ZA200605785B - Novel compounds - Google Patents

Description

NOVEL COMPOUNDS

The present invention relates to therapeutically active compounds which are xanthine derivatives, processes for the manufacture of said derivatives, pharmaceutical formulations containing the active compounds and the use of the compounds in therapy, particularly in the treatment of diseases where under-activation of the HMT74A receptor contributes to the disease or where activation of the receptor will be beneficial.

Dyslipidaemia is a general term used to describe individuals with aberrant lipoprotein profiles. Clinically, the main classes of compounds used for the treatment of patients with dyslipidaemia, and therefore at risk of cardiovascular disease are the statins, fibrates, bile- acid binding resins and nicotinic acid. Nicotinic acid (Niacin, a B vitamin) has been used clinically for over 40 years in patients with various forms of dyslipidaemia. The primary mode of action of nicotinic acid is via inhibition of hormone-sensitive triglyceride lipase (HSL), 15 . which results in a lowering of plasma non-esterified fatty acids (NEFA) which in turn alters hepatic fat metabolism to reduce the output of LDL and VLDL (low and very low density fipoprotein). Reduced VLDL levels are thought to lower cholesterol ester transfer protein (CETP) activity to result in increased HDL (high density lipoprotein) levels which may be the cause of the observed cardiovascular benefits. Thus, nicotinic acid produces a very : desirable alteration in lipoprotein profiles; reducing levels of VLDL and LDL whilst increasing

HDL. Nicotinic acid has also been demonstrated to have disease modifying benefits, reducing the progression and increasing the regression of atherosclerotic lesions and reducing the number of cardiovascular events in several trials. :

The observed inhibition of HSL by nicotinic acid treatment is mediated by a decrease in cellular cyclic adenosine monophosphate (CAMP) caused by the G-protein-mediated inhibition of adenylyl cyclase. Recently, the G-protein coupled receptors HM74 and HM74A have been identified as receptors for nicotinic acid (PCT patent application WO02/84298;

Wise et. al. J Biol Chem., 2003, 278 (11), 9869-9874). The DNA sequence of human

HM74A may be found in Genbank; accession number AY148884. Two further papers support this discovery, (Tunaru et. al. Nature Medicine, 2003, 9(3), 352-255 and Soga et. al.

Biochem Biophys Res Commun., 2003, 303 (1) 364-369), however the nomenclature differs slightly. In the Tunaru paper what they term human HM74 is in fact HM74A and in the Soga paper HM74b is identical to HM74A. Cells transfected to express HM74A and/or HM74 gain the ability to elicit G; G-protein mediated responses following exposure to nicotinic acid. In mice lacking the homologue of HM74A (m-PUMA-G) nicotinic acid fails to reduce plasma

NEFA levels.

Certain xanthine derivatives have been synthesised and disclosed in the prior art. For 40 example, EP0389282 discloses xanthine derivatives as potential mediators of cerebrovascular disorders. A range of xanthine derivatives were identified as adenosine receptor antagonists by Jacobson et. al. in J. Med. Chem., 1993, 36, 2639-2644.

We now present a group of xanthine derivatives which are selective agonists of the nicotinic acid receptor HM74A and are thus of penefit in the treatment, prophylaxis and suppression of diseases where under-activation of this receptor either contributes to the disease or where activation of the receptor will be beneficial.

Summary of the Invention

The present invention provides therapeutically active xanthine derivatives and the use of these derivatives in therapy, particularly in the treatment of diseases where under-activation of the HM74A receptor contributes to the disease or where activation of the receptor will be beneficial, in particular diseases of lipid metabolism including dyslipidaemia or hyperlipoproteinaemia such as diabetic dyslipidaemia and mixed dyslipidaemia, heart failure, hypercholesteraemia, cardiovascular disease including atherosclerosis, arteriosclerosis, and hypertriglyceridaemia. As such, the compounds may also find favour as therapeutics for coronary artery disease, thrombosis, angina, chronic renal failure, peripheral vascular disease and stroke, as well as the cardiovascular indications associated with type |i diabetes mellitus, type | diabetes, insulin resistance, hyperlipidaemia, anorexia nervosa, obesity. The compounds may also be of use in the treatment of inflammatory diseases or conditions, as set out further below.

Intermediates, formulations, methods and processes described herein form further aspects of the invention.

Detailed Description of the Invention

According to one aspect of this invention, we provide a compound of Formula (1) 0]

H

R1

SN N

BY | )—rs . 0 N N

R2 0) and a physiologically functional derivative thereof, wherein

R’ is selected from: hydrogen and C., alkyl which may be optionally substituted with one or more groups selected from CN and CFs;

R? is selected from: Ca4o unsubstituted alkyl, C110 alkyl substituted with one or more groups selected from fluorine and CN, Cs alkenyl, unbranched C, alkenyl, and C, alkyl substituted with cycloalkyl; and R? is selected from halogen and CN; with the proviso that: : (i) when R® represents Cl, and R’ represents ethyl, R? is other than propyl; (ii) when R® represents Br, and R' represents propyl, R2? is other than propyl; (ili) when R® represents Cl or Br, and R! represents butyl, R? is other than butyl; and (iv) when R' represents C4 alkyl, CH.CN, or (CH)sCFs3, R2 is other than branched alkyl.

The compounds are of use in the treatment of diseases where under-activation of the

HM74A receptor contributes to the disease or where activation of the receptor will be beneficial, in particular diseases of lipid metabolism including dyslipidaemia or hyperlipoproteinaemia such as diabetic dyslipidaemia and mixed dyslipidaemia, heart failure, hypercholesteraemia, cardiovascular disease including atherosclerosis, arteriosclerosis, and hypertriglyceridaemia. As such, the compounds may also find favour as therapeutics for coronary artery disease, thrombosis, angina, chronic renal failure, peripheral vascular disease and stroke, as well as the cardiovascular indications associated with type Il diabetes mellitus, type | diabetes, insulin resistance, hyperlipidaemia, anorexia nervosa, obesity. As such the compounds of the present invention may find use as agonists or partial agonists of

HMT74A (HM74A modulators).

In particular embodiments, R' is selected from: hydrogen, Ci alkyl, CH.CN and (CH,);CF;

In more particular embodiments R'is selected from: hydrogen and methyl.

In certain embodiments, R? is selected from: Cs.40 unsubstituted alkyl, Csalkyl with one or more CN substitutions, Ci.io alkyl with one or more fluorine substitutions, Cs alkenyl, unbranched C, alkenyl, and C,.4 alkyl substituted with cycloalkyl. Particularly R? is selected from: Ca.1o unsubstituted alkyl; (CH.):.sCN; Cz. alkyl with one or more fluorine substitutions;

Csalkenyl; and Cy alkyl substituted with cycloalkyl. More particularly R? is selected from Cys unsubstituted n-alkyl, for example pentyl; (CH2)1.3CN, for example, (CH,)CN or (CH2):CN; Cs. , alkyl with one or more fluorine substitutions, in particular where the terminal carbon is fully saturated with fluorine, for example (CH,),.sCF3; and Cs alkenyl, in particular, where there is only one double bond, for example where the double bond is located between the fourth and fifth carbons (terminal alkenyl). in particular embodiments, R® represents halogen. More particularly, R3 is selected from: chlorine and bromine. Most particularly, R® represents chlorine. 40 It is to be understood that the present invention includes any combination of particular embodiments and covers all combinations of particular substituents described hereinabove.

Particular compounds of the present invention include: (8-Chloro-2,6-dioxo-1 2,6,7-tetrahydro-3H-purin-3-yl)acetonitrile, 3-Butyl-8-chloro-3,7-dihydro-1H-purine-2,6-dione,

8-Chloro-1-methyl-3-pentyl-3,7-dihydro-1 H-purine-2,6-dione, 8-Chloro-3-(4,4,4-trifluorobutyl)-3,7-dihydro-1 H-purine-2,6-dione, 8-Bromo-1-methyl-3-pentyl-3,7-dihydro-1 H-purine-2,6-dione, 8-Chloro-3-(3,3,3-trifluoropropyl)-3,7-dihydro-1 H-purine-2,6-dione, 8-Chloro-1-propyl-3-(2,2,2-trifluoroethyl)-3,7-dihydro-1 H-purine-2,6-dione,

3-Butyl-8-chloro-1-methyl-3,7-dihydro-1 H-purine-2,6-dione,

(3-B utyl-8-chioro-2,6-dioxo-2,3,6,7-tetrahydro-1 H-purin-1-yl)acetonitrile, 8-Chloro-3-(2-cyclopropylethyl)-3,7-dihydro-1 H-purine-2,6-dione,

8-Chloro-1 ,3-bis(4,4,4-trifluorobutyl)-3,7-dihydro-1 H-purine-2,6-dione, 4-(8-Chloro-1-methyl-2,6-dioxo-1 ,2,6,7-tetrahydro-3H-purin-3-yl)butanenitrile,

156 8-Chloro-1-ethyl-3-(2,2,2-trifluoroethyl)-3,7-dihydro-1 H-purine-2,6-dione, 1-Methyl-2,6-dioxo-3-pentyl-2,3,6,7-tetrahydro-1 H-purine-8-carbonitrile, 8-Chloro-3-propyi-1-methyl-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(3-methylbutyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-pentyl-3,7-dihydro-1H-purine-2,6-dione,

8-Chloro-3-propyl-3,7-dihydro-1H-purine-2,6-dione, 3-Butyl-1-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1 H-purine-8-carbonitrile, 8-Chloro-3-(4-penten-1-yl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-hexyl-3,7-dihydro-1H-purine-2,6-dione, 4-(8-Chloro-2,6-dioxo-1 ,2.6,7-tetrahydro-3H-purin-3-yl)butanenitrile,

8-Chloro-3-hexyl-1-methyl-3,7-dihydro-1 H-purine-2,6-dione, 3-Butyl-8-chloro-1-ethyl-3,7-dihydro-1 H-purine-2,6-dione, [8-Chloro-3-(2-cyclopropylethyl)-2,6-dioxo-2,3,6,7-tetrahyd ro-1H-purin-1-yljacetonitrile, (8-Chloro-2,6-dioxo-3-propyl-2,3,8,7-tetrahydro-1 H-purin-1-yl)acetonitrile, 8-Chloro-1-(4,4 A-trifluorobutyl)-3-(2,2,2-trifluoroethyl)-3,7-dihydro-1 H-purine-2,6-dione,

8-Chloro-3-(2,2,2-trifluoroethyl)-3,7-dihydro-1H-purine-2,6-dione, 2,2'-(8-Chloro-2,6-diox0-6,7-dihydro-1H-purine-1 ,3(2H)-diyt)diacetonitrile, 8-Chloro-1-methyl-3-(4,4,4-trifluorobuty!)-3,7-dihydro-1 H-purine-2,6-dione, 8-Chloro-3-(2-cyclohexylethyl)-3,7-dihydro-1H-purine-2,6-dione, 1,3-Dibutyl-2,6-dioxo-2,3,6,7-tetrahydro-1 H-purine-8-carbonitrile,

1,3-Dibutyl-8-iodo-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(4-methylpentyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(6-methylheptyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-octyl-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-decyl-3,7-dihydro-1H-purine-2,6-dione,

40 8-Chloro-3-(cyclohexylmethyl!)-3,7-dihydro-1H-purine-2,6-dione, (+/-)-8-Chloro-3-(3-methyl pentyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(2-cyclopentylethyl)-3,7-dihydro-1H-purine-2,6-dione,

8-Chloro-3-(cyclopropylmethyl)-3,7-dihydro-1H-purine-2,6-dione, (+/-)-8-Chloro-3-(2-methylbutyl)-3,7-dihydro-1 H-purine-2,6-dione, (+/-)-8-Chloro-3-(2-methylpentyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(cyclobutylmethyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(cyclopentylmethyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(3-cyclopropylpropyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(2-cyclobutylethyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chioro-3-(4-fluorobutyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(3-fluoropropyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(5-fluoropenty!)-3,7-dihydro-1H-purine-2,6-dione, 4-(8-Chloro-1-methyi-2,6-dioxo-1,2,6,7-tetrahydro-3H-purin-3-yl)butanenitrile, 3-(3-Buten-1-yl)-8-chloro-3,7-dihydro-1H-purine-2,6-dione, 6-(8-Chloro-2,6-dioxo-1,2,6,7-tetrahydro-3H-purin-3-yl)-2,2-dimethylhexanenitrile, 8-Chloro-3-(6-fluorohexyl!)-3,7-dihydro-1H-purine-2,6-dione.

Throughout the present specification and the accompanying claims the words "comprise" and “include” and variations such as “comprises”, “comprising”, "includes" and “including” are to be interpreted inclusively. That is, these words are intended to convey the possible inclusion of other elements or integers not specifically recited, where the context allows.

As used herein, the terms “halogen” or “halo” refer to fluorine, chlorine, bromine and iodine.

As used herein, the term “alkyl” (when used as a group or as part of a group) refers to a straight or branched hydrocarbon chain unless specified otherwise, containing the specified number of carbon atoms. For example, Cs-Ciealkyl means a straight or branched hydrocarbon chain containing at least 3 and at most 10 carbon atoms. Examples of alkyl as used herein include, but are not limited to methyl (Me), ethyl (Et), n-propyl! and i-propyl. The term “n-alkyl” refers specifically to an un-branched hydrocarbon chain.

As used herein, the term “cycloalkyl” refers to a hydrocarbon ring containing between 3 and 6 carbon atoms, comprising no heteroatoms or conjugated double bonds. Examples of cycloalkyl as used herein include, but are not limited to cyclopropyl and cyclohexyl.

As used herein, the term “alkenyl” refers to a straight or branched hydrocarbon chain containing the specified number of carbon atoms which contains one or more double bonds.

As used herein, where a group is referred to as being “substituted” with another group or having “one or more substitutions” unless a particular position for such a substitution is specified it is to be understood that a substitution may be present at any position in the 40 group.

As used herein, the term “physiologically functional derivative” refers to any pharmaceutically acceptable derivative of a compound of the present invention, for example an amide thereof, and includes any pharmaceutically acceptable salt of a compound of formula (1), and any pharmaceutically acceptable solvate of a compound of formula (1) which, upon administration to a mammal, such as a human, is capable of providing (directly or indirectly) a compound of formula (I) or an active metabolite or residue thereof. It will be appreciated by those skilled in the art that the compounds of formula (I) may be modified to provide physiologically functional derivatives thereof at any of the functional groups in the compounds, and that the compounds of formula (I) may be so modified at more than one position.

As used herein, the term “pharmaceutically acceptable” used in relation to an ingredient (active ingredient or excipient) which may be included in a pharmaceutical formulation for administration to a patient, refers to that ingredient being acceptable in the sense of being compatible with any other ingredients present in the pharmaceutical formulation and not being deleterious to the recipient thereof.

As used herein, the term “solvate” refers to a complex of variable stoichiometry formed by a solute (in this invention, a compound of formula (I), a salt thereof or a physiologically functional derivative thereof) and a solvent. Such solvents for the purposes of the present invention may not interfere with the biological activity of the solute. The solvent used may be a pharmaceutically acceptable solvent. Examples of suitable pharmaceutically acceptable solvents include water, ethanol and acetic acid. An example of a solvent that may be used is water, in which case the solvate may be referred to as a hydrate of the solute in question.

It will be appreciated that, for pharmaceutical use, the “salt or solvate” referred to above will be a pharmaceutically acceptable salt or solvate. However, other salts or solvates may find use, for example, in the preparation of a compound of formula (I) or in the preparation of a pharmaceutically acceptable salt or solvate thereof.

Pharmaceutically acceptable salts include those described by Berge, Bighley and

Monkhouse, J. Pharm. Sci., 1977, 66, 1-19. Suitable pharmaceutically acceptable salts include alkali metal salts formed from the addition of alkali metal bases such as alkali metal hydroxides. Examples of suitable alkali metal salts are sodium salt or potassium salt. Other suitable pharmaceutically acceptable salts include alkaline earth metal salts such as calcium salt or magnesium salt, ammonium salts; or salts with organic bases such as ethanolamine, triethanolamine, ethylene diamine, triethylmine, choline and meglumine; or salts with amino acids such as arginine, lysine and histidine.

Compounds of formula (I) are of potential therapeutic benefit in the treatment and 40 amelioration of the symptoms of many diseases of lipid metabolism including dyslipidaemia or hyperlipoproteinaemia such as diabetic dyslipidaemia and mixed dyslipidaemia, heart failure, hypercholesteraemia, cardiovascular disease including atherosclerosis,

arteriosclerosis, and hypertriglyceridaemia, type II diabetes mellitus, type | diabetes, insulin resistance, hyperlipidaemia, anorexia nervosa, obesity. As such, the compounds may also find favour as therapeutics for coronary artery disease, thrombosis, angina, chronic renal failure, peripheral vascular disease and stroke.

Furthermore, it is also believed that the HM74 and HM74A receptors are involved in inflammation. Inflammation represents a group of vascular, cellular and neurological responses to trauma. Inflammation can be characterised as the movement of inflammatory cells such as monocytes, neutrophils and granulocytes into the tissues. This is usually associated with reduced endothelial barrier function and oedema into the tissues.

Inflammation with regards to disease typically is referred to as chronic inflammation and can last up to a lifetime. Such chronic inflammation may manifest itself through disease symptoms. The aim of anti-inflammatory therapy is therefore to reduce this chronic inflammation and allow for the physiological process of healing and tissue repair to progress.

Examples of inflammatory diseases or conditions for which the compounds of the present invention may demonstrate utility include those of the joint, particularly arthritis (e.g. rheumatoid arthritis, osteoarthritis, prosthetic joint failure), or the gastrointestinal tract (e.g. ulcerative colitis, Crohn's disease, and other inflammatory bowel and gastrointestinal diseases, gastritis and mucosal inflammation resulting from infection, the enteropathy provoked by non-steroidal anti-inflammatory drugs), of the lung (e.g. adult respiratory distress syndrome, asthma, cystic fibrosis, or chronic obstructive pulmonary disease), of the heart (e.g. myocarditis), of nervous tissue (e.g. multiple sclerosis), of the pancreas, (e.g. inflammation associated with diabetes melitus and complications thereof, of the kidney (e.g. glomerulonephritis), of the skin (e.g. dermatitis, psoriasis, eczema, urticaria, burn injury), of the eye (e.g. glaucoma) as well as of transplanted organs (e.g. rejection) and multi-organ diseases (e.g. systemic lupus erythematosis, sepsis) and inflammatory sequelae of viral or bacterial infections and inflammatory conditions associated with atherosclerosis and following hypoxic or ischaemic insults (with or without reperfusion), for example in the brain or in ischaemic heart disease.

In particular, the compounds of this invention are useful in the treatment and prevention of inflammation, diabetes and cardiovascular diseases or conditions including atherosclerosis, arteriosclerosis, hypertriglyceridemia, and mixed dyslipidaemia.

Nicotinic acid has a significant side effect profile, possibly because it is dosed at high level (gram quantities daily). The most common side effect is an intense cutaneous flushing. In certain embodiments of the present invention the compounds may exhibit reduced side effects compared to nicotinic acid. HM74A has been identified as a high affinity receptor for 40 nicotinic acid whilst HM74 is a lower affinity receptor. The compounds of the present invention may find use as selective HM74A agonists or partial agonists; in which case they will show greater affinity for HM74A than for HM74.

The potential for compounds of formula (I) to activate HM74A may be demonstrated, for example, using the following enzyme and in vitro whole cell assays:

In-vitro testing

For transient transfections, HEK293T cells (HEK293 cells stably expressing the SV40 large

T-antigen) were maintained in DMEM containing 10% foetal calf serum and 2mM glutamine.

Cells were seeded in 90mm culture dishes and grown to 60-80% confiuence (1 8-24h) prior to transfection. Human HM74A (GenBank™ accession number AY148884) was subcloned in to a mammalian expression vector (pcDNA3; Invitrogen) and transfected using

Lipofectamine reagent. For transfection, 9ug of DNA was mixed with 30ul Lipofectamine in 0.6m! of Opti-MEM (Life Technologies Inc.) and was incubated at room temperature for 30min prior to the addition of 1.6ml of Opti-MEM. Cells were exposed to the

Lipofectamine/DNA mixture for 5h and 6ml of 20% (v/v) foetal calf serum in DMEM was then added. Cells were harvested 48h after transfection. Pertussis toxin treatment was carried out by supplementation into media at 50ngmi™* for 16h. All transient transfection studies involved co-transfection of receptor together with the Gy, G protein, Goa.

For generation of stable cell lines the above method was used to transfect CHO-K1 cells seeded in six well dishes grown to 30% confluence. These cells were maintained in DMEM

F-12 HAM media containing 10% foetal calf serum and 2mM glutamine. 48h post- transfection the media was supplemented with 400ug/ml Geneticin (G418, Gibco) for selection of antibiotic resistant cells. Clonal CHO-K1 cell lines stably expressing HM74A were confirmed by [*°S]-GTPyS binding measurements, following the addition of nicotinic acid.

P2 membrane preparation - Plasma membrane-containing P2 particulate fractions were prepared from cell pastes frozen at —80°C after harvest. All procedures were carried out at 4°C. Cell pellets were resuspended in 1 ml of 10mM Tris-HCI! and 0.1mM EDTA, pH 7.5 (buffer A) and by homogenisation for 20s with a Ultra Turrax followed by passage (5 times) through a 25-gauge needle. Cell lysates were centrifuged at 1,000g for 10 min in a microcentrifuge to pellet the nuclei and unbroken cells and P2 particulate fractions were recovered by microcentrifugation at 16,000g for 30min. P2 particulate fractions were resuspended in buffer A and stored at ~80°C until required. [°S]-GTPS binding - assays were performed at room temperature in 384-well format based on methods described previously, (Wieland, T. and Jakobs, K.H. (1994) Methods Enzymol. 40 237, 3-13). Briefly, the dilution of standard or test compounds were prepared and added to a 384-well plate in a volume of 10ul. Membranes (HM74A or HM74) were diluted in assay buffer (20mM HEPES, 100mM NaCl, 10mM MgCl, pH7.4) supplemented with saponin

(60pg/ml), Leadseeker WGA beads (Amersham; 250pg/well) and 10uM GDP, so that the 20u! volume added to each well contains Sug of membranes. [*°S]-GTPyS (1170 Ci/mmol,

Amersham) was diluted (1:1500) in assay buffer and 20p) added to each well. Following the addition of the radioligand, the plates were sealed, pulse spun and incubated for 4hours at room temperature. At the end of the incubation period the plates were read on a Leadseeker machine (VIEWLUX PLUS; Perkin-Elmer) to determine the levels of specific binding.

In-vivo testing HMT74A agonists were tested in male Spague-Dawley rats (200-250g) which had been fasted for at least 12 hours prior to the study. The compounds were dosed intravenously (5mi/kg) or by oral gavage (10ml/kg). Blood samples (0.3m tail vein bleed) were taken pre-dose and at three times post-dose (times ranging from 15 minutes to 8 hours post-dose). Each blood sample was transferred to a heparin tube (Becton Dickinson Microtainer, PST LH) and centrifuged (10,0009 for 5 minutes) to produce a plasma sample. The plasma samples were assayed for levels of non-esterified fatty acids (NEFA) using a commercially available kit (Randox). Inhibition of plasma NEFA levels, relative to pre-dose levels, was used as a surrogate for HM74A agonist activity.

In order to determine whether HM74A compounds exhibited the flushing response associated with nicotinic acid they were dosed to anaesthetised guinea-pigs. Male Dunkin

Hartley guinea pigs (300-800g) were fasted for 12 hours prior to being anaesthetised with a mixture of Ketamine hydrochloride (Vetalar, 40mg/kg i.m.), Xylazine (Rompun, 8mg/kg im.) and sodium pentobarbitone (Sagatal, 30mg/kg i.p.). Following anaesthesia a tracheostomy was performed and the animals were mechanically ventilated with room air (10-12mlL/kg, 60 breaths/min). A jugular vein, and a carotid artery, were cannulated for intravenous administration of test compound and collection of blood respectively. An infra-red temperature probe (Extech Instruments) was placed 3-5mm from the tip of the left ear.

Temperature measurements were recorded every minute from 5 minutes prior to test compound and up to 40 minutes post-administration of test compound. Data was automatically collected on a Psion computer before being transferred for data analysis within an Excel spreadsheet. Prior to, and at frequent time points after, compound administration, blood samples (0.3ml) were taken via the carotid arterial cannula and transferred to

Microtainer (BD) tubes containing lithium heparin. The samples were mixed thoroughly on a blood roller and then stored on ice prior to centrifugation at 1200g for § minutes.

Nicotinic acid (10mg/kg iv.) produced a mean (+ s.e.m.) increase in ear temperature equivalent to 10.42 + 1.44 (area under curve; arbitary units; n=6). By comparison, the compound of Example 30 (10mg/kg i.v.) produced a mean (+ s.e.m.) increase in ear 40 temperature equivalent to 1.52 + 0.39 (area under curve; arbitary units; n=6), a reduction of 85%.

Compounds according to Formula (1) have been synthesised (see synthetic examples below) and tested in one or more of the assays discussed above. All of the exemplified compounds have a pEC50 of 4.9 (+/- 0.3 log unit) or greater and an efficacy of 30% or greater. Some particular compounds are exemplified below.

General purification and analytical methods:

The mass spectra (MS) were recorded on a Fisons VG Platform mass spectrometer using electrospray positive ionisation [(ES+ve to give MH" and M(NH,)" molecular ions] or electrospray negative ionisation [(ES-ve to give (M-H)" molecular ion] modes. 'H NMR spectra were recorded using a Bruker DPX 400MHz spectrometer using tetramethylsilane as the external standard.

Biotage™ chromatography refers to purification carried out using equipment sold by Dyax

Corporation (either the Flash 40i or Flash 150i) and cartridges pre-packed with KPSil.

Mass directed autoprep refers to methods where the material was purified by high performance liquid chromatography on a HPLCABZ+ 5pm column (5cm x 10mm i.d.) with 0.1% HCO.H in water and 95% MeCN, 5% water (0.5% HCO.H) utilising the following gradient elution conditions: 0-1.0 minutes 5%B, 1.0-8.0 minutes 5—30%B, 8.0-8.9 minutes 30%B, 8.9-9.0 minutes 30—95%B, 9.0-9.9 minutes .95%B, 9.9-10 minutes 95—-0%B at a flow rate of 8ml minutes” (System 2). The Gilson 202-fraction collector was triggered by a

VG Platform Mass Spectrometer on detecting the mass of interest.

Preparative h.p.l.c. refers to methods where the material was purified by high performance liquid chromatography on a HPLCABZ+ Sum column (10cm x 21.2mm id.) with 0.1% 'HCO,H in water (A) and MeCN (0.5% HCO.H) (B) utilising the generic gradient elution conditions expressed as “x to y” gradient with a gradient system as follows: 0-1.45minutes x%B, 1.45-20 minutes x—y%B, 20-24 minutes y—95%B, 24-30 minutes 95%B, 32-34 minutes 95—x%B at a flow rate of 8ml minutes” . The Gilson 233 fraction collector was triggered by UV (254nm).

SPE (solid phase extraction) refers to the use of cartridges sold by International Sorbent

Technology Ltd.

Strata Phenyl SPE refers to the use of cartridges sold by Phenomenex. The compound was loaded onto a cartridge previously conditioned with MeCN and equilibrated with 5% MeCN in water. The compound was eluted with 0.1% HCOH in water and MeCN (0.5% HCO.H) in a 40 suitable gradient on a Combiflash Optix 10.

As indicated above, compounds of Formula (I) may find use in human or veterinary medicine, in particular as activators of HM74A, in the management of dyslipidaemia and hyperlipoproteinaemia.

Thus, there is provided as a further aspect of the present invention a compound of formula (1) or a physiologically functional derivative thereof, for use in human or veterinary medicine, particularly in the treatment of disorders of lipid metabolism including dyslipidaemia or hyperlipoproteinaemia such as diabetic dyslipidaemia and mixed dyslipidaemia, heart failure, hypercholesteraemia, cardiovascular disease including atherosclerosis, arteriosclerosis, and hypertriglyceridaemia, type Il diabetes mellitus, type | diabetes, insulin resistance, hyperlipidaemia, anorexia nervosa, obesity. As such, the compounds are also provided for use in the treatment of coronary artery disease, thrombosis, angina, chronic renal failure, peripheral vascular disease and stroke.

There is provided as a further aspect of the present invention a compound of formula (I) or a physiologically functional derivative thereof, for use in the manufacture of a medicament for the treatment of disorders of lipid metabolism including dyslipidaemia or hyperlipoproteinaemia such as diabetic dyslipidaemia and mixed dyslipidaemia, heart failure, hypercholesteraemia, cardiovascular disease including atherosclerosis, arteriosclerosis, and hypertriglyceridaemia, type Il diabetes mellitus, type | diabetes, insulin resistance, hyperlipidaemia, anorexia nervosa, obesity. As such, the compounds are also provided for use in the treatment of coronary artery disease, thrombosis, angina, chronic renal failure, peripheral vascular disease and stroke. it will be appreciated that references herein to treatment extend to prophylaxis, prevention of recurrence and suppression of symptoms as well as the treatment of established conditions.

According to another aspect of the invention, there is provided the use of a compound of formula (11) 0]

H

R1 ~N N

A Rs 07 N N

R2 and physiologically functional derivative thereof, wherein:

R'is selected from: hydrogen and Cy. alkyl which may be optionally substituted with one or more groups selected from CN and CFs;

R? is selected from: Ca.qo unsubstituted alkyl, Cs.10 alkyl substituted with one or more groups selected from fluorine and CN, Cs alkenyl, unbranched Cs alkenyl, and Cy. alkyl substituted with cycloalkyl; and R? is selected from halogen and CN; in the manufacture of a medicament for the treatment of disorders of lipid metabolism including dyslipidaemia or hyperlipoproteinaemia. In particular, the use is provided of a compound of Formula (11) in the manufacture of a medicament for the treatment of diabetic dyslipidaemia or mixed dyslipidaemia, heart failure, hypercholesteraemia, type |l diabetes mellitus, type | diabetes, insulin resistance, hyperlipidaemia, anorexia nervosa, obesity, coronary artery disease, thrombosis, angina, chronic renal failure, stroke and cardiovascular disease including atherosclerosis, arteriosclerosis, and hypertriglyceridaemia.

In one embodiment of the invention, there is provided a compound of formula (If) for use in the treatment of disorders of lipid metabolism including dyslipidaemia or hyperiipoproteinaemia. In particular, the use is provided of a compound of Formula (1) in the manufacture of a medicament for the treatment of diabetic dyslipidaemia or mixed dyslipidaemia, heart failure, hypercholesteraemia, type I diabetes mellitus, type | diabetes, insulin resistance, hyperlipidaemia, anorexia nervosa, obesity, coronary artery disease, thrombosis, angina, chronic renal failure, stroke and cardiovascular disease including atherosclerosis, arteriosclerosis, and hypertriglyceridaemia.

In particular embodiments, R'is selected from: hydrogen, C4 alkyl, CH.CN and (CH3)sCFa.

In more particular embodiments R! is selected from: hydrogen and methyl.

In certain embodiments R? is selected from: Cs.1 unsubstituted alkyl, Cq.10 alkyl substituted with one or more groups selected from fluorine and CN, Cs alkenyl, unbranched C, alkenyl, and Ci alkyl substituted with cycloalkyl. Particularly, R? is selected from: Cj 40 unsubstituted alkyl, Ciealkyl with one or more CN substitutions, Ci.10 alkyl with one or more fluorine substitutions, Cs alkenyl, unbranched C; alkenyl, and C4 alkyl substituted with cycloalkyl.

More particularly R? is selected from: Ca.40 unsubstituted alkyl; (CH2)1sCN; Cas alkyl with one or more fluorine substitutions; Cs alkenyl; and Ci alkyl substituted with cycloalkyl. Most particularly R? is selected from Cus unsubstituted n-alkyl, for example pentyl; (CH2):1.sCN, for example, (CH2)CN or (CH2)sCN; Cay alkyl with one or more fluorine substitutions, in particular where the terminal carbon is fully saturated with fluorine, for example (CH.)2sCF3; and Cs alkenyl, in particular, where there is only one double bond, for example where the double bond is located between the fourth and fifth carbons (terminal alkenyl).

In particular embodiments, R® represents halogen. More particularly, R® is selected from 40 chlorine and bromine. Most particularly, R® represents chlorine.

Particular compounds for use in the treatment of, or in the manufacture of a medicament for the treatment of disorders of lipid metabolism including dislipidaemia or hyperiipoproteinaemia include:

(8-Chloro-2,6-dioxo-1,2,6,7-tetrahydro-3H-purin-3-yl)acetonitrile, 3-Butyl-8-chloro-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-1-methyl-3-penty!-3,7-dinydro-1H-purine-2,6-dione, 8-Chloro-3-(4,4,4-trifluorobutyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Bromo-1-methyl-3-pentyl-3,7-dihydro-1H-purine-2,6-dione,

8-Chloro-3-(3,3,3-trifluoropropyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chioro-1-propyl-3-(2,2,2-triflucroethyl)-3, 7-dihydro-1H-purine-2,6-dione, 3-Butyl-8-chloro-1-methyi-3,7-dihydro-1H-purine-2,6-dione, (3-Butyl-8-chloro-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1 -yl)acetonitrile, 8-Chloro-3-(2-cyclopropylethyl)-3,7-dihydro-1H-purine-2,6-dione,

8-Chloro-1,3-bis(4,4,4-trifluorobutyl)-3,7-dihydro-1H-purine-2,6-dione, 4-(8-Chloro-1-methyl-2,6-dioxo-1,2,6,7-tetrahydro-3H-purin-3-yl)butanenitrile, 8-Chloro-1-ethyl-3-(2,2,2-trifluoroethyi)-3,7-dihydro-1H-purine-2,6-dione, 1-Methyl-2,6-dioxo-3-pentyl-2,3,6,7-tetrahydro-1H-purine-8-carbonitrile, 8-Chloro-3-propyl-1-methyl-3,7-dihydro-1H-purine-2,6-dione,

8-Chloro-3-(3-methylbutyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-pentyl-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-propyl-3,7-dihydro-1H-purine-2,6-dione, 3-Butyl-1-methyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-8-carbonitrile, 8-Chloro-3-(4-penten-1-yl)-3,7-dihydro-1H-purine-2,6-dione,

8-Chloro-3-hexyl-3,7-dihydro-1H-purine-2,6-dione, 4-(8-Chloro-2,6-dioxo-1,2,6,7-tetrahydro-3H-purin-3-yl)butanenitrile, 8-Chloro-3-hexyl-1-methyl-3,7-dihydro-1H-purine-2,6-dione, 3-Butyl-8-chioro-1-ethyl-3,7-dihydro-1H-purine-2,6-dione, [8-Chloro-3-(2-cyclopropylethyl)-2,6-dioxo-2,3,6,7-tetrahydro-1H-purin-1 -ylJacetonitrile,

(8-Chloro-2,6-dioxo-3-propyl-2,3,6,7-tetrahydro-1H-purin-1-yl)acetonitrile, 8-Chloro-1-(4,4,4-trifluorobutyl)-3-(2,2,2-trifluoroethyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(2,2,2-trifluoroethyl)-3,7-dihydro-1H-purine-2,6-dione, 2,2'-(8-Chloro-2,6-dioxo-6,7:dihydro-1H-purine-1,3(2H)-diyt)diacetonitrile, 8-Chloro-1-methyl-3-(4,4,4-triflucrobutyl)-3,7-dihydro-1H-purine-2,6-dione,

8-Chloro-3-(2-cyclohexylethyl)-3,7-dihydro-1H-purine-2,6-dione, 1,3-Dibutyl-2,6-dioxo-2,3,6,7-tetrahydro-1H-purine-8-carbonitrile, 1,3-Dibutyl-8-iodo-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(4-methylpentyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(6-methylheptyl)-3,7-dihydro-1H-purine-2,6-dione,

40 8-Chloro-3-octyl-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-decyl-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(cyclohexylmethyl)-3,7-dihydro-1H-purine-2,6-dione,

(+/-)-8-Chloro-3-(3-methylpentyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(2-cyclopentylethyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(cyclopropyimethyl)-3,7-dihydro-1H-purine-2,6-dione, (+/-)-8-Chloro-3-(2-methylbutyl)-3,7-dihydro-1 H-purine-2,6-dione, (+/-)-8-Chloro-3-(2-methylpentyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(cyclobutylmethyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(cyclopentyimethyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(3-cyclopropylpropyl)-3,7-dihydro-1 H-purine-2,6-dione, 8-Chloro-3-(2-cyclobutylethyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(4-fluorobutyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chloro-3-(3-fluoropropyl)-3,7-dihydro-1H-purine-2,6-dione, 8-Chiloro-3-(5-flucropentyl)-3,7-dihydro-1H-purine-2,6-dione, : 4-(8-Chloro-1-methyl-2,6-dioxo-1,2,6,7-tetrahydro-3H-purin-3-yl)butanenitrile, 3-(3-Buten-1-yl)-8-chloro-3,7-dihydro-1H-purine-2,6-dione, 6-(8-Chloro-2,6-dioxo-1,2,6,7-tetrahydro-3H-purin-3-y1)-2,2-dimethylhexanenitrile, 8-Chloro-3-(6-fluorohexyl)-3,7-dihydro-1H-purine-2,6-dione, 8-chloro-3-ethyl-1-methy!-3,7-dihydro-1H-purine-2,6-dione.

It is to be understood that this aspect of the present invention includes any combination of particular embodiments and covers all combinations of particular substituents described herein above for compounds of Formula (II).

Additionally, the present invention provides the use of a compound of formula (1) or a physiologically functional derivative thereof, in the manufacture of a medicament for the treatment of inflammatory diseases or conditions of the joint, particularly arthritis (e.g. rheumatoid arthritis, osteoarthritis, prosthetic joint failure), or of the gastrointestinal tract (e.g. ulcerative colitis, Crohn's disease, and other inflammatory bowel and gastrointestinal diseases, gastritis and mucosal inflammation resulting from infection, the enteropathy provoked by non-steroidal anti-inflammatory drugs), of the lung (e.g. adult respiratory distress syndrome, asthma, cystic fibrosis, or chronic obstructive pulmonary disease), of the heart (e.g. myocarditis), of nervous tissue (e.g. multiple sclerosis), of the pancreas, (e.g. inflammation associated with diabetes melitus and complications thereof, of the kidney (e.g. glomerulonephritis), of the skin (e.g. dermatitis, psoriasis, eczema, urticaria, bum injury), of the eye (e.g. glaucoma) as well as of transplanted organs (e.g. rejection) and multi-organ diseases (e.g. systemic lupus erythematosis, sepsis) and inflammatory sequelae of viral or bacterial infections and inflammatory conditions associated with atherosclerosis and following hypoxic or ischaemic insults (with or without reperfusion), for example in the brain or in ischaemic heart disease. 10

In a further or altemnative aspect there is provided a method for the treatment of a human or animal subject with a condition where under-activation of the HM74A receptor contributes to the condition or where activation of the receptor will be beneficial, which method comprises administering to said human or animal subject an effective amount of a compound of formula (1) or a physiologically acceptable salt or solvate thereof.

Again, it is to be understood that this aspect of the present invention includes any combination of particular embodiments and covers all combinations of particular substituents described herein above for compounds of Formula (1).

More particularly, the present invention provides a method for the treatment of disorders of lipid metabolism including dyslipidaemia or hyperlipoproteinaemia such as diabetic dyslipidaemia and mixed dyslipidaemia, heart failure, hypercholesteraemia, cardiovascular disease including atherosclerosis, arteriosclerosis, and hypertriglyceridaemia, type II diabetes mellitus, type | diabetes, insulin resistance, hyperlipidaemia, anorexia nervosa, obesity, which method comprises administering to said human or animal subject an effective amount of a compound of formula (1) or a physiologically acceptable salt or solvate thereof.

As such, these compounds may also find favour in methods for the treatment of coronary artery disease, thrombosis, angina, chronic renal failure, peripheral vascular disease and stroke, which methods comprise administering to said human or animal subject an effective amount of a compound of formula (1).

The amount of a HM74A modulator which is required to achieve the desired biological effect will, of course, depend on a number of factors, for example, the mode of administration and the precise clinical condition of the recipient. In general, the daily dose will be in the range of 0.1mg - 1g/kg, typically 0.1 - 100mg/kg. An intravenous dose may, for example, be in the range of 0.01mg to 0.1g/kg, typically 0.01mg to 10mg/kg, which may conveniently be administered as an infusion of from 0.1ug to 1mg, per minute. Infusion fluids suitable for this purpose may contain, for example, from 0.01ug to 0.1mg, per millilitre. Unit doses may contain, for example, from 0.01pg to 1g of a HM74A modulator. Thus ampoules for injection may contain, for example, from 0.01pg to 0.1g and orally administrable unit dose formulations, such as tablets or capsules, may contain, for example, from 0.1mg to 1g. No toxicological effects are indicated/expected when a compound of the invention is administered in the above mentioned dosage range.

A compound of the present invention may be employed as the compound per se in the treatment of a disease where under-activation of the HM74A receptor contributes to the disease or where activation of the receptor will be beneficial, an example of this is where a compound of the present invention is presented with an acceptable carrier in the form of a pharmaceutical formulation. The carrier must, of course, be acceptable in the sense of being compatible with the other ingredients of the formulation and must not be deleterious to 40 the recipient. The carrier may be a solid or a liquid, or both, and may be formulated with the

HM74A modulator as a unit-dose formulation, for example, a tablet, which may contain from 0.05% to 95% by weight of the HM74A modulator.

The formulations include those suitable for oral, rectal, topical, buccal (e.g. sub-lingual) and parenteral (e.g. subcutaneous, intramuscular, intradermal or intravenous) administration.

There is also provided according to the invention a process for preparation of such a pharmaceutical composition which comprises mixing the ingredients.

Formulations suitable for oral administration may be presented in discrete units, such as capsules, cachets, lozenges or tablets, each containing a predetermined amount of a

HMT74A modulator; as a powder or granules; as a solution or a suspension in an aqueous or non-aqueous liquid; or as an oil-in-water or water-in-oil emulsion. In general, the formulations are prepared by uniformly and intimately admixing the active HM74A modulator with a liquid or finely divided solid carrier, or both, and then, if necessary, shaping the product. For example, a tablet may be prepared by compressing or moulding a powder or granules of the HM74A modulator optionally with one or more accessory ingredients.

Compressed tablets may be prepared by compressing, in a suitable machine, the compound in a free-flowing form, such as a powder or granules optionally mixed with a binder, lubricant, inert diluent and/or surface active/dispersing agent(s). Moulded tablets may be made by moulding, in a suitable machine, the powdered compound moistened with an inert liquid diluent.

Tablets and capsules for oral administration may contain conventional excipients such as binding agents, for example syrup, acacia, gelatin, sorbitol, tragacanth, mucilage of starch or polyvinyl pyrrolidone; fillers, for example, lactose, microcrystalline cellulose, sugar, maize- starch, calcium phosphate or sorbitol; lubricants, for example, magnesium stearate, stearic acid, talc, polyethylene glycol or silica; disintegrants, for example, potato starch, croscarmellose sodium or sodium starch glycollate; or wetting agents such as sodium lauryl sulphate. The tablets may be coated according to methods well known in the art. Oral liquid preparations may be in the form of, for example, aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be presented as a dry product for constitution with water or other suitable vehicle before use. Such liquid preparations may contain conventional additives such as suspending agents, for example, sorbitol syrup, methyl cellulose, glucose/sugar syrup, gelatin, hydroxymethyl cellulose, carboxymethyl celiulose, aluminium stearate gel or hydrogenated edible fats; emulsifying agents, for example, lecithin, sorbitan mono-oleate or acacia; non-aqueous vehicles (which may include edible oils), for example almond oil, fractionated coconut oil, oily esters, propylene glycol or ethyl alcohol; or preservatives, for example, methyl or propyl p-hydroxybenzoates or sorbic acid. The preparations may also contain buffer salts, flavouring, colouring and/or sweetening agents (e.g. mannitol) as appropriate. 40

Formulations suitable for buccal (sub-lingual) administration include lozenges comprising a

HM74A modulator in a flavoured base, usually sucrose and acacia or tragacanth, and pastilles comprising the HM74A modulator in an inert base such as gelatin and glycerin or sucrose and acacia.

Formulations of the present invention suitable for parenteral administration conveniently comprise sterile aqueous preparations of an HM74A modulator, the formulation may be isotonic with the blood of the intended recipient. These preparations could be administered intravenously, although administration may also be effected by means of subcutaneous, intramuscular, or intradermal injection. Such preparations may conveniently be prepared by admixing the HM74A modulator with water and rendering the resulting solution sterile and isotonic with the blood. Injectable compositions according to the invention will generally contain from 0.1 to 5% w/w of the HM74A modulator.

Thus, formulations of the present invention suitable for parenteral administration comprising a compound according to the invention may be formulated for parenteral administration by bolus injection or continuous infusion and may be presented in unit dose form, for instance as ampoules, vials, small volume infusions or pre-filled syringes, or in multi-dose containers with an added preservative. The compositions may take such forms as solutions, suspensions, or emulsions in aqueous Or non-aqueous vehicles, and may contain formulatory agents such as anti-oxidants, buffers, antimicrobial agents and/or toxicity adjusting agents. Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g. sterile, pyrogen-free water, before use. The dry solid presentation may be prepared by filling a sterile powder aseptically into individual sterile containers or by filling a sterile solution aseptically into each container and freeze-drying.

Formulations suitable for rectal administration may be presented as unit-dose suppositories.

These may be prepared by admixing a HM74A modulator with one or more conventional solid carriers, for example, cocoa butter or glycerides and then shaping the resulting mixture.

Formulations suitable for topical application to the skin may take the form of an ointment, cream, lotion, paste, gel, spray, aerosol, or oil. Carriers which may be used include vaseline, lanolin, polyethylene glycols, alcohols, and combinations of two or more thereof.

The HM74A modulator is generally present at a concentration of from 0.1 to 15% w/w of the composition, for example, from 0.5 to 2%.

By topical administration as used herein, we include administration by insufflation and inhalation. Examples of various types of preparation for topical administration include ointments, creams, lotions, powders, pessaries, sprays, aerosols, capsules or cartridges for use in an inhaler or insufflator or drops (e.g. eye or nose drops). 40 Ointments and creams may, for example, be formulated with an aqueous or oily base with the addition of suitable thickening and/or gelling agents and/or solvents. Such bases may thus, for example, include water and/or an oil such as liquid paraffin or a vegetable oil such as arachis oil or castor oil or a solvent such as a polyethylene glycol. Thickening agents which may be used include soft paraffin, aluminium stearate, cetostearyl alcohol, polyethylene glycols, microcrystalline wax and beeswax.

Lotions may be formulated with an aqueous or oily base and will in general also contain one or more emulsifying agents, stabilising agents, dispersing agents, suspending agents or thickening agents.

Powders for external application may be formed with the aid of any suitable powder base, for example, talc, lactose or starch. Drops may be formulated with an aqueous or non-aqueous base also comprising one or more dispersing agents, solubilising agents or suspending agents.

Spray compositions may be formulated, for example, as aqueous solutions or suspensions or as aerosols delivered from pressurised packs, with the use of a suitable propeliant, e.g. dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, 1,1,1,2,3,3,3- heptafluoropropane, 1,1,1,2- tetraflucrethane, carbon dioxide or other suitable gas.

Capsules and cartridges for use in an inhaler or insufflator, of for example gelatin, may be formulated containing a powder mix of a compound of the invention and a suitable powder base such as lactose or starch. :

The pharmaceutical compositions according to the invention may also be used in combination with other therapeutic agents, for example in combination with other classes of dyslipidaemic drugs (e.g. statins, fibrates, bile-acid binding resins or nicotinic acid).

The compounds of the instant invention may be used in combination with one or more other therapeutic agents for example in combination with other classes of dyslipidaemic drugs e.g. 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins) or fibrates or bile acid binding resins or nicotinic acid. The invention thus provides, in a further aspect, the use of such a combination in the treatment of diseases where under-activation of the HM74A receptor contributes to the disease or where activation of the receptor will be beneficial and the use of a compound of formula (I) or (Il) or a pharmaceutically acceptable salt, solvate or physiologically functional derivative thereof in the manufacture of a medicament for the combination therapy of disorders of lipid metabolism including dyslipidaemia or hyperlipoproteinaemia such as diabetic dyslipidaemia and mixed dyslipidaemia, heart failure, hypercholesteraemia, cardiovascular disease including atherosclerosis, arteriosclerosis, and hypertriglyceridaemia, type Il diabetes mellitus, type | diabetes, insulin resistance, hyperlipidaemia, anorexia nervosa or obesity. 10

When the compounds of the present invention are used in combination with other therapeutic agents, the compounds may be administered either sequentially or simultaneously by any convenient route.

The combinations referred to above may conveniently be presented for use in the form of a pharmaceutical formulation and thus pharmaceutical formulations comprising a combination as defined above optimally together with a pharmaceutically acceptable carrier or excipient comprise a further aspect of the invention. The individual components of such combinations may be administered either sequentially or simultaneously in separate or combined pharmaceutical formulations.

When combined in the same formulation it will be appreciated that the two components must be stable and compatible with each other and the other components of the formulation and may be formulated for administration. When formulated separately they may be provided in any convenient formulation, conveniently in such a manner as are known for such compounds in the art.

When in combination with a second therapeutic agent active against the same disease, the dose of each component may differ from that when the compound is used alone.

Appropriate doses will be readily appreciated by those skilled in the art.

The invention thus provides, in a further aspect, a combination comprising a compound of formula (1) or (Il) or a physiologically acceptable salt or solvate thereof together with another therapeutically active agent.

The combination referred to above may conveniently be presented for use in the form of a pharmaceutical formulation and thus pharmaceutical formulations comprising a combination as defined above together with a pharmaceutically acceptable carrier thereof represent a further aspect of the invention.

The compounds of the present invention have useful duration of action.

The compounds of the present invention and salts and solvates thereof may be prepared by the methodology described hereinafter, constituting a further aspect of this invention.

Process A:

A process according to the invention for preparing a compound of formula (1) or formula (11) 40 in which R'is H or is the same as R? and R® is Cl, comprises:

: J J

AN

NY N\ N NaNO, fn x Qe ST

HN" TN N DMSO H OS N ACOH, H,0 A N

HOw 0} (0)

HO OH (in) | DoF

R-X, base

Ri N HN 1 Hi $ DMF L | Sa 8 Cl eee 3 t

Ye ' Paes | w) fas

R2 R2

PA(PPh,), marphaline \))

Ix

Ri N of

Ni . | s—cl pac Ty 2 R2 i) Alkylation of guanine with allyl bromide ii) Diazotisation with sodium nitrite followed by hydrolysis to form the xanthine iii) Chlorination iv) Alkylation at N3 and/or dialkylation at N1 and N3

Vv) Palladium catalysed removal of the allyl group

Process B:

A process according to the invention for preparing a compound of formula (}) or formula (11) in which R® is CN comprises steps (i) and (ii) of Process A followed by:

Af ’ =

HINT N R2-X R1

A I + oe xd R1-X Pr 3 5 8/ ———— [) oN N KiCO, a N Na,CO,or K,CO4 oP; | » (if) R2 ) R2 1) LIHMDS v) DMF " 0) . oJ — 1) HO-NH,. HCI RY

Ri. 1 A _ morpholine Ris 3 I =n Pyridine Fo"

PN 7 =" es M h 2)Ac,0 0 N NB

R2 R2 R2 ill) Alkylation at N3 iv) Alkylation at N1

Vv) Formation of aldehyde at C8 by lithiation with LIHMDS and DMF quench vi) Conversion of the aldehyde to the nitrile vii) Palladium catalysed removal of the allyl group

Process C:

A process according to the invention for preparing a compound of formula (1) or formula (1) in which R® is CI or Br comprises steps (i) to (iv) of Process B followed by:

J

1} NCS or NBS Ri H

Ri N DMF x

NY [) xd > Le ii) PA(PPhs),

R2 ee R2

R3 =ClorBr i) Halogenation at C8 using NCS or NBS ii) Palladium catalysed removal of the allyl group

Process D:

A process according to the invention for preparing a compound of formula (1) or formula (li) in which R® is CN comprises steps (i) to (iv) of Process B followed by: a A J J

Ri oY ; LIOH Ri Ar

Nj 1) HMDS Rio N fo) SN jo] 8 — N: ren ft. 1 R ,

PA » 2) 1 Ar =< MeOHH,O Aus | » - 1 0? >N~ N oO 0” °N

Ra a” To” Re R2 ) )

DIPEA

PyBOP (vii) 2M NH,

DMF

Ar omy Aw c

Ri. of R1 Rig N

NT _ morpholine “N N POCI, Ny

SN. a A — -——— 3 | %

A oo TT Tow A NN,

R2 R2 R2

Vv) Formation of ester vi) Hydrolysis of the methyl ester vii) Conversion of the acid to the amide. viii) Conversion of the amide to the nitrile ix) Palladium catalysed removal of the allyl group

Process E:

A process according to the invention for preparing a compound of formula (I) or formula (ll) in which R® is Cl comprises: lo] 0 HN; N Ring N

HN A K,CO,/ DMF J_s KC, DMF Js

AL | rex Rix ©)

Pd(OH)y/

H, 50 Psi (ii)

Ri A Ri H

Nt N 3 | \ Ap 5 ~~; K NGS! 120°C MeCN. _ A NV

R2 (wv) he i) Alkylation at N3 ii) Alkylation at N1 iii) Debenzylation iv) Chlorination at C8

Process F:

A process according to the invention for preparing a compound of formula (1) or formuia (11) in which R' differs from R? and R® is Cl comprises steps (i) to (iv) of Process A followed by: 0c J

Pd(PPh3)4 oY R1-X, base Ri Morpholine er 8) —C — 1 5 —_— | )—0 oA | 7 CRS a (vi) Pi v

Re R2 R2

Vv) Alkylation at N1 vi) Palladium catalysed removal of the allyl group

Process G:

A process according to the invention for preparing a compound of formula (1) or formula (tn in which R? differs from R2 and R® is Cl comprises steps (i) to (v) of Process F (where R? from process F is specifically SEM or MEM) followed by:

J ~~

Yrs trifluoroacetic acid ~ BE 1) R2-X, base ~~ N

Wa —_— = SPPh, Le oy a Ad am A

SEM/MEM (vil) R2 vi) Cleavage of MEM or SEM protecting group group vii) Alkylation of N3 followed by palladium catalysed removal of the allyl group

Process H:

A process according to the invention for preparing a compound of formula (1) or formula (il) in which R® is Cl, Br, | or F comprises steps (i) to (iv) of Process B followed by: fo) A ai ae q

Ri Pd(Ph,), Ng i N NCS or NBS or NIS Ring

Z| a —_— 7, DE ——— ]

PRE PhSiH A N ove IP a 1 W R2 “ R2

Vv) Palladium catalysed removal of allyl group vi) Halogenation at C8 using NCS, NBS or NIS

Process I:

A process according to the invention for preparing a compound of formula (1) or formula (11) in which R' is H or alkyl, R2is alkyl and R® is Cl comprises: 0 0 7 1 NaOH HN NaNO NO a a HN a — A uz, A { EtOH, oP NT SNH, ACOH HCL ZN Sy

R2 CN . 2 (ii) 1 2 0] R2 R2 - reducing (ii) 0) agent f f i NH 7 Te 3 n

IPN N N R1X, base 0 N N Hol OEY, A 0 %, NH,

R2 R2 : vi nos 2 ou a o# N

R2 i) Pyrimidinedione formation il) Nitrosation iii) Reduction using Na,S;0, or a similar reducing agent iv) Xanthine formation v) Alkylation at N1 (optional) vi) Halogenation at C8 using NCS

Where desired or necessary, as a final stage in any of the above synthetic processes, a resultant compound of formula (I) or (Il) can be converted into a physiologically acceptable salt form or vice versa or converting one salt form into another physiologically acceptable salt form.

ABBREVIATIONS

THF Tetrahydrofuran

Ac Acetyl

DCM Dichloromethane

DMEM Dulbecco's Modified Eagle's Medium

HEPES 4-(2-Hydroxyethyl)piperazine-1-ethanesulphonic acid

DMSO Dimethylsulphoxide

NBS N-bromosuccinimide

NCS N-chlorosuccinimide

NIS N-iodosuccinimide

DMF Dimethyiformamide

LIHMDS Lithium hexamethyldisilylamide

DBAD Dibenzylazodicarboxylate

DIPEA Diisopropylethylamine

PyBOP Benzotriazo-1-yloxytripyrrolidinophosphonium hexafluorophosphate

MEM Methoxyethyloxymethyl

SEM 2-(trimethylsilyl)ethoxymethyl

TFA Trifluoroacetic acid

RT room temperature

FAN Heat

The following non-limiting examples illustrate the present invention:

Claims (26)

. PB60727/ZA January 2008 Claims

1. A compound of Formula (I) 0] H RIN N PR Rs 9) N N R2 0) or a pharmaceutically acceptable salt or solvate thereof, wherein R' is selected from: hydrogen and C,., alkyl which may be optionally substituted with one or more groups selected from CN and CF; R? is selected from: Cs. unsubstituted alky!, C.1o alkyl substituted with one or more groups selected from fluorine and CN, Cs alkenyl, unbranched C, alkenyl, and C,.4 alkyl substituted with cycloalkyl; and R® is selected from halogen and CN; with the proviso that: (i) when R® represents Cl, and R' represents ethyl, R? is other than propyl; (ii) when R® represents Br, and R' represents propyl, R? is other than propyl; (iii) when R® represents Cl or Br, and R' represents butyl, R? is other than butyl; and (iv) when R' represents C,4 alkyl, CH,CN, or (CH,);CF3, R? is other than branched alkyl.

2. A compound according to claim 1 wherein R' is selected from: hydrogen, Cy.4 alkyl, CH,CN and (CH,);CF3; R? is selected from: C19 unsubstituted alkyl, (CH;)..sCN, C,.5 alkyl with one or more fluorine substitutions, Cs alkenyl and C,_, alkyl substituted with cycloalkyl; and R? is selected from halogen and CN; with the proviso that: (i) when R’ represents Cl. and R' represents ethyl, R? is other than propyl; (ii) when R* represents Cl or Br and R' represents butyl, R? is other than butyl; and (iii) when R' represents C,.; alkyl, CH.CN, or (CH.),CF,, R? is other than branched alkyl.

3. A compound according to claim | or 2 wherein R'is selected from: hydrogen and methyl. 66 AMENDED SHEET

. PB60727/ZA January 2008

4. A compound according to any one of claims 1 to 3 wherein R? is selected from: Cas unsubstituted n-alkyl, (CH,).3CN, C4 alkyl with one or more fluorine substitutions and Cs alkenyl.

5. A compound according to any one of claims 1 to 4 wherein R® represents halogen.

6. A compound according to any one of claims 1 to 5 wherein R® is selected from: chlorine and bromine.

7. A compound according to any one of claims 1 to 6 wherein R* represents chlorine.

8. A compound of formula (I) as claimed in claim 1 which is 8-chloro-3-pentyl-3,7- dihydro-1H-purine- 2,6-dione or a pharmaceutically acceptable salt or solvate thereof.

9. A compound as claimed in claim 8 which is 8-chloro-3-pentyl-3,7-dihydro-1H-purine- 2,6-dione.

10. A compound according to any one of claims 1 to 9 for use in human or veterinary medicine.

11. A compound according to any one of claims 1 to 9 for use in the treatment of disorders of lipid metabolism.

12. A compound according to any one of claims 1 to 9 for use in the treatment of diabetic dyslipidaemia, mixed dyslipidaemia, heart failure, hypercholesteraemia, atherosclerosis, arteriosclerosis, and hypertriglyceridaemia, type Il diabetes mellitus, type | diabetes, insulin resistance, hyperlipidaemia, anorexia nervosa, obesity, coronary artery disease, thrombosis, angina, chronic renal failure, peripheral vascular disease or stroke.

13. A compound according to any one of claims 1 to 9 for use in the treatment of dyslipidaemia, hyperlipoproteinaemia, hypercholesteraemia or hypertriglyceridaemia.

14. Use of a compound of Formula (ll) 67 AMENDED SHEET

. PB60727/ZA January 2008 0 H R1 nN N PY )—R3 : o™" "nN N R2 (1) or a pharmaceutically acceptable salt or solvate thereof, wherein R'is selected from: hydrogen and C4 alkyl which may be optionally substituted with one or more groups selected from CN and CF; R? is selected from: C,.1o unsubstituted alkyl, Ci.10 alkyl substituted with one or more groups selected from fluorine and CN, Cs alkenyl, unbranched C, alkenyl, and C,.4 alkyl substituted with cycloalkyl; and R® is selected from halogen and CN: in the manufacture of a medicament for the treatment of a human or animal subject having a condition where under-activation of the HM74A receptor contributes to the condition or where activation of the receptor will be beneficial.

15. Use of a compound of formula (Il) as defined in claim 14 in the manufacture of a medicament for the treatment of disorders of lipid metabolism.

16. Use of a compound of formula (Il) as defined in claim 14 in the manufacture of a medicament for treating diabetic dyslipidaemia, mixed dyslipidaemia, heart failure, hypercholesteraemia, atherosclerosis, arteriosclerosis, and hypertriglyceridaemia, type Il diabetes mellitus, type | diabetes, insulin resistance, hyperlipidaemia, anorexia nervosa, obesity, coronary artery disease, thrombosis, angina, chronic renal failure or stroke.

17. Use of a compound of formula (ll) as defined in claim 14 in the manufacture of a medicament for the treatment of dyslipidaemia, hyperlipoproteinaemia, hypercholesteraemia or hypertriglyceridaemia.

18. A pharmaceutical formulation comprising a compound according to any one of claims 1 to 9 and cne or more physiologically acceptable diluents, excipients or carriers.

19 A combination for administration together or separately. sequentially or simultaneously in separate or combined pharmaceutical formulations, said combination comprising a compound according to any one of claims 1 to 9 together bs) 2th another therapeddtically active agent. 68 AMENDED SHEET

20. A pharmaceutical formulation comprising: (i) a compound according to any one of claims 1-9; (ii) one or more active ingredients selected from statins, fibrates, bile-acid binding resins and nicotinic acid: and (iii) one or more physiologically acceptable diluents, excipients or carriers.

21. A method for the preparation of a compound according to any one of claims 1-9 in which R®is halogen, the method comprising: (i) alkylation at N1 or N3, or dialkylation at N1 and N3 of an N7 protected xanthine; (ii) halogenation at C8; and (iii) de-protection; in any order providing de-protection is carried out after alkylation.

22. A compound according to claim 1, substantially as herein described and exemplified.

23 Use of a compound according to claim 14, substantially as herein described and exemplified.

24. A pharmaceutical formulation according to claim 18 or 20, substantially as herein described and exemplified.

25. A combination according to claim 19, substantially as herein described and exemplified.

26. A method for the preparation of a compound according to claim 21, substantially as herein described and exemplified. 69 AMENDED SHEET