QUINUCLIDINE DERIVATIVES AS MUSCARINIC M3 RECEPTOR ANTAGONISTS

NEW COMPOUNDS 273

The present invention relates to cycloalkyl-substituted alkyl esters of polycyclic amine alcohols, a process for their preparation, pharmaceutical compositions containing then process for preparing pharmaceutical compositions, their use in therapy and intermedi of use in their preparation.

Background to the Invention

Muscarinic receptors are a G-protein coupled receptor (GPCR) family having five farr members Mi, M ₂ , M ₃ , M ₄ and M ₅ . Of the five muscarinic subtypes, three (Mj, M ₂ and 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 chπ obstructive pulmonary disease (COPD), and for this reason muscarinic receptor antagc have been developed for use in treating airway diseases. Muscarinic receptor antagoni often called anticholinergics in clinical practice, have gained widespread acceptance a first-line therapy for individuals with COPD, and their use has been extensivley reviev 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 maj( of muscarinic antagonists have a relatively short duration of action requiring that they 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 d 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 1 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 dail> dosing.

WO 98/04517 describes arylcyclopropane, arylcyclobutane, arylcyclopentane and arylcyclohexane carboxylic esters having antimuscarinic activity on the urinary bladde smooth muscle.

Our co-pending application PCT/GB2007/004350 relates to compounds of formula (I)

wherein

R ¹ and R ² together with the carbon atom to which they are both directly attached form membered aliphatic carbocyclic ring which may be optionally substituted by one or m substituents independently selected from halogen, hydroxyl, Ci _-6 alkoxy, NH ₂ , NH(Ci alkyl), N(Ci _-6 alkyl) ₂ and Ci _-6 alkyl which Ci _-6 alkyl may be optionally substituted by or more substituents independently selected from halogen and hydroxyl;

R ³ 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 haloge cyano, nitro, SH, S(O) _0-2 R ⁹ , NR ¹⁰ R ¹¹ , S(O) ₂ NR ¹² R ¹³ , C(O)NR ¹⁴ R ¹⁵ , C(O) ₂ R ¹⁶ , NR ¹⁷ S(O) ₂ R ¹⁸ , NR ¹⁹ C(O)R ²⁰ , NR ²¹ C(O) ₂ R ²² , NR ²³ C(O)NR ²⁴ R ²⁵ , OR ²⁶ and C _1-6 alkyl which Ci _-6 alkyl may be optionally substituted by one or more substituents independei selected from halogen, hydroxyl, Ci _-6 alkoxy, NH ₂ , NH(Ci _-6 alkyl) and N(Ci _-6 alkyl) ₂ ; R ⁴ represents a group of formula (II) or (Ilia) or (HIb);

wherein

Y is -CH ₂ -, -CH ₂ CH ₂ - or -CH ₂ CH ₂ CH ₂ - and the substitution on the ring in group (II) r be in the 3 or 4 positions; a is 1 or 2; b is 1 or 2; Z is -CH ₂ -; R ⁵ represents a group of formula (IV)

wherein w is 0 or 1 ;

R ⁶ represents Ci _-4 alkylene optionally substituted by one or more substituents independently selected from halogen, hydroxyl, Ci _-6 alkoxy, NH?, NH(Q _-6 alkyl) and N(C _1-6 alkyl) ₂ ; when w is 0, y is 0; when w is 1 , y is 0 or 1 ;

Q represents O, S(O) _0-2 , NR ⁸ , -CONR ⁸ -, -SO ₂ NR ⁸ -, -NR ⁸ CO-, -NR ⁸ SO ₂ -, -OC(O)-, -

C(O)O-, -HC=CH- or ethynylene;

R ⁷ represents a cyclic group Cyc ¹ or a Ci _-4 alkyl group which Ci _-4 alkyl group may be optionally substituted by one or more substituents independently selected from haloge hydroxyl, Q _-4 alkoxy, NH ₂ , NH(Ci _-4 alkyl), N(Ci _-4 alkyl) ₂ , a cyclic group Cyc ² and -

OCyc ² ; and R ⁷ may additionally represent hydrogen when Q represents O, NR ⁸ , -COPv

-SO ₂ NR ⁸ -, -C(O)O-, -HC=CH- or ethynylene;

Cyc ¹ and Cyc each independently represent aryl, heteroaryl, a 3 to 8 membered aliph; carbocyclic.ring or a 4 to 8 membered aliphatic heterocyclic ring, each of which may [ optionally substituted by one or more substituents independently selected from haloge

alkyl which phenyl or Ci_ ₆ alkyl may be optionally substituted by one or more substiti independently selected from halogen, hydroxyl, C _f _ ₆ alkoxy, NH ₂ , NH(Ci_ ₆ alkyl) and N(C _1-6 alkyl) ₂ ;

R ⁸ represents hydrogen or Ci _-6 alkyl; R ⁹ and R ¹⁸ each independently represent Cj _-6 alkyl, which Ci _-6 alkyl may be optionall; substituted by one or more substituents independently selected from halogen, hydrox> ₆ alkoxy, NH ₂ , NH(C _1-6 alkyl) and N(C _1-6 alkyl) ₂ ; and

R ¹⁰ , R ^{1 1} , R ¹² , R ¹³ , R ¹⁴ , R ¹⁵ , R ¹⁶ , R ¹⁷ , R ¹⁹ , R ²⁰ , R ²¹ , R ²² , R ²³ , R ²⁴ , R ²⁵ and R ²⁶ each independently represent hydrogen or Ci _-6 alkyl, which Ci- ₆ alkyl may be optionally substituted by one or more substituents independently selected from halogen, hydroxy ₆ alkoxy, NH ₂ , NH(C _1-6 alkyl) and N(C _1-6 alkyl) ₂ ; or any of R ¹⁰ and R ^{1 1} , R ¹² and R ¹³ , R R ¹⁵ or R ²⁴ and R ²⁵ , together with the nitrogen atom to which they are both attached, m form a 4 to 8 membered aliphatic heterocyclic ring, which heterocyclic ring may be optionally substituted by one or more substituents independently selected from haloge hydroxyl and Ci _-6 alkyl, which C _1-6 alkyl may be optionally substituted by one or mon 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 specific disclosed in, our co-pending application PCT/GB2007/004350 referred to above.

Thus, the present invention provides a compound which has a quaternary ammonium species selected from the group consisting of:

(/?)-l-[(6-Methyl-pyridin-3-ylcarbamoyl)-methyl]-3-(l-phe nyl-cycloheptanecarbonyk l-azonia-bicyclo[2.2.2]octane X;

(R)- l-[(6-Methyl-pyrazin-2-ylcarbamoyl)-methyl]-3-( 1 -phenyl-cycloheptanecarbonyh l-azonia-bicyclo[2.2.2]octane X;

(/?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-[(6-trifluoro methyl-pyridazin-3-

(R)- 1 -(Benzo[<i]isoxazol-3-ylcarbamoylmethyl)-3-( 1 -phenyl-cycloheptanecarbonylox azonia-bicyclo[2.2.2]octane X;

(R)- 1 -(Pyridazin-3-ylcarbamoylmethyl)-3-( 1 -thiophen-2-yl-cycloheptanecarbonyloxy azonia-bicyclo[2.2.2]octane X; (R)-l-[(5-Methyl-isoxazol-3-ylcarbamoyl)-methyl]-3-(l-thioph en-2-yl- cycloheptanecarbonyloxy)- 1 -azonia-bicyclo[2.2.2]octane X;

(R)- 1 -[(3-Methyl-isoxazol-5-ylcarbamoyl)-methyl]-3-( 1 -thiophen-2-yl- cycloheptanecarbonyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(R)-l-[(3-Fluoro-phenylcarbamoyl)-methyl]-3-(l-thiophen-2 -yl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane X;

(R)-l-[(5-Methyl-pyrazin-2-ylcarbamoyl)-methyl]-3-(l-thio phen-2-yl- cycloheptanecarbonyloxy)- 1 -azonia-bicyclo[2.2.2]octane X;

(R)-l-(Benzo[<i]isoxazol-3-ylcarbamoylmet:hyl)-3-(l-th iophen-2-yl- cycloheptanecarbonyloxy)- l-azonia-bicyclo[2.2.2]octane X; (R)-I -(Pyrazin-2-ylcarbamoylmethyl)-3-(l-thiophen-2-yl-cyclohepta necarbonyloxy)-] azonia-bicyclo[2.2.2]octane X;

(R)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(py razin-2-ylcarbamoylmeth l-azonia-bicyclo[2.2.2]octane X;

(R)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(is oxazol-3-ylcarbamoylmetl l-azonia-bicyclo[2.2.2]octane X;

(R)-3-( 1 -Phenyl -cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)- 1 -azoni; bicyclo[2.2.2]octane X;

(R)-3-( 1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-4-ylcarbamoylmethyl)- 1 -azonii bicyclo[2.2.2]octane X; (R)-l-[(5-Fluoro-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonylo l-azonia-bicyclo[2.2.2]octane X;

(R)-l-[(5-Methyl-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phen yl-cycloheptanecarbonylc l-azonia-bicyclo[2.2.2]octane X;

(R)-3-( 1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-3-ylcarbamoylmethyl)- 1 -azonii bicyclo[2.2.2]octane X;

(R)-l-[(2-Methyl-pyridin-4-ylcarbamoyl)-methyl]-3-(l-phen yl-cycloheptanecarbonylc

(R)- 1 -Phenylcarbamoylmethyl-3-( 1 -phenyl-cycloheptanecarbonyloxy)- 1 -azonia- bicyclo[2.2.2]octane X;

(R)-3-( 1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrimidin-4-ylcarbamoylmethyl)- 1 -azo bicyclo[2.2.2]octane X; (R)-I -[(2-Fluoro-phenylcarbamoyl)-methyl]-3-(l -phenyl-cycloheptanecarbonyloxy)- 1 azonia-bicyclo[2.2.2]octane X;

(R)-l-[(2,3-Difluoro-phenylcarbamoyl)-methyl]-3-(l-phenyl -cycloheptanecarbonylox; azonia-bicyclo[2.2.2]octane X;

(R)-l-[2-(2,3-Dihydro-benzofuran-5-yl)-ethyl]-3-(l-phenyl -cycloheptanecarbonyloxy) azonia-bicyclo[2.2.2]octane X;

(R)- l-[2-(4-Fluoro-phenoxy)-ethyl]-3-(l -phenyl-cycloheptanecarbonyloxy)- 1-azonia- bicyclo[2.2.2]octane X;

(R)-3-( 1 -phenyl-cycloheptanecarbonyloxy)- 1 -(pyridazin-4-ylcarbamoylmethyl)- 1 -azoi bicyclo[2.2.2]octane X; (R)-l-[(5-Fluoro-pyridin-3-ylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonylo: l-azonia-bicyclo[2.2.2]octane X;

(R)-3-(l -phenyl-cycloheptanecarbonyloxy)- 1 -[2-(pyridin-3-yloxy)-ethyl]- 1-azonia- bicyclo[2.2.2]octane X;

(R)- 1 -[(6-Methyl-pyridin-2-ylcarbamoyl)-methyl]-3-( 1 -phenyl-cycloheptanecarbonylo l-azonia-bicyclo[2.2.2]octane X;

(R)-3-( 1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(o-tolylcarbamoyl-methyl)- 1 -azonia- bicyclo[2.2.2]octane X;

(R)-3-( 1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(2-pyrazin-2-yl-ethyl)- 1 -azonia- bicyclo[2.2.2]octane X; (S)- 1 -(3-Phenoxy-propyl)-3-( 1 -phenyl-cycloheptanecarbonyloxy)- 1 -azonia- bicyclo[2.2.2]octane X;

(R)-l-{ [2-(3-Fluoro-phenoxy)-ethylcarbamoyl]-methyl}-3-(l-phenyl- cycloheptanecarbonyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(R)-l-[(3,5-Difluoro-phenylcarbamoyl)-methyl]-3-(l-phenyl -cycloheptanecarbonylox] azonia-bicyclo[2.2.2]octane X;

(R)-l-[2-(4-methoxy-benzyloxy)-ethyl]-3-(l-phenyl-cyclohe ptanecarbonyloxy)-l-azor

(R)- 1 -(2-Phenethyloxy-ethyl)-3-( 1 -phenyl-cycloheptanecarbonyloxy)- 1 -azonia- bicyclo[2.2.2]octane X;

(R)-l-[(2,6-Difluoro-phenylcarbamoyl)-methyl]-3-(l-phenyl -cycloheptanecarbonylox azonia-bicyclo[2.2.2]octane X; (/?)-l-[(Methyl-phenyl-carbamoyl)-methyl]-3-(l -phenyl-cycloheptanecarbonyloxy)- 1 azonia-bicyclo[2.2.2]octane X;

(R)- 1 -[3-(4-Cyano-phenoxy)-propyl]-3-( 1 -phenyl-cycloheptanecarbonyloxy)- 1 -azonύ bicyclo[2.2.2]octane X;

(R)-l-[(2,5-Difluoro-phenylcarbamoyl)-methyl]-3-(l-phenyl -cycloheptanecarbonylox azonia-bicyclo[2.2.2]octane X;

(R)- 1 -[2-(4-Cyano-benzyloxy)-ethyl]-3-( 1 -phenyl-cycloheptanecarbonyloxy)- 1 -azoni; bicyclo[2.2.2]octane X;

(R)-3-(l -Phenyl-cycloheptanecarbonyloxy)- l-[(6-trifluoromethyl-pyridin-2-ylcarbam( methyl]- l-azonia-bicyclo[2.2.2]octane X; (/?)-l-[(4-Methyl-pyridin-2-ylcarbamoyl)-methyl]-3-(l -phenyl -cycloheptanecarbonyk l-azonia-bicyclo[2.2.2]octane X;

(/?)-l-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phe nyl-cycloheptanecarbonylc l-azonia-bicyclo[2.2.2]octane X;

(R)-3-( 1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(p-tolylcarbamoyl-methyl)- 1 -azonia- bicyclo[2.2.2]octane X;

(R)-3-( 1 -Phenyl-cycloheptanecarbonyloxy)- l-(/n-tolylcarbamoyl-methyl)- l-azonia- bicyclo[2.2.2]octane X;

(/?)-l-(Oxazol-2-ylcarbamoylmethyl)-3-(l -phenyl-cycloheptanecarbonyloxy)- 1-azonii bicyclo[2.2.2]octane X; (R)- l-[(6-Methyl-pyridazin-3-ylcarbamoyl)-methyl]-3-(l -phenyl- cycloheptanecarbonyloxy)- 1 -azonia-bicyclo[2.2.2]octane X;

(/?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- l-(pyrimidin-2-ylcarbamoylmethyl)-l-azc bicyclo[2.2.2]octane X;

(i?)-l-[(5-Cyano-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phen yl-cycloheptanecarbonylo l-azonia-bicyclo[2.2.2]octane X;

(R)S-(I -Phenyl -eye loheptanecarbonyloxy)-l-(pyrimidin-5-ylcarbamoylmethyl)-l-az c

(i?)-l-[(3-Fluoro-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phe nyl-cycloheptanecarbonylo l-azonia-bicyclo[2.2.2]octane X;

(/?)-l-[(3-Fluoro-pyridin-4-ylcarbamoyl)-methyl]-3-(l-phe nyl-cycloheptanecarbonylo l-azonia-bicyclo[2.2.2]octane X; (/?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-{2-[(pyrazine-2- carbonyl)-amino]-ethyl azonia-bicyclo[2.2.2]octane X;

(R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-([l,2,4]thiadi azol-5-ylcarbamoylmethyl azonia-bicyclo[2.2.2]octane X;

(R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-{3-[(pyridine- 2-carbonyl)-amino]-prop) azonia-bicyclo[2.2.2]octane X;

(/?)-l-[(2-Methyl-pyrimidin-4-ylcarbamoyl)-methyl]-3-(l-p henyl- cycloheptanecarbonyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(R)- 1 -[(6-Methyl-pyrimidin-4-ylcarbamoyl)-methyl]-3-( 1 -phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane X; (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-{2-[(pyridine-2-c arbonyl)-amino]-ethyl azonia-bicyclo[2.2.2]octane X; and

(/?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(3-pyridin-4- yl-propyl)-l-azonia- bicyclo[2.2.2]octane X; wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent aci

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 at( The anion X may be any pharmaceutically acceptable anion of a mono or polyvalent (< bivalent) acid. In an embodiment of the invention X may be an anion of a mineral acid example chloride, bromide, iodide, sulfate, nitrate or phosphate; or an anion of a suital organic acid, for example acetate, maleate, fumarate, citrate, oxalate, succinate, tartrat< methanesulphonate, p-toluenesulphonate, benzenesulphonate, napadisylate (naphthalei 1,5-disulfonate) (e.g. a heminapadisylate), 2,5-dichlorobenzenesulphonate, 1- hydroxynaphthalene-2-sulphonate or xinafoate (l-hydroxy-2-naphthoate).

According to the invention, there is also provided a compound selected from the group.

H

nd

H wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent aci

It will be understood that certain compounds of the present invention may exist in sob for example hydrated, as well as unsolvated forms. It is to be understood that the prest invention encompasses all such solvated forms. Certain compounds of the present invention may exist as tautomers. Tautomers and mixtures thereof also form an aspect the present invention.

The compounds of the present invention display beneficial pharmaceutical properties, example, the compounds of the invention display activity as antagonists of muscarinic receptors, particularly muscarinic M ₃ receptors. Moreover, the compounds also disph desirable plasma protein binding properties. Plasma protein binding may be an advantageous property for compounds administered via inhalation as it can lessen the impact of any systemic effect the compound may have.

The compounds of the invention have activity as pharmaceuticals, in particular as

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 aspi: and NSAID-induced) and dust-induced asthma, both intermittent and persistent and ol severities, and other causes of airway hyper-responsiveness; chronic obstructive pulrm disease (COPD); bronchitis, including infectious and eosinophilic bronchitis; emphysc 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, i iatrogenic cough; acute and chronic rhinitis including rhinitis medicamentosa, and vasomotor rhinitis; perennial and seasonal allergic rhinitis including rhinitis nervosa (1 fever); nasal polyposis; acute viral infection including the common cold, and infection to respiratory syncytial virus, influenza, coronavirus (including SARS) and adenoviru:

2. bone and joints: arthritides associated with or including osteoarthritis/osteoarthro; both primary and secondary to, for example, congenital hip dysplasia; cervical and lur spondylitis, and low back and neck pain; rheumatoid arthritis and Still's disease; seronegative spondyloarthropathies including ankylosing spondylitis, psoriatic arthriti reactive arthritis and undifferentiated spondarthropathy; septic arthritis and other infec related arthopathies and bone disorders such as tuberculosis, including Potts' disease Poncet's syndrome; acute and chronic crystal-induced synovitis including urate gout, calcium pyrophosphate deposition disease, and calcium apatite related tendon, bursal < 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; inflammator; myopathies including dermatomyositits and polymyositis; polymalgia rheumatica; juv arthritis including idiopathic inflammatory arthritides of whatever joint distribution an

including giant cell arteritis, Takayasu's arteritis, Churg-Strauss syndrome, polyarterit 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 example sports injury] or disease: arthitides (for example rheumatoid arthritis, osteoarthritis, gout or crystal arthropathy), other joint disease (such as intervertebral di degeneration or temporomandibular joint degeneration), bone remodelling disease (sue 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 dermatos< and delayed-type hypersensitivity reactions; phyto- and photodermatitis; seborrhoeic dermatitis, dermatitis herpetiformis, lichen planus, lichen sclerosus et atrophica, pyode 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-Chris 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 conjunctiv 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;

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

7. abdominal: hepatitis, including autoimmune, alcoholic and viral; fibrosis and cirri

8. genitourinary: nephritis including interstitial and glomerulonephritis; nephrotic syndrome; cystitis including acute and chronic (interstitial) cystitis and Hunner's ulcei 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; chronic graft versus host disease;

10. CNS: Alzheimer's disease and other dementing disorders including CJD and nvCJ amyloidosis; multiple sclerosis and other demyelinating syndromes; cerebral atherosclerosis and vasculitis; temporal arteritis; myasthenia gravis; acute and chronic (acute, intermittent or persistent, whether of central or peripheral origin) including vis< pain, headache, migraine, trigeminal neuralgia, atypical facial pain, joint and bone paii pain arising from cancer and tumor invasion, neuropathic pain syndromes including diabetic, post-herpetic, and HIV-associated neuropathies; neurosarcoidosis; central an< peripheral nervous system complications of malignant, infectious or autoimmune processes;

11. other auto-immune and allergic disorders including Hashimoto's thyroiditis, Grav 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;

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 aortiti including infective (for example syphilitic); vasculitides; disorders of the proximal anc peripheral veins including phlebitis and thrombosis, including deep vein thrombosis ai 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 affectii 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 co 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 inventi as hereinbefore defined, for use in therapy.

In another aspect, the invention provides the use of a compound of the present inventi< 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 "prophylax unless there are specific indications to the contrary. The terms "therapeutic" and "therapeutically" should be construed accordingly.

A further aspect of the invention provides a method of treating a disease state in a mar suffering from, or at risk of, said disease, which comprises administering to a mamma need of such treatment a therapeutically effective amount of a compound of the preser invention, as hereinbefore defined.

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

The present invention also provides a compound of the present invention, as hereinbef defined, for treating asthma.

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

The present invention also provides the use of a compound of the present invention, a: hereinbefore defined, in the treatment of asthma.

The present invention also provides the use of a compound of the present invention, a: 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, a: 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 pulmo 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 an of a compound of the present invention, as hereinbefore defined.

The present invention further provides a method of treating asthma in a warm-bloodec 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 warmblooded animal, such as man, said ingredient is normally formulated in accordance wi standard pharmaceutical practice as a pharmaceutical composition.

Therefore in another aspect the present invention provides a pharmaceutical compositi that comprises a compound of the invention as hereinbefore defined and a

invention provides a process for the preparation of said composition, which comprise* mixing active ingredient with a pharmaceutically acceptable adjuvant, diluent or carri 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 809i 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 (sucl to the lung and/or airways or to the skin), oral, rectal or parenteral administration. Foi these purposes the compounds of this invention may be formulated by means known i 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 aqu 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 0.1 mg 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 lOOmg of the compound, for example in the range of 0. lmgkg ^"1 to 20mgkg ^" ' of this invention composition being administered 1 to 4 times per day. The intravenous, subcutaneous 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. Alternal each patient will receive a daily oral dose, which is approximately equivalent to the dc parenteral dose, the composition being administered 1 to 4 times per day

Another suitable pharmaceutical composition of this invention is one suitable for inha

compounds of the invention when treating respiratory diseases such as chronic obstruc pulmonary disease (COPD) or asthma. When administered by inhalation the compoun the present invention may be used effectively at doses in the μg range, for example 0.1 500 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, 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 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 inhal administration.

When administered by inhalation, metered dose inhaler devices may be used to admin the active ingredient, dispersed in a suitable propellant and with or without additional excipients such as ethanol, surfactants, lubricants or stabilising agents. Suitable propel include hydrocarbon, chlorofluorocarbon and hydrofluoroalkane (e.g. heptafluoroalkai propellants, or mixtures of any such propellants. Preferred propellants are P 134a and F 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, solut may also be employed, with or without a suitable pH and/or tonicity adjustment, eithei 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 fin divided powder or as an ordered mixture. The dry powder inhaler may be single dose ( 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, is administered concurrently or sequentially or as a combined preparation v 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 rheumatoid arthritis, osteoarthritis, asthma, allergic rhinitis, chronic obstructive pulrnc 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 systemicall) (such as piroxicam, diclofenac, propionic acids such as naproxen, flurbiprofen, fenopi ketoprofen and ibuprofen, fenamates such as mefenamic acid, indomethacin, sulindac azapropazone, pyrazolones such as phenylbutazone, salicylates such as aspirin); selec 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, intravenou intra-articular routes); methotrexate; leflunomide; hydroxychloroquine; d-penicillamii auranofin or other parenteral or oral gold preparations; analgesics; diacerein; intra-arti 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, (inch agents which act on cytokine signalling pathways such as modulators of the SOCS sy: including alpha-, beta-, and gamma-interferons; insulin-like growth factor type I (IGF 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 monocloi antibodies (for example infliximab; adalimumab, and CDP-870) and TNF receptor antagonists including immunoglobulin molecules (such as etanercept) and low-moleci weight agents such as pentoxyfylline.

In addition the invention relates to a combination of a compound of the invention with monoclonal antibody targeting B-Lymphocytes (such as CD20 (rituximab), MRA-alL 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 CX

(for the C-X-C family) and CX ₃ CRl for the C-X ₃ -C family.

The present invention further relates to the combination of a compound of the inventic with an inhibitor of matrix metalloprotease (MMPs), i.e., the stromelysins, the collagenases, and the gelatinases, as well as aggrecanase; especially collagenase- 1 (M 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 lipoxygenase activating protein (FLAP) antagonist such as; zileuton; ABT-761; fenleu tepoxalin; Abbott-79175; Abbott-85761; a N-(5-substituted)-thiophene-2- alkylsulfonamide; 2,6-di-tert-butylphenolhydrazones; a methoxytetrahydropyrans sue! Zeneca ZD-2138; the compound SB-210661; a pyridinyl-substituted 2-cyanonaphthal< compound such as L-739,010; a 2-cyanoquinoline compound such as L-746,530; or ar 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 inventic and a receptor antagonist for leukotrienes (LT) B4, LTC4, LTD4, and LTE4 selected i the group consisting of the phenothiazin-3-ls such as L-651,392; amidino compounds as CGS-25019c; benzoxalamines such as ontazolast; benzenecarboximidamides such < BIIL 284/260; and compounds such as zafirlukast, ablukast, montelukast, pranlukast,

The present invention still further relates to the combination of a compound of the invention and a phosphodiesterase (PDE) inhibitor such as a methylxanthanine includi theophylline and aminophylline; a selective PDE isoenzyme inhibitor including a PDE 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 inventic 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 hista type 2 receptor antagonist.

The present invention further relates to the combination of a compound of the inventic 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- I/alpha- 2 adrenoceptor agonist vasoconstrictor sympathomime agent, such as propylhexedrine, phenylephrine, phenylpropanolamine, ephedrine, pseudoephedrine, naphazoline hydrochloride, oxymetazoline hydrochloride, tetrahydrozoline hydrochloride, xylometazoline hydrochloride, tramazoline hydrochlo 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 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 inventit 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 inventic 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 inventic and another systemic or topically-applied anti-inflammatory agent, such as thalidomid 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 sulfasalazin 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 inventic together with an antibacterial agent such as a penicillin derivative, a tetracycline, a macrolide, a beta-lactam, a fluoroquinolone, metronidazole, an inhaled aminoglycosid antiviral agent including acyclovir, famciclovir, valaciclovir, ganciclovir, cidofovir, amantadine, rimantadine, ribavirin, zanamavir and oseltamavir; a protease inhibitor su indinavir, nelfinavir, ritonavir, and saquinavir; a nucleoside reverse transcriptase inhib such as didanosine, lamivudine, stavudine, zalcitabine or zidovudine; or a non-nucleos

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 angioten 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 platelet aggregation inhibitor.

The present invention further relates to the combination of a compound of the inventk and a CNS agent such as an antidepressant (such as sertraline), an anti-Parkinsonian d (such as deprenyl, L-dopa, ropinirole, pramipexole, a MAOB inhibitor such as selegin 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 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 o peripherally-acting analgesic (for example an opioid or derivative thereof), carbamaze phenytoin, sodium valproate, amitryptiline or other anti-depressant agent-s, paracetam or a non-steroidal anti-inflammatory agent.

The present invention further relates to the combination of a compound of the inventk together with a parenterally or topically-applied (including inhaled) local anaesthetic i 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 as alendronate.

The present invention still further relates to the combination of a compound of the

antagonist; (iii) interleukin converting enzyme (ICE) inhibitor; (iv) IMPDH inhibitor; 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 Imatinib mesylate), a serine / threonine kinase (such as an inhibi a MAP kinase such as p38, JNK, protein kinase A, B or C, or IKK), or a kinase invoh cell cycle regulation (such as a cylin dependent kinase); (viii) glucose-6 phosphate dehydrogenase inhibitor; (ix) kinin-Bl. - or B2. -receptor antagonist; (x) anti-gout age for example colchicine; (xi) xanthine oxidase inhibitor, for example allopurinol; (xii) uricosuric agent, for example probenecid, sulfinpyrazone or benzbromarone; (xiii) grc hormone secretagogue; (xiv) transforming growth factor (TGFβ); (xv) platelet-derivec growth factor (PDGF); (xvi) fibroblast growth factor for example basic fibroblast gro\ factor (bFGF); (xvii) granulocyte macrophage colony stimulating factor (GM-CSF); (: 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; ( TNF- alpha converting enzyme inhibitor (TACE); (xxii) induced nitric oxide synthase (iNOS) inhibitor; (xxiii) chemoattractant receptor-homologous molecule expressed on cells, (such as a CRTH2 antagonist); (xxiv) inhibitor of P38; (xxv) agent modulating t function of Toll-like receptors (TLR), (xxvi) agent modulating the activity of purinerg 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 therap( agent for the treatment of cancer, for example suitable agents include:

(i) an antiproliferative/antineoplastic drug or a combination thereof, as used in medica 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 1 5-fluorouracil or tegafur, raltitrexed, methotrexate, cytosine arabinoside, hydroxyurea gemcitabine or paclitaxel); an antitumour antibiotic (for example an anthracycline sue adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-' dactinomycin or mithramycin); an antimitotic agent (for example a vinca alkaloid sucl

topoisomerase inhibitor (for example an epipodophyllotoxin such as etoposide, tenipo 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 exai 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 inhibitc 5α-reductase such as finasteride; (iii) an agent which inhibits cancer cell invasion (for example a metalloproteinase inhi 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/threonii kinase inhibitor, an inhibitor of the epidermal growth factor family (for example an E( 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)-( bis(2-methoxyethoxy)quinazolin-4-amine (erlotinib, OSI-774) or 6-acrylamido-N-(3- chloro-4-fluorophenyl)-7-(3-moφholinopropoxy)quinazolin-4-a mine (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 endothel 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 < WO 98/13354), or a compound that works by another mechanism (for example linomi an inhibitor of integrin αvβ3 function or an angiostatin);

(vi) a vascular damaging agent such as combretastatin A4, or a compound disclosed in 99/02166, WO 00/40529, WO 00/41669, WO 01/92224, WO 02/04434 or WO 02/082 (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 abe

pro-drug therapy) approaches such as those using cytosine deaminase, thymidine kim 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-viv approaches to increase the immunogenicity of patient tumour cells, such as transfectic with cytokines such as interleukin 2, interleukin 4 or granulocyte-macrophage colony stimulating factor, approaches to decrease T-cell anergy, approaches using transfectec 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 prese invention, as hereinbefore described, and at least one further active ingredient selectee 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.

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 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 respira) diseases such as asthma, COPD or rhinitis.

Examples of a phosphodiesterase inhibitor that may be used in the pharmaceutical pro according to this embodiment include a PDE4 inhibitor such as an inhibitor of the isof

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

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

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

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

N-(3,5,dichloro-4-pyridinyl)-2-[l-(4-fluorobenzyl)-5-hydr oxy-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-j k][l,4]benzodiazepin-3( yl]pyridine-4-carboxamide (CI-1018), cis-[4-cyano-4-(3-cyclopentyloxy-4-methoxyphenyl)cyclohexane -l-carboxylic acid

(Cilomilast)

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

5-(3,5-di-tert-butyl-4-hydroxybenzylidene)-2-iminothiazol idin-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 (Lirimila

(-)-(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][l,6]naphthyridine (Pumafentrine),

3-(cyclopropylmethoxy)-N-(3,5-dichloro-4-pyridyl)-4-(difl uoromethoxy)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-meth yl-4H-pyrimido[6,l-

3-[[3-(cyclopentyloxy)-4-methoxyphenyl]-methyl]-N-ethyl-8 -(l-methylethyl)-3H-pur 6-amine (V-11294A).

Examples of a β ₂ -adrenoceptor agonist that may be used in the pharmaceutical produc according to this embodiment include metaproterenol, isoproterenol, isoprenaline, albuterol, salbutamol (e.g. as sulphate), formoterol (e.g. as fumarate), salmeterol (e.g. xinafoate), terbutaline, orciprenaline, bitolterol (e.g. as mesylate), pirbuterol or indaca The β ₂ -adrenoceptor agonist of this embodiment may be a long-acting β ₂ -agonists, for example salmeterol (e.g. as xinafoate), formoterol (e.g. as fumarate), bambuterol (e.g. 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 Registi Number 137888-11-0 and disclosed in U.S. Patent No 4,579,854), indacaterol (CAS n 312753-06-3; QAB-149), formanilide derivatives e.g. 3-(4-{ [6-({ (2R)-2-[3- (formylamino)-4-hydroxyphenyl]-2-hydroxyethyl}amino)hexyl]ox y}-butyl)- benzenesulfonamide as disclosed in WO 2002/76933, benzenesulfonamide derivatives 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 and US 2005/222144, and compounds G 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 antagc

Examples of an inhibitor of kinase function that may be used in the pharmaceutical pr< 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 accoi to this embodiment include an inhibitor of neutrophil elastase or an inhibitor of MMPl

Examples of a steroidal glucocorticoid receptor agonist that may be used in the pharmaceutical product according to this embodiment include budesonide, fluticasom 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, zoticas< flumoxonide, rofleponide, butixocort (e.g. as propionate ester), prednisolone, prednisc tipredane, steroid esters e.g. 6α,9α-difluoro-17α-[(2-furanylcarbonyl)oxy]-l lβ-hydrc 16α-methyl-3-oxo-androsta-l,4-diene-17β-carbothioic acid S-fluoromethyl ester, 6α, 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-difluoro-l lβ-hydn 16α-methyl- 17α-[(4-methyl- 1 ,3-thiazole-5-carbonyl)oxy]-3-oxo-androsta- 1 ,4-diene- ] carbothioic acid S -fluoro methyl ester, steroid esters according to DE 4129535, steroic according to WO 2002/00679, WO 2005/041980, or steroids GSK 870086, GSK 685( and GSK 799943.

Examples of a modulator of a non-steroidal glucocorticoid receptor agonist that may 1 used in the pharmaceutical product according to this embodiment include those descri in WO2006/046916.

The invention is illustrated by the following Examples. In the Examples the followinj Figures are presented:

Figure 1: X-ray powder diffraction pattern of Form A of Example 14. Figure 2: X-ray powder diffraction pattern of Form A of Example 15.

In the examples the NMR spectra were measured on a Varian Unity Inova spectromet a proton frequency of either 300 or 400 or 500 MHz, or on a Bruker DRX spectrometi a proton frequency of 400 or 500 MHz, or on a Bruker Avance spectrometer with a pr frequency of 600 MHz or or on a Bruker Avance DPX 300 spectrometer with a protor frequency of 300 MHz. The MS spectra were measured on either an Agilent 1100 MS G1946D spectrometer or a Hewlett Packard HPl 100 MSD G1946A spectrometer or a

Waters Micromass ZQ2000 spectrometer. Names were generated using the Autonom (version 4.01.305) software supplied by MDL.

XRPD data were collected using either a PANalytical CubiX PRO machine or a PANalytical X-Pert machine.

XRPD - PANalvtical CubiX PRO

Data was collected with a PANalytical CubiX PRO machine in θ- # configuration ovi scan range 2° to 40° 2 θ with 100-second exposure per 0.02° increment. The X-rays w generated by a copper long-fine focus tube operated at 45kV and 4OmA. The waveler of the copper X-rays was 1.5418 A . The Data was collected on zero background hole on which ~ 2 mg of the compound was placed. The holder was made from a single cr of silicon, which had been cut along a non-diffracting plane and then polished on an optically flat finish. The X-rays incident upon this surface were negated by Bragg extinction.

PANalvtical X-Pert

Data was collected using a PANalytical X-Pert machine in 2θ - # configuration over I scan range 2° to 40° 2#with 100-second exposure per 0.02° increment. The X-rays w generated by a copper long-fine focus tube operated at 45kV and 4OmA. The waveler of the copper X-rays was 1.5418A . The Data was collected on zero background hold on which ~ 2 mg of the compound was placed. The holder was made from a single cr of silicon, which had been cut along a non-diffracting plane and then polished on an optically flat finish. The X-rays incident upon this surface were negated by Bragg extinction.

DSC thermograms were measured using a TA QlOOO Differential Scanning Calorimei with aluminium pans and pierced lids. The sample weights varied between 0.5 to 5 m The procedure was carried out under a flow of nitrogen gas (50 mL/min) and the temperature studied from 25 to 300 ⁰ C at a constant rate of temperature increase of 10 ^c per minute.

TGA thermograms were measured using a TA Q500 Thermogravimetric Analyser, wi platinum pans. The sample weights varied between 1 and 5 mg. The procedure was carried out under a flow of nitrogen gas (60 mL/min) and the temperature studied fron to 300 ⁰ C at a constant rate of temperature increase of 1O ⁰ C per minute.

GVS profiles were measured using a Dynamic Vapour Sorption DVS-I instrument. T solid sample ca. 1-5 mg was placed into a glass vessel and the weight of the sample w, recorded during a dual cycle step method (40 to 90 to 0 to 90 to 0% relative humidity (RH), in steps of 10% RH).

Abbreviations used in the experimental section:

Aq = aqueous

DCE = 1 ,2-dichloroethane

DCM = dichloromethane DMF = dimethylformamide

DMSO = Dimethylsulfoxide

EtOAc = ethyl acetate

EtOH = ethanol

GVS = Gravimetric vapour sorption HATU = 0-(7-Azabenzotriazol-l-yl)-N,N,/V',./V'-tetramethyluronium hexafluorophosp

MeCN - Acetonitrile

MeOH = methanol

RT = RT

Rt = retention time THF = tetrahydrofuran

Satd = saturated

DSC = Differential Scanning Calorimetry

TGA = Thermogravimetric analysis

XRPD = X-Ray Powder Diffraction

Example 1: (R)-l-[(6-Methyl-pyridin-3-ylcarbamoyl)-methyI]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride a) 1-Phenyl-cycloheptanol

To magnesium (1.2 g) in anhydrous tetrahydrofuran (60 mL) under an environment oi nitrogen was added a crystal of iodine followed by bromobenzene (7.85 g) at such a π that the reaction maintained a steady reflux. The reaction mixture was stirred for 20 minutes then cycloheptanone (4.48 g) was added with care. After stirring for 10 minu saturated aqueous ammonium chloride (10 mL) was added and the reaction was partit between water (100 mL) and isohexane (100 mL). The organic layer was dried (MgS ¹ and evaporated to afford the sub-titled compound (7.6 g) as an oil.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.53 - 7.47 (m, 2H), 7.36 - 7.29 (m, 2H), 7.26 - 7.19 ( IH), 2.07 (ddd, 2H), 1.97 - 1.50 (m, 1 IH).

b) 1 -Methoxy- 1 -phenyl-cycloheptane

1-Phenyl-cycloheptanol (Example Ia) (7.6 g) was dissolved in tetrahydrofuran (100 rr and sodium hydride (60% in oil, 2.0 g) added. The reaction was stirred at 60°C for 5 minutes and iodomethane (7.1 g) added. The mixture was maintained at 60 ⁰ C overni^ and then further quantities of sodium hydride (60% in oil, 2.0 g) and iodomethane (7.1 were added and the reaction was refluxed for 70 hours. The reaction mixture was f 1 DO ml ^ QnH th^ l αλrøt-

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.43 - 7.37 (m, 2H), 7.37 - 7.30 (m, 2H), 7.24 - 7.19 ( IH), 2.98 (s, 3H), 2.12 - 1.88 (m, 4H), 1.88 - 1.45 (m, 8H).

c) 1-Phenyl-cycloheptanecarboxylic acid

Potassium (2.62 g) and sodium (0.52 g) were heated together at 120 ⁰ C in mineral oil i an environment of nitrogen for 30 minutes and then cooled to room temperature. The < was removed and replaced with ether (100 mL) and l-methoxy-l-phenyl-cycloheptan< (Example Ib) (4.9 g) was added and the reaction was stirred under nitrogen overnight room temperature. The reaction was cooled to -78°C and solid carbon dioxide (-20 g^ added with stirring. The reaction was allowed to warm to room temperature and watei (150 mL) was added carefully under an environment of nitrogen. The aqueous layer v separated, neutralised with concentrated hydrochloric acid and extracted with diethyl c (150 mL). The organic layer was dried (MgSO ₄ ) and evaporated afford to the sub-title compound (4.15 g) as an oil.

¹ H NMR (300 MHz, CDCl ₃ ) 6 7.40 - 7.20 (m, 5H), 2.49 - 2.35 (m, 2H), 2.16 - 2.03 ( 2H), 1.76 - 1.47 (m, 8H).

d) 1 -Phenyl -cycloheptanecarboxylic acid methyl ester

1-Phenyl-cycloheptanecarboxylic acid (Example Ic) (4.15 g) was refluxed in methane (150 mL) and concentrated hydrochloric acid (5 mL) for 24 hours. The solvent was evaporated and the residue was dissolved in ether (100 mL) which was washed with v (100 mL), saturated sodium bicarbonate (50 mL) and water (100 mL), dried (MgSO ₄ ) evaporated to afford the sub-titled compound (3.5 g) as an oil.

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.37 - 7.18 (m, 5H), 3.63 (s, 3H), 2.47 - 2.35 (m, 2H^ 2.08 - 1.97 (m, 2H), 1.70 - 1.48 (m, 8H).

e) 1-Phenyl-cycloheptanecarboxylic acid (./?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester

H

1-Phenyl-cycloheptanecarboxylic acid methyl ester (Example Id) (1.0 g) and (R)- quinuclidin-3-ol (0.39 g) were refluxed in heptane (50 mL) containing sodium (~5 mg Dean and Stark apparatus for 24 hours. Heptane (20 mL) was replaced with toluene ('. mL) and the reflux was continued for 3 days. The reaction was partitioned between w (50 mL) and ether (50 mL) and the ether layer was separated, dried (MgSO ₄ ) and evaporated. The crude product was purified by column chromatography on silica elut with ethyl acetate / triethylamine (99/1) to afford the titled compound as an oil (0.83 g

m/e 328 [M+H] ⁺

¹ H NMR (300 MHz, CDCl ₃ ) 6 7.35 - 7.27 (m, 4H), 7.23 - 7.16 (m, IH), 4.78 - 4.71 ( IH), 3.12 (ddd, IH), 2.79 - 2.32 (m, 7H), 2.16 - 1.98 (m, 2H), 1.91 - 1.80 (m, IH), 1 1.34 (m, 12H).

f) 2-Chloro-./V-(6-methyl-pyridin-3-yl)-acetamide

A mixture of 6-methylpyridin-3-amine (Ig) and triethylamine (2.2 mL) in dry THF (2 mL) was stirred and cooled to -6O ⁰ C. 2-Chloroacetyl chloride (1.567 g) was added via syringe to the stirred mixture forming a yellow suspension. The mixture was stirred at 60 ⁰ C until analysis showed complete dissappearance of starting material. The reactior slurry was poured into water and the products extracted with ethyl acetate (2 x 150 ml The combined organic extracts were dried over magnesium sulphate and concentrated dryness. The crude brown solid was recrystallised from ether to afford the subtitled compound (700 mg).

¹ H NMR (400 MHz, DMSO-D ₆ ) 5 10.40 (IH, s), 8.60 (IH, d), 7.91 (IH, dd), 7.22 (IF 4.27 (2H, s), 2.42 (3H, s).

Example 1: (/?)-l-[(6-MethyI-pyridin-3-ylcarbamoyl)-methyI]-3-(l-phenyl - cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

H

1-Phenyl-cycloheptanecarboxylic acid (/?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Exam] Ie) (52 mg) was dissolved in acetonitrile (2 mL) and 2-chloro-N-(6-methylpyridin-3- yl)acetamide (Example If) (29 mg) was added. The reaction mixture was stirred for 11 days and diluted with ethyl acetate (4 mL) and /sσhexane (14 mL). The mixture was 1 standing for 5 days, whereupon the resulting crystals were separated and washed with

m/e 476 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 1 1.33 (s, IH), 8.70 (d, IH), 7.91 (dd, IH), 7.38 - ' (m, 4H), 7.27 (d, IH), 7.28 - 7.20 (m, IH), 5.16 - 5.07 (m, IH), 4.36 (d, IH), 4.31 (c s IH), 4.16 - 4.07 (m, IH), 3.72 - 3.54 (m, 4H), 3.44 - 3.34 (m, 2H), 2.44 (s, 3H), 2.4 2.28 (m, 2H), 2.22 - 2.10 (m, 2H), 2.01 - 1.86 (m, 3H), 1.83 - 1.71 (m, IH), 1.69 - 1 (m, 8H).

Example 2: (/?)-l-[(6-Methyl-pyrazin-2-ylcarbamoyl)-methyl]-3-(l-phenyl -o cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-./V-(6-methyl-pyrazin-2-yl)-acetamide

s 6-Methyl-pyrazin-2-ylamine (150 mg) and potassium carbonate (571 mg) were added dichloromethane (25 niL). 2-Bromoacetyl bromide (0.120 mL) was added to the suspension with stirring. The reaction was stirred overnight then water (0.1 mL) was added with further stirring. Further quantities of potassium carbonate (571 mg), 2- bromoacetyl bromide (0.120 mL) and water (0.1 mL) were added over 2 hours until tr reaction had proceeded to completion. The reaction was diluted with water (100 mL), carefully acidified with hydrochloric acid and extracted with dichloromethane (2 x 50 which was dried and evaporated to afford the sub-titled compound which was used cπ (365 mg).

1H NMR (400 MHz, CDCl ₃ ) 59.45 (s, IH), 9.37 (s, IH), 8.33 (s, IH), 4.05 (s, 2H), 2. 3H).

Example 2: (R)-l-[(6-Methyl-pyrazin-2-ylcarbamoyI)-methyl]-3-(l-phenyl-

H

1-Phenyl-cycloheptanecarboxylic acid (R)-(I -aza-bicyclo[2.2.2]oct-3-yl) ester (Examj Ie) (70 mg) and 2-bromo-N-(6-methylpyrazin-2-yl)acetamide (Example 2a) (49.2 mg; were dissolved in acetonitrile (1 niL) and left to stand overnight. Crystals separated o standing and were filtered and washed with acetonitrile (2 x 1 mL), ethyl acetate (2 x : mL) and diethyl ether (2 x 3 mL) and dried to yield the titled compound (24 mg).

m/e 477 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 11.33 (s, IH), 9.09 (s, IH), 8.37 (s, IH), 7.38 - 7. (m, 4H), 7.27 - 7.22 (m, IH), 5.15 - 5.09 (m, IH), 4.36 - 4.25 (m, 2H), 4.16 - 4.07 (i IH), 3.68 - 3.56 (m, 4H), 3.46 - 3.33 (m, IH), 2.47 (s, 3H), 2.42 - 2.29 (m, 2H), 2.2 _* 2.1 1 (m, 2H), 2.04 - 1.87 (m, 3H), 1.83 - 1.73 (m, IH), 1.68 - 1.45 (m, 9H).

Example 3: (/?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-[(6-trifluoromet hyI- pyridazin-3-ylcarbamoyl)-methyl]-l-azonia-bicyclo[2.2.2]octa ne bromide

a) 2-Bromo-N-(6-trifluoromethyl-pyridazin-3-yl)-acetamide

6-Trifluoromethyl-pyridazin-3-ylamine (0.042 g) (prepared by a procedure similar to t described in WO2007048779) was dissolved in dichloromethane (40 mL) and stirred \ potassium carbonate (0.214 g). 2-Bromoacetyl bromide (0.12 mL) was added and stiri continued for 1.5 hours. Water (0.24 mL) was added and the reaction mixture was stin fnr 1 5 hnπrς affp.r whirh water (AO ml λ wac λλλPλ anri fhp rpnrtinn miγtiirp ctirrpri fnr

further 1.5 hours. The dichloromethane was separated, dried (MgSO ₄ ) and evaporated afford the sub-titled compound as a white solid (0.053 g).

m/e 284/286 [M+H] ⁺ 1H NMR (400 MHz, DMSO-D ₆ ) δ 11.04 (s, IH), 8.79 (d, IH), 7.91 (d, IH), 4.31 (s,

Example 3: (/f)-3-(l-Phenyl-cycloheptaπecarbonyloxy)-l-[(6-trifluorome thyl- pyridazin-3-ylcarbamoyl)-methyl]-l-azonia-bicyclo[2.2.2]octa ne bromide

1-Phenyl-cycloheptanecarboxylic acid (./?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Examj Ie) (61.1 mg) and 2-bromo-/V-(6-trifluoromethyl-pyridazin-3-yl)-acetamide (Example (53.0 mg) were dissolved in acetonitrile (2 mL) and left overnight. The solvent was evaporated and the product was purified by column chromatography on silica eluting > 10% methanol in dichloromethane to afford the titled compound (107 mg).

m/e 531 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 12.17 (s, IH), 8.50 (d, IH), 8.36 (d, IH), 7.40 - 7.2 (m, 4H), 7.28 - 7.23 (m, IH), 5.17 - 5.10 (m, IH), 4.57 - 4.42 (m, 2H), 4.22 - 4.14 (i IH), 3.76 - 3.61 (m, 4H), 3.47 (dd, IH), 2.43 - 2.30 (m, 2H), 2.25 - 2.12 (m, 2H), 2.C 1.88 (m, 3H), 1.86 - 1.73 (m, IH), 1.72 - 1.44 (m, 9H).

Example 4: (R)-l-(Benzo[rf]isoxazol-3-yIcarbamoylmethyl)-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

To a mixture of benzo[<i]isoxazol-3-ylamine (1 g) and cesium carbonate (2.42 g) in dr DMF (20 mL), stirred at it, was added bromoacetyl chloride (0.62 mL) by dropwise addition. After stirring the mixture for δhours, the reaction was poured into water (IOC and the products extracted into ether (2 x 200 mL). The combined extracts were dried magnesium sulfate and concentrated to dryness. The crude product was purified on sil gel using ether / isohexane, (4 / 6) to afford the sub-titled compound as a colourless sol (0.5 g).

m/e 210 [M+H] ⁺

Example 4: (i?)-l-(Benzo[J]isoxazol-3-ylcarbamoylmethyl)-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

H

1 -Phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Examr. Ie) (1 14 mg) and /V-benzo[ύT|isoxazol-3-yl-2-chloro-acetamide (Example 4a) (89 mg) dissolved in acetonitrile (10 mL) and left for one week. The resulting crystals were filt off and washed with diethyl ether (3 x 10 mL) to afford the titled compound as a solid (120 mg).

m/e 502 [M] ⁺

¹ H NMR (ά(Y) MH ₇ F)MSO-D^ λ 1 ? 1 S CQ 1 Hϊ 8 I f> (ή λ U\ 1 IA (A 1 Hϊ 7 10 . 1 λ

4.63 - 4.46 (m, 2H), 4.17 (ddd, IH), 3.76 - 3.61 (m, 4H), 3.49 (dd, IH), 2.43 - 2.29 (m 2H), 2.24 - 2.12 (m, 2H), 2.03 - 1.89 (m, 3H), 1.86 - 1.74 (m, IH), 1.70 - 1.44 (m, 9H)

Example 5: (/f)-l-(Pyridazin-3-ylcarbamoylmethyl)-3-(l-thiophen-2-yl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-But-3-enyl-2-thiophen-2-yl-hex-5-enoic acid ethyl ester

Ethyl 2-(thiophen-2-yl)acetate (2.35 g) was dissolved in tetrahydrofuran (30 mL) and cooled to -78°C. Lithium Z?w(trimethylsilyl)amide (2.31 g) in THF (IM solution, 13.8 was added and the solution was stirred for 30 minutes. 4-Bromo-but-l-ene (1.4 mL) w added and the reaction mixture was allowed to warm to room temperature and stirred f hour. The reaction mixture was re-cooled to -78°C and lithium b/s(trimethylsilyl)amic (2.31 g) in THF (IM solution, 13.8 mL) was added and the solution was stirred for 30 minutes. 4-Bromo-but-l-ene (1.4 mL) was added and the reaction mixture was allowe warm to room temperature and stand overnight. HPLC-MS analysis indicated that the reaction was incomplete so the reaction was again cooled to -78°C and further aliquots lithium bw(trimethylsilyl)amide (IM solution, 10 mL) and 4-bromo-but-l-ene (1.0 mL were added following the procedure outlined above. After stirring for a further 2 hours water (30 mL) was added and the reaction was extracted with diethyl ether (2 x 60 mL The combined organic extracts were dried (MgSO ₄ ) and evaporated. The resulting oil ' purified by column chromatography on silica eluting with ethyl acetate / /søhexane (1/ to afford the sub-titled compound (3.18 g).

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 7.21 (dd, IH), 6.97 - 6.94 (m, 2H), 5.79 (ddt, 2H), (dq, 2H), 4.95 (dq, 2H), 4.17 (q, 2H), 2.22 - 2.08 (m, 4H), 2.00 - 1.85 (m, 4H), 1.24 (I αi_τ\

b) l-Thiophen-2-yl-cyclohept-4-enecarboxylic acid ethyl ester

To 2-but-3-enyl-2-thiophen-2-yl-hex-5-enoic acid ethyl ester (Example 5a) (3.18 g) in dichloromethane (100 mL) was added Grubbs Catalyst (2nd Generation, Sigma- Aldrii Company Ltd) (0.100 g). The mixture was warmed to reflux under nitrogen. After 20 the mixture was allowed to cool to room temperature and evaporated to an oil. Purific by column chromatography on silica eluting with ethyl acetate / wohexane (10:90) to ; the sub-titled compound (2.60 g) as a coloured oil.

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 7.19 (dd, IH), 6.98 - 6.92 (m, 2H), 5.72 (t, 2H), 4.1 2H), 2.66 - 2.59 (m, 2H), 2.25 - 2.14 (m, 6H), 1.21 (t, 3H).

c) l-Thiophen-2-yl-cycloheptanecarboxylic acid ethyl ester

l-Thiophen-2-yl-cyclohept-4-enecarboxylic acid ethyl ester (Example 5b) (2.86 g) wa dissolved in ethanol (30 mL) and tris(triphenylphosphine)rhodium(I) chloride (0.100 % was added. The reaction mixture was stirred rapidly under 5 atmospheres of hydrogen overnight. Further tris(triphenylphosphine)rhodium(I) chloride (0.050 g) was added a the reaction mixture was stirred under 5 atmospheres of hydrogen for 3 days. A third addition of tris(triphenylphosphine)rhodium(I) chloride (0.050 g) was made and the reaction mixture was stirred under 3 atmospheres of hydrogen overnight. The content were evaporated to dryness and purified on silica eluting with ethyl acetate / isohexam

m/e 253 [M+H ⁺ ]

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 7.17 (dd, IH), 6.95 - 6.91 (m, 2H), 4.13 (q, 2H), 2

(dd, 2H), 2.14 - 2.03 (m, 2H), 1.70 - 1.50 (m, 8H), 1.20 (t, 3H).

d) l-Thiophen-2-yl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) esi

H l-Thiophen-2-yl-cycloheptanecarboxylic acid ethyl ester (Example 5c) (2.5 g) and (R) quinuclidin-3-ol (2.08 g) were dissolved in toluene (350 mL) and sodium hydride (0.1 added under nitrogen. The mixture was heated to reflux for 20 hours after which the toluene was carefully distilled off to leave -100 mL which was cooled and washed wi' water (100 mL), dried (MgSO ₄ ) and evaporated. The crude product was purified by column chromatography on silica eluting with ethyl acetate / triethylamine (99/1) to a) the sub-titled compound (2.84 g).

m/e 334 [M+H ⁺ ]

¹ H NMR (400 MHz, CDCl ₃ ) 5 7.18 (t, IH), 6.95 - 6.92 (m, 2H), 4.77 - 4.72 (m, IH), (ddd, IH), 2.83 - 2.64 (m, 4H), 2.59 - 2.50 (m, 3H), 2.18 - 2.08 (m, 2H), 1.95 - 1.90 (π IH), 1.71 - 1.44 (m, 11H), 1.34 - 1.23 (m, IH).

e) 2-Bromo-/V-pyridazin-3-yl-acetamide

To a suspension of pyridazin-3-ylamine (2.7 g) and di/sopropylethylamine (6.3 mL) ii dichloromethane (100 mL) at 0 ⁰ C was added bromoacetic anhydride (9.0 g) in dichloromethane (10 mL) by dropwise addition. The mixture was stirred at O ⁰ C for 0. hours and then allowed to warm to rt. The resulting suspension was filtered, washed v dichloromethane and dried to afford the sub-titled compound as a solid (2.0 g).

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 1 1.51 (s, IH), 9.00 (dd, IH), 8.28 (dd, IH), 7.74 - ' (m, IH), 4.15 (s, 2H).

Example 5: (R)-l-(Pyridazin-3-ylcarbamoyImethyl)-3-(l-thiophen-2-yI- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

l-Thiophen-2-yl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 5d) (80 mg) and 2-bromo-./V-pyridazin-3-yl-acetamide (Example 5e) (52 mg were dissolved in acetonitrile (3 mL) and stirred overnight. Ethyl acetate (9 mL) and isohexane (4 mL) were added and stirred overnight. The resulting crystals were filtere and then triturated with ethyl acetate to afford the titled compound (14 mg).

m/e 469 [M ⁺ ]

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 1 1.68 (s, IH), 9.05 (dd, IH), 8.25 (d, IH), 7.79 (dc IH), 7.44 (dd, IH), 7.03 (dd, IH), 6.99 (dd, IH), 5.14 - 5.09 (m, IH), 4.37 (s, 2H), 4.1 4.08 (m, IH), 3.76 - 3.57 (m, 4H), 3.57 - 3.46 (m, IH), 2.48 - 2.42 (m, IH), 2.29 - 2.2S IH), 2.21 - 2.1 1 (m, IH), 2.07 - 1.90 (m, 4H), 1.90 - 1.80 (m, IH), 1.78 - 1.68 (m, IH) 1.66 - 1.46 (m, 8H).

a) 2-Bromo-iV-(5-methyl-isoxazol-3-yl)-acetamide

To a stirred suspension of sodium bicarbonate (1.242 g) and 5-methyl-isoxazol-3-ylaπ (1.45 g) in dichloromethane (50 mL) was added 2-bromoacetyl bromide (1.28 mL) by dropwise addition. The reaction mixture was stirred overnight and then washed with v (2 x 50 mL). The organic fraction was separated, dried with magnesium sulfate and evaporated to yield the sub-titled compound (279 mg).

¹ H NMR (400 MHz, DMSO-D ₆ ) 5 11.32 (s, IH), 6.62 (s, IH), 4.06 (s, 2H), 2.38 (s,

Example 6: (R)-l-[(5-MethyI-isoxazol-3-ylcarbamoyl)-methyl]-3-(l-thioph en-2-yl cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

l-Thiophen-2-yl-cycloheptanecarboxylic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 5d) (68 mg) and 2-bromo-./V-(5-methyl-isoxazol-3-yl)-acetamide (Example I (45 mg) were dissolved in acetonitrile (2 mL) and stirred overnight. Ethyl acetate (10 i and isohexane (9 mL) were added and stirred overnight. The resulting crystals were filtered off and washed with ethyl acetate to afford the titled compound (82 mg).

m/e 472 [M ⁺ ]

- 3.55 (m, 4H), 3.53 - 3.43 (m, IH), 2.54 - 2.42 (m, IH), 2.41 (d, 3H), 2.27 - 2.22 (m, 2.19 - 2.13 (m, IH), 2.07 - 1.90 (m, 4H), 1.89 - 1.77 (m, IH), 1.77 - 1.65 (m, IH), 1.6: 1.48 (m, 8H).

Example 7: (λ)-l-[(3-Methyl-isoxazol-5-ylcarbamoyI)-methyl]-3-(l-thiop hen-2-yl cycloheptanecarbonyloxy)- 1 -azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-./V-(3-methyl-isoxazol-5-yl)-acetamide

3-Methyl-isoxazol-5-ylamine (2.9 g) and potassium carbonate (9.8 g) were suspended dichloromethane (100 mL) at room temperature and 2-bromoacetyl bromide (6 g) was added dropwise. The mixture was allowed to stir overnight. Water (0.3 mL) was addec together with a further quantity of potassium carbonate (3 g) and the reaction mixture stirred for a further 30 minutes. The reaction mixture was poured into water (100 mL) extracted with dichloromethane (2 x 50 mL). The combined organic extracts were drie over magnesium sulfate and then evaporated in vacuo. The crude product was purifed column chromatography on silica eluting with ethyl actetate / isohexane (50:50) to gή sub-titled compound (4.8 g).

¹ H NMR (300 MHz, CDCl ₃ ) 5 1 1.97 (s, IH), 6.16 (s, IH), 4.09 (s, 2H), 2.19 (s, 3H)

Example 7: (R)-l-[(3-Methyl-isoxazoI-5-ylcarbamoyl)-methyl]-3-(l-thioph en-2-yI- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

l-Thiophen-2-yl-cycloheptanecarboxylic acid (/?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 5d) (50 mg) and 2-bromo-N-(3-methylisoxazol-5-yl)acetamide (Example 7a mg) were dissolved in acetonitrile (2 mL) and left overnight. Ethyl acetate (10 mL) an /søhexane (10 mL) were added and the crystals filtered off, washed with ethyl acetate dried to afford the titled compound (37 mg).

m/e 472 [M ⁺ ] io ¹ H NMR (400 MHz, DMSO-D ₆ ) δ 12.21 (s, IH), 7.44 (dd, IH), 7.03 (dd, IH), 6.99 (c

IH), 6.18 (s, IH), 5.15 - 5.07 (m, IH), 4.35 (d, IH), 4.30 (d, IH), 4.14 - 4.05 (m, IH),

- 3.54 (m, 4H), 3.54 - 3.43 (m, IH), 3.17 (d, IH), 2.47 - 2.42 (m, IH), 2.27 - 2.20 (m,

2.21 (s, 3H), 2.19 - 2.12 (m, IH), 2.08 - 1.77 (m, 4H), 1.77 - 1.65 (m, IH), 1.65 - 1.46

8H).

I ₅

Example 8: (R)-l-[(3-Fluoro-phenylcarbamoyl)-methyl]-3-(l-thiophen-2-yl - cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-/V-(3-fluoro-phenyl)-acetamide

20

To a suspension of sodium bicarbonate (1 g) and 3-fluoroaniline (0.46 g) in dichloromethane (100 mL) was added 2-bromoacetyl bromide (0.36 mL) by dropwise

addition. After stirring overnight the reaction mixture was washed with water, dried w magnesium sulfate and evaporated to yield the sub-titled compound (1.07 g).

m/e 232 [M+H ⁺ ] s ¹ H NMR (300 MHz, CDCl ₃ ) δ 8.14 (s, IH), 7.50 (dt, IH), 7.31 (td, IH), 7.18 (ddd, 11 6.87 (tdd, IH), 4.03 (s, 2H).

Example 8: (/?)-l-[(3-FIuoro-phenylcarbamoyl)-methyl]-3-(l-thiophen-2-y l- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide 0

l-Thiophen-2-yl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 5d) (96 mg) and 2-bromo-./V-(3-fluoro-phenyl)-acetamide (Example 8a) (67 were dissolved in acetonitrile (2 mL) and left overnight. Diethyl ether (10 mL) andS /sohexane (8 mL) were added and the mixture was left overnight. The resulting crysta were filtered off and washed with diethyl ether to afford the titled compound (90 mg).

m/e 485 [M ⁺ ]

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 10.84 (s, IH), 7.58 (dt, IH), 7.46 - 7.39 (m, 2H), 7.0 7.29 (m, IH), 7.04 (dd, IH), 7.02 - 6.96 (m, 2H), 5.15 - 5.10 (m, IH), 4.33 - 4.24 (m, 4.17 - 4.07 (m, IH), 3.77 - 3.58 (m, 4H), 3.51 (dd, IH), 2.56 - 2.44 (m, IH), 2.28 - 2.2 IH), 2.21 - 2.12 (m, IH), 2.08 - 1.70 (m, 6H), 1.66 - 1.47 (m, 8H).

Example 9: (/?)-l-[(5-Methyl-pyrazin-2-yIcarbamoyl)-methyl]-3-(l-thioph en-2-yl5 cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

To a mixture of 5-methyl-pyrazin-2-ylamine and cesium carbonate (11.2 g) dissolved dry DMF (30 mL) was added by dropwise addition bromoacetylbromide (2.89 g) and mixture stirred at rt for 2 hours. Water (200 mL) was added and the mixture extracted ethyl acetate (2 x 100 mL) and dried over magnesium sulfate. Concentration of the exi to -50 mL and addition of tsøhexane (100 mL) gave the sub-titled compound as a sol (1-64 g).

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 1 1.06 (IH, s), 9.17 (IH, s), 8.31 (IH, d), 4.16 (2H, 2.46 (3H, s).

Example 9: (/f)-l-[(5-Methyl-pyrazin-2-ylcarbamoyl)-methyl]-3-(l-thioph en-2-yl- cycloheptanecarbonyloxy)- 1 -azonia-bicyclo[2.2.2]octane bromide

l-Thiophen-2-yl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 5d) (48 mg) was dissolved in acetonitrile (2 mL) and 2-bromo-N-(5-methyl- pyrazin-2-yl)-acetamide (Example 9a) (33 mg) was added. After stirring for 1 week diethyl ether (8 mL) and wøhexane (5 mL) were added. The crystals were collected b] filtration, washed with ethyl acetate (2 x 4 mL) and dried to afford the titled compounc mg).

¹ H NMR (400 MHz, DMSO-D ₆ ) 6 11.26 (s, IH), 9.15 (s, IH), 8.36 (s, IH), 7.44 (dd, 7.04 (dd, IH), 6.99 (dd, IH), 5.15 - 5.08 (m, IH), 4.33 (s, 2H), 4.13 (ddd, IH), 3.75 - : (m, 4H), 3.56 - 3.46 (m, IH), 2.48 (s, 3H), 2.50 - 2.44 (m, IH), 2.28 - 2.22 (m, IH), 2. 2.11 (m, IH), 2.08 - 1.90 (m, 4H), 1.90 - 1.80 (m, IH), 1.79 - 1.69 (m, IH), 1.64 - 1.4! 8H).

Example 10: (/?)-l-(Benzo[rf]isoxazol-3-ylcarbamoylmethyl)-3-(l-thiophen -2-yl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

l-Thiophen-2-yl-cycloheptanecarboxylic acid (λ)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 5d) (71 mg) and N-benzo[eT|isoxazol-3-yl-2-chloro-acetamide (Example 4a) mg) were dissolved in acetonitrile (10 mL) and left to stand for 6 days. The resulting crystals were filtered off and washed with diethyl ether (3 x 10 mL) to afford the titled compound (82 mg).

m/e 509 [M ⁺ ]

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 12.16 (s, IH), 8.17 (d, IH), 7.74 (d, IH), 7.72 - 7.6 IH), 7.44 - 7.39 (m, 2H), 7.04 (dd, IH), 6.98 (dd, IH), 5.16 - 5.1 1 (m, IH), 4.64 - 4.5C 2H), 4.21 - 4.13 (m, IH), 3.82 - 3.64 (m, 4H), 3.59 (dd, IH), 2.56 - 2.44 (m, 2H), 2.29 2.22 (m, IH), 2.22 - 2.13 (m, IH), 2.08 - 1.89 (m, 3H), 1.89 - 1.81 (m, IH), 1.80 - 1.6S IH), 1.64 - 1.47 (m, 8H).

Example 11: (R)-l-(Pyrazin-2-yIcarbamoylmethyl)-3-(l-thiophen-2-yl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

To a stirred suspension of pyrazin-2-ylamine (1.87 g) and potassium carbonate (8.19 j dichloromethane (25 mL) was added by dropwise addition 2-bromoacetyl bromide (1, mL). The reaction mixture was stirred overnight and then washed with water (2 x 50 i The organic phase was separated, dried with magnesium sulfate and evaporated to yie sub-titled compound (0.70 g).

¹ H NMR (400 MHz, CDCl ₃ ) δ 9.51 (d, IH), 8.63 (s, IH), 8.42 (d, IH), 8.30 (dd, IH), (s, 2H).

Example 11: (/?)-l-(Pyrazin-2-ylcarbamoylmethyl)-3-(l-thiophen-2-yl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

H l-Thiophen-2-yl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 5d) (116 mg) and 2-bromo-/V-pyrazin-2-yl-acetamide (Example 1 Ia) (75 mj were dissolved in acetonitrile (2 mL) and left overnight. Diethyl ether (10 mL) and /.røhexane (8 mL) were added and the mixture was left to stand overnight. The resulti: crystals were filtered off and washed with diethyl ether to afford the titled compound I mg).

m/e 469 [M ⁺ ]

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 1 1.38 (s, IH), 9.28 (s, IH), 8.50 - 8.45 (m, 2H), IA

(dd, IH), 7.04 (dd, IH), 6.99 (dd, IH), 5.15 - 5.09 (m, IH), 4.36 (s, 2H), 4.18 - 4.08 (r

2.29 - 2.22 (m, IH), 2.21 - 2.12 (m, IH), 2.08 - 1.88 (m, 3H), 1.88 - 1.79 (m, IH), 1.7' 1.72 (m, IH), 1.64 - 1.48 (m, 8H).

Example 12: (λ)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(pyra zin-2- 5 ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-But-3-enyl-2-(3-fluoro-phenyl)-hex-5-enoic acid methyl ester

10

(3-Fluoro-phenyl)-acetic acid methyl ester (4.30 g) was dissolved in tetrahydrofuran ('. mL) and cooled to -78°C. Lithium έ>w(trimethylsilyl)amide (25.6 mL, IM THF solutic was added and the solution was stirred for 30 minutes. 4-Bromo-but-l-ene (2.60 mL) ' added and the reaction contents were allowed to warm to room temperature and stir fo i ₅ hour. The reaction mixture was again cooled to -78°C. Lithium /7/5(trimethylsilyl)ami (25.6 mL, IM THF solution) was added and the solution was stirred for 30 minutes. 4- Bromo-1-butene (2.60 mL) was added and the reaction mixture was allowed to warm i room temperature and stir for an hour. The contents were again cooled to -78°C and fi aliquots of Lithium b/.y(trimethylsilyl)amide (25.6 mL, IM THF solution) and 4-brom<

₂ o butene (2.60 mL) were added following the procedure outlined above. After stirring overnight, water (20 mL) was added and the reaction mixture extracted with diethyl et (2 x 60 mL). The combined organic extracts were dried with magnesium sulfate and evaporated. The resulting liquid was purified by column chromatography on silica elui with ethyl acetate / /sσhexane (1 / 99) to afford the sub-titled compound (5.0 g).

2S m/e 277 [M+H] ⁺

To 2-but-3-enyl-2-(3-fluoro-phenyl)-hex-5-enoic acid methyl ester (Example 12a) (5.1

5 in dichloromethane (100 mL) was added Grubbs Catalyst (2nd Generation, Sigma- AIc

Company Ltd) (0.05 g). The mixture was warmed to reflux under nitrogen. After 20 h the reaction was cooled to room temperature, evaporated to an oil and purified by coll chromatography on silica eluting with ethyl acetate / /søhexane (5 / 95) to yield an oil

Analysis of the product showed that significant amounts of starting material was prese io the mixture so the mixture was subjected to a repetition of the reaction conditions and purification as above to afford the subtitled compound as a coloured oil (3.60 g).

m/e 249 [M+H] ⁺

i ₅ c) l-(3-Fluoro-phenyl)-cycloheptanecarboxylic acid methyl ester

l-(3-Fluoro-phenyl)-cyclohept-4-enecarboxylic acid methyl ester (Example 12b) (1.0<o was disolved in methanol (20 mL), palladium on carbon (50 mg) added and mixture st under 4 atm of hydrogen overnight. The solution was filtered and evaporated to afford sub-titled compound (1.09 g).

m/e 251 [M+H] ⁺

d) l-(3-Fluoro-phenyl)-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-y] ester

H l-(3-Fluoro-phenyl)-cycloheptanecarboxylic acid methyl ester (Example 12c) (0.280 was dissolved in toluene (100 mL) and (i?)-quinuclidin-3-ol (0.320 g) was added. Toll

(10 mL) was distilled off in a Dean and Stark apparatus and after cooling sodium hydi

(10 mg) was added. The reaction was refluxed in a Dean and Stark apparatus for 4 hoi after which time an extra amount of sodium hydride (10 mg) was added and the reacti was heated to reflux for a further 4 hours. After allowing to cool to room temperature, toluene was washed with water, dried (MgSO ₄ ) and evaporated. The residue was puri by column chromatography eluting with ethyl acetate / isohexane I triethylamine (50 /

1) then ethyl acetate / triethylamine (99 / 1) to afford the sub-titled compound (0.200 j

m/e 346 [M+H] ⁺ 1H NMR (399.824 MHz, CDCl ₃ ) δ 7.26 (td, IH), 7.10 - 7.07 (m, IH), 7.04 (dd, IH), t (ddd, IH), 4.78 - 4.73 (m, IH), 3.14 (ddd, IH), 2.79 - 2.66 (m, 3H), 2.66 - 2.56 (m, IP 2.53 - 2.46 (m, IH), 2.46 - 2.36 (m, 2H), 2.13 - 1.99 (m, 2H), 1.90 - 1.85 (m, IH), 1.7 1.40 (m, HH), 1.29 - 1.18 (m, IH).

Example 12: (i?)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(pyra zin-2- ylcarbamoylmethyl)-l-azonia-bicydo[2.2.2]octane bromide

l-(3-Fluoro-phenyl)-cycloheptanecarboxylic acid (7?)-(l-aza-bicyclo[2.2.2]oct-3-yl) e (Example 12d) (0.100 g) was dissolved in acetonitrile (8 mL) and 2-bromo-N-pyrazin acetamide (Example 1 Ia) (0.05 g) was added. The reaction mixture was stirred for 3 c diluted with diethyl ether (8 mL), stirred for a futher 10 minutes, the resulting solid w filtered and washed with diethyl ether (3 x 8 mL) to afford a solid which was recrystai from hot butanone (8 mL) to afford the titled compound as a solid (0.081 g).

m/e 481 [M ⁺ ] 1H NMR (399.826 MHz, DMSO-D ₆ ) δ 1 1.42 (s, IH), 9.28 (s, IH), 8.49 - 8.45 (m, 2H 7.40 (td, IH), 7.19 - 7.12 (m, 2H), 7.09 (td, IH), 5.17 - 5.10 (m, IH), 4.40 - 4.30 (m, 2 4.16 - 4.07 (m, IH), 3.71 - 3.57 (m, 4H), 3.52 - 3.41 (m, IH), 2.43 - 2.27 (m, 2H), 2.2l 2.19 (m, IH), 2.19 - 2.09 (m, IH), 2.05 - 1.87 (m, 3H), 1.86 - 1.76 (m, IH), 1.71 - 1.4( 9H).

Example 13: (R)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(isoxa zol-3- ylcarbamoylmethy I)- 1 -azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-./V-isoxazol-3-yl-acetamide

Isoxazol-3-ylamine (1.14 g) was dissolved in dichloromethane (50 mL) and potassium carbonate (3.74 g) was added. Bromoacetyl chloride (1.12 mL) was added slowly wit] stirring and the suspension was stirred overnight. The reaction mixture was washed w water (2 x 50 mL), dried and evaporated. The product was recrystallised from dichloromethane / wohexane to afford the sub-titled compound (2.3 g).

Example 13: (R)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarboπyloxy]-l-(isox azol-3- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octaπe bromide

H

l-(3-Fluoro-phenyl)-cycloheptanecarboxylic acid (7?)-(l-aza-bicyclo[2.2.2]oct-3-yl) e: (Example 12d) (50 mg) and 2-bromo-./V-isoxazol-3-yl-acetamide (Example 13a) (30 re were dissolved in acetonitrile (4 niL) and stirred overnight. The solution was diluted v diethyl ether (12 mL) and stirred overnight. The resulting crystals were filtered off, washed with ether (3 x 10 mL) and dried to afford the titled compound as a solid (48 r

m/e 470 [M ⁺ ]

¹ H NMR (399.826 MHz, DMSO-D ₆ ) δ 1 1.69 (s, IH), 8.90 (d, IH), 7.40 (td, IH), 7.18 7.07 (m, 3H), 6.91 (d, IH), 5.16 - 5.10 (m, IH), 4.31 (d, IH), 4.25 (d, IH), 4.09 (ddd, 3.68 - 3.53 (m, 4H), 3.43 (dd, IH), 2.42 - 2.27 (m, 2H), 2.25 - 2.19 (m, IH), 2.18 - 2.0 IH), 2.04 - 1.88 (m, 3H), 1.85 - 1.75 (m, IH), 1.69 - 1.51 (m, 9H).

Example 14: (/?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride

a) Cycloheptyl-phenyl-methanone

Phenylmagnesium bromide (3.0M solution in diethyl ether) (271 mL), was added dro] to a stirred (overhead stirrer) solution of cycloheptanecarbonitrile (50 g) in 229 mL di ether under nitrogen at such a rate as to maintain gentle reflux. The reaction mixture v then heated at reflux for 3 hours. TLC indicated no starting material present in the rea mixture. The reaction mixture was allowed to cool to room temperature and stood urn nitrogen overnight. The reaction mixture was cooled to 0 ⁰ C and treated dropwise witl mL 4N HCl(aq) keeping the temperature below 2O ⁰ C. 4N sulfuric acid (203 mL) was added dropwise rapidly to start with and then more slowly towards the end. The ice be was removed and the diethyl ether was distilled off. The reaction mixture was heated 80-90 ⁰ C for 3.5 hours then allowed to cool to room temperature and stood overnight, mixture was diluted with ether (approx 450 mL) and water (100 mL). The layers wen separated and the aqueous layer was extracted with ether (2 x 400 mL). The organic 1 were combined and washed with saturated aqueous sodium hydrogen carbonate (600 i and brine (600 mL), dried over magnesium sulphate, filtered and evaporated to give tb sub-titled compound as an orange liquid (86.5 g).

¹ H NMR (300 MHz, CDCl ₃ ) δ 7.96-7.91 (d, 2H), 7.54-7.49 (m, IH), 7.48-7.40 (t, 2H) 3.48-3.37 (m, IH), 1.98-1.88 (m, 2H), 1.85-1.44 (m, 10H). b) ( 1 -Chloro-cyclohepty^-phenyl-methanone

Sulfuryl chloride (210 mL) was added dropwise to neat cycloheptyl-phenyl-methanom (Example 14a) (86.5 g) at 0 ⁰ C over approximately 1 hour. Gas evolution and an exoth were observed. The internal temperature was kept below 15°C during the addition and evolved gas was scrubbed by passing through a 10.2M aqueous solution of NaOH. Tb reaction mixture was heated to reflux overnight. The reaction mixture was cooled to 0' and poured slowly onto ice ( 1 L) with stirring. The layers were separated and the aquc layer was extracted with ether (2 x 400 mL). The combined organic layers were wash with water (600 mL), saturated aqueous sodium hydrogen carbonate (600 mL), and bri

(600 mL), dried over magnesium sulphate, filtered and evaporate to give the sub-titlec compound as a brown oil (100 g).

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.10-8.06 (d, 2H), 7.52-7.46 (t, IH), 7.44-7.36 (t, 2H), (ddd, 2H), 2.29 (ddd, 2H), 1.84-1.73 (m, 2H), 1.68-1.58 (m, 2H), 1.58-1.43 (m, 4H)

c) 1-Phenyl-cycloheptanecarboxylic acid

A solution of (l-chloro-cyclohepty^-phenyl-methanone (Example 14b) (100 g) in 750 dioxane was treated dropwise rapidly with a cloudy solution of silver nitrate (137 g) ir water (85 mL) causing a precipitate to form. The reaction mixture was heated to 75 ⁰ C 4.5 hours. The reaction mixture was cooled to room temperature then filtered and concentrated to approximately 200 mL. Water (200 mL) and ether (300 mL) were adc and the layers separated. The aqueous layer was extracted with ether (2 x 250 mL). T combined organic layers were extracted with 10% aqueous sodium carbonate (3 x 250 mL). The combined basic extracts were heated to 90 ⁰ C over 40 minutes and then cool room temperature and acidified with concentrated HCl (aq). The resulting brown solid filtered off, washed with water (x2) and dried under vacuum at 5O ⁰ C. Crystallisation ft hot ethanol (40 mL) gave the sub-titled compound as pale brown crystals (9.83 g).

¹ H NMR (400 MHz, CD ₃ OD) δ 7.36-7.26 (m, 4H), 7.21-7.15 (m, IH), 2.43-2.35 (m, 2 2.07-1.98 (m, 2H), 1.70-1.53 (m, 8H).

d) 1 -Phenyl -cycloheptanecarboxylic acid methyl ester

A 2.0 M solution of trimethylsilyl diazomethane (29.2 mL) was added dropwise to a solution of 1-phenyl-cycloheptanecarboxylic acid (Example 14c) (9.8 g) in methanol ( mL) and toluene (300 mL) under an atmosphere of nitrogen. After 45 minutes the reaction mixture was concentrated under vacuum and the crude product was purified by column chromotography eluting with 0-10% ethyl acetate / cyclohexane to give the product as a pale yellow oil (9.25 g).

¹ H NMR (300 MHz, CD ₃ OD) δ 7.32-7.24 (m, 4H), 7.21-7.12 (m, IH), 3.60 (s, 3H), 2 2.32 (m, 2H), 2.07-1.96 (m, 2H), 1.65-1.58 (m, 8H).

e) 1 -Phenyl -cycloheptanecarboxylic acid (R)-(I -aza-bicyclo[2.2.2]oct-3-yl) ester

H

A solution of (/?)-(3)-quinuclidinol (10.13 g) and 1-phenyl-cycloheptanecarboxylic aci methyl ester (Example 14d) (9.25 g) in toluene (90 mL) was heated to reflux with a D<

Stark trap for 30 min. The reaction mixture was allowed to cool to room temperature a the trap was removed. Sodium hydride (60% dispersion in mineral oil) (3.19 g) was ad portionwise under nitrogen and the reaction mixture was heated to reflux overnight un nitrogen. The reaction mixture was cooled in an ice bath and diluted with ethyl acetate (200 mL) and water (200 mL). The mixture was filtered and the layers separated. The aqueous layer was extracted with ethyl acetate (2 x 250 mL) and the combined organic layers were washed with brine, dried over magnesium sulfate and evaporated to give tl

crude product which was purified by silica gel chromatography eluting with EtOAc containing 1% triethylamine to give the sub-titled compound as a colourless oil (7.63

¹ H NMR (400 MHz, CD ₃ OD) δ 7.34-7.28 (m, 4H), 7.23-7.17 (m, IH), 4.80-4.75 (m, 3.12 (ddd, IH), 2.75-2.65 (m, 3H), 2.53-2.37 (m, 4H), 2.14-2.06 (m, 2H), 1.88-1.85 IH), 1.69-1.54 (m, 10H), 1.54-1.42 (m, IH), 1.35-1.24 (m, IH).

f) 2-Chloro-./V-pyridin-2-yl-acetamide

A solution of 2-amino-pyridine (1.0 g) in dry dichloromethane (10.6 mL) under nitrog O ⁰ C was treated with triethylamine (1.63 mL) followed by slow addition of chloroacet chloride (0.93 mL). The reaction mixture was allowed to warm up to room temperatur After 2 hours, the mixture was partitioned between dichloromethane and water. The p] were separated and the aqueous layer was extracted with dichloromethane (x2). The combined organic layers were washed with brine, dried over magnesium sulphate, fϊlt< and concentrated to give the crude product which was purified by silica gel chromatography eluting with 0-30% ethyl acetate / cyclohexane to give the title compi (1.43 g) as a pink solid. Further purification was achieved by trituration with 40-60 petroleum ether to give 1.15 g of the desired product. Crystallisation of a 0.94 g portio the material from refluxing acetonitrile (2.4 mL) gave the sub-titled compound as a pii solid (0.73g).

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.96 (s, IH), 8.32 (ddd, IH), 8.21 (d, IH), 7.76 (ddd, IH), 7.12 (ddd, IH), 4.20 (s, 2H).

Example 14: (/?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride

H

A solution of 1-phenyl-cycloheptanecarboxylic acid (R)-( l-aza-bicyclo[2.2.2]oct-3-yl^ ester (Example 14e) (254 mg) in acetonitrile (5 mL) was treated with 2-chloro-N-pyric 2-yl-acetamide (Example 14f) (146 mg) and the resulting yellow solution was stirred a room temperature overnight during which time a solid precipitated. The reaction mixti was treated with ~2 mL of ether and the solid was filtered off, washed with ether and c under vacuum to give the title compound as an off-white solid (217 mg). Crystallisatic from refluxing acetonitrile (20 mL) gave 98 mg of the titled compound as a white crystalline solid, m/e 462 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 11.09 (s, IH), 8.34-8.32 (d, IH), 7.97 (d, IH), 7.! 7.79 (t, IH), 7.33-7.25 (m, 4H), 7.21-7.13 (m, 2H), 5.07 (m, IH), 4.29 (s, 2H), 4.07 (ddd, IH), 3.65-3.51 (m, 4H), 3.41-3.29 (m, IH), 2.36-2.23 (m, 2H), 2.17-2.04 (m, _ 1.99-1.81 (m, 3H), 1.78-1.66 (m, IH), 1.77-1.19 (m, 9H).

Preparation of Example 14: (/?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridh ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride Crystalline Form A

A solution of 1-phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 14e) (254 mg, 0.78mmol) in acetonitrile (5 mL) was treated with 2-chlι N-pyridin-2-yl-acetamide (Example 14f) (146 mg) and the resulting yellow solution w stirred at room temperature overnight during which a solid precipitated. The reaction mixture was treated with a couple of mLs of ether and the solid was filtered off, washe with ether and dried under vacuum to give the title compound (217 mg) as an off- white solid. Crystallisation from refluxing acetonitrile (20 mL) gave 98 mg of the title comp<

m/e 462 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 11.09 (s, IH), 8.34-8.32 (d, IH), 7.97 (d, IH), 7. 7.79 (t, IH), 7.33-7.25 (m, 4H), 7.21-7.13 (m, 2H), 5.07 (m, IH), 4.29 (s, 2H), 4.0^ (ddd, IH), 3.65-3.51 (m, 4H), 3.41-3.29 (m, IH), 2.36-2.23 (m, 2H), 2.17-2.04 (m, 1.99-1.81 (m, 3H), 1.78-1.66 (m, IH), 1.77-1.19 (m, 9H).

Analysis of Example 14: (/?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride Crystalline Form A

A sample of crystalline Example 14 Crystalline Form A obtained by the procedure described above was analysed by XRPD (PANalytical X'Pert system), DSC and TGA.

The melting temperature of Example 14 chloride Form A as determined by DSC was i to be 239°C (onset) (±2°C). Weight loss observed prior to melting by TGA was neglig GVS determination gave a neglible weight increase (%w/w) at 80% RH (±0.2%).

An XRPD spectrum of Example 14 chloride Form A is presented in Figure 1.

Example 15: (R)-3-(l-Phenyl-cycloheptanecarbonyIoxy)-l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-/V-pyridin-2-yl-acetamide

To a solution of 2-aminopyridine (48.8 mmol) in anhydrous THF (98 mL) at room temperature was added Et ₃ N (58.6 mmol) and bromoacetyl bromide (58.6 mmol) drop The mixture was stirred overnight and quenched with sat. NaHCO _{3 (aq} ). EtOAc was ad< to the mixture and the layers separated. The aqueous phase was extracted with EtOAc

Purification by flash silica gel chromatography eluting with 1-2% MeOH / dichloromethane gave the sub-titled compound as a yellow solid (1.14 g).

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.75 (s, IH), 8.26 (ddd, IH), 8.10 (d, IH), 7.67 (ddd, 7.03 (ddd, IH), 3.94 (s, 2H).

Example 15: (/?)-3-(l-Phenyl-cydoheptanecarbonyloxy)-l-(pyridin-2- ylcarbamoylmethyl)-! -azonia-bicyclo[2.2.2]octane bromide

H

1-Phenyl-cycloheptanecarboxylic acid (λ)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Examr. 14e) (0.79 mmol) and 2-bromo-/V-pyridin-2-yl-acetamide (Example 15a) (0.87 mmol) stirred together in anhydrous MeCN at room temperature for 2.5 days. The reaction mixture was concentrated in vacuo and the yellow solid purified by flash silica gel coli chromatography eluting with 2-8% MeOH / dichloromethane to give a tan solid which crystallised from boiling MeCN to give the titled compound as a white solid (211 mg).

m/e 462 [M] ⁺ 1H NMR (400 MHz, DMSO-D ₆ ) δ 1 1.02 (s, IH), 8.33 (ddd, IH), 7.97 (d, IH), 7.86-' (m, IH), 7.32-7.25 (m, 4H), 7.23-7.12 (m, 2H), 5.09-5.04 (m, IH), 4.23 (s, 2H), 4.0 (ddd, IH), 3.63-3.49 (m, 4H), 3.41-3.29 (m, IH), 2.37-2.22 (m, 2H), 2.17-2.04 (m, : 1.98-1.83 (m, 3H), 1.78-1.66 (m, IH), 1.65-1.39 (m, 9H).

Preparation of Example 15: (λ)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridii ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide Crystalline Form A

1-Phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Exam] 14e) (0.79 mmol) and 2-bromo-./V-pyridin-2-yl-acetamide (Example 15a) (0.87 mmol) stirred together in anhydrous MeCN at room temperature for 2.5 days. The reaction mixture was concentrated in vacuo and the yellow solid purified by flash silica gel col 5 chromatography eluting with 2-8% MeOH/dichloromethane to give a tan solid. The sc was dissolved up in refluxing MeCN and the solution was allowed to cool down to roc temperature. The resulting crystals were filtered off and washed with a small quantity cold MeCN to give the title compound (211 mg) as a white crystalline solid. o m/e 462 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 11.02 (s, IH), 8.33 (ddd, IH), 7.97 (d, IH), 7.86- (m, IH), 7.32-7.25 (m, 4H), 7.23-7.12 (m, 2H), 5.09-5.04 (m, IH), 4.23 (s, 2H), 4.C (ddd, IH), 3.63-3.49 (m, 4H), 3.41-3.29 (m, IH), 2.37-2.22 (m, 2H), 2.17-2.04 (m, _ 1.98-1.83 (m, 3H), 1.78-1.66 (m, IH), 1.65-1.39 (m, 9H). 5

Analysis of Example 15: (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide Crystalline Form A

A sample of crystalline Example 15 Crystalline Form A obtained by the procedureo described above was analysed by XRPD (PANalytical X'Pert system), DSC and TGA.

The melting temperature of Example 15 bromide Form A as determined by DSC was found to be 230 ⁰ C (onset) (±2°C). Weight loss observed prior to melting by TGA was negligible. GVS determination gave a neglible weight increase (%w/w) at 80% RH5 (±0.2%).

An XRPD spectrum of Example 15 bromide Form A is presented in Figure 2.

0 Example 16: (R)-3-(l-PhenyI-cydoheptanecarbonyIoxy)-l-(pyridin-4-

a) 2-Chloro-./V-pyridin-4-yl-acetamide

A suspension of 4-aminopyridine (0.96 g) in dry dichloromethane (10 mL) under nitre was cooled to 0 ⁰ C in an ice bath. Triethylamine (1.56 mL) was added, followed by th slow addition of chloroacetyl chloride (0.89 mL). The ice bath was removed and the reaction mixture allowed to reach room temperature. The reaction mixture was diluted water (20 mL) and dichloromethane (25 mL). The solid was filtered off, washed with pentane and dried to give the title compound as a brown solid (0.87 g). The layers of tl filtrate were separated and the organic layer was washed with water, dried and the soh was evaporated to give a dark brown glass. Trituration with pentane gave another bate the title compound (0.91 g).

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 10.79 (s, IH), 8.47 (d, 2H), 7.59 (d, 2H), 4.33 (s, 2

Example 16: (/f)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-4- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride

H

2-Chloro-yV-pyridin-4-yl-acetamide (Example 16a) (30 mg) was added to a solution of phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example (53 mg) in acetonitrile (1 mL). The reaction mixture was stirred at room temperature f< h. Diethyl ether (2 mL) was added and the reaction mixture was filtered to give a light brown solid. The solid was washed several times with diethyl ether and dried under vacuum at 40 ⁰ C. Purification by column chromatography eluting with 0-10% MeOH /

m/e 462 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 11.34 (s, IH), 8.46 (d, 2H), 7.55 (d, 2H), 7.34-7.: (m, 4H), 7.22-7.17 (m, IH), 5.08 (m, IH), 4.30 (s, 2H), 4.11-4.02 (m, IH), 3.65-3. (m, 4H), 3.42-3.30 (m, IH), 2.38-2.24 (m, 2H), 2.17-2.06 (m, 2H), 1.99-1.84 (m, 3] 1.79-1.67 (m, IH), 1.69-1.26 (m, 9H).

Example 17: (/f)-l-[(5-FIuoro-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phenyl - cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-N-(5-fluoro-pyridin-2-yl)-acetamide

The title compound (0.99 g, 73%, white solid) was prepared according to the method i in Example 14f but using 2-amino-5-fluoro-pyridine.

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 10.91 (s, IH), 8.35 (d, IH), 8.10 (dd, IH), 7.80-7. (m, IH), 4.34 (s, 2H).

Example 17: (/?)-l-[(5-Fluoro-pyridin-2-yIcarbamoyl)-methyl]-3-(l-phenyl - cycloheptanecarbonyloxy)- 1 -azonia-bicyclo[2.2.2]octane chloride

H

2-Chloro-./V-(5-fluoro-pyridin-2-yl)-acetamide (Example 17a) (31 mg) was added to a solution of l -nhenvl-cvclnhe.ntane.rarhr>γvlir arid ( R}-( 1 9 e>c

temperature overnight. Diethyl ether (2 mL) was added to the reaction mixture and th( white solid was filtered off, washed several times with diethyl ether and dried under vacuum at 4O ⁰ C to give the title compound (49 mg).

m/e 480 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 11.19 (s, IH), 8.36 (d, IH), 8.02 (m, IH), 7.81 (d IH), 7.33-7.26 (m, 4H), 7.22-7.17 (m, IH), 5.07 (m, IH), 4.26 (s, 2H), 4.11-4.03 (i IH), 3.64-3.50 (m, 4H), 3.41-3.29 (m, IH), 2.36-2.23 (m, 2H), 2.17-2.05 (m, 2H), 1.82 (m, 3H), 1.78-1.65 (m, IH), 1.70-1.41 (m, 9H).

Example 18: (/?)-l-[(5-Methyl-pyridin-2-ylcarbamoyl)-methyI]-3-(l-phenyl - cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-/v ^' -(5-methyl-pyridin-2-yl)-acetamide

The title compound (0.50 g) was prepared according to the method used in Example 1 but using 2-amino-5-picoline.

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 10.69 (s, IH), 8.17 (dt, IH), 7.95 (d, IH), 7.63 (dc IH), 4.32 (s, 2H), 2.25 (s, 3H).

Example 18: (/?)-l-[(5-MethyI-pyridin-2-ylcarbamoyl)-methyI]-3-(l-phenyl - cycloheptanecarbonyloxy)- 1 -azonia-bicy clo[2.2.2]octane chloride

H

The title compound (36 mg) was prepared according to the method used to prepare Example 17 using 2-chloro-/V-(5-methyl-pyridin-2-yl)-acetamide (Example 18a) in pi; of 2-chloro-./V-(5-fluoro-pyridin-2-yl)-acetamide.

m/e 476 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 10.98 (s, IH), 8.17 (d, IH), 7.88 (d, IH), 7.65 (dc IH), 7.33-7.25 (m, 4H), 7.23-7.17 (m, IH), 5.07 (m, IH), 4.24 (s, 2H), 4.10-4.02 (i IH), 3.64-3.50 (m, 4H), 3.40-3.27 (m, IH), 2.37-2.22 (m, 2H), 2.23 (s, 3H), 2.17-2 (m, 2H), 1.97-1.84 (m, 3H), 1.78-1.66 (m, IH), 1.66-1.35 (m, 9H).

Example 19: (/?)-3-(l-Phenyl-cycIoheptanecarbonyloxy)-l-(pyridin-3- y lcarbamoy lmethyl)- 1 -azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-./V-pyridin-3-yl-acetamide

A mixture of 3-aminopyridine (350 mg) and sodium hydroxide (0.6 g) were dissolved water (8 mL) and the reaction mixture was cooled in an ice bath. Chloroacetyl chloric (1.19 mL) was added dropwise and the reaction mixture was allowed to stir at room temperature overnight. The reaction mixture was extracted with dichloromethane and organic layer was concentrated and purified by column chromatography, eluting with ' 60% ethyl acetate / cyclohexane to give the title compound (0.1Og) as a white solid.

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 10.51 (s, IH), 8.73 (d, IH), 8.30 (dd, IH), 8.03 (d< IH), 7.40-7.35 (m, IH), 4.30 (s, 2H).

Example 19: (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-3- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride

H

The title compound (78 mg) was prepared by an analogous method to that used in Exe 15 using 2-chloro-/V-pyridin-3-yl-acetamide in place of 2-bromo-./V-pyridin-2-yl-aceta

m/e 462 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ) δ 11.27 (s, IH), 8.76 (d, IH), 8.30 (dd, IH), 7.98 (c IH), 7.37 (ddd, IH), 7.33-7.25 (m, 4H), 7.22-7.15 (m, IH), 5.07 (d, IH), 4.28 (dd, 4.1 1-4.03 (m, IH), 3.65-3.50 (m, 4H), 3.41-3.29 (m, IH), 2.37-2.21 (m, 2H), 2.19-2 (m, 2H), 1.97-1.83 (m, 3H), 1.78-1.66 (m, IH), 1.71-1.27 (m, 9H).

Example 20: (/?)-l-[(2-Methyl-pyridin-4-ylcarbamoyl)-methyl]-3-(l-phenyl - cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-iV-(2-methyl-pyridin-4-yl)-acetamide

The title compound (1.0 g) was prepared according to the method used in Example 14 using 4-amino-2-methylpyridine.

¹ H NMR C400 MH/. DMSO-Dλϊ δ 10.64 fs. I HV 8.32 Cd. IHV 7.44 Cd. IHV 7.38-7.3

Example 20: (R)-l-[(2-Methyl-pyridin-4-ylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

H

The title compound was prepared using an analogous procedure to that used to prepaπ Example 17. Further purification was achieved by silica gel chromatography eluting w 0-20% MeOH / dichloromethane to give the title compound as a white solid (57 mg).

m/e 476 [M] ⁺ 0 ¹ H NMR (400 MHz, DMSO-D ₆ ) δ 11.32 (s, IH), 8.31 (d, IH), 7.43 (d, IH), 7.35-7.: (m, 5H), 7.22-7.16 (m, IH), 5.09-5.04 (m, IH), 4.30 (dd, 2H), 4.09-4.01 (m, IH), 3 3.49 (m, 4H), 3.41-3.29 (m, IH), 2.38 (s, 3H), 2.39-2.23 (m, 2H), 2.17-2.05 (m, 2H 1.97-1.82 (m, 3H), 1.78-1.65 (m, IH), 1.65-1.41 (m, 9H). 5 Example 21: (R)-l-Phenylcarbamoylmethyl-3-(l-phenyl-cycloheptanecarbonyl ox azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-/V-phenyl-acetamide

o To a solution of bromoacetyl bromide (9.6 mL) and potassium carbonate ( 1 1.4 g) in dichloromethane (100 mL) was added aniline (5 mL) dropwise over 15-20 mins causii the reaction mixture to get warm and a white precipitate to form. After 4.5h the reactic mixture was poured into water, shaken for several minutes and then the phases were ςpnaratpH The nrαanir laver was washed with water and concentrated rn a smaller vnl

resulting in precipitation of a solid that was filtered off to give the sub-titled compouπ (970 mg) as a white solid.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.10 (s, IH), 7.53 (d, 2H), 7.40-7.33 (m, 2H), 7.17 (t. 5 4.03 (s, 2H).

Example 21: (R)-l-PhenyIcarbamoylmethyl-3-(l-phenyl-cycloheptanecarbonyl ox azonia-bicyclo[2.2.2]octane bromide

H io To 1-phenyl-cycloheptanecarboxylic acid (,/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester

(Example 14e) (50 mg) in acetonitrile (1 mL) was added 2-bromo-N-phenylacetamide (Example 21a) (36 mg). The reaction mixture was stirred at room temperature for 3 di Ether was added to the reaction mixture and the resultant solid was collected by filtrat and dried to afford the title compound as a colourless solid (39 mg).

I ₅ m/e 461 [M] ⁺

¹ H NMR (DMSO-D ₆ ): δ 10.49 (s, IH), 7.53-7.50 (m, 2H), 7.35-7.24 (m, 6H), 7.21-7.1 (m, IH), 7.12-7.07 (m, IH), 5.08 (m, IH), 4.21-4.1 1 (m, 2H), 4.06 (dd, IH), 3.64-3.4' 4H), 3.27 (s, IH), 2.37-2.21 (m, 2H), 2.18-2.04 (m, 2H), 1.98-1.88 (m, 3H), 1.77-1.66 20 IH), 1.70-1.30 (m, 9H).

Example 22: (R)-3-(l-Phenyl-cycIoheptanecarbonyloxy)-l-(pyrimidin-4- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride

25 a) 2-Chloro-./V-pyrimidin-4-yl-acetamide

A solution of chloroacetyl chloride (1.22 mmol) in anhydrous CHCl ₃ (2.4 mL) was ad slowly to a mixture of 4-aminopyrimidine (1.1 1 mmol) and Et ₃ N (1.66 mmol) in anhydrous CHCl ₃ (22 mL) at room temperature. The bright yellow mixture gradually turned orange and after 4 hr the reaction was quenched with H ₂ O (1 mL). After stirrir 15 min the mixture was concentrated to dryness under reduced pressure and the residi purified by flash silica gel chromatography (1-2% MeOH / dichloromethane) to give i yellow solid (122 mg).

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 1 1.21 (s, IH), 8.93-8.90 (m, IH), 8.70 (d, IH), 8. (dd, IH), 4.40 (s, 2H).

Example 22: (λ)-3-(l-PhenyI-cycIoheptanecarbonyloxy)-l-(pyrimidin-4- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride

H

1-Phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Exam] 14e) (0.44 mmol) and 2-chloiO-/V-pyrimidin-4-yl-acetamide (Example 22a) (0.48 mm anhydrous MeCN (2 mL) were stirred together at room temperature for 2.5 days. The reaction mixture was concentrated in vacuo and the residue purified by flash silica gel chromatography (2-10% MeOH/dichloro methane) to give the title compound as a ligh yellow solid (134 mg).

m/e 463 [M] ⁺

IXJ XTλ/TD fλ(λC\ \λU-* Flλ/ICπ fVV X 1 1 λλ (<- 1 U\ 9 O l ^ 1 Uλ Q T") (A 1 \λ\ ^ Qλ (A

(m, IH), 3.61-3.49 (m, 4H), 3.40-3.28 (m, IH), 2.36-2.21 (m, 2H), 2.18-2.04 (m, 21 2.00-1.84 (m, 3H), 1.75-1.66 (m, IH), 1.66-1.39 (m, 9H).

Example 23: (R)-l-[(2-Fluoro-phenylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-./V-(2-fluoro-phenyl)-acetamide

To a mixture of 2-fluoroaniline (2 mL) and potassium carbonate (4.3 g) in dichlorome (50 mL) was added bromoacetyl bromide (3.6 mL). The reaction mixture was stirred f then water was added and the phases separated. The organic layer was concentrated, tl residue treated with ether and evaporated again to give the sub-titled compound (4.98 a cream solid that was used without further purification. 1H NMR (400 MHz, CDCl ₃ ): δ 8.38 (s, IH), 8.26 (t, IH), 7.18-7.09 (m, 3H), 4.05 (s,

Example 23: (/?)-l-[(2-Fluoro-phenylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

H

A mixture of 1-phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 14e) (56 mg) and 2-bromo-./V-(2-fluoro-phenyl)-acetamide (Example 2 (44 mg) in acetonitrile (1 mL) was stirred at room temperature for 30 h. The resulting precipitate was collected by filtration, washed with ether and dried under vacuum at 5ι to give the title compound (52 mg) as a colourless solid.

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 10.34 (s, IH), 7.83-7.77 (m, IH), 7.32-7.16 (m, I 5.12-5.03 (m, IH), 4.25 (s, 2H), 4.10-4.02 (m, IH), 3.63-3.51 (m, 4H), 3.41-3.29 (n IH), 2.37-2.23 (m, 2H), 2.17-2.06 (m, 2H), 1.98-1.88 (m, 3H), 1.79-1.67 (m, IH), 1.39 (s, 9H).

Example 24: (R)-l-[(2,3-Difluoro-phenyIcarbamoyI)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-/V-(2,3-difluoro-phenyl)-acetamide

To a mixture of 2,3-difluoroaniline (630 mg) and potassium carbonate (1.01 g) in dichloromethane (30 mL) was added bromoacetyl bromide (0.86 mL). The reaction mixture was stirred for 5h then water was added and the phases separated. The organic layer was concentrated to give a 2: 1 mixture of the sub-titled compound and bromoaα bromide. The residue was dissolved up in dichloromethane and washed with water. Tl volatiles were evaporated to give the sub-titled compound (1.15 g) as an off-white sol that was used without further purification.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.38 (s, IH), 8.03 (t, IH), 7.12-7.05 (m, IH), 7.00-6. ¹ (m, IH), 4.05 (d, 2H).

Example 24: (/f)-l-[(2,3-Difluoro-phenylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

The title compound (colourless solid, 28 mg, 31%) was prepared by a similar procedu that used for Example 23 using 2-bromo-./V-(2,3-difluoro-phenyl)-acetamide (Example in place of 2-bromo-/V-(2-fluoro-phenyl)-acetamide.

m/e 497 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 10.54 (s, IH), 7.58 (t, IH), 7.32-7.16 (m, 7H), 5. 5.04 (m, IH), 4.26 (s, 2H), 4.10-4.02 (m, IH), 3.62-3.49 (m, 4H), 3.42-3.29 (m, IH 2.37-2.23 (m, 2H), 2.17-2.06 (m, 2H), 1.98-1.85 (m, 3H), 1.79-1.67 (m, IH), 1.66-1 (m, 9H).

Example 25: (/?)-l-[2-(2,3-Dihydro-benzofuran-5-yl)-ethyl]-3-(l-phenyI- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane formate

H A mixture of 1-phenyl-cycloheptanecarboxylic acid (λ)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 14e) (126 mg) and 5-(2-bromo-ethyl)-2,3-dihydro-benzofuran (105 m| 0.46mmol) in acetonitrile (1.5 mL) was stirred at room temperature for 22h. The volat were evaporated and the residue purified by silica gel chromatography eluting with dichloromethane then 5% then 10% MeOH/dichloromethane. The relevant fractions w combined and evaporated and the residue triturated with dichloromethane to give an o white foam. Further purification was achieved by reverse-phase HPLC (5-98% MeCN containing 0.1% formic acid) to give the title compound (70 mg) as a white gummy so

m/e 474 [M] ⁺ 1H NMR (400 MHz, DMSO-D ₆ ): δ 8.35 (s, IH), 7.35-7.26 (m, 4H), 7.24-7.18 (m, IF

IH), 3.47-3.25 (m, 5H), 3.20-3.06 (m, 3H), 3.04-2.97 (m, IH), 2.86-2.75 (m, 2H), 2.23 (m, 2H), 2.18-2.10 (m, 2H), 2.01-1.78 (m, 3H), 1.69-1.44 (m, 10H).

Example 26: (R)-l-[2-(4-Fluoro-phenoxy)-ethyl]-3-(l-phenyI- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane formate

H

A mixture of 1 -phenyl-cycloheptanecarboxylic acid (7?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 14e) (50 mg) and l-(2-bromoethoxy)-4-fluorobenzene (50 mg) in acetonitrile (1 mL) was stirred at room temperature for 22h. Purification by prep. HPL using 5-98% MeCN/H ₂ O containing 0.1% formic acid gave the title compound (19 mj a colourless oil.

m/e 466 [M] ⁺ 1H NMR (400 MHz, DMSO-D ₆ ): δ 8.39 (s, IH), 7.28-7.23 (m, 4H), 7.20-7.11 (m, 3P 6.97-6.92 (m, 2H), 5.06-4.99 (m, IH), 4.39-4.28 (m, 2H), 3.93 (ddd, IH), 3.70-3.56 2H), 3.56-3.46 (m, 4H), 3.15-3.03 (m, IH), 2.35-2.20 (m, 2H), 2.15-2.03 (m, 2H), 1.78 (m, 3H), 1.73-1.61 (m, IH), 1.61-1.39 (m, 9H).

Example 27: (R)-3-(l-phenyl-cycloheptanecarbonyIoxy)-l-(pyridazin-4- ylcarbamoylmethy I)- 1 -azonia-bicyclo[2.2.2]octane formate

a) 2-Chloro-yV-pyridazin-4-yl-acetamide

A solution of pyridazin-4-ylamine (1.0 g) in dry dichloromethane (10 niL) under nitre was cooled to 0 ⁰ C in an ice bath. Triethylamine (1.6 mL) was added, followed by slo addition of chloroacetyl chloride (0.92 mL). On completion of the addition the ice bat removed and the reaction mixture was allowed to reach room temperature and stirred hours. The reaction mixture was diluted with water (25 mL) and dichloromethane (30 A solid was filtered off and washed with pentane, water and more pentane to give the titled compound (0.87g, 48%) as a brown solid.

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 1 1.17 (s, IH), 9.33 (dd, IH), 9.07 (dd, IH), 7.94 IH), 4.39 (s, 2H).

Example 27: (R)-3-(l-phenyl-cycloheptanecarbonyloxy)-l-(pyridaziπ-4- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane formate

H

2-Chloro-iV-pyridazin-4-yl-acetamide (Example 27a) (58 mg) was added to a solution phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example (100 mg) in acetonitrile (2 mL). The reaction mixture was allowed to stir at room temperature for 24h. Diethyl ether (2 mL) was added to the reaction mixture and stirre 15 minutes. The solid was filtered off, washed with diethyl ether, and dried under vac at 40 ⁰ C overnight. The filtrate was evaporated, combined with the solid and purified b column chromatography, eluting with 0-15% MeOH/dichloromethane. Further purific by reverse-phase prep HPLC with gradient elution from 15% MeCN/H ₂ O containing formic acid increasing by 1% per minute gave the title compound (19 mg) as a colour gum.

¹ H NMR (400 MHz, DMSO-D ₆ ): 6 9.17 (s, IH), 8.89 (d, IH), 8.41 (s, IH), 7.80 (dd IH), 7.32-7.25 (m, 4H), 7.21-7.15 (m, IH), 5.08-5.02 (m, IH), 4.28-4.14 (m, 2H), (dd, IH), 3.66 (d, 2H), 3.62-3.48 (m, 2H), 3.45-3.33 (m, IH), 2.37-2.23 (m, 2H), 2 2.05 (m, 2H), 1.96-1.83 (m, 3H), 1.75-1.62 (m, IH), 1.56-1.42 (m, 9H).

Example 28: (/?)-l-[(5-Fluoro-pyridin-3-ylcarbamoyl)-methyl]-3-(l-phenyl - cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-/V-(5-fluoro-pyridin-3-yl)-acetamide

A solution of 3-amino-5-fluoropyridine (1 g) in dry dichloromethane (10 mL) under nitrogen was cooled to 0 ⁰ C in an ice bath. Triethylamine (1.36 mL) was added follow by slow addition of chloroacetyl chloride (0.78 mL). On completion of the addition, tl bath was removed and the reaction mixture was allowed to reach room temperature an was stirred for 2 hours. The reaction mixture was diluted with water (25 mL) and dichloromethane (30 mL). The organic layer was washed with water (2 x 20 mL), drii (MgSO ₄ ) and evaporated to give the crude product. Purification was achieved by silic; chromatography eluting with 0-30% ethyl acetate/cyclohexane to give the sub-titled compound (1.0 g) as a tan coloured solid.

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 10.77 (s, IH), 8.56 (t, IH), 8.33 (d, IH), 8.04 (dt. 4.33 (s, 2H).

Example 28: (R)-l-[(5-Fluoro-pyridin-3-ylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

H

2-Chloro-N-(5-fluoro-pyridin-3-yl)-acetamide (Example 28a) (53 mg) was added to a solution of 1-phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) e (Example 14e) (84 mg) in acetonitrile (2 mL). The reaction mixture was allowed to si 5 room temperature for 24h. The solid was filtered off and dried under vacuum at 40 ⁰ C give the title compound (47 mg, 35%) as a white solid.

m/e 480 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 11.53 (s, IH), 8.58 (s, IH), 8.34 (d, IH), 7.99 (d io IH), 7.32-7.25 (m, 4H), 7.21-7.15 (m, IH), 5.09-5.04 (m, IH), 4.29 (s, 2H), 4.10-4 (m, IH), 3.63-3.51 (m, 4H), 3.41-3.29 (m, IH), 2.37-2.23 (m, 2H), 2.16-2.05 (m, 21 1.96-1.83 (m, 3H), 1.78-1.65 (m, IH), 1.65-1.39 (m, 9H).

Example 29: (/?)-3-(l-phenyl-cycloheptanecarbonyloxy)-l-[2-(pyridin-3-yl oxy)-et i ₅ l-azonia-bicyclo[2.2.2]octane formate

a) 2-(Pyridin-3-yloxy)-ethanol

The sub-titled compound (0.99g, 63%) was prepared according to the procedure descr 20 in WO2004/000829.

b) 3-(2-Bromo-ethoxy)-pyridine

A solution of 2-(pyridin-3-yloxy)-ethanol (200 mg) in 5 mL dichloromethane was coc to O ⁰ C and treated with carbon tetrabromide (524 mg) followed by triphenylphosphim mg) portionwise. The reaction mixture was stirred at O ⁰ C for 30 mins then at room temperature for 45 mins. The volatiles were evaporated and the residue purified by sil gel chromatography eluting with 0-100% EtO Ac/pen tane to give the title compound ( mg) as a colourless liquid which was used without further purification.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.34 (dd, IH), 8.28-8.22 (m, IH), 7.25-7.20 (m, 2H), (t, 2H), 3.66 (t, 2H).

Example 29: (/?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-[2-(pyridin-3-yl oxy)-el l-azonia-bicyclo[2.2.2]octane formate

H

The title compound (10 mg, 13%, colourless gum) was prepared using a procedure analogous to that used for the preparation of Example 25 using 3-(2-Bromo-ethoxy)- pyridine (Example 29b) in place of 5-(2-bromo-ethyl)-2,3-dihydro-benzofuran.

m/e 449 [M] ⁺

¹ H NMR (400 MHz, DMSO-D6): 6 8.51 (s, IH), 8.29 (d, IH), 8.21 (dd, IH), 7.38-7 (m, 2H), 7.28-7.24 (m, 4H), 7.19-7.14 (m, IH), 5.06-4.99 (m, IH), 4.51-4.39 (m, 21

3.99-3.90 (m, IH), 3.74-3.59 (m, 2H), 3.57-3.37 (m, 3H), 3.14-3.05 (m, IH), 2.36-2

(m, 2H), 2.14-2.05 (m, 2H), 1.98-1.82 (m, 3H), 1.74-1.62 (m, IH), 1.62-1.38 (m, 91

Example 30: (R)-l-[(6-Methyl-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

The sub-titled compound (0.95g, 58%, white solid) was prepared by a similar procedu that used for the preparation of Example 28a using 2-amino-6-picoline in place of 3- amino-5-fluoropyridine.

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 10.73 (s, IH), 7.86 (d, IH), 7.69 (t, IH), 7.03-6.9 IH), 4.32 (s, 2H), 2.41 (s, 3H).

Example 30: (/?)-l-[(6-Methyl-pyridin-2-yIcarbamoyl)-methyl]-3-(l-phenyl - cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

H

The title compound (34 mg, 43%, white solid) was prepared using a similar method to used to prepare Example 17 using 2-chloro-N-(6-rnethyl-pyridin-2-yl)-acetamide (Exa 30a) in place of 2-chloro-./V-(5-fluoro-pyridin-2-yl)-acetamide.

m/e 476 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 11.01 (s, IH), 7.79 (d, IH), 7.71 (t, IH), 7.32-7.: (m, 4H), 7.23-7.16 (m, IH), 7.02 (d, IH), 5.09-5.04 (m, IH), 4.24 (s, 2H), 4.06 (dd IH), 3.63-3.50 (m, 4H), 3.38-3.29 (m, IH), 2.38 (s, 3H), 2.38-2.23 (m, 2H), 2.17-2 (m, 2H), 2.00-1.82 (m, 3H), 1.78-1.66 (m, IH), 1.65-1.40 (m, 9H).

Example 31: (jR)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(o-tolylcarbamoy l-met l-azonia-bicyclo[2.2.2]octane bromide

The sub-titled compound (0.83g, 49%, off-white solid) was prepared using a procedui similar to that used to prepare Example 28a using ø-toluidine in place of 3-amino-5- fluoropyridine.

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 9.62 (s, IH), 7.39 (d, IH), 7.23-7.08 (m, 3H), 4.2 2H), 2.23 (s, 3H).

Example 31: (R)-3-(l-Phenyl-cycloheptanecarbonyIoxy)-l-(ø-tolyIcarbamoy l-met l-azonia-bicyclo[2.2.2]octane bromide

H

The title compound (25 mg, 35%, white solid) was prepared using a similar method to used to prepare Example 17 using 2-chloro-/V-ø-tolyl-acetamide (Example 31a) in pla< 2-chloro-/V-(5-fluoro-pyridin-2-yl)-acetamide.

m/e 475 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 10.26 (s, IH), 7.34 (dd, IH), 7.31-7.25 (m, 4H), 7.09 (m, 4H), 5.11-5.05 (m, IH), 4.31 (dd, 2H), 4.07 (ddd, IH), 3.68-3.52 (m, 4H), 3.40-3.31 (m, IH), 2.38-2.22 (m, 2H), 2.19 (s, 3H), 2.17-2.06 (m, 2H), 1.97-1.84 (rr 3H), 1.79-1.67 (m, IH) 1.65-1.39 (m, 9H).

Example 32: (R)-3-(l-Phenyl-cycloheptanecarboπyloxy)-l-(2-pyrazin-2-yl- ethyI)- azonia-bicyclo[2.2.2]octane bromide

To a solution of 2-(2'-hydroxyethyl)pyrazine (0.91 g) in 30 niL dichloromethane at 0°( was added carbon tetrabromide (2.65 g) followed by triphenylphosphine (2.1 g) portionwise. The solution became very dark. After stirring for Ih the reaction mixture adsorbed onto HMN diatomaceous earth and purified by silica gel chromatography eli with 0-5% MeOH/dichloromethane to give a cream solid (3.07g) that was purified furl by silica gel chromatography eluting with 0-50% EtOAc/pentane to give the sub-titled compound (0.47g, 34%) as a yellow oil.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.56-8.48 (m, 3H), 3.78 (t, 2H), 3.37 (t, 2H).

Example 32 : (R)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(2-pyrazin-2-yl-ethyl)- azonia-bicyclo[2.2.2]octane bromide

H 2-(2-Bromo-ethyl)-pyrazine (Example 32a) (43 mg) was added to a solution of 1-phei cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 14e) (5 mg) in acetonitrile (1 mL). The reaction mixture was allowed to stir at room temperat for 68h. The volatiles were evaporated and the product was purified by silica gel chromatography eluting with 0-20% MeOH/dichloromethane. The relevant fractions v combined and concentrated, dissolved up in dichloromethane, filtered and evaporated give the title compound (30 mg) as a yellow gummy glass.

m/e 434 [M] ⁺

¹ H NMR (400 MHz, CD ₃ OD): δ 8.60 (d, IH), 8.54 (dd, IH), 8.51 (d, IH), 7.31-7.24

3H), 3.24-3.12 (m, 3H), 3.08-2.97 (m, IH), 2.36-2.19 (m, 2H), 2.15-2.06 (m, 2H), 1.78 (m, 3H), 1.71-1.59 (m, IH), 1.57-1.37 (m, 9H).

Example 33: (S)-l-(3-Phenoxy-propyl)-3-(l-phenyl-cycloheptanecarbonyloxy )-l- 5 azonia-bicyclo[2.2.2]octane formate

a) 1 -Phenyl -cycloheptanecarboxylic acid (S)-(I -aza-bicyclo[2.2.2]oct-3-yl) ester

A solution of (S)-(+)-3-quinuclidinol (299 mg) and 1-phenyl-cycloheptanecarboxylic ;o methyl ester (Example 14d) (455 mg) in 6.5 mL dry toluene under nitrogen was treate with a 60% dispersion of sodium hydride (94 mg) and the mixture was heated to reflu;

24h then cooled to room temperature and allowed to stand over the weekend. EtOAc a sat. NaHCO _{3 (aq} ) were added and the phases separated. The aqueous phase was extracti with EtOAc (x3) and the combined organic layer was washed with brine, dried (MgSCs and evaporated to afford the crude product. Purification was achieved by silica gel chromatography eluting with 0-10% MeOH/dichloromethane to give the sub-titled compound (384 mg) as a yellow oil.

¹ H NMR (300 MHz, CD ₃ OD): δ 7.39-7.27 (m, 4H), 7.25-7.14 (m, IH), 4.80 (dt, IH).o 3.16 (ddd, IH), 2.83-2.64 (m, 3H), 2.56-2.34 (m, 4H), 2.15-2.04 (m, 2H), 1.91-1.86 IH), 1.72-1.44 (m, HH), 1.38-1.25 (m, IH).

Example 33: (S)-l-(3-Phenoxy-propyl)-3-(l-phenyl-cycloheptanecarbonyloxy )-l- azonia-bicyclo[2.2.2]octane formate

A mixture of 3-phenoxypropyl bromide (0.026 mL) was added to a solution of 1-phen cycloheptanecarboxylic acid (S)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 33a) (' mg) in acetonitrile (1 mL). The reaction mixture was allowed to stir at room temperat for 72hrs then heated to 5O ⁰ C for 3 days. The volatiles were evaporated and the residui purified by silica gel chromatography eluting with 0-10% MeOH/dichloro methane to j 57 mg of a hygroscopic foam. Further purification was achieved by reverse phase HPI using a C18 column eluting with 5-95% MeCNZH ₂ O containing 0.1% formic acid ovei 30mins gave the title compound (43 mg) as an oil.

m/e 462 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 8.38 (s, IH), 7.34-7.24 (m, 6H), 7.23-7.17 (m, H- 6.94-6.87 (m, 3H), 5.07-4.97 (m, IH), 3.98 (t, 2H), 3.87-3.77 (m, IH), 3.44-3.26 (m 5H), 3.16-3.09 (m, IH), 3.01-2.90 (m, IH), 2.38-2.23 (m, 2H), 2.16-1.77 (m, 7H), 1.43 (m, 10H).

Example 34: (R)-l-{[2-(3-Fluoro-phenoxy)-ethylcarbamoyl]-methyl}-3-(l-ph enyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-W-[2-(3-fluoro-phenoxy)-ethyl]-acetamide

A mixture of 2-(3-fluorophenoxy)ethylamine (0.93 g) and potassium carbonate (1.24 ; dichloromethane (20 mL) at O ⁰ C was treated with bromoacetyl bromide (1.04 mL). Tt reaction mixture was stirred at O ⁰ C for 30mins then at room temperature for 6h. Watei added and the mixture was stirred until effervescence ceased. The organic layer was separated and the organic layer was evaporated to afford the crude product which was purified by silica gel chromatography eluting with 0-50% EtOAc/pentane to give the 5 titled compound (1.17 g) as a brown oil.

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.24 (dt, IH), 6.90 (s, IH), 6.71-6.66 (m, 2H), 6.63 (< IH), 4.06 (t, 2H), 3.90 (s, 2H), 3.71 (q, 2H).

Example 34: (/?)-l-{[2-(3-Fluoro-phenoxy)-ethylcarbamoyl]-methyl}-3-(l-p henyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

H The title compound (70 mg, 76%, yellow foam) was prepared by a similar procedure t that used to prepare Example 32 using 2-bromo-./V-[2-(3-fluoro-phenoxy)-ethyl]-acetai (Example 34a) in place of 2-(2-bromo-ethyl)-pyrazine.

m/e 523 [M] ⁺ 1H NMR (400 MHz, DMSO-D ₆ ): δ 8.74 (t, IH), 7.33-7.23 (m, 5H), 7.23-7.16 (m, Ih 6.79-6.71 (m, 3H), 5.09-5.02 (m, IH), 4.03-3.94 (m, 5H), 3.57-3.40 (m, 6H), 3.33-3 (m, IH), 2.35-2.20 (m, 2H), 2.10 (d, 2H), 1.96-1.82 (m, 3H), 1.74-1.62 (m, IH), 1.6 1.40 (m, 9H).

Example 35: (/?)-l-[(3,5-Difluoro-phenylcarbamoyI)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-yV-(3,5-difluoro-phenyl)-acetamide

The sub-titled compound (2.02 g, colourless waxy solid) was prepared using a similai procedure to that used for the preparation of Example 38a but using 3,5-difluoroanilin place of 2,6-difluoroaniline.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.17 (s, IH), 7.16 (d, 2H), 6.63 (td, IH), 4.02 (s, 2H)

Example 35: (/?)-l-[(3,5-Difluoro-phenylcarbamoyl)-methyl]-3-(l-phenyl- i o cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

A mixture of 2-bromo-iV-(3,5-difluoro-phenyl)-acetamide (42 mg) (Example 35a) and phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example (50 mg) in MeCN (1 mL) was stirred at room temperature for 72h. The solid precipita

I S was filtered off, washed with ether and dried at 5O ⁰ C under vacuum overnight. The me liquors were evaporated, treated with ether and the resulting solid was filtered off. The solids were combined and crystallised from hot isopropyl alcohol to give the title compound (15 mg) as a colourless solid.

20 m/e 497 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 10.89 (s, IH), 7.33-7.17 (m, 7H), 7.01 (tt, IH), 5 5.06 (m, IH), 4.26-4.14 (m, 2H), 4.10-4.01 (m, IH), 3.63-3.49 (m, 4H), 3.43-3.32 (i

1 nc\ zπ /

Example 36: (/f)-l-[2-(4-methoxy-benzy!oxy)-ethyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane formate

^H

A mixture of l-(2-bromo-ethoxymethyl)-4-methoxy-benzene (42 mg) and 1-phenyl- cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 14e) (i mg) in MeCN (1 mL) was stirred at room temperature for 16h then heated to 8O ⁰ C unc nitrogen overnight. A further 1.1 equivalents of l-(2-bromo-ethoxymethyl)-4-methox> benzene were added and the mixture was heated at 8O ⁰ C for a further 48h. The volatile were evaporated and the crude product was purified by silica gel chromatography elut with 0-10% of a 10% cone. NHyMeOH solution in dichloromethane give 61 mg of an that was purified further by reverse phase HPLC (Gemini 5μM C6 phenyl column) ek with 5-95% MeOH/H ₂ O containing 0.1% formic acid over 30min with UV detection z 220nm. The relevant fractions were combined and evaporated to afford the title comp< (34 mg) as an oil.

m/e 492 [M] ⁺

¹ H NMR (400 MHz, CD ₃ OD): 5 8.48 (s, IH), 7.31-7.16 (m, 7H), 6.91-6.86 (m, 2H), 5.09-5.03 (m, IH), 4.44-4.39 (m, 2H), 3.92-3.63 (m, 6H), 3.46-3.29 (m, 5H), 2.99-2 (m, IH), 2.43-2.28 (m, 2H), 2.24-2.20 (m, IH), 2.15-1.86 (m, 4H), 1.75-1.51 (m, 1C

Example 37: (R)-l-(2-Phenethyloxy-ethyl)-3-(l-phenyl-cycloheptaπecarbon yloxy] azonia-bicyclo[2.2.2]octane bromide

H

The title compound (68 mg, 77%, gummy solid) was prepared according to a similar procedure to that used for Example 36 using [2-(2-bromo-ethoxy)-ethyl]-benzene in p of 1 -(2-bromo-ethoxymethyl)-4-methoxy-benzene.

m/e 476 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 7.32-7.11 (m, 10H), 5.00-4.92 (m, IH), 3.80-3.6! 3H), 3.60 (t, 2H), 3.41-3.11 (m, 6H), 2.99-2.88 (m, IH), 2.77 (t, 2H), 2.38-2.21 (m. 2H), 2.17-2.04 (m, 2H), 1.99-1.89 (m, IH), 1.84-1.67 (m, 2H), 1.67-1.32 (m, 10H).

Example 38: (R)-l-[(2,6-Difluoro-phenylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-N-(2,6-difluoro-phenyl)-acetamide

To a mixture of 2,6-difluoroaniline ( 1.12 g) and potassium carbonate (1.8 g) in 50 mL dichloromethane was added bromoacetyl bromide (1.5 mL) and the mixture was stirrer room temperature for 17h. Water was added and the mixture stirred for several hours t the phases were separated on a hydrophobic frit and the organic layer was evaporated, crude product was purified by silica gel chromatography eluting with 0-100% EtOAc/cyclohexane to give the sub-titled compound (0.92 g) as a white solid.

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.74 (s, IH), 7.31-7.21 (m, IH), 6.98 (t, 2H), 4.08 (s, 2

Example 38: (R)-l-[(2,6-Difluoro-phenylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

H

A mixture of 2-bromo-/V-(2,6-difIuoro-phenyl)-acetamide (Example 38a) (42 mg) and phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example (50 mg) in MeCN (1 mL) was stirred at room temperature for 19h. A further 20 mg 2- bromo-./V-(2,6-difluoro-phenyl)-acetamide was added and stirring continued for anothe 22h. The volatiles were evaporated and the residue was purified by silica gel chromatography eluting with 0-10% MeOH/dichloromethane to give the title compoui

10 (41 mg) as a colourless foam.

m/e 497 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 10.40 (s, IH), 7.42-7.35 (m, IH), 7.31-7.25 (m, 4 7.22-7.16 (m, 3H), 5.12-5.05 (m, IH), 4.32-4.27 (m, 2H), 4.10-4.02 (m, IH), 3.66-3

I ₅ (m, 4H), 3.43-3.31 (m, IH), 2.36-2.21 (m, 2H), 2.17-2.06 (m, 2H), 1.99-1.85 (m, 3t 1.77-1.66 (m, IH), 1.68-1.25 (m, 9H).

Example 39: (/?)-l-[(Methyl-phenyl-carbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane formate

20 a) 2-Bromo-/V-methyl-/V-phenyl-acetamide

To a mixture of iV-methyl aniline (5 mL) and potassium carbonate (9.6 g) in 100 mL

concentrated. The crude product was purified by silica gel chromatography eluting wi 100% EtOAc/cyclohexane. The relevant fractions were combined and evaporated and residue was taken up in dichloromethane and washed with water. The organic layer w, stirred with another portion of water for a few minutes and then the layers were separ∑ Evaporation of the organic layer gave the sub-titled compound (10.2 g) as a straw cok solid.

¹ H NMR (400 MHz, CDCl ₃ ): δ 7.50-7.36 (m, 3H), 7.31-7.26 (m, 2H), 3.67 (s, 2H), 3. (s, 3H).

Example 39: (/?)-l-[(Methyl-phenyl-carbamoyI)-methyl]-3-(l-phenyI- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane formate

H

A mixture of 2-bromo-./V-methyl-./V-phenyl-acetamide ((Example 39a) (38 mg) and 1- phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example (50 mg) in 1 mL MeCN was stirred at room temperature for 48h. The volatiles were evaporated and the residue purified by silica gel chromatography eluting with 0-10% MeOH/dichloromethane. Further purification by reverse phase HPLC eluting with 20- MeCN/H ₂ 0 containing 0.1% formic acid gave the title compound (17 mg) as a colour gum.

m/e 475 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ with a drop of TFA-D): δ 8.08 (s, IH), 7.47 (t, 2H), ^' (dd, 3H), 7.32-7.23 (m, 4H), 7.20-7.15 (m, IH), 5.09-4.98 (m, IH), 4.01-3.83 (m, 3 3.61-3.34 (m, 5H), 3.16-3.09 (s, 3H) 2.37-2.21 (m, 2H), 2.16-2.05 (m, 2H), 1.96-1.7

Example 40: (R)-l-[3-(4-Cyano-phenoxy)-propyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

H

A mixture of 4-(3-bromo-propoxy)-benzonitrile (41 mg) and 1-phenyl- cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 14e) (f mg) in MeCN (1 mL) was stirred at room temperature for 16h then heated to 8O ⁰ C unc nitrogen for a further 16h. The volatiles were evaporated and the residue was trituratec with ether to give an off-white solid that was triturated with EtOAc and dried under vacuum to give the title compound (61 mg) as a white solid.

m/e 487 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 7.78-7.73 (m, 2H), 7.33-7.26 (m, 4H), 1.22-1 Al IH), 7.09-7.04 (m, 2H), 5.05-4.99 (m, IH), 4.08 (t, 2H), 3.82 (ddd, IH), 3.43-3.26 5H), 3.21-3.08 (m, IH), 3.02-2.90 (m, IH), 2.39-2.22 (m, 2H), 2.17-2.00 (m, 4H), 1.78 (m, 3H), 1.68-1.43 (m, 10H).

Example 41: (R)-l-[(2,5-Difluoro-phenylcarbamoyI)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicydo[2.2.2]octane bromide

a) 2-Bromo-/V-(2,5-difluoro-phenyl)-acetamide

The sub-titled compound (3.6g, 90%, orange solid) was prepared by a similar procedi that used to prepare Example 38a using 2,5-difluoroaniline in place of 2,6-difluoroani

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.42 (s, IH), 8.13 (ddd, IH), 7.08 (ddd, IH), 6.82-6. s (m, IH), 4.04 (s, 2H).

Example 41: (R)-l-[(2,5-Difluoro-phenylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)- l-azonia-bicyclo[2.2.2]octane bromide

A mixture of 2-bromo-iV-(2,5-difluoro-phenyl)-acetamide (Example 41a) (47 mg) and phenyl-cycloheptanecarboxylic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Exampk (56 mg) in MeCN (1 mL) was stirred at room temperature for 30h. The resulting precipitate was filtered off, washed with ether and dried at 5O ⁰ C under vacuum overni to afford the title compound (65 mg) as a colourless solid.

m/e 497 [M] ⁺

¹ H NMR (400 MHz, DMSO-dD): δ 10.51 (s, IH), 7.82-7.75 (m, IH), 7.38-7.26 (m, . 7.21-7.16 (m, IH), 7.09-7.01 (m, IH), 5.12-5.06 (m, IH), 4.25 (s, 2H), 4.05 (ddd, 1 3.61-3.48 (m, 4H), 3.42-3.29 (m, IH), 2.36-2.23 (m, 2H), 2.17-2.06 (m, 2H), 1.98- 1 (m, 3H), 1.76-1.67 (m, IH), 1.66- 1.41 (m, 9H).

Example 42: (R)-l-[2-(4-Cyano-benzyloxy)-ethyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

H

A mixture of 4-(2-bromo-ethoxymethyl)-benzonitrile (41 mg, 0.17mmol) and 1-phen} cycloheptanecarboxylic acid (R)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 14e) (.' mg) in MeCN (1 mL) was stirred at room temperature for 16h then heated to 8O ⁰ C un< nitrogen overnight. The resulting solid was filtered off, washed with ether and dried u vacuum to give the title compound (70 mg) as a white solid.

m/e 487 [M] ⁺

H NMR (400 MHz, DMSO-D ₆ ): δ 7.80 (dd, 2H), 7.48 (d, 2H), 7.31-7.22 (m, 4H), 7.16 (m, IH), 5.06-4.96 (m, IH), 4.57 (s, 2H), 3.94-3.78 (m, 3H), 3.56-3.35 (m, 5H 3.27-3.18 (m, IH), 3.11-3.00 (m, IH), 2.36-2.18 (m, 2H), 2.08 (d, 2H), 1.98-1.77 (r 3H), 1.72-1.60 (m, IH), 1.60-1.38 (m, 9H).

Example 43: (/?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-[(6-trifluoromet hyI- pyridin-2-yIcarbamoyl)-methyl]-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-j¥-(3-trifluoromethyl-phenyl)-acetamide

The sub-titled compound (l.lg, quant., white solid) was prepared by a similar procedi that used to prepare Example 14f using 2-amino-6-(trifluoromethyl)pyridine in place c amino-pyridine and adding the chloroacetyl chloride at room temperature rather than < O ⁰ C.

Example 43: (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-[(6-trifluorometh yl- pyridin-2-ylcarbamoyl)-methyl]-l-azonia-bicyclo[2.2.2]octane chloride

H

The title compound (66 mg, 76%, white solid) was prepared by a similar procedure to used to prepare Example 14 using 2-chloro-iV-(3-trifluoromethyl-phenyl)-acetamide (Example 43a) in place of 2-chloro-/V-pyridin-2-yl-acetamide.

m/e 530 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 11.48 (s, IH), 8.25 (d, IH), 8.14 (t, IH), 7.67 (d. 7.33-7.26 (m, 4H), 7.22-7.17 (m, IH), 5.12-5.05 (m, IH), 4.28 (s, 2H), 4.11-4.03 (π IH), 3.66-3.49 (m, 4H), 3.41-3.29 (m, IH), 2.36-2.23 (m, 2H), 2.18-2.06 (m, 2H), 1.82 (m, 3H), 1.79-1.66 (m, IH), 1.66-1.40 (m, 9H).

Example 44: (/?)-l-[(4-Methyl-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phenyl - cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-./V-(4-methyl-pyridin-2-yl)-acetamide

The sub-titled compound (1.3g, 74%, solid) was prepared by a similar procedure to th used to prepare Example 14f using 2-amino-4-methylpyridine in place of 2-amino-pyi and adding the chloroacetyl chloride at room temperature rather than at O ⁰ C.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.77 (s, IH), 8.17 (d, IH), 8.03 (s, IH), 6.93 (d, IH), (s, 2H), 2.39 (s, 3H).

Example 44: (R)-l-[(4-Methyl-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phenyI- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

To a solution of 1-phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3 ester (Example 14e) (100 mg) in 1 mL MeCN was added 2-chloro-./V-(4-methyl-pyridi yl)-acetamide (Example 44a) (62 mg) and the mixture was stirred at room temperature 2 Ih. The precipitate was filtered off and dried under vacuum at 5O ⁰ C to give the title compound (80 mg) as a white solid.

m/e 476 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 10.98 (s, IH), 8.18 (d, IH), 7.85 (s, IH), 7.33-7. (m, 4H), 7.22-7.17 (m, IH), 7.02-7.00 (m, IH), 5.1 1-5.04 (m, IH), 4.24 (s, 2H), 4. 4.02 (m, IH), 3.64-3.50 (m, 4H), 3.40-3.29 (m, IH), 2.42-2.20 (m, 5H), 2.17-2.06 ( 2H), 1.98-1.84 (m, 3H), 1.79-1.67 (m, IH), 1.73-1.31 (m, 9H).

Example 45: (R)-l-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-/V-(5-chloro-pyridin-2-yl)-acetamide

The sub-titled compound (0.33g, 20%) was prepared by a similar procedure to that usi prepare Example 14f using 2-amino-5-chloropyridine in place of 2-amino-pyridine an adding the chloroacetyl chloride at room temperature rather than at 0 ⁰ C. 1H NMR (400 MHz, CDCl ₃ ): δ 8.80 (s, IH), 8.28 (d, IH), 8.19 (d, IH), 7.70 (dd, IH 4.20 (d, 2H).

Example 45: (/?)-l-[(5-Chloro-pyridin-2-ylcarbamoyl)-methyI]-3-(l-phenyl - cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

H The title compound (103 mg, 63%, white solid) was prepared by a similar procedure ti used to prepare Example 44 using 2-chloro-./V-(5-chloro-pyridin-2-yl)-acetamide (Exai 45a) in place of 2-chloro-/V-(4-methyl-pyridin-2-yl)-acetamide.

m/e 496 [M] ⁺ 1H NMR (400 MHz, DMSO-D ₆ ): δ 1 1.26 (s, IH), 8.41-8.39 (m, IH), 8.02-7.94 (m, 2 7.32-7.25 (m, 4H), 7.22-7.17 (m, IH), 5.12-5.03 (m, IH), 4.27 (s, 2H), 4.09-4.02 (π IH), 3.63-3.50 (m, 4H), 3.42-3.32 (m, IH), 2.37-2.23 (m, 2H), 2.17-2.05 (m, 2H), 1 1.84 (m, 3H), 1.78-1.65 (m, IH), 1.65-1.39 (m, 9H).

Example 46: (/?)-3-(l-Phenyl-cycloheptanecarbonyIoxy)-l-(/Molylcarbamoyl -met l-azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-iV-p-tolyl-acetamide

To a solution of p-toluidine (2.35 g) in 100 mL dichloromethane was added potassiurr carbonate (6.21 g). The reaction mixture was flushed with argon then bromoacetyl brc (1.6 mL) was added drop wise and then the reaction mixture was stirred for 17h. Watei added and the layers were separated. The organic layer was treated with cyclohexane < the volume was reduced in vacuo causing a solid to precipitate which was filtered off afford the sub-titled compound (2.67 g) as an off-white solid.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.06 (s, IH), 7.40 (d, 2H), 7.16 (d, 2H), 4.01 (s, 2H), (s, 3H).

Example 46: (/?)-3-(l-Phenyl-cycloheptanecarbonyIoxy)-l-(p-tolylcarbamoy l-met l-azonia-bicyclo[2.2.2]octane bromide

H

To 1-phenyl-cycloheptanecarboxylic acid (./?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 14e) (51 mg) in acetonitrile (1 mL) was added 2-bromo-N-/?-tolyl-acetamidc (Example 46a) (39 mg). The reaction was stirred at room temperature overnight and tr acetonitrile was removed under reduced pressure. The material was recrystallised froπ acetonitrile/ethyl acetate to afford the title compound as a colourless solid (16 mg).

m/e 475 [M] ⁺

¹ H NMR (400MHz, DMSO-D ₆ ): δ 10.40 (s, IH), 7.40 (d, 2H), 7.33-7.25 (m, 4H), 7.21 7.16 (m, IH), 7.13 (d, 2H), 5.1 1-5.04 (m, IH), 4.13 (q, 2H), 4.09-4.00 (m, IH), 3.64-3 (m, 4H), 3.42-3.36 (m, IH), 2.37-2.23 (m, 2H), 2.23 (s, 3H), 2.17-2.04 (m, 2H), 1.95-1 (m, 3H), 1.78-1.66 (m, IH), 1.65-1.40 (m, 9H).

Example 47: (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(/n-tolylcarbamoy l-

a) 2-Bromo-N-m-tolyl-acetamide

To a solution of m-toluidine (5.35 g) in 150 niL dichloromethane was added potassiui carbonate (17.3 g). The reaction mixture was flushed with argon then bromoacetyl bπ (3.6 mL) was added dropwise over ~ 15 mins and the reaction mixture was stirred for Water was added and the layers were separated. The organic layer was evaporated am residue was treated with EtOAc/cyclohexane. The precipitate was filtered off and discarded. The mother liquors were evaporated and purified by silica gel chromatogra eluting with 0-100% EtOAc/cyclohexane to give the sub-titled compound (6.13 g) as off-white solid.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.06 (s, IH), 7.38-7.28 (m, 2H), 7.27-7.21 (t, IH), 6.9 IH), 4.02 (s, 2H), 2.36 (s, 3H).

Example 47: (/?)-3-(l-Phenyl-cycIoheptanecarbonyloxy)-l-(/n-tolylcarbamo yl- methyl)-l-azonia-bicyclo[2.2.2]octane bromide

H

To 1-phenyl-cycloheptanecarboxylic acid (R)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 14e) (50 mg) in acetonitrile ( 1 mL) was added 2-bromo-N-m-tolyl-acetamic (Example 47a) (38 mg). The reaction was stirred at room temperature for 26 hours am acetonitrile was removed under reduced pressure. The material was purified by silica chromatography eluting with 0-10% MeOH/dichloromethane to afford the title compc

CVl mσλ ϊIQ a rnlniirlpcc fnqm

m/e 475 [M] ⁺

¹ H NMR (400MHz, DMSO-D ₆ ): δ 10.40 (s, IH), 7.39 (s, IH), 7.32-7.25 (m, 5H), 7.Z 7.15 (m, 2H), 6.92 (d, IH), 5.11-5.05 (m, IH), 4.14 (q, 2H), 4.11-4.01 (m, IH), 3.64-2 (m, 4H), 3.42-3.30 (m, IH), 2.39-2.21 (m, 5H), 2.18-2.05 (m, 2H), 1.97-1.84 (m, 3H). 1.77-1.66 (m, IH), 1.65-1.39 (m, 9H).

Example 48: (/?)-l-(Oxazol-2-ylcarbamoylmethyl)-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-N-oxazol-2-yl-acetamide

A solution of bromoacetyl bromide (0.44 mL) in dry CHCl ₃ (5 niL) was added dropwi a suspension of 2-amino-l,3-oxazole (0.39 g) and triethylamine (0.96 mL) in dry CHC (92 mL) at room temperature. The brown mixture was allowed to stir for 16h, then quenched with H ₂ O (2 mL) and stirred for 20mins before concentrating under reduced pressure to a light brown solid. The crude product was purified by silica gel chromatography eluting with 1-3% MeOH/dichloromethane to give the title compoun< (0.56g) as an off-white solid.

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 11.61 (s, IH), 7.89 (s, IH), 7.12 (s, IH), 4.11 (s,

Example 48: (R)-l-(Oxazol-2-ylcarbamoylmethyI)-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

H

1-Phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Exam 14e) (0.31 mmol) and 2-bromo-iV-oxazol-2-yl-acetamide (Example 48a) (0.31 mmol) stirred in anhydrous MeCN at room temperature for 18 hours. The reaction mixture v concentrated in vacuo and the yellow solid purified by flash silica gel column chromatography eluting with 0-15% MeOH/dichloromethane to give the title compou (100 mg) as a white solid.

m/e 452 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ , 353K): δ 7.67 (s, IH), 7.35-7.29 (m, 4H), 7.26-7.18 IH), 7.02 (s, IH), 5.14-5.05 (m, IH), 4.17-4.04 (m, 3H), 3.66-3.56 (m, 4H), 3.52-3 (m, IH), 2.42-2.29 (m, 2H), 2.24-2.12 (m, 2H), 2.10-1.86 (m, 3H), 1.82-1.70 (m, Hi 1.70-1.47 (m, 9H).

Example 49: (/?)-l-[(6-Methyl-pyridazin-3-yIcarbamoyl)-methyl]-3-(l-phen yl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-/V-(6-methyl-pyridazin-3-yl)-acetamide

A solution of bromoacetyl bromide (0.22 mL) in dry CHCl ₃ (4 mL) was added slowly suspension of 3-amino-6-methylpyridazine (0.24 g) and triethylamine (0.47 mL) in dv CHCl ₃ (45 mL) at room temperature. The brown mixture was allowed to stir for 3.5hr then quenched with H ₂ O (1.5 mL) and stirred for 20mins before concentrating under reduced pressure to a brown solid. The crude product was purified by silica gel chromatography eluting with 1-2% MeOH/dichloromethane. The relevant fractions w combined and evaporated to give the title compound (0.20 g) as a pinkish/beige solid.

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 11.41 (s, IH), 8.18 (d, IH), 7.59 (d, IH), 4.17 (s, ; 2.57 (s, 3H).

Example 49: (/?)-l-[(6-Methyl-pyridazin-3-yIcarbamoyI)-methyI]-3-(l-phen yl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane bromide

H

1-Phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Exam] 14e) (0.20 mmol) and 2-bromo-yV-(6-methyl-pyridazin-3-yl)-acetamide (Example 49a (0.20 mmol) were stirred together in anhydrous MeCN at room temperature for 18 hoi The reaction mixture was concentrated in vacuo and the yellow solid purified by flash silica gel column chromatography eluting with 0-15% MeOH/dichloromethane to giv( title compound (65 mg) as a tan solid.

m/e 477 [M] ⁺

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.19 (d, IH), 7.38 (d, IH), 7.27 (d, 4H), 7.20-7.12 (r IH), 5.18-4.96 (m, 3H), 4.41 (dd, IH), 4.1 1-3.95 (m, 3H), 3.81 (d, IH), 3.47-3.37 ( IH), 2.66 (s, 3H), 2.45-2.27 (m, 2H), 2.26-2.13 (m, 2H), 2.08-1.96 (m, 3H), 1.81-1 (m, IH), 1.69-1.30 (m, 9H).

Example 50: (/?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrimidin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-/V-pyrimidin-2-yl-acetamide

A solution of 2 amino-pyrimidine (2.0 g) in dry dichloromethane (17 mL) under nitroj at O ⁰ C was treated with triethylamine (2.6 mL) followed by slow addition of chloroaα r-hlnriHp ( \ ^ ml 1 8 immnl^ Thp rpnrtinn miγtiirp WSQ nllnwpri tn warm πn tn rnnrn

The phases were separated and the aqueous layer was extracted with dichloromethane The combined organic layers were washed with brine, dried over sodium sulphate, fill and concentrated to give the crude product which was purified by silica gel chromatography eluting with 10% MeOH/dichloromethane. The relevant fractions we 5 combined and evaporated to give the title compound (1.20 g) as a green solid.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.84 (s, IH), 8.65 (d, 2H), 7.09 (t, IH), 4.46 (s, 2H).

Example 50: (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrimidin-2-o ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride

H

1-Phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Exam] 14e, 0.30 mmol) and 2-chloro-/V-pyrimidin-2-yl-acetamide (Example 50a) (0.36 mmo MeCN (1.5 mL) were stirred together at room temperature overnight. The reactions mixture was concentrated in vacuo and the residue purified by silica gel chromatograp eluting with 0-10% MeOH/dichloromethane to give the title compound as a white soli mg).

m/e 463 [M] ⁺ o ¹ H NMR (400 MHz, DMSO-D ₆ ): δ 1 1.19 (s, IH), 8.66 (d, 2H), 7.32-7.24 (m, 4H), 7 7.14 (m, 2H), 5.09-5.03 (m, IH), 4.48 (s, 2H), 4.06 (ddd, IH), 3.65-3.51 (m, 4H), 2 3.34 (m, IH), 2.37-2.21 (m, 2H), 2.15-2.07 (m, 2H), 1.96-1.80 (m, 3H), 1.75-1.64 (, IH), 1.63-1.38 (m, 9H). ₅ Example 51: (/f)-l-[(5-Cyano-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-./V-(5-cyano-pyridin-2-yl)-acetamide

A solution of 2-amino-5-cyano pyridine (2.0 g) in dry dichloromethane (17 mL) unda nitrogen at O ⁰ C was treated with triethylamine (2.6 mL) followed by slow addition of chloroacetyl chloride (1.5 mL). The reaction mixture was allowed to warm up to roorr temperature. After 2h, the mixture was partitioned between dichloromethane and wate The phases were separated and the aqueous layer was extracted with dichloromethane The combined organic layer was washed with brine, dried over sodium sulphate, filter and concentrated to give the crude product which was purified by silica gel chromatography eluting with 50% EtOAc/cyclohexane. The relevant fractions were combined and evaporated to give the title compound (2.17g) as a light brown solid.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.99 (s, IH), 8.61 (dd, IH), 8.36 (dd, IH), 8.00-7.97 IH), 4.23 (s, 2H).

Example 51: (i?)-l-[(5-Cyano-pyridin-2-yIcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)- 1 -azonia-bicyclo[2.2.2]octane chloride

1 -Phenyl-cycloheptanecarboxylic acid (R)-( 1 -aza-bicyclo[2.2.2]oct-3-yl) ester (Examj 14e) (0.30 mmol) and 2-chloro-N-(5-cyano-pyridin-2-yl)-acetamide (Example 51a) (0. mmol) in MeCN (1.5 mL) were stirred together at room temperature overnight. The reaction mixture was concentrated in vacuo and the residue purified by silica gel

m/e 487 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 1 1.56 (s, IH), 8.81 (dd, IH), 8.31 (dd, IH), 8.09 IH), 7.32-7.25 (m, 4H), 7.23-7.17 (m, IH), 5.11-5.04 (m, IH), 4.32 (s, 2H), 4.10-4 (m, IH), 3.63-3.50 (m, 4H), 3.42-3.29 (m, IH), 2.37-2.23 (m, 2H), 2.17-2.05 (m, 2] 2.00-1.82 (m, 3H), 1.78-1.65 (m, IH), 1.65-1.40 (m, 9H).

Example 52: (R)-3-(l-Phenyl-cycloheptanecarbonyIoxy)-l-(pyrimidin-5- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-ChloiO-/V-pyrimidin-5-yl-acetamide

5-Aminopyrimidine (450 mg) was suspended in DCE (2 mL) and acetonitrile (2 mL) : microwave vial. Chloroacetyl chloride (0.377 mL) was added with stirring. The vial \ sealed and the reaction mixture was heated in the microwave at 80 ⁰ C for 5 minutes. T solid was filtered off, washed with acetonitrile (2 x 5 mL), DCE (2 x 5 mL) and penta x 30 mL) and then partioned between saturated sodium bicabonate and DCE (50 mL/* mL ) ensuring the aqueous layer was still basic. The organic layer was separated and t aqueous layer was extracted with DCE (2 x 75 mL). The combined organic layer was over magnesium sulfate and evaporated to give the sub-titled compound (200 mg) as i yellow solid.

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 10.71 (s, IH), 9.00 (s, 2H), 8.93 (s, IH), 4.35 (s, _ ^■

Example 52: (R)-3-(l-PhenyI-cycloheptanecarbonyloxy)-l-(pyrimidin-5- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride

H

1-Phenyl-cycloheptanecarboxylic acid (7?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Exam] 14e) (115 mg) in acetonitrile (2 mL) was treated with 2-chloro-/V-pyrimidin-5-yl-acet; (Example 52a) (66 mg) to give a dark brown solution that was stirred at room tempeπ overnight. The resulting solid was filtered off, washed with cold acetonitrile (2 mL) a pentane (3 mL) and dried under vacuum at 45 ⁰ C to give the title compound as a tan sc (151 mg).

m/e 463 [M] ⁺ 1H NMR (400 MHz, DMSO-D ₆ ): δ 11.70 (s, IH), 9.01 (s, 2H), 8.93 (s, IH), 7.33-7.: (m, 4H), 7.22-7.16 (m, IH), 5.11-5.05 (m, IH), 4.34 (s, 2H), 4.12-4.04 (m, IH), 3.< 3.52 (m, 4H), 3.43-3.31 (m, IH), 2.37-2.22 (m, 2H), 2.17-2.06 (m, 2H), 1.99-1.83 ( 3H), 1.79-1.67 (m, IH), 1.65-1.40 (m, 9H).

Example 53: (/?)-l-[(3-Fluoro-pyridin-2-ylcarbamoyl)-methyI]-3-(l-phenyl - cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-/V-(3-fluoro-pyridin-2-yl)-acetamide

2-Amino-3-fluoropyridine (1.5 g) was dissolved in DCE (15 mL) and chloroacetylchlc (1.1 mL) was added dropwise. The reaction was heated in a microwave at 8O ⁰ C for 5rr The reaction mixture was cooled and the resulting solid was filtered off, washed with MeCN and pentane then suspended in dichloromethane and aqueous NaHCO ₃ (sat) w< _n JJ _λ J T ¹ U,-

concentrated. The crude product was purified by silica gel chromatography eluting wi 100% EtOAc/cyclohexane to give the title compound as a white solid (800 mg).

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 10.61 (s, IH), 8.27 (dt, IH), 7.83-7.77 (m, IH), 1 7.35 (m, IH), 4.37 (s, 2H).

Example 53: (R)-l-[(3-Fluoro-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

H 1-Phenyl-cycloheptanecarboxylic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Exam] 14e) (0.30 mmol) and 2-chloro-/V-(3-fluoro-pyridin-2-yl)-acetamide (Example 53a) (0 mmol) in MeCN (1.5 mL) were stirred at room temperature overnight. The reaction mixture was concentrated in vacuo and the residue purified by silica gel chromatograp eluting with 0-10% MeOH/dichloromethane to give the title compound as a white soli mg).

m/e 480 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 1 1.07 (s, IH), 8.25 (dt, IH), 7.81 (ddd, IH), 7.3S (ddd, IH), 7.31-7.24 (m, 4H), 7.22-7.15 (m, IH), 5.1 1-5.04 (m, IH), 4.36 (s, 2H), <■ 4.04 (m, IH), 3.66-3.52 (m, 4H), 3.44-3.32 (m, IH), 2.37-2.21 (m, 2H), 2.19-2.06 (> 2H), 1.99-1.82 (m, 3H), 1.77-1.65 (m, IH), 1.65-1.38 (m, 9H).

Example 54: (/?)-l-[(3-Fluoro-pyridin-4-ylcarbamoyl)-methyl]-3-(l-phenyl - cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-./V-(3-fluoro-pyridin-4-yl)-acetamide

A solution of 3-fluoro-pyridin-4-ylamine (0.2 g) in dry dichloromethane (2 mL) unde nitrogen at O ⁰ C was treated with triethylamine (0.28 mL) followed by slow addition o chloroacetyl chloride (0.16 mL). The reaction mixture was allowed to warm up to roo temperature. After 2h, the mixture was partitioned between dichloromethane and wat( The phases were separated and the aqueous layer was extracted with dichloromethane The combined organic layer was washed with brine, dried over sodium sulphate, filtej and concentrated to give the crude product which was purified by silica gel chromatography eluting with 0-100% EtOAc/cyclohexane. The relevant fractions wei combined and evaporated to give the title compound (0.1 Ig) as a pink solid.

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 10.55 (s, IH), 8.56 (d, IH), 8.35 (d, IH), 8.16 (d IH), 4.44 (s, 2H).

Example 54: (/?)-l-[(3-Fluoro-pyridin-4-ylcarbamoyl)-methyI]-3-(l-phenyl - cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

H

1-Phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Exam] 14e) (0.30 mmol) and 2-chloro-./V-(3-fluoro-pyridin-4-yl)-acetamide (Example 54a) (( mmol) in MeCN (1.5 mL) were stirred together at room temperature overnight. The reaction mixture was concentrated in vacuo and the residue purified by silica gel chromatography eluting with 0-10% MeOH/dichloromethane to give the title compou a white solid (1 10 mg).

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 10.92 (s, IH), 8.56 (d, IH), 8.35 (d, IH), 8.03 (c IH), 7.32-7.26 (m, 4H), 7.21-7.16 (m, IH), 5.11-5.05 (m, IH), 4.41-4.29 (m, 2H), 4.02 (m, IH), 3.63-3.50 (m, 4H), 3.42-3.30 (m, IH), 2.36-2.23 (m, 2H), 2.17-2.06 ( 2H), 1.98-1.82 (m, 3H), 1.78-1.65 (m, IH), 1.66-1.40 (m, 9H).

Example 55: (/?)-3-(l-Phenyl-cycloheptanecarbonyIoxy)-l-{2-[(pyrazine-2- carboi amino]-ethyl}-l-azonia-bicyclo[2.2.2]octane bromide

a) Pyrazine-2-carboxylic acid (2-bromo-ethyl)-amide

2-Pyrazine carboxylic acid (1 g) in dichloromethane (30 mL) was treated with triethylamine (1.27 mL) and HATU (3.6 g) and the mixture was stirred for 10 minutes solution of 2-bromoethylamine hydrobromide (1.5 g) and triethylamine (1.27 mL) in dichloromethane (20 mL) was added and the reaction mixture was stirred for 3 hours. Water (50 mL) was added and the organic layer was separated and washed with water 50 mL). The organic layer was dried over magnesium sulphate and evaporated to giv< crude product which was purified by silica gel chromatography eluting with 0-100% EtOAc/dichloromethane. The relevant fractions were combined and evaporated to giv residue which was dissolved in EtOAc (40 mL) and washed with saturated sodium hydrogen carbonate ensuring the aqueous layer was basic. The organic layer was driec over magnesium sulfate and evaporated to give the sub-titled compound (1.0 g) as a w solid.

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 9.21 (d, IH), 9.14 (t, IH), 8.90 (d, IH), 8.75 (dd, 3.75-3.69 (m, 2H), 3.66-3.60 (m, 2H).

Example 55: (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-{2-[(pyrazine-2-c arboi amino]-ethyl}-l-azonia-bicyclo[2.2.2]octane bromide

H

Pyrazine-2-carboxylic acid (2-bromo-ethyl)-amide (Example 55a) (87 mg) was added solution of 1-phenyl-cycloheptanecarboxylic acid (R)-( l-aza-bicyclo[2.2.2]oct-3-yl) e (Example 14e) (113 mg) in acetonitrile (2 mL). The reaction mixture was allowed to room temperature for 16h. A solid precipitated out and was filtered off, washed with i acetontrile and dried under vacuum at 40 ⁰ C to give the title compound (96 mg) as a w solid.

m/e 477 [M] ⁺ 1H NMR (400 MHz, DMSO-D ₆ ): δ 9.22 (t, IH), 9.17 (d, IH), 8.88 (d, IH), 8.72 (dd IH), 7.33-7.25 (m, 4H), 7.22-7.16 (m, IH), 5.01-4.96 (m, IH), 3.89 (ddd, IH), 3.7: 3.57 (m, 2H), 3.51-3.28 (m, 5H), 3.22 (dt, IH), 3.13-3.02 (m, IH), 2.38-2.30 (m, IP 2.30-2.20 (m, IH), 2.17- 2.06 (m, 2H), 1.97-1.76 (m, 3H), 1.69-1.37 (m, 10H).

Example 56: (/?)-3-(l-Phenyl-cycIoheptanecarbonyloxy)-l-([l,2,4]thiadiaz ol-5- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-yV-[ 1 ,2,4]thiadiazol-5-yl-acetamide

A solution of [l,2,4]-thiadiazol-5-ylamine (3.0 g) in dry dichloromethane (30 mL) un nitrogen at O ⁰ C was treated with triethylamine (4.6 mL) followed by slow addition of chloroacetyl chloride (2.6 mL). The reaction mixture was allowed to warm up to roorr temperature. After 2h, the mixture was partitioned between dichloromethane and wate The phases were separated and the aqueous layer was extracted with dichloromethane

and concentrated to give the crude product which was purified by silica gel chromatography eluting with 50-75% EtOAc/cyclohexane to give the title compound g) as a yellow solid.

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 13.32 (s, IH), 8.51 (s, IH), 4.52 (s, 2H).

Example 56: (R)-3-(l-Phenyl-cydoheptanecarbonyloxy)-l-([l,2,4]thiadiazol -5- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride

H 1-phenyl-cycloheptanecarboxylic acid (7?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Examp 14e) (0.30 mmol) and 2-chloro-/V-[l,2,4]thiadiazol-5-yl-acetamide (0.36 mmol) (Exan 56a) in MeCN (1.5 niL) were stirred at room temperature overnight. The reaction mix was filtered and the solid obtained was washed with cold MeCN to give the title comp (30 mg) as a solid.

m/e 469 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 13.70 (s, IH), 8.51 (s, IH), 7.34-7.27 (m, 4H), 7. 7.17 (m, IH), 5.11-5.05 (m, IH), 4.54-4.43 (m, 2H), 4.12-4.05 (m, IH), 3.67-3.53 (i 4H), 3.45-3.33 (m, IH), 2.38-2.24 (m, 2H), 2.18-2.05 (m, 2H), 1.99-1.83 (m, 3H), 1 1.67 (m, 1 H), 1.67- 1.40 (m, 9H).

Example 57: (R)-3-(l-PhenyI-cycIoheptanecarbonyloxy)-l-{3-[(pyridine-2-c arbon amino]-propyl}-l-azonia-bicyclo[2.2.2]octane bromide

a) Methanesulfonic acid 3-[(pyridine-2-carbonyl)-amino]-propyl ester

Isobutyl chloroformate (3.35 mL) was added to a solution of 2-pyridine carboxylic ac (2.10 g) and TV-methyl morpholine (2.82 mL) in dry THF (85 mL) at 0 ⁰ C. After 15m amino- 1-propanol (1.31 mL) was added and the mixture stirred overnight. The reacti< mixture was concentrated to a pink solid in vacuo and was passed through a pad of sil (1-5% MeOH/dichloromethane). The resultant brown oil was taken up in dichlorome (85 mL) and cooled to 0 ⁰ C. To this solution was added Et ₃ N (4.75 mL) and methane sulfonylchloride (2.0 mL). After 30 mins the reaction was warmed to room temperati and stirred for 2.5 h before quenching with H ₂ O (50 mL). The layers were separated ; the organic phase washed with sat. NaHCO _{3 (aq)} and dried (MgSO ₄ ). Concentration ur reduced pressure gave an orange oil which was purified by silica gel chromatography eluting with 90% EtOAc/cyclohexane to give the sub-titled compound (2.06 g) as an orange oil.

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.55 (ddd, IH), 8.29 (s, IH), 8.17 (dt, IH), 7.88-7.83 ( IH), 7.45 (ddd, IH), 4.35 (t, 2H), 3.63 (q, 2H), 3.07 (s, 3H), 2.1 1 (p, 2H).

b) Pyridine-2-carboxylic acid (3-bromo-propyl)-amide

A mixture of methanesulfonic acid 3-[(pyridine-2-carbonyl)-amino]-propyl ester (Exa 57a) (1.96 g) and lithium bromide (3.29 g) in acetone (19 mL) was heated to reflux fc h. The reaction mixture was cooled to room temperature, concentrated in vacuo and t residue partitioned between EtOAc/H ₂ O (60 mL, 1: 1). The phases were separated anc aqueous phase further extracted with EtOAc (2 x 25 mL). The combined organics we dried (MgSO ₄ ) and concentrated in vacuo to a brown oil which solidified on standing. Purification by silica gel chromatography eluting with 0-100% EtOAc/cyclohexane gi

¹ H NMR (400 MHz, CDCl ₃ ): δ 8.55 (ddd, IH), 8.25-8.12 (s, IH), 8.19 (dt, IH), 7.85 ( IH), 7.43 (ddd, IH), 3.64 (q, 2H), 3.50 (t, 2H), 2.22 (p, 2H).

Example 57: (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-{3-[(pyridine-2-c arboii amino]-propyl}-l-azonia-bicyclo[2.2.2]octane bromide

H

1-Phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Examt 14e) (0.31 mmol) and pyridine-2-carboxylic acid (3-bromo-propyl)-amide (Example 5 (0.31 mmol) were stirred together in anhydrous MeCN (3 mL) at room temperature fo days. The reaction mixture was concentrated in vacuo and the solid purified by silica chromatography eluting with 0-15% MeOH/dichloromethane to give the title compoui (145 mg) as a white solid.

m/e 490 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 8.94 (t, IH), 8.62 (ddd, IH), 8.03-7.95 (m, 2H), ' (ddd, IH), 7.31-7.24 (m, 4H), 7.19-7.13 (m, IH), 5.01-4.95 (m, IH), 3.76 (ddd, IH) 3.51-3.09 (m, 7H), 3.09-3.01 (m, IH), 2.93-2.82 (m, IH), 2.36-2.22 (m, 2H), 2.14-2 (m, 2H), 1.99-1.90 (m, IH), 1.92-1.71 (m, 4H), 1.68-1.39 (m, 10H).

Example 58: (R)-l-[(2-Methyl-pyrimidin-4-ylcarbamoyl)-methyl]-3-(l-pheny l- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-N-(2-methyl-pyrimidin-4-yl)-acetarnide

H

,ISL

Cl

2-Methyl-pyrimidin-4-yl amine (545 mg) was suspended in DCE (5 niL) and chloroacetylchloride (0.4 mL) was added dropwise. The reaction was heated in a microwave at 8O ⁰ C for 5mins. The reaction mixture was cooled to give a solid that wa fϊtered, washed with dichloromethane then suspended in dichloromethane and sat. NaHCO ₃ (aq) was added. The organic phase was collected and the aqueous layer extπ with dichloromethane (x2). The combined organic layer was dried over sodium sulphc and concentrated. The crude product was purified by silica gel chromatography elutinj with 0-10%MeOH/dichloromethane to give the sub-titled compound as a yellow solid mg, 7.5 %).

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 1 1.16 (s, IH), 8.58 (d, IH), 7.84 (d, IH), 4.37 (s, 2.53 (s, 3H).

Example 58: (R)-l-[(2-MethyI-pyrimidin-4-ylcarbamoyl)-methyl]-3-(l-pheny l- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

H

1-Phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) ester (Examr. 14e, 0.29 mmol) and 2-chloro-N-(2-methyl-pyrimidin-4-yl)-acetamide (Example 58a) (0.35 mmol) in MeCN (2.0 mL) were stirred together at room temperature for 16h. Tl reaction mixture was concentrated in vacuo and the residue purified by silica gel chromatography eluting with 0-10% MeOH/dichloromethane to give the title compoui a white solid (55 mg).

m/e 477 [M] ⁺ 1H νMR (400 MHz, DMSO-D ₆ ): δ 11.40 (s, IH), 8.61 (d, IH), 7.75 (d, IH), 7.32-7.: (m, 4H), 7.23-7.17 (m, IH), 5.10-5.03 (m, IH), 4.28 (s, 2H), 4.09-4.01 (m, IH), 3.6

3.48 (m, 4H), 3.40-3.30 (m, IH), 2.50 (s, 3H), 2.36-2.24 (m, 2H), 2.17-2.05 (m, 2E 1.98-1.84 (m, 3H), 1.78-1.65 (m, IH), 1.64-1.41 (m, 9H).

Example 59: (R)-l-[(6-Methyl-pyrimidin-4-yIcarbamoyI)-methyl]-3-(l-pheny l- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

a) 2-Chloro-iV-(6-methyl-pyrirnidin-4-yl)-acetamide

6-Methyl-pyrimidin-4-yl amine (545 mg) was suspended in DCE (5 mL) and chloroacetylchloride (0.4 mL) was added dropwise. The reaction was heated in a microwave at 8O ⁰ C for 5mins. The reaction mixture was cooled, filtered and a solid obtained. The reaction was repeated a second time and both batches of solid were combined, washed with dichloromethane then suspended in dichloromethane and sat. NaHCO ₃ (aq.) was added. The organic phase was collected and the aqueous layer extr with dichloromethane (x2). The combined organic layer was dried over sodium sulphc and concentrated. The crude product was purified by silica gel chromatography elutinj with 0-10%MeOH/dichloromethane to give the sub-titled compound as a yellow solid mg).

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 11.11 (s, IH), 8.76 (d, IH), 7.91 (s, IH), 4.38 (s, 2 2.44 (s, 3H).

Example 59: (/?)-l-[(6-Methyl-pyrimidin-4-ylcarbamoyl)-methyl]-3-(l-phen yl- cycloheptanecarbonyloxy)-l-azonia-bicyclo[2.2.2]octane chloride

1-Phenyl-cycloheptanecarboxylic acid (7?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Examj 14e) (0.30 mmol) and 2-chloro-7V-(6-methyl-pyrimidin-4-yl)-acetamide (Example 59a 5 (0.36 mmol) in MeCN (2. niL) were stirred together at room temperature for 16h. The reaction mixture was concentrated in vacuo and the residue purified by silica gel chromatography eluting with 0-10% MeOH/dichloromethane to give the title compoui a white solid (125 mg).

io m/e 477 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 11.39 (s, IH), 8.76 (d, IH), 7.83 (s, IH), 7.34-7.: (m, 4H), 7.22-7.17 (m, IH), 5.10-5.04 (m, IH), 4.32 (s, 2H), 4.10-4.01 (m, IH), 3.6 3.50 (m, 4H), 3.43-3.31 (m, IH), 2.42 (s, 3H), 2.37-2.23 (m, 2H), 2.18-2.06 (m, 2H; 1.98-1.81 (m, 3H), 1.78-1.66 (m, IH), 1.65-1.39 (m, 9H).

I ₅

Example 60: (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-{2-[(pyridine-2-c arbon amino]-ethyl}-l-azonia-bicyclo[2.2.2]octane bromide

a) Pyridine-2-carboxylic acid (2-bromo-ethyl)-amide

Picolinic acid (0.99 g) in dichloromethane (30 mL) was treated with triethylamine (1.2 mL) and HATU (3.6 g). The mixture was stirred for 10 minutes then a solution of 2- bromoethylamine hydrobromide (1.5 g) and triethylamine (1.27 mL) in dichloromethai

over magnesium sulphate and evaporated to afford the crude product which was purifi silica gel chromatography eluting with 0-100% EtOAc/dichloromethane. The relevant fractions were combined and evaporated, dissolved up in EtOAc (40 mL) and washed saturated sodium hydrogen carbonate solution, ensuring the aqueous layer remained b 5 The organic layer was dried over magnesium sulphate and evaporated to give the sub- compound (0.88 g) as a white solid.

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 9.01 (t, IH), 8.66 (ddd, IH), 8.07-7.98 (m, 2H), 7 (ddd, IH), 3.70 (q, 2H), 3.62 (t, 2H). 0

Example 60 : (R)-3-( 1 -Pheny I-cycloheptanecarbony loxy )- 1 - { 2- [(py ridine-2-carbor aminoJ-ethylJ-l-azonia-bicyclotZ.Z.lloctane bromide

H

Pyridine-2-carboxylic acid (2-bromo-ethyl)-amide (Example 60a) (75 mg) was added5 solution of 1-phenyl-cycloheptanecarboxylic acid (/?)-( l-aza-bicyclo[2.2.2]oct-3-yl) e: (Example 14e) (98 mg) in acetonitrile (2 mL). The reaction mixture was allowed to st room temperature for 16h. A further 10 mg of pyridine-2-carboxylic acid (2-bromo-etl amide was added and the reaction mixture was stirred for 8h. The volatiles were evaporated and the residue was purified by silica gel chromatography eluting with 0- 1o MeOH/dichloromethane to give the title compound (55 mg) as a white solid.

m/e 476 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 9.12 (t, IH), 8.64-8.62 (m, IH), 8.05-7.97 (m, 2fl 7.61 (ddd, IH), 7.32-7.25 (m, 4H), 7.22-7.15 (m, IH), 5.02-4.96 (m, IH), 3.88 (ddd5 IH), 3.71-3.55 (m, 2H), 3.49-3.27 (m, 5H), 3.22 (dt, IH), 3.12-3.02 (m, IH), 2.38-: (m, 2H), 2.17-2.07 (m, 2H), 1.96-1.75 (m, 3H), 1.69-1.38 (m, 10H).

Example 61: (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(3-pyridin-4-yl-p ropyl azonia-bicyclo[2.2.2]octane bromide

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

A solution of 3-pyridin-4-yl-propan-l-ol (2.88 mL) in hydrobromic acid (16 niL, 14 L- mmol) was heated at reflux at 135°C for 18h. The cooled solution was concentrated ur vacuum and the residue was re-dissolved in wopropanol and re-concentrated (this proc was repeated three more times). The residue was dissolved in /søpropanol, decolourise boiling with activated charcoal, filtered, and the clear solution left to crystallise in a fr< over 48h. The resulting crystals were removed by filtration, washed with ώøpropanol / diethyl ether (1: 1) followed by diethyl ether and then dried under vacuum at 40 ⁰ C and room temperature to afford the sub-titled compound as a pale brown solid (3.55 g).

¹ H NMR (400 MHz, D ₂ O): δ 8.64 (d, 2H), 7.96 (d, 2H), 3.52 (t, 2H), 3.12 (t, 2H), 2.31 (quint., 2H).

Example 61: (/?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(3-pyridin-4-yl- propyl) azonia-bicyclo[2.2.2]octane bromide

H

4-(3-Bromo-propyl)-pyridine hydrobromide(Example 61a) (0.210 g) was added to diet ether (10 mL) and sodium hydroxide solution (4 mL) (10%) in a separating funnel and mixture shaken and separated. The ethereal layer was washed with water (2 x 10 mL), dried (MgSO ₄ ) and evaporated to afford the free base as an oil. To the residue was add

(20 mL) gave an oil, the supernatant was removed by decantation and the residue was with ethyl acetate (2 x 20 mL). The oil was crystallised by stirring with diethyl ether mL) and the solid washed twice with diethyl ether (2 x 20 mL) to afford the titled compound as a solid (0.094 g).

m/e 447 [M] ⁺

¹ H NMR (400 MHz, DMSO-D ₆ ): δ 8.51 (d, 2H), 7.35 - 7.30 (m, 4H), 7.30 - 7.27 (m, : 7.24 - 7.18 (m, IH), 5.07 - 5.01 (m, IH), 3.81 (ddd, IH), 3.43 - 3.27 (m, 2H), 3.21 - 3 (m, IH), 3.10 (d, IH), 2.97 - 2.88 (m, 2H), 2.59 (t, 2H), 2.40 - 2.27 (m, 3H), 2.18 - 2.1 (m, 2H), 2.04 - 1.76 (m, 5H), 1.72 - 1.43 (m, 10H).

Pharmacological Analysis M3 Receptor Activity Assay The affinity (pICso) of compounds to the M ₃ receptor was determined by competition binding of [ ³ H]N-methyl scopolamine (NMS) to CHO-Kl (Chinese Hamster Ovary) c membranes expressing the human muscarinic acetylcholine M ₃ receptor (M ₃ -ACh) in scintillation proximity assay (SPA) format.

SPA beads were precoated with membranes and then incubated at 2 mg of beads per v with with serial dilutions of the compounds of the invention, [ ³ H]NMS at 0.2nM, half (experimentally determined dissociation constant) and assay buffer (20 mM HEPES p containing 5 mM MgCl ₂ ). The assay was conducted in a final volume of 200 μL, in tr presence of 1% (v/v) dimethyl sulphoxide (DMSO). Total binding of [ ³ H]NMS was determined in the absence of competing compound and non-specific binding of [ ³ H]N was determined in the presence of 1 μM atropine. The plates were incubated for 16 h( at room temperature and then read on Wallac Microbeta"' using a normalised ³ H protc The pICso, defined as the negative logarithm of the concentration of compound require 50% reduction in specific [ ³ H]-NMS binding, was determined. Table 1 shows the pIC figures for some representative Examples.

Table 1

Table 2 gives IC ₅₀ strengths for the compounds of the examples. Table 2

M3 Binding IC ₅₀ <2nM "+++"; IC ₅₀ 2-1OnM "++"; IC ₅₀ > 1OnM "+"; NT - Not Testec

Measurement of Plasma Protein Binding

5 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 tl concentration of compound in the plasma and buffer by HPLC -MS/MS.

Method io 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 bufl was isotonic buffered saline pH 7.4. Stock solutions of compound in dimethylsulpho; were prepared at a concentration of 0.5mM. Frozen pooled Human plasma was obtaii from volunteers.

I ₅

The stock DMSO solution of a compound was added to the plasma at a ratio of 10 μl c DMSO to each ml of plasma. This gave a 1% DMSO in plasma solution with each compound at a concentration of 5 μM.

₂ o 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. One prepared the cells were sealed and placed in an incubator box at 37°C. These cells we then rotated for a minimum of 4 hours to equilibrate.

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

0.013 μM, 0.05 μM, 0.25 μM and 1.25 μM which were injected in this order followec the buffer sample and then the plasma sample.

Calculation

The concentration of compound in the samples were determined using MassLynx veπ 4.1 software (produced by Waters/Micromass) that automatically calculated a calibrat 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 humar plasma (% bound) using the following equation;

buffer peak area . buffer injection volume

% bound = 100- 100

J plasma peak area plasma injection volume

Table 3 shows the measured human plasma protein binding figure using the procedure described above for some representative Examples.

Table 3

Methacholine Induced Bronchoconstriction in vivo

Dunkin-Hartley guinea-pigs (300 - 60Og) were supplied by a designated breeding establishment. Animals were dosed with test compound or vehicle either by inhalation

anaesthesia (5% halothane). Animals were allowed to recover from the anaesthesia pi to the measurement of bronchoconstriction. Up to 48 hours post-dosing guinea-pigs w terminally anaesthetized with sodium pentobarbitone (60 mg/kg), the trachea cannula for artificial ventilation and the jugular vein was cannulated for intravenous administr 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 vo of 5 ml/kg during surgical preparation. Lung function (lung resistance and complianc was measured in anaesthetised and ventilated guinea-pigs using a pulmonary measure Flexivent system (SCIREQ, Montreal, Canada) connected to the tracheal cannulae. T. animals were ventilated (quasi-sinusoidal ventilation pattern) at 60 breaths/min at a tic volume of 5 ml/kg. A positive end expiratory pressure of 2-3 CmH ₂ O 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 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 methachol as follows: f 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 (450-55Og) supplied by Harlan UK or David Hall, Staffs UK and acclima 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 H20, 1.5g/kj Once sufficient anaesthesia had developed (absence of toe pinch reflex) each animal h 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 i i: i : i *: ~f : * 4.1 1 1 _ _

s.c. pilocarpine administered under the skin at the back of the neck (0.6mg/kg @ 2ml/ The pad was removed, weighed and replaced with a new pre-weighed pad every 5 mil up to 15 minutes.

Saliva production was calculated by subtracting the pre-weighed weight of the pad frc each 5 minute period post weighed pad and these numbers added together to produce , 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 saliv production over 15 minutes.

Inhibition of saliva produced by the compound could be calculated by using the folio ¹ equation: ( l-(Test-baseline)/(Veh-baseline))* 100.