WO2010004319A1

WO2010004319A1 - Combination comprising 6-flu0r0-n- ((1s, 4s) - 4- (6-fluoro-2, 4-di0x0-1- (4'- (piperazin-1- ylmethyl) biphenyl- 3-yl) -1, 2-dihydropyrido [2, 3-d] pyrimidin-3 (4h) - yl) cyclohexyl) imidazo [1,2-a] pyridine -2- carboxamide or a salt

Info

Publication number: WO2010004319A1
Application number: PCT/GB2009/050790
Authority: WO
Inventors: Paul Hartopp; Timothy Johnson; Hitesh Jayantilal Sanganee
Original assignee: Astrazeneca Ab; Astrazeneca Uk Limited
Priority date: 2008-07-07
Filing date: 2009-07-06
Publication date: 2010-01-14

Abstract

The invention provides a pharmaceutical product, kit or composition comprising a first active ingredient which is 6-fluoro-N-((1s,4s)-4-(6-fluoro-2,4-dioxo-1-(4'-(piperazin-1- ylmethy1)-biphenyl-3-yl)-l,2-dihydropyrido[2,3-d]pyrimidin-3(4H)- yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide or a salt thereof, and a second active ingredient selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; an antioxidant; a β₂ adrenoceptor agonist; a CCR1 antagonist; a chemokine antagonist (not CCR1); a corticosteroid; a CRTh2 antagonist; a DP1 antagonist; an Histone Deacetylase Inducer; an IKK2 inhibitor; a COX inhibitor; a lipoxygenase inhibitor; a leukotriene receptor antagonist; a MABA compound; an MPO inhibitor; a muscarinic antagonist; a p38 inhibitor; a PPARγ agonist; a protease inhibitor; a Statin; a thromboxane antagonist; a vasodilator; or, an ENAC blocker (Epithelial Sodium-channel blocker); and its use in the treatment of respiratory disease.

Description

COMBINATION COMPRISING 6-FLU0R0-N- ((1S, 4S) - 4- (6-FLUORO-2 , 4-DI0X0-1-

(4'-(PIPERAZIN-1- YLMETHYL) BIPHENYL- 3-YL) -1, 2-DIHYDROPYRIDO [2, 3-D]

PYRIMIDIN-3(4H) - YL) CYCLOHEXYL) IMIDAZO [1,2-A] PYRIDINE -2-

CARBOXAMIDE OR A SALT

The present invention relates to a combination of two or more pharmaceutically active substances for use in the treatment of respiratory diseases (for example chronic obstructive pulmonary disease (COPD) or 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 corticosteroids. Corticosteroids (also known as glucocorticosteroids or glucocorticoids) are potent anti-inflammatory agents. Whilst their exact mechanism of action is not clear, the end result of corticosteroid treatment is a decrease in the number, activity and movement of inflammatory cells into the bronchial submucosa, leading to decreased airway responsiveness. Corticosteroids may also cause reduced shedding of bronchial epithelial lining, vascular permeability, and mucus secretion. Whilst corticosteroid treatment can yield important benefits, the efficacy of these agents is often far from satisfactory, particularly in COPD. Moreover, whilst the use of steroids may lead to therapeutic effects, it is desirable to be able to use steroids in low doses to minimise the occurrence and severity of undesirable side effects that may be associated with regular administration. Recent studies have also highlighted the problem of the acquisition of steroid resistance amongst patients suffering from respiratory diseases. For example, cigarette smokers with asthma have been found to be insensitive to short term inhaled corticosteroid therapy, but the disparity of the response between smokers and non-smokers appears to be reduced with high dose inhaled corticosteroid (Tomlinson et al., Thorax 2005;60:282-287).

A further class of therapeutic agent used in the treatment of respiratory diseases are bronchodilators. Bronchodilators may be used to alleviate symptoms of respiratory diseases by relaxing the bronchial smooth muscles, reducing airway obstruction, reducing lung hyperinflation and decreasing shortness of breath. Types of bronchodilators in clinical use include β₂ adrenoceptor agonists, muscarinic receptor antagonists and methylxanthines. Bronchodilators are prescribed mainly for symptomatic relief and they are not considered to alter the natural history of respiratory diseases. 6-Fluoro-N-(( 1 s,4s)-4-(6-fluoro-2,4-dioxo- 1 -(4'-(piperazin- 1 -ylmethyl)-biphenyl-3 - yl)-l,2-dihydropyrido[2,3-d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2- carboxamide and many of its salts (such as the (lS)-(+)-10-Camphorsulfonic acid salt) are inhibitors of PDE4 and are disclosed in PCT/GB2008/000061. Combination products comprising a β₂ adrenoceptor agonist and a corticosteroid are available. One such product is a combination of budesonide and formoterol fumarate

(marketed by AstraZeneca under the tradename Symbicort ®), which has proven to be effective in controlling asthma and COPD, and improving quality of life in many patients.

In view of the complexity of respiratory diseases such as asthma and COPD, it is unlikely that any one mediator can satisfactorily treat a respiratory disease alone.

Moreover, whilst combination treatments using a β₂ adrenoceptor agonist and a corticosteroid deliver significant patient benefits, there remains a medical need for new therapies against respiratory diseases such as asthma and COPD, in particular for therapies with disease modifying potential. Accordingly, the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 6-fluoro-N-((ls,4s)-4-(6-fluoro-2,4-dioxo-l-

(4'-(piperazin-l-ylmethyl)-biphenyl-3-yl)-l,2-dihydropyrido[2,3-d]pyrimidin-3(4H)- yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide or a salt thereof, and a second active ingredient selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; an antioxidant; a β₂ adrenoceptor agonist; a CCRl antagonist; a chemokine antagonist (not CCRl); a corticosteroid; a CRTh2 antagonist; a DPI antagonist; an Histone Deacetylase Inducer; an IKK2 inhibitor; a COX inhibitor; a lipoxygenase inhibitor; a leukotriene receptor antagonist; a MABA compound; an MPO inhibitor; a muscarinic antagonist; a p38 inhibitor; a PPARγ agonist; a protease inhibitor; a Statin; a thromboxane antagonist; a vasodilator; or, an ENAC blocker (Epithelial Sodium-channel blocker).

The first active ingredient, which is 6-fluoro-N-((ls,4s)-4-(6-fluoro-2,4-dioxo-l-(4'- (piperazin- 1 -ylmethyl)-biphenyl-3-yl)- 1 ,2-dihydropyrido[2,3-d]pyrimidin-3(4H)- yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide or a salt thereof, may be in the form of a solvate (such as a hydrate). A suitable salt of 6-fluoro-N-((l s,4s)-4-(6-fluoro-2,4-dioxo- 1 -(4'-(piperazin- 1 - ylmethyl)-biphenyl-3-yl)-l,2-dihydropyrido[2,3-d]pyrimidin-3(4H)- yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide is, for example, a hydrochloride (such as a trihydrochloride), hydrobromide (such as dihydrobromide), trifluoroacetate, sulphate, phosphate, acetate, fumarate, maleate, tartrate, lactate, citrate, pyruvate, succinate, oxalate, methanesulphonate, /?-toluenesulphonate, bisulphate, benzenesulphonate, ethanesulphonate, malonate, xinafoate, ascorbate, oleate, nicotinate, saccharinate, adipate, formate, glycolate, L-lactate, D-lactate, aspartate, malate, L-tartrate, D-tartrate, stearate, 2-furoate, 3-furoate, napadisylate (naphthalene- 1, 5 -disulfonate or naphthalene- 1 -(sulfonic acid)-5-sulfonate), edisylate (ethane- 1,2 -disulfonate or ethane- 1- (sulfonic acid)-2-sulfonate), isethionate (2-hydroxyethylsulfonate), 2- mesitylenesulphonate, 2-naphthalenesulphonate, 2,5-dichlorobenzenesulphonate, D- mandelate, L-mandelate, cinnamate, benzoate, adipate, esylate, malonate, mesitylate (2- mesitylenesulphonate), napsylate (2-naphthalenesulfonate), camsylate (camphor- 10- sulphonate, for example (lS)-(+)-10-Camphorsulfonic acid salt), formate, glutamate, glutarate, glycolate, hippurate (2-(benzoylamino)acetate), orotate, xylate (p-xylene-2- sulphonate), pamoic (2,2'-dihydroxy-l,r-dinaphthylmethane-3,3'-dicarboxylate), palmitate or furoate.

In one aspect the present invention provides a pharmaceutical product wherein the first active ingredient is 6-fluoro-N-((ls,4s)-4-(6-fluoro-2,4-dioxo-l-(4'-(piperazin-l-ylmethyl)- biphenyl-3-yl)- 1 ,2-dihydropyrido[2,3-d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[ 1 ,2- a]pyridine-2-carboxamide as a (lS)-(+)-10-Camphorsulfonic acid or trihydrochloride salt. The first and second active ingredients can be administered simultaneously (either in a single pharmaceutical preparation {that is, the active ingredients are in admixture} or via separate preparations), or sequentially or separately via separate pharmaceutical preparations.

A non-steroidal glucocorticoid receptor (GR) agonist is, for example, a compound disclosed in WO 2006/046916.

An antioxidant is, for example, Allopurinol, Erdosteine, Mannitol, N-acetyl cysteine choline ester, N-acetyl cysteine ethyl ester, N-Acetylcysteine, N-Acetylcysteine amide or Niacin.

A β₂-adrenoceptor agonist is any compound or substance capable of stimulating the β₂ -receptors and acting as a bronchodilator. In the context of the present specification, unless otherwise stated, any reference to a β₂-adrenoceptor agonist includes an active salt, solvate or derivative that may be formed from said β₂-adrenoceptor agonist or any enantiomer or mixture thereof. Examples of possible salts or derivatives of β₂- adrenoceptor agonist are acid addition salts such as the salts of hydrochloric acid, hydrobromic acid, sulphuric acid, phosphoric acid, methanesulphonic acid, acetic acid, fumaric acid, succinic acid, lactic acid, citric acid, tartaric acid, l-hydroxy-2- naphthalenecarboxylic acid, maleic acid, and pharmaceutically acceptable esters (e.g. C₁- Ce alkyl esters). The β₂ -agonists may also be in the form of solvates, e.g. hydrates.

Examples of a β₂-adrenoceptor agonist that may be used in the pharmaceutical product according to this embodiment include metaproterenol, isoproterenol, isoprenaline, albuterol, salbutamol (e.g. as sulphate), formoterol (e.g. as fumarate), salmeterol (e.g. as xinafoate), terbutaline, orciprenaline, bitolterol (e.g. as mesylate), pirbuterol or indacaterol. The β₂-adrenoceptor agonist of this embodiment may be a long-acting β₂-agonist (i.e. a β₂- agonist with activity that persists for more than 24 hours), for example salmeterol (e.g. as xinafoate), formoterol (e.g. as fumarate), bambuterol (e.g. as hydrochloride), carmoterol (TA 2005, chemically identified as 2(1H)-Quinolone, 8-hydroxy-5-[l-hydroxy-2-[[2-(4- methoxy-phenyl)-l-methylethyl]-amino]ethyl]-monohydrochloride, [R-(R*, R*)] also identified by Chemical Abstract Service Registry Number 137888-11-0 and disclosed in U.S. Patent No 4,579,854), indacaterol (CAS no 312753-06-3; QAB-149), formanilide derivatives e.g. 3-(4-{[6-({(2R)-2-[3-(formylamino)-4-hydroxyphenyl]-2- hydroxyethyl}amino)hexyl]oxy}-butyl)-benzenesulfonamide as disclosed in WO

2002/76933, benzenesulfonamide derivatives e.g. 3-(4-{[6-({(2R)-2-hydroxy-2-[4- hydroxy-3-(hydroxy-methyl)phenyl]ethyl}amino)-hexyl]oxy}butyl)benzenesulfonamide as disclosed in WO 2002/88167, aryl aniline receptor agonists as disclosed in WO 2003/042164 and WO 2005/025555, indole derivatives as disclosed in WO 2004/032921, in US 2005/222144, compounds GSK 159797, GSK 159802, GSK 597901, GSK 642444 and GSK 678007.

In an embodiment of the present invention, the β₂-adrenoceptor agonist is 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 N-[2-hydroxy-5-[(lR)-l- hydroxy-2- [[( 1 R)-2-(4-methoxyphenyl)- 1 -methylethyl] amino] ethyljphenyl] -formamide, N-

[2-hydroxy-5-[(lS)-l-hydroxy-2-[[(lS)-2-(4-methoxyphenyl)-l- methylethyl]amino]ethyl]phenyl]-formamide and a mixture of such enantiomers, including a racemate.

In an embodiment of the invention, the β₂-adrenoceptor agonist is selected from: N-[2-(Diethylamino)ethyl]-N-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3-benzothiazol-7- yl)ethyl] amino } ethyl)-3 -[2-(I -naphthyl)ethoxy]propanamide;

N-[2-(Diethylamino)ethyl]-N-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3-benzothiazol-7- yl)ethyl] amino } ethyl)-3 - [2-(3 -chlorophenyl)ethoxy]propanamide;

7-[(\R)-2-( {2-[(3- {[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyl} amino)- 1 - hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3H)-one; and,

N-Cyclohexyl-N³-[2-(3-fluorophenyl)ethyl]-N-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3- benzothiazol-7-yl)ethyl] amino } ethyl)-β-alaninamide; or a pharmaceutically acceptable salt thereof. The names of the β2-adrenoceptor agonists of this embodiment are IUPAC names generated by the IUPAC NAME, ACD Labs Version 8 naming package.

In a further embodiment of the invention, the β₂-adrenoceptor agonist is selected from:

N-[2-(Diethylamino)ethyl]-N-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3-benzothiazol-7- yl)ethyl] amino } ethyl)-3 - [2-( 1 -naphthy l)ethoxy]propanamide dihydrobromide; N-[2-(Diethylamino)ethyl]-N-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3-benzothiazol-7- yl)ethyl] amino } ethyl)-3 - [2-(3 -chlorophenyl)ethoxy]propanamide dihydrobromide; 7-[(\R)-2-( {2-[(3- {[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyl} amino)- 1 - hydroxyethyl] -4 -hydroxy- 1 ,3-benzothiazol-2(3H)-one dihydrobromide; and, N-Cyclohexyl-Λ/³-[2-(3-fluorophenyl)ethyl]-N-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3- benzothiazol-7-yl)ethyl] amino } ethyl)-β-alaninamide di-D-mandelate salt.

A CCRl antagonist is, for example, a compound disclosed in WO2001/062728 or WO2001/098273, or a pharmaceutically acceptable salt thereof (such as a hydrochloride, trifluoroacetate, sulphate, (hemi)fumarate, benzoate, furoate or succinate salt); BX471 ((2R)- 1 - [ [2- [(aminocarbonyl)amino] -4-chlorophenoxy]acetyl]-4- [(4-fluorophenyl)methyl] - 2-methylpiperazine monohydrochloride) or CCX634.

Also, a CCRl antagonist is, for example, a compound disclosed in WO2001/062728 or WO2001/098273 [such as N-(2{(2S)-3[{(3R)-l-[(4- chlorophenyl)methyl]-3-pyrrolidinyl}amino]-2-hydroxypropoxy}-4- fiuorophenyl)acetamide, N-(2 {(2S)-3 [ ((3S)- 1 -[(4-chlorophenyl)methyl]-3- pyrrolidinyl} amino] -2-hydroxypropoxy} -4-fluorophenyl)acetamide, N-(2- {(2S)-3-[l - {(4- chlorobenzoyl)-4-piperidinyl} amino] -2-hydroxypropoxy} -4-hydroxyphenyl)acetamide, (2- {[(2S)-3-{[(2R,5S)-l-(4-chlorobenzyl)-2,5-dimethylpiperidin-4-yl]amino}-2-hydroxy-2- methylpropyl]oxy } -4-fluorophenyl)acetic acid, (2- { [(2S)-3 - { [(3 S ,4R)- 1 -(4-chlorobenzyl)- 3-methylpiperidin-4-yl]amino} -2-hydroxy-2-methylpropyl]oxy} -4-fluorophenyl)acetic acid, (2- { [(2S)-3- { [(3R,4R)- 1 -(4-chlorobenzyl)-3 -methylpiperidin-4-yl]amino} -2- hydroxy-2-methylpropyl]oxy}-4-fiuorophenyl)acetic acid, (2-{[(2S)-3-{[(2R,4S,5S)-l-(4- chlorobenzyl)-2,5-dimethylpiperidin-4-yl]amino}-2-hydroxy-2-methylpropyl]oxy}-4- fiuorophenyl)acetic acid, (2-{[(2S)-3-{[(2R,4R,5S)-l-(4-chlorobenzyl)-2,5- dimethylpiperidin-4-yl] amino} -2-hydroxy-2-methylpropyl]oxy} -4-fluorophenyl)acetic acid, (2-{[(2S)-3-{[(2S,4R,5R)-l-(4-chlorobenzyl)-2,5-dimethylpiperidin-4-yl]amino}-2- hydroxy-2-methylpropyl]oxy } -4-fluorophenyl)acetic acid, (2- { [(2S)-3- { [(2S,4S,5R)- 1 -(4- chlorobenzyl)-2,5-dimethylpiperidin-4-yl]amino}-2-hydroxy-2-methylpropyl]oxy}-4- fluorophenyl)acetic acid, Methyl (2-{[(25)-3-{[l-(4-chlorobenzyl)piperidin-4-yl]amino}-2- hydroxypropyl]oxy}-4-fluorophenyl)propanoate, N-[2-({2S}-3-[(l-[4-chlorobenzyl]-4- piperidinyl)amino]-2-hydroxypropoxy)-4-chlorophenyl acetamide, N-[2-({2S}-3-[(l-[4- chlorobenzyl]-4-piperidinyl)amino]-2-hydroxy-2-methylpropoxy)-4-hydroxyphenyl] acetamide, N-[2-({2S}-3-[(l-[4-chlorobenzyl]-4-piperidinyl)amino]-2-hydroxy-2- methylpropoxy)-4-fluorophenyl] acetamide, N-[5-chloro-[2-({2S}-3-[(l-[4-chlorobenzyl]- 4-piperidinyl)amino]-2-hydroxy-2-methylpropoxy)-4-hydroxyphenyl] acetamide, N-[5- chloro-[2-({2S}-3-[(l-[4-chlorobenzyl]-4-piperidinyl)amino]-2-hydroxy-2- methylpropoxy)-4-hydroxyphenyl] propaneamide, (2- { [(2S)-3- { [ 1 -(4- chlorobenzyl)piperidin-4-yl]amino}-2-hydroxy-2-methylpropyl]oxy}-4- fluorophenyl)methanesulfonic acid, N-5-chloro-(2- {(2S)-3-[ 1 - {(4-chlorobenzyl)-4- piperidinyl} amino]-2-hydroxypropoxy} -4-hydroxyphenyl)-N'-cyclopropyl-urea, N-(2- {(2S)-3-[ 1 - {(4-chlorobenzyl)-4-piperidinyl} amino] -2-hydroxypropoxy} -phenyl)-N' -ethyl- urea, (2S)-l-(2-ethylphenoxy)-3[(l-[4-chlorobenzyl]4-piperidinyl)amino]propan-2-ol, (2S)-l-[2-(-hydroxyethyl)phenoxy]-2-methyl-3[(l-[4-chlorobenzyl]-4- piperidinyl)amino]propan-2-ol, 2-({2S}-3-[(l-[4-chlorobenzyl]-4-piperidinyl)amino]-2- hydroxy-2-methylpropoxy)benzaldehyde, 2-({2S}-3-[(l-[4-chlorobenzyl]-4- piperidinyl)amino] -2-hydroxypropoxy)-N-cyclopropylbenzamide, Methyl 2-( {2S } -3 - [( 1 - [4-chlorobenzyl]-4-piperidinyl)amino]-2-hydroxypropoxy)-4-fluorobenzoate, N-(2-{[(2S)- 3-(5-chloro- 1 Η,3H-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy} -A- hydroxyphenyl)acetamide, N-(2-{[(2S)-3-(5-chloro-lΗ-spiro[l,3-benzodioxole-2,4'- piperidin]-r-yl)-2-hydroxypropyl]oxy}-4-hydroxyphenyl)acetamide, 2-{[(2S)-3-(5-chloro- 1 Η,3H-spiro[l -benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy} -4-hydroxy-N- methylbenzamide, 2- {[(2S)-3-(5-chloro-l'H,3H-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 ^?-yl)- 2-hydroxypropyl]oxy}-4-hydroxybenzoic acid, N-(2-{[(2S)-3-(5-chloro-l'H,3H-spiro[2- benzofuran-l,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-4-hydroxyphenyl)acetamide; 2-{[(2S)-3-(5-chloro-rH,3H-spiro[2-benzofuran-l,4'-piperidin]-r-yl)-2- hydroxypropyl]oxy}-4-hydroxy-N-methylbenzamide, N-(2-{[(2S)-3-(5-fluoro-l'H,3H- spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-4- hydroxyphenyl)acetamide, 2-{[(2S)-3-(5-fluoro-l'H,3H-spiro[l-benzofuran-2,4'- piperidin]-r-yl)-2-hydroxypropyl]oxy}-4-hydroxy-N-methylbenzamide, N-[2-({(2S)-3- [(2R)-5-chloro- 1 Η,3H-spiro[ 1 -benzofuran-2,3'-pyrrolidin]-r-yl]-2-hydroxypropyl}oxy)-4- hydroxyphenyljacetamide, N-(2-{[(2S)-3-(5-chloro-l'H,3H-spiro[l-benzofuran-2,4'- piperidin]-r-yl)-2-hydroxypropyl]oxy}-4-hydroxyphenyl)urea, 4-fluoro-2-{[(2S)-3-(5- fluoro-lΗ,3H-spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}benzoic acid, N-(2- { [(2S)-3-(5-chloro- 1 Η,3H-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-yl)-2- hydroxypropyl]oxy}-4-fluorophenyl)urea, N-(2-{[(2S)-2-amino-3-(5-fluoro-lΗ,3H- spiro[l -benzofuran-2,4'-piperidin]- 1 '-yl)propyl]oxy} -4-hydroxyphenyl)acetamide, 2-[(2S)- 3-(5-chlorospiro[benzofuran-2(3H),4'-piperidin]-r-yl)-2-hydroxypropoxy]-benzaldehyde, (αS)-5-chloro-α-[[2-(2-hydroxyethyl)phenoxy]methyl]-Spiro[benzofuran-2(3H),4'- piperidine]- 1 '-ethanol, (αS)-5-chloro-α-[[2-(hydroxymethyl)phenoxy]methyl]- Spiro[benzofuran-2(3H),4'-piperidine]- 1 '-ethanol, N-(2- {[(2S)-3-(5-chloro- 1 Η,3H-spiro[ 1 - benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy} -5-chloro-4- hydroxyphenyl)acetamide, 2-Chloro-5- {[(25)-3-(5-chloro- 1 '/f,3/f-spiro[ 1 -benzofuran-2,4'- piperidin]-r-yl)-2-hydroxypropyl]oxy}-(4-{acetylamino}phenoxy)acetic acid, 5-{[(2S)-3- (5-Chloro-r/f,3/f-spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-(4- {acetylamino}phenoxy)acetic acid, {2-Chloro-5- {[(25)-3-(5-chloro- 1 'H,3H-spiro[ 1 - benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy} -A- [(methylamino)carbonyl]phenoxy}acetic acid, 2-{2-Chloro-5-{[(25)-3-(5-chloro-l'H,3H- spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-4- [(methylamino)carbonyl]phenoxy}-2-methylpropanoic acid, (2-Chloro-5-{[(25)-3-(5- chloro- 1 '/f,3/f-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy } -A- { [(35)-3- hydroxypyrrolidin-l-yl]carbonyl}phenoxy)acetic acid, 5-Chloro-2-{[(25)-3-(5-chloro- r/f,3/f-spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-4-

(cyanomethoxy)benzoic acid, 2- {[(2S)-3-(5-chloro- 1 Η,3H-spiro[ 1 -benzofuran-2,4'- piperidin]-r-yl)-2-hydroxypropyl]oxy}-5-chloro-4-(2,2-difluoroethoxy)benzoic, 5-Chloro- 2- {[(25)-3-(5-chloro-r/f,3/f-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-yl)-2- hydroxypropyl]oxy}-4-(3,3,3-trifluoropropoxy)benzoic acid, N-(2-{3-[5-chloro-l'H,3H- spiro[l-benzofuran-2,4'-piperidin]-r-yl]propoxy}phenyl)acetamide, Methyl 3-(2-{[(2S)-3- (5-chloro- 1 'H3/f-spiro[ 1 -benzofuran-2,4 ' -piperidin]- 1 ' -yl)-2-hydroxypropyl]oxy } -4- fluorophenyl)propanoic acid, N-(2-{[(2S)-3-({spiro[indole-2-4'-piperidin]-3(lH)-one}-r- yl)-2-hydroxypropyl]oxy}-4-hydroxyphenyl)acetamide, or (2-{[(25)-3-(5-Chloro-r/f,3/f- spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-4- fluorophenyl)methanesulfonic acid, or a pharmaceutically acceptable salt thereof (for example as described above; (such as a hydrochloride, trifluoroacetate, sulphate, (hemi)fumarate, benzoate, furoate or succinate salt))]; BX471 ((2R)-l-[[2- [(aminocarbonyl)amino]-4-chlorophenoxy]acetyl]-4-[(4-fluorophenyl)methyl]-2- methylpiperazine monohydrochloride); or CCX634. Also, a CCRl antagonist is, for example, N-{2-[((2S)-3-{[\-(A- chlorobenzyl)piperidin-4-yl] amino } -2-hydroxy-2-methylpropyl)oxy] -A- hydroxyphenyl}acetamide (see WO 2003/051839), or, 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 (see PCT publication no. WO 2008/010765), or a pharmaceutically acceptable salt thereof (for example a hydrochloride, sulphate, (hemi)fumarate, benzoate, furoate or succinate salt).

A chemokine antagonist (other than a CCRl antagonist), for example, 656933 (N- (2-bromophenyl)-N'-(4-cyano- IH-1 ,2,3 -benzotriazol-7-yl)urea), 766994 (4-( {[({ [(2R)-4- (3,4-dichlorobenzyl)morpholin-2-yl]methyl}amino)carbonyl]-amino}methyl)benzamide), CCX-282, CCX-915, Cyanovirin N, E-921, INCB-003284, INCB-9471, Maraviroc, MLN- 3701, MLN-3897, T-487 (N-{l-[3-(4-ethoxyphenyl)-4-oxo-3,4-dihydropyrido[2,3- d]pyrimidin-2-yl] ethyl} -N-(pyridin-3 -ylmethyl)-2- [4-(trifluoromethoxy)phenyl] acetamide) or Vicriviroc. A corticosteroid is, for example, Alclometasone dipropionate, Amelometasone,

Beclomethasone dipropionate, Budesonide, Butixocort propionate, Ciclesonide, Clobetasol propionate, Desisobutyrylciclesonide, Etiprednol dicloacetate, Fluocinolone acetonide, Fluticasone Furoate, Fluticasone propionate, Loteprednol etabonate (topical) or Mometasone furoate. A CRTh2 antagonist is, for example, a compound from WO 2004/106302 or WO

2005/018529.

A DPI antagonist is, for example, L888839 or MK0525.

An histone deacetylase inducer is, for example, ADC4022, Aminophylline, a Methylxanthine or Theophylline. An IKK2 inhibitor is, for example, 2-{[2-(2-Methylamino-pyrimidin-4-yl)-lH- indole-5 -carbonyl] -amino } -3 -(phenyl-pyridin-2-yl-amino)-propionic acid.

A COX inhibitor is, for example, Celecoxib, Diclofenac sodium, Etodolac, Ibuprofen, Indomethacin, Meloxicam, Nimesulide, OC1768, OC2125, OC2184, OC499, OCD9101, Parecoxib sodium, Piceatannol, Piroxicam, Rofecoxib or Valdecoxib.

A lipoxygenase inhibitor is, for example, Ajulemic acid, Darbufelone, Darbufelone mesilate, Dexibuprofen lysine (monohydrate), Etalocib sodium, Licofelone, Linazolast, Lonapalene, Masoprocol, MN-OOl , Tepoxalin, UCB-35440, Veliflapon, ZD-2138, ZD- 4007 or Zileuton ((±)-l-(l-Benzo[b]thien-2-ylethyl)-l -hydroxyurea) A leukotriene receptor antagonist is, for example, Ablukast, Iralukast (CGP

45715A), Montelukast, Montelukast sodium, Ontazolast, Pranlukast, Pranlukast hydrate (mono Na salt), Verlukast (MK-679) or Zafirlukast.

A MABA compound is a compound having dual activity as both a muscarinic antagonist and as a β₂-adrenoceptor agonist, for example a MABA is a compound disclosed in: WO2004089892, WO2004106333, US20040167167, WO2005111004,

WO2005051946, WO2006023457, WO2006023460, US20060223858, US20060223859, WO2007107828, WO2008000483, US7317102 or WO2008041095. For example a MABA is: biphenyl-2-ylcarbamic acid l-[2-(4-{[(R)-2-(3-formylamino-4- hydroxyphenyl)-2-hydroxyethylam-2,5-dimethylphenylcarbamoyl)ethyl]piperidin-4-yl ester; or succinic acid salt and 1 ,2-ethanedisulfonic of biphenyl-2-ylcarbamic acid l-[2-(2- chloro-4-{[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-l,2-dihydroquinolin-5- yl)ethylmino]methyl} -5-methoxyphenyIcarbamoyl)ethyl]piperidin-4-yl ester; or naphthalene- 1, 5 -disulfonic acid salt of biphenyl-2-ylcarbamic acid l-(9-[(R)-2-hydroxy-2- (8-hydroxy-2-oxo-l ,2-dihydro-quinolin-5-yl)ethylamino]nonyl}piperidin-4-yl ester.

An MPO Inhibitor is, for example, a Hydroxamic acid derivative (N-(4-chloro-2- methyl-phenyl)-4-phenyl-4-[[(4-propan-2-ylphenyl)sulfonylamino]methyl]piperidine-l- carboxamide), Piceatannol or Resveratrol.

A muscarinic antagonist is, for example, Aclidinium bromide, Glycopyrrolate (such as R,R-, R,S-, S,R-, or S,S-glycopyrronium bromide), Oxitropium bromide, Pirenzepine, telenzepine, Tiotropium bromide, Darotropium ((1R, 3R, 5S)-3-(2-cyano-2,2- diphenylethyl)-8,8-dimethyl-8-azoniabicyclo[3,2,l]octane bromide), 3(R)-(2-hydroxy-2,2- dithien-2-ylacetoxy)-l-(3-phenoxypropyl)-l-azoniabicyclo[2.2.2]octane bromide (see WO 01/04118), 3(R)-l-phenethyl-3-(9H-xanthene-9-carbonyloxy)-l- azoniabicyclo[2.2.2]octane bromide or (3R)-3-[(2S)-2-cyclopentyl-2-hydroxy-2-thien-2- ylacetoxy]-l-(2-phenoxyethyl)-l-azoniabicyclo[2.2.2]actane bromide (see WO 01/04118); or a quaternary ammonium salt (such as [2-((S)-Cyclohexyl-hydroxy-phenyl-methyl)- oxazol-5-ylmethyl]-dimethyl-(3-phenoxy-propyl)-ammonium salt, [2-((R)-Cyclohexyl- hydroxy-phenyl-methyl)-oxazol-5 -ylmethyl] -dimethyl-(3 -phenoxy-propyl)-ammonium salt, [2-((R)-Cyclohexyl-hydroxy-phenyl-methyl)-oxazol-5 -ylmethyl]- dimethyl-(2- phenethyloxy-ethyl)-ammonium salt, [2-((R)-Cyclohexyl-hydroxy-phenyl-methyl)-oxazol- 5-ylmethyl]- [3-(3,4-dichloro-phenoxy)-propyl] dimethyl-ammonium salt, [2-((R)- Cyclohexyl-hydroxy-phenyl-methyl)-oxazol-5-ylmethyl]-[2-(3,4-dichloro-benzyloxy)- ethyl]- dimethyl-ammonium salt, [2-(4-Chloro-benzyloxy)-ethyl]-[2-((R)-Cyclohexyl- hydroxy-phenyl-methyl)-oxazol-5-ylmethyl]- dimethyl-ammonium salt, or (i?)-l-[2-(4- Fluoro-phenyl)-ethyl]-3-((5)-2-phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 -azonia- bicyclo[2.2.2]octane; wherein the counter-ion is, for example, chloride, bromide, sulfate, methanesulfonate, benzenesulfonate (besylate), toluenesulfonate (tosylate), napthalene- bissulfonate (napadisylate), phosphate, acetate, citrate, lactate, tartrate, mesylate, maleate, fumarate or succinate.

In one aspect of the invention a muscarinic antagonist is Aclidinium bromide, Glycopyrrolate (such as R,R-, R,S-, S,R-, or S,S-glycopyrronium bromide), Oxitropium bromide, Pirenzepine, telenzepine, Tiotropium bromide, 3(R)-(2-hydroxy-2,2-dithien-2- ylacetoxy)-l-(3-phenoxypropyl)-l-azoniabicyclo[2.2.2]octane bromide, or, 3(R)-I- phenethyl-3-(9H-xanthene-9-carbonyloxy)- 1 -azoniabicyclo[2.2.2]octane bromide, (3R)-3- [(2S)-2-cyclopentyl-2-hydroxy-2-thien-2-ylacetoxy]- 1 -(2-phenoxyethyl)- 1 - azoniabicyclo[2.2.2]actane bromide or Darotropium ((1R, 3r, 5S)-3-(2-cyano-2,2- diphenylethyl)-8,8-dimethyl-8-azoniabicyclo[3,2,l]octane bromide).

A p38 Inhibitor is, for example, a compound from WO 2005/042502, 681323, 856553, AMG548 (2-[[(2S)-2-amino-3-phenylpropyl]amino]-3-methyl-5-(2-naphthalenyl)- 6-(4-pyridinyl)-4(3H)-pyrimidinone), Array-797, AZD6703, Doramapimod, KC-706, PH 797804, R1503, SC-80036, SCIO469, 6-chloro-5-[[(2^5i?)-4-[(4-fluorophenyl)methyl]- 2,5-domethyl- 1 -piperazinyl]carbonyl]-JV,Λf, 1 -trimethyl-α-oxo- lH-indole-3-acetamide,

VX702 or VX745 (5-(2,6-dichlorophenyl)-2-(phenylthio)-6H-pyrimido[l,6-b]pyridazin-6- one).

A PPARγ agonist is, for example, Pioglitazone, Pioglitazone hydrochloride, Rosiglitazone Maleate, Rosiglitazone Maleate ((-)-enantiomer, free base), Rosiglitazone maleate/Metformin hydrochloride or Tesaglitizar.

A Protease Inhibitor is, for example, Alpha 1 -antitrypsin proteinase Inhibitor, EPI- HNE4, UT-77, ZD-0892 or a compound from WO 2006/004532, WO 2005/026123, WO 2002/0744767 or WO 22002/074751; or a TACE Inhibitor (for example DPC-333, Sch- 709156 or Doxy eye line).

A Statin is, for example, Atorvastatin, Lovastatin, Pravastatin, Rosuvastatin or Simvastatin.

A Thromboxane Antagonist is, for example, Ramatroban or Seratrodast.

A Vasodilator is, for example, A-306552, Ambrisentan, Avosentan, BMS-248360, BMS-346567, BMS-465149, BMS-509701, Bosentan, BSF-302146 (Ambrisentan),

Calcitonin Gene-related Peptide, Daglutril, Darusentan, Fandosentan potassium, Fasudil, Iloprost, KC- 12615 (Daglutril), KC- 12792 2AB (Daglutril) , Liposomal treprostinil, PS- 433540, Sitaxsentan sodium, Sodium Ferulate, TBC-11241 (Sitaxsentan), TBC-3214 (N- (2-acetyl-4,6-dimethylphenyl)-3-[[(4-chloro-3-methyl-5-isoxazolyl)amino]sulfonyl]-2- thiophenecarboxamide), TBC-3711, Trapidil, Treprostinil diethanolamine or Treprostinil sodium.

An ENAC (Epithelial Sodium-channel blocker) is, for example, Amiloride,

Benzamil, Triamterene, 552-02, PSA14984, PSA25569, PSA23682 or AER002. All the above second et seq active ingredients may be in the form of solvates, for example hydrates.

In one particular aspect the present invention provides a pharmaceutical product 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.

In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 6-fluoro-N-((ls,4s)-4-(6-fluoro-2,4- dioxo- 1 -(4'-(piperazin- 1 -ylmethyl)-biphenyl-3-yl)- 1 ,2-dihydropyrido[2,3-d]pyrimidin-

3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide or a salt thereof (such as the

(lS)-(+)-10-Camphorsulfonic acid or trihydrochloride salt), and a second active ingredient selected from: β₂ adrenoceptor agonist; a MABA compound; or, a muscarinic antagonist.

In yet another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 6-fluoro-N-((ls,4s)-4-(6- fluoro-2,4-dioxo-l-(4'-(piperazin-l-ylmethyl)-biphenyl-3-yl)-l,2-dihydropyrido[2,3- d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide or a salt thereof (such as the (lS)-(+)-10-Camphorsulfonic acid or trihydrochloride salt); a second active ingredient that is a β₂ adrenoceptor agonist; and, optionally, a third active ingredient that is selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; a CCRl antagonist; a chemokine antagonist (not CCRl); a corticosteroid; an IKK2 inhibitor; a muscarinic antagonist; or, a p38 inhibitor.

In yet another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 6-fluoro-N-((ls,4s)-4-(6- fluoro-2,4-dioxo-l-(4'-(piperazin-l-ylmethyl)-biphenyl-3-yl)-l,2-dihydropyrido[2,3- d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide or a salt thereof

(such as the (lS)-(+)-10-Camphorsulfbnic acid or trihydrochloride salt); a second active ingredient that is a MABA compound; and, optionally, a third active ingredient that is selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; a CCRl antagonist; a chemokine antagonist (not CCRl); a corticosteroid; an IKK2 inhibitor; or, a p38 inhibitor.

In a further aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 6-fluoro-N-((ls,4s)-4-(6- fluoro-2,4-dioxo-l-(4'-(piperazin-l-ylmethyl)-biphenyl-3-yl)-l,2-dihydropyrido[2,3- d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide or a salt thereof (such as the (lS)-(+)-10-Camphorsulfonic acid or trihydrochloride salt); a second active ingredient that is a muscarinic antagonist; and, optionally, a third active ingredient that is selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; a CCRl antagonist; a chemokine antagonist (not CCRl); a corticosteroid; an IKK2 inhibitor; or, a p38 inhibitor.

(lS)-(+)-10-Camphorsulfonic acid or trihydrochloride salt), and a second active ingredient is a β2-adrenoceptor agonist {for example: metaproterenol, isoproterenol, isoprenaline, albuterol, salbutamol (e.g. as sulphate), formoterol (e.g. as fumarate), salmeterol (e.g. as xinafoate), terbutaline, orciprenaline, bitolterol (e.g. as mesylate), pirbuterol, indacaterol, bambuterol (e.g. as hydrochloride), carmoterol (TA 2005, chemically identified as 2(1H)- Quinolone, 8-hydroxy-5-[ 1 -hydroxy-2-[[2-(4-methoxy-phenyl)- 1 -methylethyl]- amino]ethyl]-monohydrochloride, [R-(R*, R*)] also identified by Chemical Abstract Service Registry Number 137888-11-0 and disclosed in U.S. Patent No 4,579,854), a formanilide derivative (e.g. 3-(4-{[6-({(2R)-2-[3-(formylamino)-4-hydroxyphenyl]-2- hydroxyethyl}amino)hexyl]oxy}-butyl)-benzenesulfonamide as disclosed in WO 2002/76933), a benzenesulfonamide derivative (e.g. 3-(4-{[6-({(2R)-2-hydroxy-2-[4- hydroxy-3-(hydroxy-methyl)phenyl]ethyl} amino)-hexyl]oxy}butyl)benzenesulfonamide as disclosed in WO 2002/88167), an aryl aniline receptor agonist (for example as disclosed in WO 2003/042164 or WO 2005/025555), an indole derivative (for example as disclosed in WO 2004/032921 or US 2005/222144), a compound GSK 159797, GSK 159802, GSK 597901, GSK 642444 or GSK 678007; a N-[2-(Diethylamino)ethyl]-N-(2-{[2-(4-hydroxy- 2-oxo-2,3-dihydro-l,3-benzothiazol-7-yl)ethyl]amino}ethyl)-3-[2-(l- naphthyl)ethoxy]propanamide salt (for example Λ/-[2-(Diethylamino)ethyl]-iV-(2-{[2-(4- hydroxy-2-oxo-2,3-dihydro-l,3-benzothiazol-7-yl)ethyl]amino}ethyl)-3-[2-(l- naphthyl)ethoxy]propanamide dihydrobromide); a Λ/-[2-(Diethylamino)ethyl]-iV-(2-{[2-(4- hydroxy-2-oxo-2,3-dihydro- 1 ,3-benzothiazol-7-yl)ethyl]amino} ethyl)-3-[2-(3- chlorophenyl)ethoxy]propanamide salt (for example Λ/-[2-(Diethylamino)ethyl]-iV-(2- {[2- (4-hydroxy-2-oxo-2,3-dihydro- 1 ,3-benzothiazol-7-yl)ethyl]amino} ethyl)-3-[2-(3- chlorophenyl)ethoxy]propanamide dihydrobromide); a 7-[(li?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl] amino } propyl)thio]ethyl} amino)- 1 -hydroxy ethyl] -4-hydroxy- 1,3- benzothiazol-2(3H)-one salt (for example 7-[(li?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl] amino } propyl)thio]ethyl} amino)- 1 -hydroxy ethyl] -4-hydroxy- 1,3- benzothiazol-2(3H)-one dihydrobromide); or, a A/-Cyclohexyl-N³-[2-(3- fluorophenyl)ethyl]-Λ/-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3-benzothiazol-7- yl)ethyl] amino }ethyl)-β-alaninamide salt (for example A/-Cyclohexyl-N³-[2-(3- fluorophenyl)ethyl]-Λ/-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3-benzothiazol-7- yl)ethyl] amino }ethyl)-β-alaninamide di-D-mandelate salt)}.

In yet another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 6-fluoro-N-((ls,4s)-4-(6- fluoro-2,4-dioxo-l-(4'-(piperazin-l-ylmethyl)-biphenyl-3-yl)-l,2-dihydropyrido[2,3- d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide or a salt thereof (such as the (lS)-(+)-10-Camphorsulfonic acid or trihydrochloride salt), and a second active ingredient is a β2-adrenoceptor agonist {for example: metaproterenol, isoproterenol, isoprenaline, albuterol, salbutamol (e.g. as sulphate), formoterol (e.g. as fumarate), salmeterol (e.g. as xinafoate), terbutaline, orciprenaline, bitolterol (e.g. as mesylate), pirbuterol, indacaterol, bambuterol (e.g. as hydrochloride), a compound GSK 159797,

GSK 159802, GSK 597901, GSK 642444 or GSK 678007; a N-[2-(Diethylamino)ethyl]-N- (2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3-benzothiazol-7-yl)ethyl]amino}ethyl)-3-[2-(l- naphthyl)ethoxy]propanamide salt (for example Λ/-[2-(Diethylamino)ethyl]-iV-(2-{[2-(4- hydroxy-2-oxo-2,3-dihydro-l,3-benzothiazol-7-yl)ethyl]amino}ethyl)-3-[2-(l- naphthyl)ethoxy]propanamide dihydrobromide); a Λ/-[2-(Diethylamino)ethyl]-JV-(2-{[2-(4- hydroxy-2-oxo-2,3-dihydro- 1 ,3-benzothiazol-7-yl)ethyl]amino} ethyl)-3-[2-(3- chlorophenyl)ethoxy]propanamide salt (for example Λ/-[2-(Diethylamino)ethyl]-iV-(2-{[2- (4-hydroxy-2-oxo-2,3-dihydro- 1 ,3-benzothiazol-7-yl)ethyl]amino} ethyl)-3-[2-(3- chlorophenyl)ethoxy]propanamide dihydrobromide); a 7-[(li?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl] amino } propyl)thio]ethyl} amino)- 1 -hydroxy ethyl] -4-hydroxy- 1,3- benzothiazol-2(3H)-one salt (for example 7-[(li?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl] amino } propyl)thio]ethyl} amino)- 1 -hydroxy ethyl] -4-hydroxy- 1,3- benzothiazol-2(3H)-one dihydrobromide); or, a A/-Cyclohexyl-N³-[2-(3- fluorophenyl)ethyl]-Λ/-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3-benzothiazol-7- yl)ethyl] amino } ethyl)-β-alaninamide salt (for example JV-Cyclohexyl-iV³- [2-(3 - fluorophenyl)ethyl]-Λ/-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3-benzothiazol-7- yl)ethyl] amino }ethyl)-β-alaninamide di-D-mandelate salt)}.

In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 6-fluoro-N-((ls,4s)-4-(6-fluoro-2,4- dioxo- 1 -(4'-(piperazin- 1 -ylmethyl)-biphenyl-3-yl)- 1 ,2-dihydropyrido[2,3-d]pyrimidin- 3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide or a salt thereof (such as the (lS)-(+)-10-Camphorsulfonic acid or trihydrochloride salt), and a second active ingredient is a MABA compound having dual activity as both a muscarinic antagonist and as a β₂- adrenoceptor agonist, for example a MABA is a compound disclosed in: WO2004089892, WO2004106333, US20040167167, WO2005111004, WO2005051946, WO2006023457, WO2006023460, US20060223858, US20060223859, WO2007107828, WO2008000483, US7317102 or WO2008041095. For example the MABA is: biphenyl-2-ylcarbamic acid l-[2-(4-{[(R)-2-(3-formylamino-4-hydroxyphenyl)-2-hydroxyethylam-2,5- dimethylphenylcarbamoyl)ethyl]piperidin-4-yl ester; or succinic acid salt and 1 ,2- ethanedisulfonic of biphenyl-2-ylcarbamic acid l-[2-(2-chloro-4-{[(R)-2-hydroxy-2-(8- hydroxy-2-oxo-l,2-dihydroquinolin-5-yl)ethylmino]methyl}-5- methoxyphenyIcarbamoyl)ethyl]piperidin-4-yl ester; or naphthalene-l,5-disulfonic acid salt of biphenyl-2-ylcarbamic acid l-(9-[(R)-2-hydroxy-2-(8-hydroxy-2-oxo-l,2-dihydro- quinolin-5-yl)ethylamino]nonyl}piperidin-4-yl ester.

In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 6-fluoro-N-((ls,4s)-4-(6- fluoro-2,4-dioxo-l-(4'-(piperazin-l-ylmethyl)-biphenyl-3-yl)-l,2-dihydropyrido[2,3- d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide or a salt thereof (such as the (lS)-(+)-10-Camphorsulfonic acid or trihydrochloride salt), and a second active ingredient is a muscarinic antagonist, for example, Aclidinium bromide, Glycopyrrolate (such as R,R-, R,S-, S,R-, or S,S-glycopyrronium bromide), Oxitropium bromide, Pirenzepine, telenzepine, Tiotropium bromide, Darotropium ((1R, 3r, 5S)-3-(2- cyano-2,2-diphenylethyl)-8,8-dimethyl-8-azoniabicyclo[3,2,l]octane bromide), 3(R)-(2- hydroxy-2,2-dithien-2-ylacetoxy)- 1 -(3-phenoxypropyl)- 1 -azoniabicyclo[2.2.2]octane bromide (see WO 01/04118), or 3(R)-l-phenethyl-3-(9H-xanthene-9-carbonyloxy)-l- azoniabicyclo[2.2.2]octane bromide or (3R)-3-[(2S)-2-cyclopentyl-2-hydroxy-2-thien-2- ylacetoxy]-l-(2-phenoxyethyl)-l-azoniabicyclo[2.2.2]actane bromide (see WO 01/04118); or a quaternary ammonium salt (such as [2-((S)-Cyclohexyl-hydroxy-phenyl-methyl)- oxazol-5-ylmethyl]-dimethyl-(3-phenoxy-propyl)-ammonium salt, [2-((R)-Cyclohexyl- hydroxy-phenyl-methyl)-oxazol-5 -ylmethyl] -dimethyl-(3 -phenoxy-propyl)-ammonium salt, [2-((R)-Cyclohexyl-hydroxy-phenyl-methyl)-oxazol-5 -ylmethyl]- dimethyl-(2- phenethyloxy-ethyl)-ammonium salt, [2-((R)-Cyclohexyl-hydroxy-phenyl-methyl)-oxazol- 5-ylmethyl]- [3-(3,4-dichloro-phenoxy)-propyl] dimethyl-ammonium salt, [2-((R)- Cyclohexyl-hydroxy-phenyl-methyl)-oxazol-5-ylmethyl]-[2-(3,4-dichloro-benzyloxy)- ethyl]-dimethyl-ammonium salt, [2-(4-Chloro-benzyloxy)-ethyl]-[2-((R)-Cyclohexyl- hydroxy-phenyl-methyl)-oxazol-5 -ylmethyl] -dimethyl-ammonium salt, or (i?)-l-[2-(4- Fluoro-phenyl)-ethyl]-3-((5)-2-phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 -azonia- bicyclo[2.2.2]octane; wherein the counter-ion is, for example, chloride, bromide, sulfate, methanesulfonate, benzenesulfonate (besylate), toluenesulfonate (tosylate), napthalene- bissulfonate (napadisylate), phosphate, acetate, citrate, lactate, tartrate, mesylate, maleate, fumarate or succinate.

In a further aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 6-fluoro-N-((ls,4s)-4-(6- fluoro-2,4-dioxo-l-(4'-(piperazin-l-ylmethyl)-biphenyl-3-yl)-l,2-dihydropyrido[2,3- d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide or a salt thereof (such as the (lS)-(+)-10-Camphorsulfonic acid or trihydrochloride salt), and a second active ingredient is Oxitropium bromide, Tiotropium bromide, Aclidinium bromide, Glycopyrrolate (such as R,R-, R,S-, S,R-, or S,S-glycopyrronium bromide) or Darotropium ((1R, 3r, 5S)-3-(2-cyano-2,2-diphenylethyl)-8,8-dimethyl-8-azoniabicyclo[3,2,l]octane bromide).

In one aspect of the invention the muscarinic receptor antagonist is a long acting muscarinic receptor antagonist, that is a muscarinic receptor antagonist with activity that persists for more than 12 hours. Examples of long acting muscarinic receptor antagonists include tiotropium bromide.

In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 6-fluoro-N-((ls,4s)-4-(6-fluoro-2,4- dioxo- 1 -(4'-(piperazin- 1 -ylmethyl)-biphenyl-3-yl)- 1 ,2-dihydropyrido[2,3-d]pyrimidin- 3(4H)-yl)cyclohexyl)imidazo[ 1 ,2-a]pyridine-2-carboxamide or a salt thereof (such as the (lS)-(+)-10-Camphorsulfonic acid or trihydrochloride salt), and a second active ingredient is Tiotropium bromide.

The first active ingredient and the second active ingredient of the pharmaceutical product of the present invention may be administered simultaneously, sequentially or separately to treat respiratory diseases. By simultaneously is meant that the active ingredients are in admixture, or they could be in separate chambers of the same inhaler. By sequential it is meant that the active ingredients are administered, in any order, one immediately after the other. They still have the desired effect if they are administered separately, but when administered in this manner they are generally administered less than 4 hours apart, conveniently less than two hours apart, more conveniently less than 30 minutes apart and most conveniently less than 10 minutes apart, for example less than 10 minutes but not one immediately after the other.

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 be delivered to the lung and/or airways via oral administration in the form of a solution, suspension, aerosol or dry powder formulation. These dosage forms will usually include one or more pharmaceutically acceptable ingredients which may be selected, for example, from an adjuvant, carrier, binder, lubricant, diluent, stabilising agent, buffering agent, emulsifying agent, viscosity- regulating agent, surfactant, preservative, flavouring or colorant. 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 another embodiment the first and second active ingredients are administered via a single pharmaceutical composition (that is, the first and second active ingredients are in admixture). Therefore, the present invention further provides a pharmaceutical composition comprising, in admixture, a first active ingredient which is 6-fluoro-N- ((I s,4s)-4-(6-fluoro-2,4-dioxo- 1 -(4'-(piperazin- 1 -ylmethyl)-biphenyl-3-yl)- 1 ,2- dihydropyrido[2,3-d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2- carboxamide or a salt thereof (such as the (lS)-(+)-10-Camphorsulfonic acid or trihydrochloride salt), and a second active ingredient as defined above. The pharmaceutical composition optionally further comprises a pharmaceutically acceptable adjuvant, diluent or carrier.

The pharmaceutical compositions of the present invention can be prepared by mixing the 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 the first and second active ingredients 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 first active ingredient is administered via inhalation. When administered via inhalation the dose of the first active ingredient (that is 6-fluoro-N-((ls,4s)-4-(6-fluoro-2,4-dioxo-l-(4'-(piperazin-l-ylmethyl)-biphenyl-3-yl)- l,2-dihydropyrido[2,3-d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2- carboxamide in: salt form, solvate form, or, solvate of salt form) 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 is administered by inhalation. When administered via inhalation the dose of the second active ingredient 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 another embodiment the present invention provides a pharmaceutical product wherein the molar ratio of first active ingredient to second active ingredient is from 1 : 1000 to 1000:1, such as from 1 :100 to 100:1, for example from 1 :50 to 50:1, for example 1 :20 to 20:1.

In one embodiment, the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient as defined above, and a second active ingredient as defined above, wherein each active ingredient is formulated for inhaled administration. In a further aspect of this embodiment, the pharmaceutical product is in the form of a pharmaceutical composition comprising the first and second active ingredients in admixture, and which composition is formulated for inhaled administration. The active ingredients of the present invention are conveniently delivered via oral administration by inhalation to the lung and/or airways in the form of a solution, suspension, aerosol or dry powder (such as an agglomerated or ordered mixture) formulation. For example a metered dose inhaler device may be used to administer the active ingredients, dispersed in a suitable propellant and with or without an additional excipient such as ethanol, a surfactant, lubricant or stabilising agent. A suitable propellant includes a hydrocarbon, chlorofluorocarbon or a hydrofluoroalkane (e.g. heptafluoroalkane) propellant, or a mixture of any such propellants, for example in a pressurised metered dose inhaler (pMDI). Preferred propellants are P 134a and P227, each of which may be used alone or in combination with other another propellant and/or surfactant and/or other excipient. A nebulised aqueous suspension or, preferably, solution may also be employed, with or without a suitable pH and/or tonicity adjustment, either as a unit-dose or multi-dose formulation. A suitable device for delivering a dry powder is Turbuhaler®.

The pharmaceutical product of the present invention can, for example, be administered: via an inhaler having the first and second active ingredients in separate chambers of the inhaler such that on administration the active ingredients mix in either the mouthpiece of the inhaler or the mouth of a patient or both (for simultaneous use); or, where the first and second active ingredients are in separate inhalers, via separate inhalers (for separate or sequential use); or the first and second active ingredients are in admixture in an inhaler when the inhaler is supplied to a patient (for simultaneous use).

A dry powder inhaler may be used to administer the active ingredients, alone or in combination with a pharmaceutically acceptable carrier (such as lactose), 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 is available. The combination of the present invention may be used to treat diseases of the respiratory tract such as obstructive diseases of the airways including: asthma, including bronchial, allergic, intrinsic, extrinsic, exercise-induced, drug-induced (including aspirin and NS AID-induced) and dust-induced asthma, both intermittent and persistent and of all severities, and other causes of airway hyper-responsiveness; chronic obstructive pulmonary disease (COPD); bronchitis, including infectious and eosinophilic bronchitis; emphysema; bronchiectasis; cystic fibrosis; sarcoidosis; farmer's lung and related diseases; hypersensitivity pneumonitis; lung fibrosis, including cryptogenic fibrosing alveolitis, idiopathic interstitial pneumonias, fibrosis complicating anti-neoplastic therapy and chronic infection, including tuberculosis and aspergillosis and other fungal infections; complications of lung transplantation; vasculitic and thrombotic disorders of the lung vasculature, and pulmonary hypertension; antitussive activity including treatment of chronic cough associated with inflammatory and secretory conditions of the airways, and iatrogenic cough; acute and chronic rhinitis including rhinitis medicamentosa, and vasomotor rhinitis; perennial and seasonal allergic rhinitis including rhinitis nervosa (hay fever); nasal polyposis; acute viral infection including the common cold, and infection due to respiratory syncytial virus, influenza, coronavirus (including SARS) and adenovirus.

Accordingly, the present invention further provides a pharmaceutical product according to the invention for simultaneous, sequential or separate use in therapy. The present invention further provides the use of a pharmaceutical product according to the invention in the manufacture of a medicament for the treatment of a respiratory disease, in particular chronic obstructive pulmonary disease, asthma, rhinitis, emphysema or bronchitis (such as chronic obstructive pulmonary disease or asthma; for example chronic obstructive pulmonary disease). 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 as defined above; and,

(b) a therapeutically effective dose of a second active ingredient as defined above; to a patient in need thereof. In a further aspect the present invention provides the use of a pharmaceutical product, kit or composition as hereinbefore described for the treatment of a respiratory disease, in particular chronic obstructive pulmonary disease, asthma, rhinitis, emphysema or bronchitis (such as chronic obstructive pulmonary disease or asthma; for example chronic obstructive pulmonary disease). 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.

Scheme 89 of WO 2007/054831 discloses a two stage method for making a boronic acid derivative of JV-benzyl piperazine. There is also presented the invention of a 'one-pot' process for the preparation of a compound of formula (I):

wherein R* is one or more of the same or different substituents on the phenyl ring which do not react under reductive conditions; and R¹ and R² are the same or different substituents that can optionally form a substituted ring; the process comprising reacting a compound of formula (II):

with a compound of formula (III):

-R¹

HN (IN)

-R' under reductive conditions, (for example in the presence of an alkali metal triacetoxyborohydride and a suitable solvent (such as tetrahydrofuran, 2- methyltetrahydrofuran, 1 ,2-dichloromethane, JV-methylpyrollidinone or acetonitrile; and in particular tetrahydrofuran or 2-methyltetrahydrofuran) at ambient temperature (for example in the range 5-25⁰C).

In one aspect of the invention R* is, for example, hydrogen.

In another aspect of the invention R¹ and R² are, independently, hydrogen, Ci_₆ alkyl or C2-6 alkyl (substituted by halo or Ci_4 alkoxy); or R¹ and R² join to form a ring {such as a pyrrolidinyl, piperidinyl, piperazinyl (protected on the second ring nitrogen, for example by a BOC group), morpholinyl, homopiperidinyl or homomorpholinyl ring} . In yet another aspect R¹ and R² join to form a piperazinyl ring protected on the second ring nitrogen by a BOC group.

In a further aspect of the invention the boronic acid group of the compound of formula (I) is para to the CH₂N(CH₂R¹XCH₂R²) group.

In another aspect the present invention provides an intermediate compound of formula (IV):

In a further aspect the present invention provides the intermediate compounds: 5-Fluoro-2-(3-iodophenylamino)nicotinic acid; tert-Butyl cis-4-(5 -fluoro-2-(3 -iodophenylamino)nicotinamido)cyclohexylcarbamate; fert-Butyl cis-4-(6-fluoro-l-(3-iodophenyl)-2,4-dioxo-l,2-dihydropyrido[2,3-d]pyrimidin- 3(4H)-yl) cyclohexylcarbamate;

3-(cis-4-Aminocyclohexyl)-6-fluoro- 1 -(3-iodophenyl) pyrido[2,3-d]pyrimidine- 2,4(1 H,3H)-dione; 2-Ethoxycarbonyl-6-fluoro-2-hydroxy-2,3-dihydro-lH-imidazo[l,2-a]pyridin-4-ylium- bromide; or,

6-Fluoro-N-(cis-4-(6-fluoro- 1 -(3-iodophenyl)-2,4-dioxo- 1 ,2-dihydropyrido[2,3- d]pyrimidin-3(4H)-yl) cyclohexyl)imidazo[ 1 ,2-a]pyridine-2-carboxamide hydrochloride.

General Preparative Methods

There follow methods for the preparation of certain compounds recited above.

¹H NMR spectra were recorded on a Varian Inova 400 MHz or a Varian Mercury-VX 300 MHz instrument. The central peaks of chloroform-<i (5_H 7.27 ppm),

(5_H 2.50 ppm), acetonitrile-^ (5_H 1.95 ppm) or methanol-^ (5_H 3.31 ppm) were used as internal references. Column chromatography was carried out using silica gel (0.040-0.063 mm, Merck). Unless stated otherwise, starting materials were commercially available. All solvents and commercial reagents were of laboratory grade and were used as received.

The following method was used for LC/MS analysis:

Instrument Agilent 1100; Column Waters Symmetry 2.1 x 30 mm; Mass APCI; Flow rate 0.7 ml/min; Wavelength 254 nm; Solvent A: water + 0.1% TFA; Solvent B: acetonitrile + 0.1% TFA ; Gradient 15-95%/B 8 min, 95% B 1 min. Analytical chromatography was run on a Symmetry Cis-column, 2.1 x 30 mm with 3.5 μm particle size, with acetonitrile/water/0.1% trifluoroacetic acid as mobile phase in a gradient from 5% to 95% acetonitrile over 8 minutes at a flow of 0.7 ml/min.

The abbreviations or terms used in the preparations have the following meanings:

SCX: Solid phase extraction with a sulfonic acid sorbent

HPLC: High performance liquid chromatography

DMF: Λ/,Λ/-Dimethylformamide

THF: Tetrahydrofuran

NMP: N-Methylpyrrolidinone

HATU: O-(7-Azabenzotriazol- 1 -yl)-ΛWΛT,ΛP-tetramethyluroniurn hexafluorophosphate

TFA: Trifluoroacetic acid

Preparation 1

7V-Cyclohexyl-7V³-[2-(3-fluorophenyl)ethyl]-7V-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro- l,3-benzothiazol-7-yl)ethyl] amino} ethyl)-β-alaninamide bis-trifluoroacetic acid salt

i) Benzyl (2,2-dimethoxyethyl)[2-(4-hydroxy-2-oxo-2,3-dihydro- 1 ,3-benzothiazol-7- yl)ethyl] carbamate

7-(2-Aminoethyl)-4-hydroxy-l,3-benzothiazol-2(3H)-one hydrobromide (20 g) was dissolved in a mixture of THF (300 ml) and water (150 ml). Sodium hydrogen carbonate (5.77 g) was added and the mixture stirred for 15 min. Acetic acid (7.86 ml) was added, followed by dimethoxyacetaldehyde (14.9 g, 12.91 ml) and the mixture stirred for a further 30 min. Sodium cyanoborohydride (8.64 g) was added portion-wise over 10 min and the solution stirred for a further 20 h. Ethyl acetate (500 ml) and a solution of sodium hydrogen carbonate (17.33 g) in water (250 ml) were added, the mixture was stirred vigorously, benzyl chloroformate (8.78 g, 7.35 ml) was added, and the mixture stirred for 2 h. The organic layer was separated, washed with water, 0.1M aq. HCl, water and brine, dried (anhydrous Na₂SO₄), filtered and evaporated. The resulting material was purified by flash chromatography on silica using 10% methanol in dichloromethane as eluent to give the sub-title compound as a light brown gum (23.1 g).

¹H NMR δ(DM_Sθ) H.60 (IH, s), 9.90 (IH, s), 7.39-7.12 (5H, m), 6.73 (2H, m), 5.05 (2H, m), 4.43 (0.5H, t), 4.35 (0.5H, t), 3.41 (2H, m), 3.33 (1.5H, s), 3.27 (3H, s), 3.22 (1.5H, s), 3.19 (2H, m), 2.69 (2H, q).

MS (APCI+) 433 [M+H]⁺

ii) Benzyl[2-(4-hydroxy-2-oxo-2,3-dihydro-l ,3-benzothiazol-7-yl)ethyl](2- oxoethyl)carbamate

The acetal from step i) (5 g) was dissolved in acetone (100 ml), 2M HCl in dioxane (50 ml) was added and the mixture stirred for 3 h. Concentrated HCl (2 ml) was added and mixture stirred for a further 20 h. Toluene (100 ml) was added and the solvents removed in vacuo. The residue was dissolved in THF (200 ml), toluene (100 ml) added, and the solvents removed in vacuo (x2) to give the sub-title compound as an off white solid (4.5 g). ¹H NMR δ (DM_SO) 11.61 (IH, m), 9.91 (IH, m), 9.41 (IH, s), 7.31 (5H, m), 6.74 (2H, m), 5.01 (2H, m), 4.04 (2H, d), 3.46 (2H, t), 2.69 (2H, t).

MS (APCI +) 387 [M+H]⁺

iii) Benzyl [2-(cyclohexylamino)ethyl][2-(4-hydroxy-2-oxo-2,3-dihydro-l,3-benzothiazol- 7-yl)ethyl] carbamate

The product of step iii) (0.5 g) was added to a solution of cyclohexylamine (0.23 g, 0.33 ml) in a mixture of THF (10 ml) and water (1 ml) and the mixture was stirred for 15 min. Sodium cyanoborohydride (0.17 g) was added, followed by acetic acid (0.24 g, 0.23 ml) and the reaction stirred for a further 2 h. The reaction was quenched with saturated aqueous sodium hydrogen carbonate, extracted with ethyl acetate, washed with brine, dried (anhydrous Na₂SO₄), filtered and evaporated to give the sub-title compound as a light brown gum (0.6 g).

¹H NMR 90⁰C δ (DM_SO) 7.40-7.50 (m, 5H), 6.86 (d, IH), 6.80 (d, IH), 5.18 (s, 2H), 3.72 (t, 2H), 3.56 (t, 2H), 2.94 (t, 2H), 2.83 (t, 2H), 1.96 (m, 2H), 1.84 (m, 4H), 1.68 (m, IH), 1.29 (m, 4H).

MS (APCI +) 470 [M+H]⁴

iv) Benzyl {2-[acryloyl(cyclohexyl)amino]ethyl}[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3- benzothiazol-7-yl)ethyl]carbamate

The amine as prepared in step iii) (1.57 g) was dissolved in dichloromethane (20 ml), chlorotrimethylsilane (1.29 ml) and triethylamine (1.91 ml) were added, and the mixture was stirred at room temperature for 1 h. The mixture was cooled to O⁰C, acryloyl chloride (336 ul) added, and the mixture was stirred, warming to room temperature, for 3 h. The reaction mixture was diluted with dichloromethane, washed with saturated sodium hydrogen carbonate, then with water, dried (anhydrous Na₂SO₄), filtered and evaporated. The residue was purified by flash chromatography on silica using ethyl acetate (30, 50, 70, 100%) in isohexane as eluent to give the sub-title compound (l.lg).

MS (APCI +) 524 [M+H]⁺

v) N-Cyclohexyl-Λ/³-[2-(3-fluorophenyl)ethyl]-N-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro- l,3-benzothiazol-7-yl)ethyl]amino}ethyl)-β-alaninamide bis-trifluoroacetic acid salt

The acrylamide as prepared in step iv) (1 ml of a 0.33M solution in ethanol) was treated with 3-fluorophenethylamine (97 ul) and the mixture was stirred at 5O⁰C for 18 h. The product was purified by SCX chromatography eluting with IN ammonia in methanol. The solvents were removed in vacuo and the residue was re-dissolved in dichloromethane (0.5 ml). This solution was cooled in an ice/water bath, hydrogen bromide 30 wt % solution in acetic acid (0.5 ml) was added, and the mixture was stirred at room temperature for 2 h. Toluene (1 ml) was added to the reaction and all solvents were removed in vacuo. The residue was azeotroped with toluene, then ethanol (x2) before being purified by reverse phase HPLC (5-40% acetonitrile in aqueous TFA). The residue was triturated with diethyl ether to give the title compound as a white solid (30 mg).

¹H NMR δ (DMSθ) H.73 (IH, s), 10.13 (IH, s), 8.84-8.48 (4H, m), 7.43-7.34 (IH, m), 7.18- 7.08 (3H, m), 6.86 (IH, d), 6.75 (IH, d), 3.59-3.45 (4H, m), 3.30-3.10 (5H, m), 3.03-2.93 (4H, m), 2.85-2.77 (4H, m), 1.81-1.03 (1OH, m).

MS (Multimode +) 529 [(M-salt)+H]⁺

Preparation 2

7V-Cyclohexyl-7V³-[2-(3-fluorophenyl)ethyl]-7V-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro- l,3-benzothiazol-7-yl)ethyl] amino} ethyl)-β-alaninamide dihydrobromide salt

i) tert-Butyl Λ/-[2-(3-fluorophenyl)ethyl]-β-alaninate

tert-Butyi acrylate (5.61 ml) was added to a solution of 3-fluorophenethylamine (5.0 ml) in ethanol (200 ml) and the mixture stirred at room temperature for 2 days. The solvent was removed in vacuo to afford the sub-title compound as an oil (9.6 g).

¹H NMR δ ₍cD_Ci3) δ 7.28 - 7.20 (m, IH), 7.01 - 6.85 (m, 3H), 2.88 - 2.74 (m, 6H), 2.41 (t, J = 6.5 Hz, 2H), 1.42 (s, 9H).

MS (APCI ⁺) 268 [M+H]⁺

ii) fert-Butyl Λ/-[(benzyloxy)carbonyl]-Λ/-[2-(3-fluorophenyl)ethyl]-β-alaninate

Benzyl chloro formate (5.57 ml) was added dropwise over 5 minutes to a solution of tert- butyl Λ/-[2-(3-fluorophenyl)ethyl]-β-alaninate, as prepared in step i) (9.5 g) and triethylamine (5.94 ml) in dichloromethane (100 ml) at ~ 5 ⁰C. The reaction was allowed to attain room temperature and stirred overnight. The solvent was removed in vacuo and the residue purified by flash chromatography on silica using 10 % ethyl acetate in isohexane as eluent to give the sub-title compound as an oil (11.5 g).

IH NMR (DMSO) δ 7.37 - 7.22 (m, 5H), 7.06 - 6.91 (m, 3H), 5.05 (s, 2H), 3.46 (t, J = 7.3 Hz, 2H), 3.39 (t, J = 7.0 Hz, 2H), 2.81 (t, J = 7.4 Hz, 2H), 2.41 (t, J = 7.2 Hz, 2H), 1.38 (s, 9H).

MS (APCI ⁺) 402 [M+H]⁺

iii) Λ/-[(Benzyloxy)carbonyl]-Λ/-[2-(3-fluorophenyl)ethyl]-β-alanine

Trifluoroacetic acid (50 ml) was added to a solution of the tert-hvXy\ ester, as prepared in step ii) (11.5 g) in dichloromethane (50 ml) and the mixture was stirred at room temperature for 2 h. The solvent was removed in vacuo and the oil azeotroped with toluene (x2) to afford the sub-title compound as a viscous oil (10.5 g).

¹H NMR δ (DM_SO) δ 7.41 - 7.27 (m, 6H), 7.08 - 6.94 (m, 3H), 5.04 (d, J = 25.9 Hz, 2H), 3.45 (t, J = 7.4 Hz, 2H), 3.38 (s, 2H), 2.84 - 2.76 (m, 2H), 2.45 (t, J = 7.0 Hz, 2H). MS (APCI ^") 344 [M-H]^"

iv) Benzyl {3-[cyclohexyl(2,2-dimethoxyethyl)amino]-3-oxopropyl} [2-(3- fluorophenyl)ethyl]carbamate

To a solution of Λ/-[(benzyloxy)carbonyl]-Λ/-[2-(3-fluorophenyl)ethyl]-β-alanine, as prepared in step iii) (5g) dissolved in dichloromethane (50 ml) with stirring under nitrogen was added dimethylformamide (2 drops) followed by oxalyl choride (1.64 ml) dropwise over 10 min. The mixture was stirred at room temperature for 1 h, concentrated in vacuo and redissolved in dichloromethane (25 ml). The solution was added dropwise to a preformed mixture of N-(2,2-dimethoxyethyl)cyclohexanamine (2.71 g) and triethylamine (3.0 ml) in dichloromethane (25 ml) at O⁰C under nitrogen. The mixture was stirred at O⁰C for 1 h, then water (25 ml) was added and the layers were separated. The organic layer was washed with 2M hydrochloric acid, saturated aqueous sodium bicarbonate and brine before being dried (anhydrous MgSO₄) filtered and concentrated in vacuo to give the subtitle compound as an oil (7.45 g).

¹H NMR δ (DM_Sθ) 7.35 (5H, s), 7.25-7.15 (IH, m), 7.02-6.76 (3H, m), 5.12 (2H, d), 4.62- 4.52 (IH, m), 4.39-4.26 (0.5H, m), 4.23-4.09 (0.5H, m), 3.59-3.46 (4H, m), 3.38 (6H, s), 3.35-3.23 (2H, m), 2.92-2.45 (4H, m), 1.88-0.99 (1OH, m)

MS: APCI (+ve): 515 [M+H]⁺

v) Benzyl {3-[cyclohexyl(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3-benzothiazol-7- yl)ethyl] amino } ethyl)amino] -3 -oxopropyl} [2-(3 -fluorophenyl)ethyl]carbamate

/?αra-Toluenesulfonic acid monohydrate (10.4 g) was added to a solution of the product from step i) (9.4g) in dichloromethane (94 ml). The mixture was stirred at room temperature for 40 min and a solution of saturated aqueous sodium bicarbonate (4.6 g) in water (100 ml) was added. The layers were separated and the organic phase was washed with saturated aqueous sodium bicarbonate (50 ml) and water (50 ml) before being dried (anhydrous MgSO₄), filtered and concentrated. The resulting oil was redissolved in N- methylpyrrolidinone (30 ml) and added to a solution of 7-(2-aminoethyl)-4-hydroxy-l,3- benzothiazol-2(3H)-one hydrobromide (6.0 g) and triethylamine (2.9 ml) in N- methylpyrrolidinone (30 ml) and water (3 ml). Sodium triacetoxyborohydride (6.0 g) was added and the mixture was stirred at room temperature for 3 h before being poured into water (600 ml) and extracted with ethyl acetate (2 x 150 ml). The organic layer was washed with aqueous sodium chloride (100 ml). A solid precipitated from the organic layer, which was partially concentrated in vacuo, and the precipitate was collected by filtration and washed with ethyl acetate to give the sub-title compound as a colourless solid (7-7 g).

¹H NMR δ _(DMSO) δ 7.41-7.24 (5H, m), 7.10-6.93 (3H, m), 6.86 (IH, d), 6.77 (IH, m), 5.05 (2H, d), 3.63-3.26 (8H, m), 3.13-3.01 (2H, m), 2.99-2.76 (6H, m), 2.62-2.52 (IH, m), 1.79- 0.95 (1OH, m)

MS: APCI (+ve): 663 [M+H]⁺

vi) N-Cyclohexyl-Λ/³-[2-(3-fluorophenyl)ethyl]-N-(2- {[2-(4-hydroxy-2-oxo-2,3-dihydro- 1 ,3-benzothiazol-7-yl)ethyl]amino} ethyl)-β-alaninamide dihydrobromide salt

To a solution of the product from step v) (1 g) in acetic acid (3 ml) stirred at room temperature was added hydrobromic acid in acetic acid (33%, 3 ml). The mixture was stirred for 80 min and then tert-butyl methyl ether (8 ml) was added. The mixture was stirred for 5 min and then filtered, washing with tert-butyl methyl ether (8 ml).

Purification by recrystallisation from hot ethanol (20 ml) gave the title compound (0.82 g) as a solid.

¹H NMR δ (DM_SO) 11-72 (IH, s), 10.08 (IH, s), 8.60 (4H, s), 7.39 (IH, q), 7.22-7.03 (3H, m), 6.88 (IH, d), 6.81-6.72 (IH, m), 3.65-3.47 (3H, m), 3.32-3.08 (6H, m), 3.07-2.95 (4H, m), 2.94-2.81 (4H, m), 1.76 (3H, t), 1.68-1.22 (5H, m), 1.19-1.02 (2H, m)

MS: APCI (+ve): 529 [M+H]⁺

Preparation 3

7V-Cyclohexyl-7V³-[2-(3-fluorophenyl)ethyl]-7V-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro- l,3-benzothiazol-7-yl)ethyl] amino} ethyl)-β-alaninamide di-D-Mandelate salt

A portion of the dihydrobromide from Preparation 2 was suspended in tetrahydrofuran and water (5:1), treated with a solution of saturated aqueous sodium bicarbonate (3 mol eq) in water, and the mixture stirred for 15 min. The tetrahydrofuran was removed in vacuo, sodium chloride was added, and the mixture was extracted with chloroform. The combined organic fractions were washed with water and brine, dried (anhydrous Na₂SO₄), filtered and treated with a solution of D-mandelic acid (3 mol eq) in acetonitrile (40 ml). The mixture was stirred for 2 h, filtered, washed with acetonitrile and dried to give the title compound as a colourless solid.

¹H NMR δ (DM_SO) 7.40 (4H, d), 7.38-7.14 (7H, m), 7.05 (3H, t), 6.82-6.67 (2H, m), 4.75- 4.69 (2H, m), 4.10-3.97 (0.5H, m), 3.53-3.44 (0.5H, m), 3.35-3.22 (2H, m), 3.07-2.97 (4H, m L), 2.92-2.73 (6H, m), 2.72-2.61 (4H, m), 1.78-1.69 (2H, m), 1.65-1.55 (2H, m), 1.52-1.17 (5H, m), 1.13-1.00 (IH, m)

MS: APCI (+ve): 529 [M+H]⁺ Preparation 4

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

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

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

Chiral HPLC 80:20 ώohexane : ethanol (isocratic). Chiracel OJ-H 4.6mm x 50mm Retention time 1.09min.

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

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

Chiral HPLC 80:20 ώohexane : ethanol (isocratic). Chiracel OJ-H 4.6mm x 50mm Retention time 2.52min. 1H NMR (400 MHz, CDCl₃) δ 7.56 - 7.49 (2H, m), 7.35 - 7.20 (3H, m), 3.68 (3H, s), 2.54 - 2.45 (2H, m), 2.41 - 2.32 (2H, m), 1.64 - 1.54 (7H, m), 1.50 - 1.42 (2H, m).

Preparation 4 (Form A): (R)-l-[2-(4-Fluoro-phenyl)-ethyl]-3-((S)-2-phenyl-2- piperidin-l-yl-propionyloxy)-l-azonia-bicyclo [2.2.2] octane bromide (Form A) Intermediate B: 2-Phenyl-2-piperidin-l-yl-propionic acid (i?)-(l-aza-bicyclo[2.2.2]oct-3- yl) ester (Isomer 1)

A mixture of 2-phenyl-2-piperidin-l-yl-propionic acid methyl ester (Intermediate A, Isomer 1) (0.9 g), (i?)-quinuclidin-3-ol (1.157 g) and sodium hydride (60% in mineral oil, 0.335 g) in dry toluene (20 mL) was heated at 120⁰C under an atmosphere of nitrogen for 8h. The cooled reaction mixture was diluted with water (100 mL) and extracted with diethyl ether (2 x 150 mL). The combined extracts were dried (MgSO₄) and concentrated to give an oil. The crude product was purified by flash column chromatography on silica eluting with (ethyl acetate / methanol 9:1) to afford the titled compound (0.500 g).

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

(R)-l-[2-(4-Fluoro-phenyl)-ethyl]-3-((S)-2-phenyl-2-piperidin-l-yl-propionyloxy)-l- azonia-bicyclo [2.2.2] octane bromide (Form A)

2-Phenyl-2-piperidin-l-yl-propionic acid (7?)-(l-aza-bicyclo[2.2.2]oct-3-yl) ester (Intermediate B, Isomer 1) (3 g) in acetonitrile (25 mL) was treated with l-(2-bromoethyl)- 4-fluorobenzene (2.384 g) and the mixture stirred at RT for 24 h. The mixture was concentrated to dryness, and the residue purified on silica gel eluting with 10% methanol in dichloromethane. The product containing fractions were combined, concentrated to dryness and the foam residue re-dissolved in acetonitrile (20 mL). To the solution was added diethyl ether (40 mL) and the resulting solid collected by filtration. The solid was dissolved in hot acetone (75 mL) and then allowed to cool overnight. The resulting solid was collected by filration and dried at 50⁰C to afford the titled compound (3.70 g).

m/e 465 [M]⁺

¹H NMR (400 MHz, DMSO) δ 7.58 - 7.54 (2H, m), 7.40 - 7.32 (4H, m), 7.31 - 7.26 (IH, m), 7.23 - 7.16 (2H, m), 5.14 - 5.09 (IH, m), 3.95 - 3.85 (IH, m), 3.62 - 3.51 (IH, m), 3.50 - 3.36 (4H, m), 3.25 - 3.16 (2H, m), 2.95 (2H, t), 2.48 - 2.31 (4H, m), 2.24 - 2.18 (IH, m), 2.02 - 1.69 (4H, m), 1.57 (3H, s), 1.56 - 1.48 (4H, m), 1.47 - 1.40 (2H, m).

Single crystal X-ray diffraction data obtained for Preparation 1 proved the structure to be (R)- 1 -[2-(4-Fluoro-phenyl)-ethyl]-3-((5)-2-phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 - azonia-bicyclo[2.2.2]octane bromide. The data set was collected at RT with graphite monochromatized MoK(a) radiation on a KappaCCD Single-Crystal X-Ray diffractometer equipped with an k-axis goniometer and a CCD area detector (Nonius, 1998). The diffraction raw data were processed within the Denzo-SMN program package (Otwinowski & Minor, 1998) converting the information from the digital image frame to a file containing h, k, 1 indices, background and Lp corrected intensities of the diffraction spots, along with estimate of errors.

On the basis of the crystal structure determined for Preparation 4, the absolute configuration of Intermediate A - Isomer 1 used has been assigned as (5)-2-Phenyl-2- piperidin- 1 -yl-propionic acid methyl ester.

Analysis of Preparation 4 Form A: (R)-l-[2-(4-Fluoro-phenyl)-ethyl]-3-((S)-2-phenyl- 2-piperidin-l-yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide (Form A) A sample of crystalline Preparation 4 bromide Form A obtained by the procedure described above was analysed by XRPD (PANalytical X'Pert or Cubix system), GVS, DSC and TGA.

The melting temperature of Preparation 4 bromide Form A as determined by DSC gave found a double endothermic events occurring at 171°C (1^st onset) and 183 ⁰C (2^nd onset) (±2°C). Weight loss observed prior to melting by TGA was negligible. GVS determination gave 0.1% weight increase (%w/w) at 80% RH (±0.2%).

An XRPD spectrum of Preparation 4 bromide Form A is presented in Figure 1.

Preparation 4 Form C: (R)-l-[2-(4-Fluoro-phenyl)-ethyl]-3-((S)-2-phenyl-2- piperidin-l-yl-propionyloxy)-l-azonia-bicyclo [2.2.2] octane bromide (Form C)

(R)- 1 -[2-(4-Fluoro-phenyl)-ethyl]-3-((5)-2-phenyl-2-piperidin- 1 -yl-propionyloxy)- 1 - azonia-bicyclo[2.2.2]octane bromide (above) (1 g) was dissolved in methanol (5 mL) and the mixture warmed to 60⁰C. The mixture was allowed to cool to 40⁰C whereupon solid started to form and the mixture was then re -heated to 50⁰C. Three 10 mL aliquots of methyl acetate were added to the mixture which was then allowed to slowly cool to room temperature and stirred for 18 h. The resulting solid was collected by filtration and then dried under reduced pressure at 50⁰C to afford the titled compound (50 mg).

¹H NMR (400 MHz, DMSO) δ 7.51 - 7.60 (2H, m), 7.31 - 7.41 (4H, m), 7.25 - 7.31 (IH, m), 7.13 - 7.21 (2H, m), 5.08 - 5.15 (IH, m), 3.88 - 3.97 (IH, m), 3.53 - 3.63 (IH, m), 3.38 - 3.52 (4H, m), 3.15 - 3.26 (2H, m), 2.92 - 3.01 (2H, m), 2.31 - 2.48 (4H, m), 2.20 - 2.25 (IH, m), 1.72 - 2.04 (4H, m), 1.58 (3H, s), 1.48 - 1.56 (4H, m), 1.39 - 1.48 (2H, m).

Analysis of Preparation 4 Form C: (if)-l-[2-(4-Fluoro-phenyl)-ethyl]-3-((S)-2-phenyl- 2-piperidin-l-yl-propionyloxy)-l-azonia-bicyclo[2.2.2]octane bromide (Form C)

A sample of crystalline Preparation 4 Crystalline Form C obtained by the procedure described above was analysed by XRPD (PANalytical X'Pert or Cubix system), GVS, DSC and TGA. The melting temperature of Preparation 4 bromide Form C as determined by DSC was found to be 184°C (onset) (±2°C). Weight loss observed prior to melting by TGA was 4%. GVS determination gave 4% weight increase (%w/w) at 80% RH (±0.2%).

An XRPD spectrum of Preparation 4 bromide Form C is presented in Figure 2.

Preparation 5

Preparation of 6-fluoro-N-(( 1 s,4s)-4-(6-fluoro-2,4-dioxo- 1 -(4'-(piperazin- 1 -ylmethyl)- biphenyl-3-yl)- 1 ,2-dihydropyrido[2,3-d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[ 1 ,2- a]pyridine-2-carboxamide and its (lS)-(+)-10-Camphorsulfonic acid salt)

Stage I: Preparation of 4-((4-(tert-butoxycarbonyl)piperazin-l-yl)methyl)phenylboronic acid (3)

M W 320 20

MF C₁₆H₂₅BN₂O₄

Charged 4-formylphenylboronic acid (1) (41.14 g, 40.00 g @100%, 0.267 mol, 1.00 mol eq) into a 1.0 L four-necked RB flask, equipped with a mechanical stirrer, 250.0 mL pressure equalising funnel, thermo-well and a nitrogen inlet at 20-25⁰C. Charged tetrahydrofuran (120.0 mL, 3.00 rel vol) and stirred the mass for 5-10 min. In another 500.0 mL four-necked RB flask, equipped with a magnetic stirrer, thermo well and nitrogen inlet, a solution of tert-butyl piperazine-1-carboxylate (2) (66.12 g, 64.60 g

@100%, 0.347 mol, 1.30 mol eq) in tetrahydrofuran (120.0 mL, 3.00 rel vol) was prepared at 20-25⁰C. Charged this solution into the above reaction mass containing (1), maintaining 20-25⁰C internal temperature, in about 10-15 min [Mild exotherm (3-4°C) observed]. Charged tetrahydrofuran (40.0 mL, 1.00 rel vol) as line wash. Stirred the resulting mass for 30 min. Meanwhile charged sodium triacetoxyborohydride (86.81 g, 84.81 g @100%, 0.4002 mol,

1.50 mol eq) into a 2.0 L four-necked jacketed vessel equipped with a mechanical stirrer, thermo well, condenser, nitrogen inlet, Huber unit for heating and cooling, 500.0 mL pressure equalising funnel and a distillation set-up. Charged tetrahydrofuran (280.0 mL, 7.00 rel vol) and stirred the mass for 5-10 min at 20-25⁰C. Cooled the resultant mass to 15- 17°C. Charged the above mixture of (1) and (2) into the cooled reaction mass, maintaining the internal temperature at 17±2°C. Tetrahydrofuran (40.0 mL, 1.00 rel vol) was charged as line wash. Stirred the reaction mass at 17±2°C for 1.0-1.5 h. Monitored the reaction by HPLC. Charged water (600.0 mL, 15.00 rel vol) to the reaction mass at 17±2°C [Mild exotherm (4°C) observed]. Raised the temperature of reaction mass to 35°C and stirred the resulting mass at 30-35⁰C for 5-10 min. Distilled out tetrahydrofuran from the reaction mass under vacuum (300-140 mbar) at 30-35⁰C (white solid precipitates out). Stirred the precipitated product at 20-25⁰C for 1.0 h. Filtered the product and washed the wet cake with water (600.0 mL, 15.00 rel vol). Deliquored the solid for 10-15 min. Unloaded the wet cake and dried it in a vacuum oven at 45±5°C till constant weight was obtained. Dry weight of the product was 75.84g @100%.

Yield: 88.78% w/w (based on HPLC assay), HPLC Purity: (3) - 99.2 %w/w, HPLC assay: 100.8 % w/w.

¹H NMR δ(_CD3₀D) 7.67 (2H, bs), 7.32 (2H, d), 3.55 (2H, s), 3.46-3.40 (4H, m), 2.45-2.39 (4H, m), 1.46 (9H, s).

MS (APCI+) 321.2 [M+H]⁺

The following method was used for LC/MS analysis:

Instrument Agilent 1100; Ionization source: APCI; Column - Zorbax Cis-column, 150 mm x 4.6 mm ID, 5μm particle size; Flow rate 1.0 ml/min; Wavelength 230 nm; Solvent A: 0.03 % TFA in water; Solvent B: 0.03% TFA in acetonitrile; Gradient 0 % - 45% B 8 min, 70% B 10 min, 95 % B 12 min, 95% B 15 min. Stage II: Preparation of tert-butyl 4-((3'-(6-fluoro-3-((ls,4s)-4-(6-fluoroimidazo[l,2- alpyridine-l-carboxamidoJcyclohexylJ-l^-dioxo-S^-dihydropyridoβ^-dlpyrimidin- l(2h)-yl)biphenyl-4-yl)methyl)piperazine-l-carboxylate (5)

Charged 6-fluoro-N-((ls,4s)-4-(6-fluoro-l-(3-iodophenyl)-2,4-dioxo-l,2- dihydropyrido[2,3-d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carbo xamide (4) (81.97 g, 80.00 g @100%, 0.125 mol, 1.00 mol eq) followed by 4-(4-(tert- butoxycarbonyl)piperazin-l-yl)methyl)phenylboronic acid (3) (42.42 g, 41.87 g @100%, 0.131 mol, 1.05 mol eq) into a 2.0 L six-necked jacketed vessel, equipped with a mechanical stirrer, condenser, temperature sensor and nitrogen inlet and Huber unit for heating and cooling at temperature 20-25⁰C. Charged methanol (1040.0 mL, 13.00 rel vol) and stir the mass for 3-5 min. Then charged acetonitrile (240.0 mL, 3.00 rel vol) and stirred the reaction mass for 3-5 min. Hunig's base (48.77 g, 48.29 g @100%, 65.73 mL, 0.374 mol, 3.0 mol eq) was then added followed by a line wash with methanol (80.0 mL, 1.00 rel vol). Stirred the resulting mass for 3-5 min. Charged Dichloro bis(tri-o-tolyl phosphine) palladium(II) [Pd-115] (0.489 g, 0.00062 mol, 0.005 mol eq) followed by methanol (80.0 mL, 1.00 rel vol) as line wash. Agitated the reaction mass for 3-5 min. Raised the temperature of the reaction mass to reflux (60-65⁰C) and heated the mass for 2 to 3 hrs till the reaction goes to completion as indicated by HPLC (Reaction mass turns clear solution). Filtered the reaction mass through celite bed to remove precipitated Palladium particle and washed the residue with methanol (80.0 mL, 1.0 rel vol). Charged the clear filtrate into another vessel (2.0 L six-necked jacketed reactor, equipped with a mechanical stirrer, condenser, temperature sensor, nitrogen inlet, distillation set up and Huber unit for heating and cooling. Methanol (80.0 mL, 1.00 rel vol) was charged as a line wash. Distilled off methanol- acetonitrile mixture azeotropically till the precipitation of solid was almost complete (distill to approx. 5 rel. vol). Charged methanol (400.0 mL, 5.0 rel vol) once again and distilled out once again till the precipitation is complete (distill to approx. 5 rel. vol). [Typical acetonitrile content after second distillation is 1.00-2.00% w/w]. Cooled the precipitated reaction mass to room temperature under stirring. Continued stirring at room temperature for another 2.0-2.5 hrs. Filtered the precipitated solid and washed the solid with methanol (160.0 mL, 2.0 rel vol). Transferred the wet solid into another vessel and slurried the solid in water (800.0 mL, 10.0 rel vol) at 50-55 ⁰C for 40-45 min. Cooled the contents of the reaction mass to 35-40 ⁰C. Filtered the precipitated solid and washed the wet cake with water (400.0 mL, 5.00 rel vol) and deliquored. Dried the wet cake in a vacuum oven at 40-45⁰C till constant weight of tert-butyl 4-((3'-(6-fluoro-3-((ls,4s)-4-(6- fluoroimidazo[l,2-a]pyridine-2-carboxamido)cyclohexyl)-2,4-dioxo-3,4- dihydropyrido[2,3-d]pyrimidin- 1 (2h)-yl) biphenyl-4-yl)methyl)piperazine- 1 -carboxylate (5) was obtained. Dried weight of the product was 92.13 g @100%.

Yield: 93.54% w/w (based on HPLC assay), HPLC Purity: (5) - 99.15 %, HPLC assay: 99.6 % w/w.

¹H NMR δ(_DM_S0-d6) 8.83-8.72 (IH, m), 8.58 (IH, d), 8.36 (IH, d), 8.31 (IH, dd), 7.82-7.55 (6H, m), 7.50-7.32 (4H, m), 4.98-4.82 (IH, m), 4.18 (IH, bs), 3.52 (2H, s), 3.41-3.23 (4H, m), 2.74-2.48 (2H, m), 2.41-2.22 (4H, m), 2.10-1.91 (2H, m), 1.82-1.59 (4H, m), 1.39 (9H, S).

MS (APCI+) 791.2 [M+H]⁺

The following method was used for LC/MS analysis:

Instrument Agilent 1100; Ionization source: APCI; Column - Waters symmetry shield RP18-column, 150 mm x 4.6 mm ID, 5μm particle size; Flow rate 1.0 ml/min; Wavelength 220 nm; Solvent A: 0.03% TFA in water; Solvent B: 0.03% TFA in acetonitrile; Gradient 0 % - 45% B 4 min, 70 % B 6 min, 95 %B 8 min, 95% B 15 min. Stage III: Preparation of 6-fluoro-n-((ls,4s)-4-(6-fluoro-2,4-dioxo-l-(4'-(piperazin-l- ylmethyl)-biphenyl-3-yl)-l,2-dihydropyrido[2,3-d]pyrimidin-3(4h)- yl)cyclohexyl)imidazo[ 1 ,2-a]pyridine-2-carboxamide (6)

(5)

(6)

MW 790 87

MW 690 76

Charged (5) (80.56 g, 80.00 g @100%, 0.101 mol, 1.00 mol eq) into a 2.0 L six-necked jacketed vessel, equipped with a mechanical stirrer, condenser, temperature sensor nitrogen inlet and Huber unit for heating and cooling. Charged purified water (400.0 mL, 5.00 rel vol). Stirred the resulting slurry for 10 minutes at 20-25⁰C. Raised the temperature of the reaction mass to 55-60⁰C and charged aqueous hydrochloric acid [Cone. HCl (35%w/w), 44.0 mL, 52.10 g, 18.44 g @100%, 0.505 mol, 5.0 mol eq, diluted with purified water (160.0 mL, 2.0 rel vol)] slowly at 55-60⁰C]. Water (80.0 mL, 1.0 rel vol) was charged as line wash. Agitated the contents of the reaction mass for 1-2 hrs. Monitored the reaction by HPLC. After the completion of reaction, cooled the contents of the reaction mass to 45- 50⁰C and extracted the reaction mass twice with toluene (240.0 mL, 3.0 rel vol) (some minor impurities will be selectively removed in toluene layer upon extraction). Separated the aqueous layer and filtered it over a celite bed to remove precipitated particles . Washed the celite bed with purified water (80.0 mL, 1.0 rel vol) and deliquored for about 5.0 min. Transferred the filtrate into another vessel and raised the temperature of the mass to 45- 50⁰C. Adjusted the pH of the filtrate by drop wise addition of sodium hydroxide solution (5.0% w/v, 420.0 mL, 5.25 rel vol) to pH = 10.0-10.5 at 45-50⁰C. Stirred the precipitated reaction mass for 30 min. Filtered the precipitated product at 45-50 ⁰C, washed the wet cake with purified water (640.0 mL, 8.0 rel vol) and deliquored for lhr. Dried the wet cake in a vacuum oven at 50±5 ⁰C, till constant weight of 6-fluoro-n-((ls,4s)-4-(6-fluoro-2,4- dioxo- 1 -(4'-(piperazin- 1 -ylmethyl)-biphenyl-3-yl)- 1 ,2-dihydropyrido[2,3-d] pyrimidin-

3(4h)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide (6) was obtained. Dried weight of the product was 59.16 g @100%.

Yield: 84.7% w/w (based on NMR assay), HPLC Purity: (6)- 99.25% (By HPLC), NMR Assay: 93.9% w/w (Moisture content - 3.9% w/w).

¹H NMR δ_(DMso₎ 8.83-8.72 (IH, m), 8.58 (IH, d), 8.36 (IH, s), 8.31 (IH, dd), 7.82-7.54 (6H, m), 7.49-7.31 (4H, m), 4.99-4.75 (IH, m), 4.18 (IH, bs), 3.46 (2H, s), 2.75-2.45 (7H, m), 2.40-2.20 (4H, m), 2.07-1.92 (2H, m), 1.82-1.58 (4H, m).

MS (APCI+) 691.2 [M+H]⁺

The following method was used for LC/MS analysis:

Instrument Agilent 1100; Ionization source: APCI; Column - Waters symmetry shield

RP18-column, 150 mm x 4.6 mm ID, 5μm particle size; Flow rate 1.0 ml/min; Wavelength 220 nm; Solvent A: 0.03 % TFA in water; Solvent B: 0.03% TFA in acetonitrile; Gradient 0% - 45% B 4 min, 70% B 6 min, 95 % B 8 min, 95% B 15 min.

A useful intermediate that can be used in the preparation of 6-fluoro-n-((ls,4s)-4-(6-fluoro- 2,4-dioxo-l -(4'-(piperazin- 1 -ylmethyl)-biphenyl-3-yl)- 1 ,2-dihydropyrido[2,3-d]pyrimidin- 3(4h)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide (6) is presented in Route A below.

Stage IV: Preparation of 6-fluoro-n-((ls,4s)-4-(6-fluoro-2,4-dioxo-l-(4'-(piperazin-l- ylmethyl)-biphenyl-3-yl)-l,2-dihydropyrido[2,3-d]pyrimidin-3(4h)- yl)cyclohexyl)imidazo [ 1 ,2-a] pyridine-2-carboxamide, (1 s)-(+)-l 0-camphor sulfonic acid salt (7)

Charged 6-fluoro-N-(( 1 s,4s)-4-(6-fluoro-2,4-dioxo- 1 -(4'-(piperazin- 1 -ylmethyl)-biphenyl- 3-yl)-l,2-dihydropyrido[2,3-d]pyrimidin-3(4h)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2- carboxamide (6) (42.23 g, 40.00 g @100%, 0.058 mol, 1.00 mol eq) into a 1.0 L four- necked round bottom flask equipped with a mechanical stirrer, thermo well, nitrogen inlet etc at 25-30⁰C. Charged ethyl acetate (600.0 ml, 15.00 rel vol). Agitated the contents of the reaction mass for 30 min (slight hazy solution obtained). Filtered the contents of the reaction mass over a celite bed (in order to remove any undissolved material). Washed the celite bed with ethyl acetate (80.0 ml, 2.0 rel vol). Transferred the clear filtrate to a 2.0 L six-necked jacketed vessel, equipped with a mechanical stirrer, 500.0 ml pressure equalising funnel, condenser, temperature sensor, nitrogen inlet and Huber unit for heating and cooling. Ethyl acetate (40.0 ml, 1.0 rel vol) was used as a line wash. Raised the temperature of the reaction mass to 45-50⁰C. Meanwhile (lS)-(+)-10-camphorsulphonic acid (13.04 g, 12.78 g @100%, 0.055 moles, 0.95 mol eq) was dissolved in ethyl acetate (400.0 ml, 10.00 rel vol) in a 1.0 L four necked round bottom flask equipped with a mechanical stirrer, thermo well and nitrogen inlet under agitation at 25-30⁰C. Filtered the resulting clear solution. Charged the solution of (lS)-(+)-10-camphorsulphonic acid into the reaction mass containing solution of (6) in about 1.0-1.5 h at 45-50⁰C (solid precipitates out). Ethyl acetate (40.0 ml, 1 rel vol) was used as a line wash. Stirred the precipitated reaction mass for 1.0-1.5 h. Filtered the product (7) at 45-50⁰C and washed the wet cake with ethyl acetate (120.0 ml, 3.0 rel vol). Deliquored the cake for 30 min. unloaded the solid and dried the material in a vacuum oven at 50-55⁰C, till constant weight of 6-fluoro-n-((l s,4s)-4-(6-fluoro-2,4-dioxo- 1 -(4'-(piperazin- 1 -ylmethyl)-biphenyl-3-yl)- l,2-dihydropyrido[2,3-d]pyrimidin-3(4h)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2- carboxamide,(lS)-(+)-10-camphorsulfonic acid salt (7) was obtained. Dried weight of the product was 51.56 g @100%.

Yield: 96.46% w/w (based on NMR assay), HPLC Purity: (7)- 99.0%, NMR assay: 97.29% w/w, Stoichiometry: Freebase: Camphor Sulphonic acid = 1.0: 0.98.

¹U NMR δccDsoD+DMSo-de) 8.65-8.57 (IH, m), 8.45 (IH, d), 8.35 (IH, s), 8.30 (IH, dd), 7.83-7.75 (IH, m), 7.74-7.58 (5H, m), 7.52-7.43 (2H, m), 7.42-7.32 (2H, m), 5.18-4.91 (IH, m), 4.31 (IH, s), 3.68 (2H, s), 3.30-3.18 (5H, m), 2.92-2.61 (8H, m), 2.42-2.28 (IH, m), 2.22-1.97 (4H, m), 1.96-1.70 (5H, m), 1.67-1.52 (IH, m), 1.49-1.35 (IH, m), 1.15 (3H, s), 0.87 (3H, s).

MS (APCI+) 691.2 [M+H]⁺

The following method was used for LC/MS analysis: Instrument Agilent 1100; Ionization source: APCI; Column - Waters symmetry shield

RP18-column, 150 mm x 4.6 mm ID, 5μm particle size; Flow rate 1.0 ml/min; Wavelength 220 nm; Solvent A: 0.03 % TFA in water; Solvent B: 0.03% TFA in acetonitrile; Gradient 5 % - 40% B 14 min, 70% B 18 min, 95% B 22 min, 95% B 30 min.

Preparation 6

Preparation of 6-Fluoro-7V-{c/s-4-[6-fluoro-2,4-dioxo-l-[4'-(piperazin-l- ylmethyl)biphenyl-3-yl] - 1 ,4-dihydropyrido [2,3-d] pyrimidin-3(2H)- yl]cyclohexyl}imidazo[l,2-«]pyridine-2-carboxamide as a tri-hydrochloric acid salt

Concentrated hydrochloric acid (2.2 mL) were added to a solution of 6-fluoro-Λ/-{cώ-4-[6- fluoro-2,4-dioxo-l-[4'-(piperazin-l-ylmethyl)biphenyl-3-yl]-l,4-dihydropyrido[2,3- J]pyrimidin-3(2H)-yl]cyclohexyl}imidazo[l,2-α]pyridine-2-carboxamide (1.6 g, 2.32 mmol) was dissolved in ethanol (20 mL). The resultant suspension was diluted with further ethanol (750 mL) and recrystallised in this solvent before being allowed to slurry 48 hours. The solid precipitate was then collected by filtration, washed with ethanol (200 mL), and dried in vacuo to give the title compound as a colourless solid (1.45 g).

¹H NMR (400 MHz, DMSO-d6) δ 9.89 - 9.59 (m, 2H), 8.84 (dd, IH), 8.60 (d, IH), 8.42 (s, IH), 8.34 (dd, IH), 7.86 - 7.59 (m, 9H), 7.54 - 7.38 (m, 2H), 4.88 (t, IH), 4.51 - 4.27 (m, 2H), 4.16 (s, IH), 3.97 - 3.34 (m, 8H), 2.69 - 2.55 (m, 2H), 2.02 (d, 2H), 1.79 - 1.60 (m, 4H).

Chloride Ion analysis: Found: Cl, 12.83% C₃₈H₃₆F₂N₈O₃-SHCl requires Cl, 13.20%

Preparation 7

Preparation of 6-fluoro-N-((ls,4s)-4-(6-fluoro-l-(3-iodophenyl)-2,4-dioxo-l,2- dihydropyrido [2,3-d] pyrimidin-3(4H)-yl)cyclohexyl)imidazo [1 ,2-a]pyridine-2-carbo xamide (4)

See Route B (below)

Preparation of 5-Fluoro-2-(3-iodophenylamino)nicotinic acid 2-Chloro-5-fluoro nicotinic acid (12.322 g; 70.19 mmol), p-toluenesulfonic acid monohydrate (6.817 g; 35.84 mmol), toluene (118.7 g) and 3-iodoaniline (13.993 g; 63.89 mmol) were charged to a 200 ml glass autoclave. The autoclave was immersed in a silicon oil bath that was at 14O⁰C. A clear solution was obtained when the internal temperature reached 81⁰C. At an internal temperature of 102⁰C, the contents of the autoclave were boiling and at this point the autoclave was sealed. The internal temperature rose to 127⁰C as the pressure increased (0.7 bar). After 40 minutes the internal temperature had increased to 134.4°C and the pressure had increased to 1.6 bar. The reaction was held at temperature and pressure for a further 28 hours. The reaction was cooled to 98°C and then the autoclave was vented. With an internal temperature at 92.5°C, water (199g) was charged over ca. 40 minutes and cooling was initiated. With the temperature at 38°C, the precipitated product was isolated by filtration. The cake was washed with toluene (32.7g) and water (95.9g) and the sucked dry to yield 39.53 g of wet product. The product was dried at 70⁰C at 50mbar to give 18.18g (79%) of dry product. Assay by NMR: 94.5%

Preparation of t-Butyl cis-4-(5-fluoro-2-(3-iodophenylamino)nicotinamido)cyclohexyl- carbamate

5-Fluoro-2-(3-iodophenylamino)nicotinic acid (94.5%; 15.00 g; 39.58 mmol), t-butyl cis-4-aminocyclohexylcarbamate (97.6%; 11.75 g; 53.51 mmol) and DMF (102.8 g) were charged to a round-bottomed flask with overhead stirring. Triethylamine (27.70 g; 273.74 mmol) was added resulting in a clear brown solution with a temperature of 18.5⁰C. The solution was cooled to -1.1⁰C and T3P (2,4,6-tripropyl-[l,3,5,2,4,6]trioxatriphosphinane 2,4,6-trioxide) in THF (50%; 45.04 g; 70.78 mmol) was dosed over 3 hours. The reaction mixture was heated to 21.8⁰C and then water (44.26 g) was charged allowing the mixture to exotherm to 33.2⁰C. the temperature was adjusted to 38.8°C and water (7.33 g) was charged, maintaining a constant temperature (39.2-39.9°C) for 1 hour. Gradual cooling to 33.2°C over 1 hour gave the start of crystallisation. After 35 minutes a thick slurry had formed. Additional water (43.58 g) was charged over 45 minutes and then gentle cooling was applied over 1 hour to bring the temperature to 17.8°C. The product was collected by filtration, and the filter cake was rinsed with aqueous DMF (40%; 41.5 g) and water (117.0 g) to give 37.88 g of wet product. The product was dried in a vacuum oven at 36 mbar / 67⁰C to give 20.47 g (93%) of dry product. Assay by NMR: 98.7% Water content (KF): 0.49%

Preparation of *-Butyl cis-4-(6-fluoro-l-(3-iodophenyl)-2,4-dioxo-l,2- dihydropyrido [2,3-d] pyrimidin-3(4H)-yl) cyclohexylcarbamate t-Butyl cis-4-(5 -fluoro-2-(3 -iodophenylamino)nicotinamido)cyclohexylcarbamate

(98.7%; 16.01 g; 28.5 mmol), l,l '-carbonyldiimidazole (9.27 g; 57.2 mmol) and NMP (147.0 g) were charged to a round-bottomed flask with overhead stirring. A clear solution was obtained. The flask was cooled to 0.9⁰C and sodium hydride in mineral oil (60%; 814 mg) was charged in 7 portions over a total time period of 3 hours 45 minutes whilst maintaining the temperature at less than 2.5°C. After the addition of each portion of sodium hydride the gas evolution was allowed to diminish before charging the next portion. The reaction was then stirred for 50 minutes before water (17.62 g) was carefully added whilst keeping the temperature at or below 10⁰C. The reaction was heated to 44.1°C and then water (47.58 g) was added at a constant temperature of 45⁰C; this took ca. 50 minutes. At this point a granular precipitation was observed. The mixture was cooled to 17.7°C over 1 hour 45 minutes. The product was collected by filtration and the filter cake was rinsed with aqueous NMP (50%; 36.5 g) and water (162.5 g) to give 21.0 g of wet product. Assay by NMR: 78.6% Assay compensated yield: 99.8 %

Preparation of 6-Fluoroimidazo[l,2-a]pyridine-2-carboxylic acid hydrochloride

2-Amino-5-fluoropyridine (93%; 14.00 g; 116.14 mmol) and toluene (101.4 g) was charged to a round-bottomed flask with overhead stirring and heated to 50.6⁰C; 2-amino-5- fluoropyridine was almost dissolved. Toluene (22.8 g) was charged. With the mixture at 6O⁰C, a clear solution with some dense solids was obtained. The solution was vacuum filtered and the filter was rinsed with toluene (19.6 g). The collected solids (impurities in the raw material) amounted to 1.02g. The filtrate (153.65 g) was further diluted with toluene to a total weight of 167.3 g. Ethyl bromopyruvate (84%; 34 g; 146.45 mmol) was charged over 35 minutes at an internal temperature of 43-48⁰C; the addition was slightly exothermic. A thick slurry containing black oily particles was formed. The reaction was checked for completion by NMR and then cooling was applied. Aqueous hydrochloric acid (35-37%; 60.49 g) was charged over 11 minutes whilst keeping the temperature at 2.5- 6.O⁰C; the addition was slightly exothermic. A clear amber two-phase system was formed with the lower layer being much darker than the upper layer. The mixture was heated to 18⁰C and the upper layer was separated and discarded. The lower layer was heated towards boiling (85.1⁰C). The mixture was heated to 107.2⁰C over 1 hour collecting the distillate. After heating for a further 80 minutes, whilst continuing to collect the distillate, NMR analysis showed that the ester hydrolysis was complete. The temperature was decreased and at 63°C crystallisation started. The mixture was re-heated to 67.4°C and then slowly cooled. At 61⁰C gross precipitation occurred and the material could not be re- dissolved on heating to 71°C. Acetone (39.13 g) was charged over 9 minutes; temperature decreased from 69⁰C to 47⁰C. The mixture was cooled further over 40 minutes from 47⁰C to 28⁰C. Methyl ethyl ketone (161.4 g) was charged over 5 minutes; temperature decreased from 28⁰C to 23⁰C. the mixture was cooled to -1.5⁰C over 10 minutes and then held for an hour. With the temperature now at -3.6⁰C the product was isolated by filtration. The filter cake was rinsed with methyl ethyl ketone (61.6 g) and sucked dry to give 32.23 g of wet product. The product was dried in a vacuum oven overnight at 73⁰C and 4 mbar to give 21.93g (87%) of dry product.

Assay by NMR: 96.2% (calculated as the mono-hydrochloride) Assay compensated yield: 83.9 % Water content (Karl Fischer): 0.33%

Preparation of 6-Fluoro-N-(cis-4-(6-fluoro- 1 -(3-iodophenyl)-2,4-dioxo- 1 ,2- dihydropyrido[2,3-d]pyrimidin-3(4H)-yl) cyclohexyl)imidazo[ 1 ,2-a]pyridine-2- carboxamide hydrochloride t-Butyl cis-4-(6-fluoro-l-(3-iodophenyl)-2,4-dioxo-l,2-dihydropyrido[2,3- d]pyrimidin-3(4H)-yl) cyclohexylcarbamate (78.6% assay; 16.01 g; 21.7 mmol) and acetonitrile (76.0 g) were charged to a round-bottomed flask with overhead stirring; internal temperature dropped to 16.3⁰C. Aqueous hydrochloric acid (35-37%; 11.8Ig; 116.6 mmol) was added whereupon the temperature rose to 21.7⁰C. The suspension was heated to 34⁰C. Additional aqueous hydrochloric acid (35-37%; 1.42g; 14.1 mmol) was charged to form a cloudy suspension. After 20 minutes the cloudy suspension turned clear with oily drops within it. The reaction mixture was distilled under vacuum (maximum temperature 35°C) in several cycles. After each cycle acetonitrile was added to replace the solvent that had been distilled; in total 230.9 g of acetontrile were added over four cycles. In total, 21O g of distillate was collected. The residue was a yellow to light brown oil containing a small amount of crystals (water content by Karl Fischer = 1.21%). With the temperature at 23.4°C, triethylamine (18.4O g; 181.8 mmol) was added over 10 minutes to give a mobile light-coloured slurry. The mixture was cooled to 0.5⁰C and then held overnight under nitrogen atmosphere (for convenience). With the internal temperature at 12.6⁰C, 6-fluoroimidazo[l,2-a]pyridine-2-carboxylic acid hydrochloride (96% assay; 4.91 g; 21.8 mmol) was charged to the vessel causing the temperature to rise from 15.6⁰C to 20.8⁰C; small lumps were formed which dissipated on stirring and a smooth slurry was obtained. In process control by HPLC at 220 nm showed 49.2 area-% of 6-fluoroimidazo[l,2-a]pyridine-2-carboxylic acid hydrochloride and 50.4 area-% of 3-(cis-4-aminocyclohexyl)-6-fluoro-l-(3-iodophenyl) pyrido[2,3-d]pyrimidine- 2,4(1 H,3H)-dione. The reaction was cooled to 10.3⁰C. T3P in THF (50%; 27.3 g; 42.9 mmol) was charged over 1 hour keeping the temperature below 12°C. The temperature was then increased to 5O⁰C. Water (25.0 g) was added over 40 minutes whilst maintaining the temperature at 50⁰C. After a further 1 hour 45 minutes the mixture was cooled to 27.6°C over 90 minutes. The mixture was left to cool further and was stirred overnight (16 hours). With the temperature at 18⁰C, water (101.1 g) was slowly dosed in over 1 hour (initially at 0.5 ml/minute). With the temperature at 16°C the product was isolated by vacuum filtration. The filter cake was rinsed with aqueous acetonitrile (40%; 27.0 g) and water (84.Ig). The filter cake was sucked dry to give 15.40 g of wet product. The wet product was homogenized in a closed container and 3.653 g was dried in a vacuum cabinet overnight at 71⁰C to give 2.684 g of dry product.

Calculated yield = 15.4Og x ^2MAS = π _{31 g} (₈₁ o_/o)

3.653g

Assay by NMR: 97.0% Assay compensated yield: 78.8 % Water content (Karl Fisher): 1.1 %

The present invention will now be further explained by reference to the following illustrative Examples.

Example 1

Evaluation of compound activity on intra-alveolar neutrophil migration after aerosol challenge with lipopolysaccharride (LPS) in the CRLrCD rat LPS challenge in CRL:CD rats causes an influx of inflammatory cells into the lungs. Rats are challenged either with an aerosol of 0.9% w/v saline or O.lmg/mL LPS in 0.9% saline for 30 min or an intratracheal dose of 0.1-10μg/kg. This is repeated up to 8 times according to the experimental protocol. Rats are dosed with vehicle, standard compound or test compound by the appropriate route and frequency at various time points before and after challenge depending upon the experimental protocol. Test compound groups could either be the same compound at different doses or single doses of different compounds or a combination of the two. Test compounds are given by intraperitoneal, intravenous or subcutaneous injection or by inhalation or intratracheal administration.

The rats are euthanized at various time points after challenge depending upon the nature of the study, but typically 4hr after LPS challenge with ImL pentobarbitone sodium. A tracheotomy is performed and a cannula inserted. The airway is then lavaged using 3 mL sterile PBS at room temperature. The PBS is left in the airway for 10 seconds before being removed. The PBS containing cells is placed into a 15 mL centrifuge tube on ice. This process is repeated three times.

An aliquot of BAL fluid is removed and counted on Sysmex (Sysmex UK, Milton Keynes). Cytospin slides are prepared by adding a 100 μl aliquot of BAL fluid into cytospin funnels in a Shandon Cytospin3 operated at 700 rpm for 5 min. Slides are stained on the Hema-Tek-2000 automatic slide stainer, using Wright-Giemsa stain and typically, 200 cells are counted under a microscope. Cells are classified as eosinophils, neutrophils and mononuclear cells (mononuclear cells included monocytes, macrophages and lymphocytes) and are expressed as a percentage of the total count.

Example 2

Evaluation of lung function in anaesthetised guinea-pigs

Male Dunkin-Hartley guinea-pigs (300-60Og) are weighed and dosed with either vehicle or compound in an appropriate vehicle according to the experimental protocol via the intratracheal route under recoverable gaseous anaesthesia (5% halothane in oxygen).

Following dosing, the animals are administered supplemental oxygen and monitored until full recovery. Typically a dose volume of 0.5 mL/kg is used for the intratracheal route. In a dose response study, animals are dosed with compound or vehicle two hours prior to the administration of histamine. Test compound groups could either be the same compound at different doses or single doses of different compounds or a combination of the two.

The guinea-pigs are anaesthetised with pentobarbitone (1 mL/kg of 60 mg/mL solution intraperitoneally) approximately 30 minutes prior to the first bronchoconstrictor administration. The trachea is cannulated (Portex intravenous cannula, 200/300/070 (orange) or 200/300/060 (yellow)) and the animal ventilated using a constant volume respiratory pump (Harvard Rodent Ventilator model 683) at a rate of 60 breath/min and a tidal volume of 5 ml/kg. A jugular vein is cannulated (Portex intravenous catheter 200/300/010 (green)) for the administration of histamine or maintenance anaesthetic (0.1 mL of pentobarbitone solution, 60 mg/mL, as required).

The animals are then transferred to a Flexivent System (SCIREQ, Montreal, Canada) in order to measure airway resistance. The animals are ventilated (quasi-sinusoidal ventilation pattern) at 60 breaths/min at a tidal volume of 5 mL/kg. A positive end expiratory pressure of 2-3 CmH₂O is applied. Respiratory resistance is measured using the Flexivent "snapshot" facility (1 second duration, 1 Hz frequency). Once stable baseline resistance value has been obtained the animals are given histamine dihydrochloride or methacholine in ascending doses (histamine; 0.5, 1, 2, 3 and 5μg/kg, i.v., methacholine; 3, 10 and 30 μg/kg, i.v.) at approximately 4-minute intervals via the jugular catheter. After each administration of histamine the peak resistance value is recoreded. Guinea pigs are euthanised with approximately 1.OmL pentobarbitone sodium (Euthatal) intravenously after the completion of the lung function measurements. Percentage bronchoprotection produced by a compound is calculated at each dose of histamine as follows:

% changeR_veh - % changeR_cmpd

% bronchoprotection = ^■

% changeR_v 'eh

Where % change R_veh is the mean of the maximum percentage change in airway resistance in the vehicle treated group.

Example 3

Evaluation of Compounds on Antigen induced Eosinophilia in Ovalbumin Sensitised Brown Norway Rats On day 0 of the study Brown Norway rats are given a subcutaneous injection of 500 μg ovalbumin adsorbed onto 100 mg aluminium hydroxide in 0.4 mL saline in two distinct sites, approximately 0.2 mL per site. Day 14 and 15 following sensitisation the rats are challenged with aerosolised ovalbumin for 15 minutes. The rats are placed in groups of 10 in an acrylic box (internal dimensions 320mm wide x 320mm deep x 195 mm high, 2OL volume). 8mL of 10 mg/mL ovalbumin in 0.9% saline, or 0.9% saline alone, is placed in each of two jet nebulizers (Sidestream®, Profile Respiratory Systems Ltd.). Compressed air at 6 L/min is passed through each nebulizer and the output of the nebulizers is passed into the box containing the rats.

Rats are dosed via the appropriate route with vehicle, standard compound or test compound at various time points before and after challenge depending upon the experimental protocol. Rats are euthanised with 0.5 mL pentobarbitone sodium (Euthatal) intraperitoneally at various times after challenge. A tracheotomy is performed and the trachea cannulated. The airway is then lavaged using 3 mL sterile PBS at room temperature. The PBS is left in the airway for 10 seconds before being removed. The PBS containing cells is placed into a 15 mL centrifuge tube on ice. This process is repeated three times. The final volume recovered is recorded. An aliquot of BAL fluid is removed and counted using a Sysmex (Sysmex UK, Milton Keynes).

Cytospin slides are prepared by adding a 100 μl aliquot of BAL fluid into cytospin funnels in a Shandon Cytospin 3 operated at 700 rpm for 5 min. Slides are stained on the Hema- Tek-2000 automatic slide stainer, using Wright-Giemsa stain and typically, 200 cells are counted under a microscope. Cells are classified as eosinophils, neutrophils and mononuclear cells. Mononuclear cells included monocytes, macrophages and lymphocytes.

Example 4

Evaluation on the effect of compound on lung function and BAL-neutrophilia following acute smoke exposure in the mouse BALB/c or C57BL6/J mice undergo whole body exposure to main stream smoke (50 min/ 12 cigarettes) and fresh air once or twice a day for 1-9 days. Mice are dosed via the appropriate route with vehicle, standard compound or test compound at various time points before and after challenge depending upon the experimental protocol. On the final day of the experiment, mice are either killed with euthatal 0.2 ml i.p. and broncho-aveolar lavage fluid obtained for analysis of white blood cell infiltration (as described above) or lung function is assessed using a Flexivent System (SCIREQ, Montreal, Canada). Alternatively lung mechanics are measured using a forced manoeuvres system (EMMS).

Mice are anaesthetised with pentobarbitone (1/lOdilution at a dose volume of 1 mL/kg intraperitoneally). The trachea is cannulated and the animal transferred to the Flexivent System where they are ventilated (quasi-sinusoidal ventilation pattern) at a rate of 150 breath/min and a tidal volume of 10 ml/kg in order to measure airways resistance. Respiratory resistance is measured using the Flexivent "snapshot" facility (1 second duration, 1 Hz frequency). Mice are euthanised with approximately 0.5mL pentobarbitone sodium (Euthatal) intravenously after the completion of the lung function measurements.

Example 5

Evaluation of bronchodilator activity in the guinea-pig isolated tracheal ring preparation.

Guinea-pigs (300-60Og) are killed by cervical dislocation and the trachea removed. After clearing the adherent connective tissue, the trachea is cut into four ring segments (2-3 cartilage rings in width) and suspended in 10ml organ baths containing modified Krebs' solution (gassed with 5% CO₂, 95% O₂ at 37°C). The tracheal rings are attached to an isometric force transducer for the measurement of isometric tension. The tissues are washed and a force of Ig was applied to each tissue. The rings are precontracted with methacholine (1 μM) and a cumulative (10^"9M - 10^"5M) isoprenaline concentration effect curve is constructed. Responses are expressed as a percentage relaxation of the methacholine induced contraction. The rings are washed and a second concentration of methacholine (lμM) is added. Once the contraction has reached a plateau isoprenaline or the compound under investigation is added until a maximum effective dose is reached. Data are collected using the AD Instruments chart4forwindows software, which measures the maximum tension generated at each concentration of agonist and the response expressed as percentage relaxation.

Example 6

Inhibition of lipopolysaccharride (LPS)-induced TNFα production in human peripheral blood mononuclear cells. Compound A is: N-[2-(Diethylamino)ethyl]-N-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-l,3- benzothiazol-7-yl)ethyl] amino } ethyl)-3 -[2-(I -naphthyl)ethoxy]propanamide dihydrobromide

Compound B is: 6-fluoro-N-((ls,4s)-4-(6-fluoro-2,4-dioxo-l-(4'-(piperazin-l-ylmethyl)- biphenyl-3-yl)- 1 ,2-dihydropyrido[2,3-d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[ 1 ,2- a]pyridine-2-carboxamide trihydrochloride

Compound-induced inhibition of LPS stimulated TNFα production was determined by pre- incubating human isolated peripheral blood mononuclear cells (PBMCs) with the PDE4 inhibitor Compound B (lOpM - 100 nM) alone and in the presence of the β₂ adrenergic agonist Compound A (lOOpM-lμM) for 30 mins at 37°C. The cells were then incubated with LPS (lμg/mL) for 24 hours at 37°C to induce TNFα production. At the end of the incubation period, the plates were centrifuged (300g, 10 minutes) and 50 μL of the culture supernatant was analysed to quantify the TNFα released using a Flourescence-linked immunosorbance assay (FLISA) assay kit (R&D Systems). Fluorescence levels were read on an FMAT plate reader. Inhibition curves were fitted using a 4-parameter logistic equation in a non-linear curve fitting routine and activity was expressed as pICso. Compound B (10OnM) produced a 73 % inhibition of LPS-induced TNFα production. In the presence of 1OnM, 10OnM and lμM Compound A, the maximum inhibition of TNFα production was 75, 87 and 95%, respectively (n=2). The mean data for the combination of the compounds from the two human donors are shown below in Figure 3 and Table 1. Compound A alone inhibited TNFα production by 46% at 1 μM. The mean data for Compound A are shown below in Figure 4.

Figure 3: Effect of the combination of β2 adrenergic agonist (Compound A) and PDE4 antagonist (Compound B) on LPS stimulated TNFα production from human PBMC

+ Vehicle

+ 10OpM Compound A + 1 nM Compound A + 1OnM Compound A + 10OnM Compound A +1 μM Compound A

log [Compound B](M)

Figure 4: Effect of the β2 adrenergic agonist Compound A on LPS stimulated TNFα production from human PBMC (mean data from 2 human donors).

log [Compound A](M)

Isobole analysis of the data at 40% inhibition of TNFα production showed that the interaction between the PDE4 inhibitor Compound B and the β₂ adrenergic agonist Compound A was synergistic. This is shown below in Figure 5.

Figure 5. Isobole analysis of the effect of a combination of the PDE4 inhibitor Compound B and the β2 adrenergic agonist Compound A on LPS stimulated TNFα production from human PBMC (mean data from 2 human donors).

Isobole analysis at 40% inhibition

[Compound A] (nM) ROUTE A

(reduce weight by 30%) then MEK and cool further

An optionally protected compound of formula (I) can be prepared from this intermediate using metal mediated coupling, for example with a suitable substituted boronic acic

T₃P = Propane phosphonic acid anhydride MEK = methyl ethyl ketone ptsa = p-toluenesulphonic acid ROUTE B

AZD5793 DihydrolPBr

C₅H₇BrO₃ 2-amιne C₁₀H₁₂BrFN₂O₃

195 01 C₅H₅FN₂ 307 11

112 10

Claims

1. A pharmaceutical product comprising, in combination, a first active ingredient which 6-fluoro-N-(( 1 s,4s)-4-(6-fluoro-2,4-dioxo- 1 -(4'-(piperazin- 1 -ylmethyl)-biphenyl-3- yl)-l,2-dihydropyrido[2,3-d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-

2-carboxamide or a salt thereof, and a second active ingredient selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; an antioxidant; a β₂ adrenoceptor agonist; a CCRl antagonist; a chemokine antagonist (not CCRl); a corticosteroid; a CRTh2 antagonist; a DPI antagonist; an Histone Deacetylase Inducer; an IKK2 inhibitor; a COX inhibitor; a lipoxygenase inhibitor; a leukotriene receptor antagonist; a MABA compound; an MPO inhibitor; a muscarinic antagonist; a p38 inhibitor; a PPARγ agonist; a protease inhibitor; a Statin; a thromboxane antagonist; a vasodilator; or, an ENAC blocker (Epithelial Sodium-channel blocker).

2. A pharmaceutical product as claimed in claim 1 wherein the first active ingredient is in the form of a salt which is a hydrochloride, hydrobromide (such as dihydrobromide), trifluoroacetate, sulphate, phosphate, acetate, fumarate, maleate, tartrate, lactate, citrate, pyruvate, succinate, oxalate, methanesulphonate, p- toluenesulphonate, bisulphate, benzenesulphonate, ethanesulphonate, malonate, xinafoate, ascorbate, oleate, nicotinate, saccharinate, adipate, formate, glycolate, L- lactate, D-lactate, aspartate, malate, L-tartrate, D-tartrate, stearate, 2-furoate, 3- furoate, napadisylate (naphthalene- 1, 5 -disulfonate or naphthalene- 1 -(sulfonic acid)- 5 -sulfonate), edisylate (ethane- 1 ,2-disulfonate or ethane- 1 -(sulfonic acid)-2- sulfonate), isethionate (2-hydroxyethylsulfonate), 2-mesitylenesulphonate, 2- naphthalenesulphonate, 2,5-dichlorobenzenesulphonate, D-mandelate, L-mandelate, cinnamate, benzoate, adipate, esylate, malonate, mesitylate (2- mesitylenesulphonate), napsylate (2-naphthalenesulfonate), camsylate (camphor- 10- sulphonate), formate, glutamate, glutarate, glycolate, hippurate (2-

(benzoylamino)acetate), orotate, xylate (p-xylene-2-sulphonate), pamoic (2,2'- dihydroxy-l,r-dinaphthylmethane-3,3'-dicarboxylate), palmitate or furoate.

3. A pharmaceutical product as claimed in claim 1 wherein the first active ingredient is in the form of a salt which is a camsylate salt (camphor- 10-sulphonate salt, for example (lS)-(+)-10-Camphorsulfonic acid salt).

4. A pharmaceutical product as claimed in claim 1, 2 or 3 wherein the second active ingredient selected from: a β₂ adrenoceptor agonist; a MABA compound; or a muscarinic antagonist.

5. Use of a product according to any one of claims 1 -4 in therapy.

6. Use of a product according to any one of claims 1 to 4 in the manufacture of a medicament for the treatment of a respiratory disease.

7. Use according to claim 5, wherein the respiratory disease is chronic obstructive pulmonary disease.

8. 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 as defined in claim 1 ;

(b) a (therapeutically effective) dose of a second active ingredient which is as defined in claim 1 ; to a patient in need thereof.

9. A kit comprising a preparation of a first active ingredient which is 6-fluoro-N- (( 1 s,4s)-4-(6-fluoro-2,4-dioxo- 1 -(4'-(piperazin- 1 -ylmethyl)-biphenyl-3 -yl)- 1 ,2- dihydropyrido[2,3-d]pyrimidin-3(4H)-yl)cyclohexyl)imidazo[l,2-a]pyridine-2- carboxamide or a salt thereof, a preparation of a second active ingredient which is selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; an antioxidant; β₂ adrenoceptor agonist; a CCRl antagonist; a chemokine antagonist (not CCRl); a corticosteroid; a CRTh2 antagonist; a DPI antagonist; an Histone Deacetylase Inducer; an IKK2 inhibitor; a COX inhibitor; a lipoxygenase inhibitor; a leukotriene receptor antagonist; a MABA compound; an MPO inhibitor; a muscarinic antagonist; a p38 inhibitor; a PPARγ agonist; a protease inhibitor; a Statin; a thromboxane antagonist; a vasodilator; or, an ENAC blocker (Epithelial Sodium-channel blocker), and optionally instructions for the simultaneous, sequential or separate administration of the preparations to a patient in need thereof.

10. A pharmaceutical composition comprising, in admixture, a first active ingredient which is 6-fluoro-N-(( 1 s,4s)-4-(6-fluoro-2,4-dioxo- 1 -(4'-(piperazin- 1 -ylmethyl)- biphenyl-3-yl)- 1 ,2-dihydropyrido[2,3-d]pyrimidin-3(4H)- yl)cyclohexyl)imidazo[l,2-a]pyridine-2-carboxamide or a salt thereof, and a second active ingredient which is selected from: a non-steroidal Glucocorticoid Receptor

(GR Receptor) Agonist; an antioxidant; β₂ adrenoceptor agonist; a CCRl antagonist; a chemokine antagonist (not CCRl); a corticosteroid; a CRTh2 antagonist; a DPI antagonist; an Histone Deacetylase Inducer; an IKK2 inhibitor; a COX inhibitor; a lipoxygenase inhibitor; a leukotriene receptor antagonist; a MABA compound; an MPO inhibitor; a muscarinic antagonist; a p38 inhibitor; a PPARγ agonist; a protease inhibitor; a Statin; a thromboxane antagonist; a vasodilator; or, an ENAC blocker (Epithelial Sodium-channel blocker).

11. A process for the preparation of a compound of formula (I):

with a compound of formula (III):

R¹

HN (Ml)

2

-R under reductive conditions, at ambient temperature.

12. An intermediate compound of formula (IV):

13. An intermediate compound that is : 5-Fluoro-2-(3-iodophenylamino)nicotinic acid; tert-Butyi cis-4-(5-fluoro-2-(3-iodophenylamino)nicotinamido)cyclohexyl- carbamate; tert-Butyl cis-4-(6-fluoro- 1 -(3-iodophenyl)-2,4-dioxo- 1 ,2-dihydropyrido[2,3- d]pyrimidin-3 (4H)-yl) cyclohexylcarbamate; 3-(cis-4-Aminocyclohexyl)-6-fluoro- 1 -(3-iodophenyl) pyrido[2,3-d]pyrimidine-

2,4(1 H,3H)-dione;

2-Ethoxycarbonyl-6-fluoro-2-hydroxy-2,3-dihydro-lH-imidazo[l,2-a]pyridin-4- yliumbromide; or,

6-Fluoro-N-(cis-4-(6-fluoro-l-(3-iodophenyl)-2,4-dioxo-l,2-dihydropyrido[2,3- d]pyrimidin-3(4H)-yl) cyclohexyl)imidazo[ 1 ,2-a]pyridine-2-carboxamide hydrochloride.