WO2009139707A1

WO2009139707A1 - Pharmaceutical product comprising a muscarinic receptor antagonist and a second active ingredient

Info

Publication number: WO2009139707A1
Application number: PCT/SE2009/050524
Authority: WO
Inventors: Richard Bull; Rhonan Ford; Andrew Mather; Antonio Mete
Original assignee: Argenta Discovery Ltd; AstraZeneca AB
Current assignee: Argenta Discovery Ltd; AstraZeneca AB
Priority date: 2008-05-13
Filing date: 2009-05-12
Publication date: 2009-11-19
Anticipated expiration: 2010-11-13
Also published as: GB0808709D0

Abstract

The invention provides a pharmaceutical product, kit or composition comprising a first active ingredient which is a selected muscarinic receptor antagonist, and a second active ingredient which is selected from a phosphodiesterase inhibitor, a modulator of chemokine receptor function, an inhibitor of kinase function, a protease inhibitor, a steroidal glucocorticoid receptor agonist, a non-steroidal glucocorticoid receptor agonist and a purinoceptor antagonist, of use in the treatment of respiratory diseases such as chronic obstructive pulmonary disease and asthma.

Description

PHARMACEUTICAL PRODUCT COMPRISING A MUSCARINIC RECEPTOR ANTAGONIST AND A SECOND ACTIVE INGREDIENT

The present invention relates to combinations of pharmaceutically active substances for use in the treatment of respiratory diseases, especially chronic obstructive pulmonary disease (COPD) and asthma.

The essential function of the lungs requires a fragile structure with enormous exposure to the environment, including pollutants, microbes, allergens, and carcinogens. Host factors, resulting from interactions of lifestyle choices and genetic composition, influence the response to this exposure. Damage or infection to the lungs can give rise to a wide range of diseases of the respiratory system (or respiratory diseases). A number of these diseases are of great public health importance. Respiratory diseases include Acute Lung Injury, Acute Respiratory Distress Syndrome (ARDS), occupational lung disease, lung cancer, tuberculosis, fibrosis, pneumoconiosis, pneumonia, emphysema, Chronic Obstructive Pulmonary Disease (COPD) and asthma.

Among the most common of the respiratory diseases is asthma. Asthma is generally defined as an inflammatory disorder of the airways with clinical symptoms arising from intermittent airflow obstruction. It is characterised clinically by paroxysms of wheezing, dyspnea and cough. It is a chronic disabling disorder that appears to be increasing in prevalence and severity. It is estimated that 15% of children and 5% of adults in the population of developed countries suffer from asthma. Therapy should therefore be aimed at controlling symptoms so that normal life is possible and at the same time provide basis for treating the underlying inflammation.

COPD is a term which refers to a large group of lung diseases which can interfere with normal breathing. Current clinical guidelines define COPD as a disease state characterized by airflow limitation that is not fully reversible. The airflow limitation is usually both progressive and associated with an abnormal inflammatory response of the lungs to noxious particles and gases. The most important contributory source of such particles and gases, at least in the western world, is tobacco smoke. COPD patients have a variety of symptoms, including cough, shortness of breath, and excessive production of sputum; such symptoms arise from dysfunction of a number of cellular compartments, including neutrophils, macrophages, and epithelial cells. The two most important conditions covered by COPD are chronic bronchitis and emphysema.

Chronic bronchitis is a long-standing inflammation of the bronchi which causes increased production of mucous and other changes. The patients' symptoms are cough and expectoration of sputum. Chronic bronchitis can lead to more frequent and severe respiratory infections, narrowing and plugging of the bronchi, difficult breathing and disability.

Emphysema is a chronic lung disease which affects the alveoli and/or the ends of the smallest bronchi. The lung loses its elasticity and therefore these areas of the lungs become enlarged. These enlarged areas trap stale air and do not effectively exchange it with fresh air. This results in difficult breathing and may result in insufficient oxygen being delivered to the blood. The predominant symptom in patients with emphysema is shortness of breath.

Therapeutic agents used in the treatment of respiratory diseases include muscarinic antagonists. Muscarinic receptors are a G-protein coupled receptor (GPCR) family having five family members M₁, M₂, M₃, M₄ and M5. Of the five muscarinic subtypes, three (M₁, M₂ and M₃) are known to exert physiological effects on human lung tissue. Parasympathetic nerves are the main pathway for reflex bronchoconstriction in human airways and mediate airway tone by releasing acetylcholine onto muscarinic receptors. Airway tone is increased in patients with respiratory disorders such as asthma and chronic obstructive pulmonary disease (COPD), and for this reason muscarinic receptor antagonists have been developed for use in treating airway diseases. Muscarinic receptor antagonsists, often called anticholinergics in clinical practice, have gained widespread acceptance as a first-line therapy for individuals with COPD, and their use has been extensively reviewed in the literature (e.g. Lee et al, Current Opinion in Pharmacology 2001,1, 223-229).

Whilst treatment with a muscarinic antagonist can yield important benefits, the efficacy of these agents is often far from satisfactory. Moreover, in view of the complexity of respiratory diseases such as asthma and COPD, it is unlikely that any one mediator can satisfactorily treat the disease alone. Hence there is a pressing medical need for new therapies against respiratory diseases such as COPD and asthma, in particular for therapies with disease modifying potential.

The present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is a muscarinic antagonist selected from:

(i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane X;

(R)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridazin-3-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane X

(R)-3 - [ 1 -(3 -Fluoro-pheny^-cycloheptanecarbonyloxy]- 1 -(pyrazin-2-ylcarbamoylmethyl)- l-azonia-bicyclo[2.2.2]octane X; (i?)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(isoxazol-3-ylcarbamoylmethyl)- l-azonia-bicyclo[2.2.2]octane X;

(i?)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane X;

(i?)-l-[(5-Fluoro-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phenyl-cycloheptanecarbonyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(R)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-3-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane X; and

(R)- 1 -[(2-Methyl-pyridin-4-ylcarbamoyl)-methyl]-3-(l -phenyl-cycloheptanecarbonyloxy)- l-azonia-bicyclo[2.2.2]octane X; wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and a second active ingredient which is selected from i) a phosphodiesterase inhibitor, ii) a modulator of chemokine receptor function, iii) an inhibitor of kinase function, iv) a protease inhibitor, v) a steroidal glucocorticoid receptor agonist, vi) a non-steroidal glucocorticoid receptor agonist, and vii) a purinoceptor antagonist.

A beneficial therapeutic effect may be observed in the treatment of respiratory diseases if a muscarinic antagonist according to the present invention is used in combination with a second active ingredient as specified above. The beneficial effect may be observed when the two active substances are administered simultaneously (either in a single pharmaceutical preparation or via separate preparations), or sequentially or separately via separate pharmaceutical preparations.

The pharmaceutical product of the present invention may, for example, be a pharmaceutical composition comprising the first and second active ingredients in admixture. Alternatively, the pharmaceutical product may, for example, be a kit comprising a preparation of the first active ingredient and a preparation of the second active ingredient and, optionally, instructions for the simultaneous, sequential or separate administration of the preparations to a patient in need thereof.

The first active ingredient in the combination of the present invention is a muscarinic antagonist selected from:

(R)-3-(\ -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane X;

(R)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridazin-3-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane X;

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

(i?)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-3-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane X; and (R)- 1 -[(2-Methyl-pyridin-4-ylcarbamoyl)-methyl]-3-(l -phenyl-cycloheptanecarbonyloxy)- l-azonia-bicyclo[2.2.2]octane X; wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid.

The muscarinic antagonists of the invention are selected members of a novel class of compound described in co-pending application PCT/GB2007/004350 which display high potency to the M3 receptor. The names of the muscarinic antagonists are IUPAC names generated by the Beilstein Autonom 2000 naming package , as supplied by MDL Information Systems Inc., based on the structures depicted in the examples, and stereochemistry assigned according to the Cahn-Ingold-Prelog system. For example, the name(i?)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 - azonia-bicyclo[2.2.2]octane, was generated from the structure:

The muscarinic antagonists of the present invention comprise an anion X associated with the positive charge on the quaternary nitrogen atom. The anion X may be any pharmaceutically acceptable anion of a mono or polyvalent (e.g. bivalent) acid. In an embodiment of the invention X may be an anion of a mineral acid, for example chloride, bromide, iodide, sulfate, nitrate or phosphate; or an anion of a suitable organic acid, for example toluenesulfonate (tosylate), edisylate (ethane- 1 ,2-disulfonate), isethionate (2- hydroxyethylsulfonate),lactate, oleic, maleate ((Z)-3-carboxy-acrylate), succinate (3- carboxy-propionate), malate ((S)-3-carboxy -2-hydroxy-propionate), p- acetamidobenzoateacetate, maleate, fumarate, citrate, oxalate, succinate, tartrate, methanesulphonate, p-toluenesulphonate, benzenesulphonate, napadisylate (naphthalene- 1,5-disulphonate) (e.g. a heminapadisylate), 2,5-dichlorobenzenesulphonate, (xinafoate) 1- hydroxy-2-naphthoate or l-hydroxynaphthalene-2-sulphonate.

In an embodiment of the invention, the muscarinic receptor antagonist is in the form of a bromide or napadisylate salt.

In an embodiment of the invention, the muscarinic receptor antagonist is selected from

(i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane bromide;

(i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane chloride;

(i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octanel-hydroxy-naphthalene-2-sulfonate; (i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)-l -azonia- bicyclo [2.2.2] octane2,5 -dichlorobenzenesulfonate;

(i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane hemi-naphthalene- 1 ,5-disulfonate;

(i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridazin-3-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane bromide;

(R)-3 - [ 1 -(3 -Fluoro-pheny^-cycloheptanecarbonyloxy]- 1 -(pyrazin-2-ylcarbamoylmethyl)-

1 -azonia-bicyclo[2.2.2]octane bromide;

(i?)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(isoxazol-3-ylcarbamoylmethyl)-

1 -azonia-bicyclo[2.2.2]octane bromide; (i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane bromide;

(i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane chloride;

(i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane hemi-naphthalene- 1, 5 -disulfonate;

(i?)-l-[(5-Fluoro-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phenyl-cycloheptanecarbonyloxy)- l-azonia-bicyclo[2.2.2]octane chloride; (i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-3-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane chloride; and

(R)- 1 -[(2-Methyl-pyridin-4-ylcarbamoyl)-methyl]-3-(l -phenyl-cycloheptanecarbonyloxy)- 1 -azonia-bicyclo [2.2.2] octane chloride .

In an embodiment of the invention, the muscarinic receptor antagonist is in the form of a napadisylate salt. When the muscarinic antagonist is a napadisylate salt the cation/anion ratio may vary, and for example may be 1 : 1 or 2: 1 , or a value between 1 : 1 and 2:1.

In an embodiment of the invention the muscarinic antagonist is in the form of a napadisylate salt wherein the napadisylate salt cation/anion ratio is 2:1. i.e. a hemi- napadisylate. Examples of muscarinic antagonists according to this embodiment include: (R)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane hemi-naphthalene-l,5-disulfonate; and (i?)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane hemi-naphthalene- 1 ,5-disulfonate.

In an embodiment of the invention, the muscarinic receptor antagonist is in the form of a 2,5-dichlorobenzene sulphonate or l-hydroxynaphthalene-2-sulphonate salt. Examples of muscarinic antagonists according to this embodiment include:

(R)-3 -( 1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane l-hydroxy-naphthalene-2-sulfonate; and

(R)-3 -( 1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo [2.2.2] octane 2,5 -dichlorobenzenesulfonate.

In an embodiment of the invention, the muscarinic receptor antagonist is in the form of a bromide salt.

The second active ingredient of the present invention is selected from i) a phosphodiesterase inhibitor, ii) a modulator of chemokine receptor function, iii) an inhibitor of kinase function, iv) a protease inhibitor, v) a steroidal glucocorticoid receptor agonist, vi) a non-steroidal glucocorticoid receptor agonist, and vii) a purinoceptor antagonist.

In an embodiment of the invention the second active ingredient is a phosphodiesterase inhibitor. Examples of a phosphodiesterase inhibitor that may be used according to this embodiment include a PDE4 inhibitor such as an inhibitor of the isoform PDE4D, a PDE3 inhibitor and a PDE5 inhibitor. Examples include the compounds (Z)-3-(3,5-dichloro-4-pyridyl)-2-[4-(2-indanyloxy-5-methoxy-2-pyridyl]propenenitrile,

N-[9-amino-4-oxo-l-phenyl-3,4,6,7-tetrahydropyrrolo[3,2,l -jk][l, 4]benzodiazepin-3(R)- yl]pyridine-3-carboxamide (CI-1044),

3-(benzyloxy)-l-(4-fluorobenzyl)-N-[3-(methylsulphonyl)phenyl]-lH-indole-2- carboxamide, (lS-exo)-5-[3-(bicyclo[2.2.1]hept-2-yloxy)-4-methoxyphenyl]tetrahydro-2(lH)- pyrimidinone (Atizoram),

N-(3,5,dichloro-4-pyridinyl)-2-[l-(4-fluorobenzyl)-5-hydroxy-lH-indol-3-yl]-2- oxoacetamide (AWD- 12-281), β-[3-(cyclopentyloxy)-4-methoxyphenyl]-l,3-dihydro-l,3-dioxo-2H-isoindole-2- propanamide (CDC-801),

N-[9-methyl-4-oxo-l-phenyl-3,4,6,7-tetrahydropyrrolo[3,2,l -jk][l, 4]benzodiazepin-3(R)- yl]pyridine-4-carboxamide (CI-1018), cis-[4-cyano-4-(3-cyclopentyloxy-4-methoxyphenyl)cyclohexane- 1 -carboxylic acid

(Cilomilast), 8-amino-l,3-bis(cyclopropylmethyl)xanthine (Cipamfylline),

N-(2,5-dichloro-3-pyridinyl)-8-methoxy-5-quinolinecarboxamide (D-4418),

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

2-methyl- 1 -[2-(I -methylethyl)pyrazolo[ 1 ,5-a]pyridin-3-yl]- 1 -propanone (Ibudilast),

2-(2,4-dichlorophenylcarbonyl)-3-ureidobenzofuran-6-yl methanesulphonate (Lirimilast), (-)-(R)-5-(4-methoxy-3-propoxyphenyl)-5-methyloxazolidin-2-one (Mesopram),

(-)-cis-9-ethoxy-8-methoxy-2-methyl-l,2,3,4,4a,10b-hexahydro-6-(4- diisopropylaminocarbonylphenyl)-benzo[c][l,6]naphthyridine (Pumafentrine), 3-(cyclopropylmethoxy)-N-(3,5-dichloro-4-pyridyl)-4-(difluoromethoxy)benzamide (Roflumilast), the N-oxide of Roflumilast,

5,6-diethoxybenzo[b]thiophene-2-carboxylic acid (Tibenelast), 2,3,6,7-tetrahydro-2-(mesitylimino)-9,10-dimethoxy-3-methyl-4H-pyrimido[6,l- a]isoquinolin-4-one (trequinsin) and

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

In an embodiment of the invention the second active ingredient is a modulator of chemokine receptor function. Examples of a modulator of chemokine receptor function that may be used in this embodiment include a CCR3 receptor antagonist, a CCR4 receptor antagonist, a CCR5 receptor antagonist and a CCR8 receptor antagonist.

In an embodiment of the invention the second active ingredient is a CCRl receptor antagonist.

In an embodiment of the invention, the second active ingredient is a CCRl receptor antagonist selected from: N- {2-[((25)-3- {[ 1 -(4-chlorobenzyl)piperidin-4-yl] amino} -2-hydroxy-2-methylpropyl)oxy]-

4-hydroxyphenyl} acetamide;

N- {5-chloro-2-[((25)-3- {[ 1 -(4-chlorobenzyl)piperidin-4-yl] amino } -2-hydroxy-2- methylpropyl)oxy]-4-hydroxyphenyl}acetamide;

2-{2-chloro-5-{[(2S)-3-(5-chloro-rH,3H-spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2- hydroxypropyl]oxy} -4-[(methylamino)carbonyl]phenoxy} -2-methylpropanoic acid; or pharmaceutically acceptable salts thereof.

In another embodiment of the present invention, the second active ingredient is a salt of N- {2-[((25)-3- {[ 1 -(4-chlorobenzyl)piperidin-4-yl] amino} -2-hydroxy-2-methylpropyl)oxy]-4- hydroxyphenyl} acetamide or TV- {5-Chloro-2-[((25)-3- {[ 1 -(4-chlorobenzyl)piperidin-4- yljamino} -2-hydroxy-2-methylpropyl)oxy]-4-hydroxyphenyl} acetamide, for example hydrochloride, hydrobromide, phosphate, sulfphate, acetate, ascorbate, benzoate, fumarate, hemifumarate, furoate, succinate, maleate, tartrate, citrate, oxalate, xinafoate, methanesulphonate or/?-toluenesulphonate salt.

In another embodiment of the present invention, the second active ingredient is a benzoate, furoate or hemifumarate salt of Λ/-{2-[((25)-3-{[l-(4-chlorobenzyl)piperidin-4-yl]amino}- 2-hydroxy-2-methylpropyl)oxy]-4-hydroxyphenyl}acetamide, as described in PCT/SE2006/000920, PCT/SE2006/000921 and PCT/SE2006/000922 (WO2007/015666, WO2007/015667 and WO2007/015668).

In another embodiment of the present invention, the second active ingredient is the hemifumarate, furoate, benzoate, 2-fluorobenzoate or 2,6-difiuorobenzoate salt of N- {5- Chloro-2-[((25)-3-{[l-(4-chlorobenzyl)piperidin-4-yl]amino}-2-hydroxy-2- methylpropyl)oxy]-4-hydroxyphenyl}acetamide.

In an embodiment of the present invention the second active ingredient is 2-{2-chloro-5- {[(2S)-3-(5-chloro-lΗ,3H-spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2- hydroxypropyl]oxy}-4-[(methylamino)carbonyl]phenoxy}-2-methylpropanoic acid or a pharmaceutically acceptable salt thereof. 2-{2-Chloro-5-{[(2S)-3-(5-chloro-l'H,3H- spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-4-

[(methylamino)carbonyl]phenoxy}-2-methylpropanoic acid may be prepared by methods according or analogous to those described in PCT/SE2007/000694 (WO2008/010765).

In an embodiment of the present invention the second active ingredient is 7V-{5-chloro-2- [((25)-3- {[ 1 -(4-chlorobenzyl)piperidin-4-yl] amino} -2-hydroxy-2-methylpropyl)oxy]-4- hydroxyphenyl}acetamide or a pharmaceutically acceptable salt thereof. 7V-{5-chloro-2- [((25)-3- {[ 1 -(4-chlorobenzyl)piperidin-4-yl] amino} -2-hydroxy-2-methylpropyl)oxy]-4- hydroxyphenyl}acetamide may be prepared by methods according or analogous to those described in WO2007/015664.

In an embodiment of the invention, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is N-{2-[((2S)-3-{[\-(4- chlorobenzyl)piperidin-4-yl]amino}-2-hydroxy-2-methylpropyl)oxy]-4- hydroxyphenyl} acetamide or a pharmaceutically acceptable salt thereof (e.g. benzoate, hemifumarate or furoate). In one aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane hemi-naphthalene-l,5-disulfonate. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane chloride. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane 2,5-dichlorobenzenesulfonate. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane 1 -hydroxy-naphthalene-2-sulfonate.

In an embodiment of the invention, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)-l-(pyridin-2-ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is Λ/-{2-[((25)-3-{[l-(4-chlorobenzyl)piperidin-4- yljamino} -2-hydroxy-2-methylpropyl)oxy]-4-hydroxyphenyl} acetamide or a pharmaceutically acceptable salt thereof (e.g. benzoate, hemifumarate or furoate). In one aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)-l-(pyridin-2-ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide. In another aspect of this embodiment, the muscarinic receptor antagonist is (R)- 3 -( 1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane chloride. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane hemi-naphthalene-l,5-disulfonate. In an embodiment of the invention, the muscarinic receptor antagonist is (i?)-l-[(5-Fluoro- pyridin-2-ylcarbamoyl)-methyl] -3 -( 1 -phenyl-cycloheptanecarbonyloxy)- 1 -azonia- bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is N-{2-[((2S)-3-{[\-(4- chlorobenzyl)piperidin-4-yl]amino}-2-hydroxy-2-methylpropyl)oxy]-4- hydroxyphenyl}acetamide or a pharmaceutically acceptable salt thereof (e.g. benzoate, hemifumarate or furoate). In one aspect of this embodiment, the muscarinic receptor antagonist is (i?)-l-[(5-Fluoro-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)- 1 -azonia-bicyclo[2.2.2]octane chloride.

In an embodiment of the invention, the muscarinic receptor antagonist is (i?)-3-[l-(3- Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(isoxazol-3-ylcarbamoylmethyl)-l-azonia- bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is N-{2-[((2S)-3-{[l-(4- chlorobenzyl)piperidin-4-yl]amino}-2-hydroxy-2-methylpropyl)oxy]-4- hydroxyphenyl}acetamide or a pharmaceutically acceptable salt thereof (e.g. benzoate, hemifumarate or furoate). In one aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(isoxazol-3- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide.

In an embodiment of the invention, the muscarinic receptor antagonist is (i?)-3 -(I -Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is N-{5-chloro-2-[((25)-3-{[l- (4-chlorobenzyl)piperidin-4-yl]amino}-2-hydroxy-2-methylpropyl)oxy]-4- hydroxyphenyl}acetamide or a pharmaceutically acceptable salt thereof (e.g. benzoate, hemifumarate or furoate). In one aspect of this embodiment, the muscarinic receptor antagonist is (R)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3 -(I -Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane hemi-naphthalene-l,5-disulfonate. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane chloride. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane 2,5-dichlorobenzenesulfonate. (R)-3- ( 1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane 1 -hydroxy-naphthalene-2-sulfonate.

In an embodiment of the invention, the muscarinic receptor antagonist is (R)-3 -(I -Phenyl- cycloheptanecarbonyloxy)-! -(pyridin-2-ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is Λ/-{5-chloro-2-[((25)-3-{[l-(4-chlorobenzyl)piperidin- 4-yl]amino} -2-hydroxy-2-methylpropyl)oxy]-4-hydroxyphenyl} acetamide or a pharmaceutically acceptable salt thereof (e.g. benzoate, hemifumarate or furoate). In one aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3 -(I -Phenyl- cycloheptanecarbonyloxy)-! -(pyridin-2-ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide. In another aspect of this embodiment, the muscarinic receptor antagonist is (R)- 3 -( 1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane chloride. In another aspect of this embodiment, the muscarinic receptor antagonist is (R)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane hemi-naphthalene-l,5-disulfonate.

In an embodiment of the invention, the muscarinic receptor antagonist is (i?)-l-[(5-Fluoro- pyridin-2-ylcarbamoyl)-methyl]-3-(l -phenyl-cycloheptanecarbonyloxy)-! -azonia- bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is N-{5-chloro-2-[((25)-3-{[l- (4-chlorobenzyl)piperidin-4-yl]amino}-2-hydroxy-2-methylpropyl)oxy]-4- hydroxyphenyl} acetamide or a pharmaceutically acceptable salt thereof (e.g. benzoate, hemifumarate or furoate). In one aspect of this embodiment, the muscarinic receptor antagonist is (R)-I -[(5 -Fluoro-pyridin-2-ylcarbamoyl)-methyl] -3 -(I -phenyl- cycloheptanecarbonyloxy)- 1 -azonia-bicyclo[2.2.2]octane chloride. In an embodiment of the invention, the muscarinic receptor antagonist is (i?)-3-[l-(3- Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(isoxazol-3-ylcarbamoylmethyl)-l-azonia- bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is N-{5-c\ύoro-2-[((2S)-3-{[\- (4-chlorobenzyl)piperidin-4-yl]amino}-2-hydroxy-2-methylpropyl)oxy]-4- hydroxyphenyl}acetamide or a pharmaceutically acceptable salt thereof (e.g. benzoate, hemifumarate or furoate). In one aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(isoxazol-3- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide.

In an embodiment of the invention, the muscarinic receptor antagonist (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane X , wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is 2-{2-Chloro-5-{[(2S)-3-(5- chloro- 1 Η,3H-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy} -4- [(methylamino)carbonyl]phenoxy}-2-methylpropanoic acid or a pharmaceutically acceptable salt thereof. In one aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane hemi-naphthalene-l,5-disulfonate. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane chloride. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane 2,5-dichlorobenzenesulfonate. (R)-3- (1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane 1 -hydroxy-naphthalene-2-sulfonate. In an embodiment of the invention, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)-l-(pyridin-2-ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is 2-{2-Chloro-5-{[(2S)-3-(5-chloro-l'H,3H-spiro[l- benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy} -4-

[(methylamino)carbonyl]phenoxy}-2-methylpropanoic acid or a pharmaceutically acceptable salt thereof. In one aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-2-ylcarbamoylmethyl)- l-azonia-bicyclo[2.2.2]octane bromide. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)-l-(pyridin-2-ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane hemi-naphthalene-l,5-disulfonate.

In an embodiment of the invention, the muscarinic receptor antagonist is (i?)-l-[(5-Fluoro- pyridin-2-ylcarbamoyl)-methyl] -3 -( 1 -phenyl-cycloheptanecarbonyloxy)- 1 -azonia- bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is 2-{2-Chloro-5-{[(2S)-3-(5- chloro- 1 Η,3H-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy} -4- [(methylamino)carbonyl]phenoxy}-2-methylpropanoic acid or a pharmaceutically acceptable salt thereof. In one aspect of this embodiment, the muscarinic receptor antagonist is (R)-I -[(5 -Fluoro-pyridin-2-ylcarbamoyl)-methyl] -3 -(I -phenyl- cycloheptanecarbonyloxy)- 1 -azonia-bicyclo[2.2.2]octane chloride.

In an embodiment of the invention, the muscarinic receptor antagonist is (i?)-3-[l-(3- Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(isoxazol-3-ylcarbamoylmethyl)-l-azonia- bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is 2-{2-Chloro-5-{[(2S)-3-(5- chloro- 1 Η,3H-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy} -4- [(methylamino)carbonyl]phenoxy}-2-methylpropanoic acid or a pharmaceutically acceptable salt thereof. In one aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(isoxazol-3- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide.

In an embodiment of the invention the second active ingredient is an inhibitor of kinase function. Examples of an inhibitor of kinase function that may be used in this embodiment include a p38 kinase inhibitor and an IKK inhibitor.

In an embodiment of the invention the second active ingredient is a protease inhibitor. Examples of a protease inhibitor that may be used in this embodiment include an inhibitor of neutrophil elastase or an inhibitor of MMP 12.

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

In an embodiment of the invention, the muscarinic receptor antagonist (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane X , wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is budesonide In one aspect of this embodiment, the muscarinic receptor antagonist (i?)-3 -(I -Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane bromide. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane hemi-naphthalene-l,5-disulfonate. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane chloride. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane 2,5-dichlorobenzenesulfonate. (R)-3- (1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane 1 -hydroxy-naphthalene-2-sulfonate.

In an embodiment of the invention, the muscarinic receptor antagonist is (R)-3 -(I -Phenyl- cycloheptanecarbonyloxy)-! -(pyridin-2-ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is budesonide. In one aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide. In another aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)-! -(pyridin-2-ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride. In another aspect of this embodiment, the muscarinic receptor antagonist is (R)- 3 -( 1 -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane hemi-naphthalene- 1 ,5-disulfonate.

In an embodiment of the invention, the muscarinic receptor antagonist is (i?)-l-[(5-Fluoro- pyridin-2-ylcarbamoyl)-methyl] -3 -( 1 -phenyl-cycloheptanecarbonyloxy)- 1 -azonia- bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is budesonide. In one aspect of this embodiment, the muscarinic receptor antagonist is (i?)-l-[(5-Fluoro-pyridin-2- ylcarbamoyl)-methyl]-3-(l -phenyl-cycloheptanecarbonyloxy)- 1 -azonia- bicyclo[2.2.2]octane chloride. In an embodiment of the invention, the muscarinic receptor antagonist is (i?)-3-[l-(3- Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(isoxazol-3-ylcarbamoylmethyl)-l-azonia- bicyclo[2.2.2]octane X, wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and the second active ingredient is budesonide. In one aspect of this embodiment, the muscarinic receptor antagonist is (i?)-3-[l-(3-Fluoro-phenyl)- cycloheptanecarbonyloxy]-l-(isoxazol-3-ylcarbamoylmethyl)-l-azonia- bicyclo[2.2.2]octane bromide.

In an embodiment of the invention the second active ingredient is a non-steroidal glucocorticoid receptor agonist. Examples of a modulator of a non-steroidal glucocorticoid receptor agonist that may be used in this embodiment include selective non-steroidal glucocorticoid receptor agonists. Non-steroidal glucocorticoid receptor agonists are described for example in WO2006/046916 and US6323199.

In an embodiment of the invention the second active ingredient is a purinoceptor antagonist, for example a P2X₇ receptor antagonist. Examples of P2X₇ receptor antagonists are described in WO00/61569, WO01/44170, WO01/94338, WO03/041707, WO03/080579, WO04/106305, WO05/009968, WO06/025784 and WO06/059945.

The combination of the present invention may provide a beneficial therapeutic effect in the treatment of respiratory diseases. Examples of such possible effects include improvements in one or more of the following parameters: reducing inflammatory cell influx into the lung, mild and severe exacerbations, FEVi (forced expiratory volume in one second), vital capacity (VC), peak expiratory flow (PEF), symptom scores and Quality of Life.

The muscarinic antagonist (first active ingredient) and second active ingredient of the present invention may be administered simultaneously, sequentially or separately to treat respiratory diseases. By sequential it is meant that the active ingredients are administered, in any order, one immediately after the other. They may still have the desired effect if they are administered separately, but when administered in this manner they will generally be administered less than 4 hours apart, more conveniently less than two hours apart, more conveniently less than 30 minutes apart and most conveniently less than 10 minutes apart.

The active ingredients of the present invention may be administered by oral or parenteral (e.g. intravenous, subcutaneous, intramuscular or intraarticular) administration using conventional systemic dosage forms, such as tablets, capsules, pills, powders, aqueous or oily solutions or suspensions, emulsions and sterile injectable aqueous or oily solutions or suspensions. The active ingredients may also be administered topically (to the lung and/or airways) in the form of solutions, suspensions, aerosols and dry powder . These dosage forms will usually include one or more pharmaceutically acceptable ingredients which may be selected, for example, from adjuvants, carriers, binders, lubricants, diluents, stabilising agents, buffering agents, emulsifying agents, viscosity-regulating agents, surfactants, preservatives, flavourings and colorants. As will be understood by those skilled in the art, the most appropriate method of administering the active ingredients is dependent on a number of factors.

In one embodiment of the present invention the active ingredients are administered via separate pharmaceutical preparations. Therefore, in one aspect, the present invention provides a kit comprising a preparation of a first active ingredient which is a muscarinic antagonist according to the present invention, and a preparation of a second active ingredient, and optionally instructions for the simultaneous, sequential or separate administration of the preparations to a patient in need thereof.

In another embodiment the active ingredients may be administered via a single pharmaceutical composition. Therefore, the present invention further provides a pharmaceutical composition comprising, in admixture, a first active ingredient, which is a muscarinic antagonist according to the present invention, and a second active ingredient, as defined above.

The pharmaceutical compositions of the present invention may be prepared by mixing the muscarinic antagonist (first active ingredient) with the second active ingredient and a pharmaceutically acceptable adjuvant, diluent or carrier. Therefore, in a further aspect of the present invention there is provided a process for the preparation of a pharmaceutical composition, which comprises mixing a muscarinic antagonist according to the present invention with a second active ingredient according to the present invention and a pharmaceutically acceptable adjuvant, diluent or carrier.

It will be understood that the therapeutic dose of each active ingredient administered in accordance with the present invention will vary depending upon the particular active ingredient employed, the mode by which the active ingredient is to be administered, and the condition or disorder to be treated.

In one embodiment of the present invention, the muscarinic antagonist (first active ingredient) according to the present invention is administered via inhalation. When administered via inhalation the dose of the muscarinic antagonist according to the present invention will generally be in the range of from 0.1 microgram (μg) to 5000 μg, 0.1 to 1000 μg, 0.1 to 500 μg, 0.1 to 100 μg, 0.1 to 50 μg, 0.1 to 5 μg, 5 to 5000 μg, 5 to 1000 μg, 5 to 500 μg, 5 to 100 μg, 5 to 50 μg, 5 to 10 μg, 10 to 5000 μg, 10 to 1000 μg, 10 to 500 μg, 10 to 100 μg, 10 to 50 μg, 20 to 5000 μg, 20 to 1000 μg, 20 to 500 μg, 20 to 100 μg, 20 to 50 μg, 50 to 5000 μg, 50 to 1000 μg, 50 to 500 μg, 50 to 100 μg, 100 to 5000 μg, 100 to 1000 μg or 100 to 500 μg. The dose will generally be administered from 1 to 4 times a day, conveniently once or twice a day, and most conveniently once a day.

In one embodiment of the present invention the second active ingredient of the present invention may conveniently be administered by inhalation. When administered via inhalation the dose of the second active ingredient will generally be in the range of from 0.1 to 50 μg, 0.1 to 40 μg, 0.1 to 30 μg, 0.1 to 20 μg, 0.1 to 10 μg, 5 to 10 μg, 5 to 50 μg, 5 to 40 μg, 5 to 30 μg, 5 to 20 μg, 5 to 10 μg, 10 to 50 μg, 10 to 40 μg 10 to 30 μg, or 10 to 20 μg. The dose will generally be administered from 1 to 4 times a day, conveniently once or twice a day, and most conveniently once a day.

In another embodiment of the present invention, the second active ingredient is administered orally. Oral administration of the second active ingredient may for example be used in a pharmaceutical product or kit wherein the other active ingredient(s) are administered by inhalation. When administered orally, satisfactory results will generally be obtained when the dose of the second active ingedient is in the range of from 5 to 1000 milligram (mg), 5 to 800mg, 5 to 600mg, 5 to 500mg, 5 to 400mg, 5 to 300mg, 5 to 200mg, 5 to lOOmg, 5 to 50mg, 20 to 1000 mg, 20 to 800mg, 20 to 600mg, 20 to 500mg, 20 to 400mg, 20 to 300mg, 20 to 200mg, 20 to lOOmg, 20 to 50mg, 50 to 1000 mg, 50 to 800mg, 50 to 600mg, 50 to 500mg, 50 to 400mg, 50 to 300mg, 50 to 200mg, 50 to lOOmg, 100 to 1000 mg, 100 to 800mg, 100 to 600mg, 100 to 500mg, 100 to 400mg, 100 to 300mg, or 100 to 200mg. The dose will generally be administered from 1 to 4 times a day, conveniently once or twice a day, and most conveniently once a day.

In one embodiment, the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is a muscarinic antagonist, and a second active ingredient, as defined herein above, wherein each active ingredient is formulated for inhaled administration.

In another embodiment of the present invention, the first active ingredient, which is a muscarinic antagonist, may be formulated for oral administration and the second active ingredient(s) ,as defined herein above, may be formulated for inhaled administration.

In yet another embodiment of the present invention, the first active ingredient, which is a muscarinic antagonist, may be formulated for inhalaed administration and the second active ingredient(s), as defined herein above, may be formulated for oral administration.

In yet a further embodiment of the present invention, the first active ingredient, which is a muscarinic antagonist, and the second active ingredient(s), as defined herein above, wherein each active ingredient is formulated for oral administration.

In an embodiment the pharmaceutical preparations of active ingredients may be administered simultaneously. In an embodiment the different pharmaceutical preparations of active ingredients may be administered sequentially.

In an embodiment the different pharmaceutical preparations of active ingredients may be administered separately.

The active ingredients of the present invention are conveniently administered via inhalation (e.g. topically to the lung and/or airways) in the form of solutions, suspensions, aerosols and dry powder formulations. For example metered dose inhaler devices may be used to administer the active ingredients, dispersed in a suitable propellant and with or without additional excipients such as ethanol, surfactants, lubricants or stabilising agents. Suitable propellants include hydrocarbon, chlorofluorocarbon and hydrofluoroalkane (e.g. heptafluoroalkane) propellants, or mixtures of any such propellants. Preferred propellants are P 134a and P227, each of which may be used alone or in combination with other propellants and/or surfactant and/or other excipients. Nebulised aqueous suspensions or, preferably, solutions may also be employed, with or without a suitable pH and/or tonicity adjustment, either as a unit-dose or multi-dose.

Dry powders and pressurized HFA aerosols of the active ingredients may be administered by oral or nasal inhalation. For inhalation, the compound is desirably finely divided. The finely divided compound preferably has a mass median diameter of less than 10 μm, and may be suspended in a propellant mixture with the assistance of a dispersant, such as a Cs- C₂₀ fatty acid or salt thereof, (for example, oleic acid), a bile salt, a phospholipid, an alkyl saccharide, a perfluorinated or polyethoxylated surfactant, or other pharmaceutically acceptable dispersant.

One possibility is to mix the finely divided compound of the invention with a carrier substance, for example, a mono-, di- or polysaccharide, a sugar alcohol, or another polyol. Suitable carriers are sugars, for example, lactose, glucose, raffinose, melezitose, lactitol, maltitol, trehalose, sucrose, mannitol; and starch. Alternatively the finely divided compound may be coated by another substance. The powder mixture may also be dispensed into hard gelatine capsules, each containing the desired dose of the active compound.

Another possibility is to process the finely divided powder into spheres which break up during the inhalation procedure. This spheronized powder may be filled into the drug reservoir of a multidose inhaler, for example, that known as the Turbuhaler^® in which a dosing unit meters the desired dose which is then inhaled by the patient. With this system the active ingredient, with or without a carrier substance, is delivered to the patient.

The combination of the present invention is useful in the treatment or prevention of respiratory-tract disorders such as chronic obstructive pulmonary disease (COPD), chronic bronchitis of all types (including dyspnoea associated therewith), asthma (allergic and non- allergic; 'wheezy-infant syndrome'), adult/acute respiratory distress syndrome (ARDS), chronic respiratory obstruction, bronchial hyperactivity, pulmonary fibrosis, pulmonary emphysema, and allergic rhinitis, exacerbation of airway hyperreactivity consequent to other drug therapy, particularly other inhaled drug therapy or pneumoconiosis (for example aluminosis, anthracosis, asbestosis, chalicosis, ptilosis, siderosis, silicosis, tabacosis and byssinosis).

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

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

The present invention further provides a pharmaceutical product, kit or pharmaceutical composition according to the invention for simultaneous, sequential or separate use in therapy.

The present invention further provides the use of a pharmaceutical product, kit or pharmaceutical composition according to the invention in the manufacture of a medicament for the treatment of a respiratory disease, in particular chronic obstructive pulmonary disease or asthma.

The present invention further provides a pharmaceutical product, kit or pharmaceutical composition according to the invention for use in the treatment of a respiratory disease, in particular chronic obstructive pulmonary disease or asthma.

The present invention still further provides a method of treating a respiratory disease which comprises simultaneously, sequentially or separately administering: (a) a (therapeutically effective) dose of a first active ingredient which is a muscarinic antagonist according to the present invention; and

(b) a (therapeutically effective) dose of a second active according to the present invention; to a patient in need thereof.

In the context of the present specification, the term "therapy" also includes "prophylaxis" unless there are specific indications to the contrary. The terms "therapeutic" and "therapeutically" should be construed accordingly. Prophylaxis is expected to be particularly relevant to the treatment of persons who have suffered a previous episode of, or are otherwise considered to be at increased risk of, the condition or disorder in question. Persons at risk of developing a particular condition or disorder generally include those having a family history of the condition or disorder, or those who have been identified by genetic testing or screening to be particularly susceptible to developing the condition or disorder.

The term "disease, unless stated otherwise, has the same meaning as the terms "condition" and "disorder" and are used interchangeably throughout the description and claims. The term "agent" and " active ingredient" means the compounds comprised in the combination of the present invention, e.g. a muscarine antagonist or a CCRl antagonist.

The pharmaceutical product, kit or composition of the present invention may optionally comprise a third active ingredient which third active ingredient is a substance suitable for use in the treatment of respiratory diseases. Examples of third active ingredients that may be incorporated into the present invention include those listed herein above as second active ingredients (i.e. a phosphodiesterase inhibitor, a modulator of chemokine receptor function, an inhibitor of kinase function, a protease inhibitor, a steroidal glucocorticoid receptor agonist, a non-steroidal glucocorticoid receptor agonist or a purinoceptor antagonist) it being recognised that they may be utilised as third active ingredients in embodiments where they have not been utilised as the second active ingredient .

In one embodiment of the invention, the third active ingredient is a β₂-adrenoceptor agonist. The β₂-adrenoceptor agonist may be any compound or substance capable of stimulating the β₂ -receptors and acting as a bronchodilator. Examples of β₂-adrenoceptor agonists that may be employed in the present invention include formoterol. The chemical name for formoterol is N-[2-hydroxy-5-[(l)-l-hydroxy-2-[[(l)-2-(4-methoxyphenyl)-l- methylethyl]amino]ethyl]phenyl]-formamide. The preparation of formoterol is described, for example, in WO 92/05147. In one aspect of this embodiment, the β₂-adrenoceptor agonist is formoterol fumarate. It will be understood that the invention encompasses the use of all optical isomers of formoterol and mixtures thereof including racemates. Thus for example, the term formoterol encompasses 7V-[2-hydroxy-5-[(lR)-l-hydroxy-2-[[(lR)-2- (4-methoxyphenyl)-l-methylethyl]amino]ethyl]phenyl]-formamide, 7V-[2-hydroxy-5-[(lS)- 1 -hydroxy-2-[[(l S)-2-(4-methoxyphenyl)- 1 -methylethyl]amino]ethyl]phenyl]-formamide and a mixture of such enantiomers, including a racemate.

In an alternative embodiment of the present invention, the pharmaceutical product, kit or pharmaceutical composition does not contain a β₂-adrenoceptor agonist.

The invention is illustrated by the following non-limiting Examples. In the Examples the following Figures are presented:

Figure 1 : X-ray powder diffraction pattern of muscarinic antagonist (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane bromide Crystalline Form A (Example 1). Figure 2: X-ray powder diffraction pattern of muscarinic antagonist (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane chloride Crystalline Form A (Example 2). Figure 3: X-ray powder diffraction pattern of muscarinic antagonist (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane chloride Crystalline Form A (Example 3). Figure 4: X-ray powder diffraction pattern of muscarinic antagonist (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane bromide Crystalline Form A (Example 4). Figure 5: X-ray powder diffraction pattern of muscarinic antagonist (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane l-hydroxy-naphthalene-2-sulfonate Crystalline Form A (Example 5).

Figure 6: X-ray powder diffraction pattern of muscarinic antagonist (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane 2,5-dichloro-benzenesulfonate Crystalline Form A (Example 6). Figure 7: X-ray powder diffraction pattern of muscarinic antagonist (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane hemi-naphthalene- 1 ,5-disulfonate Crystalline Form A

(Example 7).

Figure 8: X-ray powder diffraction pattern of muscarinic antagonist (i?)-3-(l-Phenyl- cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)- 1 -azonia- bicyclo[2.2.2]octane hemi-naphthalene- 1, 5 -disulfonate Crystalline Form A (Example 14)

Preparation of Muscarinic Antagonists

Muscarinic antagonists according to the present invention may be prepared as follows. Alternative salts to those described herein may be prepared by conventional chemistry using methods analogous to those described.

General Experimental Details for Preparation of Muscarinic Antagonists Unless otherwise stsated the following general conditions were used in the preparation of the Muscarinic Antagonists

All reactions were carried out under an atmosphere of nitrogen unless specified otherwise.

In the examples the NMR spectra were measured on a Varian Unity Inova spectrometer at a proton frequency of either 300 or 400 or 500 MHz, or on a Bruker DRX spectrometer at a proton frequency of 400 or 500 MHz, or on a Bruker Avance spectrometer with a proton frequency of 600 MHz or or on a Bruker Avance DPX 300 spectrometer with a proton frequency of 300 MHz. The MS spectra were measured on either an Agilent 1100 MSD G1946D spectrometer or a Hewlett Packard HPl 100 MSD G1946A spectrometer or a Waters Micromass ZQ2000 spectrometer. Names were generated using the Autonom 2000 (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.

X-Ray Powder Diffraction - XRPD - PANalytical CubiX PRO

Data was collected with a PANalytical CubiX PRO machine in θ - θ configuration over the scan range 2° to 40° 2 θ with 100-second exposure per 0.02° increment. The X-rays were generated by a copper long-fine focus tube operated at 45kV and 4OmA. The wavelength of the copper X-rays was 1.5418 A . The Data was collected on zero background holders on which ~ 2 mg of the compound was placed. The holder was made from a single crystal 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.

X-Ray Powder Diffraction -PANalytical X-Pert

Data was collected using a PANalytical X-Pert machine in 2Θ - # configuration over the scan range 2° to 40° 2 θ with 100-second exposure per 0.02° increment. The X-rays were generated by a copper long-fine focus tube operated at 45kV and 4OmA. The wavelengths of the copper X-rays was 1.5418 A . The Data was collected on zero background holders on which ~ 2 mg of the compound was placed. The holder was made from a single crystal 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.

Differential Scanning Calorimetry (DSC) thermograms were measured using a TA Instruments QlOOO DSC Differential Scanning Calorimeter, with aluminium pans and pierced lids. The sample weights varied between 0.5 to 5 mg. The procedure was carried out under a flow of nitrogen gas (50 mL/min) and the temperature studied from 25 to 300⁰C at a constant rate of temperature increase of 10⁰C per minute.

Thermogravimetric Analysis (TGA) thermograms were measured using a TA Instruments Q500 TGA Thermogravimetric Analyser, with 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 from 25 up to 200-300⁰C at a constant rate of temperature increase of 10⁰C per minute.

Gravimetric Vapour Sorption (GVS) profiles were measured using a Surface Measurements Systems Dynamic Vapour Sorption DVS-I, or DVS Advantage GVS instruments. The solid sample ca. 1-5 mg was placed into a glass or wire mesh vessel and the weight of the sample was 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

DSC = Differential Scanning Calorimeter GVS = Gravimetric vapour sorption

TGA = Thermogravimetric analysis

XRPD = X-Ray Powder Diffraction

HATU = O-(7-Azabenzotriazol-l-yl)-Λ/,Λ/,N',N'-tetramethyluronium hexafluorophospahte MeCN - Acetonitrile

MeOH = methanol

RT = Room Temperature

Rt = retention time

THF = tetrahydrofuran Satd = saturated

Example 1: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2] octane bromide a) 1-Phenyl-cycloheptanol

To magnesium (1.2 g) in anhydrous tetrahydrofuran (60 mL) under an environment of nitrogen was added a crystal of iodine followed by bromobenzene (7.85 g) at such a rate 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 minutes saturated aqueous ammonium chloride (10 mL) was added and the reaction was partitioned between water (100 mL) and isohexane (100 mL). The organic layer was dried (MgSO₄) and evaporated to afford the sub-titled compound (7.6 g) as an oil.

¹H NMR (299.946 MHz, CDCl₃) δ 7.53 - 7.47 (m, 2H), 7.36 - 7.29 (m, 2H), 7.26 - 7.19 (m, 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 mL) and sodium hydride (60% in oil, 2.0 g) added. The reaction was stirred at 60⁰C for 5 minutes and iodomethane (7.1 g) added. The mixture was maintained at 60⁰C overnight and then further quantities of sodium hydride (60% in oil, 2.0 g) and iodomethane (7.1 g) were added and the reaction was refluxed for 70 hours. The reaction mixture was partitioned between water (100 mL) and z^'søhexane (100 mL) and the organic layer separated, dried (MgSO₄) and evaporated to afford the sub-titled compound (11.31 g).

¹H NMR (300 MHz, CDCl₃) δ 7.43 - 7.37 (m, 2H), 7.37 - 7.30 (m, 2H), 7.24 - 7.19 (m, 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 120⁰C in mineral oil under an environment of nitrogen for 30 minutes and then cooled to room temperature. The oil was removed and replaced with ether (100 mL) and 1-methoxy-l-phenyl-cycloheptane (Example Ib) (4.9 g) was added and the reaction was stirred under nitrogen overnight at room temperature. The reaction was cooled to -78°C and solid carbon dioxide (-20 g) was added with stirring. The reaction was allowed to warm to room temperature and water (150 mL) was added carefully under an environment of nitrogen. The aqueous layer was separated, neutralised with concentrated hydrochloric acid and extracted with diethyl ether (150 mL). The organic layer was dried (MgSO₄) and evaporated afford to the sub-titled compound (4.15 g) as an oil.

¹H NMR (300 MHz, CDCl₃) δ 7.40 - 7.20 (m, 5H), 2.49 - 2.35 (m, 2H), 2.16 - 2.03 (m, 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 methanol (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 water (100 mL), saturated sodium bicarbonate (50 mL) and water (100 mL), dried (MgSO₄) and evaporated to afford the sub-titled compound (3.5 g) as an oil.

¹H NMR (300 MHz, CDCl₃) 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 (i?)-(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) in a Dean and Stark apparatus for 24 hours. Heptane (20 mL) was replaced with toluene (20 mL) and the reflux was continued for 3 days. The reaction was partitioned between water (50 mL) and ether (50 mL) and the ether layer was separated, dried (MgSO₄) and evaporated. The crude product was purified by column chromatography on silica eluting with ethyl acetate / triethylamine (99/1) to afford the titled compound as an oil (0.83 g).

m/e 328 [M+H]⁺

¹H NMR (300 MHz, CDCl₃) δ 7.35 - 7.27 (m, 4H), 7.23 - 7.16 (m, IH), 4.78 - 4.71 (m, IH), 3.12 (ddd, IH), 2.79 - 2.32 (m, 7H), 2.16 - 1.98 (m, 2H), 1.91 - 1.80 (m, IH), 1.70 - 1.34 (m, 12H).

f) 2-Bromo-7V-pyrazin-2-yl-acetamide

To a stirred suspension of pyrazin-2-ylamine (1.878 g) and potassium carbonate (8.19 g) in dichloromethane (25 mL) was added 2-bromoacetyl bromide (1.72 mL). The reaction was stirred overnight and then washed with water (2 x 50 mL). The organic layer was dried (MgSO₄) and concentrated to afford the sub-titled compound as a solid (0.700 g).

¹H NMR (400 MHz, CDCl₃) δ 9.51 (d, IH), 8.63 (s, IH), 8.42 (d, IH), 8.30 (dd, IH), 4.06 (s, 2H).

Example 1: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide Crystalline Form A

1 -Phenyl-cycloheptanecarboxylic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ie) (0.200 g) and 2-bromo-7V-pyrazin-2-yl-acetamide (Example If) (0.132 g) were dissolved in acetonitrile (1 mL) and left to stand overnight. The resulting solid was filtered and washed with acetonitrile (2 x 1 mL), and diethyl ether (3 mL). The dried solid was recrystallised from acetone (15 mL) and diethyl ether (10 mL) to afford the titled compound (0.240 g).

m/e 463 [M]⁺

¹H NMR (400 MHz, DMSO-D₆) δ 11.37 (s, IH), 9.28 (s, IH), 8.50 - 8.46 (m, 2H), 7.39 - 7.30 (m, 4H), 7.27 - 7.21 (m, IH), 5.16 - 5.08 (m, IH), 4.33 (s, 2H), 4.17 - 4.07 (m, IH), 3.69 - 3.56 (m, 4H), 3.48 - 3.38 (m, IH), 2.44 - 2.26 (m, 3H), 2.25 - 2.04 (m, 2H), 2.03 - 1.87 (m, 3H), 1.85 - 1.71 (m, IH), 1.68 - 1.45 (m, 8H).

Analysis of Example 1: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide Crystalline Form A

A sample of crystalline Example 1 Crystalline Form A obtained by the procedure described above was analysed by XRPD (PANalytical X'Pert or CubiX system), DSC and TGA.

The melting temperature of Example 1 bromide Form A as determined by DSC was found to be 202⁰C (onset) (±2°C). Weight loss observed prior to melting by TGA was 2.7%. GVS determination gave a 3% weight increase (%w/w) at 80% RH (±0.2%).

An XRPD spectrum of Example 1 bromide Form A is presented in Figure 1. Example 2: (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2] octane chloride

a) 2-Chloro-7V-pyrazin-2-yl-acetamide

To a stirred suspension of pyrazin-2-ylamine (4.6 g) and potassium carbonate (20.05 g) in dichloromethane (50 mL) was added 2-chloroacetyl chloride (3.85 mL). The reaction was stirred overnight and then washed with water (2 x 50 mL). The organic layer was dried (MgSO₄) and concentrated to give a solid which was purifed by column chromatography on Silica eluting with ethyl acetate / z^'søhexane (5:95) to afford the sub-titled compound as a white solid (2.2 g).

m/e 172 [M+H]⁺ 1H NMR (400 MHz, DMSO-D₆) δ 11.12 (s, IH), 9.31 (d, IH), 8.44 (dd, IH), 8.41 (d, IH), 4.40 (s, 2H).

Example 2: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride Crystalline Form A

1-Phenyl-cycloheptanecarboxylic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ie) (0.55 g) and 2-chloro-7V-pyrazin-2-yl-acetamide (Example 2a) (0.288 g) were stirred in acetonitrile (4 mL) overnight. Further acetonitrile (14 mL) was added and the mixture stirred for 2 hours. The solid was collected by filtration and washed with diethyl ether (4 x 10 mL) to afford the titled compound as a solid (0.735 g).

m/e 463 [M]⁺ 1H NMR (400 MHz, DMSO-D₆) δ 11.52 (s, IH), 9.28 (s, IH), 8.49 - 8.45 (m, 2H), 7.38 - 7.31 (m, 4H), 7.26 - 7.21 (m, IH), 5.15 - 5.10 (m, IH), 4.44 (d, IH), 4.40 (d, IH), 4.14 (ddd, IH), 3.73 - 3.59 (m, 4H), 3.48 - 3.38 (m, IH), 2.42 - 2.29 (m, 2H), 2.23 - 2.12 (m, 2H), 2.04 - 1.87 (m, IH), 1.83 - 1.73 (m, 3H), 1.71 - 1.45 (m, 9H).

Analysis of Example 2: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride Crystalline Form A

A sample of crystalline Example 2 Crystalline Form A obtained by the procedure described above was analysed by XRPD (PANalytical X'Pert or CubiX system), DSC and TGA.

The melting temperature of Example 2 chloride Form A as determined by DSC was found to be 215°C (onset) (±2°C). GVS determination gave a 9% weight increase (%w/w) at 80% RH (±0.2%).

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

Example 3: (if)-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 dropwise to a stirred (overhead stirrer) solution of cycloheptanecarbonitrile (50 g) in 229 mL diethyl ether under nitrogen at such a rate as to maintain gentle reflux. The reaction mixture was then heated at reflux for 3 hours. TLC indicated no starting material present in the reaction mixture. The reaction mixture was allowed to cool to room temperature and stood under nitrogen overnight. The reaction mixture was cooled to 0⁰C and treated dropwise with 102 mL 4N HCl(aq) keeping the temperature below 20⁰C. 4N sulfuric acid (203 mL) was added dropwise rapidly to start with and then more quickly towards the end. The ice bath was removed and the diethyl ether was distilled off. The reaction mixture was heated at 80-90⁰C for 3.5 hours then allowed to cool to room temperature and stood overnight. The mixture was diluted with ether (approx 450 mL) and water (100 mL). The layers were separated and the aqueous layer was extracted with ether (2 x 400 mL). The organic layers were combined and washed with saturated aqueous sodium hydrogen carbonate (600 mL) and brine (600 mL), dried over magnesium sulphate, filtered and evaporated to give the sub-titled compound as an orange liquid (86.5 g).

¹H NMR (300 MHz, CDCl₃) 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-cycloheptyl)-phenyl-methanone

Sulfuryl chloride (210 mL) was added dropwise to neat cycloheptyl-phenyl-methanone (Example 3a) (86.5 g) at 0⁰C over approximately 1 hour. Gas evolution and an exotherm were observed. The internal temperature was kept below 15°C during the addition and the evolved gas was scrubbed by passing through a 10.2M aqueous solution of NaOH. The reaction mixture was heated to reflux overnight. TLC indicated no starting material remained. The reaction mixture was cooled to 0⁰C and poured slowly onto ice (1 L) with stirring. The layers were separated and the aqueous layer was extracted with ether (2 x 400 mL). The combined organic layers were washed with water (600 mL), saturated aqueous sodium hydrogen carbonate (600 mL), and brine (600 mL), dried over magnesium sulphate, filtered and evaporate to give the sub-titled compound as a brown oil (100 g).

¹H NMR (400 MHz, CDCl₃) δ 8.10-8.06 (d, 2H), 7.52-7.46 (t, IH), 7.44-7.36 (t, 2H), 2.50 (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 3b) (100 g) in 750 mL dioxane was treated dropwise rapidly with a cloudy solution of silver nitrate (137 g) in water (85 mL) causing a precipitate to form. The reaction mixture was heated to 75°C for 4.5 hours. TLC showed no starting material remaining. The reaction mixture was cooled to room temperature then filtered and concentrated to approximately 200 mL. Water (200 mL) and ether (300 mL) were added and the layers separated. The aqueous layer was extracted with ether (2 x 250 mL). The combined organic layers were extracted with 10% aqueous sodium carbonate (3 x 250 mL). The combined basic extracts were heated up to 90⁰C over 40 minutes and then cooled to room temperature and acidified with concentrated HCl (aq). The resulting brown solid was filtered off, washed with water (x2) and dried under vacuum at 50⁰C. Crystallisation from hot ethanol (40 mL) gave the sub-titled compound as pale brown crystals (9.83 g).

¹H NMR (400 MHz, CD₃OD) δ 7.36-7.26 (m, 4H), 7.21-7.15 (m, IH), 2.43-2.35 (m, 2H), 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 3c) (9.8 g) in methanol (85 mL) and toluene (300 mL) under an atmosphere of nitrogen. TLC after 45minutes showed no starting material present. The reaction mixture was concentrated under vacuum and the crude product was purified by column chromotography eluting with 0-10% ethyl acetate / cyclohexane. The relevant fractions were combined to give the product as a pale yellow oil (9.25 g).

¹H NMR (300 MHz, CD₃OD) δ 7.32-7.24 (m, 4H), 7.21-7.12 (m, IH), 3.60 (s, 3H), 2.43- 2.32 (m, 2H), 2.07-1.96 (m, 2H), 1.65-1.58 (m, 8H).

H A solution of (i?)-(3)-quinuclidinol (10.13 g) and 1-phenyl-cycloheptanecarboxylic acid methyl ester (Example 3d) (9.25 g) in toluene (90 mL) was heated to reflux with a Dean- Stark trap for 30 min. The reaction mixture was allowed to cool to room temperature and the trap was removed. Sodium hydride (60% dispersion in mineral oil) (3.19 g) was added portionwise under nitrogen and the reaction mixture was heated to reflux overnight under nitrogen. TLC showed no starting material remaining. 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 the crude product which was purified by silica gel chromatography eluting with EtOAc containing 1% triethylamine. The relevant fractions were combined and evaporated to give the sub-titled compound as a colourless oil (7.63 g).

¹H NMR (400 MHz, CD₃OD) δ 7.34-7.28 (m, 4H), 7.23-7.17 (m, IH), 4.80-4.75 (m, IH), 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 (m, IH), 1.69-1.54 (m, 10H), 1.54-1.42 (m, IH), 1.35-1.24 (m, IH).

f) 2-Chloro-7V-pyridin-2-yl-acetamide

A solution of 2-amino-pyridine (1.0 g) in dry dichlorome thane (10.6 mL) under nitrogen at O⁰C was treated with triethylamine (1.63 mL) followed by slow addition of chloroacetyl chloride (0.93 mL). The reaction mixture was allowed to warm up to room temperature. After 2 hours, the mixture was partitioned between dichloromethane and water. The phases were separated and the aqueous layer was extracted with dichloromethane (x2). The combined organic layer was washed with brine, dried over magnesium sulphate, filtered and concentrated to give the crude product which was purified by silica gel chromatography eluting with 0-30% ethyl acetate / cyclohexane. The relevant fractions were combined and evaporated to give the title compound (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 portion of the material from refluxing acetonitrile (2.4 mL) gave the sub-titled compound as a pink solid (0.73 g).

¹H NMR (400 MHz, CDCl₃): 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 3: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane chloride Crystalline Form A

H

A solution of 1-phenyl-cycloheptanecarboxylic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 3e) (254 mg) in acetonitrile (5 mL) was treated with 2-chloro-7V-pyridin-2- yl-acetamide (Example 3f) (46 mg) and the resulting yellow solution was 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, washed with ether and dried under vacuum to give the title compound (217 mg) as an off-white solid. Purification was achieved by crystallisation from refiuxing acetonitrile (20 mL) to give 98 mg of the title compound as a white crystalline solid.

m/e 462 [M]⁺

¹H NMR (400 MHz, DMSO-D₆): δ 11.09 (s, IH), 8.34-8.32 (d, IH), 7.97 (d, IH), 7.85- 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, 2H), 1.99-1.81 (m, 3H), 1.78-1.66 (m, IH), 1.77-1.19 (m, 9H).

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

A sample of crystalline Example 3 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 3 chloride Form A as determined by DSC was found to be 239°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% RH (±0.2%).

An XRPD spectrum of Example 3 chloride Form A is presented in Figure 3. Example 4: (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2] octane bromide

a) 2-Bromo-7V-pyridin-2-yl-acetamide

To a solution of 2-aminopyridine (48.8 mmol) in anhydrous THF (98 mL) at room temperature was added Et₃N (58.6 mmol) and bromoacetyl bromide (58.6 mmol) dropwise. The mixture was stirred overnight and quenched with sat. NaHCO_{3 (aq)}. EtOAc was added to the mixture and the layers separated. The aqueous phase was extracted with EtOAc and the combined organics dried (MgSO₄) and concentrated in vacuo to a brown solid. Purification by flash silica gel chromatography eluting with 1-2% MeOH / dichloromethane gave the sub-titled compounds as a yellow solid (1.14 g).

¹H NMR (400 MHz, CDCl₃): δ 8.75 (s, IH), 8.26 (ddd, IH), 8.10 (d, IH), 7.67 (ddd, IH), 7.03 (ddd, IH), 3.94 (s, 2H).

Example 4: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide Crystalline Form A

H

1 -Phenyl-cycloheptanecarboxylic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 3e) (0.79 mmol) and 2-bromo-7V-pyridin-2-yl-acetamide (Example 4a) (0.87 mmol) were 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 column chromatography eluting with 2-8% MeOH/dichloromethane to give a tan solid. The solid was dissolved up in refluxing MeCN and the solution was allowed to cool down to room temperature. The resulting crystals were filtered off and washed with a small quantity of cold MeCN to give the title compound (211 mg) as a white crystalline solid.

m/e 462 [M]⁺

¹H NMR (400 MHz, DMSO-D₆): δ 11.02 (s, IH), 8.33 (ddd, IH), 7.97 (d, IH), 7.86-7.80 (m, IH), 7.32-7.25 (m, 4H), 7.23-7.12 (m, 2H), 5.09-5.04 (m, IH), 4.23 (s, 2H), 4.06 (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, 2H), 1.98-1.83 (m, 3H), 1.78-1.66 (m, IH), 1.65-1.39 (m, 9H).

Analysis of Example 4: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide Crystalline Form A

A sample of crystalline Example 4 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 4 bromide Form A as determined by DSC was found to be 230⁰C (onset) (±2°C). Weight loss observed prior to melting by TGA was negligible. GVS determination gave a neglible weight increase (%w/w) at 80% RH (±0.2%).

An XRPD spectrum of Example 4 bromide Form A is presented in Figure 4.

Example 5: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane l-hydroxy-naphthalene-2- sulfonate Crystalline Form A

H

(i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane chloride (Example 2) (100 mg) and l-hydroxynaphthalene-2-sulfonic acid potassium salt (200 mg) were partitioned between water (10 mL) and dichloromethane (25 mL) in a separating funnel. The dichloromethane was separated and washed with water (10 mL) and the organic layer was dried, evaporated to a solid which was recrystallised from acetonitrile to afford the titled compound as a solid (97 mg).

m/e 463 [M]⁺

¹H NMR (400 MHz, DMSO-D₆) δ 11.61 (s, IH), 11.36 (s, IH), 9.28 (s, IH), 8.49 - 8.45 (m, 2H), 8.17 (d, IH), 7.81 (d, IH), 7.56 - 7.46 (m, 3H), 7.38 - 7.29 (m, 5H), 7.27 - 7.21 (m, IH), 5.16 - 5.09 (m, IH), 4.30 (s, 2H), 4.16 - 4.07 (m, IH), 3.68 - 3.54 (m, 4H), 3.48 - 3.35 (m, IH), 2.42 - 2.27 (m, 2H), 2.25 - 2.10 (m, 2H), 2.03 - 1.89 (m, 3H), 1.84 - 1.71 (m, IH), 1.66 - 1.51 (m, 9H).

Analysis of Example 5: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane l-hydroxy-naphthalene-2- sulfonate Crystalline Form A

A sample of crystalline Example 5 Crystalline Form A obtained by the procedure described above was analysed by XRPD (PANalytical X'Pert or CubiX system) and DSC.

The melting temperature of Example 5 l-hydroxy-naphthalene-2-sulfonate Form A as determined by DSC was found to be 193°C (onset) (±2°C). GVS determination gave a neglible weight increase, near 0.3% (%w/w) at 80% RH (±0.2%). An XRPD spectrum of Example 5 l-hydroxy-naphthalene-2-sulfonate Form A is presented in Figure 5.

Example 6: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyrj-l-azonia-bicycloβ^^] octane 2,5-dichloro-benzenesulfonate Crystalline Form A

(i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane chloride (Example 2) (100 mg) was suspended between water (10 mL) and dichloromethane (25 mL) in a separating funnel. An aqueous solution of 2,5- dichlorobenzenesulfonic acid sodium salt (0.1M, 8 mL) was added and the mixture shaken. The dichloromethane was separated and washed with water (10 mL) and the organic layer was dried, evaporated to a solid which was recrystallised from acetonitrile / diethyl ether to afford the titled compound (81 mg).

m/e 463 [M]⁺

¹H NMR (400 MHz, DMSO-D₆) δ 11.36 (s, IH), 9.27 (s, IH), 8.49 - 8.44 (m, 2H), 7.83 (d, IH), 7.43 - 7.31 (m, 6H), 7.27 - 7.22 (m, IH), 5.16 - 5.09 (m, IH), 4.36 - 4.25 (m, 2H), 4.16 - 4.07 (m, IH), 3.69 - 3.55 (m, 4H), 3.48 - 3.36 (m, IH), 2.42 - 2.28 (m, 2H), 2.23 - 2.10 (m, 2H), 2.03 - 1.87 (m, 3H), 1.83 - 1.72 (m, IH), 1.69 - 1.46 (m, 9H).

Analysis of Example 6: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2] octane 2,5-dichloro-benzenesulfonate Crystalline Form A A sample of crystalline Example 6 Crystalline Form A obtained by the procedure described above was analysed by XRPD (PANalytical X'Pert system) and DSC.

The melting temperature of Example 6 2,5-dichloro-benzenesulfonate Form A as determined by DSC was found to be 158°C (onset) (±2°C). GVS determination gave a neglible weight increase, near 0.2% (%w/w) at 80% RH (±0.2%).

An XRPD spectrum of Example 6 2,5-dichloro-benzenesulfonate Form A is presented in Figure 6.

Example 7: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyO-l-azonia-bicycloβ.l.l] octane hemi-naphthalene-l,5-disulfonate Crystalline Form A

(i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane chloride (Example 2) (100 mg) in dichloromethane (25 mL) was washed with 4 x 10 mL of an aq. solution of naphthalene- 1,5 -disulfonic acid di sodium salt (made by addition of 2.88 g acid to 1.68 g of sodium bicarbonate in 100 mL). The organic phase was collected and dried (MgSO₄) then concentrated to dryness. The residue was dissolved in acetone (1 mL) and diethyl ether (3 mL) and the solution allowed to crystallise to afford the titled compound (78 mg).

m/e 463 [M]⁺ ¹H NMR ^OO MHZ₅ DMSO-D₆) O 11.10 (S, IH), 9.22 (S, IH), 8.94 (d, IH), 8.43 (d, 2H), 7.94 (d, IH), 7.28 - 7.38 (m, 5H), 7.17 - 7.26 (m, IH), 5.09 - 5.17 (m, IH), 4.25 - 4.32 (m, 2H), 4.05 - 4.16 (m, IH), 3.54 - 3.68 (m, 4H), 3.35 - 3.50 (m, IH), 2.96 - 3.04 (m, 3H), 2.28 - 2.41 (m, IH), 2.10 - 2.26 (m, IH), 1.90 - 2.10 (m, 2H), 1.73 - 1.86 (m, IH), 1.48 - 1.72 (m, 9H).

Analysis of Example 7: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyrazin-2- ylcarbamoylmethyO-l-azonia-bicycloβ.l.l] octane hemi-naphthalene-l,5-disulfonate Crystalline Form A

A sample of crystalline Example 7 Crystalline Form A obtained by the procedure described above was analysed by XRPD (PANalytical X'Pert or CubiX system) and DSC.

The melting temperature of Example 7 hemi-naphthalene-l,5-disulfonate Form A as determined by DSC was found to be 222°C (onset) (±2°C). GVS determination gave a 1.6% weight increase (%w/w) at 80% RH (±0.2%).

An XRPD spectrum of Example 7 hemi-naphthalene-l,5-disulfonate Form A is presented in Figure 7.

Example 8: (if)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(pyrazin-2- ylcarbamoylmethyrj-l-azonia-bicycloβ.l.l] octane bromide

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

(3-Fluoro-phenyl)-acetic acid methyl ester (4.30 g) was dissolved in tetrahydrofuran (20 mL) and cooled to -78°C. Lithium όzs(trimethylsilyl)amide (25.6 mL, IM THF solution) was added and the solution was stirred for 30 minutes. 4-Bromo-but-l-ene (2.60 mL) was added and the reaction was allowed to warm to room temperature and stirred for an hour. The reaction was again cooled to -78°C. Lithium όzs(trimethylsilyl)amide (25.6 mL, IM THF solution) was added and the solution was stirred for 30 minutes. 4-Bromo-l-butene (2.60 mL) was added and the reaction was allowed to warm to room temperature and stirred for an hour. The reaction was again cooled to -78°C and further aliquots of Lithium όzs(trimethylsilyl)amide (25.6 mL, IM THF solution) and 4-bromo-l-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 ether (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 eluting with ethyl acetate / iso hexane (1 / 99) to afford the sub-titled compound (5.0 g).

m/e 277 [M+H]⁺

b) l-(3-Fluoro-phenyl)-cyclohept-4-enecarboxylic acid methyl ester

To 2-but-3-enyl-2-(3-fluoro-phenyl)-hex-5-enoic acid methyl ester (Example 8a) (5.0 g) in dichloromethane (100 mL) was added Grubbs Catalyst (2nd Generation, Sigma- Aldrich Company Ltd) (0.05 g). The mixture was warmed to reflux under nitrogen. After 20 hours the reaction was cooled to room temperature, evaporated to an oil and purified by column chromatography on silica eluting with ethyl acetate / z^'søhexane (5 / 95) to yield an oil. Analysis of the product showed that significant amounts of starting material was present in 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]⁺

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

l-(3-Fluoro-phenyl)-cyclohept-4-enecarboxylic acid methyl ester (Example 8b) (1.09 g) was disolved in methanol (20 mL), palladium on carbon (50 mg) added and mixture stirred under 4 atm of hydrogen overnight. The solution was filtered and evaporated to afford the sub-titled compound (1.09 g).

m/e 251 [M+H]⁺

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

H l-(3-Fluoro-phenyl)-cycloheptanecarboxylic acid methyl ester (Example 8c) (0.280 g) was dissolved in toluene (100 mL) and (i?)-quinuclidin-3-ol (0.320 g) was added. Toluene (10 mL) was distilled off in a Dean and Stark apparatus and after cooling sodium hydride (10 mg) was added. The reaction was refluxed in a Dean and Stark apparatus for 4 hours after which time an extra amount of sodium hydride (10 mg) was added and the reaction was refluxed for a further for 4 hours. After allowing to cool to room temperature, the toluene was washed with water, dried and evaporated. The residue was purified by column chromatography eluting with ethyl acetate / z^'søhexane / triethylamine (50 / 50 / 1) then ethyl acetate / triethylamine (99 / 1) to afford the sub-titled compound (0.200 g).

m/e 346 [M+H]⁺

¹H NMR (400 MHz, CDCl₃) δ 7.26 (td, IH), 7.10 - 7.07 (m, IH), 7.04 (dd, IH), 6.90 (ddd, IH), 4.78 - 4.73 (m, IH), 3.14 (ddd, IH), 2.79 - 2.66 (m, 3H), 2.66 - 2.56 (m, IH), 2.53 - 2.46 (m, IH), 2.46 - 2.36 (m, 2H), 2.13 - 1.99 (m, 2H), 1.90 - 1.85 (m, IH), 1.73 - 1.40 (m, HH), 1.29 - 1.18 (m, IH).

Example 8: (if)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(pyrazin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide

H

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

m/e 481 [M⁺] ¹H NMR (400 MHz, DMSO-D₆) δ 11.42 (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, 2H), 4.16 - 4.07 (m, IH), 3.71 - 3.57 (m, 4H), 3.52 - 3.41 (m, IH), 2.43 - 2.27 (m, 2H), 2.26 - 2.19 (m, IH), 2.19 - 2.09 (m, IH), 2.05 - 1.87 (m, 3H), 1.86 - 1.76 (m, IH), 1.71 - 1.46 (m, 9H).

Example 9: (if)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(isoxazol-3- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2] octane bromide

a) 2-Bromo-7V-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 with stirring and the suspension was stirred overnight. The reaction was washed with water (2 x 50 mL), dried and evaporated. The product was recrystallised from dichloromethane / z^'søhexane to afford the sub-titled compound (2.3 g).

¹H NMR POO MHZ₅ CDCI₃) O S^ (S, IH), 8.34 (S, IH), 7.06 (S, IH), 4.03 (S, 2H).

H l-(3-Fluoro-phenyl)-cycloheptanecarboxylic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 8d) (50 mg) and 2-bromo-7V-isoxazol-3-yl-acetamide (Example 9a) (30 mg) were dissolved in acetonitrile (4 mL) and stirred overnight. The solution was diluted with 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 mg).

m/e 470 [M⁺]

¹H NMR (400 MHz, DMSO-D₆) δ 11.69 (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, IH), 3.68 - 3.53 (m, 4H), 3.43 (dd, IH), 2.42 - 2.27 (m, 2H), 2.25 - 2.19 (m, IH), 2.18 - 2.09 (m, IH), 2.04 - 1.88 (m, 3H), 1.85 - 1.75 (m, IH), 1.69 - 1.51 (m, 9H).

Example 10: (if)-l-[(5-Fluoro-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo [2.2.2] octane chloride

a) 2-Chloro-7V-(5-fluoro-pyridin-2-yl)-acetamide

The title compound (0.99 g, 73%, white solid) was prepared according to the method used in Example 3f using 2-amino-5-fluoro-pyridine.

¹H NMR (400 MHz, DMSO-D₆) δ 10.91 (s, IH), 8.35 (d, IH), 8.10 (dd, IH), 7.80-7.74 (m, IH), 4.34 (s, 2H).

H

2-Chloro-7V-(5-fluoro-pyridin-2-yl)-acetamide (Example 10a) (31 mg) was added to a solution of 1-phenyl-cycloheptanecarboxylic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example 3e) (49 mg) in acetonitrile (1 mL). The reaction mixture was stirred at room temperature overnight. Diethyl ether (2 mL) was added to the reaction mixture and the white solid was filtered off, washed several times with diethyl ether and dried under vacuum at 40⁰C to give the title compound (49 mg).

m/e 480 [M]⁺ 1H NMR (400 MHz, DMSO-D₆) δ 11.19 (s, IH), 8.36 (d, IH), 8.02 (m, IH), 7.81 (ddd, 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 (m, 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), 1.99- 1.82 (m, 3H), 1.78-1.65 (m, IH), 1.70-1.41 (m, 9H).

Example 11: (if)-l-[(2-Methyl-pyridin-4-ylcarbamoyl)-methyl]-3-(l-phenyl- cycloheptanecarbonyloxy)-l-azonia-bicyclo [2.2.2] octane chloride

a) 2-Chloro-7V-(2-methyl-pyridin-4-yl)-acetamide

The title compound (1.0 g) was prepared according to the method used in Example 3f using 4-amino-2-methylpyridine.

¹H NMR (400 MHz, DMSO-D₆) δ 10.64 (s, IH), 8.32 (d, IH), 7.44 (d, IH), 7.38-7.35 (m, IH), 4.30 (s, 2H), 2.42 (s, 3H). Example 11: (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 prepare Example 10. Further purification was achieved by silica gel chromatography eluting with 0-20% MeOH / dichloromethane to give the title compound as a white solid (57 mg).

m/e 476 [M]⁺ 1H NMR (400 MHz, DMSO-D₆) δ 11.32 (s, IH), 8.31 (d, IH), 7.43 (d, IH), 7.35-7.26 (m, 5H), 7.22-7.16 (m, IH), 5.09-5.04 (m, IH), 4.30 (dd, 2H), 4.09-4.01 (m, IH), 3.64- 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).

Example 12: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-3- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2] octane chloride

a) 2-Chloro-7V-pyridin-3-yl-acetamide

A mixture of 3-aminopyridine (350 mg) and sodium hydroxide (0.6 g) were dissolved in water (8 mL) and the reaction mixture was cooled in an ice bath. Chloroacetyl chloride (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 the organic layer was concentrated and purified by column chromatography, eluting with 0- 60% ethyl acetate / cyclohexane to give the title compound (0.1 Og) as a white solid.

¹H NMR (400 MHz, DMSO-D₆) δ 10.51 (s, IH), 8.73 (d, IH), 8.30 (dd, IH), 8.03 (ddd, IH), 7.40-7.35 (m, IH), 4.30 (s, 2H).

H The title compound (78 mg) was prepared by an analogous method to that used in Example 3 using 2-chloro-7V-pyridin-3-yl-acetamide in place of 2-bromo-7V-pyridin-2-yl-acetamide.

m/e 462 [M]⁺

¹H NMR (400 MHz, DMSO-D₆) δ 11.27 (s, IH), 8.76 (d, IH), 8.30 (dd, IH), 7.98 (ddd, IH), 7.37 (ddd, IH), 7.33-7.25 (m, 4H), 7.22-7.15 (m, IH), 5.07 (d, IH), 4.28 (dd, 2H), 4.11-4.03 (m, IH), 3.65-3.50 (m, 4H), 3.41-3.29 (m, IH), 2.37-2.21 (m, 2H), 2.19-2.05 (m, 2H), 1.97-1.83 (m, 3H), 1.78-1.66 (m, IH), 1.71-1.27 (m, 9H).

Example 13: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridazin-3- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2]octane bromide

a) 2-Bromo-7V-pyridazin-3 -yl-acetamide

To a suspension of pyridazin-3-ylamine (2.7 g) and dizsøpropylethylamine (6.3 mL) in dichloromethane (100 mL) at 0⁰C was added bromoacetic anhydride (9.0 g) in dichloromethane (10 mL) by dropwise addition. The mixture was stirred at 0°C for 0.5 hours and then allowed to warm to rt. The resulting suspension was filtered, washed with dichloromethane and dried to afford the sub-titled compound as a solid (2.0 g).

¹H NMR (400 MHz, DMSO-D₆) δ 11.51 (s, IH), 9.00 (dd, IH), 8.28 (dd, IH), 7.74 - 7.68 (m, IH), 4.15 (s, 2H).

Example 13: (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridazin-3- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2] octane bromide

H

1 -Phenyl-cycloheptanecarboxylic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Example Ie) (0.160 g) and 2-bromo-7V-pyridazin-3-yl-acetamide (Example 13a) (0.106 g) were dissolved in acetonitrile (1 mL) and left to stand overnight. The solvents were removed under reduced pressure and the residue purified by chromatography on silica eluting with methanol / dichloromethane (1 : 9) to afford the titled compound as a solid (180 mg).

m/e 463 [M]⁺

¹H NMR (400 MHz, DMSO-D₆) δ 11.68 (s, IH), 9.06 (dd, IH), 8.25 (d, IH), 7.79 (dd, IH), 7.39 - 7.30 (m, 4H), 7.27 - 7.21 (m, IH), 5.15 - 5.10 (m, IH), 4.34 (s, 2H), 4.16 - 4.06 (m, 2H), 3.69 - 3.56 (m, 4H), 3.46 - 3.36 (m, IH), 2.43 - 2.27 (m, 2H), 2.24 - 2.10 (m, 2H), 2.04 - 1.89 (m, 3H), 1.84 - 1.71 (m, IH), 1.68 - 1.45 (m, 8H). Example 14: (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-2- ylcarbamoylmethyO-l-azonia-bicycloβ.l.l] octane hemi-naphthalene-l,5-disulfonate Crystalline Form A

(i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane chloride (Example 3) (100 mg) in dichloromethane (4 mL) was shaken with an aqueous solution of naphthalene- 1,5 -disulfonic acid di sodium salt (73 mg in 2 mL Of H₂O). The organic phase was collected and the aqueous layer extracted with dichloromethane (4 mL). The combined organic layers were passed through a phase separation cartridge and the resulting solution evaporated to give a colourless oil. The residue was triturated with diethyl ether and the resulting solid collected by filtration, washed with diethyl ether and dried at 50⁰C under vacuum. The solid was dissolved in hot acetonitrile (1 mL) and then evaporated to give a foam, which was then dissolved in acetone (2 mL). The mixture was left to stand for 48 h during which crystallistion occurred. The resulting crystals were collected by filtration, washed with ice-cooled acetone and then dried at 50⁰C under vacuum to afford the titled compound as a white solid (67 mg).

m/e 462 [M]⁺ 1H NMR (400 MHz, DMSO-D₆): δ 11.06 (s, IH), 8.85-8.88 (d, IH), 8.40-8.36 (d, IH), 7.98-8.06 (d, IH), 7.91-7.94 (dd, IH), 7.90-7.85 (dd, IH), 7.42-7.36 (dd, lH),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, 2H), 1.99-1.81 (m, 3H), 1.78- 1.66 (m, IH), 1.77-1.19 (m, 9H). Analysis of Example 14: (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-2- ylcarbamoylmethyl)-l-azonia-bicyclo[2.2.2] octane hemi-naphthalene-l,5-disulfonate 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 or CubiX system) and DSC.

The melting temperature of Example 14 hemi-naphthalene-l,5-disulfonate Form A as determined by DSC was found to be 198°C (onset) (±2°C). GVS determination gave a 1% weight increase (%w/w) at 80% RH (±0.3%).

An XRPD spectrum of Example 14 hemi-naphthalene-l,5-disulfonate Form A is presented in Figure 8.

Alternative preparation of (if)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridine-2- ylcarbamoylmethyl) -1-azoniabicyclo [2.2.2] octane bromide (Example 4)

General Conditions: Unless otherwise stated all reactions were carried out under an inert atmosphere (nitrogen); reagents and solvents were obtained commercially and used as received; reagent grade solvents were used

(a) Cycloheptanecarboxylic Acid Methyl Ester

Cycloheptanecarboxylic acid (3.75kg) and methanol (37.50 L) were charged to a reaction vessel and the resultant mixture stirred. Sulfuric Acid (100%, 51.73 g) was charged, the temperature raised to 60⁰C and stirring continued for 18 hours. Methanol was removed by distillation under reduced pressure to leave a total volume of 11.25L. Toluene (37.50 L) was charged and a further 15L of solvent removed by distillation under reduced pressure. Analysis by ¹H NMR spectroscopy was carried out to confirm that methanol was no longer present in the solution. The mixture was allowed to cool to ambient temperature and diluted with toluene (7.50 L). Saturated aqueous sodium bicarbonate (18.75L) was charged. The reaction mixture was stirred for 15 min, then stirring stopped and the layers allowed to separate. The lower aqueous layer was removed to waste. Saturated aqueous sodium chloride (18.75 L) was charged. The reaction mixture was stirred for 15 min, then stirring stopped and the layers allowed to separate. The lower aqueous layer was removed to waste. The crude product solution was dried by azeotropic distillation under reduced pressure to remove 7.5L of toluene, giving 28.3kg of a 14.08% w/w toluene solution of cycloheptanecarboxylic acid methyl ester.

(b) 1-Phenyl-cycloheptanecarboxylic Acid Methyl Ester

Diisopropylamine (3.44 kg) and toluene (16.52 kg) were charged to a first reaction vessel and cooled to 0⁰C with stirring. N-Hexyllithium (8.81 kg, 33%w/w) was added, maintaining a temperature of 5 ⁰C ± 5°C. The mixture was stirred for 20min at this temperature. Cycloheptanecarboxylic acid methyl ester (14.08% w/w in toluene; 26.93 kg) was first concentrated by removal of 11.37L of toluene by distillation under reduced pressure, then charged to the first reaction vessel, maintaining a temperature of 5 ⁰C ± 5°C. The contents were allowed to warm to 20⁰C, stirred for 20min at this temperature, then cooled back to 0⁰C. To a second reaction vessel was charged dibromo bis(tri-tert- butylphosphine) dipalladium (I) (Johnson Matthey Pd-113; 189.15 g); bromobenzene (3.06 L) and toluene (7.58 L) under an inert atmosphere at ambient temperature. The contents of the second vessel were charged to first vessel at a rate such that a temperature of 5°C ±

5°C, was maintained, followed by line wash of toluene (3.79 L). The mixture was stirred at 0⁰C for 1 hour, then allowed to warm to 20⁰C and stirred at this temperature overnight. 2M hydrochloric acid (18.96 L) was added, whilst maintaining the temperature below 30 ⁰C, then the mixture was stirred at 20 ⁰C for 15 min, stirring stopped and the layers allowed to separate. The lower aqueous layer was removed to waste. A second charge of 2M hydrochloric acid (18.96 L) was added, then the mixture stirred at 20 ⁰C for 15 min, stirring stopped and the layers allowed to separate. The lower aqueous layer was removed to waste. Water (18.96 L) was charged, the mixture stirred at 20 ⁰C for 15 min, then stirring stopped and the layers allowed to separate. The lower aqueous layer was removed to waste. The crude product solution was passed through cartridges containing

Phosphonics SPM32 scavenger, then evaporated to dryness under reduced pressure on a rotating film evaporator to give 1-Phenyl-cycloheptanecarboxylic Acid methyl ester as a mobile brown oil (3.12 kg)

(c) 1-Phenyl-cycloheptanecarboxylic Acid Sodium Hydroxide (12.64 kg) was dissolved in water (31.60 L) and cooled to 20⁰C. 1- Phenyl-cycloheptanecarboxylic acid methyl ester (6.32 kg) in methanol (31.60 L) was added followed by a line rinse of methanol (5 L). The mixture was stirred at 60⁰C for 18 hours, then cooled to 20⁰C. Concentrated hydrochloric acid (29.43 L) was added to precipitate the product, maintaining the temperature at below 50 ⁰C, then the mixture cooled to 20⁰C and stirred for 18 hours. The crude product was collected by filtration and washed with water (31.60 L), then dispersed in methanol (37.41 L) and water (9.35 L). The mixture was heated with stirring to 62 ⁰C at a rate of 1 °C/min then cooled to 5 ⁰C at a rate of 0.3 °C/min and held at 5 ⁰C overnight. The product was collected by filtration, washed with water (2 x 12.64 L) and dried in a vacuum oven at 40⁰C for 72 hours to give 1- Phenyl-cycloheptanecarboxylic acid (5.60 kg).

(d) 1-Phenyl-cycloheptanecarboxylic Acid (R)-(l-aza-bicyclo [2.2.2] oct-3-yl) ester

1 -Phenyl-cycloheptanecarboxylic acid (2.70 kg) and butanenitrile (21.60 L). were charged to a first reaction vessel. The contents were heated to 70±5°C with stirring to give a homogeneous solution. To a second reaction vessel was charged l,l'-carbonyldiimidazole (1.1 equiv (molar); 2.16 kg) and butanenitrile (10.80 L). The contents were heated to 50±5°C with stirring. The contents of the first vessel were transferred to the second vessel, the temperature raised to 70±5°C and stirring continued for 30±15min. (i?)-(-)-3- quinuclidinol (1.67 kg) and butanenitrile (8.10 L), were charged to the first reaction vesel followed by potassium-t-amylate (7.32 L). This mixture was stirred for 15 min, then added to the second vessel, followed by a line rinse of butanenitrile (1.35 L). The mixture was stirred at 70⁰C for 18 hours then cooled to 20⁰C. 1 M Hydrochloric acid (29.70 L) was charged, followed by sufficient concentrated hydrochloric acid to reduce the pH to below 7 (2.276kg added). The mixture was stirred for 15min, stirring stopped and the layers allowed to separate. The lower layer was removed to waste. Saturated aqueous sodium bicarbonate solution (27.00 L) was charged. The mixture was stirred for 15min, stirring stopped and the layers allowed to separate. The lower layer was removed to waste. Solvent was removed by distillation under reduced pressure to give a 31.9% w/w solution of the sub-title product (10.73kg of solution and 3.42kg of product).

(e) (R)-3-(l-Phenyl-cycloheptanecarbonyloxy)-l-(pyridin-2-ylcarbamoylmethyl)- 1-azonia-bicyclo [2.2.2] octane bromide

To bromoacetic acid (3.00 kg) in methyl acetate (24.18 L) in a first reaction vessel was charged 1-propanephosphonic acid cyclic anhydride (T3P) (19.51 L) in methyl acetate (50% w/w solution). The contents were cooled to 5°C with stirring, then a solution of 2- pyridinamine (8.13 kg) in methyl acetate (24.29 L) precooled to 10⁰C, was charged, keeping the temperature of the vessel contents below 5°C. The mixture was stirred for lhr, then stiring stopped and the layers allowed to separate. The lower layer was separated to waste. The remaining solution containing 2-bromo-7V-pyridin-2-yl-acetamide (42.04kg at 3.90% w/w) was transferred to a second reaction vessel and cooled to 0⁰C.

1-Phenyl-cycloheptanecarboxylic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (31.9% w/w solution in butanenitrile; 7.88 kg) was charged to the second reaction vessel and the mixture stirred at 0⁰C for 18 hours. The solid product was collected by filtration, washed with methyl acetate (6.22 L) and dried in a vacuum oven at 50⁰C to give crude product (3.51 kg at 90.5% w/w 3.17 Kg overall).

Crude Product (3.48 kg) in ethanol (69.62 L) was heated to 75°C until fully dissolved. The solution was filtered through a 1.2 micron filter, then cooled at a rate of 0.3⁰C / min to 0⁰C and stirred at this temperature for 18 hours. The solid product was collected by filtration, washed with ethanol (7.47 L) and dried in a vacuum oven at 50⁰C for 48 hours to give purified product (2.82 kg).

Biological Activity of Muscarinic Antagonists

The inhibitory effects of compounds of the muscarinic antagonists were determined by a Muscarinic Receptor Radioligand Binding Assay.

Radioligand binding studies utilising [³H]-N-methyl scopolamine ([³H]-NMS) and commercially available cell membranes expressing the human muscarinic receptors (M2 or M3) were used to assess the affinity of muscarinic antagonists for M2 and M3 receptors. Membranes in TRIS buffer were incubated in 96-well plates with [³H]-NMS and M3 antagonist at various concentrations for 3 hours. Membranes and bound radioligand were then harvested by filtration and allowed to dry overnight. Scintillation fluid was then added and the bound radioligand counted using a Canberra Packard Topcount scintillation counter

The half-life of antagonists at each muscarinic receptor was measured using the alternative radioligand [³H]-QNB and an adaptation of the above affinity assay. Antagonists were incubated for 3 hours at a concentration 10-fold higher than their Ki, as determined with the [³H]-QNB ligand, with membranes expressing the human muscarinic receptors. At the end of this time, [³H]-QNB was added to a concentration 25 -fold higher than its Kd for the receptor being studied and the incubation continued for various time periods from 15 minutes up to 180 minutes. Membranes and bound radioligand were then harvested by filtration and allowed to dry overnight. Scintillation fluid was then added and the bound radioligand counted using a Canberra Packard Topcount scintillation counter. The rate at which [³H]-QNB is detected binding to the muscarinic receptors is related to the rate at which the antagonist dissociates from the receptor, i.e. to the half life of the antagonists on the receptors. Table 1 shows the pICso figures for Example 1.

Table 1

Table 2 gives IC50 strengths for the compounds of the examples. Table 2

M3 Binding IC₅₀ <2nM "+++"; IC₅₀2-1OnM "++"; IC₅₀ > 1OnM "+"; NT - Not Tested.

Protocols for Combination Experiments 1. Evaluation of compound activity on isolated tracheal rings from guinea-pig preconstricted with methacholine.

The following protocol may be used to evaluate the effects of muscarinic M3 receptor antagonists according to the present invention in combination with budesonide.

Male albino Dunkin Hartley guinea-pigs (300-350 g) are killed by cervical dislocation and the trachea excised. Adherent connective tissue is removed and the trachea cut into ring segments (2-3 mm wide). These are suspended in 10ml organ baths containing a modified Krebs solution composition (mM): NaCl 117.56, KCI 5.36, NaH₂PO₄ 1.15, MgSO₄ 1.18, glucose 11.10, NaHCO₃ 25.00 and CaCI₂ 2.55. This is maintained at 37°C and continually gassed with 5% CO₂ in O₂, Indomethacin (2.8 μM), corticosterone (10 μM), ascorbate (1 mM), CGP20712A (1 μM) and phentolamine (3 μM) are added to the Krebs solution: indomethacin to prevent development of smooth muscle tone due to the synthesis of cyclooxygenase products, corticosterone to inhibit the uptake 2 process, ascorbate to prevent catecholamine oxidation and CGP20712A and phentolamine to avoid any complicating effects of βl- and α-adrenoceptor activation respectively. The tracheal rings are suspended between two stainless steel hooks, one attached to an isometric force transducer and the other to a stationary support in the organ bath. Changes in isometric force are recorded.

Acetyl-β-methylcholine chloride (Methacholine), Indomethacin, Corticosterone-21 -acetate, Phentolamine hydrochloride, Ascorbic acid, and CGP20712A methanesulphate may be obtained from the Sigma chemical company. Indomethacin may be dissolved in 10% w/v Na₂CO₃, corticosterone 21 -acetate in ethanol and other compounds in DMSO. Muscarinic antagonists and Budesonide may be diluted in Krebs prior to adding to tissues and the level of DMSO in the bath < 0.1 %.

At the beginning of each experiment a force of 1.0 g.wt. is applied to the tissues and this is reinstated over a 30min equilibration period until it remained steady. Tissues are then exposed to lμM of the muscarinic agonist, methacholine, to assess tissue viability. Tissues are washed by exchanging the bathing Krebs solution three times. After 30 minutes the tissues are precontracted with lμM methacholine. When the contraction reaches a plateau, 10OnM Budesonide, 1OnM Muscarinic antagonist or a combination of the two is added to the bathing media and left for 60 minutes.

Data may be collected using the AD Instruments Chart5 for windows software, the tension generated may be measured before addition of methacholine and after its response reaches a plateau. The response to the muscarinic antagonist and/or Budesonide may be measured at 10 minute intervals following their addition. All responses may be expressed as percentage inhibition of the methacholine-induced contraction.

2. Inflammatory Cell influx experiment in LPS-Challenged Rats

The following protocol may be used to evaluate the effects of muscarinic M3 receptor antagonists according to the present invention, in combination with CCRl anatgonists.

The effect of a CCRl receptor antagonist and a muscarinic antagonist according to the invention, and their combination, on inflammatory cell influx can be assayed by monitoring the effect on total cell number in broncholalveolar lavage (BAL) fluid of rats challenged intra-tracheally (i.t.) with Lipopolyaccharide (LPS) [N = 10 rats per treatment group].

Methodology LPS instillation: Rats are anaesthetized with Efrane and put in a supine position, head up, on a board tilted at 30°. LPS (Lipopolysaccharide B.E.coli 026:B6) (2.5 μg/ml) is dissolved in saline (0.9% NaCl), or saline alone (negative control) in a volume of 200 μl and administered i.t. using a modified metal cannula. Rats remain in this position until regaining consciousness.

Preparation of solutions: CCRl anatgonists are dissolved in 0.9% NaCl solution to a final concentration of 0.001 to 0.100 mg. Muscarinic antagonists are dissolved in 0.9% NaCl solution to an appropriate final concentration of 0.001 to 1.0 mg/ml. CCRl antagonist, Muscarinic antagonist or mixed s are made by dissolving CCRl antagonist in Muscarinic antagonist suspensions, giving a final concentration of 001 to 0.100 CCRl antagonist /ml and 001 to 1.0 mg Muscarinic antagonist /ml.

Treatments: Animals were intratracheally instilled with solutions (1 ml/kg) of Muscarinic antagonist / CCRl antagonist (0.002/ 001 to 0.100 mg/kg), or of Muscarinic antagonist (001 to 1.0 mg/kg) alone, or CCRl antagonist (001 to 0.100 mg/kg) alone, or with saline (negative and positive control animals). The treatments were carried out under light anaesthesia (Efrane) to secure that the solution reached the lungs. The drugs were administrated 30 min before the LPS instillation.

Termination: 4 hours after the LPS challenge, rats are intraperitoneally injected with the mixture (0.3 ml) of pentobarbital (60 mg/ml, Apoteksbolaget, Sweden) and PBS (1:1) for 1 - 2 min.

Bronchoalveolar lavage: After termination, BAL is performed twice with PBS. The BAL fluid is centrifuged and the cell pellet was resuspended in PBS. The total numbers of BAL cells is counted in a SYSMEX cell counter.

Claims

C L A I M S

1. A pharmaceutical product comprising, in combination, a first active ingredient which is a muscarinic antagonist selected from: (i?)-3-(l -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyrazin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane X

(i?)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(pyrazin-2-ylcarbamoylmethyl)- l-azonia-bicyclo[2.2.2]octane X;

(i?)-3-[l-(3-Fluoro-phenyl)-cycloheptanecarbonyloxy]-l-(isoxazol-3-ylcarbamoylmethyl)- l-azonia-bicyclo[2.2.2]octane X;

(i?)-3 -(I -Phenyl-cycloheptanecarbonyloxy)- 1 -(pyridin-2-ylcarbamoylmethyl)-l -azonia- bicyclo[2.2.2]octane X; (i?)-l-[(5-Fluoro-pyridin-2-ylcarbamoyl)-methyl]-3-(l-phenyl-cycloheptanecarbonyloxy)- l-azonia-bicyclo[2.2.2]octane X;

(R)- 1 -[(2-Methyl-pyridin-4-ylcarbamoyl)-methyl]-3-(l -phenyl-cycloheptanecarbonyloxy)- l-azonia-bicyclo[2.2.2]octane X; wherein X represents a pharmaceutically acceptable anion of a mono or polyvalent acid, and a second active ingredient which selected from i) a phosphodiesterase inhibitor, ii) a modulator of chemokine receptor function iii) an inhibitor of kinase function, iv) a protease inhibitor, v) a steroidal glucocorticoid receptor agonist, vi) a non-steroidal glucocorticoid receptor agonist, and vii) a purinoceptor antagonist.

2. A product according to claim 1 wherein the first active ingredient is a muscarinic antagonist, which is a 2,5-dichlorobenzene sulphonate or l-hydroxynaphthalene-2- sulphonate salt.

3. A product according to claim 1 wherein the first active ingredient is a muscarinic antagonist which is a l-hydroxynaphthalene-2-sulphonate salt.

4. A product according to any one of claims 1 to 3, wherein the second active ingredient is a CCRl antagonist.

5. A product according to claim 4 wherein the second active ingredient is a CCRl antagonist, selected from:

N- {2-[((25)-3- {[ 1 -(4-chlorobenzyl)piperidin-4-yl] amino} -2-hydroxy-2-methylpropyl)oxy]- 4-hydroxyphenyl} acetamide; N- {5-chloro-2-[((25)-3- {[ 1 -(4-chlorobenzyl)piperidin-4-yl] amino } -2-hydroxy-2- methylpropyl)oxy]-4-hydroxyphenyl}acetamide;

6. A product according to claim 5, wherein the second active ingredient is N-{2-[((2S)-3- {[l-(4-chlorobenzyl)piperidin-4-yl]amino}-2-hydroxy-2-methylpropyl)oxy]-4- hydroxyphenyl} acetamide or a pharmaceutically acceptable salt thereof.

7. A product according to claim 5, wherein the second active ingredient is 7V-{5-chloro-2- [((25)-3- {[ 1 -(4-chlorobenzyl)piperidin-4-yl] amino} -2-hydroxy-2-methylpropyl)oxy]-4- hydroxyphenyl} acetamide or a pharmaceutically acceptable salt thereof.

8. A product according to claim 5, wherein the second active ingredient is 2-{2-chloro-5- {[(2S)-3-(5-chloro-lΗ,3H-spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2- hydroxypropyl]oxy}-4-[(methylamino)carbonyl]phenoxy} -2-methylpropanoic acid or a pharmaceutically acceptable salt thereof.

9. A product according to any one of claims 1 to 3, wherein the second active ingredient is a steroidal glucocorticoid receptor agonist.

10. Use of a product according to any one of claims 1 to 9 in the manufacture of a medicament for the treatment of a respiratory disease.

11. Use according to claim 10, wherein the respiratory disease is chronic obstructive pulmonary disease.

12. A method of treating a respiratory disease, which method comprises simultaneously, sequentially or separately administering:

(a) a (therapeutically effective) dose of a first active ingredient which is a muscarinic receptor antagonist as defined in any one of claims 1 to 3; and (b) a (therapeutically effective) dose of a second active ingredient as defined in claim 1; to a patient in need thereof.

13. A kit comprising a preparation of a first active ingredient which is a muscarinic receptor antagonist as defined in any one of claims 1 to 3, and a preparation of a second active ingredient as defined in claim 1 and optionally instructions for the simultaneous, sequential or separate administration of the preparations to a patient in need thereof.

14. A pharmaceutical composition comprising, in admixture, a first active ingredient which is a muscarinic receptor antagonist as defined in any one of claims 1 to 3 and a second active ingredient as defined in claim 1.