WO2010076564A2 - Isochromenones useful in the treatment of inflammation - Google Patents

Abstract

This invention is directed to compounds of formula (I): wherein R¹, R², R^3, R^x, R^y, m and n have meanings given in the description, or pharmaceutically acceptable salts, solvates or prodrugs thereof, which are useful as PDE4, PDE7 or PDE4 and PDE7 inhibitors, and therefore useful e.g. in the treatment and/or prevention of diseases and conditions associated with inflammation.

Description

ISOCHROMENONES USEFUL IN THE TREATMENT OF INFLAMMATION

Field of the invention

The present invention is directed to substituted isochromenone compounds and pharmaceutical compositions thereof which are useful as inhibitors of phosphodiesterase (PDE) 7 and PDE4. The compounds and compositions are therefore of potential utility in treating inflammatory diseases. The invention also relates to the use of such compounds as medicaments and to synthetic routes for their production.

Background of the invention

Inflammation is an essential localized host response to invading microorganisms or tissue injury which involves cells of the immune system. The classic signs of inflammation include redness (erythema), swelling (edema), pain and increased heat production (pyrema) at the site of injury. The inflammatory response allows the body to specifically recognize and eliminate an invading organism and/or repair tissue injury.

Many of the acute changes at the site of inflammation are either directly or indirectly attributable to the massive influx of leukocytes (e.g., neutrophils, eosinophils, lymphocytes, monocytes) which is intrinsic to this response. Leukocytic infiltration and accumulation in tissue results in their activation and subsequent release of inflammatory mediators such as LTB₄, prostaglandins, TNF-α, IL-1β, IL-8, IL-5, IL-6, histamine, proteases and reactive oxygen species for example.

Normal inflammation is a highly regulated process that is tightly controlled at several levels for each of the cell types involved in the response. For example, expression of the pro-inflammatory cytokine TNF-α is controlled at the level of gene expression, translation, post-translational modification and release of the mature form from the cell membrane. Pro-inflammatory responses are normally countered by endogenous antiinflammatory mechanisms such as generation of IL-10 or IL-4. A characteristic of a normal inflammatory response is that it is temporary in nature and is followed by a resolution phase which brings the state of the tissue back to its prior condition. The resolution phase is thought to involve up-regulation of anti-inflammatory mechanisms as well as down-regulation of the pro-inflammatory processes.

Inflammatory disease occurs when an inflammatory response is initiated that is inappropriate and/or does not resolve in the normal manner but rather persists and results in a chronic inflammatory state. Inflammatory disease may be systemic (e.g. lupus) or localized to particular tissues or organs and exerts an enormous personal and economic burden on society. Examples of some of the most common and problematic inflammatory diseases are rheumatoid arthritis, inflammatory bowel disease, psoriasis, asthma, chronic obstructive pulmonary disease, emphysema, colitis and ischemia-reperfusion injury.

The cyclic nucleotides, cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), play a key role in regulating cell function and phosphodieasterases (PDEs) provide the main route for the degradation of cyclic nucleotides. cAMP is now known to control the functional and genomic responses for a variety of cellular functions triggered by a wide array of receptors (Beavo, J. A. and Brunton, LL, Nat. Rev. MoI. Cell Biol., 3, 710-718 (2002)). Local control of cAMP signaling is affected by a complex pattern of localized synthesis, by adenylate cyclase (AC), and by phosphodiesterase (PDE)-mediated enzymatic degradation.

Regulation of cAMP activity is important in many biological processes, including inflammation, depression and cognitive function. Chronic inflammation is a multitude of heterogeneous diseases characterized in part by activation of multiple inflammatory cells, particularly cells of lymphoid lineage (including T lymphocytes) and myeloid lineage (including granulocytes, macrophages, and monocytes). Activation of these inflammatory cells results in production and release of proinflammatory mediators, including cytokines and chemokines, such as tumor necrosis factor (TNF) and interleukin-1 (IL-1). Discovery of a molecule that suppresses or inhibits such cellular activation and proinflammatory mediator release would be useful in the therapeutic treatment of inflammatory diseases. Elevated cAMP levels suppress inflammatory cell activation. Increased cAMP levels associated with PDE4 inhibition has therefore become a valid potential therapeutic approach to control inflammatory responses and disorders (Beavo et al., "Cyclic Nucleotide Phosphodiesterases: Structure, Regulation and Drug Action," Wiley and Sons, Chichester, pp. 3-14 (1990); Torphy et al., Drug News and Perspectives, 6, pp. 203-214 (1993); Giembycz et al., CHn. Exp. Allergy, 22, pp. 337-344 (1992); and Sanz, M.J., Cortijo, J., Morcillo, E.J., Pharmacol Ther. 106(3):269-97 (2005)).

The PDEs are a family of enzymes that catalyze the hydrolysis of 3',5'-cyclic nucleotides to 5' nucleoside monophosphates, including the conversion of cAMP to AMP and cGMP to GMP. PDE enzymes are collectively grouped as a superfamily of eleven different, but homologous, gene-families with a highly conserved catalytic domain (Soderling, S.H. and Beavo, J.A., Curr. Opin. Cell Biol., 12, 174-179 (2000)). At present twenty-one different mammalian PDE genes have been identified. Many of these genes are expressed in multiple isoforms either by differing initiation sequences or splicing patterns. Differentiation of the enzymes can be achieved on the basis of substrate specificity, kinetic properties and sensitivity to regulatory molecules. PDEs in families 5, 6 and 9 specifically catalyze the hydrolysis of cGMP while PDEs 4, 7 and 8 are specific for cAMP. Enzymes belonging to the other PDE families (1, 2, 3, 10 and 11 ) catalyze the hydrolysis of both cAMP and cGMP with differing kinetics. Different PDE isozymes can have specific tissue, cellular and subcellular distributions and more than one type of PDE is usually present in any given cell. The types of PDEs expressed in a cell, together with their relative proportions and subcellular localization, control the cyclic nucleotide phenotype of that cell.

The PDE4 enzyme is responsible for selective, high affinity hydrolytic degradation of the second messenger cAMP, has a low Michaelis constant and is sensitive to inhibition by rolipram. Recently, a number of selective PDE4 inhibitors have been discovered to have beneficial pharmacological effects resulting from PDE4 inhibition as shown in a variety of disease models (Torphy et al., Environ. Health Perspect, 102 Suppl. 10, 79-84, 1994; Duplantier et al., J. Med. Chem., 39 120-125 (1996); Schneider et al., Pharmacol. Biochem. Behav., 50, 211-217 (1995); Banner and Page, Br. J. Pharmacol., 114, 93-98 (1995); Barnette et al., J. Pharmacol. Exp. Ther., 273, 674-679 (1995); Wright et al., "Differential in vivo and in vitro bronchorelaxant activities of CP-80633, a selective phosphodiesterase 4 inhibitor," Can. J. Physiol. Pharmacol., 75, 1001-1008 (1997); Manabe et al., "Anti-inflammatory and bronchodilator properties of KF19514, a phosphodiesterase 4 and 1 inhibitor," Eur. J. Pharmacol., 332, 97-107 (1997); and Ukita et al., "Novel, potent, and selective phosphodiesterase-4 inhibitors as antiasthmatic agents: synthesis and biological activities of a series of 1- pyridylnaphthalene derivatives," J. Med. Chem., 42, 1088-1099 (1999)).

PDE4 inhibitors have recently shown clinical utility in mitigating the effects of the chronic pulmonary inflammatory diseases of asthma and chronic obstructive pulmonary disease (COPD). Roflumilast, a selective PDE4 inhibitor, demonstrated improvements in measures of airway function (forced expiratory volume in 1 second; FEVi, and peak expiratory flow; PEF) in mild asthmatics in a recently published clinical trial of 12 weeks duration (Bateman et al., Ann. Allergy Asthma Immunol., 96(5): 679- 86 (2006)). A separate study with roflumilast also demonstrated improvements in airway hyperresponsiveness (AHR) to direct histamine provocation in a similar group of mild asthmatics in response to allergen challenge (Louw et al., Respiration, Sept. 5 2006). Recently published results of a long term (6 month) study of cilomilast treatment in patients with COPD indicated that treatment with a selective PDE4 inhibitor arrested airway function (FEVi) decline in these patients and positively affected their quality of life as measured by the St. Georges Respiratory Questionnaire (Rennard et al., Chest, 129(1 ) 65-66 (2006)).

The clinical usefulness of PDE4 inhibition has also been demonstrated in disorders of the central nervous system. PDE4 inhibition by rolipram improves cognitive function in rodents and was developed as an antidepressant in humans. cAMP acts as a second messenger for neurotransmitters, and thus mediates their cellular responses. The therapeutic effects of PDE4 inhibitors in cognition and depression likely originate from enhancement of the cAMP-dependent cellular responses.

PDE7 was first discovered as a new cAMP-specific PDE from a human glioblastoma cDNA library through complementation of PDE-deficient yeast (S. cerevisiae) (see Michaeli, T. et al., "Isolation and characterization of a previously undetected human cAMP phosphodiesterase by complementation of cAMP phosphodiesterase-deficient saccharomyces cerevisiae," J. Biol. Chem. (1993), Vol. 268, No. 17, pp. 12925-12932). It was first detected in human cells as "rolipram-resistant" cAMP PDE activity distinct from PDE4 in human T-cell lines {i.e., Jurkat and Hut-78) (see lchimura, M. et al., "A new cyclic nucleotide phosphodiesterase isozyme expressed in the T-lymphocyte cell lines", BBRC (1993), Vol. 193, No. 3, pp. 985-990). Two PDE7 genes (7A and 7B) are expressed in mammalian tissue (human, rat, mouse etc.). There have been multiple splice variants identified for these genes (i.e., 7A1 , 7A2 and 7A3 for PDE7A) with different tissue distribution and/or intracellular localization. PDE7A and PDE7B are widely expressed at the mRNA level in mammalian tissues but PDE7A is expressed in cells and tissues relevant to the inflammatory response (i.e., T-cells). PDE7A protein expression in primary cells/tissues is much more restricted than its RNA distribution indicating a high-level of translational/post-translational regulation. PDE7A protein expression has been identified (Western analysis or PDE assay) in human T-cells, B- cells, airway epithelium, neutrophils and monocytes/macrophages (see Li, L. et a/., "CD3-and cd28-dependent induction of PDE7 required for T cell activation", Science (1999), Vol. 283, pp. 848-851 ; Lee, R. et a/., "PDE7A is expressed in human B- lymphocytes and is up-regulated by elevation of intracellular cAMP", Cell. Signal. (2002), Vol. 14, pp. 277-284; Fuhrmann, M. et al., "Identification and function of cyclic nucleotide phosphodiesterase isoenzymes in airway epithelial cells", Am. J. Respir. Cell MoI. Biol. (1999), Vol. 20, pp. 292-302).

PDE7A is expressed as a 52-57 kDa protein in human cells. The catalytic domain of PDE7A has 65-70% amino-acid identity to PDE7B while much lower sequence homology (~30 %) to the next most related cAMP pde, PDE4. PKA phosphorylation sites as well as potential negative regulatory domains have been identified in the 5'- (amino terminus) of the protein. This protein has a high affinity for cAMP (1^=100-200 nM) and is insensitive to known selective inhibitors of other PDE classes. The nonselective PDE inhibitor IBMX inhibits the PDE7A protein with an IC₅₀ of 5-10 μM.

One compelling publication reporting a role for PDE7A in T-cell activation comes from a February 1999 Science publication .wherein it was reported that PDE7A is up- regulated (RNA/protein) by anti-CD3/anti-CD28 antibody co-stimulation in peripheral blood CD4+ T-cells and that this correlates with cAMP depression, and induction of IL- 2 and proliferation (Li, vide supra), wherein it was also demonstrated that proliferation of anti-CD3/CD28-stimulated CD4+ peripheral blood T-cells is inhibited by specific anti- sense oligonucleotide (but not by scrambled oligo) to PDE7A. The conclusion from these findings was that PDE7A was necessary for CD3/CD28-mediated activation of human CD4+ T-cells and would be an attractive therapeutic target for selective modulation of T cell responsiveness. Other groups have reported that non-selective PDE7 inhibitors block anti-CD3/anti- CD28-induced proliferation (see Barnes, M. et al., "The effect of non-specific pde7 inhibitors on primary T-cell proliferation (Abstract), William Harvey Research Conference, December 5-7, 2001 , Porto, Portugal). It was also shown that anti-CD3 or antigen-induced cytokine production, proliferation and IL-2 receptor expression in PBMC from house dust mite sensitive donors was sensitive to PDE7 inhibitors but not PDE4 inhibitors (see Kaminuma, O. et al., "Potential role of phosphodiesterase 7 in human T cell function: comparative effects of two phosphodiesterase inhibitors", Japanese Journal of Pharmacology (2001), Vol. 85 (Supplement 1 ), No. 126 (abstract P-266)).

The inhibition of PDE7 potentiates the ability of a PDE4 inhibitor to suppress LPS- induced cytokine production in monocytes/macrophages, which is described in inter alia Vergne, F. et al., "PDE7 inhibitors: chemistry and potential therapeutic utilities", Annual Reports in Medicinal Chemistry (2005), VoI 40, pp. 227-241.

Compounds that suppress or inhibit PDE7 would be of potential use in the therapeutic treatment of inflammatory diseases.

The listing or discussion of an apparently prior-published document in this specification should not necessarily be taken as an acknowledgement that the document is part of the state of the art or is common general knowledge.

US patent US 6,486,155 disclose various isoquinolinones that may be useful for inhibiting neoplasis and therefore may be of potential use in the treatment of cancerous lesions. Journal article Journal of Medicinal Chemistry (2001), 44(13), 2204-2218 by Ukita et al, Japanese patents JP 2000281654, JP 2000072751 and JP 2000072675, and international patent application WO 98/38168 all disclose isoquinolinone derivatives for use as medicaments, for example as PDE inhibitors. Such isoquinolinones may be prepared via isochromenone intermediates that are substituted in the 3-position with a carboxy group. However, there is no disclosure in any of these documents of isochromenones that are substituted with a substituent other than a carboxy group, or that isochromenones themselves may be useful as PDE inhibitors, and therefore of use in the treatment of inflammation. Journal article Organic Letters (2002), 4 (14), 2317-2320 by Nakamura et al discloses methods for preparing inter alia isochromenones. However, this document does not mention that the compounds disclosed therein may have a medical use ascribed to them, and further, it only discloses 3-carboxy-substituted isochromenones.

Journal article Chemical and Pharmaceutical Bulletin (1991), 39(4), 884-8 by Nonaka et al discloses various tannins that have been isolated from a natural product. However, there is no disclosure in this document of isochromenones.

Journal article Justus Liebigs Annalen der Chemie (1974), (7), 1126-31 discloses various method of preparing isochromenones. However, there is no medical use ascribed to such isochromenones and, further, there only appear to be methods for preparing 3-carboxy derivatives thereof.

Japanese patent documents JP 45023546 and JP 45015263 both disclose various compounds, including isochromenones, for use as antiallergic drugs. However, such isochromenones are necessarily substituted with a carboxy group at the 3-position and, further, there is no mention in either of these documents that isochromenones may be useful as PDE inhibitors.

Summary of the invention

There is provided a compound of formula (I):

wherein: n represents 0, 1 , 2, 3 or 4; m represents 0, 1 , 2 or 3;

R¹ represents hydrogen, -C(OR^4a)(R^4b)₂, -C(R^4b)₃ or -Si(R^6p)₃;

R² and R³ independently represent hydrogen, C_1-I2 alkyl (optionally substituted by one or more substituents selected from A¹), aryl (optionally substituted by one or more substituents selected from A²), heteroaryl (optionally substituted by one or more substituents selected from A³), heterocycloalkyl (optionally substituted by one or more substituents selected from A⁴), -C(O)OR^5a, -C(O)N(R^5b)R^5c, -S(O)₂R^5d or -C(O)R^5e;

each R^4a and R^4b independently represent, on each occasion when used herein, hydrogen or C_1-12 alkyl optionally substituted by one or more substituents selected from A⁵; or any two R^4b substituents when attached to a common carbon atom may be linked together to form a 3- to 12-membered ring, optionally containing one to three heteroatoms, one to three unsaturations (e.g. double or triple bonds) and which ring is optionally substituted with one or more substituents selected from A⁶;

each R^x and R^y independently represents, on each occasion when used herein, A⁷ and/or C_1-12 alkyl optionally substituted by one or more substituents selected from A⁸;

A¹, A², A³, A⁴, A⁵, A⁶, A⁷ and A⁸ independently represent aryl (optionally substituted by one or more substituents selected from B¹), heteroaryl (optionally substituted by one or more substituents selected from B²) heterocycloalkyl (optionally substituted by one or more substituents selected from B³), halo, -CN, -NO₂, -C(O)OR^6a, -C(O)N(R^7a)R^8a, -O-R^6b, -OC(O)R⁶⁰, -N(R^7b)R^8b, -C(O)R^6d, -OS(O)₂R⁶⁶, -OC(O)N(R^7c)R^8c, -N(R^6f)C(O)OR^6g, -N(R^6h)C(O)R⁶ⁱ, -N(R^6j)S(O)_tR^6k, -S(O),OR^6m, -S(O)_pR⁶ⁿ, -S(O)_tN(R^7d)R^8d, -Si(R^6p)₃, -N(R^6q)C(O)N(R^7e)R^8e and/or -OC(O)OR^6r; and/or A¹, A⁴, A⁵, A⁶ and A⁸ may alternatively and independently represent =0;

t represents, on each occasion when used herein, 1 or 2;

p represents, on each occasion when used herein, O₁ 1 or 2; R 5b D^8a

R^8b, R^8c, R^8d and R^8e independently represent, on each occasion when used herein, hydrogen, C₁-₁₂ alkyl (optionally substituted by one or more substituents selected from B⁴), aryl (optionally substituted by one or more substituents selected from B⁵), heteroaryl (optionally substituted by one or more substituents selected from B⁶) or heterocycloalkyl (optionally substituted by one or more substituents selected from B⁷); or any pair of R^5b and R^5c, R^7a and R^8a, R^7b and R^8b, R^7c and R^8c, R^7d and R^8d and R^7e and R^8e may be linked together to form, together with the nitrogen atom to which they are necessarily attached, a 3- to 12-membered ring, optionally containing a further one or two heteroatoms, one or two unsaturations (e.g. double or triple bonds), and which ring is optionally substituted by one or more susbtituents selected from B⁸;

R^5a, R^5d, R^6c, R^6e, R^6g, R^6k, R^6p and R^6r independently represent, on each occasion when used herein, C_M2 alkyl (optionally substituted by one or more substituents selected from B⁴), aryl (optionally substituted by one or more substituents selected from B⁵), heteroaryl (optionally substituted by one or more substituents selected from B⁶) or heterocycloalkyl (optionally substituted by one or more substituents selected from B⁷);

B¹, B², B³, B⁴, B⁵, B⁶, B⁷ and B⁸ independently represent halo, C_1-6 alkyl (optionally substituted by one or more halo atoms), -OR^9a, -N(R^9b)R^10b, -NO₂, -CN, aryl (optionally substituted by one or more substituents selected from E¹), heteroaryl (optionally substituted by one or more substituents selected from E²) and/or heterocycloalkyl (optionally substituted by one or more substituents selected from E³); and/or B³, B⁴, B⁷ and B⁸ may alternatively and independently represent =O;

R^9a, R^9b and R^1Ob independently represent hydrogen or Ci_-6 alkyl optionally substituted by one or substituents selected from E⁴;

E¹, E², E³ and E⁴ independently represent halo, -CN, -OH, -OCi_-6 alkyl (optionally substituted by one or more fluoro atoms) and/or Ci_-6 alkyl (optionally substituted by one or more halo atoms),

or a pharmaceutically acceptable salt, solvate, prodrug or polymorph thereof, which compounds are hereinafter referred to as 'the compounds of the invention'.

Pharmaceutically-acceptable salts include acid addition salts and base addition salts. Such salts may be formed by conventional means, for example by reaction of a free acid or a free base form of a compound of formula I with one or more equivalents of an appropriate acid or base, optionally in a solvent, or in a medium in which the salt is insoluble, followed by removal of said solvent, or said medium, using standard techniques (e.g. in vacuo, by freeze-drying or by filtration). Salts may also be prepared by exchanging a counter-ion of a compound of the invention in the form of a salt with another counter-ion, for example using a suitable ion exchange resin.

Compounds of the invention may contain double bonds and may thus exist as E (entgegen) and Z (zusammen) geometric isomers about each individual double bond. All such isomers and mixtures thereof are included within the scope of the invention.

Compounds of the invention may also exhibit tautomerism. All tautomeric forms and mixtures thereof are included within the scope of the invention.

Compounds of the invention may also contain one or more asymmetric carbon atoms and may therefore exhibit optical and/or diastereoisomerism. Diastereoisomers may be separated using conventional techniques, e.g. chromatography or fractional crystallisation. The various stereoisomers may be isolated by separation of a racemic or other mixture of the compounds using conventional, e.g. fractional crystallisation or chromatographic (e.g. HPLC), techniques. Alternatively the desired optical isomers may be made by reaction of the appropriate optically active starting materials under conditions which will not cause racemisation or epimerisation (i.e. a 'chiral pool' method), by reaction of the appropriate starting material with a 'chiral auxiliary' which can subsequently be removed at a suitable stage, by derivatisation (i.e. a resolution, including a dynamic resolution), for example with a homochiral acid followed by separation of the diastereomeric derivatives by conventional means such as chromatography, or by reaction with an appropriate chiral reagent or chiral catalyst all under conditions known to the skilled person. All stereoisomers and mixtures thereof are included within the scope of the invention. Unless otherwise specified, C_1-q alkyl groups (where q is the upper limit of the range) defined herein may be straight-chain or, when there is a sufficient number (i.e. a minimum of two or three, as appropriate) of carbon atoms, be branched-chain, and/or cyclic (so forming a C_3-q-cycloalkyl group). Such cycloalkyl groups may be monocyclic or bicyclic and may further be bridged. Further, when there is a sufficient number (i.e. a minimum of four) of carbon atoms, such groups may also be part cyclic. Such alkyl groups may also be saturated or, when there is a sufficient number (i.e. a minimum of two) of carbon atoms, be unsaturated (forming, for example, a C₂.q alkenyl or a C_2-q alkynyl group).

The term "halo", when used herein, includes fluoro, chloro, bromo and iodo.

Heterocycloalkyl groups that may be mentioned include non-aromatic monocyclic and bicyclic heterocycloalkyl groups (which groups may further be bridged) in which at least one (e.g. one to four) of the atoms in the ring system is other than carbon (i.e. a heteroatom), and in which the total number of atoms in the ring system is between three and twelve (e.g. between five and ten). Further, such heterocycloalkyl groups may be saturated or unsaturated containing one or more double and/or triple bonds, forming for example a C₂.q heterocycloalkenyl (where q is the upper limit of the range) or a C_7-q heterocycloalkynyl group. C_2-q heterocycloalkyl groups that may be mentioned include 7-azabicyclo-[2.2.1]heptanyl, 6-azabicyclo[3.1.1]heptanyl, 6- azabicyclo[3.2.1]-octanyl, 8-azabicyclo[3.2.1]octanyl, aziridinyl, azetidinyl, dihydropyranyl, dihydropyridyl, dihydropyrrolyl (including 2,5-dihydropyrrolyl), dioxolanyl (including 1 ,3-dioxolanyl), dioxanyl (including 1,3-dioxanyl and 1 ,4-dioxanyl), dithianyl (including 1 ,4-dithianyl), dithiolanyl (including 1 ,3-dithiolanyl), imidazolidinyl, imidazolinyl, morpholinyl, 7-oxabicyclo[2.2.1]heptanyl, 6-oxabicyclo[3.2.1]-octanyl, oxetanyl, oxiranyl, piperazinyl, piperidinyl, pyranyl, pyrazolidinyl, pyrrolidinonyl, pyrrolidinyl, pyrrolinyl, quinuclidinyl, sulfolanyl, 3-sulfolenyl, tetrahydropyranyl, tetrahydrofuranyl, tetrahydropyridyl (such as 1,2,3,4-tetrahydropyridyl and 1 ,2,3,6- tetrahydropyridyl), thietanyl, thiiranyl, thiolanyl, thiomorpholinyl, trithianyl (including 1 ,3,5-trithianyl) and the like. Substituents on heterocycloalkyl groups may, where appropriate, be located on any atom in the ring system including a heteroatom. Further, in the case where the substituent is another cyclic compound, then the cyclic compound may be attached through a single atom on the heterocycloalkyl group, forming a so-called "spiro"-compound. The point of attachment of heterocycloalkyl groups may be via any atom in the ring system including (where appropriate) a heteroatom (such as a nitrogen atom), or an atom on any fused carbocyclic ring that may be present as part of the ring system. Heterocycloalkyl groups may also be in the N- or S- oxidised form.

For the avoidance of doubt, the term "bicyclic" (e.g. when employed in the context of heterocycloalkyl groups) refers to groups in which the second ring of a two-ring system is formed between two adjacent atoms of the first ring. The term "bridged" (e.g. when employed in the context of heterocycloalkyl groups) refers to monocyclic or bicyclic groups in which two non-adjacent atoms are linked by either an alkylene or heteroalkylene chain (as appropriate).

Aryl groups that may be mentioned include C_6-14 (such as C_6-13 (e.g. C_6-10)) aryl groups. Such groups may be monocyclic or bicyclic and have between 6 and 14 ring carbon atoms, in which at least one ring is aromatic. C_6-14 aryl groups include phenyl, naphthyl and the like, such as 1 ,2,3,4-tetrahydronaphthyl, indanyl, indenyl and fluorenyl. The point of attachment of aryl groups may be via any atom of the ring system. However, when aryl groups are bicyclic or tricyclic, they are linked to the rest of the molecule via an aromatic ring.

Heteroaryl groups that may be mentioned include those which have between 5 and 14 (e.g. 10) members. Such groups may be monocyclic, bicyclic or tricyclic, provided that at least one of the rings is aromatic and wherein at least one (e.g. one to four) of the atoms in the ring system is other than carbon (i.e. a heteroatom). Heterocyclic groups that may be mentioned include acridinyl, benzimidazolyl, benzodioxanyl, benzodioxepinyl, benzodioxolyl (including 1 ,3-benzodioxolyl), benzofuranyl, benzofurazanyl, benzothiazolyl, benzothiadiazolyl (including 2,1 ,3-benzothiadiazolyl), benzoxadiazolyl (including 2,1 ,3-benzoxadiazolyl), benzoxazinyl (including 3,4-dihydro- 2H-1 ,4-benzoxazinyl), benzoxazolyl, benzomorpholinyl, benzoselenadiazolyl (including 2,1 ,3-benzoselenadiazolyl), benzothienyl, carbazolyl, chromanyl, cinnolinyl, furanyl, imidazolyl, imidazo[1 ,2-a]pyridyl, indazolyl, indolinyl, indolyl, isobenzofuranyl, isochromanyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiaziolyl, isothiochromanyl, isoxazolyl, naphthyridinyl (including 1,6-naphthyridinyl or, preferably, 1,5-naphthyridinyl and 1 ,8-naphthyridinyl), oxadiazolyl (including 1 ,3,4-oxadiazolyl), oxazolyl, phenazinyl, phenothiazinyl, phthalazinyl, pteridinyl, purinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quinolizinyl, quinoxalinyl, tetrahydroisoquinolinyl (including 1,2,3,4-tetrahydroisoquinolinyl and 5,6,7,8- tetrahydroisoquinolinyl), tetrahydroquinolinyl (including 1 ,2,3,4-tetrahydroquinolinyl and 5,6,7,8-tetrahydroquinolinyl), tetrazolyl, thiadiazolyl (including 1 ,3,4-thiadiazolyl), thiazolyl, thiochromanyl, thienyl, triazolyl (including 1 ,2,3-triazolyl and 1 ,2,4-triazolyl) and the like. Substituents on heteroaryl groups may, where appropriate, be located on any atom in the ring system including a heteroatom. The point of attachment of heteroaryl groups may be via any atom in the ring system including (where appropriate) a heteroatom (such as a nitrogen atom), or an atom on any fused carbocyclic ring that may be present as part of the ring system. Heteroaryl groups may also be in the N- or S- oxidised form.

Heteroatoms that may be mentioned include phosphorus, silicon, boron, tellurium, selenium and, preferably, oxygen, nitrogen and sulfur.

For the avoidance of doubt, in cases in which the identity of two or more substituents in a compound of the invention may be the same, the actual identities of the respective substituents are not in any way interdependent. For example, in the situation in which R² and R³ both represent a C_M2 alkyl group optionally substituted as hereinbefore defined, the alkyl groups in question may be the same or different. Similarly, when groups are substituted by more than one substituent as defined herein, the identities of those individual substituents are not to be regarded as being interdependent. For example, when the above-mentioned C^₂ alkyl group is substituted with two A¹ substituents, one of which is -C(O)OR^6a and another which is -O-R^6b, and both R^6a and R^6b represent C_{M ∑} alkyl, then the identities of the two alkyl groups are not to be regarded as being interdependent.

For the avoidance of doubt, -(R^x)_n and -(R^y)_m respectively represent between one and four, and between one and three optional (i.e. R^x and/or R^y may not be present) substiutents, which may be attached to any one of the four (in the case of R^x) or three (in the case of R^y) free positions of the benzene ring of the requisite isochromenone bicycle or the requisite phenyl ring bearing the substituents -OR² and -OR³ ring, to which they are attached. For the avoidance of doubt, the identities of each of the optional R^x and R^y substituents are independent. Accordingly, compounds of formula I may be depicted as follows:

in which R¹, R² and R³ are as hereinbefore defined, R^x1, R^x2, R^x3 and R^x4 each independently represent hydrogen or R^x as hereinbefore defined, and R^y1, R*² and R^y3 each independently represent hydrogen or R^y as hereinbefore defined.

Where an alkyl group is terminally substituted with two substituents as hereinbefore defined (e.g. when R², R³ or R^4b are substituted at the terminal position with two A¹ or A⁵ substituents), then those substituents are preferably not both =0 and -OR^6b (for instance when the alkyl group is methyl, so forming a -C(=O)OR^6b group).

Molecular structures may be depicted herein through the use of parentheses to indicate the atom to which a substituent is attached. The substituent in parentheses immediately following an atom is attached via a direct bond to the atom. For example, -C(OR^4a)(R^4b)₂ refers to the following molecular structure:

where * indicates the bond attaching the molecular structure to the rest of the molecule.

For the avoidance of doubt, when a term such as "R^6a to R^6p" is employed herein, this will be understood by the skilled person to mean R^6a, R^6b, R^6c, R^6d, R^6e, R^6f, R⁶⁹, R^6h, R⁶ⁱ, R⁶', R⁶\ R^6m, R⁶ⁿ and R^6p inclusively.

Preferred compounds of the invention include those in which: R² represents hydrogen, Ci-₁₂ alkyl (optionally substituted by one or more substituents selected from A¹), aryl (optionally substituted by one or more substituents selected from A²), heteroaryl (optionally substituted by one or more substituents selected from A³), heterocycloalkyl (optionally substituted by one or more substituents selected from A⁴), -C(O)OR⁵³ or -C(O)N(R^5b)R^5c;

R³ represents hydrogen, C_M2 alkyl (optionally substituted by one or more substituents selected from A¹), -C(O)OR^5a, -C(O)N(R^5b)R^5c or -S(O)₂R^5d; when R¹ represents or -Si(R^6p)₃₎ then each R^6p independently represents C_1-4 alkyl (preferably substituted; e.g. methyl or tert-butyl, so forming for example a -Si(f-butyl)(CH₃)₂ group, i.e. a TBS group).

Further preferred compounds of the invention include those in which: A¹ to A⁸ independently represent aryl (optionally substituted by one or more B¹ substituents), heteroaryl (optionally substituted by one or more substituents selected from B²), heterocycloalkyl (optionally substituted by one or more substituents selected from B³), halo, -CN, -NO₂, -C(O)OR^6a, -O-R^6b, -N(R^7b)R^8b or -C(O)N(R^7a)R^8a; p5b p5c p6a p6b p6d p6f p6h p6i p6j p₆m p6n p₆p p6q p7a p7b p7c p7d p7e p8a p^βb p^βc p^βd _anc| p^βe j_ncj_ep_enc|_entiy represent, on each occasion when used herein, heteroaryl (optionally substituted by one or more substituents selected from B⁶) or, preferably, hydrogen, Ci_-6 alkyl (optionally substituted by one or more substituents selected from B⁴) or aryl (optionally substituted by one or more substituents selected from B⁵); or any pair of R^5b and R^5c, R^7a and R^8a, R^7b and R^8b, R^7c and R^8c, R^7d and R^8d and R^7e and R^8e may be linked together to form, together with the nitrogen atom to which they are necessarily attached, a 3- to 8- (e.g. 5- or 6-) membered ring, optionally containing a further heteroatom (e.g. nitrogen or oxygen), which ring is preferably saturated, and is optionally substituted by one or two (e.g. one) substituent(s) selected from B⁸; R^5a, R^5d, R^6c, R^6e, R⁶⁹, R^6k and R^6r independently represent, on each occasion when used herein, heteroaryl (optionally substituted by one or more substituents selected from B⁶) or, preferably, C_1-6 alkyl (optionally substituted by one or more substituents selected from B⁴) or aryl (optionally substituted by one or more substituents selected from B⁵);

B¹, B², B³, B⁴, B⁵, B⁶, B⁷ and B⁸ independently represent -N(R^9b)R^10b, -NO₂, -CN₁ heteroaryl (optionally substituted by one or more substituents selected from E²) or, preferably, halo, C_1-6 alkyl (optionally substituted by one or more halo atoms) and/or aryl (optionally substituted by one or more substituents selected from E¹); and/or B³, B⁴, B⁷ and B⁸ may alternatively and independently represent =0; R^9a, R^9b and R^1Ob independently represent hydrogen or C_1-3 (e.g. C_1-2) alkyl optionally substituted by one or substituents selected from E⁴.

Preferred compounds of the invention include those in which:

R¹ represents hydrogen or -C(OR^4a)(R^4b)₂;

R² represents heterocycloalkyl (such as tetrahydrofuranyl, e.g. 3-tetrahydrofuranyl; and which heterocycloalkyl group is optionally substituted by one or more substituents selected from A⁴) or, preferably, R² represents hydrogen or C_1-12 alkyl (optionally substituted by one or more substituents selected from A¹);

R³ represents heterocycloalkyl (such as tetrahydrofuranyl, e.g. 3-tetrahydrofuranyl; and which heterocycloalkyl group is optionally substituted by one or more substituents selected from A⁴) or, preferably, R³ represents hydrogen, C_1-12 alkyl (optionally substituted by one or more substituents selected from A¹) or -S(O)₂R^5d; when R² represents optionally substituted C_1-12 alkyl, then such a group may be straight-chain (acyclic) or cyclic, and preferably represents C_1-6 alkyl (e.g. cyclopentyl, butyl (e.g. n-butyl), propyl (e.g. n-propyl), ethyl (which group, when substituted may be substituted at the 1- or 2-position, i.e. forming a -CH₂-CH₂- or -CH(CH₃)- group) or methyl), which alkyl group is optionally substituted by one or more (e.g. two or, preferably, one) substituent(s) selected from A¹; when R³ represents optionally substituted C_1-12 alkyl, then such a group may be straight-chain, cyclic or part-cyclic, and preferably represents C_1-6 alkyl (e.g. cyclopentyl, butyl (e.g. isobutyl, n-butyl or s-butyl), -CH₂-cyclopropyl, ethyl (e.g. which, when substituted, preferably represents -CH₂-CH₂-) or methyl), which alkyl group is optionally substituted by one or more (e.g. one to three) substituent(s) selected from A¹ (however, when A¹ is other than fluoro, then the relevant alkyl group is preferably substituted by two or, more preferably, one, A¹ substituent); for example when R² represents C_1-12 alkyl optionally substituted by one or more A¹ substituents, then:

A¹ preferably represents halo (e.g. fluoro; and hence R² may represent for example, a difluoromethyl or, preferably a trifluoromethyl group) or A¹ preferably represents aryl (such as phenyl; and which aryl group is optionally substituted by one or more B¹ substituents), heteroaryl (such as pyridyl, e.g. 4-pyridyl; and which heteroaryl group is optionally substituted by one or more substituents selected from B²), -C(O)OR^6a or -C(O)N(R^7a)R^8a; for example when R³ represents C_1-12 alkyl optionally substituted by one or more A¹ substituents, then:

A¹ preferably represents aryl (such as phenyl; and which aryl group is optionally substituted by one or more B¹ substituents) halo (e.g. fluoro; and hence R² may represent for example, a difluoromethyl or, preferably a trifluoromethyl group),

-C(O)OR⁶³, -O-R^6b or -C(O)N(R^7a)R^8a;

R^4a represents C_1-3 (e.g. C_1-2) alkyl or, preferably, hydrogen; each R^4b independently represents hydrogen or, preferably, C_1-6 (e.g. C_1-4) alkyl optionally substituted by one or more substituents selected from A⁵ (such groups may for example represent methyl, ethyl or n-propyl); or two R^4b groups when attached to a common carbon atom are linked together to form a

3- to 8- (e.g. 5- or 6-) membered ring, optionally containing one heteroatom (e.g. nitrogen or, preferably, oxygen), which ring is preferably saturated and may be substituted (as hereinbefore defined, i.e. by one or more A⁶ substituents) but is preferably unsubstituted (rings so formed may for example represent cyclohexyl, cyclopentyl or tetrahydropyranyl, e.g. tetrahyadropyran-4-yl);

R^5d represents aryl (e.g. phenyl) optionally substituted by one or more (e.g. one) substituent(s) selected from B⁵; R^6a, R^7a and R^8a independently represent hydrogen;

R^6b represents hydrogen or C₁^ alkyl optionally substituted by one or more substituents selected from B⁴;

B⁴ represents aryl (e.g. phenyl) optionally substituted by one or more substituents selected from E¹; B⁵ represents C_1-2 alkyl (e.g. methyl);

E¹, E², E³ and E⁴ (e.g. E¹) independently represent halo (e.g. fluoro) or C_1-2 alkyl (e.g. methyl).

Further preferred compounds of the invention that may be mentioned include those in which:

R¹ represents hydrogen, 2-hydroxypropan-2-yl, 1-hydroxycyclohexyl, 2-hydroxybutan-

2-yl, 4-hydroxytetrahydropyran-4-yl, 2-hydroxypentan-2-yl, 1-hydroxycyclopentyl or 3- hyd roxypentan-3-yl ;

R² represents hydrogen, cyclopentyl, benzyl, n-butyl, -CH₂-C(O)NH₂, -CH(CH₃)-C(O)NH₂, ethyl, -CH₂-(4-pyridyl), -CH₂C(O)OH, -CH₂-CH₂-C(O)NH₂, methyl and n-propyl;

R³ represents hydrogen, methyl, benzyl, -CH₂-cyclopropyl, isobutyl (i.e.

-CH₂-CH(CHs)₂), n-butyl, s-butyl, cyclopentyl, -CH₂-C(O)NH₂, -CH₂C(O)OH,

-CH₂-CH₂-O-CH₂-phenyl, -CH₂-CH₂-OH and -S(O)₂-(4-methylphenyl); n and m independently represent 2, preferably, 1 or, more preferably, O (i.e. R^x1, R"²,

R^x3, R^x4, R^y1, R^ and R^y3 independently represent hydrogen).

Particularly preferred compounds of the invention include those of the examples described hereinafter.

Compounds of the invention may be made in accordance with techniques that are well known to those skilled in the art, for example as described hereinafter.

According to a further aspect of the invention there is provided a process for the preparation of a compound of formula I which process comprises:

(i) reaction of a compound of formula II,

wherein R , R , R³, R^y and m are as hereinbefore defined, with a compound of formula

wherein L¹ represents a suitable leaving group (e.g. an appropriate halo group, such as iodo), R^s1 represents C_1-6 alkyl (e.g. C_1-3 alkyl such as methyl) optionally substituted by one or more halo atoms, and R^x and n are as hereinbefore defined, for example under coupling reaction conditions, e.g. optionally in the presence of an appropriate metal catalyst (or a salt or complex thereof) such as Cu, Cu(OAc)₂, CuI (or Cul/diamine complex), Pd(OAc)₂, Pd₂(dba)₃ or NiCI₂ and an optional additive such as Ph₃P, 2,2'- bis(diphenylphosphino)-1 ,1'-binaphthyl, xantphos, NaI, LiCI or an appropriate crown ether such as 18-crown-6-benzene, in the presence of an appropriate base such as NaH, Et₃N, pyridine, /V,/V-dimethylethylenediamine, Na₂CO₃, K₂CO₃, DABCO, K₃PO₄, Cs₂CO₃, f-BuONa or f-BuOK (or a mixture thereof), in a suitable solvent (e.g. dichloromethane, dioxane, toluene, ethanol, isopropanol, dimethylformamide, ethylene glycol, ethylene glycol dimethyl ether, water, dimethylsulfoxide, acetonitrile, dimethylacetamide, Λ/-methylpyrrolidinone, tetrahydrofuran or a mixture thereof) or in the absence of an additional solvent when the reagent may itself act as a solvent. This reaction may be carried out at room temperature or above (e.g. at a high temperature, such as 10O⁰C or the reflux temperature of the solvent system that is employed) or using microwave irradiation;

(ii) for compounds of formula I in which R¹ represents hydrogen, de-silylation of a compound corresponding to a compound of formula I but in which R¹ represents -Si(R'¹ )₃, and each R^t1 independently represents a C_1-6 alkyl group (and hence R¹ may represent e.g. terf-butyldimethylsilyl, i.e. -Si(CH₃)f-butyl);

(iii) for compounds of formula I in which R² and/or R³ are other than hydrogen, reaction of a corresponding compound of formula I in which R² and/or R³ (as appropriate) do represent hydrogen, with a compound of formula IV,

R^2/3-L^x IV

wherein L^x represents a suitable leaving group, such as chloro, bromo, iodo or a sulfonate group (e.g. -OS(O)₂CF₃, -OS(O)₂CH₃, -OS(O)₂PhMe or a nonaflate), and R²⁷³ represents either R² or R³ (as appropriate), provided that they do not represent hydrogen, under standard reaction conditions known to those skilled in the art, for example in the presence of a suitable and/or solvent such as those bases and solvents described hereinbefore in respect of process step (i) above; (iv) for compounds of formula I in which R² and/or R³ are hydrogen, reaction of a corresponding compound of formula I in which R² and/or R³ (as appropriate) do not represent hydrogen (e.g. represent C_M2 alkyl optionally substituted by one or more A¹ substituents), with, for example when R² and/or R³ represents alkyl, a strong acid such as HBr and BF₃ or a thiolate anion (e.g. NaSCH₂CH₃) or, for example when R² and/or R³ represent benzyl, by hydrogenolysis under standard conditions.

Compounds of formula Il in which R¹ represents -C(OR^4a)(R^4b)₂ and R^4a represents hydrogen may be prepared by reaction of a corresponding compound of formula Il in which R¹ represents hydrogen with a compound of formula V,

O=C(R^4D)₂ V

wherein R^4b is as hereinbefore defined, for example in the presence of a suitable base or mixtures of bases, for instance sodium hydride, sodium bicarbonate, potassium carbonate, pyrrolidinopyridine, pyridine, triethylamine, diisopropylethylamine, dimethylaminopyridine, diisopropylamine, 1 ,8-diazabicyclo[5.4.0]undec-7-ene, sodium hydroxide, Λ/-ethyldiisopropylamine, Λ/-(methylpolystyrene)-4-(methylamino)pyridine, or preferably with an organometallic base, such as an organolithium base, e.g. methyl lithium, n-BuLi, s-BuLi, f-BuLi, lithium diisopropylamide or lithium 2,2,6,6- tetramethylpiperidine (which organolithium base is optionally in the presence of an additive (for example, a lithium co-ordinating agent such as an ether (e.g. dimethoxyethane) or an amine (e.g. tetramethylethylenediamine (TMEDA), (-)sparteine or 1 ,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone (DMPU) and the like)), for example in the presence of a suitable solvent, such as (in the case where an organometallic base is employed) a polar aprotic solvent (e.g. tetrahydrofuran or diethyl ether), at sub-ambient temperatures (e.g. 0⁰C to -78°C) under an inert atmosphere. The skilled person will appreciate that to obtain such compounds in which R^4a represents hydrogen, the above reaction, when performed in the presence of an organometallic base, will need to be quenched with a suitable proton source, such as H₂O or an aqueous solution of saturated ammonium chloride. The skilled person will also appreciate that the lithium of the lithiated species of the compound of formula III, that may be formed by reaction of a compound of formula III with an organolithium base, may be exchanged to a different metal (i.e. a transmetallation reaction may be performed), for example to zinc (e.g. using ZnCI₂) and the intermediate so formed may then be subjected to reaction with a compound of formula IV₁ for example under reaction conditions described above.

Compounds of formula Il in which R¹ represents -C(R^4b)₃ may be prepared by reaction of a corresponding compound of formula Il in which R¹ represents hydrogen, with a compound of formula Vl,

L²-C(R^4b)₃ Vl

wherein L² represents a suitable leaving group, such as chloro, bromo, iodo or a sulfonate group (e.g. -OS(O)₂CF₃, -OS(O)₂CH₃, -OS(O)₂PhMe or a nonaflate), for example under reaction conditions such as those described above in respect of preparation of compounds of formula Il by reaction of a compound of formula I in which R¹ represents hydrogen and a compound of formula V.

Compounds of formula Il in which R¹ represents hydrogen may be prepared by reaction of a corresponding compound of formula VII,

or a protected derivative thereof, wherein R², R³, R^y and m are as hereinbefore defined, with (Ph₃PCHBr₂)Br (or the like), under standard reaction conditions known to those skilled in the art, for example under Wittig reaction conditions, for example in the presence of a suitable solvent (e.g. tetrahydrofuran) and an appropriate base (e.g. potassium te/t-butoxide or the like) under an inert atmosphere. Alternative reagents include reaction in the presence of (EtO)₂P(O)CH₃ or (Ph)₃P=CHX^2b, for example under standard Homer-Wadsworth-Emmons, or Wittig, reaction conditions, respectively. The skilled person will appreciate that for the synthesis of compounds of formula Il in which R¹ is other than hydrogen, the reagents may be altered accordingly. Compounds of formulae III, IV, V, Vl, VII (and also e.g. certain compounds of formula II) may be commercially available, are known in the literature, or may be obtained either by analogy with the processes described herein, or by conventional synthetic procedures, in accordance with standard techniques, from available starting materials using appropriate reagents and reaction conditions. In this respect, the skilled person may refer to inter alia "Comprehensive Organic Synthesis" by B. M. Trost and I. Fleming, Pergamon Press, 1991.

For convenience only, the compounds illustrated herein are depicted as racemates. One of ordinary skill in the art would recognize that individual enantiomers of the compounds of the invention can be similarly prepared according to the methods disclosed herein, for example using the appropriate chiral starting materials and reagents.

The substituents R¹, R², R³, R^x and R^y either in final compounds of the invention or in relevant intermediates (as appropriate) may be modified one or more times, after or during the processes described above by way of methods that are well known to those skilled in the art. Examples of such methods include substitutions, reductions, oxidations, alkylations, acylations, hydrolyses, esterifications, etherifications, halogenations or nitrations. In this respect, the skilled person may also refer to "Comprehensive Organic Functional Group Transformations" by A. R. Katritzky, O. Meth-Cohn and C. W. Rees, Pergamon Press, 1995. Specific transformation steps that may be mentioned include the conversion of one L¹ group (in the compound of formula II) into another L¹ group (e.g. the conversion of one halo group, such as chloro, into another halo group, such as iodo, for example by reaction in the presence of potassium iodide), or even the conversion of a hydroxy group to a L¹ group. Other transformation steps include the reduction of a nitro group to an amino group, the hydrolysis of a nitrile group to a carboxylic acid group, and standard nucleophilic aromatic substitution reactions.

Compounds of the invention may be isolated from their reaction mixtures using conventional techniques (e.g. recrystallisations). It will be appreciated by those skilled in the art that, in the processes described above and hereinafter, the functional groups of intermediate compounds may need to be protected by protecting groups. The protection and deprotection of functional groups may take place before or after a reaction in the above-mentioned schemes. Functional groups that may need to be protected by suitable protecting groups include hydroxy, amino, mercapto and carboxylic acid. Suitable protecting groups for hydroxy include trialkylsilyl or diarylalkylsilyl (e.g., t-butyldimethylsilyl, t-butyldiphenylsilyl or trimethylsilyl), tetrahydropyranyl, benzyl, methyl and the like. Suitable protecting groups for amino, amidino and guanidino include t-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitable protecting groups for mercapto include -C(O)-R" (where R" is alkyl, aryl or aralkyl), p-methoxybenzyl, trityl and the like. Suitable protecting groups for carboxylic acid include alkyl, aryl or aralkyl esters.

Protecting groups may be added or removed in accordance with standard techniques (for example a methyl protecting group on a hydroxy group may be removed by reaction in the presence of a suitable 'cleaving reagent' such as BBr₃, or a hydroxy group protected with a benzyl group may be deprotected by hydrogenation in the presence of a suitable catalyst such as Pd/C), which are known to one skilled in the art and as described herein. The use of protecting groups is described in detail in Green, T.W. and P.G.M. Wuts, Protective Groups in Organic Synthesis (1999), 3rd Ed., Wiley.

The protecting group may also be a polymer resin such as a Wang resin or a 2- chlorotrityl-chloride resin.

Medical and Pharmaceutical Uses

Compounds of the invention are indicated as pharmaceuticals. According to a further aspect of the invention there is provided a compound of the invention, as hereinbefore defined, for use as a pharmaceutical.

Although compounds of the invention may possess pharmacological activity as such, certain pharmaceutically-acceptable (e.g. "protected") derivatives of compounds of the invention may exist or be prepared which may not possess such activity, but may be administered parenterally or orally and thereafter be metabolised in the body to form compounds of the invention. Such compounds (which may possess some pharmacological activity, provided that such activity is appreciably lower than that of the "active" compounds to which they are metabolised) may therefore be described as "prodrugs" of compounds of the invention.

By "prodrug of a compound of the invention", we include compounds that form a compound of the invention, in an experimentally-detectable amount, within a predetermined time (e.g. about 1 hour), following oral or parenteral administration. All prodrugs of the compounds of the invention are included within the scope of the invention.

Furthermore, certain compounds of the invention may possess no or minimal pharmacological activity as such, but may be administered parenterally or orally, and thereafter be metabolised in the body to form compounds (e.g. compounds of the invention) that possess pharmacological activity as such. Such compounds (which also includes compounds that may possess some pharmacological activity, but that activity is appreciably lower than that of the "active" compounds of the invention to which they are metabolised), may also be described as "prodrugs".

Thus, the compounds of the invention are useful because they possess pharmacological activity, and/or are metabolised in the body following oral or parenteral administration to form compounds which possess pharmacological activity.

The invention disclosed herein is also meant to encompass all pharmaceutically acceptable compounds of the invention being isotopically-labelled by having one or more atoms replaced by an atom having a different atomic mass or mass number. Further, substitution with positron emitting isotopes can be useful in Positron Emission Topography (PET) studies. Isotopically-labelled compounds of the invention can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the Examples and Preparations as set out below using an appropriate isotopically-labelled reagent in place of the non-labelled reagent previously employed.

Compounds of the invention may be useful in treating or preventing inflammatory diseases or conditions in a patient. Hence, in another aspect, this invention is directed to methods for treating or preventing an inflammatory disease or condition in a mammal, preferably a human, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a compound of the invention as hereinbefore described or a therapeutically effective amount of a pharmaceutical formulation/composition of the invention as hereinbefore described.

The inflammatory condition or disease may be an autoimmune condition or disease; the inflammatory condition or disease may involve acute or chronic inflammation of bone and/or cartilage compartments of joints; the inflammatory condition or disease may be an arthritis selected from rheumatoid arthritis, gouty arthritis or juvenile rheumatoid arthritis; the inflammatory condition or disease may be a respiratory disorder selected from asthma or a chronic obstructive pulmonary disease (COPD, e.g., emphysema or chronic bronchitis); the condition or disease may be associated with the disregulation of T-cells; the condition or disease may be associated with elevated levels of inflammatory cytokines (e.g., wherein the inflammatory cytokine is IL-2, or wherein the inflammatory cytokine is IFN-γ, or wherein the inflammatory cytokine is TNF-α); the inflammatory condition or disease may be multiple sclerosis; the inflammatory condition or disease may be pulmonary sarcadosis.; the inflammatory condition or disease may be ocular inflammation or allergy; the inflammatory condition or disease may be an inflammatory bowel disease (e.g., Crohn's disease or ulcerative colitis); and the inflammatory condition or disease may be an inflammatory cutaneous disease (e.g., psoriasis or dermatitis).

Compounds of the invention may be useful in modulating intracellular cyclic adenosine 5'-monophosphate levels within a mammal, preferably a human, Hence, in another aspect, this invention is directed to methods for modulating intracellular cyclic adenosine 5'-monophosphate levels within a mammal, preferably a human, wherein the method comprises administering to the mammal in need thereof an amount of a compound of the invention or a pharmaceutical formulation/composition of the invention as hereinbefore described effective to modulate the intracellular cyclic adenosine 5'-monophosphate levels of the mammal. The mammal, preferably a human, may have an inflammatory condition or disease (for example one defined herein).

Compounds of the invention may be useful in treating or preventing a disease or condition in a mammal, preferably a human, where the disease or condition is associated with pathological conditions that are modulated by inhibiting enzymes associated with secondary cellular messengers. Hence, in another aspect, this invention is directed to methods for treating or preventing a disease or condition in a mammal, preferably a human, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount of a compound of the invention or a pharmaceutical formulation/composition of the invention as hereinbefore described, and the disease or condition is associated with pathological conditions that are modulated by inhibiting enzymes associated with secondary cellular messengers. Such enzymes (that may be inhibited) may be a cyclic AMP PDE; a PDE4; a PDE3; a PDE7; or a cyclic GMP PDE. Further, more than one type of enzyme may be inhibited, for instance, the enzymes may be PDE4, PDE3 and PDE7. The compounds of the invention are preferably inhibitors of PDE4 and, in particular, inhibitors of PDE7, or, dual inhibitors of PDE4 and PDE7.

Compounds of the invention may be useful in treating or preventing uncontrolled cellular proliferation in a mammal, preferably a human. Hence, in another aspect, this invention is directed to methods for treating or preventing uncontrolled cellular proliferation in a mammal, preferably a human, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount (e.g. an amount effective to treat or prevent uncontrolled cellular) of a compound of the invention or a pharmaceutical formulation/composition of the invention as hereinbefore described. The uncontrolled cellular proliferation may be caused by a cancer selected from leukaemia and solid tumors.

Compounds of the invention may be useful in treating or preventing transplant rejection in a mammal, preferably a human. Hence, in another aspect, this invention is directed to methods for treating or preventing transplant rejection in a mammal, preferably a human, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount (e.g. an amount effective to treat or prevent transplant rejection in the mammal) of a compound of the invention or a pharmaceutical formulation/composition of the invention as hereinbefore described. The rejection may be due to graft versus host disease.

Compounds of the invention may be useful in treating or preventing conditions associated with the central nervous system (CNS) in a mammal, preferably a human. Hence, in another aspect, this invention is directed to methods for treating or preventing conditions associated with the central nervous system in a mammal, preferably a human, wherein the method comprises administering to the mammal in need thereof a therapeutically effective amount (e.g. an amount effective to treat or prevent conditions associated with the central nervous system (CNS) in the mammal) of a compound of the invention as described above or a pharmaceutical formulation/composition of the invention as hereinbefore described. The condition associated with the central nervous system (CNS) may be depression.

"Therapeutically effective amount" refers to that amount of a compound of the invention which, when administered to a mammal, preferably a human, is sufficient to effect treatment, as defined below, of a disease or condition of interest in the mammal, preferably a human. The effect may be objective (i.e. measurable by some test or marker) or subjective (i.e. the subject gives an indication of or feels an effect).

The amount of a compound of the invention which constitutes a "therapeutically effective amount" will vary depending on several factors including the compound, the condition and its severity, the manner of administration, and the type of mammal to be treated (e.g. the amount may vary depending on the species, age, weight, sex, renal function, hepatic function and response of the mammal), but can be determined routinely by one of ordinary skill in the art having regard to his own knowledge and to this disclosure.

"Treating" or "treatment" as used herein refers to the therapeutic treatment and/or prophylactic treatment of the disease or condition of interest in a mammal, preferably a human, having the disease or condition of interest. Such terms therefore include: (i) preventing the disease or condition from occurring in a mammal, in particular, when such mammal is predisposed to the condition but has not yet been diagnosed as having it (e.g. prophylactic treatment); or (ii) therapeutic treatment, i.e. treatment of the disease itself, (e.g. complete or partial treatment), which includes:

(a) inhibiting the disease or condition, i.e., arresting its development;

(b) relieving the disease or condition, i.e., causing regression of the disease or condition; or (c) relieving the symptoms resulting from the disease or condition, e.g., relieving swelling without addressing the underlying disease or condition.

As used herein, the terms "disease" and "condition" may be used interchangeably or may be different in that the particular malady or condition may not have a known causative agent (so that etiology has not yet been worked out) and it is therefore not yet recognised as a disease but only as an undesirable condition or syndrome, wherein a more-or-less specific set of symptoms have been identified by clinicians.

Thus, the compounds and compositions of the invention may be used to treat inflammation, including both acute and chronic inflammation as well as certain proliferative disorders (cancers). As used herein, inflammation includes, without limitation, ankylosing spondylitis, arthritis (e.g. juvenile arthritis and rheumatoid arthritis), asthma, COPD, chronic bronchitis, respiratory distress syndrome, rhinitis, allergic rhinitis, Crohn's disease, nephritis, eczema, dermatitis (e.g. atopic dermatitis), urticaria, conjunctivitis, ulcerative colitis, rheumatoid arthritis, osteoarthritis, eosinophilic gastrointestinal disorders, vascular disease, diabetes mellitus, fibromyalgia syndrome, gout, inflammations of the brain (including multiple sclerosis, AIDS dementia, Lyme encephalopathy, herpes encephalitis, Creutzfeld-Jakob disease, and cerebral toxoplasmosis), emphysema, inflammatory bowel disease, irritable bowel syndrome, ischemia-reperfusion injury juvenile erythematosus pulmonary sarcoidosis, Kawasaki disease, osteoarthritis, pelvic inflammatory disease, psoriatic arthritis, psoriasis, tissue/organ transplant, scleroderma, spondyloarthropathies, systemic lupus erythematosus, pulmonary sarcoidosis, ulcerative colitis, viral infections (i.e. inflammation associated with a viral infection) (e.g. influenza, common cold, herpes zoster, hepatitis C and AIDS), bacterial infections (i.e. inflammation associated with a bacterial infection), and any other disease with an inflammatory component. As used herein, proliferative disorders includes, without limitation, all cancers, leukemias and solid tumors that are susceptible to undergoing differentiation or apoptosis upon interruption of their cell cycle. As stated herein, the compounds and compositions of the invention may also be useful for treating diseases associated with the central nervous system. Such diseases include cognitive function, Alzheimer's disease and other neurodegenerative disorders, learning and memory disorders. The term "inflammation" will be understood by those skilled in the art to include any condition characterised by a localised or a systemic protective response, which may be elicited by physical trauma, infection, chronic diseases, such as those mentioned hereinbefore, and/or chemical and/or physiological reactions to external stimuli (e.g. as part of an allergic response). Any such response, which may serve to destroy, dilute or sequester both the injurious agent and the injured tissue, may be manifest by, for example, heat, swelling, pain, redness, dilation of blood vessels and/or increased blood flow, invasion of the affected area by white blood cells, loss of function and/or any other symptoms known to be associated with inflammatory conditions.

The term "inflammation" will thus also be understood to include any inflammatory disease, disorder or condition per se, any condition that has an inflammatory component associated with it, and/or any condition characterised by inflammation as a symptom, including inter alia acute, chronic, ulcerative, specific, allergic and necrotic inflammation, and other forms of inflammation known to those skilled in the art. The term thus also includes, for the purposes of this invention, inflammatory pain and pain generally.

Where a condition has an inflammatory component associated with it, or a condition characterised by inflammation as a symptom, the skilled person will appreciate that compounds of the invention may be useful in the treatment of the inflammatory symptoms and/or the inflammation associated with the condition.

Compounds of the invention may inhibit disease induction in these models at doses of less than 20 mg/kg. The Biological Examples below outline some, but not all, of the preclinical models that may be used to support the claims of this patent.

Administration of the compounds of the invention, or their pharmaceutically acceptable salts, in pure form or in an appropriate pharmaceutical composition, can be carried out via any of the accepted modes of administration of agents for serving similar utilities.

For instance, compounds of the invention may be administered orally, intravenously, subcutaneously, buccally, topically, vaginally, rectally, dermally, transdermal^, nasally, tracheally, bronchially, sublingually, by any other parenteral route or via inhalation, in a pharmaceutically acceptable dosage form. The term parenteral as used herein includes subcutaneous injections, intravenous, intramuscular, intrastemal injection or infusion techniques. Current methods for ocular delivery include topical administration (eye drops), subconjunctival injections, periocular injections, intravitreal injections, surgical implants and iontophoresis. Those skilled in the art would combine the best suited excipients with the compound for safe and effective intra-ocular administration.

Compounds of the invention may be administered alone, but are preferably administered by way of known pharmaceutical compositions/formulations, including tablets, capsules or elixirs for oral administration, suppositories for rectal administration, sterile solutions or suspensions for parenteral or intramuscular administration, and the like.

According to a further aspect of the invention there is provided a pharmaceutical composition/formulation including a compound of the invention, as hereinbefore defined, in admixture with a pharmaceutically acceptable adjuvant, carrier, diluent or excipient.

Depending on e.g. potency and physical characteristics of the compound of the invention (i.e. active ingredient), pharmaceutical formulations that may be mentioned include those in which the active ingredient is present in at least 1% (or at least 10%, at least 30% or at least 50%) by weight. That is, the ratio of active ingredient to the other components (i.e. the addition of adjuvant, diluent and carrier) of the pharmaceutical composition is at least 1 :99 (or at least 10:90, at least 30:70 or at least 50:50) by weight.

Such compositions/formulations may be prepared in accordance with standard and/or accepted pharmaceutical practice. For example, pharmaceutical compositions of the invention may be formulated so as to allow the active ingredients contained therein to be bioavailable upon administration of the composition to a patient. Compositions that will be administered to a subject or patient take the form of one or more dosage units, where for example, a tablet may be a single dosage unit, and a container of a compound of the invention in aerosol form may hold a plurality of dosage units. Actual methods of preparing such dosage forms are known, or will be apparent, to those skilled in this art; for example, see The Science and Practice of Pharmacy, 20th Edition (Philadelphia College of Pharmacy and Science, 2000). The composition to be administered will, in any event, contain a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, for treatment of a disease or condition of interest in accordance with the teachings of this invention.

The invention further provides a process for the preparation of a pharmaceutical composition/formulation, as hereinbefore defined, which process comprises bringing into association a compound of the invention, as hereinbefore defined, or a pharmaceutically acceptable derivative (e.g. salt) thereof, with a pharmaceutically- acceptable adjuvant, carrier, diluent or excipient.

Compounds of the invention may also be combined with other therapeutic agents that are useful in the treatment of the conditions described herein. For instance, the compounds of the invention may be combined with other compounds that may be useful in the treatment of: i) an inflammatory disorder; ii) a disorder in which the modulation of intracellular cyclic adenosine 5¹- monophosphate levels within a mammal is desired and/or required, which disorder may be an inflammatory disorder; iii) a disorder associated with pathological conditions that are modulated by inhibiting enzymes associated with secondary cellular messengers (e.g. a cyclic AMP PDE; a PDE4; a PDE3; a PDE7; a cyclic GMP PDE; or PDE4, PDE3 and PDE7), which disorder may be an inflammatory disorder (it is most preferred that compounds of the invention are combined (an) inhibitor(s) of PDE4 or, in particular, (an) inhibitor(s) of PDE7, or, dual inhibitors of PDE4 and PDE7); iv) transplant rejection in a mammal; v) uncontrolled cellular proliferation; and/or vi) a disorder associated with the central nervous system.

According to a further aspect of the invention, there is provided a combination product comprising:

(A) a compound of the invention, as hereinbefore defined; and

(B) another therapeutic agent that is useful in the treatment of i), ii), iii), iv), v) or vi) above (e.g. a therapeutic agent that is useful in the treatment of an inflammatory disorder), wherein each of components (A) and (B) is formulated in admixture with a pharmaceutically-acceptable adjuvant, diluent, carrier or excipient.

Such combination products provide for the administration of a compound of the invention in conjunction with the other therapeutic agent, and may thus be presented either as separate formulations, wherein at least one of those formulations comprises a compound of the invention, and at least one comprises the other therapeutic agent, or may be presented (i.e. formulated) as a combined preparation (i.e. presented as a single formulation including a compound of the invention and the other therapeutic agent).

Thus, there is further provided:

(1 ) a pharmaceutical formulation/composition including a compound of the invention, as hereinbefore defined, another therapeutic agent that is useful in the treatment of i), ii), iii), iv), v) or vi) above (e.g. a therapeutic agent that is useful in the treatment of an inflammatory disorder), and a pharmaceutically-acceptable adjuvant, diluent, carrier or excipient; and

(2) a kit of parts comprising components:

(a) a pharmaceutical formulation/composition including a compound of the invention, as hereinbefore defined, in admixture with a pharmaceutically- acceptable adjuvant, diluent, carrier or excipient; and

(b) a pharmaceutical formulation/composition including another therapeutic agent that is useful in the treatment of i), ii), iii), iv), v) or vi) above (e.g. a therapeutic agent that is useful in the treatment of an inflammatory disorder) in admixture with a pharmaceutically-acceptable adjuvant, diluent, carrier or excipient, which components (a) and (b) are each provided in a form that is suitable for administration in conjunction with the other.

The invention further provides a process for the preparation of a combination product as hereinbefore defined, which process comprises bringing into association a compound of the invention, as hereinbefore defined, or a pharmaceutically acceptable derivative (e.g. salt) thereof with another therapeutic agent that is useful in the treatment of i), ii), iii), iv), v) or vi) above (e.g. a therapeutic agent that is useful in the treatment of an inflammatory disorder), and at least one pharmaceutically-acceptable adjuvant, diluent, carrier or excipient.

By "bringing into association", we mean that the two components are rendered suitable for administration in conjunction with each other.

Thus, in relation to the process for the preparation of a kit of parts as hereinbefore defined, by bringing the two components "into association with" each other, we include that the two components of the kit of parts may be:

(i) provided as separate formulations (i.e. independently of one another), which are subsequently brought together for use in conjunction with each other in combination therapy; or (ii) packaged and presented together as separate components of a "combination pack" for use in conjunction with each other in combination therapy.

"Pharmaceutically acceptable salt" includes both acid and base addition salts.

"Pharmaceutically acceptable acid addition salt" refers to those salts which retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and which are formed with organic or inorganic acids.

"Pharmaceutically acceptable base addition salt" refers to those salts which retain the biological effectiveness and properties of the free acids, which are not biologically or otherwise undesirable. These salts are prepared from addition of an inorganic base or an organic base to the free acid.

Often crystallisations produce a solvate of the compound of the invention. As used herein, the term "solvate" refers to an aggregate that comprises one or more molecules of a compound of the invention with one or more molecules of solvent. The solvent may be water, in which case the solvate may be a hydrate. Alternatively, the solvent may be an organic solvent. The compound of the invention may be a true solvate, while in other cases, the compound of the invention may merely retain adventitious water or be a mixture of water plus some adventitious solvent.

The compounds of the invention, or their pharmaceutically acceptable salts, are administered in a therapeutically effective amount, which will vary depending upon a variety of factors including the activity of the specific compound employed; the metabolic stability and length of action of the compound; the age, body weight, general health, sex, and diet of the patient; the mode and time of administration; the rate of excretion; the drug combination; the severity of the particular disease or condition; and the subject undergoing therapy.

Compounds of the invention may be administered at varying doses. Oral, pulmonary and topical dosages may range from between about 0.01 mg/kg of body weight per day (mg/kg/day) to about 100 mg/kg/day, preferably about 0.01 to about 10 mg/kg/day, and more preferably about 0.1 to about 5.0 mg/kg/day. For e.g. oral administration, the compositions typically contain between about 0.01 mg to about 500 mg, and preferably between about 1 mg to about 100 mg, of the active ingredient. Intravenously, the most preferred doses will range from about 0.001 to about 10 mg/kg/hour during constant rate infusion. Advantageously, compounds may be administered in a single daily dose, or the total daily dosage may be administered in divided doses of two, three or four times daily. When a pharmaceutical composition containing a compound of the invention is employed, it shall contain an appropriate amount/concentration/ratio of the active ingredient.

The ranges of effective doses provided herein are not intended to be limiting and represent preferred dose ranges. However, the most preferred dosage will be tailored to the individual subject, as is understood and determinable by one skilled in the relevant arts, (see, e.g., Berkow et al., eds., The Merck Manual, 16^th edition, Merck and Co., Rahway, N.J., 1992; Goodmanetna., eds..Goodman and Cilman's The Pharmacological Basis of Therapeutics, 10^th edition, Pergamon Press, Inc., Elmsford, N.Y., (2001); Avery's Drug Treatment: Principles and Practice of Clinical Pharmacology and Therapeutics, 3rd edition, ADIS Press, LTD., Williams and Wilkins, Baltimore, MD. (1987), Ebadi, Pharmacology, Little, Brown and Co., Boston, (1985); Osolci al., eds., Remington's Pharmaceutical Sciences, 18^th edition, Mack Publishing Co., Easton, PA (1990); Katzung, Basic and Clinical Pharmacology, Appleton and Lange, Norwalk, CT (1992)).

The physician, or the skilled person, will be able to determine the actual dosage and/or route of administration which will be most suitable for an individual patient, which is likely to vary with the route of administration, the type and severity of the condition that is to be treated, as well as the species, age, weight, sex, renal function, hepatic function and response of the particular patient to be treated. The above-mentioned dosages are exemplary of the average case; there can, of course, be individual instances where higher or lower dosage ranges are merited, and such are within the scope of this invention.

Compounds of the invention may have the advantage that they are effective inhibitors of PDE, such as PDE4 and, particularly, PDE7 (or compounds of the invention may be effective dual inhibitors of PDE4 and PDE7).

Compounds of the invention may also have the advantage that they may be more efficacious than, be less toxic than, be longer acting than, be more potent than, produce fewer side effects than, be more easily absorbed than, and/or have a better pharmacokinetic profile (e.g. higher oral bioavailability and/or lower clearance) than, and/or have other useful pharmacological, physical, or chemical properties over, compounds known in the prior art, whether for use in the above-stated indications or otherwise.

Examples

The invention is illustrated by way of the following examples. The following abbreviations may be employed:

aq aqueous

Bn benzyl brine saturated aqueous solution of NaCI

DCM dichloromethane

DMF dimethylformamide

EtOAc ethyl acetate

EtOH ethanol

KOtBu Potassium tert-butoxide

Me methyl

MeCN acetonitrile

MeOH methanol NMR nuclear magnetic resonance

Pd/C palladium on carbon rt room temperature sat saturated TBS tert-butyldimethylsilyl

THF tetrahydrofuran

TLC thin layer chromatography

In general, starting components may be obtained from sources such as Sigma Aldrich, Lancaster Synthesis, Inc., Maybridge, Matrix Scientific, TCI, and Fluorochem USA, etc. or synthesized according to sources known to those skilled in the art (see, e.g., Smith, M. B. and J. March, Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 5th edition (Wiley, December 2000)) or prepared as described herein.

PREPARATION OF COMPOUNDS (2), (3) AND (4)

A. A mixture of 3-hydroxy-4-methoxybenzaldehyde (1 ) (22.5 g, 148 mmol), benzylbromide (35.2 mL, 294 mmol), K₂CO₃ (61.3 g, 444 mmol), Kl (736 mg, 4.4 mmol) and DMF was stirred at 60 ⁰C for 5 d. Standard work-up followed by chromatography gave 31.5 g (88%) of Compound (2).

B. A mixture of Compound (2) (21.94 g, 91 mmol), NaSEt ( 80%, 10 g, 95 mmol) and DMF (150 mL) was stirred at 100 ⁰C for 45 min. The mixture was cooled to rt, quenched with 1 M HCI and extracted with toluene. The combined extracts were washed with sat NH₄CI, dried and concentrated. Purification of the residue by chromatography gave 14.9 g (72.3%) of Compound (3).

C. A mixture of Compound (3) (14.9 g, 65.2 mmol), cyclopentylbromide (13.98 ml, 130.6 mmol), K₂CO₃ (27.0 g, 195.6 mmoL), Kl (325 mg) and DMF (125 mL) was stirred at 60 ⁰C for 3 d. The mixture was cooled to rt, diluted with water and extracted with toluene. The combined extracts were washed with 1 M NaOH, brine , dried over MgSO₄ and filtered. Concentration and purification of the residue by chromatography gave 16.8 g (87%) of Compound (4). PREPARATION OF COMPOUND (5)

Cyclopentylbromide (80 mL, 0.75 mol) was added to a mixture of compound (1 ) (100 g, 657 mmol), Kl (3.3 g, 19.7 mmol), K₂CO₃ (136 g, 986 mmol) and DMF (500 mL) at 65 ⁰C and the mixture was stirred at that temperature for 16 h. Cyclopentylbromide (12 mL, 0.11 mol) was added and the mixture was stirred for 4 d at 65 ⁰C. The mixture was cooled to rt, diluted with water and extracted with toluene. The organic layer was washed with 1 M NaOH and dried over MgSO₄. Concentration gave 142 g (98%) of Compound (5).

PREPARATION OF COMPOUNDS (7-11 )

A. Benzylbromide (16.8 mL, 141.2 mmol) was added to a mixture of 3,4-dihydroxybenzaldehyde (6) (15.0 g, 108.6 mmol), Kl (1.8 g, 10.9 mmol), NaHCO₃

(10.5 g, 124.9 mmol) and MeCN (125 mL) at 40 ⁰C and the mixture was stirred at 40 ⁰C for 2-6 days with monitoring by TLC. The mixture was cooled to rt, 0.1 M HCI was added and the mixture was extracted with toluene. The organic layer was washed with brine and dried over MgSO₄. Concentration gave Compound (7), which was used without further purification.

B. n-Propyl iodide (4.6 mL, 57 mmol) was added to a mixture of 3-hydroxy- 4-benzyloxybenzaldehyde (7) (7.0 g, 30.7 mmol), K₂CO₃ (12.7 g, 92 mmol) and DMF (75 mL) at 65 ⁰C, and the mixture was stirred at 65 °C for 24 h. The mixture was cooled to rt, diluted with sat NaHCO₃ and extracted with diethyl ether. The combined extracts were washed with 1 M NaOH, water, and brine and dried over MgSO₄. Concentration gave 8.28 (99%) of Compound (8). Compounds (9), (10), (11) were prepared using this procedure.

PREPARATION OF COMPOUND (12)

t-Butyldimethylsilylchloride (9.3 g, 61.4 mmol) was added to a mixture of Compound (7) (7.0 g, 30.7 mmol), imidazole (8.4 g, 122.8 mmol) and DMF (150 ml_). The mixture was stirred at rt and monitored by TLC. At or near completion, the mixture was subjected to standard work-up procedures and Compound (12) was isolated.

PREPARATION OF COMPOUND (13)

t-Butyldimethylsilylchloride (1.9 ml_, 7.24 mmol) was added to a mixture of

Compound (6) (1.0 g, 7.24 mmol), imidazole (0.49 g, 7.24 mmol) and DMF (20 mL) at 0 ⁰C. The mixture was warmed to rt and monitored by TLC. At or near completion the mixture was subjected to standard work-up procedures and Compound (13) was isolated. PREPARATION OF COMPOUNDS (14-17)

lsobutyl iodide (2.2 ml_, 18.9 mmol) was added to a mixture of Compound (3) (3.3 g, 14.5 mmol), K₂CO₃ (3.0 g, 21.8 mmol) and DMF (40 mL) at 65 ⁰C. The mixture was stirred at 65 °C while monitoring by TLC. Standard work-up gave Compound (14). Compounds (15), (16), (17) were prepared using this procedure.

PREPARATION OF COMPOUND (18)

(5) (18)

A mixture of (Ph₃PCHBr₂)Br (13.45 g, 26.11 mmol), KOtBu (2.79 g, 24.87 mmol) and THF (120 mL) was stirred at rt for 20 min. Compound (5) (2.94 g, 13.35 mmol) was added. After 10 min at rt, KOtBu (7.33 g, 65.28 mmol) was added and the mixture was stirred at rt for 20 min. Brine was added and the mixture was extracted with diethyl ether. The combined extracts were dried (MgSO₄) and concentrated and the residue purified by chromatography to give 2.80 g (97%) of Compound (18). PREPARATION OF COMPOUND (19)

KOtBu (720 mg, 6.40 mmol) was added to a mixture of PPh₃CHBr₂^Br (3.47 g, 6.74 mmol) and THF (30 ml_), and the mixture was stirred at rt for 10 min. A solution of Compound (4) (1 g, 3.37 mmol) in THF was added over 10 min. KOtBu (1.89 g, 16.9 mmol) was added portionwise over a period of 5 min and the mixture was stirred for 30 min. The mixture was extracted with diethyl ether. The combined extracts were dried (MgSO₄) and concentrated The residue was purified by chromatography to give 0.76 g (77%) of Compound (19).

PREPARATION OF COMPOUND (20)

In a similar manner as described above in the preparation of Compound (19), Compound (20) was prepared from Compound (2). PREPARATION OF COMPOUND (21)

(14) (21)

In accordance with the procedures described herein, Compound (21) was prepared from Compound (14).

PREPARATION OF COMPOUND (22)

(15) (22)

In accordance with the procedures described herein, Compound (22) was prepared from Compound (15).

PREPARATION OF COMPOUND (23)

(16) (23)

In accordance with the procedures described herein, Compound (23) was prepared from Compound (16). PREPARATION OF COMPOUND (24)

In accordance with the procedures described herein, Compound (24) was prepared from Compound (17).

PREPARATION OF COMPOUNDS (25 & 26)

(12) (25) (26)

In accordance with the procedures described herein, Compound (25) and Compound (26) was prepared from Compound (12).

PREPARATION OF COMPOUND (27)

A solution of MeLi (1.4 M in diethyl ether, 6.9 mL, 9.71 mmol) was added to a stirred solution of Compound (18) (2.00 g, 9.25 mmol) in THF (40 mL) at -40 ⁰C. The mixture was allowed to warm to 0 ^CC over 1 h, and acetone (1.4 mL, 18.5 mmol) was added. The mixture was stirred at 0 ⁰C for 1 h, quenched with sat aq NH₄CI, and extracted with EtOAc . The combined extracts were dried (MgSO₄) and concentrated and the residue was purified by chromatography to give 2.25 g (87%) of Compound (27) (2.25 g, 87%).

PREPARATION OF COMPOUND (28)

A solution of MeLi (1.4 M in diethyl ether, 5.2 ml_, 7.29 mmol) was added to a stirred solution of Compound (18) (1.5 g, 6.94 mmol) in THF (20 ml.) at -40 ⁰C. The mixture was allowed to warm to 0 ⁰C over 1 h, and 2-butanone (1.2 ml_, 13.9 mmol) was added. The mixture was stirred at 0 ⁰C for 1 h, quenched with sat aq NH₄CI, and extracted with EtOAc. The combined extracts were dried (MgSO₄) and concentrated and the residue was purified by chromatography to give 1.91 g (96%) of Compound (28).

PREPARATION OF COMPOUND (29)

In accordance with the procedures described herein, Compound (29) was prepared from Compound (18). PREPARATION OF COMPOUND (30)

In accordance with the procedures described herein, Compound (30) was prepared from Compound (18).

PREPARATION OF COMPOUND (31 )

In accordance with the procedures described herein, Compound (31) was prepared from Compound (18).

PREPARATION OF COMPOUND (32)

In accordance with the procedures described herein, Compound (32) was prepared from Compound (24).

PREPARATION OF COMPOUND (33)

(23) (33)

In accordance with the procedures described herein, Compound (33) was prepared from Compound (23).

PREPARATION OF COMPOUND (34)

(21) (34) In accordance with the procedures described herein, Compound (34) was prepared from Compound (21 ).

PREPARATION OF COMPOUND (35)

(22) (35)

In accordance with the procedures described herein, Compound (35) was prepared from Compound (22).

PREPARATION OF COMPOUND (36)

In accordance with the procedures described herein, Compound (36) was prepared from Compound (19).

PREPARATION OF COMPOUND (37)

In accordance with the procedures described herein, Compound (37) was prepared from Compound (25).

PREPARATION OF COMPOUND (38)

(18) (38)

A solution of MeLi (1.4 M in diethyl ether, 59 ml_, 83.22 mmol) was added to a stirred solution of Compound (18) (15.00 g, 69.35 mmol) in THF (50 ml.) at -78 ⁰C. The mixture was allowed to warm up to -10 ⁰C over 1 h. The mixture was cooled to -50 ⁰C and terf-butyldimethylsilyl chloride (13.59 g, 90.16 mmol) was added. The mixture was allowed to warm up to rt and was stirred overnight. Sat aq NaHCO₃ was added, and the mixture was extracted with EtOAc. The combined extracts were dried over MgSO₄ and concentrated. The residue was purified by chromatography to give 18.8 g (82%) of Compound (38). PREPARATION OF COMPOUND (39)

(18) (39)

In accordance with the procedures described herein (e.g. for the synthesis of Compound (38)), Compound (39) was prepared from Compound (18) using methyl chloroformate as the electrophile.

PREPARATION OF COMPOUND (40)

(39) (40)

Compound (40) was prepared from Compound (39) by hydrolysis (LiOH in THF/water).

PREPARATION OF COMPOUND (41 )

In accordance with the procedures described herein, Compound (41 ) was prepared from Compound (18). D. Coupling Reactions - Compounds of Formula (I)

Compounds of formula (I) are prepared by coupling the appropriately substituted phenylalkynyl intermediates (compounds of formula (VII)) with iodobenzoate compounds, as shown below in Reaction Scheme 5, where R¹, R² and R³ are as defined above in the Summary of the Invention, such as compound (55) which is commercially available or which can be prepared from the benzoic acid precursor according to methods known to one skilled in the art. Examples 30-42 show how a number of compounds of formula (I) may be generated using the iodobezoate (55) and various compounds of formula (VII). REACTION SCHEME 5

Iodobenzoate compounds, such as compound (55), are commercially available or can be prepared according to methods known to one skilled in the art from the benzoic acid precursor. The following Synthetic Examples show how a number of compounds of formula (I) may be generated using the iodobezoate (55) and various compounds of formula (VII).

EXAMPLE 1 (COMPOUND 42) 4-(3-Cvclopentyloxy-4-methoxyphenvπ-3-(2-hvdroxy-2-propyl)isochromen-1-one

A mixture of DMF (6OmL) and methyl 2-iodobenzoate (0.59 mL, 4.01 mmol) was added to a mixture of Compound (27) (2.20 g, 8.02 mmol), Pd(OAc)₂ (45 mg, 0.2 mmol), Na₂CO₃ (0.43 g, 4.01 mmol) and LiCI (170 mg, 4.01 mmol) at rt. The mixture was stirred at 100 ⁰C for 20 h and allowed to reach rt. Toluene and water were added, the layers separated and the aq layer extracted with toluene. The combined organic phases were washed with water, brine and dried over MgSO₄. Concentration and purification of the residue by chromatography gave 1.25 g (79%) of the title compound.

EXAMPLE 2 (COMPOUND 43) 4-(3-Cvclopentyloxy-4-methoxyphenyl)-3-(1-hvdroxycvclohexyl)isochromen-1-one

(41) (43) In accordance with the procedure described in Example 1 , the title compound was prepared from Compound (55) and Compound (41 ). EXAMPLE 3 (COMPOUND 44) 4-(3-Cvclopentyloxy-4-methoxyphenyl)-3-(2-hvdroxybutan-2-yl)isochromen-1-one

(28) (44)

In accordance with the procedure described in Example 1, the title compound was prepared from Compound (55) and Compound (28).

EXAMPLE 4 (COMPOUND 45)

4-(3-Cvclopentyloxy-4-methoxyphenyl)-3-(4-hydroxytetrahvdropyran-4-yl)isochromen-

1-one

(55) (30) (45)

In accordance with the procedure described in Example 1 , the title compound was prepared from Compound (55) and Compound (30). EXAMPLE 5 (COMPOUND 47) 4-(3-Cvclopentvloxv-4-methoxvphenvl)-3-(2-hvdroxvpentan-2-vl)isochromen-1-one

(31) (47)

In accordance with the procedure described in Example 1 , the title compound was prepared from Compound (55) and Compound (31 ).

EXAMPLE 6 (COMPOUND 48) S-fte/t-ButyldimethylsilylM-O-cvclopentyloxy^-methoxyphenvDisochromen-i-one

DMF (25 mL) and methyl 2-iodobenzoate (0.317 g, 1.21 mmol) were added to a mixture of Compound (38) (0.80 g, 2.42 mmol), Pd(OAc)₂ (13.6 mg, 0.06 mmol),

Na₂CO₃ (0.128 g, 1.21 mmol) and LiCI (51 mg, 1.21 mmo!) at rt. The mixture was stirred at 10O ⁰C overnight and allowed to reach rt. Toluene and water were added and the mixture was extracted with toluene. The combined organic phases were washed with brine and dried over MgSO₄. Concentration and purification by chromatography followed by crystallization from hexane gave 0.256 g (47%) of the title compound. EXAMPLE 7 (COMPOUND 49) 4-(3.4-Bis(benzyloxy)phenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

In accordance with the procedure described in Example 1 , the title compound was prepared from Compound (55) and Compound (32).

EXAMPLE 8 (COMPOUND 50) 4-(3-Benzyloxy-4-cvclopropylmethoxyphenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

(55) (33) (50)

In accordance with the procedure described in Example 1 , the title compound was prepared from Compound (55) and Compound (33).

EXAMPLE 9 (COMPOUND 51 ) 4-(3-Benzyloxy-4-isobutoxyphenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

In accordance with the procedure described in Example 1 , the title compound was prepared from Compound (55) and Compound (34).

EXAMPLE 10 (COMPOUND 52) 4-(3-Benzyloxy-4-butoxyphenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

(55) (35) (52)

In accordance with the procedure described in Example 1 , the title compound was prepared from Compound (55) and Compound (35).

EXAMPLE 11 (COMPOUND 53) 4-(3-Benzyloxy-4-cvclopentyloxyphenyl)-3-(2-hvdroxy-2-propy0isochromen-1-one

(55) (36) (53)

In accordance with the procedure described in Example 1 , the title compound was prepared from Compound (55) and Compound (36).

EXAMPLE 12 (COMPOUND 56) 4-(3-Cvclopentyloxy-4-methoxyphenyl)isochromen-1-one

A mixture of Compound (48) (0.90 g, 1.77 mmol), H-Bu₄NCI (1.48 g, 5.31 mmol), KF.2H₂O (0.53 g, 5.66 mmol) and MeCN (5 mL) were stirred at rt overnight.

Water was added and the mixture was extracted with EtOAc. The combined extracts were dried over MgSO₄ and concentrated and the residue was purified by chromatography to give 0.59 g (85%) of the title compound EXAMPLE 13 (COMPOUND 60) 4-(3.4-Dihvdroxyphenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

(49) (60)

A mixture of Compound (49) (0.31 g, 6.3 mmol), Pd/C (10%, 0.3 g) and EtOAc (60 ml.) was hydrogenated at ambient temperature and pressure for 48 h. Filtration, concentration and purification by chromatography gave 1.44 g (73%) of the title compound.

EXAMPLE 14 (COMPOUND 61) 4-(4-Cvclopropylmethoxy-3-hvdroxyphenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

(50) (61)

The title compound was prepared from Compound (50) in accordance with the procedure described in Example 13. Yield: 1.20 g (73%). EXAMPLE 15 (COMPOUND 62) 4-(3-Hvdroxy-4-isobutoxyphenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

(51) (62)

The title compound was prepared from Compound (51) in accordance with the procedure described in Example 13. Yield: 0.82g (57%).

EXAMPLE 16 (COMPOUND 63) 4-(4-Butoxy-3-hvdroxyphenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

(52) (63) The title compound was prepared from Compound (52) in accordance with the procedure described in Example 13. Yield: 1.04 g (65%).

EXAMPLE 17 (COMPOUND 64) 4-(4-Cvclopentyloxy-3-hvdroxyphenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

The title compound was prepared from Compound (53) in accordance with the procedure described in Example 13. Yield: 1.1O g (76%).

EXAMPLE 18 (COMPOUND 65) 4-(3,4-Diethoxyphenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

A mixture of Compound (60) (100 mg, 0.32 mmol), ethyl iodide (152 μL, 1.91 mmol), K₂CO₃ (40 mg, 2.8 mmol) and DMF (5 mL) was hetaed at 65 ⁰C for 48 h. Standard work-up and purification by cromatography gave the title compound. Yield: 92 mg (78%). EXAMPLE 19 (COMPOUND 66) 4-(3.4-Dibutoχyphenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

The title compound was prepared from Compound (60) and /7-butyl iodide in accordance with the procedure described in Example 18.

EXAMPLE 20 (COMPOUND 67)

2-(2-Cvclopropylmethoxy-5-(3-(2-hvdroxy-2-propyl)-1-oxoisochromen-4-yl)phenoxy)- acetamide

(61) (67)

The title compound was prepared from Compound (61 ) and iodoacetamide in accordance with the procedure described in Example 18.

EXAMPLE 21 (COMPOUND 68)

2-(2-Cvclopropylmethoxy-5-(3-(2-hvdroxy-2-propy0-1-oxoisochromen-4-yl)phenoxy)- propanamide

The title compound was prepared from Compound (61) and 2-bromo propionamide in accordance with the procedure described in Example 18.

EXAMPLE 22 (COMPOUND 69) 4-(4-Cvclopropylmethoxy-3-ethoxyphenvπ-3-(2-hvdroxy-2-propyl)isochromen-1-one

(61) (69) The title compound was prepared from Compound (61) and ethyl iodide in accordance with the procedure described in Example 18.

EXAMPLE 23 (COMPOUND 79) 2-(5-(3-(2-hvdroxy-2-propyl)-1-oxoisochromen-4-yl)-2-isobutoxyphenoxy)acetamide

(62) (70)

The title compound was prepared from Compound (62) and iodoacetamide in accordance with the procedure described in Example 18.

EXAMPLE 24 (COMPOUND 71 )

2-(5-(3-(2-Hvdroxypropan-2-v0-1-oxoisochromen-4-yl)-2-isobutoxyphenoxy)propan- amide

The title compound was prepared from Compound (62) and 2-bromopropion- amide in accordance with the procedure described in Example 18. EXAMPLE 25 (COMPOUND 72) 4-(3-Ethoxy-4-isobutoxyphenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

(62) (72)

The title compound was prepared from Compound (62) and ethyl iodide in accordance with the procedure described in Example 18.

EXAMPLE 26 (COMPOUND 73) 2-(2-Butoxy-5-(3-(2-hvdroxy-2-propyl)-1-oxoisochromen-4-yl)phenoxy)acetamide

(63) (73)

The title compound was prepared from Compound (63) and iodoacetamide in accordance with the procedure described in Example 18.

EXAMPLE 27 (COMPOUND 74) 2-(2-Butoxy-5-(3-(2-hvdroxy-2-propyl)-1-oxoisochromen-4-yl)phenoxy)propanarnide

(63) (74)

The title compound was prepared from Compound (63) and 2-bromopropion- amide in accordance with the procedure described in Example 18.

EXAMPLE 28 (COMPOUND 75) 4-(4-Butoxy-3-ethoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one

(63) (75)

The title compound was prepared from Compound (63) and ethyl iodide in accordance with the procedure described in Example 18.

EXAMPLE 29 (COMPOUND 76)

2-(2-Cvclopentyloxy-5-(3-(2-hvdroxy-2-propyl)-1-oxoisochromen-4-yl)phenoxy)acet- amide

(64) (76)

The title compound was prepared from Compound (64) and iodoacetamide in accordance with the procedure described in Example 18.

EXAMPLE 30 (COMPOUND 77)

2-(2-Cvclopentyloxy-5-(3-(2-hvdroxy-2-propyl)-1-oxoisochromen-4-yl)phenoxy)- propanamide

(64) (77)

The title compound was prepared from Compound (64) and 2- bromopropionamide in accordance with the procedure described in Example 18. EXAMPLE 31 (COMPOUND 78) 4-(4-Cvclopentyloxy-3-ethoxyphenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

(64) (78)

The title compound was prepared from Compound (64) and ethyl iodide in accordance with the procedure described in Example 18.

EXAMPLE 32 (COMPOUND 79) 4-(3-Butoxy-4-cvclopentyloxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one

(64) (79)

The title compound was prepared from Compound (64) and n-butyl iodide in accordance with the procedure described in Example 18. EXAMPLE 33 (COMPOUND 80) 4-(3.4-Bis(cvclopentyloxy)phenyl)-3-(2-hvdroxy-2-propyl)isochromen-1-one

(64) (80)

The title compound was prepared from Compound (64) and cyclopentyl bromide in accordance with the procedure described in Example 18.

EXAMPLE 34

In accordance with the procedures described herein, the following compounds were made: 4-(4-Benzyloxy-3-methoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one,

COMPOUND (82); 4-(4-Hydroxy-3-methoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one,

COMPOUND (83);

4-(3-Benzyloxy-4-methoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one, COMPOUND (84);

4-(4-Cyclopentyloxy-3-methoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one,

COMPOUND (85);

4-(4-Butoxy-3-methoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one, COMPOUND (86); 2-(4-(3-(2-Hydroxy-2-propyl)-1-oxoisochromen-4-yl)-2-methoxyphenoxy)acetamide,

COMPOUND (87); 4-(4-sec-Butoxy-3-methoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one,

COMPOUND (88);

4-(3-Hydroxy-4-methoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one, COMPOUND (89);

4-(3,4-Dimethoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one, COMPOUND (90); 2-(5-(3-(2-Hydroxy-2-propyl)-1-oxoisochromen-4-yl)-2-methoxyphenoxy)acetamide,

COMPOUND (91); 4-(3-Butoxy-4-methoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one, COMPOUND

(92); 3-(2-Hydroxy-2-propyl)-4-(4-methoxy-3-(pyridin-4-ylmethoxy)phenyl)isochromen-1-one,

COMPOUND (93); 2-(4-(3-(2-Hydroxy-2-propyl)-1-oxoisochromen-4-yl)-2-methoxyphenoxy)aceticacid,

COMPOUND (94);

2-(5-(3-(2-Hydroxy-2-propyl)-1-oxoisochromen-4-yl)-2-methoxyphenoxy)aceticacid, COMPOUND (95);

4-(4-(2-benzyloxyethoxy)-3-methoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one,

COMPOUND (96); 4-(4-(2-Hydroxyethoxy)-3-methoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one,

COMPOUND (97); 4-(3-Benzyloxy-4-cyclopentyloxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one,

COMPOUND (98); 4-(4-Cyclopropylmethoxy-3-ethoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one,

COMPOUND (99);

2-(2-cyclopentyloxy-5-(3-(2-hydroxy-2-propyl)-1-oxoisochromen-4-yl)phenoxy)acet- amide, COMPOUND (100);

2-(2-Cyclopentyloxy-5-(3-(2-hydroxy-2-propyl)-1-oxoisochromen-4-yl)phenoxy)- propanamide, COMPOUND (101); 4-(4-Butoxy-3-ethoxyphenyl)-3-(2-hydroxy-2-propyl)isochromen-1-one, COMPOUND

(102); 4-(3-(2-Hydroxy-2-propyl)-1 -oxoisochromen-4-yl)-2-methoxyphenyl 4-methyl- benzenesulfonate, COMPOUND (103); 4-(4-Cyclopentyloxy-3-ethoxyphenyl)-3-(1-hydroxycyclopentyl)isochromen-1-one,

COMPOUND (107);

4-(4-Cyclopentyloxy-3-ethoxyphenyl)-3-(3-hydroxy-3-pentyl)isochromen-1-one, COMPOUND (109);

4-(4-Cyclopentyloxy-3-propoxyphenyl)-3-(3-hydroxy-3-pentyl)isochromen-1-one,

COMPOUND (110); 2-(2-Cyclopentyloxy-5-(3-(3-hydroxy-3-pentyl)-1-oxoisochromen-4-yl)phenoxy)acet- amide, COMPOUND (111 ); 4-(4-Cyclopentyloxy-3-ethoxyphenyl)isochromen-1 -one; 4-(4-Cyclopentyloxy-3-propoxyphenyl)-3-(1-hydroxycyclopentyl)isochromen-1-one,

COMPOUND (113); 2-(2-Cyclopentyloxy-5-(3-(1-hydroxycyclopentyl)-1-oxoisochromen-4-yl)phenoxy)acet- amide, COMPOUND (114); and 2-(2-Cyclopentyloxy-5-(1-oxoisochromen-4-yl)phenoxy)acetamide, COMPOUND (115).

BIOLOGICAL EXAMPLES

In vitro Inhibition of PDE4 and PDE7 phosphodiesterases

A. PDE4 and PDE7 Assays

The compounds were tested in vitro determining perturbations in cAMP consumption of PDE4 or PDE7, using a PDE assay based on the Phosphodiesterase [³H]CAMP Scintillation proximity assay (SPA), kit TRKQ 7090 from GE Healthcare. Compound solutions used in assay contained 1% DMSO.

PDE4:

Reactions in duplicate were initiated by the addition of 15 μl PDE4 (U937 lysate) to 10 μL of assay mix and 25 μL of test compounds in lsoplates (Wallac). The final assay mixture contained 50 mM Tris (pH 7.5), 8.3 mM MgCI₂, 1.7 mM EGTA and [³H]cAMP (0.025 μCi) (Amersham). The mixture was incubated at rt for 60 min, before quenching by addition of 25 μL SPA yttrium silicate beads/zinc sulphate. The plate was sealed, shaken for 1 min and then allowed to settle 20 min before determination of residual [³H]-CAMP using a Packard Topcount Scintillation counter.

PDE7: Reactions in duplicate were initiated by the addition of 15 μl PDE7 (recombinant human His-tagged PDE7 from SF9/Baculoviris expression) to 10 μL of assay mix and 25 μL of test compounds in lsoplates (Wallac). The final assay mixture contained 50 mM Tris (pH 7.5), 8.3 mM MgCI₂, 1.7 mM EGTA and [³H]CAMP (0.025 μCi) (Amersham). The mixture was incubated at rt for 60 min, before quenching by addition of 25 μL SPA yttrium silicate beads/zinc sulphate. The plate was sealed, shaken for 1 min and then allowed to settle 45 min before determination of residual [³H]-CAMP using a Packard Topcount Scintillation counter. B. Data evaluation and result

IC₅₀ values were determined from the concentration curves by non-linear regression analysis using Sigma Plot.

The compounds of invention demonstrated the ability to inhibit PDE7 phosphodiesterase activity and dual PDE4/7 inhibition.

The PDE4 and PDE7 inhibitory activity of representative compounds of formula (I) are shown in the following TABLE 1. The ratio of PDE4 to PDE7 activity demonstrates the selectivity of the compounds for the particular PDE. The compound numbers in the Table refer to the compounds disclosed herein as being prepared by the methods disclosed herein:

TABLE 1

COMPOUND PDE7 PDE4 PDE4 /PDE7

# IC₅₀ IC₅₀ ratio (uM) (uM)

43 4.11 2.33 0.57

44 3.12 3.83 1.23

45 8.45 7.67 0.91

47 7.06 2.5 0.35

82 2.49 4.33 1.74

83 4.93 11.87 2.41

84 8.02 5.36 0.67

85 1.84 7.66 4.16

86 2.43 7.96 3.28

87 3.42 10.93 3.2

88 1.84 10.27 5.58

89 6.23 13.83 2.22 COMPOUND PDE7 PDE4 PDE4 /PDE7

# IC₅₀ IC₅₀ ratio (uM) (uM)

90 4.32 42.54 9.85

91 3.86 46.95 12.16

92 7.54 7.13 0.95

93 8.46 9.48 1.12

94 3.3 12.82 3.88

95 21.89 >100 >4.57

96 2.52 4.27 1.69

97 3.5 6.8 1.94

49 >100 >100 -

50 4.13 3.82 0.92

51 6.95 2.33 0.34

52 5.59 4.29 0.77

53 3.19 2.14 0.67

60 6.42 26.8 4.17

61 4.25 8.8 2.07

62 3.4 6.93 2.04

63 3.51 5.23 1.49

64 3.6 5.74 1.59

67 2.7 16.29 6.03

68 9.11 17.52 1.92

69 2.55 11.46 4.49

99 2.1 5.81 2.77

71 5.45 7.99 1.47

65 2.71 9.02 3.33 COMPOUND PDE7 PDE4 PDE4 /PDE7

# IC₅₀ IC₅₀ ratio (uM) (uM)

66 4.4 3.52 0.8

72 2.81 4.26 1.52

100 1.2 12.92 10.77

77 3.43 13.65 3.98

78 1.43 2.89 2.02

73 1.79 10.03 5.6

74 5.77 9.65 1.67

75 2 4.62 2.31

80 1.97 2.76 1.4

79 3.47 3.21 0.93

Claims

1. A compound of formula (I):

wherein:

n represents 0, 1 , 2, 3 or 4; m represents 0, 1 , 2 or 3;

R¹ represents hydrogen, -C(OR⁴³XR⁴")^ -C(R^4b)₃ or -Si(R^6p)₃;

R² and R³ independently represent hydrogen, C_1-12 alkyl (optionally substituted by one or more substituents selected from A¹), aryl (optionally substituted by one or more substituents selected from A²), heteroaryl (optionally substituted by one or more substituents selected from A³), heterocycloalkyl (optionally substituted by one or more substituents selected from A⁴), -C(O)OR⁵³, -C(O)N(R^5b)R^5c, -S(O)₂R^5d or -C(O)R^5e;

each R^4a and R^4b independently represent, on each occasion when used herein, hydrogen or C_1-12 alkyl optionally substituted by one or more substituents selected from A⁵; or any two R^4b substituents when attached to a common carbon atom may be linked together to form a 3- to 12-membered ring, optionally containing one to three heteroatoms, one to three unsaturations and which ring is optionally substituted with one or more substituents selected from A⁶;

each R^x and R^y independently represents, on each occasion when used herein, A⁷ and/or C_1-12 alkyl optionally substituted by one or more substituents selected from A⁸;

A¹, A², A³, A⁴, A⁵, A⁶, A⁷ and A⁸ independently represent aryl (optionally substituted by one or more substituents selected from B¹), heteroaryl (optionally substituted by one or more substituents selected from B²) heterocycloalkyl (optionally substituted by one or more substituents selected from B³), halo, -CN₁ -NO₂, -C(O)OR⁶³, -C(O)N(R^7a)R^8a, -O-R^6b, -OC(O)R^6c, -N(R^7b)R^8b, -C(O)R^6d, -OS(O)₂R⁶⁸, -OC(O)N(R^7c)R^8c, -N(R^6f)C(O)OR⁶⁹, -N(R^6h)C(O)R⁶ⁱ, -N(R⁶ⁱ)S(O),R^6k, -S(O),OR^6m, -S(O)_pR⁶ⁿ, -S(O)_tN(R^7d)R^8d, -Si(R^6p)₃, -N(R^6q)C(O)N(R^7e)R^8e and/or -OC(O)OR^6r; and/or A¹, A⁴, A⁵, A⁶ and A⁸ may alternatively and independently represent =0;

t represents, on each occasion when used herein, 1 or 2;

p represents, on each occasion when used herein, O, 1 or 2;

D Γ\^6m , D Γ\⁶ⁿ , DΓ\⁶I , DΓ\^7a , DΓ\^7b , DΓΛ^7c , O Γ\^7d , DIΛ^7e ,

R^8a, R^8b, R^8c, R^8d and R^8e independently represent, on each occasion when used herein, hydrogen, C_1-12 alkyl (optionally substituted by one or more substituents selected from B⁴), aryl (optionally substituted by one or more substituents selected from B⁵), heteroaryl (optionally substituted by one or more substituents selected from B⁶) or heterocycloalkyl (optionally substituted by one or more substituents selected from B⁷); or any pair of R^5b and R^5c, R^7a and R^8a, R^7b and R^8b, R^7c and R^8c, R^7d and R^8d and R^7e and R^8e may be linked together to form, together with the nitrogen atom to which they are necessarily attached, a 3- to 12-membered ring, optionally containing a further one or two heteroatoms, one or two unsaturations, and which ring is optionally substituted by one or more susbtituents selected from B⁸;

R^5a, R^5d, R^6c, R^6e, R^6g, R^6k, R^6p and R^6r independently represent, on each occasion when used herein, C_1-12 alkyl (optionally substituted by one or more substituents selected from B⁴), aryl (optionally substituted by one or more substituents selected from B⁵), heteroaryl (optionally substituted by one or more substituents selected from B⁶) or heterocycloalkyl (optionally substituted by one or more substituents selected from B⁷);

B¹, B², B³, B⁴, B⁵, B⁶, B⁷ and B⁸ independently represent halo, C_1-6 alkyl (optionally substituted by one or more halo atoms), -OR^9a, -N(R^9b)R^10b, -NO₂, -CN, aryl (optionally substituted by one or more substituents selected from E¹), heteroaryl (optionally substituted by one or more substituents selected from E²) and/or heterocycloalkyl (optionally substituted by one or more substituents selected from E³); and/or B³, B⁴, B⁷ and B⁸ may alternatively and independently represent =0;

R^9a, R^9b and R^1Ob independently represent hydrogen or C_1-6 alkyl optionally substituted by one or substituents selected from E⁴;

E¹, E², E³ and E⁴ independently represent halo, -CN, -OH, -OC_1-6 alkyl (optionally substituted by one or more fluoro atoms) and/or C_1-6 alkyl (optionally substituted by one or more halo atoms),

or a pharmaceutically acceptable salt thereof.

2. A compound as claimed in Claim 1 , wherein R¹ represents hydrogen or -C(OR⁴³XR⁴"),.

3. A compound as claimed in Claim 1 or Claim 2, wherein R² represents hydrogen or C_1-12 alkyl (optionally substituted by one or more substituents selected from A¹).

4. A compound as claimed in any one of the preceding claims, wherein R^4a represents C_1-3 alkyl or hydrogen.

5. A compound as claimed in any one of the preceding claims, wherein each R^4b independently represents hydrogen or Ci_-6 alkyl optionally substituted by one or more substituents selected from A⁵; or two R^4b groups when attached to a common carbon atom are linked together to form a 3- to 8-membered ring, optionally containing one heteroatom, which ring may be substituted by one or more A⁶ substituents.

6. A compound as claimed in any one of the preceding claims, wherein R³ represents hydrogen, C_1-^ alkyl (optionally substituted by one or more substituents selected from A¹) or -S(O)₂R^5d.

7. A compound as claimed in any one of the preceding claims, wherein A¹ to A⁸ independently represent aryl (optionally substituted by one or more B¹ substituents), heteroaryl (optionally substituted by one or more substituents selected from B²), heterocycloalkyl (optionally substituted by one or more substituents selected from B³), halo, -CN, -NO₂, -C(O)OR^6a, -O-R^6b, -N(R^7b)R^8b or -C(O)N(R^7a)R^8a.

8. A compound as claimed in any one of the preceding claims, wherein R^5b, p5c p6a p6b p6d p6f p6h p6i p6j p6m p6n p6p p6q p7a p7b . p7c p7d p7e p8a R^8b, R^8c, R^8d and R^8e independently represent hydrogen, C_1-6 alkyl (optionally substituted by one or more substituents selected from B⁴) or aryl (optionally substituted by one or more substituents selected from B⁵); or any pair of R^5b and R⁵⁰, R^7a and R^8a, R^7b and R^8b, R^7c and R^8c, R^7d and R^8d and R^7e and R^8e may be linked together to form, together with the nitrogen atom to which they are necessarily attached, a 3- to 8- membered ring, optionally containing a further heteroatom, which ring is optionally substituted by one or two substituent(s) selected from B⁸.

9. A compound as claimed in any one of the preceding claims, wherein R^5a, R^5d, R^6c, R^6e, R⁶⁹, R^6k and R^6r independently represent Ci_-6 alkyl (optionally substituted by one or more substituents selected from B⁴) or aryl (optionally substituted by one or more substituents selected from B⁵).

10. A compound as claimed in any one of the preceding claims, wherein B¹, B², B³, B⁴, B⁵, B⁶, B⁷ and B⁸ independently represent halo, C_1-6 alkyl (optionally substituted by one or more halo atoms) and/or aryl (optionally substituted by one or more substituents selected from E¹); and/or B³, B⁴, B⁷ and B⁸ may alternatively and independently represent =0.

11. A compound as claimed in any one of the preceding claims, wherein R^9a, R^9b and R^1Ob independently represent hydrogen or C_1-3 (e.g. Ci_-2) alkyl optionally substituted by one or substituents selected from E⁴.

12. A compound as claimed in any one of the preceding claims, wherein E¹, E², E³ and E⁴ independently represent halo or Ci_-2 alkyl.

13. A compound of formula I as defined in any one of Claims 1 to 12, or a pharmaceutically acceptable salt thereof, for use as a pharmaceutical.

14. A pharmaceutical formulation including a compound of formula I, as defined in any one of Claims 1 to 12, or a pharmaceutically acceptable salt thereof, in admixture with a pharmaceutically acceptable adjuvant, diluent, carrier or excipient.

15. A compound, as defined in any one of Claims 1 to 12, or a pharmaceutically acceptable salt thereof, for use in the treatment of: i) an inflammatory disorder; ii) a disorder in which the modulation of intracellular cyclic adenosine 5'-monophosphate levels within a mammal is desired and/or required, which disorder may be an inflammatory disorder; iii) a disorder associated with pathological conditions that are modulated by inhibiting enzymes associated with secondary cellular messengers; iv) transplant rejection in a mammal; v) uncontrolled cellular proliferation; and/or vi) a disorder associated with the central nervous system.

16. Use of a compound of formula I, as defined in any one of Claims 1 to 12, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of a disorder as defined by any of i) to vi) in Claim 15.

17. A compound as claimed in Claim 15 or a use as claimed in Claim 16, wherein the compound is an inhibitor of PDE4, PDE7 or a dual inhibitor of PDE4 and PDE7.

18. A compound as claimed in Claim 15 or a use as claimed in Claim 16 (or a compound or use as claimed in Claim 17), wherein the disorder is inflammation, a proliferative disorder or a disease or pathological condition of the central nervous system.

19. A compound or use as claimed in Claim 18, wherein the disorder is ankylosing spondylitis, arthritis, asthma, chronic obstructive pulmonary disease, chronic bronchitis, respiratory distress syndrome, rhinitis, allergic rhinitis, Crohn's disease, nephritis, eczema, atopic dermatitis, urticaria, conjunctivitis, ulcerative colitis, rheumatoid arthritis, osteoarthritis, eosinophilic gastrointestinal disorders, vascular disease, diabetes mellitus, fibromyalgia syndrome, gout, inflammations of the brain, emphysema, inflammatory bowel disease, irritable bowel syndrome, ischemia-reperfusion injury juvenile erythematosus pulmonary sarcoidosis, Kawasaki disease, osteoarthritis, pelvic inflammatory disease, psoriatic arthritis, rheumatoid arthritis, psoriasis, tissue/organ transplant, scleroderma, spondyloarthropathies, systemic lupus erythematosus, pulmonary sarcoidosis, ulcerative colitis, a vial infection, a bacterial infection, cancer, leukemia, a solid tumor, cognitive function, Alzheimer's disease, a learning and memory disorder, cerebrovascular disease, depression, schizophrenia, Parkinson's disease and/or multiple sclerosis.

20. A compound or use as claimed in Claim 18, wherein the disorder is a cutaneous disease.

21. A compound or use as claimed in Claim 20, wherein the disorder is psoriasis, eczema, dermatitis and/or urticaria.

22. A method of treatment of a disorder as defined by i) to vi) in Claim 15, which method comprises administration of a therapeutically effective amount of a compound of formula I as defined in any one of Claims 1 to 12, or a pharmaceutically-acceptable salt thereof, to a patient suffering from, or susceptible to, such a condition.

23. A method as claimed in Claim 22, wherein the disorder is one in which the inhibition (or modulation) of PDE4, PDE7 or PDE4 and PDE7 is desired and/or required.

24. A combination product comprising: (A) a compound of formula I as defined in any one of Claims 1 to 12, or a pharmaceutically-acceptable salt thereof; and

(B) another therapeutic agent that is useful in the treatment of a disorder as defined by i), ii), iii), iv), v) or vi) in Claim 15, wherein each of components (A) and (B) is formulated in admixture with a pharmaceutically-acceptable adjuvant, diluent, carrier or excipient.

25. A combination product as claimed in Claim 24 which comprises a pharmaceutical formulation including a compound of formula I as defined in any one of Claims 1 to 12, or a pharmaceutically-acceptable salt thereof, another therapeutic agent that is useful in the treatment of a disorder as defined by i), ii), iii), iv), v) or vi) in Claim 15, and a pharmaceutically-acceptable adjuvant, diluent, carrier or excipient.

26. A combination product as claimed in Claim 24 which comprises a kit of parts comprising components:

(a) a pharmaceutical formulation including a compound of formula I as defined in any one of Claims 1 to 12, or a pharmaceutically-acceptable salt thereof, in admixture with a pharmaceutically-acceptable adjuvant, diluent, carrier or excipient; and (b) a pharmaceutical formulation including another therapeutic agent that is useful in the treatment of a disorder as defined by i), ii), iii), iv), v) or vi) in Claim 15 in admixture with a pharmaceutically-acceptable adjuvant, diluent, carrier or excipient, which components (a) and (b) are each provided in a form that is suitable for administration in conjunction with the other.

27. A combination product as claimed in any one of Claims 24 to 26, wherein the other therapeutic agent is useful in the treatment of a disorder in which the inhibition (or modulation) of PDE4, PDE7 or PDE4 and PDE7 is desired and/or required.

28. A process for the preparation of a compound of the formula I as defined in Claim 1 which process comprises:

(i) reaction of a compound of formula II,

wherein R¹, R², R³, R^y and m are as defined in Claim 1 , with a compound of formula III,

wherein L¹ represents a suitable leaving group, R^s1 represents C_1-6 alkyl optionally substituted by one or more halo atoms, and R^x and n are as defined in Claim 1 ; (ii) for compounds of formula I in which R¹ represents hydrogen, de-silylation of a compound corresponding to a compound of formula I but in which R¹ represents -Si(R^t1)₃, and each R^t1 independently represents a Ci_-6 alkyl group; (iii) for compounds of formula I in which R² and/or R³ are other than hydrogen, reaction of a corresponding compound of formula I in which R² and/or R³ (as appropriate) do represent hydrogen, with a compound of formula IV,

_R2/3_|_x IV wherein L^x represents a suitable leaving group, and R²⁷³ represents either R² or R³, provided that they do not represent hydrogen;

(iv) for compounds of formula I in which R² and/or R³ are hydrogen, reaction of a corresponding compound of formula I in which R² and/or R³ (as appropriate) do not represent hydrogen with a strong acid, a thiolate anion or by hydrogenolysis.

29. A process for the preparation of a pharmaceutical formulation as defined in Claim 14, which process comprises bringing into association a compound of formula I₁ as defined in any one of Claims 1 to 12, or a pharmaceutically acceptable salt thereof with a pharmaceutically-acceptable adjuvant, diluent, carrier or excipient.

30. A process for the preparation of a combination product as defined in any one of Claims 24 to 27, which process comprises bringing into association a compound of formula I, as defined in any one of Claims 1 to 12, or a pharmaceutically acceptable salt thereof with the other therapeutic agent that is useful in the treatment of a disorder as defined by i), ii), iii), iv), v) or vi) in Claim 15, or, useful in the treatment of a disorder in which the inhibition (or modulation) of PDE4, PDE7 or PDE4 and PDE7 is desired and/or required, and at least one pharmaceutically-acceptable adjuvant, diluent, carrier or excipient.