WO2009153536A1 - 1-aza-bicyclo [2.2.2] octane derivatives useful as muscarinic receptor antagonists - Google Patents

Abstract

The invention provides named compounds of formula (I), pharmaceutical compositions containing them, a process for preparing the pharmaceutical compositions and their use in therapy.

Description

1 -AZA-BICYCLO [2 . 2 . 2 ] OCTANE DERIVATIVES USEFUL AS MUSCARINIC RECEPTOR

ANTAGONISTS

The present invention relates to N-linked-heterocycle-substituted alkyl esters of polycyclic amino alcohols, a process for their preparation, pharmaceutical compositions containing them, a process for preparing pharmaceutical compositions, their use in therapy and intermediates of use in their preparation.

Muscarinic receptors are a G-protein coupled receptor (GPCR) family having five family members Mj, M₂, M₃, M₄ and M₅. Of the five muscarinic subtypes, three (Mi, M₂ and M₃) are known to exert physiological effects on human lung tissue.

Parasympathetic nerves are the main pathway for reflex bronchoconstriction in human airways and mediate airway tone by releasing acetylcholine onto muscarinic receptors. Airway tone is increased in patients with respiratory disorders such as asthma and chronic obstructive pulmonary disease (COPD), and for this reason muscarinic receptor antagonists have been developed for use in treating airway diseases. Muscarinic receptor antagonsists, often called anticholinergics in clinical practice, have gained widespread acceptance as a first-line therapy for individuals with COPD, and their use has been extensivley reviewed in the literature (e.g. Lee et al, Current Opinion in Pharmacology 2001,1, 223-229).

When used to treat respiratory disorders, muscarinic receptor antagonists are typically administered by inhalation. However, when administered by inhalation a significant proportion of the muscarinic receptor antagonist is often absorbed into the systemic circulation resulting in reported side effects such as dry mouth. Additionally, the majority of muscarinic antagonists have a relatively short duration of action requiring that they be administered several times a day. Such a multiple-daily dosing regime is not only inconvenient to the patient but also creates a significant risk of inadequate treatment due to patient non-compliance associated with the frequent repeat dosing schedule.

There therefore remains a need for novel compounds that are capable of blocking muscarinic receptors. In particular, a need exists for new muscarinic antagonists that have high potency and reduced systemic side effects when administered by inhalation. Moreover, a need exists for new muscarinic antagonists that exhibit a long duration of action when dosed by inhalation, and which are amenable to either once or twice daily dosing.

Our co-pending application PCT/GB 2007/004817 relates to compounds of formula (I):

wherein

R¹ represents C_1-6 alkyl;

R² represents phenyl or a 5 to 6 membered heteroaryl ring, each of which may be optionally substituted by one or more substituents independently selected from halogen, cyano, nitro, SH, S(O)_0-2R¹⁰, NR¹¹R¹², S(O)₂NR¹³R¹⁴, C(O)NR¹⁵R¹⁶, C(O)₂R¹⁷, NR¹⁸S(O)₂R¹⁹, NR²⁰C(O)R²¹, NR²²C(O)₂R²³, NR²⁴C(O)NR²⁵R²⁶, OR²⁷ and C_1-6 alkyl which C)_-6 alkyl may be optionally substituted by one or more substituents independently selected from halogen, hydroxyl, Ci_-6 alkoxy, NH₂, NH(Ci_-6 alkyl) and N(Cj_-6 alkyl)₂; R³ and R⁴ together with the nitrogen atom to which they are both attached form a 4 to 8 membered aliphatic heterocyclic ring which may optionally contain a further heteroatom selected from oxygen, sulphur and nitrogen, and which heterocyclic ring may be optionally substituted by one or more substituents independently selected from halogen, hydroxyl, Q- ₆alkoxy and C]_-6 alkyl, which C]_-6 alkyl may be optionally substituted by one or more substituents independently selected from halogen and hydroxyl; R⁵ represents a group of formula (II) or (III);

wherein

Y is -CH₂-, -CH₂CH₂- or -CH₂CH₂CH₂- and the substitution on the ring in group (II) is in the 3 or 4 positions; a is 1 or 2; b is 1 or 2;

Z is -CH₂-;

R⁶ represents a group of formula (IV)

wherein w is 0 or 1 ;

R⁷ represents Ci_-4 alkylene optionally substituted by one or more substituents independently selected from halogen, hydroxyl, Ci_-6 alkoxy, NH₂, NH(Ci_-6 alkyl) and

N(C_-6 alkyl)₂; when w is 0, y is 0; when w is 1, y is 0 or 1;

Q represents O, C(O), S(O)_0-2, NR⁹, -CONR⁹-, -SO₂NR⁹-, -NR⁹CO-, -NR⁹SO₂-, -OC(O)-, -

C(O)O-, -HC=CH- or ethynylene; R⁸ represents a cyclic group Cyc¹ or a Ci_-4 alkyl group which Ci_-4 alkyl group may be optionally substituted by one or more substituents independently selected from halogen, hydroxyl, Ci_-4 alkoxy, NH₂, NH(Ci_-4 alkyl), N(C_!-4 alkyl)₂, a cyclic group Cyc² and OCyc²; and R⁸ may additionally represent hydrogen when Q represents O, NR⁹, -CONR⁹-, -

SO₂NR⁹-, -C(O)O-, -HC=CH- or ethynylene; Cyc¹ and Cyc² each independently represent aryl, heteroaryl, a 3 to 8 membered aliphatic carbocyclic ring or a 4 to 8 membered aliphatic heterocyclic ring, each of which may be optionally substituted by one or more substituents independently selected from halogen, cyano, nitro, SH, S(O)_0-2R¹⁰, NR¹¹R¹², S(O)₂NR¹³R¹⁴, C(O)NR¹⁵R¹⁶, C(O)₂R¹⁷,

NR¹⁸S(O)₂R¹⁹, NR²⁰C(O)R²¹, NR²²C(O)₂R²³, NR²⁴C(O)NR²⁵R²⁶, OR²⁷, phenyl, heteroaryl and Ci_-6 alkyl which phenyl, heteroaryl or C_)-6 alkyl may be optionally substituted by one or more substituents independently selected from halogen, hydroxyl, cyano, Ci_-6 alkoxy,

NH₂, NH(Ci_-6 alkyl) and N(C_1-6 alkyl)₂;

R⁹ represents hydrogen or Ci_-6 alkyl;

R¹⁰ and R¹⁹ each independently represent Ci_-6 alkyl, which Ci_-6 alkyl may be optionally substituted by one or more substituents independently selected from halogen, hydroxyl, C-

6 alkoxy, NH₂, NH(C_-6 alkyl) and N(C_-6 alkyl)₂; and R", R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R²⁰, R²¹, R²², R²³, R²⁴, R²⁵, R²⁶ and R²⁷ each independently represent hydrogen or C_1-6 alkyl, which Ci_-6 alkyl may be optionally substituted by one or more substituents independently selected from halogen, hydroxyl, Q- ₆ alkoxy, NH₂, NH(C_1-6 alkyl) and N(C_1-6 alkyl)₂; or any of R¹ ' and R¹², R¹³ and R¹⁴, R¹⁵and R¹⁶ or R²⁵ and R²⁶, together with the nitrogen atom to which they are both attached, may form a 4 to 8 membered aliphatic heterocyclic ring, which heterocyclic ring may be optionally substituted by one or more substituents independently selected from halogen, hydroxyl and C]_-6 alkyl, which C_1-6 alkyl may be optionally substituted by one or more substituents independently selected from halogen and hydroxyl; and X represents a pharmaceutically acceptable anion of a mono or polyvalent acid.

Summary of the Invention

The present invention provides compounds falling within the scope of, but not specifically disclosed in, our co-pending application PCT/GB2007/004817 referred to above.

Thus, in one aspect, the present invention provides a compound selected from the group consisting of:

CR)- 1 -[2-(2-Fluoro-phenyl)-ethyl]-3-((S)-2-phenyl-2-piperidin- 1-yl-propionyloxy)- 1 - azonia-bicyclo[2.2.2]octane X;

(/?)-l-[2-(2-Fluoro-ρhenyl)-ethyl]-3-((6')-2-ρiρeridin-l-yl-2-thiophen-2-yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(i?)-l-[2-(2-Fluoro-phenyl)-ethyl]-3-((i?)-2-piperidin-l-yl-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X; (R)- l-(6-Methyl-benzooxazol-2-ylmethyl)-3-((S)-2-phenyl-2-piρeri din- 1-yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(/?)-l-(3-Benzooxazol-2-yl-propyl)-3-((5')-2-phenyl-2-piperidin-l-yl-ρropionyloxy)-l- azonia-bicyclo[2.2.2]octane X;

(R)- 1 -(5-Chloro-benzooxazol-2-ylmethyl)-3-((,S)-2-phenyl-2-piperidin- 1 -yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(i?)-l-(6-Chloro-benzooxazol-2-ylmethyl)-3-((₁S')-2-phenyl-2-piperidin-l-yl- propionyloxy)- 1 -azonia-bicyclo[2.2.2]octane X; (R)- 1 -Benzooxazol-2-ylmethyl-3-((S)-2-phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 - azonia-bicyclo[2.2.2]octane X;

(R)-l-(5-Methyl-benzooxazol-2-ylmethyl)-3-((5)-2-phenyl-2-piperidin-l-yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X; (i?)-3-((5)-2-Phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 -( 1 -phenyl- lH-pyrazol-4- ylmethyl)-l-azonia-bicyclo[2.2.2]octane X;

(R)-3-((5')-2-Phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 -( 1 -phenyl- IH-[1 ,2,3]triazol-4- ylmethyl)- l-azonia-bicyclo[2.2.2]octane X;

(R)- 1 -Isopropyl-3-((5')-2-phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 -azonia- bicyclo[2.2.2]octane X;

(i?)-l-(5-Phenyl-[l,2,4]oxadiazol-3-ylmethyl)-3-((5')-2-piperidin-l-yl-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(R)- l-(5-Phenyl-[ 1 ,2,4]oxadiazol-3-ylmethyl)-3-((i?)-2-ρiρeridin- l-yl-2-thiophen-2-yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X; (R)- 1 -(3-Phenyl-isoxazol-5-ylmethyl)-3-((5')-2-piperidin- 1 -yl-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(i?)-l-(3-Phenyl-isoxazol-5-ylmethyl)-3-((R)-2-piperidin-l-yl-2-thiophen-2-yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(R)- l-[3-(4-Methoxy-phenyl)-[ 1 ,2,4]oxadiazol-5-ylmethyl]-3-((.S)-2-piperidin- 1 -yl-2- thiophen-2-yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(i?)-l-[3-(4-Methoxy-phenyl)-[l,2,4]oxadiazol-5-ylmethyl]-3-((i?)-2-piperidin-l-yl-2- thiophen-2-yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(/?)-l-[3-(5-Methyl-ρyridin-3-yl)-propyl]-3-((6')-2-pipeπdin-l-yl-2-thiophen-2-yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X; (/?)-l-[3-(5-Methyl-pyridin-3-yl)-propyl]-3-((/?)-2-piperidin-l-yl-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(R)-l-(5-Phenyl-isoxazol-3-ylmethyl)-3-((5)-2-piρeridin-l-yl-2-thioρhen-2-yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(R)- 1 -(5-Phenyl-isoxazol-3-ylmethyl)-3-((/?)-2-piperidin- 1 -yl-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(R)- 1 - { 2-[4-(2-Ηydroxy-ethyl)-phenyl]-ethyl } -3-((>S)-2-phenyl-2-piperidin- 1 -yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X; (/?)-l-[2-(lH-Indol-3-yl)-ethyl]-3-((S)-2-phenyl-2-piperidin-l-yl-proρionyloxy)-l- azonia-bicyclo[2.2.2]octane X;

(R)- 1 -(5 -Phenyl-oxazol-2-ylmethyl)-3-((5)-2-phenyl-2-piperidin- 1 -yl-propionyloxy)- l-azonia-bicyclo[2.2.2]octane X; (i?)-l-(5-Phenyl-isoxazol-3-ylmethyl)-3-((5')-2-ρhenyl-2-piperidin-l-yl-propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(jR)-l-(3-Phenyl-isoxazol-5-ylmethyl)-3-((S)-2-phenyl-2-piperidin-l-yl-propionyloxy)- 1 -azonia-bicyclo[2.2.2]octane X;

(Ry 1 -(3-Phenyl-[ 1 ,2,4]oxadiazol-5-ylmethyl)-3-((5)-2-phenyl-2-piperidin-l -yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(i?)-l-(5-Phenyl-[l,2,4]oxadiazol-3-ylmethyl)-3-((S)-2-phenyl-2-piperidin-l-yl- propionyloxy)~l-azonia-bicyclo[2.2.2]octane X;

(i?)-l-(2-Pb.enyl-oxazol-4-ylmethyl)-3-((₁S)-2-phenyl-2-piperidin-l-yl-propionyloxy)- 1 -azonia-bicyclo[2.2.2]octane X; (i?)-l-[3-(3-Fluoro-phenyl)-[l,2,4]oxadiazol-5-ylmethyl]-3-((_ιS)-2-ρhenyl-2-ρiperidin-

1 -yl-propionyloxy)- 1 -azonia-bicyclo [2.2.2] octane X;

(i?)-l-(2-Phenyl-oxazol-5-ylmethyl)-3-((S)'2-phenyl-2-piperidin-l-yl-propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(2?)-3-((5)-2-Phenyl-2-piperidin-l-yl-propionyloxy)-l-(3-pyridin-4-yl- [1 ^^loxadiazol-S-ylmethylH-azonia-bicycloj^^joctane X;

(Ry 1- { 2~[2-(4-Chloro-phenyl)-thiazol~4-yl]-ethyl } -3-((5')-2-phenyl-2-ρiperidin- 1 -yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(i?)-l-(3-Benzo[b]thioρhen-2-yl-propyl)-3-((S)-2-ρhenyl-2-piρeridin-l-yl- propionyloxy) - 1 -azonia-bicyclo [2.2.2]octane X; (/?)-3-((5)-2-Phenyl-2-piperidin-l-yl-propionyloxy)-l-(3-quinoxalin-6-yl-propyl)-l- azonia-bicyclo[2.2.2]octane X;

(i?)-l-[3-(6-Methyl-ρyridin-3-yl)-ρroρyl]-3-((S)-2-phenyl-2-ρiperidm-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(i?)-3-((5)-2-Phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 -(3-pyridin-4-yl-propyl)~ 1 - azonia-bicyclo[2.2.2]octane X; and

(i?)-l-[3-(2-Methyl-pyridin-4-yl)-propyl]-3-((5)-2-phenyl-2-ρiρeridin-l-yl- propionyloxy)- 1 -azonia-bicyclo[2.2.2]octane X; wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid.

The compounds of formula (I), referred to above, and those of the present invention comprise an anion X associated with the positive charge on the quaternary nitrogen atom. The anion X may be any pharmaceutically acceptable anion of a mono or polyvalent (e.g. bivalent) acid. In an embodiment of the invention X may be an anion of a mineral acid, for example chloride, bromide, iodide, sulfate, nitrate or phosphate; or an anion of a suitable organic acid, for example acetate, maleate, fumarate, citrate, oxalate, succinate, tartrate, methanesulphonate, p-toluenesulphonate, benzenesulphonate, napadisylate (naphthalene- 1,5-disulfonate) (e.g. a heminapadisylate), 2,5-dichlorobenzenesulphonate, 1- hydroxynaphthalene-2-sulphonate or xinafoate (l-hydroxy-2-naphthoate).

It will be understood that certain compounds of the present invention may exist in solvated, for example hydrated, as well as unsolvated forms. It is to be understood that the present invention encompasses all such solvated forms.

The compounds of the present invention are capable of existing in stereoisomeric forms. It will be understood that the invention encompasses all geometric and optical isomers of the compounds of the present invention and mixtures thereof including racemates. Tautomers and mixtures thereof also form an aspect of the present invention.

In one embodiment, the present invention comprises compounds in optically pure form.

In the context of the present specification, the term optically pure is defined in terms of enantiomeric excess (e.e.), which is calculated from the ratio of the difference between the amounts of the respective enantiomers present and the sum of these amounts, expressed as a percentage. To illustrate, a preparation containing 95% of one enantiomer and 5% of another enantiomer has an enantiomeric excess (e.e.) of 90% [i.e. (95-5)/(95+5) x 100]. Optically pure compounds according to the present invention have an e.e. of at least 90%. In an embodiment of the invention, optically pure compounds have an e.e. of at least 95%. In a further embodiment of the invention, optically pure compounds have an e.e. of at least 98%. Where the compound has a diastereoisomer, optically pure compounds have an e.e.of at least 90% and a diastereomeric excess (d.e.) of at least 90 % [diastereomeric excess being defined by analogy to enantiomeric excess]. In an embodiment of the invention, optically pure compounds have an e.e. of at least 95% and a d.e.of at least 95%. In a further embodiment of the invention, optically pure compounds have an e.e. of at least 98% and a d.e. of at least 98%.

In one embodiment, the present invention provides the compound (i?)-l-[2-(2-Fluoro- phenyl)-ethyl]-3-((S)-2-phenyl-2-piperidin-l-yl-propionyloxy)-l-azonia- bicyclo[2.2.2]octane X, wherein X is a pharmaceutically acceptable anion of a mono or polyvalent acid. In one aspect of this embodiment X is bromide.

In one embodiment, the present invention provides the compound (i?)-l-(6-Methyl- benzooxazol-2-ylmethyl)-3-((S)-2-phenyl-2-piperidin-l-yl-propionyloxy)-l-azonia- bicyclo[2.2.2]octane X, wherein X is a pharmaceutically acceptable anion of a mono or polyvalent acid. In one aspect of this embodiment X is chloride.

In one embodiment, the present invention provides the compound (/?)-l-(5-Phenyl- isoxazol-3-ylmethyl)-3-((5)-2-phenyl-2-piperidin-l-yl-propionyloxy)-l-azonia- bicyclo[2.2.2]octane X,_wherein X is a pharmaceutically acceptable anion of a mono or polyvalent acid. In one aspect of this embodiment X is chloride.

In one embodiment, the present invention provides the compound (/?)-l-(3- Benzo[b]thiophen-2-yl-propyl)-3-((5)-2-phenyl-2-piperidin-l-yl-propionyloxy)-l-azonia- bicyclo[2.2.2]octane X, wherein X is a pharmaceutically acceptable anion of a mono or polyvalent acid. In one aspect of this embodiment X is bromide.

In one embodiment, the present invention provides the compound (/?)-3-((ιS)-2-Phenyl-2- piperidin- 1 -yl-propionyloxy)- 1 -(3-pyridin-4-yl-propyl)- 1 -azonia-bicyclo[2.2.2]octane, wherein X is a pharmaceutically acceptable anion of a mono or polyvalent acid. In one aspect of this embodiment X is bromide. In one embodiment, the present invention provides the compound (i?)-l-[3~(2-Methyl- pyridin-4-yl)-propyl]-3-((S)-2-phenyl-2-piperidin-l-yl-propionyloxy)-l-azonia- bicyclo[2.2.2]octane X, wherein X is a pharmaceutically acceptable anion of a mono or polyvalent acid. In one aspect of this embodiment X is bromide.

The compounds of the invention have activity as pharmaceuticals, in particular as anticholinergic agents including muscarinic receptor (Ml, M2, and M3) antagonists, in particular M3 antagonists. Diseases and conditions which may be treated with the compounds include: 1. respiratory tract: obstructive diseases of the airways including: asthma, including bronchial, allergic, intrinsic, extrinsic, exercise-induced, drug-induced (including aspirin and NSAID-induced) and dust-induced asthma, both intermittent and persistent and of all severities, and other causes of airway hyper-responsiveness; chronic obstructive pulmonary disease (COPD); bronchitis, including infectious and eosinophilic bronchitis; emphysema; bronchiectasis; cystic fibrosis; sarcoidosis; farmer's lung and related diseases; hypersensitivity pneumonitis; lung fibrosis, including cryptogenic fibrosing alveolitis, idiopathic interstitial pneumonias, fibrosis complicating anti-neoplastic therapy and chronic infection, including tuberculosis and aspergillosis and other fungal infections; complications of lung transplantation; vasculitic and thrombotic disorders of the lung vasculature, and pulmonary hypertension; antitussive activity including treatment of chronic cough associated with inflammatory and secretory conditions of the airways, and iatrogenic cough; acute and chronic rhinitis including rhinitis medicamentosa, and vasomotor rhinitis; perennial and seasonal allergic rhinitis including rhinitis nervosa (hay fever); nasal polyposis; acute viral infection including the common cold, and infection due to respiratory syncytial virus, influenza, coronavirus (including SARS) and adenovirus; 2. bone and joints: arthritides associated with or including osteoarthritis/osteoarthrosis, both primary and secondary to, for example, congenital hip dysplasia; cervical and lumbar spondylitis, and low back and neck pain; rheumatoid arthritis and Still's disease; seronegative spondyloarthropathies including ankylosing spondylitis, psoriatic arthritis, reactive arthritis and undifferentiated spondarthropathy; septic arthritis and other infection- related arthopathies and bone disorders such as tuberculosis, including Potts' disease and Poncet's syndrome; acute and chronic crystal-induced synovitis including urate gout, calcium pyrophosphate deposition disease, and calcium apatite related tendon, bursal and synovial inflammation; Behcet's disease; primary and secondary Sjogren's syndrome; systemic sclerosis and limited scleroderma; systemic lupus erythematosus, mixed connective tissue disease, and undifferentiated connective tissue disease; inflammatory myopathies including dermatomyositits and polymyositis; polymalgia rheumatica; juvenile arthritis including idiopathic inflammatory arthritides of whatever joint distribution and associated syndromes, and rheumatic fever and its systemic complications; vasculitides including giant cell arteritis, Takayasu's arteritis, Churg-Strauss syndrome, polyarteritis nodosa, microscopic polyarteritis, and vasculitides associated with viral infection, hypersensitivity reactions, cryoglobulins, and paraproteins; low back pain; Familial Mediterranean fever, Muckle- Wells syndrome, and Familial Hibernian Fever, Kikuchi disease; drug-induced arthalgias, tendonititides, and myopathies;

3. pain and connective tissue remodelling of musculoskeletal disorders due to injury [for example sports injury] or disease: arthitides (for example rheumatoid arthritis, osteoarthritis, gout or crystal arthropathy), other joint disease (such as intervertebral disc degeneration or temporomandibular joint degeneration), bone remodelling disease (such as osteoporosis, Paget's disease or osteonecrosis), polychondritits, scleroderma, mixed connective tissue disorder, spondyloarthropathies or periodontal disease (such as periodontitis); 4. skin: psoriasis, atopic dermatitis, contact dermatitis or other eczematous dermatoses, and delayed-type hypersensitivity reactions; phyto- and photodermatitis; seborrhoeic dermatitis, dermatitis herpetiformis, lichen planus, lichen sclerosus et atrophica, pyoderma gangrenosum, skin sarcoid, discoid lupus erythematosus, pemphigus, pemphigoid, epidermolysis bullosa, urticaria, angioedema, vasculitides, toxic erythemas, cutaneous eosinophilias, alopecia areata, male-pattern baldness, Sweet's syndrome, Weber-Christian syndrome, erythema multiforme; cellulitis, both infective and non-infective; panniculitis;cutaneous lymphomas, non-melanoma skin cancer and other dysplastic lesions; drug-induced disorders including fixed drug eruptions; 5. eyes: blepharitis; conjunctivitis, including perennial and vernal allergic conjunctivitis; iritis; anterior and posterior uveitis; choroiditis; autoimmune; degenerative or inflammatory disorders affecting the retina; ophthalmitis including sympathetic ophthalmitis; sarcoidosis; infections including viral , fungal, and bacterial; 02072

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6. gastrointestinal tract: glossitis, gingivitis, periodontitis; oesophagitis, including reflux; eosinophilic gastro-enteritis, mastocytosis, Crohn's disease, colitis including ulcerative colitis, proctitis, pruritis ani; coeliac disease, irritable bowel syndrome, and food-related allergies which may have effects remote from the gut (for example migraine, rhinitis or

5 eczema);

7. abdominal: hepatitis, including autoimmune, alcoholic and viral; fibrosis and cirrhosis of the liver; cholecystitis; pancreatitis, both acute and chronic;

8. genitourinary: nephritis including interstitial and glomerulonephritis; nephrotic syndrome; cystitis including acute and chronic (interstitial) cystitis and Hunner's ulcer;o acute and chronic urethritis, prostatitis, epididymitis, oophoritis and salpingitis; vulvovaginitis; Peyronie's disease; erectile dysfunction (both male and female);

9. allograft rejection: acute and chronic following, for example, transplantation of kidney, heart, liver, lung, bone marrow, skin or cornea or following blood transfusion; or chronic graft versus host disease; s 10. CNS: Alzheimer's disease and other dementing disorders including CJD and nvCJD; amyloidosis; multiple sclerosis and other demyelinating syndromes; cerebral atherosclerosis and vasculitis; temporal arteritis; myasthenia gravis; acute and chronic pain (acute, intermittent or persistent, whether of central or peripheral origin) including visceral pain, headache, migraine, trigeminal neuralgia, atypical facial pain, joint and bone pain,o pain arising from cancer and tumor invasion, neuropathic pain syndromes including diabetic, post-herpetic, and HIV-associated neuropathies; neurosarcoidosis; central and peripheral nervous system complications of malignant, infectious or autoimmune processes;

11. other auto-immune and allergic disorders including Hashimoto's thyroiditis, Graves'5 disease, Addison's disease, diabetes mellitus, idiopathic thrombocytopaenic purpura, eosinophilic fasciitis, hyper-IgE syndrome, antiphospholipid syndrome;

12. other disorders with an inflammatory or immunological component; including acquired immune deficiency syndrome (AIDS), leprosy, Sezary syndrome, and paraneoplastic syndromes; 0 13. cardiovascular: atherosclerosis, affecting the coronary and peripheral circulation; pericarditis; myocarditis , inflammatory and auto-immune cardiomyopathies including myocardial sarcoid; ischaemic reperfusion injuries; endocarditis, valvulitis, and aortitis including infective (for example syphilitic); vasculitides; disorders of the proximal and peripheral veins including phlebitis and thrombosis, including deep vein thrombosis and complications of varicose veins;

14. oncology: treatment of common cancers including prostate, breast, lung, ovarian, pancreatic, bowel and colon, stomach, skin and brain tumors and malignancies affecting the bone marrow (including the leukaemias) and lymphoproliferative systems, such as Hodgkin's and non-Hodgkin's lymphoma; including the prevention and treatment of metastatic disease and tumour recurrences, and paraneoplastic syndromes; and,

15. gastrointestinal tract: Coeliac disease, proctitis, eosinopilic gastro-enteritis, mastocytosis, Crohn's disease, ulcerative colitis, microscopic colitis, indeterminant colitis, irritable bowel disorder, irritable bowel syndrome, non-inflammatory diarrhea, food- related allergies which have effects remote from the gut, e.g., migraine, rhinitis and eczema.

Accordingly, the present invention further provides a compound of the present invention, as hereinbefore defined, for use in therapy.

In another aspect, the invention provides the use of a compound of the present invention, as hereinbefore defined, in the manufacture of a medicament for use in therapy.

In the context of the present specification, the term "therapy" also includes "prophylaxis" unless there are specific indications to the contrary. The terms "therapeutic" and "therapeutically" should be construed accordingly.

The present invention also provides a compound of the present invention, as hereinbefore defined, for treating chronic obstructive pulmonary disease (COPD) (such as irreversible COPD).

The present invention also provides a compound of the present invention, as hereinbefore defined, for treating asthma. A further aspect of the invention provides a method of treating a disease state in a mammal suffering from, or at risk of, said disease, which comprises administering to a mammal in need of such treatment a therapeutically effective amount of a compound of the present invention, as hereinbefore defined.

The present invention also provides the use of a compound of the present invention, as hereinbefore defined, in the manufacture of a medicament for use in the treatment of chronic obstructive pulmonary disease (COPD) (such as irreversible COPD).

The present invention also provides the use of a compound of the present invention as hereinbefore defined, in the manufacture of a medicament for use in the treatment of asthma.

The present invention further provides a method of treating chronic obstructive pulmonary disease (COPD) (such as irreversible COPD), in a warm-blooded animal, such as man, which comprises administering to a mammal in need of such treatment an effective amount of a compound of the present invention, as hereinbefore defined.

The present invention further provides a method of treating asthma in a warm-blooded animal, such as man, which comprises administering to a mammal in need of such treatment an effective amount of a compound of the present invention, as hereinbefore defined.

In order to use a compound of the invention for the therapeutic treatment of a warm- blooded animal, such as man, said ingredient is normally formulated in accordance with standard pharmaceutical practice as a pharmaceutical composition.

Therefore in another aspect the present invention provides a pharmaceutical composition that comprises a compound of the invention, as hereinbefore defined, and a pharmaceutically acceptable adjuvant, diluent or carrier. In a further aspect the present invention provides a process for the preparation of said composition, which comprises mixing active ingredient with a pharmaceutically acceptable adjuvant, diluent or carrier. Depending on the mode of administration, the pharmaceutical composition will, for example, comprise from 0.05 to 99%w (per cent by weight), such as from 0.05 to 80%w, for example from 0.10 to 70%w, such as from 0.10 to 50%w, of active ingredient, all percentages by weight being based on total composition.

The pharmaceutical compositions of this invention may be administered in standard manner for the disease condition that it is desired to treat, for example by topical (such as to the lung and/or airways or to the skin), oral, rectal or parenteral administration. For these purposes the compounds of this invention may be formulated by means known in the art into the form of, for example, aerosols, dry powder formulations, tablets, capsules, syrups, powders, granules, aqueous or oily solutions or suspensions, (lipid) emulsions, dispersible powders, suppositories, ointments, creams, drops and sterile injectable aqueous or oily solutions or suspensions.

A suitable pharmaceutical composition of this invention is one suitable for oral administration in unit dosage form, for example a tablet or capsule, which contains between 0. lmg and Ig of active ingredient.

In another aspect a pharmaceutical composition of the invention is one suitable for intravenous, subcutaneous or intramuscular injection. Each patient may receive, for example, an intravenous, subcutaneous or intramuscular dose of O.Olmgkg^"1 to lOOmgkg^"1 of the compound, for example in the range of 0. lmgkg^"1 to 20mgkg^~I of this invention, the composition being administered 1 to 4 times per day. The intravenous, subcutaneous and intramuscular dose may be given by means of a bolus injection. Alternatively the intravenous dose may be given by continuous infusion over a period of time. Alternatively each patient will receive a daily oral dose, which is approximately equivalent to the daily parenteral dose, the composition being administered 1 to 4 times per day.

Another suitable pharmaceutical composition of this invention is one suitable for inhaled administration, inhalation being a particularly useful method for administering the compounds of the invention when treating respiratory diseases such as chronic obstructive pulmonary disease (COPD) or asthma. When administered by inhalation the compounds of the present invention may be used effectively at doses in the μg range, for example 0.1 to 500 μg, 0.1 to 50 μg, 0.1 to 40 μg, 0.1 to 30 μg, 0.1 to 20 μg, 0.1 to 10 μg, 5 to 10 μg, 5 to 50 μg, 5 to 40 μg, 5 to 30 μg, 5 to 20 μg, 5 to 10 μg, 10 to 50 μg, 10 to 40 μg 10 to 30 μg, or 10 to 20 μg of active ingredient.

In an embodiment of the invention, there is provided a pharmaceutical composition comprising a compound of the invention, as hereinbefore defined, in association with a pharmaceutically acceptable adjuvant, diluent or carrier, which is formulated for inhaled administration.

When administered by inhalation, metered dose inhaler devices may be used to administer the active ingredient, dispersed in a suitable propellant and with or without additional excipients such as ethanol, surfactants, lubricants or stabilising agents. Suitable propellants include hydrocarbon, chlorofluorocarbon and hydrofluoroalkane (e.g. heptafluoroalkane) propellants, or mixtures of any such propellants. Preferred propellants are P134a and P227, each of which may be used alone or in combination with other propellants and/or surfactant and/or other excipients. Nebulised aqueous suspensions or, preferably, solutions may also be employed, with or without a suitable pH and/or tonicity adjustment, either as a unit-dose or multi-dose formulations.

Dry powder inhalers may be used to administer the active ingredient, alone or in combination with a pharmaceutically acceptable carrier, in the later case either as a finely divided powder or as an ordered mixture. The dry powder inhaler may be single dose or multi-dose and may utilise a dry powder or a powder-containing capsule.

Metered dose inhaler, nebuliser and dry powder inhaler devices are well known and a variety of such devices are available.

The invention further relates to combination therapies wherein a compound of the invention or a pharmaceutical composition or formulation comprising a compound of the invention, is administered concurrently or sequentially or as a combined preparation with another therapeutic agent or agents, for the treatment of one or more of the conditions listed.

In particular, for the treatment of the inflammatory diseases such as (but not restricted to) rheumatoid arthritis, osteoarthritis, asthma, allergic rhinitis, chronic obstructive pulmonary disease (COPD), psoriasis, and inflammatory bowel disease, the compounds of the invention may be combined with agents listed below.

Non-steroidal anti-inflammatory agents (hereinafter NSAIDs) including non-selective cyclo-oxygenase COX-I / COX-2 inhibitors whether applied topically or systemically

(such as piroxicam, diclofenac, propionic acids such as naproxen, flurbiprofen, fenoprofen, ketoprofen and ibuprofen, fenamates such as mefenamic acid, indomethacin, sulindac, azapropazone, pyrazolones such as phenylbutazone, salicylates such as aspirin); selective COX-2 inhibitors (such as meloxicam, celecoxib, rofecoxib, valdecoxib, lumarocoxib, parecoxib and etoricoxib); cyclo-oxygenase inhibiting nitric oxide donors (CINODs); glucocorticosteroids (whether administered by topical, oral, intramuscular, intravenous, or intra-articular routes); methotrexate; leflunomide; hydroxychloroquine; d-penicillamine; auranofin or other parenteral or oral gold preparations; analgesics; diacerein; intra-articular therapies such as hyaluronic acid derivatives; and nutritional supplements such as glucosamine.

The present invention still further relates to the combination of a compound of the invention together with a cytokine or agonist or antagonist of cytokine function, (including agents which act on cytokine signalling pathways such as modulators of the SOCS system) including alpha-, beta-, and gamma-interferons; insulin-like growth factor type I (IGF-I); interleukins (IL) including ILl to 17, and interleukin antagonists or inhibitors such as anakinra; tumour necrosis factor alpha (TNF-α) inhibitors such as anti-TNF monoclonal antibodies (for example infliximab; adalimumab, and CDP-870) and TNF receptor antagonists including immunoglobulin molecules (such as etanercept) and low-molecular- weight agents such as pentoxyfylline. In addition the invention relates to a combination of a compound of the invention with a monoclonal antibody targeting B-Lymphocytes (such as CD20 (rituximab), MRA-aIL16R and T-Lymphocytes, CTLA4-Ig, HuMax 11-15).

The present invention still further relates to the combination of a compound of the invention with a modulator of chemokine receptor function such as an antagonist of CCRl, CCR2, CCR2A, CCR2B, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCRlO and CCRl 1 (for the C-C family); CXCRl, CXCR2, CXCR3, CXCR4 and CXCR5

(for the C-X-C family) and CX₃CRl for the C-X₃-C family.

The present invention further relates to the combination of a compound of the invention with an inhibitor of matrix metalloprotease (MMPs), i.e., the stromelysins, the collagenases, and the gelatinases, as well as aggrecanase; especially collagenase-1 (MMP- 1), collagenase-2 (MMP-8), collagenase-3 (MMP- 13), stromelysin-1 (MMP-3), stromelysin-2 (MMP-10), and stromelysin-3 (MMP-11) and MMP-9 and MMP-12, including agents such as doxycycline.

The present invention still further relates to the combination of a compound of the invention and a leukotriene biosynthesis inhibitor, 5-lipoxygenase (5-LO) inhibitor or 5- lipoxygenase activating protein (FLAP) antagonist such as; zileuton; ABT-761; fenleuton; tepoxalin; Abbott-79175; Abbott-85761; a N-(5-substituted)-thiophene-2- alkylsulfonamide; 2,6-di-tert-butylphenolhydrazones; a methoxytetrahydropyrans such as Zeneca ZD-2138; the compound SB-210661; a pyridinyl-substituted 2-cyanonaphthalene compound such as L-739,010; a 2-cyanoquinoline compound such as L-746,530; or an indole or quinoline compound such as MK-591, MK-886, and BAY x 1005.

The present invention further relates to the combination of a compound of the invention and a receptor antagonist for leukotrienes (LT) B4, LTC4, LTD4, and LTE4 selected from the group consisting of the phenothiazin-3-ls such as L-651,392; amidino compounds such as CGS-25019c; benzoxalamines such as ontazolast; benzenecarboximidamides such as BIIL 284/260; and compounds such as zafirlukast, ablukast, montelukast, pranlukast, verlukast (MK-679), RG-12525, Ro-245913, iralukast (CGP 45715A), and BAY x 7195. The present invention still further relates to the combination of a compound of the invention and a phosphodiesterase (PDE) inhibitor such as a methylxanthanine including theophylline and aminophylline; a selective PDE isoenzyme inhibitor including a PDE4 inhibitor an inhibitor of the isoform PDE4D, or an inhibitor of PDE5.

The present invention further relates to the combination of a compound of the invention and a histamine type 1 receptor antagonist such as cetirizine, loratadine, desloratadine, fexofenadine, acrivastine, terfenadine, astemizole, azelastine, levocabastine, chlorpheniramine, promethazine, cyclizine, or mizolastine; applied orally, topically or parenterally.

The present invention still further relates to the combination of a compound of the invention and a proton pump inhibitor (such as omeprazole) or a gastroprotective histamine type 2 receptor antagonist.

The present invention further relates to the combination of a compound of the invention and an antagonist of the histamine type 4 receptor.

The present invention still further relates to the combination of a compound of the invention and an alpha- l/alpha-2 adrenoceptor agonist vasoconstrictor sympathomimetic agent, such as propylhexedrine, phenylephrine, phenylpropanolamine, ephedrine, pseudoephedrine, naphazoline hydrochloride, oxymetazoline hydrochloride, tetrahydrozoline hydrochloride, xylometazoline hydrochloride, tramazoline hydrochloride or ethylnorepinephrine hydrochloride.

The present invention still further relates to the combination of a compound of the invention and a beta- adrenoceptor agonist (including beta receptor subtypes 1-4) such as isoprenaline, salbutamol, formoterol, salmeterol, terbutaline, orciprenaline, bitolterol mesylate, pirbuterol, or indacaterol or a chiral enantiomer thereof. The present invention further relates to the combination of a compound of the invention and a chromone, such as sodium cromoglycate or nedocromil sodium.

The present invention still further relates to the combination of a compound of the invention with a glucocorticoid, such as flunisolide, triamcinolone acetonide, beclomethasone dipropionate, budesonide, fluticasone propionate, ciclesonide or mometasone furoate.

The present invention further relates to the combination of a compound of the invention with an agent that modulates a nuclear hormone receptor such as PPARs.

The present invention still further relates to the combination of a compound of the invention together with an immunoglobulin (Ig) or Ig preparation or an antagonist or antibody modulating Ig function such as anti-IgE (for example omalizumab).

The present invention further relates to the combination of a compound of the invention and another systemic or topically-applied anti-inflammatory agent, such as thalidomide or a derivative thereof, a retinoid, dithranol or calcipotriol.

The present invention still further relates to the combination of a compound of the invention and combinations of aminosalicylates and sulfapyridine such as sulfasalazine, mesalazine, balsalazide, and olsalazine; and immunomodulatory agents such as the thiopurines, and corticosteroids such as budesonide.

The present invention further relates to the combination of a compound of the invention together with an antibacterial agent such as a penicillin derivative, a tetracycline, a macrolide, a beta-lactam, a fluoroquinolone, metronidazole, an inhaled aminoglycoside; an antiviral agent including acyclovir, famciclovir, valaciclovir, ganciclovir, cidofovir, amantadine, rimantadine, ribavirin, zanamavir and oseltamavir; a protease inhibitor such as indinavir, nelfinavir, ritonavir, and saquinavir; a nucleoside reverse transcriptase inhibitor such as didanosine, lamivudine, stavudine, zalcitabine or zidovudine; or a non-nucleoside reverse transcriptase inhibitor such as nevirapine or efavirenz. The present invention still further relates to the combination of a compound of the invention and a cardiovascular agent such as a calcium channel blocker, a beta- adrenoceptor blocker, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin-2 receptor antagonist; a lipid lowering agent such as a statin or a fibrate; a modulator of blood cell morphology such as pentoxyfylline; thrombolytic, or an anticoagulant such as a platelet aggregation inhibitor.

The present invention further relates to the combination of a compound of the invention and a CNS agent such as an antidepressant (such as sertraline), an anti-Parkinsonian drug (such as deprenyl, L-dopa, ropinirole, pramipexole, a MAOB inhibitor such as selegine and rasagiline, a comP inhibitor such as tasmar, an A-2 inhibitor, a dopamine reuptake inhibitor, an NMDA antagonist, a nicotine agonist, a dopamine agonist or an inhibitor of neuronal nitric oxide synthase), or an anti- Alzheimer's drug such as donepezil, rivastigmine, tacrine, a COX-2 inhibitor, propentofylline or metrifonate.

The present invention still further relates to the combination of a compound of the invention and an agent for the treatment of acute or chronic pain, such as a centrally or peripherally-acting analgesic (for example an opioid or derivative thereof), carbamazepine, phenytoin, sodium valproate, amitryptiline or other anti-depressant agent-s, paracetamol, or a non-steroidal anti-inflammatory agent.

The present invention further relates to the combination of a compound of the invention together with a parenterally or topically-applied (including inhaled) local anaesthetic agent such as lignocaine or a derivative thereof.

A compound of the present invention can also be used in combination with an anti- osteoporosis agent including a hormonal agent such as raloxifene, or a biphosphonate such as alendronate.

The present invention still further relates to the combination of a compound of the invention together with a: (i) tryptase inhibitor; (ii) platelet activating factor (PAF) antagonist; (iii) interleukin converting enzyme (ICE) inhibitor; (iv) IMPDH inhibitor; (v) adhesion molecule inhibitors including VLA-4 antagonist; (vi) cathepsin; (vii) kinase inhibitor such as an inhibitor of tyrosine kinase (such as Btk, Itk, Jak3 or MAP, for example Gefitinib or ϊmatinib mesylate), a serine / threonine kinase (such as an inhibitor of a MAP kinase such as p38, JNK, protein kinase A, B or C, or IKK), or a kinase involved in cell cycle regulation (such as a cylin dependent kinase); (viii) glucose-6 phosphate dehydrogenase inhibitor; (ix) kinin-B 1. - or B2. -receptor antagonist; (x) anti-gout agent, for example colchicine; (xi) xanthine oxidase inhibitor, for example allopurinol; (xii) uricosuric agent, for example probenecid, sulfinpyrazone or benzbromarone; (xiii) growth hormone secretagogue; (xiv) transforming growth factor (TGFβ); (xv) platelet-derived growth factor (PDGF); (xvi) fibroblast growth factor for example basic fibroblast growth factor (bFGF); (xvii) granulocyte macrophage colony stimulating factor (GM-CSF); (xviii) capsaicin cream; (xix) tachykinin NKl or NK3 receptor antagonist such as NKP-608C, SB-233412 (talnetant) or D-4418; (xx) elastase inhibitor such as UT-77 or ZD-0892; (xxi) TNF- alpha converting enzyme inhibitor (TACE); (xxii) induced nitric oxide synthase

(iNOS) inhibitor; (xxiii) chemoattractant receptor-homologous molecule expressed on TH2 cells, (such as a CRTH2 antagonist); (xxiv) inhibitor of P38; (xxv) agent modulating the function of Toll-like receptors (TLR), (xxvi) agent modulating the activity of purinergic receptors such as P2X7; or (xxvii) inhibitor of transcription factor activation such as NFkB, API, or STATS.

A compound of the invention can also be used in combination with an existing therapeutic agent for the treatment of cancer, for example suitable agents include:

(i) an antiproliferative/antineoplastic drug or a combination thereof, as used in medical oncology, such as an alkylating agent (for example cis-platin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulphan or a nitrosourea); an antimetabolite (for example an antifolate such as a fluoropyrimidine like 5-fluorouracil or tegafur, raltitrexed, methotrexate, cytosine arabinoside, hydroxyurea, gemcitabine or paclitaxel); an antitumour antibiotic (for example an anthracycline such as adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin or mithramycin); an antimitotic agent (for example a vinca alkaloid such as vincristine, vinblastine, vindesine or vinorelbine, or a taxoid such as taxol or taxotere); or a topoisomerase inhibitor (for example an epipodophyllotoxin such as etoposide, teniposide, amsacrine, topotecan or a camptothecin);

(ii) a cytostatic agent such as an antioestrogen (for example tamoxifen, toremifene, raloxifene, droloxifene or iodoxyfene), an oestrogen receptor down regulator (for example fulvestrant), an antiandrogen (for example bicalutamide, flutamide, nilutamide or cyproterone acetate), a LHRH antagonist or LHRH agonist (for example goserelin, leuprorelin or buserelin), a progestogen (for example megestrol acetate), an aromatase inhibitor (for example as anastrozole, letrozole, vorazole or exemestane) or an inhibitor of 5α-reductase such as finasteride; (iii) an agent which inhibits cancer cell invasion (for example a metalloproteinase inhibitor like marimastat or an inhibitor of urokinase plasminogen activator receptor function); (iv) an inhibitor of growth factor function, for example: a growth factor antibody (for example the anti-erbb2 antibody trastuzumab, or the anti-erbbl antibody cetuximab [C225]), a farnesyl transferase inhibitor, a tyrosine kinase inhibitor or a serine/threonine kinase inhibitor, an inhibitor of the epidermal growth factor family (for example an EGFR family tyrosine kinase inhibitor such as N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3- morpholinopropoxy)quinazolin-4-amine (gefitinib, AZD 1839), N-(3-ethynylphenyl)-6,7- bis(2-methoxyethoxy)quinazolin-4-amine (erlotinib, OSI-774) or 6-acrylamido~N-(3- chloro-4-fluorophenyl)-7-(3-morpholinopropoxy)quinazolin-4-amine (CI 1033)), an inhibitor of the platelet-derived growth factor family, or an inhibitor of the hepatocyte growth factor family;

(v) an antiangiogenic agent such as one which inhibits the effects of vascular endothelial growth factor (for example the anti-vascular endothelial cell growth factor antibody bevacizumab, a compound disclosed in WO 97/22596, WO 97/30035, WO 97/32856 or WO 98/13354), or a compound that works by another mechanism (for example linomide, an inhibitor of integrin αvβ3 function or an angiostatin);

(vi) a vascular damaging agent such as combretastatin A4, or a compound disclosed in WO 99/02166, WO 00/40529, WO 00/41669, WO 01/92224, WO 02/04434 or WO 02/08213; (vii) an agent used in antisense therapy, for example one directed to one of the targets listed above, such as ISIS 2503, an anti-ras antisense;

(viii) an agent used in a gene therapy approach, for example approaches to replace aberrant genes such as aberrant p53 or aberrant BRCAl or BRCA2, GDEPT (gene-directed enzyme pro-drug therapy) approaches such as those using cytosine deaminase, thymidine kinase or a bacterial nitroreductase enzyme and approaches to increase patient tolerance to chemotherapy or radiotherapy such as multi-drug resistance gene therapy; or (ix) an agent used in an immunotherapeutic approach, for example ex-vivo and in-vivo approaches to increase the immunogenicity of patient tumour cells, such as transfection with cytokines such as interleukin 2, interleukin 4 or granulocyte-macrophage colony stimulating factor, approaches to decrease T-cell anergy, approaches using transfected immune cells such as cytokine-transfected dendritic cells, approaches using cytokine-transfected tumour cell lines and approaches using anti-idiotypic antibodies.

In a further embodiment the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is a compound of the present invention, as hereinbefore described, and at least one further active ingredient selected from:- • a phosphodiesterase inhibitor

• a β2. adrenoceptor agonist

• a modulator of chemokine receptor function

• an inhibitor of kinase function

• a protease inhibitor • a steroidal glucocorticoid receptor agonist

• a non-steroidal glucocorticoid receptor agonist.

The pharmaceutical product according to this embodiment may, for example, be a pharmaceutical composition comprising the first and further active ingredients in admixture. Alternatively, the pharmaceutical product may, for example, comprise the first and further active ingredients in separate pharmaceutical preparations suitable for simultaneous, sequential or separate administration to a patient in need thereof. The pharmaceutical product of this embodiment is of particular use in treating respiratory diseases such as asthma, COPD or rhinitis. Examples of a phosphodiesterase inhibitor that may be used in the pharmaceutical product according to this embodiment include a PDE4 inhibitor such as an inhibitor of the isoform

PDE4D, a PDE3 inhibitor and a PDE5 inhibitor. Examples include the compounds

(Z)-3-(3,5-dichloro-4-pyridyl)-2-[4-(2-indanyloxy-5-methoxy-2-pyridyl]propenenitrile, N-[9-amino-4-oxo-l-phenyl-3,4,6,7-tetrahydropyrrolo[3,2,l-jk][l,4]benzodiazepin-3(R)- yl]pyridine-3-carboxamide (CI- 1044)

3-(benzyloxy)-l-(4-fluorobenzyl)-N-[3-(methylsulphonyl)phenyl]-lH-indole-2- carboxamide,

(lS-exo)-5-[3-(bicyclo[2.2.1]hept-2-yloxy)-4-methoxyphenyl]tetrahydro-2(lH)- pyrimidinone (Atizoram),

N-(3,5,dichloro-4-pyridinyl)-2-[l-(4-fluorobenzyl)-5-hydroxy-lH-indol-3-yl]-2- oxoacetamide (AWD- 12-281), β-[3-(cyclopentyloxy)-4-methoxyphenyl]- 1 ,3-dihydro- 1 ,3-dioxo-2H-isoindole-2- propanamide (CDC-801), N-[9-methyl-4-oxo-l-phenyl-3,4,6,7-tetrahydropyrrolo[3,2,l-jk][l,4]benzodiazepin-3(R)- yl]pyridine-4-carboxamide (CI- 1018), cis-[4-cyano-4-(3-cyclopentyloxy-4-methoxyphenyl)cyclohexane- 1 -carboxylic acid

(Cilomilast)

8-amino- 1 ,3-bis(cyclopropylmethyl)xanthine (Cipamfylline) N-(2,5-dichloro-3-pyridinyl)-8-methoxy-5-quinolinecarboxamide (D-4418),

5-(3,5-di-tert-butyl-4-hydroxybenzylidene)-2-iminothiazolidin-4-one (Darbufelone),

2-methyl-l-[2-(l-methylethyl)pyrazolo[l,5-a]pyridin-3-yl]-l-propanone (Ibudilast),

2-(2,4-dichlorophenylcarbonyl)-3-ureidobenzofuran-6-yl methanesulphonate (Lirimilast),

(-)-(R)-5-(4-methoxy-3-propoxyphenyl)-5-methyloxazolidin-2-one (Mesopram), (-)-cis-9-ethoxy-8-methoxy-2-methyl- 1,2,3, 4,4a, 10b-hexahydro-6-(4- diisopropylaminocarbonylphenyl)-benzo[c] [ 1 ,6]naphthyridine (Pumafen trine),

3-(cyclopropylmethoxy)-N-(3,5-dichloro-4-pyridyl)-4-(difluoromethoxy)benzamide

(Roflumilast), the N-oxide of Roflumilast, 5,6-diethoxybenzo[b]thiophene-2-carboxylic acid (Tibenelast)

2,3,6,7-tetrahydro-2-(mesitylimino)-9,10-dimethoxy-3-methyl-4H-pyrimido[6,l- a]isoquinolin-4-one (trequinsin) and 3-[[3-(cycloρentyloxy)-4-methoxyphenyl]-methyl]-N-ethyl-8-(l-methylethyl)-3H-purine- 6-amine (V-11294A).

Examples of a β₂-adrenoceptor agonist that may be used in the pharmaceutical product according to this embodiment include metaproterenol, isoproterenol, isoprenaline, albuterol, salbutamol (e.g. as sulphate), formoterol (e.g. as fumarate), salmeterol (e.g. as xinafoate), terbutaline, orciprenaline, bitolterol (e.g. as mesylate), pirbuterol or indacaterol. The β₂-adrenoceptor agonist of this embodiment may be a long-acting β₂-agonists, for example salmeterol (e.g. as xinafoate), formoterol (e.g. as fumarate), bambuterol (e.g. as hydrochloride), carmoterol (TA 2005, chemically identified as 2(1H)-Quinolone, 8- hydroxy-5-[l-hydroxy-2-[[2-(4-methoxy-phenyl)-l-methylethyl]-amino]ethyl]- monohydrochloride, [R-(R*, R*)] also identified by Chemical Abstract Service Registry Number 137888-11-0 and disclosed in U.S. Patent No 4,579,854), indacaterol (CAS no 312753-06-3; QAB-149), formanilide derivatives e.g. 3-(4-{ [6-({ (2R)-2-[3- (formylamino)-4-hydroxyphenyl]-2-hydroxyethyl } amino)hexyl]oxy } -butyl)- benzenesulfonamide as disclosed in WO 2002/76933, benzenesulfonamide derivatives e.g. 3-(4- { [6-( { (2R)-2-hydroxy-2-[4-hydroxy-3-(hydroxy-methyl)phenyl]ethyl } amino)- hexyl]oxy} butyl )benzenesulfonamide as disclosed in WO 2002/88167, aryl aniline receptor agonists as disclosed in WO 2003/042164 and WO 2005/025555, indole derivatives as disclosed in WO 2004/032921, in US 2005/222144, compounds GSK 159797, GSK 159802, GSK 597901, GSK 642444 and GSK 678007.

Examples of a modulator of chemokine receptor function that may be used in the pharmaceutical product according to this embodiment include a CCRl receptor antagonist.

Examples of an inhibitor of kinase function that may be used in the pharmaceutical product according to this embodiment include a p38 kinase inhibitor and an IKK inhibitor.

Examples of a protease inhibitor that may be used in the pharmaceutical product according to this embodiment include an inhibitor of neutrophil elastase or an inhibitor of MMP 12. Examples of a steroidal glucocorticoid receptor agonist that may be used in the pharmaceutical product according to this embodiment include budesonide, fluticasone (e.g. as propionate ester), mometasone (e.g. as furoate ester), beclomethasone (e.g. as 17- propionate or 17,21-dipropionate esters), ciclesonide, loteprednol (as e.g. etabonate), etiprednol (as e.g. dicloacetate), triamcinolone (e.g. as acetonide), flunisolide, zoticasone, flumoxonide, rofleponide, butixocort (e.g. as propionate ester), prednisolone, prednisone, tipredane, steroid esters e.g. 6α,9α-difluoro-17α-[(2-furanylcarbonyl)oxy]-l lβ-hydroxy- 16α-methyl-3-oxo-androsta-l,4-diene-17β-carbothioic acid S-fluoromethyl ester, 6α,9α- difluoro- 11 β-hydroxy- 16α-methyl-3-oxo- 17α-propionyloxy-androsta- 1 ,4-diene- 17β- carbothioic acid S-(2-oxo-tetrahydro-furan-3S-yl) ester and 6α,9α-difiuoro-llβ-hydroxy- 16α-methyl- 17α-[(4-methyl- 1 ,3-thiazole-5-carbonyl)oxy]-3-oxo-androsta- 1 ,4-diene- 17β- carbothioic acid S-fluoromethyl ester, steroid esters according to DE 4129535 , steroids according to WO 2002/00679, WO 2005/041980, or steroids GSK 870086, GSK 685698 and GSK 799943.

Examples of a modulator of a non-steroidal glucocorticoid receptor agonist that may be used in the pharmaceutical product according to this embodiment include those described in WO2006/046916.

The present invention will now be illustrated with the following non-limiting Examples.

In the examples the NMR spectra were measured on a Varian Unity Inova spectrometer at a proton frequency of either 300, 400 or 500 MHz. The MS spectra were measured on either an Agilent 1100 MSD G1946D spectrometer or a Hewlett Packard HPl 100 MSD G 1946 A spectrometer. Preparative HPLC separations were performed using a Waters

Symmetry^® or Xterra^® column using 0.1% aqueous trifluoroacetic acid: acetonitrile, 0.1% aqueous ammonia: acetonitrile or 0.1% ammonium acetate: acetonitrile as the eluent. Preparative Chiral HPLC separations were performed by the systems listed in the examples and chiral purity of the resulting fragments confirmed to be >95% enantiomeric excess by analytical Chiral HPLC. SCX and NH₂ resin were obtained from Varian Incorporated. IUPAC names were generated using Beilstein Autonom 2000. Stereochemistry was assigned according to the Cahn-Ingold-Prelog system. Absolute configuration at the 3 position is assigned on the basis of the (i?)-quinuclidin-3-ol intermediate; (2?)-quinuclidin- 3-ol is commercially available from Acros Organics. Absolute configuration at the 2' position of Examples 1, 3, 4, 5, 6, 7, 8, 9, 10, 11, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32 and 33 is assigned on the basis of the (,S)-2-Phenyl-2-ρiperidin-l-yl- propionic acid methyl ester intermediate (Intermediate A - Isomer 1) used in their preparation. The absolute configuration of Intermediate A - Isomer 1 is assigned on the basis of the absolute configuration of Reference Compound 1 , which was assigned by single crystal X-ray diffraction. The preparation of Intermediate A - Isomer 1, its use in the preparation of Reference Compound 1, and the assignment of the absolute configuration of Reference Compound 1 by single crystal X-ray diffraction is described below.

Intermediate A (Isomers 1 & 2): 2-Phenyl-2-piperidin-l-yl-propionic acid methyl ester

A solution of methyl 2-bromo-2-phenylpropanoate (1 g) in acetonitrile (30 mL) was treated with piperidine (1 mL). The solution was stirred and heated under reflux for 3 h then concentrated to dryness. The residue was purified by flash column chromatography on silica gel using ether / isohexane (3:7) to afford the racemic sub-titled compound as a colourless oil (0.8 g). The mixture of enantiomers was separated by chiral hplc using a chiracel OJ-H column using an isocratic system of 80% isohexane / ethanol to afford the two enantiomers, which were defined as Isomer 1 and Isomer 2 in order of elution.

2-Phenyl-2-piperidin-l-yl-propionic acid methyl ester (Isomer 1)

Chiral HPLC 80:20 isohexane : ethanol (isocratic). Chiracel OJ-H 4.6mm x 50mm Retention time 1.09min. ¹H NMR (400 MHz, CDCl₃) δ 7.56 - 7.49 (2H, m), 7.35 - 7.20 (3H, m), 3.68 (3H, s), 2.54 2.45 (2H, m), 2.41 - 2.32 (2H, m), 1.64 - 1.54 (7H, m), 1.50 - 1.42 (2H, m).

2-Phenyl-2-piρeridin-l-yl-propionic acid methyl ester (Isomer 2)

Chiral HPLC 80:20 isohexane : ethanol (isocratic). Chiracel OJ-H 4.6mm x 50mm Retention time 2.52min.

¹H NMR (400 MHz, CDCl₃) δ 7.56 - 7.49 (2H, m), 7.35 - 7.20 (3H, m), 3.68 (3H, s), 2.54 - 2.45 (2H, m), 2.41 - 2.32 (2H, m), 1.64 - 1.54 (7H, m), 1.50 - 1.42 (2H, m).

Reference Compound 1: (R)-l-(2-Benzyloxy-ethyl)-3-((S)-2-phenyl-2-piperidin-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

Intermediate B (Isomer 1): 2-Phenyl-2-piperidin-l-yl-proρionic acid (i?)-(l~aza- bicyclo[2.2.2]oct-3-yl) ester

A mixture of 2-phenyl-2-piperidin- 1-yl-propionic acid methyl ester (Intermediate A,

Isomer 1) (0.9 g), (i?)-quinuclidin-3-ol (1.157 g) and sodium hydride (60% in mineral oil, 0.335 g) in dry toluene (20 mL) was heated at 12O⁰C under an atmosphere of nitrogen for 8h. The cooled reaction mixture was diluted with water (100 mL) and extracted with diethyl ether (2 x 150 mL). The combined extracts were dried (MgSO₄) and concentrated to give an oil. The crude product was purified by flash column chromatography on silica eluting with ethyl acetate / methanol 9:1 to afford the titled compound (0.500 g).

¹H NMR (400 MHz, DMSO) δ 7.59 - 7.51 (2H, m), 7.40 - 7.21 (3H, m), 4.72 - 4.62 (IH, m), 3.34 - 3.26 (IH, m), 3.04 - 2.92 (IH, m), 2.75 - 2.13 (7H, m), 1.89 - 1.75 (IH, m), 1.71 - 1.20 (14H, m).

2-Phenyl-2-piperidin-l-yl-propionic acid (/?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Intermediate B, Isomer 1) (75 mg) in acetonitrile (1 mL) was treated with (2-bromo- ethoxymethyl)-benzene (47 mg) and the mixture was heated to reflux for 2 minutes. The mixture was cooled and diethyl ether (3 mL) was added and the resulting solid was collected by filtration, washed with diethyl ether and dried to afford the titled compound (63 mg).

m/e 477 [M]⁺

¹H NMR (400 MHz, DMSO) δ 7.55 - 7.50 (2H, m), 7.41 - 7.23 (8H, m), 5.10 - 5.04 (IH, m), 4.50 (2H, s), 3.99 - 3.90 (IH, m), 3.88 - 3.77 (2H, m), 3.59 - 3.36 (5H, m), 3.30 - 3.18 (2H, m), 2.46 - 2.30 (4H, m), 2.23 - 2.18 (IH, m), 2.02 - 1.75 (4H, m), 1.78 (3H, s), 1.55 - 1.47 (4H, m), 1.46 - 1.38 (2H, m).

Single crystal X-ray diffraction data obtained for Reference Compound 1 proved the structure to be (i?)-l-(2-benzyloxy-ethyl)-3-((6')-2-phenyl-2-piperidin-l-yl-propionyloxy)- l-azonia-bicyclo[2.2.2]octane bromide. The data set was collected at RT with graphite monochromatized MoK(a) radiation on a KappaCCD Single-Crystal X-Ray diffractometer equipped with an k-axis goniometer and a CCD area detector (Nonius, 1998). The diffraction raw data were processed within the Denzo-SMN program package (Otwinowski & Minor, 1998) converting the information from the digital image frame to a file containing h, k, 1 indices, background and Lp corrected intensities of the diffraction spots, along with estimate of errors. On the basis of the crystal structure determined for Reference Compound 1, the absolute configuration of Intermediate A - Isomer 1 used in the preparation of Examples 1, 3, 4, 5, 6, 7, 8, 9, 10, 11, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32 and 33 has been assigned as (S)-2-Phenyl-2-piperidin-l-yl-propionic acid methyl ester.

Example 1: (/?)-l-[2-(2-FIuoro-phenyl)-ethyI]-3-((S)-2-phenyI-2"piperidin-l-yl- propionyloxy)-l-azonia-bicydo[2.2.2]octane bromide

a) (ιS')-2-Phenyl-2-piperidin-l-yl-propionic acid (7?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester

A mixture of (5)-2-Phenyl-2-piperidin-l-yl-propionic acid methyl ester (Intermediate A - Isomer 1) (0.9 g), (R)-quinuclidin~3-ol (1.157 g) and sodium hydride (60% in mineral oil, 0.335 g) in dry toluene (20 mL) was heated at 12O⁰C under an atmosphere of nitrogen for 8h. The cooled reaction mixture was diluted with water (100 mL) and extracted with diethyl ether (2 x 150 mL). The combined extracts were dried (MgSO₄) and concentrated to give an oil. The crude product was purified by flash column chromatography on silica eluting with ethyl acetate / methanol 9:1 to afford the titled compound (0.500 g).

¹H NMR (400 MHz, DMSO) δ 7.59 - 7.51 (2H, m), 7.40 - 7.21 (3H, m), 4.72 - 4.62 (IH, m), 3.34 - 3.26 (IH, m), 3.04 - 2.92 (IH, m), 2.75 - 2.13 (7H, m), 1.89 - 1.75 (IH, m), 1.71 - 1.20 (14H, m).

Example 1: (J?)-l-[2-(2-FIuoro-phenyI)-ethyl]-3-((5)-2-phenyl-2-piperidm-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

(S)-2-Phenyl-2-piperidin-l-yl-propionic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (150 mg) and l-(2-bromo-ethyl)-2-fluoro-benzene (89 mg) in acetonitrile (1 mL) were stirred at RT for 17h. The resulting suspension was diluted with diethyl ether (3 mL) and the solid collected by filtration to afford the titled compound as a colourless solid (163 mg).

m/e 465 [M]⁺

¹H NMR (400 MHz, DMSO) δ 7.59 - 7.54 (2H, m), 7.45 - 7.32 (4H, m), 7.31 - 7.18 (3H, m), 5.13 - 5.08 (IH, m), 4.00 - 3.91 (IH, m), 3.65 - 3.39 (5H, m), 3.28 - 3.19 (2H, m), 3.06 - 2.98 (2H, m), 2.47 - 2.32 (4H, m), 2.25 - 2.18 (IH, m), 2.05 - 1.70 (4H, m), 1.58 (3H, s), 1.56 - 1.49 (4H, m), 1.48 - 1.39 (2H, m).

Example 2: (i?)-l-[2-(2-Fluoro-phenyl)-ethyl]-3-(2-piperidin-l-yI-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide (Isomer 1)

a) 2-Piperi din- l-yl-2-thiophen-2-yl -propionic acid ethyl ester (Isomers 1 and 2)

To a solution under nitrogen of thionyl chloride (18.4 niL) in diethyl ether (180 mL) at O⁰C was added dropwise over 25min a solution of 2-hydroxy-2-thiophen-2-yl-proρionic acid ethyl ester (25.3 g) and pyridine (20.4 mL) in diethyl ether (180 mL). The suspension was stirred at O⁰C for a further 15min then diluted with 2M aq. HCl (250 mL) at <20°C. The aqueous phase was separated and extracted with ethyl acetate (2 x 100 mL). The combined organic extracts were washed with 20% aq. brine solution (200 mL), dried (sodium sulphate), filtered and evaporated. The residue was dissolved in NMP (120 mL) and treated dropwise at 0⁰C with a solution of piperidine (25 mL) in NMP (60 mL) over 5min. Reaction mixture was stirred at 0⁰C for lhr then allowed to warm to ambient and stirred for 16h. The reaction mixture was added to water (1.8 L) and 2M aq. HCl (200 mL) and the aqueous (pHl) phase was washed with ethyl acetate (3 x 250 mL). The aqueous phase was basified to pH12 with 48% aq. NaOH solution and extracted with ethyl acetate (2 x 300 mL); the combined organics were washed with 20% aq. brine solution (2 x 150 mL), dried (sodium sulphate), filtered and evaporated to give 23 g of a dark oil. Purification by flash chromatography on silica eluting with zrøhexane / ethyl acetate (90 : 10). Evaporation of eluent (azeotroping with acetonitrile to remove residual ethyl acetate) gave 2Og of a pale yellow liquid. The racemic mixture was separated by chiral hlpc using a Chiralpak OJ-H 50 x 250mm column eluting with 100% ethanol at a flow rate of 118mL/min and at 25⁰C to provide chirally pure material. The first eluting enantiomer was designated Isomer 1 , and the second eluting Isomer 2.

Isomer 1 m/e 268 [M+H]⁺

¹H NMR (300 MHz, CDCl₃) δ 7.24 (d, IH, d), 7.00 (IH, d), 6.90 (IH, t), 4.19 (2H, q), 2.53 (2H, quintet), 2.41 (2H, quintet), 1.64 (s, 3H, s), 1.62 - 1.54 (5H, m), 1.46 (2H, q), 1.25 (3H, t).

Isomer 2 m/e 268 [M+H]^{+ 1}H NMR (300 MHz, CDCl₃) δ 7.24 (IH, d), 7.00 (IH, d), 6.90 (IH, t), 4.19 (2H, q), 2.53 (2H, quintet), 2.41 (2H, quintet), 1.64 (s, 3H, s), 1.62 - 1.54 (5H, m), 1.46 (2H, q), 1.25 (3H, t).

b) 2-Piperidin-l-yl-2-thiophen-2-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Isomer 1)

Prepared by the method of Example Ia using 2-piperidin-l-yl-2-thiophen-2-yl-propionic acid ethyl ester (Isomer 1) (Example 2a).

¹H NMR (400 MHz, CDCl₃) δ 7.25 (IH, s), 7.02 - 6.99 (IH, m), 6.92 - 6.88 (IH, m), 4.83 4.78 (IH, m), 3.18 - 3.09 (IH, m), 2.89 - 2.65 (4H, m), 2.60 - 2.49 (3H, m), 2.45 - 2.34 (2H, m), 2.01 - 1.77 (2H, m), 1.74 - 1.33 (12H, m).

Example 2: (R)-l-[2-(2-FIuoro-phenyl)-ethyl]-3-(2-piperidin-l-yl-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide (Isomer 1)

2-Piperidin-l-yl-2-thiophen-2-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Isomer 1) (Example 2b) (340 mg) in acetonitrile (3 mL) was treated with l-(2-bromo- ethyl)-2-fluoro-benzene (238 mg) and the mixture stirred at room temperature for 48h. Diethyl ether (10 mL) was added and the mixture stirred for 30 minutes. The resulting solid was collected by filtration and recrystallised from acetonitrile (2 mL) to afford the titled compound as a solid (250 mg)

m/e 471 [M]⁺

¹H NMR (400 MHz, DMSO) δ 7.50 (IH, dd), 7.43 - 7.32 (2H, m), 7.26 - 7.18 (2H, m). 7.10 (IH, dd), 6.96 (IH, dd), 5.17 - 5.11 (IH, m), 4.00 - 3.92 (IH, m), 3.68 - 3.58 (IH, m), 3.57 - 3.39 (4H, m), 3.37 - 3.28 (IH, m), 3.26 - 3.20 (IH, m), 3.06 - 3.00 (2H, m), 2.49 - 2.43 (2H, m), 2.41 - 2.32 (2H, m), 2.31 - 2.24 (IH, m), 2.05 - 1.83 (4H, m), 1.62 (3H, s), 1.55 - 1.47 (4H, m), 1.47 - 1.39 (2H, m).

Example 3: (R)-l-(6-Methyl-benzooxazol-2-ylmethyl)-3-((S)-2-phenyl-2-piperidm-l- yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloromethyl-6-methyl-benzooxazole

To 2-amino-5-methyl-phenol (2 g) in acetic acid (20 niL) and was added 2-chloro- 1,1,1 - trimethoxy-ethane (3.01 g) and the mixture heated under reflux for 24h. The solvent was removed under reduced pressure and the residue treated with cone, sulfuric acid (5 mL). The brown solution was heated a 100⁰C for 2h, cooled to room temperature and quenched into water (200 mL). The mixture was made basic by the addition of solid potassium carbonate and the products extracted into diethyl ether (2 x 150 mL). The dried extracts were concentrated to an oil and the oil purified by flash column chromatography using diethyl ether / rsøhexane (3 : 7) to afford the sub-titled compound as an oil (0.170 g).

¹H NMR (400 MHz, DMSO) δ 7.60 (IH, d), 7.35 (IH, q), 7.18 (IH, dt), 4.74 (2H, s), 2.50 (3H, s).

Example 3: (i?)-l-(6-Methyl-benzooxazol-2-ylmethyl)-3-((S)-2-phenyl-2-piperidin-l- yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

A mixture of (S^-phenyl^-piperidin-l-yl-propionic acid (7?)-(l-aza-bicyclo[2.2.2]oct-3- yl) ester (Example Ia) (120 mg) and 2-chloromethyl-6-methyl-benzooxazole (89 mg) in acetonitrile (2 mL) was heated under reflux for 5 minutes. The solution was cooled to ambient temperature and the solution diluted with diethyl ether (20 mL). The resulting solid was collected by filtration and dried to afford the titled compound as a solid (105 mg).

m/e 488 [M]⁺

¹H NMR (400 MHz, DMSO) 67.67 (IH, d), 7.54 - 7.49 (3H, m), 7.33 (IH, d), 7.13 (2H, t), 7.04 (IH, t), 5.16 - 5.10 (IH, m), 4.76 (2H, dd), 4.02 (IH, ddd), 3.73 - 3.53 (3H, m), 3.21 - 3.11 (IH, m), 2.53 (3H, s), 2.48 - 2.39 (2H, m), 2.39 - 2.30 (3H, m), 2.17 - 2.05 (IH, m), 2.05 - 1.86 (3H, m), 1.59 (3H, s), 1.58 - 1.50 (4H, m), 1.48 - 1.40 (2H, m).

Example 4: (jR)-l-(3-Benzooxazol-2-yl-propyl)-3-((S)-2-phenyl-2-piperidin-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-(3-Bromo-propyl)-benzooxazole

2-Amino-phenol (1.09 g) in acetic acid (20 mL) was stirred and heated under reflux with 4- bromo-l,l,l-trimethoxy-butane (2.72 g) for 2h. The cooled reaction mixture was diluted with water (150 mL) and basified with sodium bicarbonate. The mixture was extracted with diethyl ether (2 x 150 mL) and the combined extracts dried (MgSO-O. Concentration of the diethyl ether gave a brown oil which was purified on silica eluting with diethyl ether / zjchexane (3 : 7) to afford the sub-titled compound as a yellow solid (0.5 g).

¹H NMR (400 MHz, CDCl₃) 87.71 - 7.64 (IH, m), 7.53 - 7.45 (IH, m), 7.35 - 7.28 (2H, m), 3.58 (2H, t), 3.14 (2H, t), 2.46 (2H, quintet). Example 4: (i?)-l-(3-Benzooxazol-2-yI-propyl)-3-((S)-2-phenyI-2-piperidin-l-yI- propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

A mixture of (5)-2-phenyl-2-piperidin-l-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3- yl) ester (Example Ia) (170 mg) and 2-(3-bromo-propyl)-benzooxazole (Example 4a) (143 mg) in acetonitrile (1 mL) was heated under reflux for 5 minutes. The cooled reaction mixture was treated with diethyl ether (3 mL) and the resulting precipitate collected by filtration to afford the titled compound as a solid (180 mg).

m/e 502 [M]⁺

¹H NMR (400 MHz, DMSO) 57.73 - 7.68 (2H, m), 7.57 - 7.53 (2H, m), 7.42 - 7.33 (4H, m), 7.27 - 7.22 (IH, m), 5.10 - 5.05 (IH, m), 3.91 - 3.84 (IH, m), 3.55 - 3.32 (5H, m), 3.18 - 3.08 (2H, m), 3.01 (2H, t), 2.47 - 2.38 (2H, m), 2.39 - 2.31 (2H, m), 2.23 - 2.11 (3H, m), 2.00 - 1.75 (4H, m), 1.56 (3H, s), 1.55 - 1.48 (4H, m), 1.47 - 1.39 (2H, m).

Example 5: (i?)-l-(5-ChIoro-benzooxazol-2-ylmethyI)-3-((5')-2-phenyl-2-piperidin-l- yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloromethyl-5-chloro-benzooxazole

2-Amino-4-chloro-phenol (2.5 g) in acetic acid (10 mL) and toluene (20 mL) was stirred and heated under reflux with 2-chloro-l,l,l-trimethoxy-ethane (3.23 g) for 17h. The cooled reaction mixture precipitated a solid which was removed by filtration. The filtrate was concentrated to dryness and the residue purified by flash column chromatography on silica using diethyl ether / isohexane (3 : 7) as eluant to afford the sub-titled compound as an orange solid (0.150 g).

¹H NMR (400 MHz, CDCl₃) δ 7.73 (IH, d), 7.48 (IH, d), 7.37 (IH, dd), 4.74 (2H, s).

Example 5: (i?)-l-(5-ChIoro-benzooxazol-2-ylmethyl)-3-((S)-2-phenyI-2-piperidin-l- yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

(S)-2-Phenyl-2-piperidin-l-yl -propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (220 mg) and 2-chloromethyl-5-chloro-benzooxazole (Example 5a) (130 mg) in acetonitrile (2 mL) were heated to reflux for 5 minutes. To the cooled solution was added diethyl ether (3 mL) and the resulting precipitate collected by filtration and dried at 50⁰C for 2h. The crude solid was recrystallised from acetone to afford the titled compound as a solid (270 mg).

m/e 508 [M]⁺

¹H NMR (400 MHz, DMSO) 6 8.05 (IH, d), 7.92 (IH, d), 7.62 (IH, dd), 7.52 (2H, d), 7.28 (2H, t), 7.19 (IH, t), 5.09 - 5.03 (IH, m), 4.99 (2H, dd), 4.17 - 4.08 (IH, m), 3.78 - 3.68 (IH, m), 3.66 - 3.53 (2H, m), 3.51 - 3.44 (IH, m), 3.42 - 3.35 (IH, m), 2.45 - 2.37 (2H, m), 2.37 - 2.29 (2H, m), 2.25 - 2.19 (IH, m), 2.02 - 1.77 (4H, m), 1.56 (3H, s), 1.54 - 1.46 (4H, m), 1.44 - 1.36 (2H, m).

Example 6: (/?)-l-(6-Chloro-benzooxazoI-2-yImethyI)-3-((5)-2-phenyI-2-piperidin-l- yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 6-Chloro-2-chloromethyl-benzooxazole

To 2-amino-5-chloro-phenol (2.5 g) in acetic acid (25 mL) was added 2-chloro- 1,1,1 - trimethoxy-ethane (3.23 g) and the mixture heated under reflux for Ih. The solvent was removed under vacuum and the residue purified by chromatography on silica gel using diethyl ether / isohexane (1 : 10) to afford the sub-titled compound as a solid (2.60 g).

¹H NMR (400 MHz, CDCl₃) δ 7.58 (IH, d), 7.36 (IH, dd), 7.66 (IH, d), 4.74 (2H, s).

Example 6: (R)-l-(6-Chloro-benzooxazol-2-ylmethyl)-3-((5')-2-phenyl-2-piperidin-l- yl-propionyloxy)-l-azonia-bicydo[2.2.2]octane chloride

(S)-2-Phenyl~2-piperidin-l-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (140 mg) in acetonitrile (2 mL) was treated with 6-chloro-2-chloromethyl- benzooxazole (Example 6a) (99 mg). The mixture was heated under reflux for 5 minutes, cooled and diethyl ether (3 mL) added. The resulting precipitate was collected by filtration to afford the titled compound as a solid (180 mg).

m/e 508 [M]⁺

¹H NMR (400 MHz, DMSO) δ 8.11 (IH, d), 7.92 (IH, d), 7.57 (IH, dd), 7.52 (2H, d), 7.31 - 7.24 (2H, m), 7.19 (IH, t), 5.09 - 5.03 (IH, m), 4.99 (2H, dd), 4.18 - 4.08 (IH, m), 3.78 - 3.69 (IH, m), 3.66 - 3.52 (2H, m), 3.53 - 3.44 (IH₅ m), 3.45 - 3.36 (IH, m), 2.46 - 2.38 (2H, m), 2.38 - 2.29 (2H, m), 2.25 - 2.19 (IH, m), 2.01 - 1.77 (4H, m), 1.56 (3H, s), 1.54 - 1.45 (4H, m), 1.45 - 1.36 (2H, m).

Example 7: (i?)-l-Benzooxazol-2-yImethyI-3-((5)-2-phenyl-2-piperidin-l-yl- propionyloxy)-! -azonia-bicy clo[2.2.2]octane chloride

A mixture of (S)-2-phenyl-2-piperidin-l-yl -propionic acid (/?)-(l-aza-bicyclo[2.2.2]oct-3- yl) ester (Example Ia) (120 mg) and 2-chloromethyl-benzooxazole (58 mg) in acetonitrile (2 mL) was heated at 80⁰C for 5 minutes. To the cooled mixture was added diethyl ether (10 mL) and the resulting precipitate collected by filtration, washed with diethyl ether and dried under vacuum at 60⁰C to afford the titled compound (150 mg).

m/e 474 [M]⁺ 1H NMR (400 MHz, DMSO) 57.90 (IH, d), 7.86 (IH, d), 7.59 - 7.48 (4H, m), 7.26 (2H, t), 7.20 - 7.14 (IH, m), 5.10 - 5.04 (IH, m), 5.03 - 4.94 (2H, m), 4.18 - 4.09 (IH₅ m), 3.78 - 3.67 (IH, m), 3.67 - 3.53 (2H, m), 3.52 - 3.46 (IH, m), 3.44 - 3.34 (IH, m), 2.45 - 2.28 (4H, m), 2.25 - 2.19 (IH, m), 2.01 - 1.78 (4H, m), 1.56 (3H, s), 1.54 - 1.45 (4H, m), 1.45 - 1.36 (2H, m). Example 8: (R)-l-(5-Methyl-benzooxazol-2-ylmethyl)-3-((S')-2-phenyl-2-piperidin-l- yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloromethy 1-5 -methyl-benzooxazole

A mixture of 2-amino-4-methyl~phenol (2 g) and 2-chloro-l,l,l-trimethoxy-ethane (3.01 g) in acetic acid (20 mL) was heated under reflux for 24h. The solvent was removed under reduced pressure and the residue treated with cone, sulfuric acid (5 mL). The brown solution was heated at 100⁰C for 2h, cooled , and quenched into water (200 mL). The mixture was made basic by the addition of solid potassium carbonate and the products extracted into diethyl ether (2 x 150 mL). The dried extracts were concentrated to an oil and the oil purified by chromatography on silica eluting with diethyl ether / wσhexane (3 : 7) to afford the titled compound as an oil (0.360 g).

¹H NMR (400 MHz, CDCl₃) δ 7.52 (IH, dd), 7.42 (IH, d), 7.20 (IH, dd), 4.74 (2H, d), 2.48 (3H, s).

Example 8: (R)-l-(5-Methyl-benzooxazol-2-ylmethyl)-3-((S)-2-phenyl-2-piperidin-l- yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

A mixture of (S^-phenyl^-piperidin-l-yl-propionic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3- yl) ester (Example Ia) (120 mg) and 2-chloromethyl-5-methyl-benzooxazole (Example 8a) (75 mg) in acetonitrile (2 mL) was heated under reflux for 5 minutes. The solution was cooled to ambient temperature and the solution diluted with diethyl ether (20 mL). The resulting solid was collected by filtration and dried to afford the titled compound as a solid (180 mg). m/e 488 [M]⁺

¹H NMR (400 MHz, DMSO) 57.72 (IH, d), 7.68 (IH, s), 7.54 - 7.49 (2H, m), 7.37 (IH, d), 7.29 - 7.22 (2H, m), 7.20 - 7.14 (IH, m), 5.10 - 5.04 (IH, m), 5.00 - 4.90 (2H, m), 4.16 - 4.06 (IH, m), 3.77 - 3.67 (IH, m), 3.65 - 3.51 (2H, m), 3.47 (IH, d), 3.43 - 3.34 (IH, m), 2.45 - 2.37 (2H, m), 2.36 - 2.28 (2H, m), 2.25 - 2.19 (IH, m), 2.01 - 1.77 (4H, m), 1.55 (3H, s), 1.54 - 1.45 (4H, m), 1.45 - 1.36 (2H, m), 2.48 (3H, s).

Example 9: (2?)-3-((S)-2-Phenyl-2-piperidm-l-yl-propionyloxy)-l-(l-phenyl-lH- pyrazol-4-ylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride

a) 4-Chloromethyl- 1 -phenyl- 1 H-pyrazole

(1 -Phenyl- lH-pyrazol-4-yl)~methanol (2 g) in dichloromethane (15 mL) was treated with thionyl chloride (10 mL) and the mixture stirred at 20⁰C for Ih. The resulting suspension was diluted with wøhexane (15 mL) and the solid collected by filtration to afford the subtitled compound as an off-white solid (1.120 g).

¹H NMR (400 MHz, CDCl₃) δ 8.61 (IH, s), 7.84 - 7.80 (3H, m), 7.53 - 7.47 (2H, m), 7.35 7.30 (IH, m), 4.77 (2H, s).

Example 9: (/?)-3-((5)-2-Phenyl-2-piperidin-l-yl-propionyloxy)-l-(l-pheπyl-liϊ- pyrazol-4-ylmethyl)- l-azonia-bicyclo[2.2.2]octane chloride

A mixture of (S)~2-phenyl-2-piperidin-l-yl-propionic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3- yl) ester (Example Ia) (100 mg) and 4-chloromethyl- 1 -phenyl- lH-pyrazole (Example 9a) (56 mg) were stirred at ambient temperature in acetonitrile (2 mL) for 17h. The solvent was removed under vacuum and the residue triturated with diethyl ether (20 mL). The resulting solid was collected by filtration and dried under vacuum at 50 ⁰C to afford the titled compound as a solid (150 mg).

m/e 499 [M]⁺

¹H NMR (400 MHz, DMSO) 5 8.62 (IH, s), 7.86 - 7.81 (3H, m), 7.57 - 7.51 (2H, m), 7.51 - 7.45 (2H, m), 7.40 - 7.34 (IH, m), 7.29 - 7.23 (2H, m), 7.18 - 7.12 (IH, m), 5.16 - 5.10 (IH, m), 4.42 (2H, d), 3.89 - 3.80 (IH, m), 3.55 - 3.45 (IH, m), 3.44 - 3.33 (2H, m), 3.21 - 3.09 (IH, m), 3.09 - 3.02 (IH, m), 2.47 - 2.39 (2H, m), 2.36 - 2.28 (2H, m), 2.27 - 2.21 (IH, m), 2.04 - 1.82 (4H, m), 1.54 (3H, s), 1.54 - 1.45 (4H, m), 1.44 - 1.36 (2H, m).

Example 10: (R)-3-((5')-2-Phenyl-2-piperidin-l-yl-propionyIoxy)-l-(l-phenyl-lH^r- [l^SJtriazoI^-ylmethy^-l-azonia-bicyclotZ.Z.Zloctane chloride a) 4-Chloromethyl-l-phenyl-lH-[l,2,3]triazole

(l-Phenyl-lH-[l,2,3]triazol-4-yl)-methanol (700 mg) was treated with thionyl chloride (2 mL) in dichloromethane (10 mL). The mixture was stirred and heated under reflux to form a solution. The excess reagent and solvent were removed under vacuum and the residue dissolved in dichloromethane (50 mL). The dichloromethane solution was passed through a pad of silica which was further washed with dichloromethane followed by diethyl ether. The eluants were combined and concentrated to give the sub-titled compound as a solid (600 mg).

¹H NMR (400 MHz, CDCl₃) 5 8.00 (IH, s), 7.73 - 7.68 (2H, m), 7.54 - 7.48 (2H, m), 7.47 - 7.41 (IH, m), 4.77 (2H, s).

Example 10: (Λ)-3-((S)-2-PhenyI-2-piperidin-l-yl-propionyloxy)-l-(l-phenyl-lH^r- [l^^triazoI-ΦylmethyO-l-azonia-bicyclop.l.ltøctane chloride

A mixture of (5)-2-phenyl-2-piperidin-l-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3- yl) ester (Example Ia) (200 mg) and 4-chloromethyl- 1 -phenyl- IH-[1, 2,3]triazole (Example 10a) (113 mg) in acetonitrile (2 mL) was heated under reflux for 15 minutes. To the cooled reaction mixture was added diethyl ether ( 1 mL) and the resulting solid collected by filtration to afford the titled compound as a colourless solid (200 mg).

m/e 500 [M]^{+ 1}H NMR (400 MHz, DMSO) δ 9.09 (IH, s), 7.99 - 7.93 (2H, m), 7.70 - 7.63 (2H, m), 7.59 - 7.47 (3H, m), 7.32 - 7.25 (2H, m), 7.17 (IH, t), 5.11 - 5.06 (IH, m), 4.75 - 4.65 (2H, m), 3.98 - 3.89 (IH, m), 3.62 - 3.51 (IH, m), 3.49 - 3.35 (2H, m), 3.30 - 3.17 (2H, m), 2.44 - 2.36 (2H, m), 2.35 - 2.27 (2H, m), 2.24 - 2.19 (IH, m), 1.99 - 1.77 (4H, m), 1.55 (3H, s), 1.53 - 1.46 (4H, m), 1.44 - 1.36 (2H, m).

Example 11: (i?)-l-Isopropyl-3-((S)-2-phenyl-2-piperidin-l-yI-propionyloxy)-l- azonia-bicyclo[2.2.2]octane iodide

A mixture of (5')-2-phenyl-2-piperidin-l-yl-propionic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3- yl) ester (Example Ia) (150 mg) and 2-iodo-propane (74 mg) in acetonitrile (2 mL) were stirred at ambient temperature for 5 days. Diethyl ether (20 mL) was added and the resulting solid collected by filtration to afford the titled compound as a solid (30 mg).

m/e 385 [M]⁺

¹H NMR (400 MHz, DMSO) δ 7.60 - 7.55 (2H, m), 7.37 (2H, t), 7.30 (IH, d), 5.14 - 5.08 (IH, m), 3.76 - 3.68 (IH, m), 3.52 - 3.37 (2H, m), 3.28 - 3.24 (IH, m), 2.94 - 2.83 (2H, m), 2.47 - 2.39 (2H, m), 2.38 - 2.29 (2H, m), 2.23 - 2.18 (IH, m), 1.97 - 1.74 (5H, m), 1.56 (3H, s), 1.56 - 1.49 (4H, m), 1.48 - 1.39 (2H, m), 1.21 - 1.14 (6H, m).

Example 12: (i?)-l-(5-PhenyI-[l,2,4]oxadiazol-3-yImethyl)-3-(2-piperidin-l-yI-2- thiophen-2-yl-propionyloxy)-l-azonia-bicydo[2.2.2]octane chloride (Isomer 1)

A mixture of 2-piperidin-l-yl-2-thiophen-2-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct- 3-yl) ester (Isomer 1) (Example 2b) (75 mg) and 3-chloromethyl-5-phenyl- [l,2,4]oxadiazole (40 mg) in acetonitrile (2 mL) was heated under reflux for 5 minutes. Diethyl ether (3 mL) was added and the resulting solid collected by filtration. Recrystallisation from acetone (2 mL) and drying under vacuum afforded the titled compound as a solid (86 mg).

m/e 507 [M]⁺

¹H NMR (400 MHz, DMSO) δ 8.19 - 8.13 (2H, m), 7.82 - 7.76 (IH, m), 7.73 - 7.66 (2H, m), 7.38 (IH, dd), 7.05 - 7.03 (IH, m), 6.84 (IH, dd), 5.14 - 5.08 (IH, m), 4.88 (2H, dd), 4.15 - 4.07 (IH, m), 3.78 - 3.68 (IH, m), 3.68 - 3.51 (2H, m), 3.48 - 3.36 (2H, m), 2.48 - 2.40 (2H, m), 2.38 - 2.30 (2H, m), 2.31 - 2.26 (IH, m), 2.05 - 1.83 (4H, m), 1.60 (3H, s), 1.53 - 1.44 (4H, m), 1.44 - 1.36 (2H, m).

Example 13: (Λ)-l-(3-Phenyl-isoxazol-5-yImethyl)-3-(2-piperidin-l-yl-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane chloride (Isomer 1)

A mixture of 2-piperidin-l-yl-2-thiophen~2-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct- 3-yl) ester (Isomer 1) (Example 2b) (77 mg) and S-chloromethyl-S-phenyl-isoxazole (45 mg) in acetonitrile (2 mL) was stirred at room temperature for 72h. Diethyl ether (10 mL) was added and the mixture stirred for 48h. The resulting off-white solid was collected by filtration, washed with diethyl ether and dried under vacuum at room temperature. The solid was recrystallised from acetone and the solid collected by filtration. The solid was washed with acetone and dried under vacuum at room temperature to afford the titled compound as a white solid (51 mg).

m/e 506 [M]⁺

¹H NMR (400 MHz, DMSO) 67.97 - 7.91 (2H, m), 7.61 - 7.54 (3H, m), 7.48 (IH, s), 7.34 (IH, dd), 7.04 (IH, dd), 6.83 (IH, dd), 5.18 - 5.11 (IH, m), 4.88 (2H, dd), 3.99 (IH, ddd), 3.65 (IH, t), 3.59 - 3.41 (2H, m), 3.39 - 3.28 (IH, m), 3.25 (IH, d), 2.47 - 2.38 (2H, m), 2.37 - 2.25 (3H, m), 2.06 - 1.85 (4H, m), 1.59 (3H, s), 1.53 - 1.43 (4H, m), 1.43 - 1.34 (2H, m).

Example 14: (/?)-l-[3-(4-Methoxy-phenyl)-[l,2,4]oxadiazol-5-ylmethyl]-3-(2- piperidin-l-yl-2-thiophen-2-yI-propionyloxy)-l-azonia-bicyclo[2.2.2]octane chloride (Isomer 1)

A mixture of 2-piperidin-l-yl-2-thiophen-2-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct- 3-yl) ester (Isomer 1) (Example 2b) (78 mg) and 5-chloromethyl-3-(4-methoxy-phenyl)- [l,2,4]oxadiazole (61 mg) was stirred at room temperature over 72h. The resulting precipitate was collected by filtration, washed with acetonitrile and dried at room temperature under vacuum to give a solid. Recrystallisation from acetonitrile gave crystals that were collected by filtration, washed with acetonitrile and dried under vacuum at room temperature to afford the titled compound as white crystals (75 mg).

m/e 537 [M]⁺

¹H NMR (400 MHz, DMSO) δ 8.02 - 7.97 (2H, m), 7.40 (IH, dd), 7.19 - 7.13 (2H, m), 7.06 (IH, dd), 6.87 (IH, dd), 5.15 - 5.04 (3H, m), 4.13 (IH, ddd), 3.86 (3H, s), 3.76 (IH, t), 3.71 - 3.42 (4H, m), 2.48 - 2.40 (2H, m), 2.39 - 2.25 (3H, m), 2.06 - 1.84 (4H, m), 1.61 (3H, s), 1.54 - 1.44 (4H, m), 1.44 - 1.35 (2H, m).

Example 15: (Z2)-l-[3-(5-Methyl-pyridin-3-yI)-propyI]-3-(2-piperidin-l-yl-2-thiophen- 2-yl-propionyloxy)-l-azonia-bicycIo[2.2.2]octane bromide (Isomer 1)

a) 3-(5-Methyl-pyridin-3-yl)-ρrop-2-yn-l-ol

B2008/002072

50

Ethylene glycol dimethyl ether (50 mL) and water (50 mL) were added to a mixture of 3- bromo-5-methyl-pyridine (5.03 g), triphenylphosphine (0.622 g), copρer(I) iodide (0.045 mL), potassium carbonate (4.13 mL) and 10% palladium on carbon (0.670 g), and the resulting mixture was stirred at room temperature for 30 minutes. Propargyl alcohol (4.32 mL) was then added and the mixture was heated to 80⁰C and stirred for 48h. The mixture was allowed to cool and was then concentrated under reduced pressure to remove most of the ethylene glycol dimethyl ether. The remaining slurry was acidified with 2M hydrochloric acid and filtered through Celite. The Celite pad was further washed with 2M hydrochloric acid and toluene. The filtrate was extracted with toluene and the organic phase was discarded. The aqueous phase was basified with 10% aqueous sodium hydroxide solution and extracted three times with ethyl acetate. The combined organic extracts were dried (MgSO₄), filtered and purified by flash chromatography on silica eluted with methanol / dichloromethane (1 : 9) to afford the titled compound as a pale yellow solid (1.45 g).

¹H NMR (400 MHz, CDCl₃) δ 8.58 (IH, d), 8.35 (IH, d), 7.55 (IH, s), 4.50 (2H, s), 3.55 (lH, br s), 2.32 (3H, s).

b) 3-(5-Methyl-pyridin-3-yl)-propan- l-ol

10% Palladium on carbon (0.10Og) was added to a solution of 3-(5-methyl-pyridin-3-yl)- prop-2-yn-l-ol (Example 15a) (1.457 g) in ethanol (20 mL). The resulting suspension was stirred under 1 atmosphere of hydrogen for 5h. The catalyst was removed by filtration through Celite and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography on silica eluted with methanol / dichloromethane (5 : 95) to afford the product as a pale yellow oil (1.368 g). ¹H NMR (400 MHz, CDCl₃) δ 8.27 (2H, s), 7.33 (IH, s), 3.68 (2H, t), 2.69 (2H, t), 2.31 (3H, s), 1.89 (2H, tt).

c) 3-(3-Bromo-propyl)-5-methyl-pyridine hydrob^'romide

A solution of 3-(5-methyl-pyridin-3-yl)-propan-l-ol (Example 15b) (1.36 g) in 48% Hydrobromic acid (6.5 mL) was heated at reflux overnight. The cooled solution was concentrated under reduced pressure and the residue was dissolved in wøpropanol and re- concentrated. This process was repeated three more times. The residue was dissolved in isopropanol, decolourised by boiling with activated charcoal, and filtered. The clear solution was concentrated under vacuum and the residue dissolved in acetone and left to crystallise in the freezer for 48h. The crystals were removed by filtration, washed with acetone and dried under vacuum, at room temperature overnight to give the sub-titled compound as an off-white solid (0.869 g).

¹H NMR (400 MHz, CDCl₃) 58.49 (IH, s), 8.46 (IH, s), 8.35 (IH, s), 3.49 (2H, t), 3.00 (2H, t), 2.51 (3H, s), 2.24 (2H, quint.).

Example 15: (i?)-l-[3-(5-Methyl-pyridin-3-yl)-propyl]-3-(2-piperidin-l-yl-2-thiophen- 2-yl-propionyloxy)-l-azonia-bicycIo[2.2.2]octane bromide (Isomer 1)

3-(3-Bromo-propyl)-5-methyl-pyridine hydrobromide (Example 15c) (102 mg) was dissolved in water (2 mL), basified with 10% aqueous sodium hydroxide solution (2 mL) s and extracted twice with diethyl ether. The combined organic phases were washed with brine, dried (MgSO₄) and concentrated under vacuum to give 3-(3-bromopropyl)-5- methylpyridine free base as a yellow oil (0.072 g). 2-Piperidin-l-yl-2-thiophen-2~yl- propionic acid (R)-(I -aza-bicyclo[2.2.2]oct-3-yl) ester (Isomer 1) (Example 2b) (117 mg) was added to the residue together with acetonitrile (3 mL), and the resulting mixture waso stirred at room temperature for 72h. The solution was diluted with diethyl ether (10 mL) to give a cloudy solution that crystallised after stirring for 15 minutes. The solid was removed by filtration, washed with diethyl ether and dried under vacuum at room temperature. The solid was recrystallised from acetone, the crystals were collected by filtration, washed with acetone and dried under vacuum at room temperature to afford thes titled product as white crystals (46 mg).

m/e 482 [M]⁺

¹H NMR (400 MHz, DMSO) δ 8.28 (2H, dd), 7.51 - 7.48 (IH, m), 7.46 (IH, dd), 7.08 (IH, dd), 6.92 (IH, dd), 5.12 - 5.06 (IH, m), 3.84 (IH, ddd), 3.54 - 3.44 (IH, m), 3.43 - 3.260 (2H, m), 3.26 - 3.13 (3H, m), 3.07 (IH, d), 2.59 - 2.41 (4H, m), 2.39 - 2.31 (2H, m), 2.29 (3H, s), 2.27 - 2.21 (IH, m), 2.00 - 1.78 (6H, m), 1.60 (3H, s), 1.55 - 1.46 (4H, m), 1.46 - 1.37 (2H, m). Example 16: (i?)-l-(5-PhenyI-isoxazol-3-ylmethyl)-3-(2-piperidin-l-yl-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide (Isomer 1)

2-Piperidin-l-yl-2~thiophen-2-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Isomer 1) (Example 2b) (100 mg) in acetonitrile (2 mL) was treated with 3-chloromethyl- 5-phenyl-isoxazole (67 mg) and the mixture heated to 80⁰C for 5 minutes. Diethyl ether (5 mL) was added to the cooled solution and the titled compound was collected by filtration (98 mg).

m/e 506 [M]⁺

¹H NMR (400 MHz, DMSO) δ 7.97 - 7.90 (2H, m), 7.65 - 7.55 (3H, m), 7.32 (IH, dd), 7.29 (IH, s), 7.03 (IH, dd), 6.82 (IH, dd), 5.18 - 5.12 (IH, m), 4.74 (2H, dd), 4.05 - 3.96 (IH, m), 3.74 - 3.62 (IH, m), 3.61 - 3.44 (2H, m), 3.37 - 3.21 (2H, m), 2.47 - 2.38 (2H, m), 2.36 - 2.27 (3H, m), 2.06 - 1.86 (4H, m), 1.58 (3H, s), 1.52 - 1.44 (4H, m), 1.43 - 1.35 (2H, m).

Example 17: (7?)-l-{2-[4-(2-Hydroxy-ethyl)-phenyl]-ethyl}-3-((S)-2-phenyI-2- piperidin-l-yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-[4-(2-Bromo-ethyl)-phenyl]-ethanol

2-[4-(2-Hydroxy-ethyl)-phenyl]-ethanol (2.509 g) was suspended in toluene (50 mL) and 48% aq HBr (2.563 g, 1.72 mL) was added. The mixture was heated at reflux for 30h and then the mixture allowed to cool to room temperature. The solvent was evaporated under reduced pressure and the residue was purified by chromatography on silica eluting with istfhexane : diethyl ether (7 / 1 to 1 / 1) to afford the titled compound (2.309 g).

¹H NMR (400 MHz, DMSO) 67.14 - 7.21 (4H, m), 3.86 (2H, q), 3.56 (2H, t), 3.14 (2H, t), 2.86 (2H, t), 1.35 (IH, t).

Example 17: (R)-l-{2-[4-(2-Hydroxy-ethyl)-phenyl]-ethyl}-3-((S)-2-phenyl-2- piperidin-l-yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

To a solution of (5)-2-phenyl-2-piperidin-l-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct- 3-yl) ester (Example Ia) (106 mg) in acetonitrile (2 mL) was added 2-[4-(2-bromo-ethyl)- phenyl]-ethanol (Example 17a) (85 mg) and the reaction stirred at room temperature over 72 h. The mixture was heated to 40⁰C for 2 h and then allowed to cool to room temperature. The mixture was partially evaporated to approximately half its initial volume and diethyl ether added. The resulting oil was washed with further aliquots of diethyl ether which were removed by decantation. The residue was left to stand overnight in diethyl ether and the resulting solid collected by filtration and dried under vacuum at 60⁰C to afford the titled compound as a white solid (90 mg).

m/e 491 [M]⁺ 1H NMR (400 MHz, DMSO) δ 7.54 (2H, d), 7.36 (2H, t), 7.31 - 7.23 (IH, m), 7.18 (4H, s), 5.17 - 5.09 (IH, m), 3.95 - 3.86 (IH, m), 3.61 (2H, t), 3.58 - 3.51 (IH, m), 3.51 - 3.36 (3H, m), 3.28 - 3.18 (IH, m), 3.14 (IH, d), 3.08 - 2.88 (4H, m), 2.71 (2H, t), 2.49 - 2.32 (4H, m), 2.27 - 2.20 (IH, m), 2.05 - 1.80 (4H, m), 1.59 (3H, s), 1.58 - 1.49 (4H, m), 1.50 - 1.40 (2H, m).

Example 18: (R)-l-[2-(lH-Indol-3-yl)-ethyl]-3-((5)-2-phenyl-2-piperidin-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

To a solution of GS)-2-phenyl-2-piperidin-l-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct- 3-yl) ester (Example Ia) (175 mg) in acetonitrile (2 mL) was added 3-(2-bromo-ethyl)-lH- indole (137 mg) and the mixture stirred at room temperature for 18h. The mixture was partially evaporated to approximately half its initial volume and diethyl ether added. The resulting oil was washed with further aliquots of diethyl ether which were removed by decantation. The residue was left to stand for 2h in diethyl ether and the resulting solid collected by filtration and dried under vacuum at 60⁰C to afford the titled compound as a white solid (119 mg). m/e 486 [M]⁺

¹H NMR (400 MHz, DMSO) δ 7.60 (2H, d), 9.33 (IH, s), 7.46 (IH, d), 7.35 - 7.29 (3H, m), 7.23 (IH, t), 7.15 (IH, t), 7.07 (IH, t), 7.01 (IH, d), 5.00 (IH, t), 3.86 - 3.77 (2H, m), 3.72 - 3.62 (IH, m), 3.56 - 3.46 (IH, m), 3.43 - 3.33 (IH, m), 3.31 - 3.21 (IH, m), 2.75 - 2.61 (2H, m), 2.57 - 2.45 (4H, m), 2.36 - 2.24 (3H, m), 1.99 - 1.77 (4H, m), 1.65 - 1.55 (7H, m), 1.53 - 1.46 (2H, m).

Example 19: (R)-l-(5-Phenyl-oxazol-2-ylmethyI)-3-((S)-2-phenyl-2-piperidin-l-yI- propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-Chloromethyl-5-phenyl-oxazole

2-Amino-l-phenyl-ethanone hydrochloride (1.4 g) in acetic acid (20 mL) was treated with 2-chloro- 1,1,1 -trimethoxy-ethane ( 1.9 g) and the mixture stirred and heated under reflux for 2Oh. The cooled reaction mixture was poured into satd aq. sodium bicarbonate (150 mL) and the products extracted into diethyl ether (200 mL). The combined organics was dried (MgSO₄) and concentrated to dryness. The residue was purified by chromatography on silica eluting with diethyl ether / isohexane (3 / 7) to afford the sub-titled compound as an off-white solid (0.090 g).

¹H NMR (400 MHz, CDCl₃) δ 7.68 - 7.64 (2H, m), 7.47 - 7.40 (2H, m), 7.38 - 7.33 (IH, m), 7.32 (IH, s), 4.68 (2H, s).

Example 19: (2?)-l-(5-Phenyl-oxazol-2-ylmethyl)-3-((S)-2-phenyl-2-piperidin-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

To a solution of (S)-2-phenyl-2-piperidin-l-yl -propionic acid (/?)-( l-aza-bicyclo[2.2.2]oct- 3-yl) ester (Example Ia) (118 mg) in acetonitrile (2 mL) was stirred and heated at 80⁰C with 2-chloromethyl-5-phenyl-oxazole (Example 19a) (80 mg) for 10 minutes. The solution was cooled to room temperature and diethyl ether (5 mL) was added. The resulting gum was triturated with diethyl ether to afford a solid which was recrystallised from acetone / diethyl ether to afford the titled compound as a colourless solid (80 mg).

m/e 500 [M]⁺

¹H NMR (400 MHz, DMSO) 5 7.90 (IH, s), 7.80 - 7.74 (2H, m), 7.58 - 7.42 (5H, m), 7.34 - 7.26 (2H, m), 7.23 - 7.16 (IH, m), 5.12 - 5.04 (IH, m), 4.82 (2H, dd), 4.08 - 3.99 (IH, m), 3.69 - 3.45 (3H, m), 3.43 - 3.25 (2H, m), 2.45 - 2.28 (4H, m), 2.26 - 2.18 (IH, m), 2.03 - 1.76 (4H, m), 1.55 (3H, s), 1.53 - 1.44 (4H, m), 1.44 - 1.36 (2H, m).

Example 20: (R)-l-(5-PhenyI-isoxazol-3-ylmethyI)-3-((S)-2-phenyl-2-piperidin-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

(S)-2-Phenyl-2-piperidin-l-yl-proρionic acid (R)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (103 mg) and S-chloromethyl-S-phenyl-isoxazole (70 mg) were dissolved in acetonitrile (3 mL) and stirred at room temperature for 18h. The resulting solid was collected by filtration to afford the titled compound (57 mg).

m/e 500 [M]⁺

¹H NMR (400 MHz, DMSO) δ 7.94 (2H, dd), 7.64 - 7.55 (3H, m), 7.51 (2H, d), 7.31 - 7.24 (3H, m), 7.14 (IH, t), 5.13 - 5.08 (IH, m), 4.72 (2H, dd), 4.04 - 3.95 (IH, m), 3.67 - 3.58 (IH, m), 3.57 - 3.42 (2H, m), 3.30 - 3.21 (2H, m), 2.44 - 2.35 (2H, m), 2.35 - 2.26 (2H, m), 2.26 - 2.21 (IH, m), 2.04 - 1.79 (4H, m), 1.54 (3H, s), 1.52 - 1.44 (4H, m), 1.43 - 1.35 (2H, m).

Example 21 : (2?)-l-(3-Phenyl-isoxazol-5-ylmethyl)-3-((S)-2-phenyl-2-piperidin-l-yI- propionyloxy)-l-azonia-bicyclo[2.2.2]octane choride

(S)-2-Phenyl-2-piperidin-l-yl-propionic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (55 mg) in acetonitrile (0.5 mL) was treated with 5-chloromethyl-3-ρhenyl- isoxazole (37 mg) and the mixture was stirred at room temperature for 18h. Diethyl ether was added and the resulting white solid was filtered off, washed with diethyl ether and dried to afford the titled compound (70 mg).

m/e 500 [M]^{+ 1}H NMR (400 MHz, DMSO) δ 7.96 - 7.92 (2H, m), 7.60 - 7.55 (3H, m), 7.51 (2H, d), 7.47 (IH, s), 7.29 (2H, t), 7.17 (IH, t), 5.13 - 5.08 (IH, m), 4.86 (2H, dd), 4.02 - 3.94 (IH, m), 3.65 - 3.55 (IH, m), 3.55 - 3.39 (2H, m), 3.34 - 3.24 (2H, m,), 2.44 - 2.36 (2H, m), 2.36 - 2.28 (2H, m), 2.23 (IH, s), 2.02 - 1.78 (4H, m), 1.55 (3H, s), 1.53 - 1.45 (4H, m), 1.43 - 1.36 (2H, m).

Example 22: (R)-l-(3-Phenyl-[l,2,4]oxadiazoI-5-ylmethyl)-3-((S)-2-phenyl-2- piperidin-l-yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane choride

(S)-2-Phenyl-2-piperidin-l-yl-propionic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (65 mg) in acetonitrile (0.5 mL) was treated with 5-chloromethyl-3-phenyl- [l,2,4]oxadiazole (44 mg) and the mixture was stirred at room temperature for 18h. Diethyl ether was added and the resulting white solid was filtered off and washed with diethyl ether to afford the titled compound (75 mg).

m/e 501 [M]⁺

¹H NMR (400 MHz, DMSO) δ 8.08 - 8.05 (2H, m), 7.69 - 7.60 (3H, m), 7.53 (2H, d), 7.31 (2H, t), 7.22 (IH, t), 5.17 - 5.07 (3H, m), 4.18 - 4.10 (IH, m), 3.78 - 3.42 (5H, m), 2.46 - 2.38 (2H, m), 2.38 - 2.30 (2H, m), 2.23 (IH, s), 2.03 - 1.76 (4H, m), 1.57 (3H, s), 1.54 - 1.46 (4H, m), 1.45 - 1.37 (2H, m).

Example 23: (2?)-l-(5-PhenyI-[l,2,4]oxadiazol-3-ylmethyl)-3-((S)-2-phenyl-2- piperidin-l-yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane choride

(5)-2-Phenyl-2-piperidin-l-yl -propionic acid (/?)-( l-aza~bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (65 mg) in acetonitrile (0.5 mL) was treated with 3-chloromethyl-5-phenyl- [l,2,4]oxadiazole (41 mg) and the mixture was stirred at room temperature for 36h. Diethyl ether was added and the resulting white solid was filtered off and washed with diethyl ether to afford the titled compound (80 mg).

m/e 501 [M]⁺ 1H NMR (400 MHz, DMSO) δ 8.18 - 8.14 (2H, m), 7.79 (IH, t), 7.70 (2H, t), 7.52 (2H, d), 7.29 (2H, t), 7.19 (IH, t), 5.10 - 5.05 (IH, m), 4.86 (2H, dd), 4.13 - 4.05 (IH, m), 3.74 - 3.63 (IH, m), 3.63 - 3.50 (2H, m), 3.46 (IH, d), 3.42 - 3.33 (IH, m), 2.46 - 2.37 (2H, m), 2.37 - 2.29 (2H, m), 2.23 (IH, s), 2.02 - 1.76 (4H, m), 1.55 (3H, s), 1.53 - 1.46 (4H, m), 1.44 - 1.37 (2H, m).

Example 24: (_JR)-l-(2-Phenyl-oxazol-4-yImethyl)-3-((S)-2-phenyI-2-piperidin-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane choride

a) 4-Chloromethyl-2-phenyl-oxazole

Benzamide (200 mg) and l,3-dichloro-propan-2-one (210 mg) in ethanol (3 mL) were heated at 80°C for 8 h. The mixture was allowed to cool and the resulting solid filtered off and washed with ethanol. The compound was purified by chromatography eluting with dichloromethane to yield a the sub-titled compound as a colourless oil which solidified (165 mg).

¹H NMR (400MHz, CDCl₃) 6 8.07 - 8.03 (m, 2H), 7.71 (s, IH), 7.50 - 7.41 (m, 3H), 4.58 (s, 2H).

Example 24: (_JR)-l-(2-Phenyl-oxazol-4-ylmethyI)-3-((S)-2-phenyI-2-piperidin-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane choride

(■S)-2-Phenyl-2-piperidin-l-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (70 mg) in acetonitrile (0.5 mL) was treated with 4-chloromethyl-2-phenyl- oxazole (44 mg) (Example 24a) and the mixture was stirred at room temperature for 72h. The resulting white solid was filtered off and washed with diethyl ether to afford the titled compound (52 mg).

m/e 500 [M]⁺

¹H NMR (400 MHz, DMSO) δ 8.45 (IH, s), 8.03 - 7.98 (2H, m), 7.62 - 7.58 (3H, m), 7.48 (2H, d), 7.23 (2H, t), 7.13 (IH, t), 5.11 - 5.06 (IH, m), 4.50 (2H, dd), 3.98 - 3.90 (IH, m), 3.62 - 3.40 (3H, m), 3.28 - 3.09 (2H, m), 2.42 - 2.34 (2H, m), 2.34 - 2.26 (2H, m), 2.23 (IH, s), 2.02 - 1.78 (4H, m), 1.52 (3H, s), 1.50 - 1.34 (6H, m). Example 25: (R)-l-[3-(3-Fluoro-phenyl)-[l,2,4]oxadiazol-5-ylmethyl]-3-((5)-2-phenyl- 2-piperidin-l-yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane choride

a) 5-Chloromethyl-3-(3-fluoro-phenyl)-[l ,2,4]oxadiazole

3-Fluoro-iV-hydroxy-benzamidine (0.62 g) in acetone (2 mL) was treated with potassium carbonate (0.556 g) and the mixture was cooled in ice. Chloroacetyl chloride (0.322 mL) in acetone (2 mL) was added gradually and then the mixture was stirred for 2h at room temperature. The solvent was evaporated and the resulting white solid was triturated with water and then filtered and washed with more water. The solid was heated at 135⁰C in xylene (3 mL) for Ih and then concentrated under vacuum. The resulting yellow oil was purified by chromatography on silica eluting with dichloromethane to yield the sub-titled compound as a colourless oil (0.850 g).

¹H NMR (400 MHz, CDCl₃) δ 7.89 (IH, d), 7.79 (IH, ddd), 7.47 (IH, td), 7.23 (IH, tdd), 4.75 (2H, s).

Example 25: (R)-l-[3-(3-Fluoro-phenyI)-[l,2,4]oxadiazol-5-ylmethyl]-3-((5)-2-pheπyl- 2-piperidin-l-yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane choride

(S)-2-Phenyl-2-piperidin-l-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (70 mg) in acetonitrile (0.5 mL) was treated with 5-chloromethyl-3-(3- fluoro-phenyl)-[l,2,4]oxadiazole (48 mg) and the mixture was stirred at room temperature overnight. Diethyl ether was added and the resulting white solid was collected by filtration. The solid was washed with diethyl ether and dried under vacuum to afford the titled compound (92 mg).

m/e 519 [M]⁺

¹H NMR (400 MHz, DMSO) δ 7.92 (IH, d), 7.82 (IH, ddt), 7.70 (IH, td), 7.57 - 7.51 (3H, m), 7.32 (2H, t), 7.22 (IH, t), 5.21 - 5.07 (3H, m), 4.19 - 4.11 (IH, m), 3.79 - 3.44 (5H, m), 2.46 - 2.38 (2H, m), 2.38 - 2.30 (2H, m), 2.23 (IH, s), 2.03 - 1.74 (4H, m), 1.57 (3H, s), 1.54 - 1.36 (6H, m).

Example 26: (i?)-l-(2-Phenyl-oxazol-5-yImethyl)-3-((S)-2-phenyl-2-piperidin-l-yI- propionyloxy)-l-azonia-bicyclo[2.2.2]octane choride

(S)-2-Phenyl-2-piperidin-l-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester

(Example Ia) (70 mg) in acetonitrile (0.5 mL) was treated with 5-bromomethyl-2-phenyl- oxazole (prepared as described in WO05/009941) (110 mg) and the mixture was stirred at room temperature for 18h. Diethyl ether was added and the resulting white solid was filtered off, washed with diethyl ether and dried under vacuum to afford the titled compound (48 mg). m/e 500 [M]⁺

¹H NMR (400 MHz, DMSO) 8.05 - 7.99 (2H, m), 7.63 - 7.58 (4H, m), 7.50 (2H, d), 7.28 (2H, t), 7.18 (IH, t), 5.12 - 5.07 (IH, m), 4.76 (2H, dd), 4.00 - 3.91 (IH, m), 3.62 - 3.37 (3H, m), 3.31 - 3.19 (2H, m), 2.44 - 2.36 (2H, m), 2.36 - 2.27 (2H, m), 2.22 (IH, s), 2.02 - 1.76 (4H, m), 1.54 (3H, s), 1.53 - 1.44 (4H, m), 1.44 - 1.35 (2H, m).

Example 27: (l?)-3-((S)-2-Phenyl-2-piperidin-l-yl-propionyloxy)-l-(3-pyridin-4-yl- [l,2,4]oxadiazoI-5-ylmethyl)-l-azonia-bicyclo[2.2.2]octane choride

a) 4-(5-Chloromethyl-[ 1 ,2,4]oxadiazol-3-yl)-pyridine

iV-Hydroxy-isonicotinamidine (200 mg) in chloroform (2.8 mL) was treated with chloroacetyl chloride (0.117 mL) and the mixture was cooled in ice. Triethylamine (0.224 mL) was added gradually and the mixture was stirred at room temperature for 18h. It was extracted (x3) with water and then the organic layer was filtered and the solid was washed with ethanol. The solid was heated in xylene (2.5 mL) at 140⁰C for Ih. The solution was concentrated to an oil and purified by chromatography on silica eluting with ethyl acetate / wøhexane (1 : 1) to afford the titled compound as a solid (38 mg).

¹H NMR (400 MHz, CDCl₃) 6 8.81 (2H, dd), 7.95 (2H, dd), 4.78 (2H, s).

Example 27: (2?)-3-((S)-2-Phenyl-2-piperidin-l-yl-propionyloxy)-l-(3-pyridin-4-yl- [l,2,4]oxadiazol-5-ylmethyl)-l-azonia-bicyclo[2.2.2]octane choride

(5)-2-Phenyl-2-piperidin-l-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (55 mg) in acetonitrile (0.5 mL) was treated with 4-(5-chloromethyl- [l,2,4]oxadiazol-3-yl)-pyridine (Example 27a) (38 mg) and the mixture was stirred at room temperature for 18h. Diethyl ether was added and the resulting white solid was filtered off, washed with diethyl ether and dried under vacuum to afford the titled compound (50 mg).

m/e 502 [M]⁺

¹H NMR (400 MHz, DMSO) δ 8.88 (2H, dd), 7.99 (2H, dd), 7.53 (2H, d), 7.32 (2H, t), 7.23 (IH, t), 5.21 - 5.12 (2H, m), 5.11 - 5.06 (IH, m), 4.19 - 4.10 (IH, m), 3.78 - 3.44 (5H, m), 2.46 - 2.38 (2H, m), 2.38 - 2.30 (2H, m), 2.23 (IH, s), 2.03 - 1.75 (4H, m), 1.57 (3H, s), 1.54 - 1.37 (6H, m).

Example 28: (_JR)-l-{2-[2-(4-ChIoro-phenyl)-thiazoI-4-yl]-ethyl}-3-((5)-2-phenyl-2- piperidin-l-yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane choride

B2008/002072

66

(S)~2-Phenyl-2-piperidin-l-yl-propionic acid (/?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (75 mg) in acetonitrile (0.5 mL) was treated with 4-(2-chloro-ethyl)-2-(4- chloro-phenyl)-thiazole (prepared as described in DE1913471) (68 mg) and the mixture was stirred at room temperature for 10 days. Diethyl ether was added and the resulting solid was collected by filtration, washed with diethyl ether and dried to afford the titled compound (60 mg).

m/e 564 [M]⁺

¹H NMR (400 MHz, DMSO) 67.96 (2H, dt), 7.61 - 7.54 (5H, m), 7.37 (2H, t), 7.28 (IH, t), 5.12 - 5.07 (IH, m), 3.98 - 3.90 (IH, m), 3.67 - 3.54 (3H, m), 3.53 - 3.37 (2H, m), 3.28 - 3.17 (4H, m), 2.47 - 2.31 (4H, m), 2.21 (IH, s), 2.04 - 1.71 (4H, m), 1.57 (3H, s), 1.56 - 1.48 (4H, m), 1.47 - 1.39 (2H, m).

Example 29: (/?)-l-(3-Benzo[ϋ»]thiophen-2-yl-propyl)-3-((S)-2-phenyl-2-piperidin-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 3-Benzo[b]thiophen-2-yl-propan- 1 -ol

Lithium aluminium hydride (IM solution in THF, 5.82 mL) was added to THF (10 mL) in a dried, nitrogen filled 3-necked flask. Then 3-benzo[έ>]thiophen-2-yl-propionic acid (1 g) was added in portions and the mixture was stirred for 18h. Ethyl acetate in wohexane (2-3 mL) was added dropwise. Then the mixture was poured gradually onto dilute HCl. It was extracted into ethyl acetate (x3) and the organic was washed with water, dried and the solvent was evaporated to yield the sub-titled compound as an off white solid (1.34 g).

¹H NMR (400 MHz, DMSO) δ 7.86 (IH, d), 7.72 (IH, d), 7.35 - 7.23 (2H, m), 7.15 (IH, d), 3.48 (2H, t), 2.93 (2H₅1), 1.87 - 1.80 (2H, m).

b) 2-(3-Bromo-propyl)-benzo[b]thiophene

3-Benzo[b]thiophen-2-yl-propan-l-ol (Example 29a) (0.4 g) in hydrobromic acid (3 niL) was heated at 105⁰C for 5h. The reaction mixture was evaporated and azeotroped with isopropanol (x3) to yield the sub-titled compound as an off-white solid (0.414 g).

¹H NMR (400 MHz, DMSO) δ 7.88 (IH, dd), 7.74 (IH, d), 7.31 (2H, dtd), 7.21 (IH, s), 3.59 (2H, t), 3.04 (2H, t), 2.22 (2H, quint.).

Example 29: (/?)-l-(3-Benzo[Z>]thiophen-2-yl-propyl)-3-((S)-2-phenyl-2-piperidin-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

(S)-2-Phenyl-2-piperidin-l-yl-propionic acid (/?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (100 mg) in acetonitrile (0.5 mL) was treated with 2-(3-bromo-propyl)- benzo[b]thiophene (Example 29b) (97 mg) and the mixture was stirred at room temperature for 18h. The solvent was evaporated under reduced pressure and diethyl ether was added. The solvents were decanted, acetone and diethyl ether were added and the product was triturated. The resulting white solid was filtered off, washed with diethyl ether and dried to afford the titled compound (142 mg).

m/e 517 [M]^{+ 1}H NMR (400 MHz, DMSO) δ 7.92 (IH, d), 7.77 (IH, d), 7.54 (2H, d), 7.38 - 7.28 (4H, m), 7.25 - 7.19 (2H, m), 5.09 - 5.03 (IH, m), 3.87 - 3.79 (IH, m), 3.51 - 3.16 (5H, m), 3.14 - 3.01 (2H, m), 2.92 (2H, t), 2.51 - 2.38 (2H, m), 2.38 - 2.29 (2H, m), 2.19 (IH, s), 2.09 - 1.82 (4H, m), 1.81 - 1.72 (2H, m), 1.55 (3H, s), 1.59 - 1.47 (4H, m), 1.46 - 1.37 (2H, m).

Example 30: (_JR)-3-((5)-2-Phenyl-2-piperidin-l-yI-propionyIoxy)-l-(3-quinoxalm-6-yl- propyl)-l-azonia-bicyclo[2.2.2]octane bromide

a) 3-Quinoxalin-6-yl-prop-2-yn-l-ol

To 6-bromo-quinoxaline (1 g) in acetonitrile (25 mL) was added propargyl alcohol (0.285 mL) and triethylamine (1.334 mL) followed by copper(I) iodide (10 mg) and bis(triphenylphosphine)palladium chloride (34 mg). The reaction mixture was stirred under nitrogen and then heated at 80⁰C for 18h. The mixture was allowed to cool and then filtered. The filtrate was evaporated and then purified by chromatography on silica eluting with ethylacetate / isohexane (1 : 1 to 1 : 0) to afford the sub-titled compound as a solid (0.7 g).

¹H NMR (400 MHz, DMSO) δ 8.97 (2H, dd), 8.13 (IH, d), 8.10 (IH, d), 7.84 (IH, dd), 5.45 (IH, t), 4.39 (2H, d).

b) 3-Quinoxalin-6-yl-propan-l-ol

3-Quinoxalin-6-yl-prop-2-yn-l-ol (Example 30a) (1.731 g) in ethanol (30 mL) was treated with 10% palladium on carbon (0.02 g) in ethanol and then hydrogen at 3 bar for 2h with rapid stirring. The reaction mixture was filtered and the catalyst was washed with ethanol. The filtrate was evaporated to yield dark brown oil which was redissolved in ethanol and further 10% palladium on carbon catalyst was added. The reaction mixture was rapidly stirred under hydrogen at 3.5 bar for 5h. The catalyst was filtered off, washed with ethanol and the filtrate concentrated under reduced pressure. The resulting viscous brown oil was purified by chromatography eluting with ethyl acetate as eluent to yield an oil. Repetition of this purification afforded the sub-titled compound as a oil (0.12 g).

¹H NMR (400 MHz, DMSO) δ 8.90 (2H, dd), 8.02 (IH, d), 7.89 (IH, d), 7.75 (IH, dd), 4.54 (IH, t), 3.46 (2H, dd), 2.89 (2H, t), 1.85 (2H, dt).

c) 6-(3-Bromo-propyl)-quinoxaline hydrobromide salt

3-Quinoxalin-6-yl-propan-l-ol (Example 30b) (120 mg) in hydrobromic acid (3 mL) was heated at 105⁰C for 5h. The reaction mixture was evaporated and azeotroped with wσpropanol (x3) to yield a dark oil. Purification by chromatography on silica eluting with methanol / dichloromethane (2 : 98) to yielded the sub-titled compound as a dark coloured oil (81 mg).

¹H NMR (400 MHz, CDCl₃) δ 8.82 (2H, dd), 8.06 (IH, d), 7.93 (IH, d), 7.66 (IH, dd), 3.45 (2H, t), 3.05 (2H, t), 2.31 (2H, dt).

Example 30: (_JR)-3-((5)-2-Phenyl-2-piperidin-l-yl-propionyloxy)-l-(3-quinoxalin-6-yl- propyl)-l-azonia-bicyclo[2.2.2]octane bromide

6-(3-Bromo-propyl)-quinoxaline hydrobromide salt (Example 30c) (~80mg ) was treated with 10% sodium hydroxide solution (2 mL) and then extracted with diethyl ether (x3). The organic layer was washed with water, dried and the solvent was evaporated to yield the free base as a dark orange oil. (5)-2-Phenyl-2-piperidin-l-yl -propionic acid (jR)-(l-aza- bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (62 mg) in acetonitrile (0.5 mL) was treated with 6-(3-bromo-propyl)-quinoxaline (50 mg) and the mixture was stirred at room temperature for 18h. The volume of solvent was reduced, diethyl ether was added and the resulting solid was collected by filtration, washed with further diethyl ether and dried to afford the titled compound (65 mg).

m/e 513 [M]⁺

¹H NMR (300 MHz, DMSO) δ 8.94 (2H, dd), 8.09 (IH, d), 7.99 (IH, d), 7.80 (IH, dd), 7.54 (2H, d), 7.32 (2H, t), 7.19 (IH, t), 5.10 - 5.03 (IH, m), 3.88 - 3.76 (IH, m), 3.55 - 3.17 (5H, m), 3.15 - 3.00 (2H, m), 2.86 (2H, t), 2.46 - 2.25 (4H, m), 2.19 (IH, s), 2.11 - 1.71 (6H, m), 1.55 (3H, s), 1.58 - 1.46 (4H, m), 1.46 - 1.37 (2H, m).

Example 31: (l?)-l-[3-(6-Methyl-pyridiπ-3-yl)-propyl]-3-((S)-2-phenyI-2-piperidin-l- yl-propionyloxy)-l-azonia-bicydo[2.2.2]octane bromide

a) 3-(6-Methyl-pyridin-3-yl)-prop-2-yn-l-ol 2008/002072

71

5-Bromo-2-methyl-pyridine (1.5 g) in acetonitrile (25 mL) was treated with propargyl alcohol (0.52 mL) and triethylamine (2.431 mL) followed by copper(I) iodide (0.018 g) and bis(triphenylphosphine)palladium chloride (0.061 g). The mixture was stirred under nitrogen and then heated at 8O⁰C for 48h. The reaction mixture was allowed to cool and filtered. The resulting filtrate was evaporated and then purified by chromatography on silica eluting with ethylacetate / isohexane (3 : 7 to 7 : 3) to yield the sub-titled compound as a yellow solid (0.27 g).

¹H NMR (400 MHz, DMSO) δ 8.49 (IH, d), 7.72 (IH, dd), 7.27 (IH, d), 5.36 (IH, t), 4.31 (2H, d), 2.47 (3H, s).

b) 3-(6-Methyl-pyridin-3-yl)-propan- 1 -ol

3-(6-Methyl-pyridin-3-yl)-prop-2-yn-l-ol (Example 31a) (270 mg) in ethanol (20 mL) was treated with 10% palladium on carbon (0.02 g) in ethanol and hydrogen at 3 bar for 3h with rapid stirring. The reaction mixture was filtered and the catalyst was washed with ethanol. The filtrate was evaporated to yield the sub-titled compound as an oil (270 mg).

¹H NMR (400 MHz, DMSO) 5 8.27 (IH, d), 7.49 (IH, dd), 7.14 (IH, d), 4.48 (IH, t), 3.39 (2H, dd), 2.57 (2H, t), 2.41 (3H, s), 1.72 - 1.64 (2H, m).

c) 5-(3-Bromo-propyl)-2-methyl-pyridine hydrobromide salt

.HBr

3-(6-Methyl-pyridin-3-yl)-propan-l-ol (Example 31b) (0.27 g) in hydrobromic acid (3 mL) was heated at 105⁰C for 3h. The reaction mixture was evaporated and azeotroped with isopropanol (x3) to yield the sub-titled compound as an off-white solid (0.363 g).

¹H NMR (400 MHz, DMSO) δ 8.69 (IH, d), 8.35 (IH, dd), 7.83 (IH, d), 3.55 (2H, t), 2.87 (2H, t), 2.67 (3H, s), 2.22 - 2.14 (2H, m).

Example 31: (_JR)-l-[3-(6-Methyl-pyridin-3-yl)-propyl]-3-((S)-2-phenyl-2-piperidin-l- yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

5-(3-Bromo-propyl)-2-methyl-pyridine hydrobromide salt (Example 31c) (170 mg) was treated with 10% sodium hydroxide solution (2 mL) and then extracted with diethyl ether (x3). The organic layer was washed with brine, dried and the solvent was evaporated to yield the free base as a colourless oil. (S)-2-Phenyl-2-piρeridin-l-yl-propionic acid (.R)-(I- aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (180 mg) in acetonitrile (0.5 mL) was treated with 5-(3-bromopropyl)-2-methylpyridine (146 mg) and the mixture was stirred at room temperature for 18h. The volume of solvent was reduced and the resulting white solid was filtered off, washed with diethyl ether and dried to afford the titled compound (165 mg).

m/e 476 [M]⁺

¹H NMR (400 MHz, DMSO) 5 8.33 (IH, d), 7.58 - 7.52 (3H, m), 7.33 (2H, t), 7.24 - 7.19 (2H, m), 5.08 - 5.03 (IH, m), 3.84 - 3.75 (IH, m), 3.49 - 3.23 (3H, m), 3.23 - 3.12 (2H, m), 3.12 - 2.99 (2H, m), 2.59 - 2.47 (2H, m), 2.44 (3H, s), 2.47 - 2.38 (2H, m), 2.38 - 2.30 (2H, m), 2.18 (IH, s), 1.98 - 1.71 (6H, m), 1.55 (3H, s), 1.57 - 1.48 (4H, m), 1.47 - 1.38 (2H, m).

Example 32: (R)-3-((5)-2-Phenyl-2-piperidin-l-yl-propionyloxy)-l-(3-pyridin-4-yl- propyl)-l-azonia-bicyclo[2.2.2]octane bromide

a) 4-(3-Bronio-propyl)-pyridine hydrobromide

A solution of 3-pyridin-4-yl-propan-l-ol (2.88 mL) in hydrobromic acid (16 mL, 141.43 mmol) was heated at reflux at 135°C for 18h. The cooled solution was concentrated under vacuum and the residue was re-dissolved in wopropanol and re-concentrated (this process was repeated three more times). The residue was dissolved in wopropanol, decolourised by boiling with activated charcoal, filtered, and the clear solution was left to crystallise in a freezer over 48h. The resulting crystals were removed by filtration, washed with wσpropanol / diethyl ether (1:1) followed by diethyl ether and then dried under vacuum at 40⁰C and at room temperature to afford the sub-titled compound as a pale brown solid (3.55 g).

¹H NMR (400 MHz, D₂O): δ 8.64 (2H, d), 7.96 (2H, d), 3.52 (2H, t), 3.12 (2H, t), 2.30 (2H, quint.).

Example 32: (J?)-3-((S)-2-Phenyl-2-piperidin-l-yl-propionyloxy)-l-(3-pyridin-4-yI- propyl)-l-azonia-bicyclo[2.2.2]octane bromide 2008/002072

74

4-(3-Bromo-propyl)-pyridine hydrobromide (Example 32a) (110 mg) was treated with 10% sodium hydroxide solution (2 mL) and then extracted with diethyl ether (x3). The organic layer was washed with brine, dried (MgSO₄) and the solvent was evaporated to yield a colourless residue. The residual free base (68 mg) was added to a solution of (S)-2- phenyl-2-piperidin-l-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (86 mg) in acetonitrile (0.5 mL) and the mixture was stirred at room temperature for 48h. The resulting white solid was filtered off, washed with diethyl ether and dried to afford the titled compound (52 mg).

m/e 462 [M]⁺

¹H NMR (400 MHz, DMSO) δ 8.54 - 8.50 (2H, m), 7.55 (2H, d), 7.34 (2H, t), 7.31 - 7.26 (2H, m), 7.26 - 7.20 (IH, m), 5.10 - 5.03 (IH, m), 3.86 - 3.76 (IH, m), 3.51 - 3.25 (3H, m), 3.24 - 3.14 (2H, m), 3.13 - 3.00 (2H, m), 2.65 - 2.55 (2H, m), 2.47 - 2.39 (2H, m), 2.38 - 2.28 (2H, m), 2.19 (IH, s), 1.99 - 1.71 (6H, m), 1.56 (3H, s), 1.55 - 1.48 (4H, m), 1.47 - 1.38 (2H, m).

Example 33: (R)-l-[3-(2-Methyl-pyridin-4-yl)-propyI]-3-((S)-2-phenyl-2-piperidin-l- yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 4-(3-Bromo-propyl)-2-methyl-pyridine hydrobromide

3-(2-Methyl-pyridin-4-yl)-propan-l-ol (0.5 g) in hydrobromic acid (4 mL, 73.66 mmol) was heated at 105⁰C for 18h. The mixture was evaporated under reduced pressure and azeotroped with zsøpropanol (x6) to yield the sub-titled compound as an oil (0.53 g).

m NMR (400 MHz, DMSO): δ 8.73 (IH, d), 7.87 (IH, s), 7.81 (IH, dd), 3.57 (2H, t), 2.97 (2H, t), 2.70 (3H, s), 2.50 (2H, quint.).

Example 33: (R)-l-[3-(2-Methyl-pyridin-4-yl)-propyI]-3-((S)-2-phenyI-2-piperidin- 1- yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

4-(3-Bromo-propyl)-2-methyl-pyridine hydrobromide (Example 33a) (~200mg wet) was treated with 10% sodium hydroxide solution (2 mL) and then extracted with diethyl ether (x3). The organic layer was washed with brine, dried (MgSO₄) and the solvent was evaporated to yield a colourless oil. The residual free base (114 mg) was added to a solution of (S)-2-phenyl-2-piperidin-l-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ia) (122 mg) in acetonitrile (0.5 mL) and the mixture was stirred at room temperature for 18h. The volume of solvent was reduced under reduced pressure and diethyl ether was added. The solvents were decanted and the product crystallised by addition of acetone and diethyl ether. The resulting white solid was collected by filtration, washed with diethyl ether and dried to afford the titled compound (122 mg).

m/e 476 [M]⁺

¹H NMR (400 MHz, DMSO) δ 8.37 (IH, d), 7.54 (2H, d), 7.33 (2H, t), 7.22 (IH, t), 7.14 (IH, s), 7.07 (IH, d), 5.09 - 5.03 (IH, m), 3.84 - 3.76 (IH, m), 3.50 - 3.00 (7H, m), 2.58 - 2.47 (2H, m), 2.45 (3H, s), 2.47 - 2.38 (2H, m), 2.38 - 2.30 (2H, m), 2.18 (IH, s), 1.96 - 1.82 (4H, m), 1.81 - 1.73 (2H, m), 1.55 (3H, s), 1.57 - 1.47 (4H, m), 1.47 - 1.37 (2H, m).

Pharmacological Analysis

The affinity (pIC₅o) of compounds to the M₃ receptor was determined by competition binding of [³H]N-methyl scopolamine (NMS) to CHO-Kl (Chinese Hamster Ovary) cell membranes expressing the human muscarinic acetylcholine M₃ receptor (M₃-ACh) in a scintillation proximity assay (SPA) format.

SPA beads were precoated with membranes and then incubated at 2mg of beads per well with serial dilutions of the compounds of the invention, [³H]NMS at 0.2nM, half Kd (experimentally determined dissociation constant) and assay buffer (20 mM HEPES pH 7.4 containing 5 mM MgCl₂). The assay was conducted in a final volume of 200 μL, in the presence of 1% (v/v) dimethyl sulphoxide (DMSO). Total binding of [³H]NMS was determined in the absence of competing compound and non-specific binding of [³H]NMS was determined in the presence of 1 μM atropine. The plates were incubated for 16 hours at room temperature and then read on Wallac Microbeta using a normalised H protocol. The PIC₅₀, defined as the negative logarithm of the concentration of compound required for 50% reduction in specific [³H]-NMS binding, was determined. The following table shows the PIC₅₀ figures for representative Examples.

M3 Binding IC₅₀ <2nM "+++"; IC₅₀2-10nM "++"; IC₅₀> 1OnM "+".

Using the procedure given above ρIC₅o values determined for Examples 2, 3, 4, 12, 23 and 33 were 9.8, 9.8, 9.6, 9.6, 9.3 and 9.9 respectively.

The standard deviation of the M₃ receptor assay described above, is between 0.2 and 0.3 log units. The pIC50 values quoted above are means of replicate determinations which were within 2 x SD (95% confidence) of each other.

Measurement of Plasma Protein Binding

The extent of plasma protein binding was determined via equilibrium dialysis of a compound between human plasma and aqueous buffer at 37°C and determination of the concentration of compound in the plasma and buffer by HPLC-MS/MS.

Method

Dialysis cells (molecular weight cut-off 5000) were prepared by rinsing with water followed by soaking in the dialysis buffer for a minimum of 1 hour. The dialysis buffer was isotonic buffered saline pH 7.4. Stock solutions of compound in dimethylsulphoxide were prepared at a concentration of 0.5mM. Frozen pooled Human plasma was obtained from volunteers. The stock DMSO solution of a compound was added to the plasma at a ratio of 10 μl of DMSO to each ml of plasma. This gave a 1% DMSO in plasma solution with each compound at a concentration of 5 μM.

Dialysis cells were then prepared and one half of the cell filled with 750 μl of dialysis buffer and the other half of the cell with 750 μl of plasma solution of compound. Once prepared the cells were sealed and placed in an incubator box at 37⁰C. These cells were then rotated for a minimum of 4 hours to equilibrate.

After equilibration 500 μl of the buffer samples were removed and added to HPLC vials along with 100 μl of plasma (sample in 6-fold diluted plasma), and 100 μl of the plasma samples were removed and added to HPLC vials along with 500 μl of dialysis buffer (sample in 6-fold diluted plasma).

The samples were then analysed using HPLC-MS/MS. A four point calibration curve was obtained by dilutions of the stock solutions with 6-fold diluted plasma at concentrations of 0.013 μM, 0.05 μM, 0.25 μM and 1.25 μM which were injected in this order followed by the buffer sample and then the plasma sample.

Calculation

The concentration of compound in the samples were determined using MassLynx version 4.1 software (produced by Waters/Micromass) that automatically calculated a calibration curve and the concentration of compound in the cells. Plasma protein binding was determined from the calibration curve as the percentage of compound bound in human plasma (% bound) using the following equation;

buffer peak area /

/buffer injection volume % bound = 100- 100 ' ^J

51 plasma peak area.

\ plasma injection volume/ )

For Example 57 the measured human plasma protein binding figure using the procedure described above was 98% bound. Methacholine Induced Bronchoconstriction in vivo

Dunkin-Hartley guinea-pigs (300 - 600g) were supplied by a designated breeding establishment. Animals were dosed with test compound or vehicle either by inhalation in conscious guinea-pigs or by intratracheal instillation (0.5ml/kg) under recoverable gaseous anaesthesia (5% halothane). Animals were allowed to recover from the anaesthesia prior to the measurement of bronchoconstriction. Up to 48 hours post-dosing guinea-pigs were terminally anaesthetized with sodium pentobarbitone (60 mg/kg), the trachea cannulated for artificial ventilation and the jugular vein was cannulated for intravenous administration of methacholine. The guinea-pigs were ventilated using a constant volume respiratory pump (Harvard Rodent Ventilator model 683) at a rate of 60 breath/min and a tidal volume of 5 ml/kg during surgical preparation. Lung function (lung resistance and compliance) was measured in anaesthetised and ventilated guinea-pigs using a pulmonary measurement Flexivent system (SCIREQ, Montreal, Canada) connected to the tracheal cannulae. The animals were ventilated (quasi-sinusoidal ventilation pattern) at 60 breaths/min at a tidal volume of 5 ml/kg. A positive end expiratory pressure of 2-3 CmH₂O was applied. Respiratory resistance was measured using the Flexivent "snapshot" facility (1 second duration, 1 Hz frequency). Lung resistance and compliance was measured before and after intravenous administration of methacholine (3, 10 and 30 ug/kg). The peak increase in resistance following methacholine challenge was calculated and the effect of the test compound on methacholine-induced lung function changes was calculated.

Percentage inhibition of bronchoconstriction was calculated at each dose of methacholine as follows:

[Change in resistance in vehicle treated group - Change in resistance in compound treated group! x 100 [Change in resistance in vehicle treated group]

Inhibition of pilocarpine induced salivation by i.n. administered compounds.

Guinea pigs (450-550g) supplied by Harlan UK or David Hall, Staffs UK and acclimatised to the in-house facilities for a minimum of three days before use. Guinea pigs were randomly assigned into treatment groups and weighed. Each animal was lightly anaesthetised (4% Halothane) and administered compound or vehicle intranasally (0.5ml/kg) at up to 24 hours before challenge with pilocarpine. At the test time point, guinea pigs were terminally anaesthetised with urethane (25% solution in H₂O, 1.5g/kg). Once sufficient anaesthesia had developed (absence of toe pinch reflex) each animal had an absorbent pad placed in the mouth for 5 minutes to dry residual saliva, this pad was removed and replaced with a new pre-weighed pad for 5 minutes to establish a reading of baseline saliva production. At the end of this 5 minute period the pad was removed and weighed. A new pre-weighed pad was inserted into the mouth before each animal received s.c. pilocarpine administered under the skin at the back of the neck (0.6mg/kg @ 2ml/kg). The pad was removed, weighed and replaced with a new pre-weighed pad every 5 minutes up to 15 minutes.

Saliva production was calculated by subtracting the pre-weighed weight of the pad from each 5 minute period post weighed pad and these numbers added together to produce an accumulation of saliva over 15 minutes. Each 5 minute period could be analysed in addition to the whole 15 minute recording period. Baseline production of saliva was assumed to be constant and multiplied by three to produce a reading for baseline saliva production over 15 minutes.

Inhibition of saliva produced by the compound could be calculated by using the following equation: ( l-(Test-baseline)/(Veh-baseline))* 100.

Claims

1. A compound selected from the group consisting of:

(R)-I -[2-(2-Fluoro-phenyl)-ethyl]-3-((5)-2-phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 - azonia-bicyclo[2.2.2]octane X;

(R)- l-[2-(2-Fluoro-phenyl)-ethyl]-3-((.S)-2-piperidin- 1 -yl-2-thioρhen-2-yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(R)- 1 -[2-(2-Fluoro-phenyl)-ethyl]-3-((i?)-2-piperidin- 1 -yl-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(R)-I -(6-Methyl-benzooxazol-2-ylmethyl)-3-((5)-2-phenyl-2-piperidin-l -yl- propionyloxy)- 1 -azonia-bicyclo[2.2.2]octane X;

(R)- 1 -(3-Benzooxazol-2-yl-propyl)-3-((>S)-2-phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 - azonia-bicyclo[2.2.2]octane X; (R)-I -(5-Chloro-benzooxazol-2-ylmethyl)-3-((S)-2-phenyl-2-piperidin- 1 -yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(R)-l-(6-Chloro-benzooxazol-2-ylmethyl)-3-((5)-2-phenyl-2-piperidin-l-yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(R)- 1 -Benzooxazol-2-ylmethyl-3-((₁S)-2-phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 - azonia-bicyclo[2.2.2]octane X;

(R)-l-(5-Methyl-benzooxazol-2-ylmethyl)-3-((5)-2-phenyl-2-piperidin-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(R)-3-((S)-2-Phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 -( 1 -phenyl- 1 H-pyrazol-4- ylmethyl)-l-azonia-bicyclo[2.2.2]octane X; (R)-3-((S)-2-Phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 -( 1 -phenyl- lH-[ 1 ,2,3]triazol-4- ylmethyl)-l-azonia-bicyclo[2.2.2]octane X;

(R)- 1 -Isopropyl-3-((5')-2-phenyl-2-piρeridin- 1 -yl-propionyloxy)- 1 -azonia- bicyclo[2.2.2]octane X;

(R)-l-(5-Phenyl-[l,2,4]oxadiazol-3-ylmethyl)-3-((5)-2-piperidin-l-yl-2-thioρhen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(R)-l-(5-Phenyl-[l,2,4]oxadiazol-3-ylmethyl)-3-((R)-2-piperidin-l-yl-2-thioρhen-2-yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X; (i?)-l-(3-Phenyl-isoxazol-5-ylmethyl)-3-((5')-2-piperidin-l-yl-2-thiophen-2-yl- ρropionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(R)- l-(3-Phenyl-isoxazol-5-ylmethyl)-3-((/?)-2-piperidin- 1 -yl-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X; (i?)-l-[3-(4-Methoxy-phenyl)-[l,2,4]oxadiazol-5-ylmethyl]-3-((S)-2-piρeridin-l-yl-2- thiophen-2-yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(R)- 1 -[3-(4-Methoxy-phenyl)-[ 1 ,2,4]oxadiazol-5-ylmethyl]-3-((i?)-2-piperidin- 1 -yl-2- thiophen-2-yl-propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(/?)-l-[3-(5-Methyl-pyridin-3-yl)-proρyl]-3-((S)-2-piperidin-l-yl-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(i?)-l-[3-(5-Methyl-pyridin-3-yl)-propyl]-3-((i?)-2-ρiperidin-l-yl-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(/?)-l-(5-Phenyl-isoxazol-3-ylmethyl)-3-((.S')-2-piperidin-l-yl-2-thiophen-2-yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X; (/?)-l-(5-Phenyl-isoxazol-3-ylmethyl)-3-((i?)-2-piperidin-l-yl-2-thiophen-2-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(R)- 1 -{ 2-[4-(2-Hydroxy-ethyl)-phenyl] -ethyl } -3-((5)-2-phenyl-2-piperidin- 1-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(i?)-l-[2-(lH-Indol-3-yl)-ethyl]-3-((5')-2-phenyl-2-ρiperidin-l-yl-propionyloxy)-l- azonia-bicyclo[2.2.2]octane X;

(R)- 1 -(5-Phenyl-oxazol-2-ylmethyl)-3-((5)-2-phenyl-2-piperidin- 1 -yl-propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(i?)-l-(5-Phenyl-isoxazol-3-ylmethyl)-3-((5)-2-phenyl-2-piperidin-l-yl-propionyloxy)- l-azonia-bicyclo[2.2.2]octane X; (R)- 1 -(3-Phenyl-isoxazol-5-ylmethyl)-3-((S)-2-phenyl-2-piperidin- 1 -yl-propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(^-l-CS-Phenyl-tl^^oxadiazol-S-ylmethy^-S-Crø^-phenyl^-piperidin-l-yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(R)- 1 -(5-Phenyl-[ 1 ,2,4]oxadiazol-3-ylmethyl)-3-((5)-2-phenyl-2-piperidin- 1-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(R)-l-(2-Phenyl-oxazol-4-ylmethyl)-3-((_lS')-2-phenyl-2-piperidin-l-yl-propionyloxy)- l-azonia-bicyclo[2.2.2]octane X; (^-l-tS-CS-Fluoro-phenyO^l^^Joxadiazol-S-ylmethy^-S-Crø-l-phenyl-l-piperidin- l-yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(R)- 1 -(2-Phenyl-oxazol-5-ylmethyl)-3 -((5>2-phenyl-2-piperidin- 1 -yl-propionyloxy)- l-azonia-bicyclo[2.2.2]octane X; (i?)-3-((5')-2-Phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 -(3-pyridin-4-yl-

[l,2,4]oxadiazol-5-ylmethyl)-l-azonia-bicyclo[2.2.2]octane X;

(R)- 1 - { 2-[2-(4-Chloro-phenyl)-thiazol-4-yl]-ethyl } -3-((5')-2-phenyl-2-piperidin- 1 -yl- propionyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(i?)-l-(3-Benzo[b]thiophen-2-yl-propyl)-3-((5')-2-phenyl-2-piperidin-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(i?)-3-((5)-2-Phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 -(3-quinoxalin-6-yl-propyl)- 1 - azonia-bicyclo[2.2.2]octane X;

(R)- l-[3-(6-Methyl-pyridin-3-yl)-propyl]-3-((_ιS')-2-ρhenyl-2-piperidin- 1 -yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X; (i?)-3-((S)-2-Phenyl-2~piperidin- 1 -yl-propionyloxy)- 1 -(3-pyridin-4-yl-propyl)- 1 - azonia-bicyclo[2.2.2]octane X; and

(i?)-l-[3-(2-Methyl-pyridin-4-yl)-propyl]-3-((.S)-2-phenyl-2-piperidin-l-yl- propionyloxy)-l-azonia-bicyclo[2.2.2]octane X;

wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid.

2. A pharmaceutical composition comprising a compound according to claim 1 in association with a pharmaceutically acceptable adjuvant, diluent or carrier.

3. A process for the preparation of a pharmaceutical composition as claimed in claim 2, which comprises mixing a compound according to claim 1 with a pharmaceutically acceptable adjuvant, diluent or carrier.

4. A compound according to claim 1 for use in therapy.

5. Use of a compound according to claim 1 in the manufacture of a medicament for use in the treatment of chronic obstructive pulmonary disease.

6. A method of treating chronic obstructive pulmonary disease in a warm-blooded animal, such as man, which comprises administering to a mammal in need of such treatment an effective amount of a compound according to claim 1.

7. A pharmaceutical product comprising, in combination, a first active ingredient which is a compound according to claim 1 and at least one further active ingredient selected from:-

• a phosphodiesterase inhibitor,

• a β2. adrenoceptor agonist,

• a modulator of chemokine receptor function,

• an inhibitor of kinase function, • a protease inhibitor,

• a steroidal glucocorticoid receptor agonist, and a

• a non-steroidal glucocorticoid receptor agonist.