an embodiment, the disease or condition is associated with upregulation or increased expression of P2X7R.

[0062] In an embodiment, the disease or condition is associated with activation of P2X7R.

[0063] In an embodiment, the disease or condition is a neuroinflammatory disease or a neurodegenerative disease.

[0064] In an embodiment, the disease or condition is a disease of the central nervous system.

[0065] In an embodiment, the disease or condition is selected from the group consisting of:pain, rheumatoid arthritis, osteoarthritis, sepsis, psoriasis, allergic dermatitis, asthma, chronic obstructive pulmonary disease, airways hyper-responsiveness, septic shock, epilepsy, glomerulonephritis, irritable bowel syndrome, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, coronary artery disease, cardiovascular disease, acute coronary syndrome, myocarditis, pericarditis, atherosclerosis, myocardial ischemia, reperfusion injury, cancer, myeloblastic leukemia, diabetes, Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, Huntington’s disease, glaucoma, multiple sclerosis, amyotrophic lateral sclerosis, depression, age-related macular degeneration, uveitis, neuropathic pain, depression, bipolar affective disorders, anxiety, meningitis, traumatic brain injury, acute spinal cord injury, neuropathic pain, osteoporosis, burn injury, ischemic heart disease, myocardial infarction, stroke, varicose veins, tandem stenosis, bleomycin pulmonary hypertension and monocrotaline pulmonary hypertension.

[0066] In an embodiment, the disease or condition is selected from the group consisting of: cardiovascular disease, atherosclerosis, myocardial infarction, tandem stenosis, bleomycin pulmonary hypertension and monocrotaline pulmonary hypertension.

[0067] In an embodiment, the disease or condition is cardiovascular disease.

[0068] According to a fourth aspect of the present invention there is provided use of a compound according to the first aspect of the present invention, a pharmaceutically- acceptable salt thereof, or a composition according to the second aspect of the present invention, in the manufacture of a medicament for the treatment or prevention of a disease or condition associated with P2X7R in a subject.

[0069] In an embodiment, the disease or condition is associated with upregulation or increased expression of P2X7R.

[0070] In an embodiment, the disease or condition is associated with activation of P2X7R.

[0071] In an embodiment, the disease or condition is a neuroinflammatory disease or a neurodegenerative disease.

[0072] In an embodiment, the disease or condition is a disease of the central nervous system.

[0073] In an embodiment, the disease or condition is selected from the group consisting of: pain, rheumatoid arthritis, osteoarthritis, sepsis, psoriasis, allergic dermatitis, asthma, chronic obstructive pulmonary disease, airways hyper-responsiveness, septic shock, epilepsy, glomerulonephritis, irritable bowel syndrome, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, coronary artery disease, cardiovascular disease, acute coronary syndrome, myocarditis, pericarditis, atherosclerosis, myocardial ischemia,reperfusion injury, cancer, myeloblastic leukemia, diabetes, Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, Huntington’s disease, glaucoma, multiple sclerosis, amyotrophic lateral sclerosis, depression, age-related macular degeneration, uveitis, neuropathic pain, depression, bipolar affective disorders, anxiety, meningitis, traumatic brain injury, acute spinal cord injury, neuropathic pain, osteoporosis, burn injury, ischemic heart disease, myocardial infarction, stroke, varicose veins, tandem stenosis, bleomycin pulmonary hypertension and monocrotaline pulmonary hypertension.

[0074] In an embodiment, the disease or condition is selected from the group consisting of: cardiovascular disease, atherosclerosis, myocardial infarction, tandem stenosis, bleomycin pulmonary hypertension and monocrotaline pulmonary hypertension.

[0075] In an embodiment, the disease or condition is cardiovascular disease.

[0076] According to a fifth aspect of the present invention there is provided a compound according to the first aspect of the present invention, a pharmaceutically-acceptable salt thereof, or a composition according to the second aspect of the present invention, for use in the treatment or prevention of a disease or condition selected from the group consisting of: pain, rheumatoid arthritis, osteoarthritis, sepsis, psoriasis, allergic dermatitis, asthma, chronic obstructive pulmonary disease, airways hyper-responsiveness, septic shock, epilepsy, glomerulonephritis, irritable bowel syndrome, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, coronary artery disease, cardiovascular disease, acute coronary syndrome, myocarditis, pericarditis, atherosclerosis, myocardial ischemia, reperfusion injury, cancer, myeloblastic leukemia, diabetes, Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, Huntington’s disease, glaucoma, multiple sclerosis, amyotrophic lateral sclerosis, depression, age-related macular degeneration, uveitis, neuropathic pain, depression, bipolar affective disorders, anxiety, meningitis, traumatic brain injury, acute spinal cord injury, neuropathic pain, osteoporosis, burn injury, ischemic heart disease, myocardial infarction, stroke, varicose veins, tandem stenosis, bleomycin pulmonary hypertension and monocrotaline pulmonary hypertension.

[0077] According to a sixth aspect of the present invention there is provided a method for the synthesis of a compound according to claim 2, the method comprising the steps of:

[0078] a) forming (3s,5s,7s)-3,5,7-trifluoroadamantane-1-carboxamide from the reaction of (3s,5s,7s)-3,5,7-trifluoroadamantane-1-carboxylic acid and 1,1’-carbonyldiimidazole;

[0079] b) reducing the (3s,5s,7s)-3,5,7-trifluoroadamantane-1-carboxamide with lithium aluminium hydride to give ((3s,5s,7s)-3,5,7-trifluoroadamantan-1-yl)methanamine hydrochloride;

[0080] c) reacting the ((3s,5s,7s)-3,5,7-trifluoroadamantan-1-yl)methanamine hydrochloride with 2-chloro-5-iodobenzoic acid, oxalyl chloride, DMF and then triethylamine to give 2- chloro-5-iodo-N-(((3s,5s,7s)-3,5,7-trifluoroadamantan-1-yl)methyl)benzamide;

[0081] d) reacting the 2-chloro-5-iodo-N-(((3s,5s,7s)-3,5,7-trifluoroadamantan-1- yl)methyl)benzamide with tetrabutylammonium chloride, diisopropylethylamine, Pd(II)OAc and toluene to provide 2-chloro-5-(3-oxopropyl)-N-(((3s,5s,7s)-3,5,7-trifluoroadamantan-1- yl)methyl)benzamide; and

[0082] e) reacting the 2-chloro-5-(3-oxopropyl)-N-(((3s,5s,7s)-3,5,7-trifluoroadamantan-1- yl)methyl)benzamide with aminopropanol, acetic acid and then sodium cyanoborohydride to give 2-chloro-5-(3-((3-hydroxypropyl)amino)propyl)-N-(((3s,5s,7s)-3,5,7- trifluoroadamantan-1-yl)methyl)benzamide hydrochloride.

[0083] According to another aspect of the present invention there is provided a pharmaceutical composition comprising a compound of Formula (I) according to the first aspect together with a pharmaceutically acceptable carrier, diluent or excipient.

[0084] In various embodiments, the disease or condition may be associated with upregulation or increased expression of P2X7R.

[0085] In other embodiments, the disease or condition may be associated with activation of the P2X7R.

[0086] In other embodiments, the disease or condition may be an inflammatory disease or condition. In one embodiment, the disease or condition is a neuroinflammatory disease or a neurodegenerative disease.

[0087] In other embodiments, the disease or condition may be a disease of the central nervous system.

[0088] In some embodiments the disease or condition may be one or more of; pain, rheumatoid arthritis, osteoarthritis, sepsis, psoriasis, allergic dermatitis, asthma, chronic obstructive pulmonary disease, airways hyper-responsiveness, septic shock, epilepsy, glomerulonephritis, irritable bowel syndrome, inflammatory bowel disease, Crohn's disease, ulcerative colitis, coronary artery disease, cardiovascular disease, acute coronary syndrome, myocarditis, pericarditis, atherosclerosis, myocardial ischemia, reperfusion injury, cancer, myoblastic leukaemia, diabetes, Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, Huntington’s disease, glaucoma, multiple sclerosis, amyotrophic lateral sclerosis, depression, age-related macular degeneration, uveitis, neuropathic pain, depression, bipolar affective disorders, anxiety, meningitis, traumatic brain injury, acute spinal cord injury, osteoporosis, burn injury, ischemic heart disease, myocardial infarction, stroke and varicoseveins.

[0089] In one embodiment the disease or condition is coronary artery disease, cardiovascular disease or atherosclerosis.

[0090] In another embodiment the disease or condition is coronary artery disease.

[0091] In still a further embodiment the disease or condition is acute coronary syndrome, coronary artery disease, myocarditis, pericarditis, myocardial ischemia and reperfusion injury.

[0092] In another aspect, the present invention provides a method for modulating P2X7R activity in a subject in need thereof, the method comprising administration to the subject of an effective amount of a compound of Formula (I) according to the first aspect.

[0093] Modulating P2X7R activity may involve inhibiting P2X7R activity.

[0094] Compounds and pharmaceutical compositions of the present invention may be administered via any route that delivers an effective amount of the compounds to the tissue or site to be treated. In general, the compounds and compositions may be administered by the parenteral (for example intravenous, intraspinal, subcutaneous or intramuscular), oral, inhalation, or topical route. Administration may be systemic, regional or local.

[0095] The particular route of administration to be used in any given circumstance will depend on a number of factors, including the nature of the disease or condition to be treated, the severity and extent of the disease or condition, the required dosage of the particular compound to be delivered and the potential side-effects of the compound.

[0096] In general, suitable compositions may be prepared according to methods that are known to those of ordinary skill in the art and may include pharmaceutically acceptable carriers, diluents and / or excipients. The carriers, diluents and excipients must be “acceptable” in terms of being compatible with the other ingredients of the composition, and not deleterious to the recipient thereof.

[0097] Examples of pharmaceutically acceptable carriers or diluents are demineralised or distilled water; saline solution; vegetable-based oils such as peanut oil, safflower oil, olive oil, cottonseed oil, maize oil or coconut oil; silicone oils, including polysiloxanes, such as methyl polysiloxane, phenyl polysiloxane and methylphenyl polysiloxane; volatile silicones; mineral oils such as liquid paraffin, soft paraffin or squalane; cellulose derivatives such as methyl cellulose, ethyl cellulose, carboxymethylcellulose, sodium carboxymethylcellulose or hydroxypropylmethylcellulose; Cremophor; cyclodextrins; lower alcohols, for example ethanol or / -propanol; lower polyalkylene glycols or lower alkylene glycols, for example polyethylene glycol, polypropylene glycol, ethylene glycol, propylene glycol, 1 ,3-butyleneglycol or glycerin; fatty acid esters such as isopropyl palmitate, isopropyl myristate or ethyl oleate; polyvinylpyrridone; agar; carrageenan; gum tragacanth or gum acacia and petroleum jelly. Typically, the carrier or carriers will form from 10% to 99.9% by weight of the compositions.

[0098] Pharmaceutical compositions of the invention may be in a form suitable for administration by injection, in the form of a formulation suitable for oral ingestion (such as capsules, tablets, caplets and elixirs for example), in the form of an ointment, cream or lotion suitable for topical administration, in a form suitable for delivery as an eye drop, in an aerosol form suitable for administration by inhalation, such as by intranasal inhalation or oral inhalation, in a form suitable for parenteral administration, that is, subcutaneous, intramuscular or intravenous injection.

[0099] For administration as an injectable solution or suspension, non-toxic parenterally acceptable diluents or carriers can include cyclodextrins (for example Captisol®) Cremaphor, Ringer’s solution, isotonic saline, phosphate buffered saline, ethanol and 1,2- propylene glycol. To aid injection and delivery, the compounds may also be added to PEG and non-PEGylated liposomes or micelles with specific targeting tags attached to PEG moieties, such as the RGD peptide or glutathione, for aiding passage across the blood brain barrier.

[0100] Some examples of suitable carriers, diluents, excipients and adjuvants for oral use include cyclodextrins, Cremophor, peanut oil, liquid paraffin, sodium carboxymethyl cellulose, methylcellulose, sodium alginate, gum acacia, gum tragacanth, dextrose, sucrose, sorbitol, mannitol, gelatine and lecithin. In addition, these oral formulations may contain suitable flavouring and colourings agents. When used in capsule form the capsules may be coated with compounds such as glyceryl monostearate or glyceryl distearate that delay disintegration.

[0101] Adjuvants typically include emollients, emulsifiers, thickening agents, preservatives, bactericides and buffering agents.

[0102] Solid forms for oral administration may contain binders acceptable in human and veterinary pharmaceutical practice, sweeteners, disintegrating agents, diluents, flavourings, coating agents, preservatives, lubricants and / or time delay agents. Suitable binders include gum acacia, gelatine, corn starch, gum tragacanth, sodium alginate, carboxymethylcellulose or polyethylene glycol. Suitable sweeteners include sucrose, lactose, glucose, aspartame or saccharin. Suitable disintegrating agents include corn starch, methylcellulose, polyvinylpyrrolidone, guar gum, xanthan gum, bentonite, alginic acid or agar. Suitablediluents include lactose, sorbitol, mannitol, dextrose, kaolin, cellulose, calcium carbonate, calcium silicate or dicalcium phosphate. Suitable flavouring agents include peppermint oil, oil of wintergreen, cherry, orange or raspberry flavouring. Suitable coating agents include polymers or copolymers of acrylic acid and / or methacrylic acid and / or their esters, waxes, fatty alcohols, zein, shellac or gluten. Suitable preservatives include sodium benzoate, vitamin E, alpha-tocopherol, ascorbic acid, methyl paraben, propyl paraben or sodium bisulphite. Suitable lubricants include magnesium stearate, stearic acid, sodium oleate, sodium chloride or talc. Suitable time- delay agents include glyceryl monostearate or glyceryl distearate.

[0103] Liquid forms for oral administration may contain, in addition to the above agents, a liquid carrier. Suitable liquid carriers include water, oils such as olive oil, peanut oil, sesame oil, sunflower oil, safflower oil, coconut oil, liquid paraffin, ethylene glycol, propylene glycol, polyethylene glycol, ethanol, propanol, isopropanol, glycerol, fatty alcohols, triglycerides or mixtures thereof.

[0104] Suspensions for oral administration may further comprise dispersing agents and / or suspending agents. Suitable suspending agents include sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, sodium alginate or acetyl alcohol. Suitable dispersing agents include lecithin, polyoxyethylene esters of fatty acids, such as stearic acid, polyoxyethylene sorbitol mono- or di-oleate, -stearate or -laurate, polyoxyethylene sorbitan mono- or di-oleate, -stearate or -laurate and the like.

[0105] Emulsions for oral administration may further comprise one or more emulsifying agents. Suitable emulsifying agents include dispersing agents as exemplified above or natural gums such as guar gum, gum acacia or gum tragacanth.

[0106] A further suitable emulsifying agent for use in oral or parenteral formulations, which may also function as a solubilizer, is Kolliphor® HS 15.

[0107] Methods for preparing parenterally administrable compositions are apparent to those skilled in the art, and are described in more detail in, for example, Remington’s Pharmaceutical Science, 15th ed., Mack Publishing Company, Easton, PA, hereby incorporated by reference herein.

[0108] Topical formulations may comprise an active ingredient together with one or more acceptable carriers, and optionally any other therapeutic ingredients. Formulations suitable for topical administration include liquid or semi-liquid preparations suitable for penetration through the skin to the site where treatment is required, such as liniments, lotions, creams, ointments or pastes, and drops suitable for administration to the eye, ear or nose.

[0109] Drops according to the present invention may comprise sterile aqueous or oily solutions or suspensions. These may be prepared by dissolving the active ingredient in an aqueous solution of a bactericidal and / or fungicidal agent and / or any other suitable preservative, and optionally including a surface active agent. The resulting solution may then be clarified by filtration, transferred to a suitable container and sterilised. Sterilisation may be achieved by autoclaving or maintaining at 90 °C to 100 °C for half an hour, or by filtration, followed by transfer to a container by an aseptic technique. Examples of bactericidal and fungicidal agents suitable for inclusion in the drops are phenylmercuric nitrate or acetate (0.002%), benzalkonium chloride (0.01 %) and chlorhexidine acetate (0.01 %). Suitable solvents for the preparation of an oily solution include glycerol, diluted alcohol and propylene glycol.

[0110] Lotions according to the present invention include those suitable for application to the skin or eye. An eye lotion may comprise a sterile aqueous solution optionally containing a bactericide and may be prepared by methods similar to those described above in relation to the preparation of drops. Lotions or liniments for application to the skin may also include an agent to hasten drying and to cool the skin, such as an alcohol or acetone, and / or a moisturiser such as glycerol, or oil such as olive oil.

[0111] Creams, ointments or pastes according to the present invention are semi-solid formulations of the active ingredient for external application. They may be made by mixing the active ingredient in finely divided or powdered form, alone or in solution or suspension in an aqueous or non-aqueous fluid, with a greasy or non-greasy basis. The basis may comprise hydrocarbons such as hard, soft or liquid paraffin, glycerol, beeswax, a metallic soap; a mucilage; an oil of natural origin such as almond, corn, arachis, castor or olive oil; wool fat or its derivatives, or a fatty acid such as stearic or oleic acid together with an alcohol, such as propylene glycol or macrogols.

[0112] The composition may incorporate any suitable surfactant such as an anionic, cationic or non-ionic surfactant, such as sorbitan esters or polyoxyethylene derivatives thereof. Suspending agents such as natural gums, cellulose derivatives or inorganic materials such as silicaceous silicas, and other ingredients such as lanolin, may also be included.

[0113] In some embodiments the compositions are administered in the form of suppositories suitable for rectal administration of the compounds. These compositions are prepared by mixing the compound with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the zeolite or zeolite-like material. Such materials include cocoa butter, glycerinatedgelatin, hydrogenated vegetable oils, mixtures of polyethylene glycols of various molecular weights and fatty acid esters of polyethylene glycol.

[0114] The compositions may also be administered or delivered to target cells in the form of liposomes. Liposomes are generally derived from phospholipids or other lipid substances and are formed by mono- or multi-lamellar hydrated liquid crystals that are dispersed in an aqueous medium. Specific examples of liposomes used in administering or delivering a composition to target cells are synthetic cholesterol (Sigma), the phospholipid 1 ,2- distearoyl-sn-glycero-3-phosphocholine (DSPC); Avanti Polar Lipids), the PEG lipid 3-N- [(-methoxy polyethylene glycol)2000)carbamoyl]-1 ,2- dimyrestyloxy-propylamine (PEG- cDMA), and the cationic lipid 1 ,2-di-o-octadecenyl-3- (N,N-dimethyl)aminopropane (DODMA) or 1 ,2-dilinoleyloxy-3-(N,N- dimethyl)aminopropane (DLinDMA) in the molar ratios 55:20:10:15 or 48:20:2:30, respectively, PEG-cDMA, DODMA and DLinDMA. Any non-toxic, physiologically acceptable and metabolisable lipid capable of forming liposomes can be used. The compositions in liposome form may contain stablisers, preservatives, excipients and the like. The preferred lipids are the phospholipids and the phosphatidyl cholines (lecithins), both natural and synthetic. Methods to form liposomes are known in the art, and in relation to this, specific reference is made to: Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic Press, New York, N.Y. (1976), p.33 et seq., the contents of which is incorporated herein by reference.

[0115] The compositions may also be administered in the form of microparticles or nanoparticles. Biodegradable microparticles formed from polyactide (PLA), polylactide- co- glycolide (PLGA), and epsilon-caprolactone (έ-caprlactone) have been extensively used as drug carriers to increase plasma half life and thereby prolong efficacy (R. Kumar, M., 2000, J. Pharm. Pharmaceut. Sci.3(2) 234-258). Microparticles have been formulated for the delivery of a range of drug candidates including vaccines, antibiotics, and DNA. Moreover, these formulations have been developed for various delivery routes including parenteral subcutaneous injection, intravenous injection and inhalation.

[0116] The compositions may incorporate a controlled release matrix that is composed of sucrose acetate isobutyrate (SAIB) and an organic solvent or organic solvents mixture. Polymer additives may be added to the vehicle as a release modifier to further increase the viscosity and slow down the release rate. SAIB is a well known food additive. It is a very hydrophobic, fully esterified sucrose derivative, at a nominal ratio of six isobutyrate to two acetate groups. As a mixed ester, SAIB does not crystallise but exists as a clear viscous liquid. Mixing SAIB with a pharmaceutically acceptable organic solvent, such as ethanol orbenzyl alcohol decreases the viscosity of the mixture sufficiently to allow for injection. An active pharmaceutical ingredient may be added to the SAIB delivery vehicle to form SAIB solution or suspension formulations. When the formulation is injected subcutaneously, the solvent differs from the matrix allowing the SAIB-drug or SAIB-drug-polymer mixtures to set up as an in situ forming depot.

[0117] For the purposes of the present invention compounds and compositions may be administered to subjects either therapeutically or preventively. In a therapeutic application compositions are administered to a patient already suffering from a disease or condition in an amount sufficient to cure or at least partially arrest the disease or condition and its complications. The composition should provide a quantity of the compound or agent sufficient to effectively treat the subject.

[0118] The therapeutically effective amount for any particular subject will depend upon a variety of factors including: the disease or condition being treated and the severity thereof; the activity of the compound administered; the composition in which the compound is present; the age, body weight, general health, sex and diet of the subject; the time of administration; the route of administration; the rate of sequestration of the compound; the duration of the treatment; drugs used in combination or coincidental with the compound, together with other related factors well known in medicine. One skilled in the art would be able, by routine experimentation, to determine an effective, non-toxic amount of a compound that would be required to treat or prevent a particular disease or condition.

[0119] Generally, an effective dosage is expected to be in the range of about 0.0001 mg to about 1000 mg per kg body weight per 24 hours; typically, about 0.001 mg to about 750 mg per kg body weight per 24 hours; about 0.01 mg to about 500 mg per kg body weight per 24 hours; about 0.1 mg to about 500 mg per kg body weight per 24 hours; about 0.1 mg to about 250 mg per kg body weight per 24 hours; about 1.0 mg to about 250 mg per kg body weight per 24 hours. More typically, an effective dose range is expected to be in the range about 1.0 mg to about 200 mg per kg body weight per 24 hours; about 1.0 mg to about 100 mg per kg body weight per 24 hours; about 1.0 mg to about 50 mg per kg body weight per 24 hours; about 1.0 mg to about 25 mg per kg body weight per 24 hours; about 5.0 mg to about 50 mg per kg body weight per 24 hours; about 5.0 mg to about 20 mg per kg body weight per 24 hours; about 5.0 mg to about 15 mg per kg body weight per 24 hours.

[0120] Alternatively, an effective dosage may be up to about 500 mg / m2. Generally, an effective dosage is expected to be in the range of about 25 to about 500 mg / m2, preferably about 25 to about 350 mg / m2, more preferably about 25 to about 300 mg / m2, still morepreferably about 25 to about 250 mg / m2, even more preferably about 50 to about 250 mg / m2, and still even more preferably about 75 to about 150 mg / m2.

[0121] Typically, in therapeutic applications, the treatment would be for the duration of the disease or condition.

[0122] Further, it will be apparent to one of ordinary skill in the art that the optimal quantity and spacing of individual dosages will be determined by the nature and extent of the disease or condition being treated, the form, route and site of administration, and the nature of the particular individual being treated. Also, such optimum conditions can be determined by conventional techniques. Brief Description of the Drawings

[0123] A preferred embodiment of the present invention will now be described having regard to the accompanying drawings / s in which:

[0124] Figure 1 shows adamantane amide P2X7R antagonists reduce BzATP-induced IL-1β and IL-18 from patient-derived monocytes. A) Measured as the relative reduction in IL-1β release. B) Measured as the relative reduction in IL-18 release. In particular, Figure 1, when read in conjunction with Table 2, demonstrates in vitro P2X7R antagonism data for AZD9056 (R1 = R2 = R3 = H). In particular, Figure 1 depicts IL-1β and IL-18 release from PBMC isolated from patients with no CAD (CACS = 0 Agatston) vs CAD (CACS > 0 Agatston) vs STEMI.

[0125] For comparative purposes, Table 3 demonstrates in vitro liver microsomal stability for AZD9056 (R1 = R2 = R3 = H).

[0126] Figure 1, read in conjunction with Table 4, demonstrates in vitro P2X7R antagonism data for “PKT400” series. In particular, Figure 1 depicts IL-1β and IL-18 release from PBMC isolated from patients with no CAD (CACS = 0 Agatston) vs CAD (CACS > 0 Agatston) vs STEMI.

[0127] Table 5 demonstrates in vitro liver microsomal stability for “PKT400” series.

[0128] Table 6 demonstrates in vitro hepatocyte stability for “PKT400” series. Methods and Materials – Pharmacological Studies THP-1 cell line

[0129] Tamm-Horsfall protein 1 (THP-1) cells are a human monocytic leukemia cell line derived from a 1-year-old male infant, and used extensively as a model of monocyte and macrophage function. Cells were cultured in suspension in T175 flasks (Corning), usingRPMI 1640 media (with ATCC modification; Gibco) supplemented with 10% heat- inactivated foetal bovine serum (FBS; Gibco). THP-1 cells were separated from old media by centrifugation (125 g, 5 min) and a proportion of cell pellet resuspended back into a T175 flask with fresh media. Dye uptake assay with THP-1 cells

[0130] THP-1 cells were harvested from culture by centrifugation (125 g, 5 min) and resuspended in RPMI 1640 media (Gibco) supplemented with 5% FBS (Gibco), 100 ng mL-1lipopolysaccharide (LPS; from E. coli strain 0111:B4, Sigma-Aldrich), and 10 ng mL-1recombinant human interferon gamma (IFN-γ; R&D Systems). Cells were immediately seeded onto 96-well black-walled, CellBIND plates (Corning) at a density of 1.5 x 105cells per well and incubated for 24 h (37oC, 5% CO2) to allow differentiation into macrophages. Supernatant was removed from each well, and cells were washed once with 150 μL of warm modified Hanks’ Balanced Salt Solution containing low Ca2+(HBSS; KCl 5.37 mM, KH2PO40.44 mM, NaCl 136.89 mM, Na2HPO40.34 mM, glucose 5.55 mM, NaHCO34.17 mM, CaCl20.1 mM, pH 7.4, 37oC).

[0131] Cells were treated with 100 μL of warmed HBSS buffer (as above, pH 7.4, 37oC) containing: (i) YO-PRO-1® iodide (1 μM; Life Technologies), (ii) 100 μM BzATP (Sigma- Aldrich) or vehicle control (0.4% ultra-pure water) (v / v), and (iii) test compound (1 nM – 10 μM) or vehicle control (0.1% DMSO) (v / v). Fluorescence in each well was immediately recorded every 30 s for 1.5 h at 37oC using a BMG POLARstar Omega (λex= 485-12, λem= 520).

[0132] Fluorescence values recorded at 1 h were used to determine the IC50 in GraphPad Prism 7.0 (GraphPad software, USA) by using a 4-paramater sigmoidal dose-response (inhibition) fit. Each test compound was assayed in duplicates and repeated a minimum of three times, with the calculated IC50 from each assay used to determine the average and standard error of the mean (SEM). Measurement of IL-1β release by THP-1 cells

[0133] Cultured THP-1 cells were harvested by centrifugation (125 g, 5 min), and resuspended in RPMI 1640 media (Gibco) supplemented with 25 ng mL-1LPS (E. coli strain 0111:B4, Sigma-Aldrich) and 10 ng mL-1IFN-γ (R&D Systems). Cells were seeded onto 96-well black-walled, CellBIND plates (Corning) at a density of 2 x 105cells per well, and incubated for 3 h (37oC, 5% CO2).

[0134] Supernatant was removed and cells incubated with test compound (0.1 – 10 μM) or vehicle control (0.1% DMSO) (v / v) diluted in RPMI 1640 media for 30 min at 37oC. Pre- treatments were aspirated, and cells were washed once with 150 μL dPBS buffer (pH 7.4, 37oC). Cells were treated with (i) 1 mM BzATP (Sigma-Aldrich) or vehicle control (0.4% ultra-pure water) (v / v), and (ii) test compound (0.1-10 μM) or vehicle control (0.1% DMSO) (v / v) for 1 h (37oC, 5% CO2). Supernatant was aspirated and centrifuged at 10,000 g for 10 s. The final supernatant was collected and stored at -80oC. Supernatant samples were defrosted and assayed for IL-1β with a human IL-1β ELISA kit (R&D Systems) as per the manufacturer’s instructions.

[0135] Absorbance was recorded using a BMG POLARstar Omega microplate reader set to 450 nm. Absorbance readings were basal corrected, and IL-1β concentration in each supernatant was determined by interpolating from a standard curve generated using a sigmoidal dose-response (variable slope) fit in GraphPad Prism 7.0 (GraphPad software, USA). Interpolated values were then expressed as a percentage of the positive control, and analysed using a 4-parameter sigmoidal dose-response (inhibition) fit to determine the IC50 of each test compound. Samples and standards were assayed in duplicates and repeated a minimum of three repetitions to calculate the average IC50and standard error of the mean. Measurement of IL-18 release by THP-1 cells

[0136] Once flasks were estimated to have reached between 80-100% confluency, THP-1 cells were aspirated into a 20 mL V-bottom falcon tube and centrifuged at 125 g for 5 minutes at room temperature with break on. The supernatant was removed, and the pellet was homogenised before being resuspended in 10 mL of RPMI-1640 [ATCC modified media, supplemented with low sodium bicarbonate, L-glutamine, sodium pyruvate, HEPES, phenol red and high glucose] (Gibco, USA).

[0137] A small proportion of cells were aspirated and diluted 1:1 with 0.4% trypan blue solution (Gibco, USA) and counted with a hemocytometer. The remaining cells are centrifuged at 125 g for 5 minutes at room temperature with break on and the supernatant is aspirated. THP-1 cells were resuspended in RMPI-1640 [ATCC modified media], 10 ng / mL recombinant human IFN-γ [carrier free, R&D systems] and 25 ng / mL LPS [from E. coli strain 0111: B4, Sigma-Aldrich] at a density of 1 x 106cells per 100 μL.

[0138] Treated THP-1 cells were seeded in a 96-well, Corning cellBIND® flat bottom clear microplates [Corning, USA) at a density of 2 x 105cells per well at a volume of 100 μL and incubated for 3-hours at 37oC in enriched 5% CO2environment. After this time, supernatantwas removed from the wells, and cells were pre-treated with antagonists resuspended at a concentration between 0.001-1 μΜ or 0.01% DMSO resuspended in 100 μL of RPM-1640 [ATCC modified media] for 30-minutes at 37oC in a 5% CO2 enriched environment. Pre- treatment was then aspirated, and cells were gently washed with 200 μL of phosphate- buffered saline (dPBS) without calcium or magnesium [pH range between 7.0-7.3, osmolarity 270 – 300 mOsm / kg, Gibco, USA].

[0139] Cells were then treated with either (i) 0.01% DMSO, (ii) 1 mM BzATP only or (iii) 1 mM BzATP in combination with P2X7 receptor antagonists at a concentration of 0.001 - 1 μΜ in dPBS without calcium or magnesium for 1 hour at 37oC in a 5% enriched CO2 environment. The supernatant was collected into a clean RNAase and DNAase free Eppendorf and centrifuged at 125 g for 5 minutes at room temperature. Supernatant was then transferred into a new clean Eppendorf tube to eliminate potential contamination from cells. Eppendorf tubes were stored in -80oC for downstream analysis.

[0140] Total IL-18 capture antibodies (commonly name that identifies a monoclonal antibody) were plated in 96-well clear, flat bottom polystyrene microplates with high-bind treatment (R & D Systems, United States) overnight at room temperature. Plates that were not used immediately the next day were stored at 4oC, sealed for up to a week. Before sample preparation, plates were washed with 0.05% Tween® 20 in PBS, pH 7.2-7.4 diluted in Ultrapure water 3-times, removing solution between each wash. Wells were then blocked with 0.1% BSA in PBS, diluted in Ultrapure water for a minimum of 1-hour at room temperature.

[0141] Samples for IL-18 detection were prepared at a 1:1 ratio in 0.1% BSA and incubated in the wells for 2-hours at room temperature to achieve adequate capture. After this, samples were removed, and wells were washed with 0.05% Tween® 20 as detailed above. A second antibody aimed to detect the attached protein (often referred to as the “detection” antibody and typically a high-affinity monoclonal antibody) was incubated for 2-hours at room temperature. The plate was washed as detailed above and 1:40 dilution of streptavidin conjugated with horseradish-peroxidase enzyme (Strep-HRP) was diluted in 0.1% BSA and added to the plate for 20-minutes at room temperature.

[0142] Again, the plate was washed to remove unbound Strep-HRP, and a 1:1 mixture of colour reagent A (known as H2O2) and colour reagent B (Tetramethylbenzidine) was reacting with HRP to induce a colorimetric change from clear to dark blue (colour depends on the amount of cytokine detected) at room temperature. The reaction was stopped after 20- minutes with stop solution (2N H2SO4) and was read on a microplate set to 450 nm, with awavelength correction set to 540 nm. Peripheral Blood Mononuclear Cell (PBMC) Isolation

[0143] PBMCs were isolated from a cohort of 70 participants recruited to the BioHEART study (ACTRN12618001322224), undergoing clinically indicated CT angiography.9 mL of blood was collected from venous cannulation into a vacuette lithium heparin tube (Greiner Vacuette, Kremsmunster, Austria). Whole blood was diluted 1:1 with Hanks’ Balanced Salt Solution (HBSS) with calcium chloride (CaCl2) and magnesium chloride (MgCl2) (Thermo Fisher Scientific, Massachusetts, US).13 mL of Ficoll-Paque media was added to a 50 mL Falcon tube and the diluted blood sample was added carefully on top of the Ficoll-Paque media, density 1.077 g / mL (Cytiva) and centrifuged at 2000 rpm for 15-minutes at room temperature with break off. PBMCs were collected from the interphase region and washed with 8 mL of HBSS with CaCl2 and MgCl2 via gentle inversion and centrifuged at 2000 rpm for 10-minutes at room temperature with break off. The PBMCs form a pellet with various levels of contamination from erythrocytes.

[0144] For the purpose of downstream activities, PBMCs isolated from homolysed samples were treated with 2 mL of 0.8% ammonium chloride diluted in 0.1 mM EDTA for 2-minutes to aid the lysis of erythrocytes (Stem Cell Technologies, Vancouver, Canada) and then resuspended with 8 mL of HBSS with CaCl2 and MgCl2.

[0145] For samples with minimal erythrocyte contamination, supernatant was removed from these samples and the PBMCs were washed a second time via resuspension in 10 mL of HBSS with CaCl2 and MgCl2. Before repeating the above centrifugation procedure, 10 μL of resuspended PBMCs were removed and diluted 1:1 with trypan blue for counting. Supernatant was removed and PBMCs were resuspended at 10 x 106cells / mL in RPMI-1640 with 2 mM L-Glutamine but without heat inactivated bovine serum (HIBS) (Thermo Fisher Scientific, Massachusetts, US).

[0146] PBMCs were plated in duplicate in a 96-well flat bottom CellBIND® microplate (Corning, New York, USA) at a density of 1.6 x 105cells / well in 100 μL of RMPI-1640 as listed above and incubated for 3-hours at 37 ^c in a 5% CO2enriched environment. PBMC Stimulation

[0147] Monocyte enrichment from PBMCs was achieved via plastic adherence in a 96-well flat bottom CellBIND® microplate (Corning, New York, USA). Supernatant was removed after 3-hours and replaced with either: (i) 100 μL, 0.01% DMSO dissolved in warm RMPI-1640 (Thermo Fisher Scientific, Massachusetts, US) or (ii) 100 μL, 200 μM P2X7 antagonists (vehicle 0.01% DMSO) dissolved in warm RMPI-1640. Enriched monocytes were incubated with pre-treatments for 30-minutes at 37oC in the presence of 5% CO2.

[0148] After this duration, pre-treatments were aspirated, and cells were gently washed with 150 μL warm phosphate-buffered saline (DPBS) without CaCl2 and MgCl2 (Thermo Fisher Scientific, Massachusetts, US). DPBS was aspirated to remove any potential interference from RMPI-1640 or non-adherent populations and replaced with their: (i) 100 μL, 0.01% DMSO dissolved in warm dPBS without CaCl2 and MgCl2, 100 μL, 100 μL, 1 mM BzATP dissolved in 0.01% DMSO in dPBS (HY-136254, MedChemExpress, USA), or 100 μL, 1 mM BzATP in combination with 200 μM P2X7R antagonists in 0.01% DMSO in dPBS.

[0149] Cells were incubated with treatments for 30-minutes at 37oC in the presence of 5% CO2. The supernatant was collected into a clean RNAase and DNAase free Eppendorf and centrifuged at 700 g for 5 -minutes at 4oC.

[0150] Supernatant was then transferred to a clean tube to eliminate any potential contamination from cells carried over from the microplate and samples were stored at -80oC until required for downstream analysis. Human IL-1 beta / IL-1F2 ELISA

[0151] Human IL-1 beta / IL-1F2 were plated in 96-well clear, flat bottom polystyrene microplates with high-bind treatment (R & D Systems, United States) overnight at room temperature. Plates that were not used immediately the next day were stored at 4oC, sealed for up to a week. Before sample preparation, plates were washed with 0.05% Tween® 20 in PBS, pH 7.2-7.4 diluted in Ultrapure water 3-times, removing solution between each wash. Wells were then blocked with 0.1% BSA in PBS, diluted in Ultrapure water for a minimum of 1-hour at room temperature.

[0152] Samples for IL-1β detection were prepared at a 1:4 ratio in 0.1% BSA (same as above blocking buffer) and incubated in the wells for 2-hours at room temperature to achieve adequate capture. After this, samples were removed, and wells were washed with 0.05% Tween® 20 as detailed above. A second antibody aimed to detect the attached protein was incubated for 2-hours at room temperature.

[0153] The plate was washed as detailed above and 1:40 dilution of streptavidin conjugated with horseradish-peroxidase enzyme (Strep-HRP) was diluted in 0.1% BSA and added to the plate for 20-minutes at room temperature. Again, the plate was washed to remove unbound Strep-HRP, and a 1:1 mixture of colour reagent A (known as H2O2) and colour reagent B(Tetramethylbenzidine) was reacting with HRP to induce a colorimetric change from clear to dark blue (colour depends on the amount of cytokine detected) at room temperature. The reaction was stopped after 20-minutes with stop solution (2N H2SO4) and was read on a microplate set to 450 nm, with a wavelength correction set to 540 nm. In vitro Liver Microsomal Stability

[0154] The microsomal stability assay was performed by incubating compounds at 37 °C with liver microsomes sourced from Xenotech (USA). Microsomes were suspended in 0.1 M potassium phosphate buffer (pH 7.4) containing MgCl2 (3.3 mM) . The metabolic reaction was initiated by the addition of NADPH (final concentration 1.3 mM; cofactor required for CYP450 enzyme activity) and quenched at various time points over 60 min by addition of acetonitrile containing an internal standard.

[0155] Control samples with no cofactor were included to assess contribution of NADPH- independent metabolic pathways. Quenched samples were left on ice for approximately 15 min, centrifuged and the supernatant removed and analysed by LC / MS. The in vitro intrinsic clearance (µL / min / mg) was calculated from the first order degradation rate constant (min-1).

[0156] A statistical analysis was performed to confirm that the degradation slope was statistically different to zero. The standard error of the estimated slope was <20% unless stated otherwise. The lower and upper limits for CLint under the current assay conditions are approximately <3 and >920 µL / min / mg protein, respectively. Substrate concentration; 0.5 µM; Protein concentration: 0.5 mg / mL.

[0157] The incubation and data analysis method, together with key assumptions of the general microsomal approach, were based on the following papers: Prediction of human clearance of twenty-nine drugs from hepatic microsomal intrinsic clearance data: An examination of in vitro half-life approach and nonspecific binding to microsomes. Obach RS, Drug Metab Dispos.27:1350–1359, 1999; Measurement of Michaelis constants for cytochrome P450-mediated biotransformation reactions using a substrate depletion approach. Obach RS and Reed-Hagen AE, Drug Metab Dispos, 30:831–837, 2002; Predicting clearance in humans from in vitro data. Obach RS, Curr Topics Med Chem, 11: 334-339, 2011. In vitro Hepatocyte Stability

[0158] The hepatocytes used in this study were sourced from XenoTech (USA) and stored in liquid nitrogen until use. Cryopreserved hepatocytes were suspended in protein-freeincubation buffer (Krebs-Henseleit buffer, pH 7.4, supplemented with calcium chloride (2.5 mM), sodium bicarbonate (25 mM), glycine (3 mM) and fructose (10 mM)) at the target viable cell concentration of 0.5 million cells / mL. The hepatocyte cell viability was assessed using Trypan blue dye exclusion method.

[0159] The metabolic reaction was initiated by addition of compounds at the target substrate concentration (test or QC cocktail) to aliquots of hepatocyte suspension that were pre- equilibrated (for 10 min) at 37°C and 7.5% CO2. At various time points over 240 min, samples were quenched by addition of acetonitrile containing an internal standard. Quenched samples were left on ice for approximately 15 min, centrifuged and the supernatant removed and analysed by tandem quadrupole-Time of Flight MS (Waters G2 QToF) with a mass range scan of 50-1200 Da.

[0160] The in vitro intrinsic clearance (µL / min / million cells) was calculated from the first order degradation rate constant (min-1). A statistical analysis was performed to confirm that the degradation slope was statistically different to zero. The standard error of the estimated slope was <20% unless stated otherwise. The lower and upper limits for CLint under the current assay conditions (0.5 million cells / mL and 240 min incubation period) are approximately <1 and >570 µL / min / million cells, respectively.

[0161] The incubation and data analysis method was based on the following papers: Comparison of Intrinsic Clearance in Liver Microsomes and Hepatocytes from Rats and Humans: Evaluation of Free Fraction and Uptake in Hepatocytes. Lu et al, Drug Metab Dispos, 34:1600–1605, 2006; Simultaneous Assessment In Vitro of Transporter and Metabolic Processes in Hepatic Drug Clearance: Use of a Media Loss Approach. Harrison et al, Drug Metab Dispos, 46:405–414, 2018. Results

[0162] Pharmacological data relevant to this project. In vitro P2X7R antagonism data are described in Table 2, Table 4 and Figure 1. In vitro stability data are described in Table 3, Table 5 and Table 6Table 2. In vitro P2X7R antagonist potency for AZD9056 (R1= R2= R3= H) se, )c5b Human P2X7R IL-1β release assay. THP-1 cells, ELISA kit. c Human P2X7R IL-1β release assay. THP-1 cells, ELISA kit. Table 3. In vitro liver microsomal stability of AZD9056 (R1= R2= R3= H) in vitro CLintTable 4. In vitro P2X7R antagonist potency for the “PKT400” series I I I e, c. , .bHuman P2X7R IL-1β release assay. THP-1 cells, ELISA kit. cHuman P2X7R IL-1β release assay. THP-1 cells, ELISA kit.Table 5. In vitro liver microsomal stability of the “PKT400” series in vitro CLint in)Table 6. In vitro hepatocyte stability of the “PKT400” series in vitro CLintlls)

[0163] The following in vivo models for CVD are proposed to assess AZD9056 and “PKT400” series. Tandem stenosis (duration: approximately 3-months)

[0164] This model aims to evaluate the efficacy of AZD9056 and PKT400 lead(s) in plaque stabilisation after plaque rupture.Description

[0165] In this model, male APOE- / -mice 6-8-weeks old are fed a high fat diet (21% fat, 15% cholesterol) for 6-weeks to promote atherosclerotic plaque development. At 12-weeks of age, mice will undergo a single procedure in which the right carotid artery will be permanently constricted to 150 μm in two distinct regions to induce an unstable phenotype.3The presence of two distinct ligations, the first placed 1 mm from the carotid bifurcation (referred to as distal stenosis) and the second 3 mm proximal to the distal stenosis.4

[0166] The shear stress in the carotid artery is low, while the tensile stress becomes high, particularly upstream from the proximal stenosis,4modelling an important trigger of plaque rupture in humans. At the same time, mice will be allocated to receive either AZD9056, PKT400 lead(s), or vehicle. After this procedure, mice are maintained on the high fat diet for a further 7 weeks. At 20-weeks of age, mice are euthanised and the right carotid artery is removed for downstream histological and molecular analysis. Myocardial infarction (duration: 28-days to evaluate chronic heart failure)

[0167] This model aims to investigate the potential of AZD9056 and PKT400 lead(s) to prevent heart failure after myocardial infarction. Description

[0168] 8-week-old C57BL6 / J male mice will be maintained of a chow diet for the duration of this protocol. Under anesthesia, mice will be ventilated, and the ribs retracted to expose the left ventricle. The left coronary artery will be identified and ligated above the left coronary bifurcation to induce ischemic injury to the left ventricle.5

[0169] Subsequently, mice will be allocated to receive either AZD9056, PKT400 lead(s), or vehicle to evaluate the potential of AZD9056 and PKT400 lead(s) to prevent against heart failure. Efficacy will be established via echocardiograph measures that will be conducted weekly and invasive hemodynamic measures that will be performed before collection. Mice will be sacrificed, and the heart removed for either histological or molecular analysis. Bleomycin pulmonary hypertension model (maximum 21 days)

[0170] This model aims to evaluate the potential of AZD9056 and PKT400 lead(s) to protect against right ventricular failure due to pulmonary hypertension.Description

[0171] 8-12-week-old C57BL6 / J male mice will be maintained on a chow diet for the duration of this protocol. Under anesthesia, mice will be administered a single oropharyngeal aspiration of 2.0 mg / kg of either bleomycin or saline (used as a control).6

[0172] Subsequently, mice will be allocated to receive either AZD9056, PKT400 lead(s), or vehicle to evaluate the potential of AZD9056 and PKT400 lead(s) to prevent against right ventricular heart failure. Efficacy will be established via echocardiograph measures that will be conducted weekly and invasive hemodynamic measures that will be performed before collection. Mice will be sacrificed, and the heart removed for either histological or molecular analysis. Monocrotaline (duration: 28-days to evaluate chronic heart failure)

[0173] This model aims to evaluate the potential of P2X7R antagonism as a treatment for pulmonary hypertension and right ventricular heart failure in rats. Description

[0174] 10-12-week-old male rats will be maintained on a chow diet for the duration of the protocol. Rats will be allocated to receive either 60 mg / kg of monocrotaline or saline as a control via subcutaneous administration.7Rats will be maintained for 4-weeks and will have echocardiography measures performed weekly. Invasive pressure-volume assessment in the right ventricle will be performed before the heart is removed for downstream molecular and histological assessment.

[0175] After assessment of the efficacy of AZD9056 and PKT400 lead(s) in the above models the present inventors will pick a compound to progress into clinical studies. Synthetic Protocols

[0176] The synthesis and characterisation of the compounds according to the invention are described below. Synthesis of PKT401 2 (3s,5s,7s)-3,5,7-trifluoroadamantane-1-carboxamide

[0177] (3s,5s,7s)-3,5,7-trifluoroadamantane-1-carboxylic acid (2.33 g, 9.94 mmol) 1 and 1,1’-carbonyldiimidazole (4.84 g, 29.83 mmol) were dissolved in anhydrous THF (~0.3 mmol of acid / mL) and stirred under a nitrogen atmosphere for 1 h. The solution was cooled in an ice bath, and aq. ammonia (28% w / v, 0.2 mL per mmol of acid) was added. The reaction mixture was allowed to stir for a further 1.5 h, and THF was removed in vacuo. The crude product was dissolved in dichloromethane, washed with aq. sodium hydroxide (1 M, 3 x 100 mL), aq. hydrochloric acid (1 M, 2 x 150 mL), and water (2 x 150 mL). The organic fraction was dried over MgSO4, and dichloromethane was removed by rotary evaporation to yield 2 (1.02 g, 44%) as a white solid.

[0178] mp 178.5–179.5 °C;1H NMR (400 MHz, DMSO-d6): δ 7.27 (1H, br s, -NHa), 7.10 (1H, br s, -NHb), 2.24–1.97 (6H, m, H1), 1.89 (6H, br s, H2) ppm;13C NMR (101 MHz, DMSO-d6): δ 174.6–174.2 (m), 92.8 (dd, J = 188.8, 15.0 Hz), 46.0–44.7 (m), 42.8 (q, J = 10.9 Hz), 41.6–40.8 (m) ppm;19F NMR (376 MHz, DMSO-d6): δ -140.2 (s) ppm; LRMS (–ESI) m / z: 232 ([M–H]–, 100%). All spectroscopic data for this compound matched that previously reported in the literature.13 ((3s,5s,7s)-3,5,7-trifluoroadamantan-1-yl)methanamine hydrochloride

[0179] 2 (1.42 g, was treated with lithium aluminium hydride (924 mg, 24.34 mmol) at 0 °C with stirring, warmed to room temperature over 30 min, and then heated at reflux for 20 h. The reaction mixture was allowed to cool to room temperature, and then chilled water (0.17 mL / mmol) was added dropwise, followed by aq. sodium hydroxide (4 M, 0.17 mL / mmol), and additional water (0.5 mL / mmol). THF was added to mobilise the viscous slurry, and after stirring for 30 min, MgSO4was added directly to the reaction mixture. The product mixture was filtered through a Celite®plug, washing with dichloromethane, and the filtrate was reduced in vacuo to yield the amine. The amine was dissolved in diethyl ether (1 mL / mmol), and then washed with aq. sodium hydroxide (1 M, 1 mL / mmol), water (1 mL / mmol), brine (1 mL / mmol), dried over MgSO4and filtered. The filtrate was cooled in an ice bath, and then HCl in dioxane (4 M, 0.4 mL / mmol) was added dropwise with stirring to form a white precipitate. The precipitate was collected by vacuum filtration and washed with diethyl ether to afford 3 (907 mg, 58%) as a white solid.

[0180] mp 270 °C (sublimation);1H NMR (400 MHz, DMSO-d6): δ 8.29 (3H, br s, - NH3+), 2.81 (2H, d, J = 4.8 Hz, H3), 2.24–2.12 (3H, m), 2.06–1.91 (3H, m), 1.76 (6H, br s) ppm;13C NMR (101 MHz, DMSO-d6): δ 92.4 (dt, J = 188.5, 15.7 Hz), 46.5 (q, J = 2.2 Hz), 45.4–44.8 (m), 41.5–41.0 (m), 35.6 (q, J = 12.0 Hz) ppm;19F NMR (376 MHz, DMSO-d6): δ -140.8 ppm; LRMS (+ESI) m / z: 220 ([M+H]+, 100%). All spectroscopic data for this compound matched that previously reported in the literature.15 2-chloro-5-iodo-N- - 1-mg, (3 mL) was treated with oxalyl chloride (0.500 mL, 5.83 mmol, 5.15 eq.) followed by DMF (1 drop). After stirring for 1 h, solvent was removed under a stream of dry nitrogen gas. A solution of the crude acid chloride in THF (2 mL) was added dropwise to a cooled (0 °C) solution of triethylamine (0.316 mL, 2.27 mmol, 2.00 eq.) and amine 3 in THF (3 mL). The mixture was allowed to warm to rt and was stirred for 18 h, the solvent was reduced in vacuo, and the residue partitioned between ethyl acetate (100 mL) and water (50 mL). The layers were separated, and the organic phase was washed with aq. HCl (1 M, 2 x 30 ml), sat. aq. NaHCO3 (2 x 30 mL), brine (50 mL), and dried over MgSO4, and then reduced in vacuo. The crude residue was purified by flash chromatography (7.5-35% v / v EtOAc in hexanes) to yield 5 (372 mg, 68%) a white crystalline solid.

[0182] 1H NMR (400 MHz, DMSO-d6): δ 8.61 (t, J = 6.3 Hz, 1H), 7.83 – 7.75 (m, 2H), 7.33 – 7.26 (m, 1H), 3.23 (d, J = 6.3 Hz, 2H), 2.15 (dq, J = 9.3, 4.3 Hz, 3H), 2.02 (dq, J = 9.8, 4.7 Hz, 3H), 1.69 (q, J = 3.0 Hz, 6H).13C NMR (101 MHz, DMSO-d6): δ 165.33, 139.21, 138.89, 136.81, 131.49, 129.76, 92.70 (dt, J = 188.4, 15.4 Hz), 92.49, 47.22, 45.98 – 44.78 (m), 42.19 (d, J = 16.5 Hz), 38.03 (q, J = 11.0 Hz);19F NMR (376 MHz, DMSO-d6): δ -140.35 (s) ppm; LRMS (+ESI) m / z: 505.95 ([M+Na]+, 100%). 6 2-chloro-5-(3-oxopropyl)-N-(((3s,5s,7s)-3,5,7-trifluoroadamantan-1-yl)methyl) benzamide

[0183] A mg, , chloride (37 mg, 0.14 mmol, 1.3 eq.), diisopropylethylamine (25 µL), Pd(II)OAc (0.52 mg) and toluene (0.5 mL) were charged into a flask under inert atmosphere, followed by allyl alcohol (20 µL, 2- 3.0 eq.). The resulting orange suspension was heated to 80 °C, whereupon a dark brown biphasic mixture was obtained. The reaction was heated for 24 h, allowed to cool to rt, aq. HCl (1 mL, 1.0 M) was added, and the mixture was stirred at rt for 30 min. The resulting suspension was extracted with toluene (1 mL), and the organic layer was washed with water (2 mL), dried over MgSO4, and was reduced under a stream of nitrogen. The crude residue was purified by preparative thin-layer chromatography (EtOAc:hexane, 1:1). The material obtained was triturated with DCM and hexane to give the title compound as a white precipitate, which was collected. The yield of reaction was variable (4-10%).

[0184] 1H NMR (300 MHz, CDCl3) δ 9.73 (s, 1H), 7.47 (d, J = 2.0 Hz, 1H), 7.31 – 7.12 (m, 2H), 3.41 (d, J = 6.7 Hz, 2H), 2.87 (d, J = 7.0 Hz, 2H), 2.75 (d, J = 7.1 Hz, 2H), 2.17 – 1.78 (m, 7H), 1.66 (s, 8H).13C NMR (75 MHz, CDCl3) δ 200.5, 166.6, 140.1, 134.3, 130.4, 129.0, 128.0, 93.0, 90.6, 48.4, 46.5, 44.7, 43.3, 38.2, 27.1.19F NMR (282 MHz, CDCl3): δ - 143.29 ppm; LRMS (+ESI) m / z: 411.97 ([M-H]+, 100%). 7 2-chloro-5-(3-((3-hydroxypropyl)amino)propyl)-N-(((3s,5s,7s)-3,5,7-trifluoroadamantan- 1-yl)methyl)benzamide hydrochloride

[0185] A solution of 6 (21 mg, 0.05 mmol, 1.0 eq.) in methanol (0.2 mL) was treated with aminopropanol (5.0 µL, 1.1 eq.) and acetic acid (1 drop). The mixture was stirred for 30 min, sodium cyanoborohydride (5.5 µL, 2.0-3.0 eq.) was added, and the mixture was stirred at rt for 2 days. The solvent was subsequently removed in vacuo and the resulting residue was redissolved in CH2Cl2. The organic layer was washed with aq. sat. NaHCO3(2 mL), brine (2 mL), dried over MgSO4 and concentrated under a stream of nitrogen. The crude residue was purified by preparative thin-layer chromatography. The amine collected wasdissolved in Et2O (1 mL) and treated dropwise with HCl (4 M in dioxane, 0.3 mL, 2.0 eq.) with stirring, and the resulting white precipitate was collected, giving the title compound as a HCl salt (2.5 mg, 13%).

[0186] 1H NMR (500 MHz, DMSO-d6) δ 8.88 (s, 1H), 8.59 (t, J = 6.4 Hz, 1H), 7.46 – 7.41 (m, 1H), 7.36 – 7.25 (m, 2H), 4.75 (s, 1H), 3.47 (t, J = 6.0 Hz, 2H), 3.24 (d, J = 6.4 Hz, 2H), 2.96 – 2.89 (m, 2H), 2.89 – 2.83 (m, 2H), 2.69 (t, J = 7.7 Hz, 2H), 2.20 – 1.98 (m, 6H), 1.94 (p, J = 7.9 Hz, 2H), 1.81 – 1.73 (m, 2H), 1.70 (q, J = 3.0 Hz, 6H) ppm;13C NMR (126 MHz, DMSO-d6) δ 166.93, 140.04, 137.05, 130.54, 129.47, 128.68, 127.26, 92.75 (dt, J = 188.3, 15.4 Hz), 57.84, 47.16, 46.17, 45.85 – 45.08 (m), 44.55, 42.30 (dt, J = 15.7, 6.1 Hz), 38.15 (q, J = 10.8 Hz), 31.00, 28.66, 26.92 ppm;19F NMR (376 MHz, Methanol-d4) δ - 143.82 ppm; LRMS (+ESI) m / z: 473.28 ([M+H]+, 100%). HRMS (+ESI) m / z: Calc. for C24H32ClF3N2O2 [M+H]+: 473.21772, found: 473.21764; HPLC 96.73% (254 nm, 17.55 min), 94.33% (230 nm, 17.62 min). Synthesis of PKT402 9 (1r,3R,5S,7r)-3,5-difluoroadamantane-1-carboxamide

[0187] Compound 9 was prepared in an analogous manner to compound 2.

[0188] LRMS +, ; m / z: Calc. for C11H15F2NO [M+Na]+: 238.10139, found: 238.10128.

[0189] NMR spectroscopic data for this compound matched those previously reported in the literature.110 ((1r,3R,5S,7r)-3,5-difluoroadamantan-1-yl)methanamine hydrochloride

[0190] Compound 10 was prepared in an analogous manner to compound 3.

[0191] LRMS +, .

[0192] NMR spectroscopic data for this compound matched that previously reported in the literature.111 2-chloro-N-(((1r,3R,5S,7r)-3,5-difluoroadamantan-1-yl)methyl)-5-iodobenzamide

[0193] Compound 11 was prepared in an analogous manner to compound 5.

[0194] 1H = 1H), 7.78 (dd, J = 8.4, 2.2 Hz, 1H), 7.74 (dd, J = 6.4, 2.2 Hz, 1H), 7.30 (d, J = 8.5 Hz, 1H), 3.14 (d, J = 6.4 Hz, 2H), 2.41 (tp, J = 6.3, 3.2 Hz, 1H), 2.00 (q, J = 5.6 Hz, 2H), 1.82 – 1.56 (m, 8H), 1.37 (d, J = 3.4 Hz, 2H) ppm;13C NMR (126 MHz, DMSO-d6) δ 165.23, 139.14, 139.06, 136.82, 131.51, 129.78, 94.45 (d, J = 13.8 Hz), 92.97 (d, J = 13.7 Hz), 92.49, 48.12, 46.98 (t, J = 18.8 Hz), 43.96 – 43.19 (m), 40.35 (t, J = 9.8 Hz), 36.65, 34.73 (d, J = 7.2 Hz), 30.02 (t, J = 10.5 Hz) ppm;19F NMR (376 MHz, DMSO-d6) δ -133.35 ppm; LRMS (-ESI) m / z: 463.96 ([M-H]-, 100%). 12 2-chloro-N-(((1r,3R,5S,7r)-3,5-difluoroadamantan-1-yl)methyl)-5-(3-oxopropyl) benzamide

[0195] Compound 12 was prepared in an analogous manner to compound 6.

[0196] 1H , = 1H), 7.32 (d, J = 8.2 Hz, 1H), 7.21 (dd, J = 8.2, 2.1 Hz, 1H), 6.36 (s, 1H), 3.38 (d, J = 6.7 Hz, 2H), 3.01 – 2.75 (m, 4H), 2.56 – 2.43 (m, 1H), 2.08 (t, J = 5.3 Hz, 2H), 1.85 – 1.70 (m, 8H), 1.45 (s, 2H) ppm;13C NMR (75 MHz, Chloroform-d) δ 200.70, 166.70, 140.14, 134.82, 131.82, 130.56, 128.23, 94.54 (d, J = 13.6 Hz), 92.05 (d, J = 14.1 Hz), 49.37, 47.68, 44.88, 44.28 (d, J = 17.2 Hz), 40.60, 37.64, 30.56, 27.29 ppm;19F NMR (282 MHz, MeOD) δ -136.7 ppm; LRMS (-ESI) m / z: 393.83 ([M-H]-, 100%). 13 2-chloro-N-(((1r,3R,5S,7r)-3,5-difluoroadamantan-1-yl)methyl)-5-(3-((3- hydroxypropyl)amino)propyl)benzamide hydrochloride

[0197] Compound 13 was prepared in an analogous manner to compound 7.

[0198] , , = 8.2 Hz, 1H), 7.22 (d, J = 2.0 Hz, 1H), 3.59 (t, J = 5.6 Hz, 2H), 3.19 (d, J = 9.8 Hz, 2H), 3.02 (t, J = 7.3 Hz, 4H), 2.96 (m, 4H), 2.66 (t, J = 7.7 Hz, 2H), 2.53 (s, 2H), 2.47 (m, H), 2.05 (t, J = 5.4 Hz, 2H), 1.78 (m, 4H), 1.60 (m, 4H), 1.31 (br s, 2H) ppm;13C NMR (75 MHz, MeOD) δ 169.06, 139.74, 136.60, 130.75, 129.78, 128.43, 128.21, 93.88, 91.40, 58.90, 45.93, 43.77, 40.60, 40.20, 37.22, 31.25, 30.59, 28.21, 27.26 ppm;19F NMR (282 MHz, MeOD) δ 137.24 ppm; LRMS (+ESI) m / z: 455.22 ([M+H]+, 100%); HRMS (+ESI) m / z: Calc. for C24H33ClF2N2O2 [M+H]+: 455.22714, found: 455.22706; HPLC 96.82% (254 nm, 17.67 min), 91.97% (230 nm, 17.66 min). Synthesis of PKT403 15 (1r,3s,5R,7S)-3-fluoroadamantane-1-carboxamide

[0199] Compound 15 was prepared in an analogous manner to compound 2.

[0200] All previously reported in the literature.116 ((1r,3s,5R,7S)-3-fluoroadamantan-1-yl)methanamine hydrochloride

[0201] Compound 16 was prepared in an analogous manner to compound 3.

[0202] All spectroscopic data for this compound matched those previously reported in the literature.117 2-chloro-N-(((1r,3s,5R,7S)-3-fluoroadamantan-1-yl)methyl)-5-iodobenzamide

[0203] Compound 17 was prepared in an analogous manner to compound 5.

[0204] 1H NMR = , 7.77 (dd, J = 8.4, 2.2 Hz, 1H), 7.71 (d, J = 2.2 Hz, 1H), 7.29 (d, J = 8.4 Hz, 1H), 3.03 (d, J = 6.3 Hz, 2H), 2.24 (t, J = 3.9 Hz, 2H), 1.77 (ddt, J = 16.6, 11.6, 5.9 Hz, 5H), 1.64 (d, J = 5.9 Hz, 2H), 1.56 – 1.38 (m, 7H) ppm;13C NMR (126 MHz, DMSO-d6) δ 165.12, 139.22, 139.06, 136.84, 131.52, 129.79, 93.65, 92.45, 92.19, 49.26, 44.91 (d, J = 17.4 Hz), 41.73 (d, J = 17.1 Hz), 38.21 (d, J = 1.8 Hz), 34.64 (d, J = 2.1 Hz), 30.48 (d, J = 9.7 Hz) ppm;19F NMR (471 MHz, DMSO- d6) δ -128.34 ppm; LRMS (-ESI) m / z: 481.98 ([M-Cl]-, 100%); 18 2-chloro-N-(((1r,3s,5R,7S)-3-fluoroadamantan-1-yl)methyl)-5-(3-oxopropyl)benzamide

[0205] Compound 18 was prepared in an analogous manner to compound 6.

[0206] 1H , = Hz, 1H), 7.32 (d, J =8.2 Hz, 1H), 7.22 (d, J =2.1 Hz 1H), 7.19 (d, J =2.1 Hz 1H), 6.32 (s, 1H), 3.29 (d, J =2.1 Hz 2H), 2.95 (t, J = 7.2 Hz, 2H), 2.80 (t, J = 7.3 Hz, 2H), 2.33 (s, 2H), 1.86 (s, 4H), 1.73 (d, J = 5.7 Hz, 2H), 1.61 (d, J = 14.6 Hz, 3H) ppm;13C NMR (75 MHz, CDCl3) δ 1200.61, 166.48, 139.91, 134.97, 133.86, 131.52, 130.41, 128.15, 126.75, 93.94, 91.05, 50.38, 45.35, 41.98, 39.04, 38.81, 35.20, 31.04, 27.19 ppm;19F NMR (282 MHz, MeOD) δ 131.82 ppm; LRMS (-ESI) m / z: 376.16 ([M-H]-, 100%). 19 2-chloro-N-(((1r,3s,5R,7S)-3-fluoroadamantan-1-yl)methyl)-5-(3-((3- hydroxypropyl)amino)propyl)benzamide hydrochloride

[0207] Compound 19 was prepared in an analogous manner to compound 7.

[0208] , = 1H), 7.42 (d, J =8.1 Hz, 1H), 7.33 – 7.23 (m, 2H), 4.74 (s, 1H), 3.47 (t, J = 6.0 Hz, 2H), 3.04 (d, J = 6.3 Hz, 2H), 2.97 – 2.83 (m, 4H), 2.68 (t, J = 7.7 Hz, 2H), 2.25 (p, J = 3.0 Hz, 2H), 1.97 – 1.85 (m, 2H), 1.84 – 1.70 (m, 6H), 1.66 (d, J = 5.9 Hz, 2H), 1.58 – 1.39 (m, 6H) ppm;13C NMR (101 MHz, DMSO-d6) δ 166.66, 139.88, 137.37, 130.32, 129.42, 128.63, 127.27, 93.84, 92.02, 57.85, 49.20, 46.20, 44.96 (d, J = 17.4 Hz), 44.58, 41.75 (d, J = 17.3 Hz), 38.96, 38.27, 34.65, 30.99, 30.49 (d, J = 9.7 Hz), 28.66, 26.93 ppm;19F NMR (376 MHz, DMSO-d6) δ - 128.27 ppm; LRMS (+ESI) m / z: 437.23 ([M+H]+, 100%); HRMS (+ESI) m / z: Calc. for C24H34ClFN2O2 [M+H]+: 437.23656, found: 437.23667; HPLC 97.03% (254 nm, 19.23 min), 97.85% (230 nm, 19.28 min). References (1) Wilkinson, S. M.; Barron, M. L.; O’Brien-Brown, J.; Janssen, B.; Stokes, L.; Werry, E. L.; Chishty, M.; Skarratt, K. K.; Ong, J. A.; Hibbs, D. E.; Vugts, D. J.; Fuller, S.; Windhorst, A. D.; Kassiou, M. Pharmacological Evaluation of Novel Bioisosteres of an Adamantanyl Benzamide P2X 7 Receptor Antagonist. ACS Chem. Neurosci.2017, 8 (11), 2374–2380. https: / / doi.org / 10.1021 / acschemneuro.7b00272. (2) Seeland, S.; Kettiger, H.; Murphy, M.; Treiber, A.; Giller, J.; Kiss, A.; Sube, R.; Krähenbühl, S.; Hafner, M.; Huwyler, J. ATP-Induced Cellular Stress and Mitochondrial Toxicity in Cells Expressing Purinergic P2X7 Receptor. Pharmacol. Res. Perspect.2015, 3 (2), e00123. https: / / doi.org / 10.1002 / prp2.123. (3) Chen, Y.-C.; Rivera, J.; Peter, K. Tandem Stenosis to Induce Atherosclerotic Plaque Instability in the Mouse; Andrés, V., Dorado, B., Eds.; Methods in Molecular Biology; Springer New York: New York, NY, 2015; Vol.1339, pp 333–338. https: / / doi.org / 10.1007 / 978-1-4939-2929-0. (4) Chen, Y.-C.; Bui, A. V.; Diesch, J.; Manasseh, R.; Hausding, C.; Rivera, J.; Haviv, I.; Agrotis, A.; Htun, N. M.; Jowett, J.; Hagemeyer, C. E.; Hannan, R. D.; Bobik, A.; Peter, K. A Novel Mouse Model of Atherosclerotic Plaque Instability for Drug Testing and Mechanistic / Therapeutic Discoveries Using Gene and MicroRNA Expression Profiling. Circ. Res.2013, 113 (3), 252–265. https: / / doi.org / 10.1161 / CIRCRESAHA.113.301562. (5) De Villiers, C.; Riley, P. R. Mouse Models of Myocardial Infarction: Comparing Permanent Ligation and Ischaemia-Reperfusion. Dis. Model. Mech.2020, 13 (11), 1– 5. https: / / doi.org / 10.1242 / dmm.046565. (6) Hansen, T.; Karimi Galougahi, K.; Besnier, M.; Genetzakis, E.; Tsang, M.; Finemore,M.; O’Brien-Brown, J.; Di Bartolo, B. A.; Kassiou, M.; Bubb, K. J.; Figtree, G. A. The Novel P2X7 Receptor Antagonist PKT100 Improves Cardiac Function and Survival in Pulmonary Hypertension by Direct Targeting of the Right Ventricle. Am. J. Physiol. Circ. Physiol.2020, 319 (1), H183–H191. https: / / doi.org / 10.1152 / ajpheart.00580.2019. (7) Bueno-Beti, C.; Sassi, Y.; Hajjar, R. J.; Hadri, L. Pulmonary Artery Hypertension Model in Rats by Monocrotaline Administration. In Methods in Molecular Biology; 2018; Vol.1816, pp 233–241. https: / / doi.org / 10.1007 / 978-1-4939-8597-5_18. Industrial Applicability

[0209] As the P2X7R has been implicated in a wide range of disease states for which no good treatment currently exists (e.g., stroke, ALS, MS, Alzheimer’s disease, Huntington’s disease, atherosclerosis, diabetic retinopathy, dilated cardiomyopathy, ischemic injury and left ventricular hypertrophy post myocardial infarction), development of a small molecule capable of modulating the signaling pathway is of significant commercial interest. In fact, the diverse involvement of the P2X7R in disease ensures any compound developed from this invention has no shortage of options for therapeutic application.

[0210] The inventive series of “PKT400” analogues are novel and have therefore never been explored in any previous P2X7R CVD study. This series has to date revealed valuable pharmacophoric criteria for potent P2X7R inhibition within this scaffold.

[0211] To the best of the Inventors’ knowledge, there are no clinical trials underway investigating inhibitors of the P2X7R for any cardiovascular disease. There is one trial currently underway for Major Depressive Disorder (Inflammation) by Janssen (Phase 2 clinical trial with API JNJ-54175446).

Claims

THE CLAIMS DEFINING THE INVENTION ARE AS FOLLOWS:- 1. A compound of Formula (I): (I)are or with the proviso that R1, R2and R3cannot each be H; or a pharmaceutically-acceptable salt thereof.

2. A compound according to claim 1, wherein R1, R2 and R3 are each F.

3. A compound according to claim 1, wherein R1 and R2 are F and R3 is H.

4. A compound according to claim 1, wherein R1is F and R2and R3are H.

5. A compound according to any one of the preceding claims wherein the pharmaceutically-acceptable salt is a hydrochloride salt.

6. A compound according to any one of the preceding claims, wherein the compound is a P2X7 receptor antagonist.

7. A pharmaceutical composition comprising a compound of Formula (I) according to any one of claims 1 to 6 or a pharmaceutically-acceptable salt thereof, together with a pharmaceutically acceptable carrier, diluent or excipient.

8. A method for the treatment of a disease or condition associated with P2X7R in a subject in need thereof, the method comprising administration to the subject of a therapeutically effective amount of a compound according to any one of claims 1 to 6, a pharmaceutically-acceptable salt thereof, or a composition according to claim 7.

9. A method according to claim 8, wherein the disease or condition is associated with upregulation or increased expression of P2X7R.

10. A method according to claim 8 or claim 9, wherein the disease or condition is associated with activation of P2X7R.

11. A method according to any one of claims 8 to 10, wherein the disease or condition is a neuroinflammatory disease or a neurodegenerative disease.

12. A method according to any one of claims 8 to 11, wherein the disease or condition is a disease of the central nervous system.

13. A method according to any one of claims 8 to 12, wherein the disease or condition is selected from the group consisting of: pain, rheumatoid arthritis, osteoarthritis, sepsis, psoriasis, allergic dermatitis, asthma, chronic obstructive pulmonary disease, airways hyper-responsiveness, septic shock, epilepsy, glomerulonephritis, irritable bowel syndrome, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, coronary artery disease, cardiovascular disease, acute coronary syndrome, myocarditis, pericarditis, atherosclerosis, myocardial ischemia, reperfusion injury, cancer, myeloblastic leukemia, diabetes, Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, Huntington’s disease, glaucoma, multiple sclerosis, amyotrophic lateral sclerosis, depression, age-related macular degeneration, uveitis, neuropathic pain, depression, bipolar affective disorders, anxiety, meningitis, traumatic brain injury, acute spinal cord injury, neuropathic pain, osteoporosis, burn injury, ischemic heart disease, myocardial infarction, stroke, varicose veins, tandem stenosis, bleomycin pulmonary hypertension and monocrotaline pulmonary hypertension.

14. A method according to any one of claims 8 to 13, wherein the disease or condition is selected from the group consisting of: cardiovascular disease, atherosclerosis, myocardial infarction, tandem stenosis, bleomycin pulmonary hypertension and monocrotaline pulmonary hypertension.

15. A method according to any one of claims 8 to 14, wherein the disease or conditionis cardiovascular disease.

16. Use of a compound according to any one of claims 1 to 6, a pharmaceutically- acceptable salt thereof, or a composition according to claim 7, in the manufacture of a medicament for the treatment or prevention of a disease or condition associated with P2X7R in a subject.

17. Use according to claim 16, wherein the disease or condition is associated with upregulation or increased expression of P2X7R.

18. Use according to claim 16 or claim 17, wherein the disease or condition is associated with activation of P2X7R.

19. Use according to any one of claims 16 to 18, wherein the disease or condition is a neuroinflammatory disease or a neurodegenerative disease.

20. Use according to any one of claims 16 to 19, wherein the disease or condition is a disease of the central nervous system.

21. Use according to any one of claims 16 to 20, wherein the disease or condition is selected from the group consisting of: pain, rheumatoid arthritis, osteoarthritis, sepsis, psoriasis, allergic dermatitis, asthma, chronic obstructive pulmonary disease, airways hyper-responsiveness, septic shock, epilepsy, glomerulonephritis, irritable bowel syndrome, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, coronary artery disease, cardiovascular disease, acute coronary syndrome, myocarditis, pericarditis, atherosclerosis, myocardial ischemia, reperfusion injury, cancer, myeloblastic leukemia, diabetes, Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, Huntington’s disease, glaucoma, multiple sclerosis, amyotrophic lateral sclerosis, depression, age-related macular degeneration, uveitis, neuropathic pain, depression, bipolar affective disorders, anxiety, meningitis, traumatic brain injury, acute spinal cord injury, neuropathic pain, osteoporosis, burn injury, ischemic heart disease, myocardial infarction, stroke, varicose veins, tandem stenosis, bleomycin pulmonary hypertension and monocrotaline pulmonary hypertension.

22. Use according to any one of claims 16 to 21, wherein the disease or condition is selected from the group consisting of: cardiovascular disease, atherosclerosis, myocardial infarction, tandem stenosis, bleomycin pulmonary hypertension and monocrotaline pulmonary hypertension.

23. Use according to any one of claims 16 to 22, wherein the disease or condition is cardiovascular disease.

24. A compound according to any one of claims 1 to 6, a pharmaceutically-acceptable salt thereof, or a composition according to claim 7, for use in the treatment or prevention of a disease or condition selected from the group consisting of: pain, rheumatoid arthritis, osteoarthritis, sepsis, psoriasis, allergic dermatitis, asthma, chronic obstructive pulmonary disease, airways hyper-responsiveness, septic shock, epilepsy, glomerulonephritis, irritable bowel syndrome, inflammatory bowel disease, Crohn’s disease, ulcerative colitis, coronary artery disease, cardiovascular disease, acute coronary syndrome, myocarditis, pericarditis, atherosclerosis, myocardial ischemia, reperfusion injury, cancer, myeloblastic leukemia, diabetes, Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, Huntington’s disease, glaucoma, multiple sclerosis, amyotrophic lateral sclerosis, depression, age-related macular degeneration, uveitis, neuropathic pain, depression, bipolar affective disorders, anxiety, meningitis, traumatic brain injury, acute spinal cord injury, neuropathic pain, osteoporosis, burn injury, ischemic heart disease, myocardial infarction, stroke, varicose veins, tandem stenosis, bleomycin pulmonary hypertension and monocrotaline pulmonary hypertension.

25. A method for the synthesis of a compound according to claim 2, the method comprising the steps of: a) forming (3s,5s,7s)-3,5,7-trifluoroadamantane-1-carboxamide from the reaction of (3s,5s,7s)-3,5,7-trifluoroadamantane-1-carboxylic acid and 1,1’- carbonyldiimidazole; b) reducing the (3s,5s,7s)-3,5,7-trifluoroadamantane-1-carboxamide with lithium aluminium hydride to give ((3s,5s,7s)-3,5,7-trifluoroadamantan-1- yl)methanamine hydrochloride;c) reacting the ((3s,5s,7s)-3,5,7-trifluoroadamantan-1-yl)methanamine hydrochloride with 2-chloro-5-iodobenzoic acid, oxalyl chloride, DMF and then triethylamine to give 2-chloro-5-iodo-N-(((3s,5s,7s)-3,5,7-trifluoroadamantan-1- yl)methyl)benzamide; d) reacting the 2-chloro-5-iodo-N-(((3s,5s,7s)-3,5,7- trifluoroadamantan-1-yl)methyl)benzamide with tetrabutylammonium chloride, diisopropylethylamine, Pd(II)OAc and toluene to provide 2-chloro-5-(3- oxopropyl)-N-(((3s,5s,7s)-3,5,7-trifluoroadamantan-1-yl)methyl)benzamide; and e) reacting the 2-chloro-5-(3-oxopropyl)-N-(((3s,5s,7s)-3,5,7- trifluoroadamantan-1-yl)methyl)benzamide with aminopropanol, acetic acid and then sodium cyanoborohydride to give 2-chloro-5-(3-((3- hydroxypropyl)amino)propyl)-N-(((3s,5s,7s)-3,5,7-trifluoroadamantan-1- yl)methyl)benzamide hydrochloride.

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