[go: up one dir, main page]

WO2008104776A1 - Combinations of beta-2-adrenoceptor agonistic benzothiazolone - Google Patents

Combinations of beta-2-adrenoceptor agonistic benzothiazolone Download PDF

Info

Publication number
WO2008104776A1
WO2008104776A1 PCT/GB2008/000667 GB2008000667W WO2008104776A1 WO 2008104776 A1 WO2008104776 A1 WO 2008104776A1 GB 2008000667 W GB2008000667 W GB 2008000667W WO 2008104776 A1 WO2008104776 A1 WO 2008104776A1
Authority
WO
WIPO (PCT)
Prior art keywords
amino
chloro
oxy
hydroxy
piperidin
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/GB2008/000667
Other languages
French (fr)
Inventor
Elaine Bridget Cadogan
Stephen Connolly
David John Nicholls
Alan Young
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
AstraZeneca UK Ltd
AstraZeneca AB
Original Assignee
AstraZeneca UK Ltd
AstraZeneca AB
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by AstraZeneca UK Ltd, AstraZeneca AB filed Critical AstraZeneca UK Ltd
Publication of WO2008104776A1 publication Critical patent/WO2008104776A1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/425—Thiazoles
    • A61K31/428—Thiazoles condensed with carbocyclic rings

Definitions

  • 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).
  • respiratory diseases for example chronic obstructive pulmonary disease (COPD) or asthma.
  • COPD chronic obstructive pulmonary disease
  • 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.
  • ARDS Acute Respiratory Distress Syndrome
  • COPD Chronic Obstructive Pulmonary Disease
  • 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 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.
  • Corticosteroids also known as glucocorticosteroids or glucocorticoids
  • glucocorticosteroids are potent antiinflammatory 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.
  • steroids may lead to therapeutic effects
  • 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 ⁇ 2 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.
  • Example 25 in PCT/SE2006/000981 produces what is referred to herein as Polymorphic Form A of the dihydrobromide salt of 7-[(l/?)-2-( ⁇ 2-[(3- ⁇ [2-(2-Chlorophenyl)ethyl]amino ⁇ - propyl)thio]ethyl ⁇ amino)- 1 -hydroxyethyl]-4-hydroxy- 1 ,3-benzothiazol-2(3H)-one.
  • the compound and its salts show at least a 10-fold selectivity of ⁇ 2 adrenoceptor agonism over adrenergic ⁇ lD, adrenergic ⁇ l and dopamine D2 activities.
  • Combination products comprising a ⁇ 2 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.
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-( ⁇ 2-[(3- ⁇ [2-(2-)
  • a second active ingredient selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; an antioxidant; a CCRl antagonist; a chemokine antagonist (not CCRl); a corticosteroid; a CRTh2 antagonist; a DPI antagonist; an ⁇ istone Deacetylase Inducer; an IKK2 inhibitor; a COX inhibitor; a lipoxygenase inhibitor; a leukotriene receptor antagonist; an MPO inhibitor; a muscarinic antagonist which is Aclidinium bromide, Glycopyrrolate (such as R 5 R-, R,S-,
  • S 5 R-, or S,S-glycopyrronium bromide Oxitropium bromide, Pirenzepine, telenzepine or Tiotropium bromide; a p38 inhibitor; a PDE inhibitor; a PPAR ⁇ agonist; a protease inhibitor; a Statin; a thromboxane antagonist; a vasodilator; or, an ENAC blocker (Epithelial Sodium-channel blocker).
  • the first and second active ingredients are in forms suitable for oral administration.
  • a suitable salt of 7-[(17?)-2-( ⁇ 2-[(3- ⁇ [2-(2-Chlorophenyl)ethyl]amino ⁇ propyl)thio]- ethyl ⁇ amino)-l-hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3H)-one is, for example, a hydrochloride, 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, nic
  • 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 or 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 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- pyrrolidiny 1 ⁇ amino]-2-hy droxypropoxy ⁇ -4-fluoropheny l)acetamide, N-(2 ⁇ (2S)-3 [ ⁇ (3 S)- 1 - [(4-chlorophenyl)methyl]-3-pyrrolidinyl ⁇ amino]-2-hydroxypropoxy ⁇ -4- fluoropheny l)acetamide, N-(2- ⁇ (2S)-3 - [ 1 - ⁇ (4-chlorobenzoy l)-4-piperidinyl ⁇ amino]-2- hydroxypropoxy ⁇ -4-hydroxyphenyl)acetamide, (2- ⁇ [(2S)-3- ⁇ [(2R,5S)- 1
  • a chemokine antagonist (other than a CCRl antagonist), for example, 656933 (N-(2- bromophenyl)-N'-(4-cyano-l ⁇ -l,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-(triflu
  • 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.
  • 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, OC 1768, OC2125, OC2184, OC499, OCD9101, I 0 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- i 5 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. 0
  • 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 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, Rl 503, SC-80036, SCIO469, 6-chloro-5-[[(25,5/?)-4-[(4-fluorophenyl)methyl]- 2,5-domethyl- 1 -piperazinyl]carbonyl]-iV,N, 1 -trimethyl- ⁇ -oxo- 1 H-indole-3-acetamide,0 VX702 or VX745 (5-(2,6-dichlorophenyl)-2-(phenylthio)
  • a PDE Inhibitor such as a PDE4 inhibitor, for example, 256066, Arofylline (3-(4- chlorophenyl)-3,7-dihydro-l -propyl- lH-Purine-2,6-dione), AWD 12-281 (N-(3,5-dichloro- 4-pyridinyl)-l-[(4-fluorophenyl)methyl]-5-hydroxy- ⁇ -oxo-lH-indole-3-acetamide), BAY19-8004 (Bayer), CDC-801 (Calgene), Celgene compound (( ⁇ R)- ⁇ -(3,4- dimethoxypheny I)- 1 ,3 -dihydro- 1 -oxo-2H-isoindole-2-propanamide), Cilomilast (cis-4- cyano-4-[3-(cyclopentyloxy)-4-methoxyphenyl]-cyclohexanecarboxylic
  • PDE4 inhibitors include 256066, Arofylline (3-(4-chlorophenyl)-3,7- dihydro-1 -propyl- lH-Purine-2,6-dione), AWD 12-281 (N-(3,5-dichloro-4-pyridinyl)-l-[(4- fluorophenyl)methyl]-5-hydroxy- ⁇ -oxo-lH-indole-3-acetamide), BAYl 9-8004 (Bayer), CDC-801 (Calgene), Celgene compound (( ⁇ R)- ⁇ -(3,4-dimethoxyphenyl)-l,3-dihydro-l- oxo-2H-isoindole-2-propanamide), Cilomilast (cis-4-cyano-4-[3-(cyclopentyloxy)-4- methoxyphenyl]-cyclohexanecarboxylic acid), a compound in WO2006098353 from Ky
  • a PPAR ⁇ 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 Doxycycline).
  • 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, Tre
  • An ENAC Episomal Sodium-channel blocker
  • Amiloride Benzamil, Triamterene, 552-02, PSA14984, PSA25569, PSA23682 or AER002. All the above active ingredients may be in the form of a solvate, e.g. a hydrate.
  • the present invention provides a pharmaceutical product comprising the first and second active ingredients in admixture.
  • the pharmaceutical product may, for example, be a kit comprising a preparation of the first active ingredient and a preparation of the second active ingredient and, optionally, instructions for the simultaneous, sequential or separate administration of the preparations to a patient in need thereof.
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-( ⁇ 2-[(3- ⁇ [2-(2-)
  • a second active ingredient 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 which is Aclidinium bromide, Glycopyrrolate (such as R 5 R-, R,S-, S,R-, or S,S-glycopyrronium bromide), Oxitropium bromide, Pirenzepine, telenzepine or
  • Tiotropium bromide a p38 inhibitor; or, a PDE inhibitor.
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ afnino)- 1 -hydroxyethyl]-4-hydroxy- 1 ,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient which is a nonsteroidal Glucocorticoid Receptor (GR) Agonist for example, a compound disclosed in WO 2006/046916.
  • GR Glucocorticoid Receptor
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient which is a CCRl antagonist, 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)-l-[[2- [(aminocarbonyl)amino]-4-chlorophenoxy]acetyl]-4-[(4-fluoropheny
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient which is a chemokine antagonist (not CCRl), for example, 656933 (N-(2-bromophenyl)-N'-(4-cyano- 1 ⁇ - 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, Cyclon
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l ⁇ )-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a corticosteroid, 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 Mo
  • the corticosteroid is selected from budesonide, fluticasone propionate, fluticasone fruoate mometasone furoate, beclomethasone dipropionate or butixocort propionate ester.
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(li?)-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a corticosteroid, for example, Budesonide, Fluticasone Furoate or Fluticasone propionate.
  • a corticosteroid for example, Budesonide, Fluticasone Furoate or Fluticasone propionate.
  • the corticosteroid is budesonide.
  • Budesonide and its preparation is described, for example, in Arzneistoff-Forschung (1979), 29 (11), 1687-1690, DE 2,323,215 and US 3,929,768.
  • Presently available formulations of budesonide are marketed under the tradename 'Entocort ®'.
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(li?)-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is an IKK2 inhibitor, for example, 2- ⁇ [2-(2-Methylamino-pyrimidin-4-yl)-l ⁇ -indole-5-carbonyl]- amino ⁇ -3-(phenyl-pyridin-2-yl-amino)-propionic acid.
  • a first active ingredient which is 7-[(li?)-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ amino)-l-
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(li?)-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ amino)- 1 -hydroxyethyl]-4-hydroxy- 1 ,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a muscarinic antagonist, for example, Aclidinium bromide, Glyc ⁇ pyrrolate (such as R 5 R-, R,S-, S 5 R-, or S,S-glycopyrronium bromide), Oxitropium bromide, Pirenzepine, telenzepine or Tiotropium bromide.
  • a first active ingredient which is 7-[(li?)-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇
  • the muscarinic receptor antagonist is a long acting muscarinic receptor antagonist, i.e. a muscarinic receptor antagonist with activity that persists for more than 12 hours.
  • long acting muscarinic receptor antagonists include tiotropium bromide.
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(li?)-2-( ⁇ 2-[(3- ⁇ [2-(2-)
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ amino)- 1 -hydroxyethyl]-4-hydroxy- 1 ,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a p38 inhibitor, 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(3 ⁇ )- pyrimidinone), Array-797, AZD6703, Doramapimod, KC-706, PH 797804, Rl 503, SC- 80036, SCIO469
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(li?)-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a PDE Inhibitor: such as a PDE4 inhibitor ⁇ for example, 256066, Arofylline (3-(4-chlorophenyl)- 3,7-dihydro-l-propyl-l ⁇ -Purine-2,6-dione), AWD 12-281 (N-(3,5-dichloro-4-pyridinyl)- 1 -[(4-fluorophenyl)methyl]-5-hydroxy- ⁇ -oxo- 1 H-indole-3-acetamide
  • WO2006098353 from Kyowa Hakko Kogyo Co. Ltd. Japan, 2-(3,5-dichloro-4-pyridinyl)- 1 -(7-methoxyspiro[ 1 ,3-benzodioxole-2, 1 '-cyclopentan]-4-yl)ethanone (CAS number 185406-34-2)), Compound from Pfizer (2-(3,4-difluorophenoxy)-5-fluoro-N-[cis-4-[(2- hydroxy-5-methylbenzoyl)amino]cyclohexyl]-)-3-pyridinecarboxamide), Compound from Pfizer (2-(3,4-difluorophenoxy)-5-fluoro-N-[cis-4-[[2-hydroxy-5- (hydroxymethyl)benzoyl]amino]cyclohexyl]-3-pyridinecarboxamide,), CT2820, GPD- 1116, Ibudilast, IC 485, KF 3133
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a PDE4 inhibitor, for example, 256066, Arofylline (3-(4-chlorophenyl)-3,7-dihydro-l -propyl- 1 ⁇ - Purine-2,6-dione), AWD 12-281 (N-(3,5-dichloro-4-pyridinyl)-l-[(4- fluorophenyl)methyl]-5-hydroxy- ⁇ -oxo- 1 ⁇ -indole-3-acetamide), BAY 19-8004 (Bayer),
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a PDE4 5 inhibitor, for example AWD 12-281 (N-(3,5-dichloro-4-pyridinyl)-l-[(4- fluoropheny l)methyl]-5-hydroxy- ⁇ -oxo- 1 ⁇ -indole-3-acetamide) or roflumilast.
  • a first active ingredient which is 7-[(l/?)-2-( ⁇ 2-[(3- ⁇ [2-(2- Chlorophenyl)ethyl]amino ⁇ propyl)thio]e
  • the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l ⁇ )-2-( ⁇ 2-[(3- ⁇ [2-(2- i o Chlorophenyl)ethy l]amino ⁇ propyl)thio]ethyl ⁇ amino)- 1 -hydroxyethyl]-4-hydroxy- 1,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is roflumilast.
  • the present invention provides a kit comprising a preparation of a first active ingredient which is 7-[(li?)-2-( ⁇ 2-[(3- ⁇ [2-(2- i 5 Chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one or a salt thereof, a preparation of a second active ingredient selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; an antioxidant; 20 a CCRl antagonist; a chemokine antagonist (not CCRl); a corticosteroid; a CRTh2 antagonist; a DPI antagonist; 5 an ⁇ istone Deacetylase Inducer; an IKK2 inhibitor; a COX inhibitor; a lipoxygenase inhibitor; a leukotriene receptor antagonist; 30 an MPO inhibitor; a GR Re
  • Tiotropium bromide Tiotropium bromide; a p38 inhibitor; a PDE inhibitor; a PPAR ⁇ 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.
  • ENAC blocker Epitophelial Sodium-channel blocker
  • 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.
  • simultaneous is meant that the active ingredients are in admixture or they could be in separate chambers of the same inhaler.
  • 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 e.g. 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 colourant.
  • pharmaceutically acceptable ingredients 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 colourant.
  • the first and second active ingredients are administered via a single pharmaceutical composition (that is, they are in admixture). Therefore, the present invention further provides a pharmaceutical composition comprising, in admixture, a first active ingredient which is 7-[(li?)-2-( ⁇ 2-[(3- ⁇ [2-(2-)
  • the pharmaceutical composition further comprises a pharmaceutically acceptable adjuvant, diluent or carrier.
  • 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.
  • 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.
  • the first active ingredient is administered via inhalation.
  • the dose of the first active ingredient will generally be in the range of from 0.1 microgram ( ⁇ g) to 5000 ⁇ g, 0.1 to 1000 ⁇ g, 0.1 to 500 ⁇ g, 0.1 to 100 ⁇ g, 0.1 to 50 ⁇ g, 0.1 to 5 ⁇ g, 5 to 5000 ⁇ g, 5 to 1000 ⁇ g, 5 to 500 ⁇ g, 5 to 100 ⁇ g, 5 to 50 ⁇ g, 5 to 10 ⁇ g, 10 to 5000 ⁇ g, 10 to 1000 ⁇ g, 10 to 500 ⁇ g, 10 to 100 ⁇ g, 10 to 50 ⁇ g, 20 to 5000 ⁇ g, 20 to 1000 ⁇ g, 20 to 500 ⁇ g, 20 to 100 ⁇ g, 20 to 50 ⁇ g, 50 to 5000 ⁇ g, 50 to 1000 ⁇ g, 50 to 500 ⁇ g, 50 to 100 ⁇ g, 100 to 5000 ⁇ g, 100 to 1000 ⁇ g or 100
  • the second active ingredient is administered by inhalation.
  • the dose of the second active ingredient will generally be in the range of from 0.1 microgram ( ⁇ g) to 5000 ⁇ g, 0.1 to 1000 ⁇ g, 0.1 to 500 ⁇ g, 0.1 to 100 ⁇ g, 0.1 to 50 ⁇ g, 0.1 to 5 ⁇ g, 5 to 5000 ⁇ g, 5 to 1000 ⁇ g, 5 to 500 ⁇ g, 5 to 100 ⁇ g, 5 to 50 ⁇ g, 5 to 10 ⁇ g, 10 to 5000 ⁇ g, 10 to 1000 ⁇ g, 10 to 500 ⁇ g, 10 to 100 ⁇ g, 10 to 50 ⁇ g, 20 to 5000 ⁇ g, 20 to 1000 ⁇ g, 20 to 500 ⁇ g, 20 to 100 ⁇ g, 20 to 50 ⁇ g, 50 to 5000 ⁇ g, 50 to 1000 ⁇ g, 50 to 500 ⁇ g, 50 to 100 ⁇ g, 100 to 5000 ⁇ g, 100 to 1000 ⁇ g or 100 to
  • 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.
  • 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.
  • 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.
  • a metered dose inhaler device may be used to administer the first and second 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 is a hydrocarbon, chlorofluorocarbon or hydrofluoroalkane (e.g. heptafluoroalkane) propellant, or mixture of any such propellant, for example in a pressurised metered dose inhaler (pMDI).
  • a preferred propellant is P 134a or P227, each of which may be used alone or in combination with other propellants and/or surfactant and/or other excipients.
  • 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.
  • 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).
  • Dry powder inhalers may be used to administer the active ingredients, alone or in combination with a pharmaceutically acceptable carrier, in the later case either as a finely divided powder or as an ordered mixture.
  • the dry powder inhaler may be single dose or multi-dose and may utilise a dry powder or a powder-containing capsule.
  • Metered dose inhaler, nebuliser and dry powder inhaler devices are well known and a variety of such devices are available.
  • the pharmaceutical product 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 NSAID-induced) and dust-induced asthma, both intermittent and persistent and of all severities, and other causes of airway hyper-responsiveness; chronic obstructive pulmonary disease (COPD); bronchitis, including infectious and eosinophilic bronchitis and chronic 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 s infections;
  • 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 is particular chronic obstructive pulmonary disease or asthma.
  • the present invention still further provides a method of treating a respiratory disease which comprises simultaneously, sequentially or separately administering: (a) a therapeutically effective dose of a first active ingredient as defined above; and,0 (b) a therapeutically effective dose of a second active ingredient as defined above; to a patient in need thereof.
  • the term “therapy” also includes “prophylaxis” unless there are specific indications to the contrary.
  • the terms “therapeutic” and 5 "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 by0 genetic testing or screening to be particularly susceptible to developing the condition or disorder.
  • Figure 1 Shows the onset times for roflumilast (l ⁇ M), Compound A (0.InM) and Compound A (0.InM) in the presence of roflumilast (l ⁇ M) in guinea pig trachea in vitro.
  • Aluminium hydride was prepared by the drop- wise addition of a solution of sulphuric acid (8.40 mL) in dry THF (60 mL) to a stirred solution of 1.0M lithium aluminium hydride in THF (314 mL), at 0-10 0 C, under a nitrogen atmosphere. After stirring at 5 0 C for 30 minutes, a solution of l-chloro-2-[(£)-2-nitrovinyl]benzene (12.83 g) in dry THF (160 mL) was added dropwise maintaining the internal temperature between O 0 C and 1O 0 C. When the addition was complete the reaction was heated at reflux for 5 minutes.
  • tert-Butyl allyl[2-(2-chlorophenyl)ethyl]carbamate (31.0 g) was mixed with 2-mercaptoethanol (7.37 mL), and AIBN (1.15 g), and stirred at 65 0 C for 45 minutes. The mixture was cooled and more mercaptoethanol (1 mL) and AIBN (200 mg) added. The mixture was then heated at 65 0 C for a further 30 minutes. The material was purified by 0 silica column chromatography, loading the material in 20% ethyl acetate in isohexane, then eluting with 20% ethyl acetate in isohexane, changing to 50%, to give the desired material (31.94 g).
  • the reaction mixture was diluted with ethyl acetate, washed with water, then IN HCl, then saturated sodium bicarbonate solution, dried over anhydrous magnesium sulfate, filtered and the solvents removed in vacuo.
  • the material was purified by silica column chromatography eluting with 20% ethyl acetate in isohexane to give the desired material (12.43 g).
  • Type A material (Example 2) was placed into a vial, to which was added water (ImI). The mixture was left to stir at room temperature in a capped vial for one week. The resulting suspension was then centrifuged and the solid collected and left to dry overnight in a fume hood.
  • Fumaric acid 120.39 mg was added to a suspension of 7-[(li?)-2-( ⁇ 2-[(3- ⁇ [2-(2- chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ -amino)- 1 -hydroxyethyl]-4-hydroxy- 1 ,3- benzothiazol-2(3H)-one (0.5 g) in methanol (5 mL). The mixture was then stirred at room temperature for 2 h. The solvent was removed in vacuo and the residue was suspended in ethyl acetate (20 mL) and stirred at room temperature for 48 h. The title compound was isolated by filtration, washed with ethyl acetate (5 mL) and dried in vacuo to leave a noncrystalline product (0.59 g).
  • Citric acid (199.27 mg) was added to a suspension of 7-[(l/?)-2-( ⁇ 2-[(3- ⁇ [2-(2- chlorophenyl)ethyl]amino ⁇ propyl)thio]ethyl ⁇ -amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one (0.5 g) in methanol (5 mL). The mixture was sonicated then stirred at room temperature for 2 h. The solvent was removed in vacuo and the residue was suspended in diethyl ether (20 mL) and stirred at room temperature for 1 h. The title compound was isolated by filtration, washed with diethyl ether (5 mL) and dried in vacuo to leave a non-crystalline product.
  • Phosphoric acid (119.58 mg) was added to a suspension of 7-[(U?)-2-( ⁇ 2-[(3- ⁇ [2-(2- chlorophenyl)ethyl]amino ⁇ propyl)thio]ethy 1 ⁇ -amino)- 1 -hydroxy ethy l]-4-hydroxy- 1,3- benzothiazol-2(3H)-one (0.5 g) in methanol (5 mL). The mixture was then stirred at room temperature for 1 h. The solvent was removed in vacuo and the residue was suspended in diethyl ether (20 mL) and stirred at room temperature for 16 h. The solvent had evaporated so the residue was treated with more diethyl ether (5 mL). The title compound was isolated by filtration, washed with diethyl ether (5 mL) and dried in vacuo to leave a non-crystalline product (0.47g).
  • H292 cells were grown in 225cm2 flasks incubator at 37°C, 5% CO 2 in RPMI medium containing, 10% (v/v) FBS (foetal bovine serum) and 2 mM L-glutamine.
  • the culture media was removed and cells were washed twice with 100 ⁇ L assay buffer and replaced with 50 ⁇ L assay buffer (HBSS solution containing 1OmM HEPES pH7.4 and 5 mM glucose). Cells were rested at room temperature for 20 minutes after which time 25 ⁇ L of rolipram (1.2 mM made up in assay buffer containing 2.4% (v/v) dimethylsulphoxide) was added. Cells were incubated with rolipram for 10 minutes after which time Compound A was added and the cells were incubated for 60 minutes at room temperature. The final rolipram concentration in the assay was 300 ⁇ M and final vehicle concentration was 1.6% (v/v) dimethylsulphoxide. The reaction was stopped by removing supernatants, washing once with 100 ⁇ L assay buffer and replacing with 50 ⁇ L lysis buffer. The cell monolayer was frozen at -80°C for 30 minutes (or overnight). AlphaScreenTM cAMP detection
  • the concentration of cAMP (cyclic adenosine monophosphate) in the cell lysate was determined using AlphaScreenTM methodology. The frozen cell plate was thawed for 20 minutes on a plate shaker then 10 ⁇ L of the cell lysate was transferred to a 96- well white 5 plate. 40 ⁇ L of mixed AlphaScreenTM detection beads pre-incubated with biotinylated cAMP, was added to each well and the plate incubated at room temperature for 10 hours in the dark. The AlphaScreenTM signal was measured using an EnVision spectrophotometer (Perkin-Elmer Inc.) with the recommended manufacturer's settings. cAMP concentrations were determined by reference to a calibration curve determined in the same experiment
  • Membranes were prepared from human embryonic kidney 293 (HEK293) cells expressing recombinant human ⁇ lo receptor. These were diluted in Assay Buffer (5OmM HEPES, ImM EDTA, 0.1% gelatin, pH 7.4) to provide a final concentration of membranes that gave a clear window between maximum and minimum specific binding. 5
  • Assay Buffer 5OmM HEPES, ImM EDTA, 0.1% gelatin, pH 7.4
  • the plates were incubated for 2 hours at room temperature and then filtered onto PEI coated GF/B filter plates, pre-soaked for 1 hour in Assay Buffer, using a 96-well plate Tomtec cell harvester. Five washes with 250 ⁇ L wash buffer (5OmM HEPES, ImM EDTA, pH 7.4) were performed at 4°C to remove unbound radioactivity. The plates were dried then sealed from underneath using Packard plate sealers and MicroScint-0 (50 ⁇ L) was added to each well. The plates were sealed (TopSeal A) and filter-bound radioactivity was measured with a scintillation counter (TopCount, Packard BioScience) using a 3-minute counting protocol.
  • wash buffer 250 ⁇ L wash buffer
  • MicroScint-0 50 ⁇ L
  • Membranes containing recombinant human adrenergic beta 1 receptors were obtained from Euroscreen. These were diluted in Assay Buffer (5OmM HEPES, ImM EDTA, 12OmM NaCl, 0.1% gelatin, pH 7.4) to provide a final concentration of membranes that gave a clear window between maximum and minimum specific binding.
  • Assay Buffer 5OmM HEPES, ImM EDTA, 12OmM NaCl, 0.1% gelatin, pH 7.4
  • the plates were incubated for 2 hours at room temperature and then filtered onto PEI coated GF/B filter plates, pre-soaked for 1 hour in Assay Buffer, using a 96-well plate Tomtec cell harvester. Five washes with 250 ⁇ L wash buffer (5OmM HEPES, ImM EDTA, 12OmM NaCl, pH 7.4) were performed at 4°C to remove unbound radioactivity. The plates were dried then sealed from underneath using Packard plate sealers and MicroScint-0 (50 ⁇ L) was added to each well. The plates were sealed (TopSeal A) and filter-bound radioactivity was measured with a scintillation counter (TopCount, Packard BioScience) using a 3-minute counting protocol.
  • a scintillation counter TopCount, Packard BioScience
  • B 0 Total specific binding was determined by subtracting the mean NSB from the mean maximum binding. NSB values were also subtracted from values from all other wells. These data were expressed as percent of B 0 .
  • Compound concentration-effect curves (inhibition of [ 125 I]-Iodocyanopindolol binding) were determined using serial dilutions typically in the range 0.1 nM to 10 ⁇ M. Data was fitted to a four parameter logistic equation to determine the compound potency, which was expressed as pIC 5 o (negative log molar concentration inducing 50% inhibition of [ 125 I]-Iodocyanopindolol binding). A result is shown in Table 1 below.
  • Membranes containing recombinant human Dopamine Subtype D2s receptors were obtained from Perkin Elmer. These were diluted in Assay Buffer (5OmM HEPES, ImM EDTA, 12OmM NaCl, 0.1% gelatin, pH 7.4) to provide a final concentration of membranes that gave a clear window between maximum and minimum specific binding.
  • Assay Buffer 5OmM HEPES, ImM EDTA, 12OmM NaCl, 0.1% gelatin, pH 7.4
  • the plates were incubated for 2 hours at room temperature and then filtered onto PEI coated GF/B filter plates, pre-soaked for 1 hour in Assay Buffer, using a 96-well plate Tomtec cell harvester. Five washes with 250 ⁇ L wash buffer (5OmM HEPES, ImM EDTA, 12OmM NaCl, pH 7.4) were performed at 4°C to remove unbound radioactivity. The plates were dried then sealed from underneath using Packard plate sealers and MicroScint-0 (50 ⁇ L) was added to each well. The plates were sealed (TopSeal A) and filter-bound radioactivity was measured with a scintillation counter (TopCount, Packard BioScience) using a 3-minute counting protocol.
  • a scintillation counter TopCount, Packard BioScience
  • Example 1 The present invention will now be further explained by reference to the following illustrative Examples.
  • Example 1 The present invention will now be further explained by reference to the following illustrative Examples.
  • LPS lippopolvsaccharride
  • CRL:CD rats 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 0.1-1 O ⁇ g/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.
  • Cytospin slides are prepared by adding a 100 ⁇ l 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 compound activity on intra-alveolar neutrophil migration after aerosol challenge with lippopolysaccharride (LPS) in the guinea-pig.
  • Male Dunkin-Hartley guinea-pigs 300-60Og are placed into open fronted guinea-pig holding cones attached at random around a cylindrical aerosol chamber. Guinea-pigs are held in the challenge cones and exposed to an aerosol of vehicle, or LPS at concentrations of 0.1-30 ⁇ g/ml in 0.9%saline per group Aerosols are generated using 2 jet nebulisers per column with a flow rate of 12 L/m. 10ml of the challenge agent is placed into each nebuliser. Alternatively animals receive an intratracheal dose of 0.1-10 ⁇ g/kg. This is repeated up to 8 times according to the experimental protocol.
  • LPS lippopolysaccharride
  • Guinea-pigs 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.
  • Challenged guinea-pigs are killed by anaesthesia overdose (0.5ml Euthetal i.p.) at 4h-24h post challenge. The lungs are then lavaged.
  • HBSS Hanks Buffered Salt Solution
  • EDTA EDTA -free
  • the lavaging is performed with gentle massaging of the chest to ensure appropriate agitation of the fluid in the lungs.
  • the washes are harvested into a 15ml conical, polypropylene centrifuge tube, an aliquot of BAL fluid is removed and counted on Sysmex (Sysmex UK, Milton Keynes).
  • Cytospin slides are prepared by adding a 100 ⁇ l 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 3 Evaluation of compound activity on intra-alveolar neutrophil migration after aerosol challenge with lippopolysaccharride (LPS) in the mouse.
  • Male C57BL/6/J or BALB/C mice (20-35g) are placed in Perspex exposure boxes in groups of up to 20 and exposed to an aerosol of either 0.3 mg/ml LPS or 0.9% w/v saline.
  • the LPS (Sigma, E.Coli, Ref L-3755, Serotype 026:B6, Lot no. 11 lk4078) is made up in 0.9% w/v saline.
  • An aerosol is generated using two jet nebulisers operated at a flow rate of 12 L/min (6L/min for each nebuliser) for 15 min.
  • animals receive an intratracheal dose of 0.1-1 O ⁇ g/kg. This may be repeated up to 8 times.
  • mice 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.
  • mice are killed with an overdose of Euthatal i.p 30 minutes, l-24hr after LPS challenge.
  • the trachea is cannulated (Portex intravenous cannula) and the airways lavaged with 3 x 0.3ml of Isoton II (Beckman Coulter Ref. 8448011 Lot no.25775).
  • Isoton II Beckman Coulter Ref. 8448011 Lot no.25775
  • lOO ⁇ l of the BALF is added to a cytospin funnel and spun, using a ThermoShandon Cytospin model 3 or 4, at 700 rpm for 5 min.
  • Cells on the slide are stained on the Hema-Tek-2000 automatic slide stainer, using Wright-Giemsa stain and differential cell counts carried out to differentiate eosinophils, neutrophils and lymphomononuclear cells (including monocytes, macrophages and lymphocytes). Typically, 200 cells are counted per slide and each cell type expressed as a percentage of the total count. BALF total white cell count is measured using a Sysmex (Sysmex UK, Milton Keynes).
  • mice 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 CmH 2 O is applied.
  • Respiratory resistance is measured using the Flexivent "snapshot" facility (1 second duration, 1 Hz frequency).
  • the animals are given histamine dihydrochloride or methacholine in ascending doses (Histamine; 0.5, 1, 2, 3 and 5 ⁇ g/kg, i.v., methacholine; 3, 10 and 30 ⁇ g/kg, i.v.) at approximately 4-minute intervals via the jugular catheter. After each administration of histamine the peak resistance value is recorded. 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:
  • % change R veh is the mean of the maximum percentage change in airway resistance in the vehicle treated group.
  • 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 ⁇ l 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.
  • mice 20-25g male BALB/c mice are sensitized to ovalbumin by i.p administration of 100 ⁇ g of grade V ovalbumin (Sigma) adsorbed onto lmg of aluminium hydroxide gel mixture (Fisher Scientific UK) in 0.3 ml saline. Groups of mice are pre-dosed with compound if required, a minimum of two weeks after sensitization. They are then dosed daily for 1-8 days as study protocol specified, with test compound or 0.25 ml vehicle.
  • mice are placed in perspex chambers (20x11x1 lcm, 10 mice max./chamber) and administered an aerosol challenge of 20mg ml '1 ovalbumin for 36 min (8 ml for 18 min followed by another 8 ml for 18 min). Aerosol delivery is achieved using a DeVilbiss jet nebulizer with a flow rate of 61 min "1 . 24h after the last dose the mice are killed with euthatal 0.2 ml i.p.
  • trachea is cannulated using a pink luer mount Portex cannula cut to lcm and the lungs are lavaged using 3 washes of ImI of Isoton IL.
  • lOO ⁇ l of the BALF is added to a cytospin funnel and spun, using a ThermoShandon Cytospin model 3 or 4, at 700 rpm for 5 min.
  • Cells on the slide are stained on the Hema-Tek-2000 automatic slide stainer, using Wright-Giemsa stain and differential cell counts carried out to differentiate eosinophils, neutrophils and lymphomononuclear cells (including monocytes, macrophages and lymphocytes). Typically, 200 cells are counted per slide and each cell type expressed as a percentage of the total count. BALF total white cell count is measured using a Sysmex (Sysmex UK, Milton Keynes).
  • 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.
  • 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).
  • SCIREQ Flexivent System
  • EMMS forced manoeuvres system
  • 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 8 Evaluation of bronchodilator activity in the guinea pig isolated tracheal ring preparation: onset measurements.
  • Guinea pigs (300-50Og) were killed by cervical dislocation and the trachea was isolated.
  • the trachea was cut into segments 2-3 cartilage rings in width and suspended in 10ml organ baths in modified Krebs' solution (mM; NaCl, 90; NaHCO 3 , 45; KCl, 5; MgSO 4 JH 2 O, 0.5; Na 2 HPO 4 .2H 2 O, 1 ; CaCl 2 , 2.25; glucose, 10; pH 7.4 gassed with 5% CO 2 , 95% O 2 at 37°C).
  • the tracheal rings were attached to an isometric force transducer for the measurement of isometric tension.
  • the tissues were washed and a force of Ig was applied to each tissue.
  • the rings were contracted with methacholine (1 ⁇ M). Once the contraction had reached a plateau, vehicle (0.01% DMSO in distilled H 2 O), compound A (0.1 nM), roflumilast ( 1 ⁇ M) or a combination of compound A (0.1 nM) and roflumilast (l ⁇ M) was added and the tissue left until the response had reached a plateau. Data were collected using the Chart 4 software (ADInstruments, Charlgrove, UK). The time to 90% of the maximum effect of compound A (onset time) was measured for each tissue and expressed in min (mean ⁇ s.e.mean).

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention provides a pharmaceutical product, kit or composition comprising a first active ingredient which is 7-[(1R)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)-thio]ethyl}amino)-1-hydroxyethyl]-4-hydroxy-1,3-benzothiazol-2(3H)-one or a salt thereof, and a second active ingredient selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; an antioxidant; 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; an MPO inhibitor; a muscarinic antagonist which is Aclidinium bromide, Glycopyrrolate, Oxitropium bromide, Pirenzepine, telenzepine or Tiotropium bromide; a p38 inhibitor; a PDE 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

COMBINATIONS OF BETA-2 -ADRENOCEPTOR AGONISTIC BENZOTHIAZOLONE
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 antiinflammatory 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.
7-[(l/?)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l- hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3H)-one and its dihydrobromide, ditrifluoroacetate and di-acetate salts are β2 adrenoceptor agonists and are disclosed in PCT/SE2006/000981 (now published as WO2007/027134). Example 25 in PCT/SE2006/000981 produces what is referred to herein as Polymorphic Form A of the dihydrobromide salt of 7-[(l/?)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}- propyl)thio]ethyl} amino)- 1 -hydroxyethyl]-4-hydroxy- 1 ,3-benzothiazol-2(3H)-one. The compound and its salts show at least a 10-fold selectivity of β2 adrenoceptor agonism over adrenergic αlD, adrenergic βl and dopamine D2 activities.
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 the 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 7-[(l/?)-2-({2-[(3-{[2-(2-
Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one or a salt thereof, and a second active ingredient selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; an antioxidant; a CCRl antagonist; a chemokine antagonist (not CCRl); a corticosteroid; a CRTh2 antagonist; a DPI antagonist; an Ηistone Deacetylase Inducer; an IKK2 inhibitor; a COX inhibitor; a lipoxygenase inhibitor; a leukotriene receptor antagonist; an MPO inhibitor; a muscarinic antagonist which is Aclidinium bromide, Glycopyrrolate (such as R5R-, R,S-,
S5R-, or S,S-glycopyrronium bromide), Oxitropium bromide, Pirenzepine, telenzepine or Tiotropium bromide; a p38 inhibitor; a PDE inhibitor; a PPARγ agonist; a protease inhibitor; a Statin; a thromboxane antagonist; a vasodilator; or, an ENAC blocker (Epithelial Sodium-channel blocker).
In one particular aspect of the present invention, the first and second active ingredients are in forms suitable for oral administration. A suitable salt of 7-[(17?)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]- ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3H)-one is, for example, a hydrochloride, 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 and 2- naphthalenesulphonate.
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 or 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 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- pyrrolidiny 1 } amino]-2-hy droxypropoxy } -4-fluoropheny l)acetamide, N-(2 { (2S)-3 [ { (3 S)- 1 - [(4-chlorophenyl)methyl]-3-pyrrolidinyl}amino]-2-hydroxypropoxy}-4- fluoropheny l)acetamide, N-(2- {(2S)-3 - [ 1 - { (4-chlorobenzoy l)-4-piperidinyl } amino]-2- hydroxypropoxy }-4-hydroxyphenyl)acetamide, (2- { [(2S)-3- { [(2R,5S)- 1 -(4-chlorobenzyl)- 2,5-dimethylpiperidin-4-yl]amino}-2-hydroxy-2-methylpropyl]oxy}-4-fluorophenyl)acetic acid, (2-{[(2S)-3-{[(3S,4R)-l-(4-chlorobenzyl)-3-methylpiperidin-4-yl]amino}-2-hydroxy- 2-methylpropy 1] oxy } -4-fluoropheny l)acetic acid, (2- { [(2S)-3- { [(3 R,4R)- 1 -(4- chlorobenzyl)-3-methylpiperidin-4-yl]amino}-2-hydroxy-2-methylpropyl]oxy}-4- fluorophenyl)acetic acid, (2-{[(2S)-3-{[(2R,4S,5S)-l-(4-chlorobenzyl)-2,5- dimethylpiperidin-4-yl]amino}-2-hydroxy-2-methylpropyl]oxy}-4-fluorophenyl)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)-l-(4-chlorobenzyl)-2,5- dimethy lpiperidin-4-y 1] amino } -2-hydroxy-2-methy lpropyl]oxy } -4-fluoropheny l)acetic acid, Methyl (2-{[(2S)-3-{[l-(4-chlorobenzyl)piperidin-4-yl]amino}-2- hydroxypropy l]oxy } -4-fluoropheny l)propanoate, N-[2-({2S}-3-[(l- [4-chlorobenzy 1] -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-chlorobenzy 1] -4-piperidiny l)amino] -2-hy droxy-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 - [( 1 - [4-chlorobenzy l]-4-piperidinyl)amino]-2-hydroxy-2- methylpropoxy)-4-hydroxyphenyl] propaneamide, (2- { [(25)-3- { [ 1 -(4- chlorobenzyl)piperidin-4-yl]amino}-2-hydroxy-2-methylpropyl]oxy}-4- fluorophenyl)methanesulfonic acid, N-5-chloro-(2-{(2S)-3-[l-{(4-chlorobenzyl)-4- piperidinyl } amino] -2-hy droxypropoxy}-4-hydroxypheny I)-N' -cyclopropyl-urea, N-(2- { (2 S)-3- [ 1 - {(4-chlorobenzy l)-4-piperidiny 1 } amino]-2-hydroxypropoxy } -pheny I)-N' -ethyl- urea, (2S)-l-(2-ethylphenoxy)-3[(l-[4-chlorobenzyl]4-piperidinyl)amino]propan-2-ol, (2S)- 1 -[2-(-hydroxyethyl)phenoxy]-2-methyl-3 [( 1 -[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-lΗ,3H-spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-4- hydroxyphenyl)acetamide, N-(2-{[(2S)-3-(5-chloro-l'H-spiro[l,3-benzodioxole-2,4'- piperidin]- 1 '-yl)-2-hydroxypropyl]oxy } -4-hydroxyphenyl)acetamide, 2- { [(2S)-3-(5-chloro- l'H,3H-spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-4-hydroxy-N- methylbenzamide, 2- { [(2S)-3-(5-chloro- 1 Η,3 H-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-yl)- 2-hydroxypropy l]oxy } -4-hydroxybenzoic acid, N-(2- { [(2 S)-3 -(5-chloro- 1 Η,3 H-spiro [2- benzofuran- 1 ,4'-piperidin] - 1 '-y l)-2-hydroxypropy l]oxy } -4-hydroxypheny l)acetamide; 2-{ [(2S)-3-(5-chloro-l Η,3H-spiro[2-benzofuran- 1 ,4'-piperidin]- 1 '-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[l -benzofuran-2,3'-pyrrolidin]- l'-yl]-2-hydroxypropyl}oxy)-4- hydroxyphenyljacetamide, N-(2-{[(2S)-3-(5-chloro-l'H,3H-spiro[l-benzofuran-2,4'- piperidin]-l'-yl)-2-hydroxypropyl]oxy}-4-hydroxyphenyl)urea, 4-fluoro-2-{[(2S)-3-(5- fluoro- 1 Η,3H-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-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]-r-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]-l'-ethanol, N-(2-{[(2S)-3-(5-chloro-lΗ,3H-spiro[l- benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-5-chloro-4- hydroxyphenyl)acetamide, 2-Chloro-5- { [(2S)-3 -(5-chloro- 1 'H,3H-spiro[l -benzofuran-2,4'- piperidin] - 1 '-y l)-2-hydroxypropy ljoxy } -(4- { acety lamino } phenoxy)acetic acid, 5- { [(25)-3- (5-Chloro-r//,3H-spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-(4- { acety lamino }phenoxy)acetic acid, {2-Chloro-5-{[(25)-3-(5-chloro-rH,3H-spiro[l- benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy } -4-
[(methylamino)carbonyl]phenoxy } acetic acid, 2- {2-Chloro-5-{ [(25)-3 -(5-chloro- 1 '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 'H,3H-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy } -4- { [(3.S)-3- hydroxypyrrolidin-l-yl]carbonyl}phenoxy)acetic acid, 5-Chloro-2-{[(25)-3-(5-chloro- rH,3H-spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-4- (cyanomethoxy)benzoic acid, 2-{[(2S)-3-(5-chloro-lΗ,3Η-spiro[l-benzofuran-2,4'- piperidin]-r-yl)-2-hydroxypropyl]oxy}-5-chloro-4-(2,2-difluoroethoxy)benzoic, 5-Chloro- 2- { [(25)-3-(5-chloro- 1 'H,3H-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-yl)-2- hydroxypropy l]oxy } -4-(3 ,3 ,3 -trifluoropropoxy)benzoic acid, N-(2- { 3 -[5 -chloro- 1 Η,3 Η- spiro[l -benzofuran-2,4'-piperidin]- 1 '-yl]propoxy }phenyl)acetamide, Methyl 3-(2- { [(2S)-3- (5-chloro- 1 'H3H-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy}-4- fluoropheny l)propanoic acid, N-(2- { [(2S)-3-( { spiro [indole-2-4'-piperidin]-3 ( 1 Η)-one } - 1 '- yl)-2-hydroxypropyl]oxy}-4-hydroxyphenyl)acetamide, or (2-{[(2S)-3-(5-Chloro-l'H,3H- spiro[l-benzofuran-2,4'-piperidin]-l'-yl)-2-hydroxypropyl]oxy}-4- fluorophenyl)methanesulfonic acid, or a pharmaceutically acceptable salt thereof (for example as desscribed above; (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.
A chemokine antagonist (other than a CCRl antagonist), for example, 656933 (N-(2- bromophenyl)-N'-(4-cyano-lΗ-l,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.
5 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, OC 1768, OC2125, OC2184, OC499, OCD9101, I0 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- i5 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. 0
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. s 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, Rl 503, SC-80036, SCIO469, 6-chloro-5-[[(25,5/?)-4-[(4-fluorophenyl)methyl]- 2,5-domethyl- 1 -piperazinyl]carbonyl]-iV,N, 1 -trimethyl-α-oxo- 1 H-indole-3-acetamide,0 VX702 or VX745 (5-(2,6-dichlorophenyl)-2-(phenylthio)-6Η-pyrimido[ 1 ,6-b]pyridazin-6- one). A PDE Inhibitor: such as a PDE4 inhibitor, for example, 256066, Arofylline (3-(4- chlorophenyl)-3,7-dihydro-l -propyl- lH-Purine-2,6-dione), AWD 12-281 (N-(3,5-dichloro- 4-pyridinyl)-l-[(4-fluorophenyl)methyl]-5-hydroxy-α-oxo-lH-indole-3-acetamide), BAY19-8004 (Bayer), CDC-801 (Calgene), Celgene compound ((βR)-β-(3,4- dimethoxypheny I)- 1 ,3 -dihydro- 1 -oxo-2H-isoindole-2-propanamide), Cilomilast (cis-4- cyano-4-[3-(cyclopentyloxy)-4-methoxyphenyl]-cyclohexanecarboxylic acid), a compound in WO2006098353 from Kyowa Hakko Kogyo Co. Ltd. Japan, 2-(3,5-dichloro-4- pyridinyl)- 1 -(7-methoxyspiro[l ,3-benzodioxole-2, 1 '-cyclopentan]-4-yl)ethanone (CAS number 185406-34-2)), Compound from Pfizer (2-(3,4-difluorophenoxy)-5-fluoro-N-[cis- 4-[(2-hydroxy-5-methylbenzoyl)amino]cyclohexyl]-)-3-pyridinecarboxamide), Compound from Pfizer (2-(3, 4-difluorophenoxy)-5-fluoro-N-[cis-4-[[2-hydroxy-5- (hydroxymethyl)benzoyl]amino]cyclohexyl]-3-pyridinecarboxamide,), CT2820, GPD- 1116, Ibudilast, IC 485, KF 31334, KW-4490 (Kyowa Hakko Kogyo), Lirimilast ([2-(2,4- dichlorobenzoyl)-6-[(methylsulfonyl)oxy]-3-benzofuranyl])-urea), Merck Compound (N- cyclopropyl- 1 ,4-dihydro-4-oxo- 1 -[3-(3-pyridinylethynyl)phenyl]-)-l ,8-naphthyridine-3- carboxamide), Oglemilast (N-(3,5-dichloro-4-pyridinyl)-4-(difluoromethoxy)-8- [(methylsulfonyl)amino])-l-dibenzofurancarboxamide), ONO6126, ORG 20241 (4-(3,4- dimethoxyphenyl)-N-hydroxy-)-2-thiazolecarboximidamide), PD 189659/PD 168787 (Parke-Davis), Pentoxifylline (3,7-dihydro-3,7-dimethyl-l-(5-oxohexyl)-)-lH-purine-2,6- dione), Pfizer compound (5-fluoro-N-[4-[(2-hydroxy-4-methyl- benzoyl)amino]cyclohexyl]-2-(thian-4-yloxy)pyridine-3-carboxamide), Pfizer UK 500,001 , Piclamilast (3-(cyclopentyloxy)-N-(3,5-dichloro-4-pyridinyl)-4-methoxy- benzamide), PLX-369 (WO 2006026754), Roflumilast (3-(cyclopropylmethoxy)-N-(3,5- dichloro-4-pyridinyl)-4-(difluoromethoxy)benzamide), SCH 351591 (N-(3,5-dichloro-l- oxido-4-pyridinyl)-8-methoxy-2-(trifluoromethyl)-5-quinolinecarboxamide), SeICID(TM) CC- 10004 (Calgene), T-440 (Tanabe), Tetomilast (6-[2-(3,4-diethoxyphenyl)-4-thiazolyl]- 2-pyridinecarboxylic acid), Tofimilast (9-cyclopentyl-7-ethyl-6,9-dihydro-3-(2-thienyl)- 5H-pyrazolo[3,4-c]-l,2,4-triazolo[4,3-a]pyridine), TPI 1100, UCB 101333-3 (N,2- dicyclopropyl-6-(hexahydro- 1 H-azepin- 1 -yl)-5-methyl-4-pyrimidinamine), V- 11294 A (Napp), VM554A^M565 (Vernalis), or Zardaverine (6-[4-(difluoromethoxy)-3- methoxyphenyl]-3(2H)-pyridazinone); or a PDE5 Inhibitor, for example, Gamma- glutamyl[s-(2-iodobenzyl)cysteinyl]glycine, Tadalafϊl, Vardenafil, sildenafil, 4-phenyl- methylamino-6-chloro-2-(l-imidazolyl)-quinazoline, 4-phenyl-methylamino-6-chloro-2- (3-pyridyl)-quinazoline, l,3-dimethyl-6-(2-propoxy-5-methanesulphonylamidophenyl)- l,5-dihydropyrazolo[3,4-d]pyrimidin-4-one or l-cyclopentyl-3-ethyl-6-(3-ethoxy-4- pyridy l)-pyrazolo [3 ,4-d]pyrimidin-4-one.
For example, PDE4 inhibitors include 256066, Arofylline (3-(4-chlorophenyl)-3,7- dihydro-1 -propyl- lH-Purine-2,6-dione), AWD 12-281 (N-(3,5-dichloro-4-pyridinyl)-l-[(4- fluorophenyl)methyl]-5-hydroxy-α-oxo-lH-indole-3-acetamide), BAYl 9-8004 (Bayer), CDC-801 (Calgene), Celgene compound ((βR)-β-(3,4-dimethoxyphenyl)-l,3-dihydro-l- oxo-2H-isoindole-2-propanamide), Cilomilast (cis-4-cyano-4-[3-(cyclopentyloxy)-4- methoxyphenyl]-cyclohexanecarboxylic acid), a compound in WO2006098353 from Kyowa Hakko Kogyo Co. Ltd. Japan, 2-(3,5-dichloro-4-pyridinyl)-l-(7-methoxyspiro[l,3- benzodioxole-2,l'-cyclopentan]-4-yl)ethanone (CAS number 185406-34-2)), Compound from Pfizer (2-(3 ,4-difluorophenoxy)-5 -fluoro-N- [cis-4-[(2-hydroxy-5 - methylbenzoyl)amino]cyclohexyl]-)-3-pyridinecarboxamide), Compound from Pfizer (2- (3,4-difluorophenoxy)-5-fluoro-N-[cis-4-[[2-hydroxy-5-
(hydroxymethyl)benzoyl]amino]cyclohexyl]-3-pyridinecarboxamide,), CT2820, GPD- 1116, Ibudilast, IC 485, KF 31334, KW-4490 (Kyowa Hakko Kogyo), Lirimilast ([2-(2,4- dichlorobenzoyl)-6-[(methylsulfonyl)oxy]-3-benzofuranyl])-urea), Merck Compound (N- cyclopropyl- 1 ,4-dihydro-4-oxo-l -[3-(3-pyridinylethynyl)phenyl]-)- 1 ,8-naphthyridine-3- carboxamide), Oglemilast (N-(3 ,5-dichloro-4-pyridinyl)-4-(difluoromethoxy)-8- [(methylsulfonyl)amino])-l-dibenzofurancarboxamide), ONO6126, ORG 20241 (4-(3,4- dimethoxyphenyl)-N-hydroxy-)-2-thiazolecarboximidamide), PD 189659/PD 168787 (Parke-Davis), Pentoxifylline (3,7-dihydro-3,7-dimethyl-l -(5-oxohexyl)-)- 1 H-purine-2,6- dione), Pfizer compound (5-fluoro-N-[4-[(2-hydroxy-4-methyl- benzoyl)amino]cyclohexyl]-2-(thian-4-yloxy)pyridine-3-carboxamide), Pfizer UK 500,001, Piclamilast (3-(cyclopentyloxy)-N-(3,5-dichloro-4-pyridinyl)-4-methoxy- benzamide), PLX-369 (WO 2006026754), Roflumilast (3-(cyclopropylmethoxy)-N-(3,5- dichloro-4-pyridinyl)-4-(difluoromethoxy)benzamide), SCH 351591 (N-(3,5-dichloro-l- oxido-4-pyridinyl)-8-methoxy-2-(trifluoromethyl)-5-quinolinecarboxamide), SeICID(TM) CC- 10004 (Calgene), T-440 (Tanabe), Tetomilast (6-[2-(3,4-diethoxyphenyl)-4-thiazolyl]- 2-pyridinecarboxylic acid), Tofimilast (9-cyclopentyl-7-ethyl-6,9-dihydro-3-(2-thienyl)- 5H-pyrazolo[3,4-c]-l,2,4-triazolo[4,3-a]pyridine), TPI 1100, UCB 101333-3 (N,2- dicyclopropyl-6-(hexahydro- 1 H-azepin- 1 -yl)-5-methyl-4-pyrimidinamine), V- 11294A (Napp), VM554/VM565 (Vernalis), or Zardaverine (6-[4-(difluoromethoxy)-3- methoxyphenyl]-3(2H)-pyridazinone. For example, a PDE4 inhibitor is Roflumilast ((3-(cyclopropylmethoxy)-N-(3,5-dichloro- 4-pyridinyl)-4-(difluoromethoxy)benzamide).
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 Doxycycline).
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 active ingredients may be in the form of a solvate, e.g. a hydrate.
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 7-[(l/?)-2-({2-[(3-{[2-(2-
Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient 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 which is Aclidinium bromide, Glycopyrrolate (such as R5R-, R,S-, S,R-, or S,S-glycopyrronium bromide), Oxitropium bromide, Pirenzepine, telenzepine or
Tiotropium bromide; a p38 inhibitor; or, a PDE inhibitor.
In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl]amino}propyl)thio]ethyl}afnino)- 1 -hydroxyethyl]-4-hydroxy- 1 ,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient which is a nonsteroidal Glucocorticoid Receptor (GR) Agonist for example, a compound disclosed in WO 2006/046916. In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient which is a CCRl antagonist, 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)-l-[[2- [(aminocarbonyl)amino]-4-chlorophenoxy]acetyl]-4-[(4-fluorophenyl)methyl]-2- methylpiperazine monohydrochloride) or CCX634.
In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient which is a chemokine antagonist (not CCRl), for example, 656933 (N-(2-bromophenyl)-N'-(4-cyano- 1 Η- 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- { 1 - [3 -(4-ethoxyphenyl)-4-oxo-3 ,4-dihydropyrido [2,3 -d]pyrimidin-2-y l]ethyl } -N- (pyridin-3-ylmethyl)-2-[4-(trifluoromethoxy)phenyl]acetamide) or Vicriviroc.
In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(lΛ)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a corticosteroid, 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. In one embodiment of the present invention the corticosteroid is selected from budesonide, fluticasone propionate, fluticasone fruoate mometasone furoate, beclomethasone dipropionate or butixocort propionate ester.
In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(li?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a corticosteroid, for example, Budesonide, Fluticasone Furoate or Fluticasone propionate.
In one embodiment of the present invention the corticosteroid is budesonide. Budesonide and its preparation is described, for example, in Arzneimittel-Forschung (1979), 29 (11), 1687-1690, DE 2,323,215 and US 3,929,768. Presently available formulations of budesonide are marketed under the tradename 'Entocort ®'.
In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(li?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is an IKK2 inhibitor, for example, 2-{[2-(2-Methylamino-pyrimidin-4-yl)-lΗ-indole-5-carbonyl]- amino}-3-(phenyl-pyridin-2-yl-amino)-propionic acid.
In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(li?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)- 1 -hydroxyethyl]-4-hydroxy- 1 ,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a muscarinic antagonist, for example, Aclidinium bromide, Glycόpyrrolate (such as R5R-, R,S-, S5R-, or S,S-glycopyrronium bromide), Oxitropium bromide, Pirenzepine, telenzepine or Tiotropium bromide.
In one aspect of the invention the muscarinic receptor antagonist is a long acting muscarinic receptor antagonist, i.e. 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 7-[(li?)-2-({2-[(3-{[2-(2-
Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is Tiotropium bromide.
In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)- 1 -hydroxyethyl]-4-hydroxy- 1 ,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a p38 inhibitor, 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(3Η)- pyrimidinone), Array-797, AZD6703, Doramapimod, KC-706, PH 797804, Rl 503, SC- 80036, SCIO469, 6-chloro-5-[[(25,5/?)-4-[(4-fluorophenyl)methyl]-2,5-domethyl-l- piperaziny^carbony^-iV^l-trimethyl-α-oxo-lH-indole-S-acetamide, VX702 or VX745 (5-(2,6-dichlorophenyl)-2-(phenylthio)-6Η-pyrimido[l,6-b]pyridazin-6-one).
In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(li?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a PDE Inhibitor: such as a PDE4 inhibitor {for example, 256066, Arofylline (3-(4-chlorophenyl)- 3,7-dihydro-l-propyl-lΗ-Purine-2,6-dione), AWD 12-281 (N-(3,5-dichloro-4-pyridinyl)- 1 -[(4-fluorophenyl)methyl]-5-hydroxy-α-oxo- 1 H-indole-3-acetamide), BAYl 9-8004 (Bayer), CDC-801 (Calgene), Celgene compound ((βR)-β-(3,4-dimethoxyphenyl)-l,3- dihydro- 1 -oxo-2H-isoindole-2-propanamide), Cilomilast (cis-4-cyano-4-[3- (cyclopentyloxy)-4-methoxyphenyl]-cyclohexanecarboxylic acid), a compound in
WO2006098353 from Kyowa Hakko Kogyo Co. Ltd. Japan, 2-(3,5-dichloro-4-pyridinyl)- 1 -(7-methoxyspiro[ 1 ,3-benzodioxole-2, 1 '-cyclopentan]-4-yl)ethanone (CAS number 185406-34-2)), Compound from Pfizer (2-(3,4-difluorophenoxy)-5-fluoro-N-[cis-4-[(2- hydroxy-5-methylbenzoyl)amino]cyclohexyl]-)-3-pyridinecarboxamide), Compound from Pfizer (2-(3,4-difluorophenoxy)-5-fluoro-N-[cis-4-[[2-hydroxy-5- (hydroxymethyl)benzoyl]amino]cyclohexyl]-3-pyridinecarboxamide,), CT2820, GPD- 1116, Ibudilast, IC 485, KF 31334, KW-4490 (Kyowa Hakko Kogyo), Lirimilast ([2-(2,4- dichlorobenzoyl)-6-[(methylsulfonyl)oxy]-3-benzofuranyl])-urea), Merck Compound (N- cyclopropyl- 1 ,4-dihydro-4-oxo-l -[3-(3-pyridinylethynyl)phenyl]-)- 1 ,8-naphthyridine-3- carboxamide), Oglemilast (N-(3,5-dichloro-4-pyridinyl)-4-(difluoromethoxy)-8- [(methylsulfonyl)amino])-l-dibenzofurancarboxamide), ONO6126, ORG 20241 (4-(3,4- dimethoxyphenyl)-N-hydroxy-)-2-thiazolecarboximidamide), PDl 89659/PD 168787
(Parke-Davis), Pentoxifylline (3,7-dihydro-3,7-dimethyl-l-(5-oxohexyl)-)-lH-purine-2,6- dione), Pfizer compound (5-fiuoro-N-[4-[(2-hydroxy-4-methyl- benzoyl)amino]cyclohexyl]-2-(thian-4-yloxy)pyridine-3-carboxamide), Pfizer UK 500,001 , Piclamilast (3-(cyclopentyloxy)-N-(3,5-dichloro-4-pyridinyl)-4-methoxy- benzamide), PLX-369 (WO 2006026754), Roflumilast (3-(cyclopropylmethoxy)-N-(3,5- dichloro-4-pyridinyl)-4-(difluoromethoxy)benzamide), SCH 351591 (N-(3,5-dichloro-l- oxido-4-pyridinyl)-8-methoxy-2-(trifluoromethyl)-5-quinolinecarboxamide), SeICID(TM) CC- 10004 (Calgene), T-440 (Tanabe), Tetomilast (6-[2-(3,4-diethoxyphenyl)-4-thiazolyl]- 2-pyridinecarboxylic acid), Tofimilast (9-cyclopentyl-7-ethyl-6,9-dihydro-3-(2-thienyl)- 5H-pyrazolo[3,4-c]-l,2,4-triazolo[4,3-a]pyridine), TPI 1100, UCB 101333-3 (N,2- dicyclopropyl-6-(hexahydro- 1 H-azepin- 1 -yl)-5 -methyl-4-pyrimidinamine), V-I l 294 A (Napp), VM554/VM565 (Vernalis), or Zardaverine (6-[4-(difluoromethoxy)-3- methoxyphenyl]-3(2H)-pyridazinone); or a PDE5 Inhibitor, for example, Gamma- glutamyl[s-(2-iodobenzyl)cysteinyl]glycine, Tadalafil, Vardenafil, sildenafil, 4-phenyl- methy lamino-6-chloro-2-( 1 -imidazolyl)-quinazoline, 4-phenyl-methy lamino-6-chloro-2- (3-pyridyl)-quinazoline, l,3-dimethyl-6-(2-propoxy-5-methanesulphonylamidophenyl)- 1 ,5-dihydropyrazolo[3,4-d]pyrimidin-4-one or 1 -cyclopentyl-3-ethyl-6-(3-ethoxy-4- pyridyl)-pyrazolo[3,4-d]pyrimidin-4-one}.
In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a PDE4 inhibitor, for example, 256066, Arofylline (3-(4-chlorophenyl)-3,7-dihydro-l -propyl- 1Η- Purine-2,6-dione), AWD 12-281 (N-(3,5-dichloro-4-pyridinyl)-l-[(4- fluorophenyl)methyl]-5-hydroxy-α-oxo- 1 Η-indole-3-acetamide), BAY 19-8004 (Bayer), CDC-801 (Calgene), Celgene compound ((βR)-β-(3,4-dimethoxyphenyl)- 1 ,3-dihydro- 1 - oxo-2H-isoindole-2-propanamide), Cilomilast (cis-4-cyano-4- [3 -(cyclopenty loxy)-4- methoxyphenyl]-cyclohexanecarboxylic acid), a compound in WO2006098353 from Kyowa Hakko Kogyo Co. Ltd. Japan, 2-(3,5-dichloro-4-pyridinyl)-l-(7-methoxyspiro[l,3- benzodioxole-2,l'-cyclopentan]-4-yl)ethanone (CAS number 185406-34-2)), Compound from Pfizer (2-(3,4-difluorophenoxy)-5-fluoro-N-[cis-4-[(2-hydroxy-5- methylbenzoyl)amino]cyclohexyl]-)-3-pyridinecarboxamide), Compound from Pfizer (2- (3,4-difluorophenoxy)-5-fluoro-N-[cis-4-[[2-hydroxy-5-
(hydroxymethy^benzoylJaminoJcyclohexylJ-S-pyridinecarboxamide,), CT2820, GPD- 1116, Ibudilast, IC 485, KF 31334, KW-4490 (Kyowa Hakko Kogyo), Lirimilast ([2-(2,4- dichlorobenzoyl)-6-[(methylsulfonyl)oxy]-3-benzofuranyl])-urea), Merck Compound (N- cyclopropyl- 1 ,4-dihydro-4-oxo- 1 -[3-(3-pyridinylethynyl)phenyl]-)- 1 ,8-naphthyridine-3- carboxamide), Oglemilast (N-(3,5-dichloro-4-pyridinyl)-4-(difluoromethoxy)-8- [(methylsulfonyl)amino])-l-dibenzofurancarboxamide), ONO6126, ORG 20241 (4-(3,4- dimethoxyphenyl)-N-hydroxy-)-2-thiazolecarboximidamide), PD189659/PD168787 (Parke-Davis), Pentoxifylline (3,7-dihydro-3,7-dimethyl-l-(5-oxohexyl)-)-lH-purine-2,6- dione), Pfizer compound (5-fluoro-N-[4-[(2-hydroxy-4-methyl- benzoyl)amino]cyclohexyl]-2-(thian-4-yloxy)pyridine-3-carboxamide), Pfizer UK 500,001 , Piclamilast (3-(cyclopentyloxy)-N-(3,5-dichloro-4-pyridinyl)-4-methoxy- benzamide), PLX-369 (WO 2006026754), Roflumilast (3-(cyclopropylmethoxy)-N-(3,5- dichloro-4-pyridinyl)-4-(difluoromethoxy)benzamide), SCH 351591 (N-(3,5-dichloro-l- oxido-4-pyridinyl)-8-methoxy-2-(trifluoromethyl)-5-quinolinecarboxamide), SeICID(TM) CC- 10004 (Calgene), T-440 (Tanabe), Tetomilast (6-[2-(3,4-diethoxyphenyl)-4-thiazolyl]- 2-pyridinecarboxylic acid), Tofimilast (9-cyclopentyl-7-ethyl-6,9-dihydro-3-(2-thienyl)- 5H-pyrazolo[3,4-c]-l,2,4-triazolo[4,3-a]pyridine), TPI 1100, UCB 101333-3 (N,2- dicyclopropyl-6-(hexahydro-lH-azepin-l-yl)-5-methyl-4-pyrimidinamine), V-11294 A (Napp), VM554/VM565 (Vernalis), or Zardaverine (6-[4-(difluoromethoxy)-3- methoxyphenyl]-3(2H)-pyridazinone). In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(l/?)-2-({2-[(3-{[2-(2- Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is a PDE4 5 inhibitor, for example AWD 12-281 (N-(3,5-dichloro-4-pyridinyl)-l-[(4- fluoropheny l)methyl]-5-hydroxy-α-oxo- 1 Η-indole-3-acetamide) or roflumilast.
In another aspect the present invention provides a pharmaceutical product comprising, in combination, a first active ingredient which is 7-[(lΛ)-2-({2-[(3-{[2-(2- i o Chlorophenyl)ethy l]amino } propyl)thio]ethyl } amino)- 1 -hydroxyethyl]-4-hydroxy- 1,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient is roflumilast.
In another aspect the present invention provides a kit comprising a preparation of a first active ingredient which is 7-[(li?)-2-({2-[(3-{[2-(2- i5 Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one or a salt thereof, a preparation of a second active ingredient selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; an antioxidant; 20 a CCRl antagonist; a chemokine antagonist (not CCRl); a corticosteroid; a CRTh2 antagonist; a DPI antagonist; 5 an Ηistone Deacetylase Inducer; an IKK2 inhibitor; a COX inhibitor; a lipoxygenase inhibitor; a leukotriene receptor antagonist; 30 an MPO inhibitor; a muscarinic antagonist which is Aclidinium bromide, Glycopyrrolate (such as R5R-, R,S-,
S,R-, or S,S-glycopyrronium bromide), Oxitropium bromide, Pirenzepine, telenzepine or
Tiotropium bromide; a p38 inhibitor; a PDE 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.
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 simultaneous it 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 e.g. 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 colourant. 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, they are in admixture). Therefore, the present invention further provides a pharmaceutical composition comprising, in admixture, a first active ingredient which is 7-[(li?)-2-({2-[(3-{[2-(2-
Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)- 1 -hydroxyethyl]-4-hydroxy- 1 ,3- benzothiazol-2(3H)-one Dihydrobromide, and a second active ingredient as defined above. The pharmaceutical composition 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 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 first and second 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 is a hydrocarbon, chlorofluorocarbon or hydrofluoroalkane (e.g. heptafluoroalkane) propellant, or mixture of any such propellant, for example in a pressurised metered dose inhaler (pMDI). A preferred propellant is P 134a or P227, each of which may be used alone or in combination with other propellants and/or surfactant and/or other excipients. 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.
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).
Dry powder inhalers may be used to administer the active ingredients, alone or in combination with a pharmaceutically acceptable carrier, in the later case either as a finely divided powder or as an ordered mixture. The dry powder inhaler may be single dose or multi-dose and may utilise a dry powder or a powder-containing capsule.
Metered dose inhaler, nebuliser and dry powder inhaler devices are well known and a variety of such devices are available.
The pharmaceutical product 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 NSAID-induced) and dust-induced asthma, both intermittent and persistent and of all severities, and other causes of airway hyper-responsiveness; chronic obstructive pulmonary disease (COPD); bronchitis, including infectious and eosinophilic bronchitis and chronic 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 s 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 I0 fever); nasal polyposis; acute viral infection including the common cold, and infection due to respiratory syncytial virus, influenza, coronavirus (including SARS) and adenovirus.
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 is particular chronic obstructive pulmonary disease or asthma.
The present invention still further provides a method of treating a respiratory disease which comprises simultaneously, sequentially or separately administering: (a) a therapeutically effective dose of a first active ingredient as defined above; and,0 (b) a therapeutically effective dose of a second active ingredient as defined above; to a patient in need thereof.
In the context of the present specification, the term "therapy" also includes "prophylaxis" unless there are specific indications to the contrary. The terms "therapeutic" and 5 "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 by0 genetic testing or screening to be particularly susceptible to developing the condition or disorder. General Preparative Methods
There follow preparative methods for 7-[(li?)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]- amino } propy l)thio]ethy 1 } amino)- 1 -hydroxyethyl] -4-hydroxy- 1 ,3 -benzothiazol-2(3H)-one Dihydrobromide and also assays and data showing the activity of this compound (called Compound A in the assays and Table 1 below).
1H NMR spectra were recorded on a Varian Inova 400 MHz or a Varian Mercury-WX 300 MHz instrument. The central peaks of chloroform-^ (5H 7.27 ppm), dimethylsulfoxide-c4 (5H 2.50 ppm), acetonitrile-c/j (5H 1-95 ppm) or methanol-^ (5H 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 C18-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 examples have the following meanings: SCX: Solid phase extraction with a sulfonic acid sorbent HPLC: High performance liquid chromatography DMF: N,iV-Dimethylformamide
Figure 1 Shows the onset times for roflumilast (lμM), Compound A (0.InM) and Compound A (0.InM) in the presence of roflumilast (lμM) in guinea pig trachea in vitro. Preparation 1
7-KlR)-2-f{2-f(3-{[2-(2-Chlorophenvnethyllamino}propyl)thiolethyl}aniino)-l- hvdroxyethyll-4-hydroxy-l,3-benzothiazol-2(3H)-one dihydrobromide (Compound A)
Figure imgf000027_0001
a) l-Chloro-2-[(£)-2-nitrovinyl]benzene
Figure imgf000027_0002
2-Chlorobenzaldehyde (ex Aldrich) (10.0 g) was mixed with nitromethane (26.05 g) and ammonium acetate (21.92 g) in acetic acid (200 mL), and the mixture was heated at reflux for 40 minutes. The mixture was allowed to cool to room temperature, and the majority of the acetic acid was removed in vacuo. The residue was dissolved in dichloromethane and washed with water, then potassium carbonate solution (x2), then water again. The organics were dried over anhydrous magnesium sulfate, filtered and evaporated to give the desired material, as an orange oil (12.83 g). 1H NMR δ( CDCl3) 8.41 (d, IH), 7.62-7.57 (m, 2H), 7.52-7.48 (m, IH), 7.43 (dt, IH), 7.34 (ddd, IH)
b) 2-(2-Chlorophenyl)ethanamine
Figure imgf000027_0003
Aluminium hydride was prepared by the drop- wise addition of a solution of sulphuric acid (8.40 mL) in dry THF (60 mL) to a stirred solution of 1.0M lithium aluminium hydride in THF (314 mL), at 0-100C, under a nitrogen atmosphere. After stirring at 50C for 30 minutes, a solution of l-chloro-2-[(£)-2-nitrovinyl]benzene (12.83 g) in dry THF (160 mL) was added dropwise maintaining the internal temperature between O0C and 1O0C. When the addition was complete the reaction was heated at reflux for 5 minutes. The mixture was allowed to cool to room temperature, then cooled to O0C and isopropanol (22 mL) carefully added dropwise maintaining the temperature below 2O0C. 2M Sodium hydroxide (35 mL) was carefully added dropwise maintaining the temperature below 2O0C. The mixture was stirred at room temperature for 30 minutes, then filtered through a layer of celite, which was then washed with THF (x3). The filtrate was evaporated to dryness. The residue was purified using silica column chromatography, using ethyl acetate to load the material, then 10% triethylamine in ethyl acetate, followed by 10% triethylamine in 45% ethanol: 45% ethyl acetate as the eluents, to give the desired material (4.66 g). 1H NMR δ( CDCl3) 7.36 (dd, IH), 7.25-7.13 (m, 3H), 2.98 (dt, 2H), 2.91-2.87 (m, 2H)
c) ter/-Butyl [2-(2-chlorophenyl)ethyl]carbamate
Figure imgf000028_0001
To a stirred solution of 2-(2-chlorophenyl)ethanamine (25.57 g) and triethylamine (22.87 mL) in dry THF (300 mL) was added a solution of di-tert-butyl dicarbonate (35.85 g) in dry THF (50 mL) over 10 minutes, at ambient temperature, under a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 3 hours. The solvents were removed in vacuo to give the desired material, as a yellow oil (42.0 g). 1H NMR δ(CDCL3) 7.35 (d, IH), 7.25-7.14 (m, 3H), 4.57 (s, IH), 3.43-3.35 (m, 2H), 2.95 (t, 2H), 1.43 (d, 9H)
d) tert-Butyl allyl[2-(2-chlorophenyl)ethyl] carbamate
Figure imgf000028_0002
To a suspension of sodium hydride (60% in mineral oil) (7.23 g), which had been washed with ether (x3), in dry DMF (200 mL) was added a solution of tert-butyl [2-(2- chlorophenyl)ethyl]carbamate (42.0 g) in dry DMF (50 mL), over a 15 minute period, at 350C, under a nitrogen atmosphere. When the addition was complete, the mixture was stirred at 5O0C for 90 minutes. The mixture was allowed to cool to room temperature, then allyl bromide (15.63 mL) was added slowly, keeping the temperature at 250C, using external cooling. The mixture was stirred at room temperature for 2 hours, then diluted with water and extracted with ethyl acetate (x3). The organics were combined, washed with water, dried over anhydrous magnesium sulfate, filtered and evaporated. The residue
5 was purified using silica column chromatography, loading with 1% ethyl acetate in isohexane, then using isohexane with ethyl acetate (0%, 1%, 2%, %5) as the eluents to give the desired material (27.0 g). There were several mixed fractions, so these were combined, and re-purified using silica column chromatography, as above, to give a further 4g of desired material. Both crops of product were combined to give 31.0 g in total. io 1H NMR 5( CDCl3) 7.36-7.31 (m, IH), 7.21-7.12 (m, 3H), 5.83-5.68 (m, IH), 5.17-5.05 (m, 2H), 3.86-3.66 (m, 2H), 3.41 (t, 2H), 3.03-2.90 (m, 2H), 1.43 (s, 9H) HPLC: 95.90% @ 220nm [M+H-Boc]+ = 196.1 (CaIc = 295.1339) (multimode+)
e) tert-Butyl [2-(2-chlorophenyl)ethyl] {3-[(2-hydroxyethyl)thio]propyl}carbamate
Figure imgf000029_0001
tert-Butyl allyl[2-(2-chlorophenyl)ethyl]carbamate (31.0 g) was mixed with 2-mercaptoethanol (7.37 mL), and AIBN (1.15 g), and stirred at 650C for 45 minutes. The mixture was cooled and more mercaptoethanol (1 mL) and AIBN (200 mg) added. The mixture was then heated at 650C for a further 30 minutes. The material was purified by 0 silica column chromatography, loading the material in 20% ethyl acetate in isohexane, then eluting with 20% ethyl acetate in isohexane, changing to 50%, to give the desired material (31.94 g).
1H NMR δ( CDCl3) 7.38-7.32 (m, IH), 7.22-7.13 (m, 3H), 3.75-3.68 (m, 2H), 3.41 (t, 2H), 3.32-3.14 (m, 2H), 3.03-2.91 (m, 2H), 2.72 (t, 2H), 2.54-2.36 (m, 2H), 1.85-1.71 (m, 2H),
2S 1.42 (s, 9H)
HPLC: 92.31% @ 220nm [M+H-Boc]+ = 274.1 (CaIc = 373.1478) (multimode+)
f) tert-Butyl [2-(2-chlorophenyl)ethyl]{3-[(2-oxoethyl)thio]propyl}carbamate
Figure imgf000030_0001
Sulfur trioxide:pyridine complex (30.52 g) was dissolved in DMSO (200 mL) and stirred at room temperature, under a nitrogen atmosphere, for 15 minutes. DCM (100 mL) was added, followed by a solution of tert-butyl [2-(2-chlorophenyl)ethyl]{3-[(2- hydroxyethyl)thio]propyl}carbamate(23.9 g) and Hunigs base (63.5 mL) in DCM (160 mL), which was added in one portion (exotherm). The resulting mixture was stirred at ambient temperature for 15 minutes. The reaction mixture was diluted with ethyl acetate, washed with water, then IN HCl, then saturated sodium bicarbonate solution, dried over anhydrous magnesium sulfate, filtered and the solvents removed in vacuo. The material was purified by silica column chromatography eluting with 20% ethyl acetate in isohexane to give the desired material (12.43 g).
1H NMR δ( CDCl3) 9.46 (t, IH), 7.36-7.32 (m, IH), 7.21-7.13 (m, 3H), 3.40 (t, 2H), 3.29- 3.13 (m, 4H), 3.02-2.90 (m, 2H), 2.45-2.34 (m, 2H), 1.82-1.69 (m, 2H), 1.49-1.36 (m, 9H)
g) tert-Buty\ [2-(2-chlorophenyl)ethyl] {3-[(2-{[(2/?)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3- dihydro-l,3-benzothiazol-7-yI)ethyl]amino}ethyl)thio|propyl}carbamate
Figure imgf000030_0002
The tert-butyl [2-(2-chlorophenyl)ethyl]{3-[(2-oxoethyl)thio]propyl}carbamate (11.32 g) was dissolved in a mixture of methanol (200 mL) and acetic acid (1.74 ml). 7-[(l/?)-2- amino- l-hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3H)-one hydrochloride (8.0 g) was added to the solution, and the mixture stirred at room temperature, under a nitrogen atmosphere, for 1 hour. Sodium cyanoborohydride (1.92 g) was added and the mixture stirred for a further 2 hours. The solvents were removed in vacuo, and the residue diluted with water, basified with 0.880 aqueous ammonia, and extracted with ethyl acetate (x3) (filtered through celite during extraction). The organics were combined, washed with brine, dried over anhydrous sodium sulfate, filtered and evaporated to give a brown residue (15.5 g). The material was purified using silica column chromatography, using DCM with MeOH (2%, 5%, 10%, 20% and 30%, all with 1% 0.880 aq NH3) as the eluent, to give the desired material (6.67 g) (38% yield)
1H NMR δ(DMSO) 7.43-7.38 (m, IH), 7.30-7.21 (m, 3H), 6.86 (d, IH), 6.69 (d, IH), 4.56 (dd, IH), 3.23-3.10 (m, 2H), 2.88 (t, 2H), 2.71-2.48 (m, 8H), 2.46-2.39 (m, 2H), 1.72-1.62 (m, 2H), 1.40-1.22 (m, 9H) HPLC: 97.46% @ 220nm [M+H]+=582.1 (CaIc = 582.1863) (multimode+)
h) 7-[(lR)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l- hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3//)-one dihydrobromide
Figure imgf000031_0001
To a stirred suspension of the Boc compound from part g) (5.93 g) in DCM (20 mL) was added trifluoroacetic acid (20 mL) at O0C, and the resulting mixture was stirred under nitrogen for 30 minutes. The mixture was diluted with toluene, and solvents removed, then azeotroped with toluene (x2). The residue was dissolved in acetonitrile, acidified with 48% aq HBr and concentrated in vacuo (not to dryness). The mixture was further diluted with acetonitrile and the precipitated solid collected by filtration, washed with acetonitrile and dried under vacuum to give 6.35 g. A 3.8% impurity was present (isomer from part e)), so the material was redissolved in a 1:1 mixture of acetonitrile: water and purified using prep HPLC (Sunfire 30x80mm C8 column; NH4OAc buffer; acetonitrile 5-50% over 10 minutes). The resultant material was dried overnight in a dessicator at 10 mbar over KOH and H2SO4. The resulting di-acetate salt was dissolved in water and basified with 0.880 aq ammonia. A white gum formed, so the aqueous was decanted off, and the gum dried in vacuo to give the free base (4.11 g). This was dissolved in hot ethanol, and the solution was filtered, then allowed to cool to room temperature. The solution was acidified with 48% aq. HBr and left to crystallize. The white solid was collected by filtration, washed with ethanol and dried in vacuo to give 3.81 g Crop 1. 'H NMR δ(DMSO) 11.67 (s, IH), 10.15 (s, IH), 8.70 (s, 4H), 7.50-7.30 (m, 4H), 6.94 (d, IH), 6.78 (d, IH), 6.45 (s, IH), 4.96-4.90 (m, IH), 3.22-3.02 (m, 10H), 2.86-2.76 (m, 2H), 2.66 (t, 2H), 1.91 (quintet, 2H) HPLC: 99.63% @ 220nm [M+H]+=482 (calc=482.1339) (MultiMode+)
5
Elemental analysis: C H N S Calculated: 41.04 4.70 6.53 9.96 Found: 1 : 41.07 4.69 6.67 9.72 2: 41.08 4.68 6.74 9.67
I0 3: 40.96 4.68 6.75 9.67
The mother liquors were evaporated to dryness then triturated with acetonitrile. The solid was collected by filtration to give 719 mg Crop 2 (4.53 g total). 1H NMR δ(DMSO) 11.67 (s, IH), 10.15 (s, IH), 8.80-8.60 (m, 4H), 7.50-7.29 (m, 4H), 6.94 (d, IH), 6.78 (d, IH), 6.45 (s, IH), 4.96-4.89 (m, IH), 3.22-3.00 (m, 10H), 2.85-2.76 is (m, 2H), 2.66 (t, 2H), 1.90 (quintet, 2H)
HPLC: 99.20% @ 220nm [M+H]+=482 (calc=482.1339) (MultiMode+) Elemental analysis: C H N S Calculated: 41.04 4.70 6.53 9.96
Found: 1: 40.90 4.69 6.78 9.60
20 2: 41.01 4.70 6.83 9.60
3: 40.97 4.69 6.76 9.63
Alternative salts of 7-[( 1 R)-2-( { 2- [(3 - { [2-(2-Chloropheny l)ethyl]amino } propy l)thio] - ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3H)-one can be prepared by 25 mixing a suitable acid with 7-[(l/?)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)- thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3H)-one in a solvent. Examples are provided below. Preparation 2
7-[(l/?)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l- hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3//)-one Dihydrobromide - Polymorph B
a) iV-[2-(2-Chlorophenyl)ethyl]acrylamide
Figure imgf000033_0001
Hunigs base
Figure imgf000033_0002
Figure imgf000033_0003
93%
2-(2-Chlorophenyl)ethanamine (ex Aldrich) (leq, 49 Ig, 3.16mol) was dissolved in dichloromethane (2500ml). The solution was cooled to O0C and Hunigs base (leq, 522ml, 3.16mol) added. Aery loyl chloride ( 1 eq, 257ml, 3.16mol) was added dropwise, keeping the temperature between O0C and 50C throughout the 2 hour addition. The reaction was warmed to ambient temperature and stirred overnight. The mixture was diluted with dichloromethane (1500ml) and washed with 2M HCl (2 x 1000ml), then water (1 x 1000ml), dried over anhydrous sodium sulfate, filtered and evaporated to give the desired material as a white, waxy solid (646g) (97% yield).
1H NMR δ(CDci3) 3.01 (t, J= 7.1 Hz, 2H), 3.61 (q, J= 6.8 Hz, 2H), 5.62 (d, J= 10.6 Hz, 2H), 6.08 (dd, J= 17.6, 10.6 Hz, IH), 6.25 (d, J= 17.6 Hz, IH), 7.18-7.24 (m, 3H), 7.34- 7.38 (m, IH)
HPLC: 94.02% @ 220nm [M+H]+=210.1 (calc=210.0685) (MultiMode+)
b) [(3-{[2-(2-Chlorophenyl)ethyl]amino}-3-oxopropyl)thio]acetic acid
Figure imgf000033_0004
100% Ethyl mercaptoacetate (leq, 138ml, 1.25mol) was dissolved in ethanol (750ml) and sodium ethoxide (21% weight in ethanol) (leq, 405ml, 1.25mol) added, keeping the internal temperature below 3O0C throughout. The reaction mixture was stirred for 1 hour before a solution of N-[2-(2-chlorophenyl)ethyl]acrylamide (leq, 261.8g, 1.25mol) in ethanol (2250ml) was added dropwise (no increase in temperature was noted). The mixture was stirred for 18 hours. A further batch of ethyl mercaptoacetate (0.15eq) and sodium ethoxide (0.15eq) were added, and mixture stirred for a further 24 hours. A further aliquot of sodium ethoxide (20ml) was added, followed by the slow addition of water (1000ml), keeping the temperature below 2O0C. The mixture was then stirred for 24 hours at ambient temperature. LC-MS showed complete conversion to the acid. The mixture was concentrated in vacuo to a volume of ~1 litre, and another litre of water added to the mixture. The mixture was washed with tert-butyl methyl ether. The aqueous layer was acidified to pH 1 with cone. HCl, then extracted with /ert-butyl methyl ether (Ix 21itre, Ix 1.51itre). The organics were combined, washed with water (Ix llitre), dried over anhydrous sodium sulfate, filtered and evaporated to give the desired material as a yellow oil (357.9Ig) (95% yield).
1H NMR δ(cDCi3) 2.51 (t, J= 7.6 Hz, 2H), 2.93 (t, J= 7.2 Hz, 2H), 2.97 (t, J= 7.2 Hz, 2H), 3.26 (s, 2H), 3.55 (q, J= 6.4 Hz, 2H), 6.12 (s, IH), 7.15-7.25 (m, 3H), 7.33-7.36 (m, IH)
HPLC: 86.02% @ 220nm [M+H]+=302.1 (calc=302.0617) (MultiMode+)
c) 2- [(3-{ [2-(2-Chlorophenyl)ethyl] amino} propy l)thio] ethanol
Figure imgf000034_0001
[(3-{[2-(2-Chlorophenyl)ethyl]amino}-3-oxopropyl)thio]acetic acid (107.7g, 357mmol) was dissolved in THF (llitre), and a IM borane in THF solution (ex Aldrich) (1.5 litre) was added drop-wise over ~4 hours. The internal temperature was maintained at 3O0C ±5°C throughout the addition. The reaction was then heated to 650C (internal temperature) overnight, with stirring. Methanol (500ml) was added drop-wise, followed by 2M HCl (500ml), and the reaction re fluxed gently for 4 hours. The reaction was cooled, concentrated in vacuo to a volume of ~1 litre, and a litre of water was added to the mixture. This mixture was washed with tert-butyl methyl ether (2x 500ml). The aqueous layer was basified to ~pH 9 with solid sodium hydroxide, then extracted with tert-butyl methyl ether (3 x 500ml). The organics were combined, washed with water (1 x 500ml), dried over anhydrous sodium sulfate, filtered and evaporated to give the desired material (85.3g) (87% yield).
1H NMR δ(CDci3) 1.73-1.82 (quintet, 2H), 1.96 (s, IH), 2.63 (t, J= 7.2 Hz, 2H), 2.72 (t, J = 5.8 Hz, 2H), 2.78 (t, J= 6.8 Hz, 2H), 2.83-2.97 (m, 4H), 3.74 (t, J= 5.9 Hz, 2H), 7.12-7.25 (m, 3H), 7.33-7.36 (m, IH)
HPLC: 89.70% @ 220nm [M+H]+=274.1 (calc=274.1032) (MultiMode+)
d) tert-Butyl [l^l-chloropheny^ethylJIS-^-hydroxyethyOthioJpropylJcarbamate
Figure imgf000035_0001
2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thiό]ethanol (leq, 85.0g, 312mmol) was dissolved in dichloromethane (600ml) and cooled in an ice bath. Hunigs base (leq, 51.5ml, 312mmol) was added, followed by a solution of di-tert-butyl dicarbonate (leq, 68.1g, 312mmol) in dichloromethane (250ml), which was added dropwise over ~2 hours to maintain an internal temperature of- 50C. The cooling bath was removed, and the reaction allowed to stir overnight, warming to ambient temperature. Dichloromethane (500ml) was added and the reaction mixture was washed with water (2x 500 ml), 2M HCl (2x 500ml), then water (2x 500ml) again, before being dried over anhydrous sodium sulfate, filtered and evaporated to give the desired material, as a pale, yellow oil (116g) (100% yield). 1H NMR δ(cDCi3) 1-42 (s, 9H), 1.74-1.84 (m, 2H), 2.46-2.53 (m, 2H), 2.72 (t, J= 6.7 Hz, 2H), 2.92-3.00 (m, 2H), 3.15-3.32 (m, 2H), 3.41 (t, J= 7.3 Hz, 2H), 3.71 (q, J= 6.0 Hz, 2H), 7.15-7.21 (m, 3H), 7.33-7.37 (m, IH)
e) tert-Butyl [2-(2-chlorophenyl)ethyl] {3-[(2-oxoethyl)thio]propyl}carbamate
Figure imgf000036_0001
The tert-butyl [2-(2-chlorophenyl)ethyl]{3-[(2-hydroxyethyl)thio]propyl}carbamate (leq, 228g, O.όlmol) was dissolved in DMSO (1.5 litres), and treated with triethylamine (lOeq, 850 ml, 6.1mol). The mixture was stirred vigorously, and a solution of sulfur trioxide: pyridine complex (3eq, 291g, 1.83mol) in DMSO (1.5 litres) was added at such a rate that the internal temperature did not exceed 250C (approx 40 minutes). The reaction was poured into a mixture of ice/cone. HCl (~4 litres, 2M), at such a rate as to keep the temperature below 3O0C. This mixture was extracted with tert-butyl methyl ether (2 x 1.5 litres, 1 x 1.2 litres). The organic extracts were combined, washed with water (3 x 1.25 litres), dried over anhydrous magnesium sulphate, filtered and evaporated. The residue was split in to two equal portions, and each passed down a 1 kg pad of silica, eluting with isohexane:ethyl acetate (4:1) to give the desired material (13Og) (57% yield).
1H NMR δ(DMso) 9.40 (t, J= 3.5 Hz, IH), 7.42-7.40 (m, IH), 7.28-7.24 (m, 3H), 3.39-3.33 (m, 2H), 3.21-3.15 (m, 2H), 2.91-2.86 (m, 2H), 2.42-2.33 (m, 2H), 1.73-1.62 (m, 2H), 1.35-1.18 (m, HH).
f) tert-Butyl [2-(2-chlorophenyl)ethyl] {3-[(2-{[(2/?)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3- dihydro-l,3-benzothiazol-7-yl)ethyI]amino}ethyl)thio]propyl}carbamate
Figure imgf000037_0001
TMSCI, NEt3, DCM
Figure imgf000037_0002
2) M, NaBH(OAc)3
34%
Figure imgf000037_0003
To a stirred suspension of 7-[(l /?)-2-amino-l -hydroxy ethyl]-4-hydroxy-l,3-benzothiazol- 2(3H)-one acetate (leq, 92.1g, 0.297 mol) in DCM (1200 ml) was added triethylamine (5.7eq, 237 ml, 1.7 mol) at ambient temperature. Chlorotrimethylsilane (4.4eq, 141g, 164 ml, 1.3 mol) was added in portions over 20 minutes-the first 20 ml caused an exotherm to 4O0C, so an ice/water bath was used to maintain the temperature at 250C. The mixture was stirred for 4 hours at room temperature. Anhydrous magnesium sulphate (98g) was added in a single portion to the reaction mixture, which was stirred for 15 minutes, before a solution of tert-butyl [2-(2-chlorophenyl)ethyl] {3-[(2-oxoethyl)thio]propyl}carbamate (l.leq, 12Og, 0.323 mol) in DCM (800 ml) was added dropwise over 90 minutes. Sodium triacetoxyborohydride (1.2eq, 75g, 0.35 mol) added in one portion, maintaining the temperature at 260C. The mixture was stirred for 16 hours at ambient temperature. Methanol (350 ml) was added in portions, followed by acetic acid (70 ml) and the mixture was stirred for 2 hours at room temperature. The solvent was removed in vacuo, and the acetic acid was removed by means of a toluene (600 ml) azeotrope. The residue was partitioned between water (1000 ml) and ethyl acetate (400 ml), the layers separated and the aqueous layer was further extracted with ethyl acetate (2x 200 ml). The organics were combined and washed with water (400 ml), dried over anhydrous sodium sulfate, filtered and evaporated to give a dark oil (93g). Isohexane (200 ml) was added and the resultant tar was manipulated with a spatula. The isohexane was decanted from the tar, and the process repeated twice more. The residue was divided into 2 batches and purified using silica column chromatography (large Biotage 75) eluting with 5% MeOH in DCM (2.5 column volumes), 10% MeOH in DCM (5 column volumes), then 16% MeOH in DCM (2.5 column volumes) to give the desired material (86.9g) (46% yield).
1H NMR (300 MHz, DMSO) δ 1.27-1.36 (m, 9H), 1.64-1.74 (m, 2H), 2.44-2.49 (m, 2H), 2.75-2.85 (m, 2H), 2.86-3.02 (m, 4H), 3.14-3.23 (m, 2H), 3.32-3.41 (m, 4H), 4.82 (t, J =
6.1 Hz, IH), 6.76 (d, J= 8.4 Hz, IH), 6.90 (d, J= 8.4 Hz, IH), 7.27 (s, 3H), 7.41 (d, J=
7.2 Hz, IH)
HPLC: 92.25% @ 220nm [M+H]+=582 (calc=582.1863) (MultiMode+)
g) 7-[(lR)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l- hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3//)-one Dihydrobromide Polymorph B
Figure imgf000038_0001
Formic acid (54 ml) was mixed with water (6 ml) and was left to age for several hours. tert-Butyl [2-(2-chlorophenyl)ethyl]{3-[(2-{[(2i?)-2-hydroxy-2-(4-hydroxy-2-oxo-2,3- dihydro-l,3-benzothiazol-7-yl)ethyl]amino}ethyl)thio]propyl}carbamate (6.Og, 10.3mmol) was dissolved in the aqueous formic acid and stirred at room temperature for 18 hours. The solvents were removed in vacuo, and the residue was dissolved in a 4:1 mixture of acetonitrile:water (10 ml), filtered, washed with more 4:1 acetonitrile:water (4 ml), then purified using reversed phase HPLC on a 30 x 100 Sunfire column, injecting 2ml (500mg) per run, and eluting with 5-50% acetonitrile in 0.2% aqueous TFA over 8 minutes, collecting 15ml fractions. The appropriate fractions were combined and evaporated to give 5.88g. This material was dissolved in acetonitrile (120 ml) (sometimes a suspension was formed) and acidified with a 30% solution of 48% aq HBr in acetonitrile (25 ml). The resulting suspension was agitated, left for 15 minutes, then the solid was collected by filtration, washed with acetonitrile (x5) and dried to give the desired material (4.45g) (67% yield) as a highly crystalline solid identified by XPRD as Polymorph B.
HPLC: 98.42% @ 220nm [M+H]+=482.1 (calc=482.1339) (MultiMode+)
Elemental analysis: C H N S
Calculated: 41 .04 4 .70 6.53 9.96
Found: 40 .81 4 .72 6.73 10.4
Enantiomeric purity: 97. 58%
Preparation 3
T-IClRJ-Z-dZ-ICS-I^^Z-ChlorophenyOethyllaminoJpropyOthiolethylJ-aiiiino)-!- hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3//)-one hydrochloride - Type A
A 37 wt/wt% solution of hydrochloric acid (175.77 μL) was added to a suspension of 7- [( 1 R)-2-( {2- [(3 - { [2-(2-chlorophenyl)ethyl]amino } propyl)thio]ethyl } -amino)- 1 - hydroxy ethyl]-4-hydroxy-l,3-benzothiazol-2(3H)-one (0.5 g) in methanol (5 mL). The mixture was sonicated then stirred at room temperature for 16 h. The solvent was then removed in vacuo and the residue was treated with ethyl acetate (20 mL) and stirred at room temperature for 1 h. The title compound was isolated by filtration, washed with ethyl acetate (5 mL) and dried in vacuo (0.45 g). 1H NMR (300 MHz, DMSO) δ 7.45 (m, 2H), 7.32 (m, 2H), 6.93 (d, IH), 6.79 (d, IH), 4.98 (m, IH), 3.16 (m, 6H), 3.03 (m, 4H), 2.84 (t, 2H), 2.68 (t, 2H), 1.96 (m, 2H). Enantiomeric purity: 96.7% (R); 3.3% (S).
Preparation 4 7-[(lR)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyl}-amino)-l- hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3//)-one hydrochloride - Type B 20mg of Type A material (Example 2) was placed into a vial, to which was added ethanol (ImI). The mixture was left to stir at room temperature in a capped vial for one week. The resulting suspension was then centrifuged and the solid collected and left to dry overnight in a fume hood.
Preparation 5
7-[(lR)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyl}-amino)-l- hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3//)-one hydrochloride - Type C
20mg of Type A material (Example 2) was placed into a vial, to which was added water (ImI). The mixture was left to stir at room temperature in a capped vial for one week. The resulting suspension was then centrifuged and the solid collected and left to dry overnight in a fume hood.
Preparation 6 7-[(lR)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyl}-amino)-l- hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3//)-one monoxinafoate
l-Hydroxy-2-naphthoic Acid (394.31 mg) was added to a suspension of 7-[(l/?)-2-({2-[(3- {[2-(2-chlorophenyl)ethyl]amino}propyl)thio]ethyl}-amino)-l-hydroxyethyl]-4-hydroxy- l,3-benzothiazol-2(3H)-one (0.5 g) in methanol (5 mL). The mixture was sonicated then stirred at room temperature for 16 h. The solvent was then removed in vacuo and the residue was treated with ethyl acetate (20 mL) and stirred at room temperature for 1 h. The mixture was filtered but no solid material could be isolated so the material collected on the filter and the filtrates were recombined in methanol then evaporated to dryness. The residue was stirred in diethyl ether (30 mL) for 2 h. The title compound was isolated by filtration, washed with diethyl ether (10 mL) and dried in vacuo to leave a non-crystalline product (0.53 g).
1H NMR (300 MHz, DMSO) δ 8.20 (d, IH), 7.74 (m, 2H), 7.48 (m, 2H), 7.40 (m, 2H), 7.33 (m, 2H), 7.04 (d, IH), 6.93 (d, IH), 6.78 (d, IH), 4.95 (m, IH), 3.16 - 3.00 (m, 10H), 2.83 (m, 2H), 2.66 (t, 2H), 1.93 (m, 2H). Enantiomeric purity: 97.3% (R); 2.7% (S). Salt stoichiometry - confirmed as Mono Xinafoate salt by 1H NMR.
Preparation 7
7-[(lR)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyI}-amino)-l- hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3/j)-one monofumarate
Fumaric acid (120.39 mg) was added to a suspension of 7-[(li?)-2-({2-[(3-{[2-(2- chlorophenyl)ethyl]amino}propyl)thio]ethyl} -amino)- 1 -hydroxyethyl]-4-hydroxy- 1 ,3- benzothiazol-2(3H)-one (0.5 g) in methanol (5 mL). The mixture was then stirred at room temperature for 2 h. The solvent was removed in vacuo and the residue was suspended in ethyl acetate (20 mL) and stirred at room temperature for 48 h. The title compound was isolated by filtration, washed with ethyl acetate (5 mL) and dried in vacuo to leave a noncrystalline product (0.59 g).
1H NMR (300 MHz, DMSO) δ 7.43 (m, 2H), 7.31 (m, 2H), 6.91 (d, IH), 6.76 (d, IH), 6.55 (s, 2H), 4.84 (t, IH), 3.07 (s, 4H), 2.96 (m, 6H), 2.74 (t, 2H), 2.62 (t, 2H), 1.90 (quintet, 2H). Enantiomeric purity: 97.2% (R); 2.8% (S). Salt stoichiometry - confirmed as Mono Fumarate salt by 1H NMR.
Preparation 8
7-[(l/f)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyl}-amino)-l- hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3//)-one sulfate
Concentrated sulphuric acid (510.68 μL) was added to a suspension of 7-[(li?)-2-({2-[(3- {[2-(2-chlorophenyl)ethyl]amino}propyl)thio]ethyl}-amino)-l-hydroxyethyl]-4-hydroxy- l,3-benzothiazol-2(3H)-one (0.5 g) in methanol (5 mL). The mixture was sonicated then stirred at room temperature for 2 h. The solvent was removed in vacuo and the residue was suspended in diethyl ether (20 mL) and stirred at room temperature for 1 h. The title compound was isolated by filtration, washed with diethyl ether (5 mL) and dried in vacuo to leave a non-crystalline product. 1H NMR (300 MHz, DMSO) δ 7.45 (m, 2H), 7.32 (m, 2H), 6.93 (d, IH), 6.77 (d, IH), 4.95 (m, IH), 3.50 - 3.00 (m, number of protons could not be determined), 2.83 (m, 2H), 2.65 (m, 2H), 1.92 (m, 2H). Enantiomeric purity: 90.5% (R); 9.5% (S).
Preparation 9
7-[(lΛ)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyl}-amino)-l- hydroxyethyl]-4-hydroxy-l,3-benzothiazol-2(3//)-one citrate
Citric acid (199.27 mg) was added to a suspension of 7-[(l/?)-2-({2-[(3-{[2-(2- chlorophenyl)ethyl]amino}propyl)thio]ethyl}-amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one (0.5 g) in methanol (5 mL). The mixture was sonicated then stirred at room temperature for 2 h. The solvent was removed in vacuo and the residue was suspended in diethyl ether (20 mL) and stirred at room temperature for 1 h. The title compound was isolated by filtration, washed with diethyl ether (5 mL) and dried in vacuo to leave a non-crystalline product.
1H NMR (300 MHz, DMSO) δ 7.44 (m, 2H), 7.33 (m, 2H), 6.93 (d, IH), 6.79 (d, IH), 4.92 (m, IH), 3.32 - 3.01 (m, number of protons could not be determined), 2.79 (m, 2H), 2.67 - 2.49 (m, number of protons could not be determined), 1.91 (m, 2H). Enantiomeric purity: 93.6% (R); 6.4% (S).
Preparation 10
7-[(lR)-2-({2-[(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thiolethyl}-amino)-l- hydroxyethyl]-4-hydroxy-l,3-beiizothiazol-2(3/J)-one phosphate
Phosphoric acid (119.58 mg) was added to a suspension of 7-[(U?)-2-({2-[(3-{[2-(2- chlorophenyl)ethyl]amino } propyl)thio]ethy 1 } -amino)- 1 -hydroxy ethy l]-4-hydroxy- 1,3- benzothiazol-2(3H)-one (0.5 g) in methanol (5 mL). The mixture was then stirred at room temperature for 1 h. The solvent was removed in vacuo and the residue was suspended in diethyl ether (20 mL) and stirred at room temperature for 16 h. The solvent had evaporated so the residue was treated with more diethyl ether (5 mL). The title compound was isolated by filtration, washed with diethyl ether (5 mL) and dried in vacuo to leave a non-crystalline product (0.47g).
1H NMR (300 MHz, DMSO) δ 7.44 (m, 2H), 7.31 (m, 2H), 6.92 (d, IH), 6.76 (d, IH), 4.93 (t, IH), 3.17 - 2.91 (m, 10H), 2.88 - 2.56 (m, 4H), 1.95 (m, 2H). Enantiomeric purity: 93.3% (R); 6.7% (S).
Adrenergic β2 mediated cAMP production
Cell preparation
H292 cells were grown in 225cm2 flasks incubator at 37°C, 5% CO2 in RPMI medium containing, 10% (v/v) FBS (foetal bovine serum) and 2 mM L-glutamine.
Experimental Method Adherent H292 cells were removed from tissue culture flasks by treatment with
Accutase™ cell detachment solution for 15 minutes. Flasks were incubated for 15 minutes in a humidified incubator at 37°C, 5% CO2. Detached cells were re-suspended in RPMI media (containing 10% (v/v) FBS and 2 mM L-glutamine) at 0.05 x 106 cells per mL. 5000 cells in 100 μL were added to each well of a tissue-culture-treated 96-well plate and the cells incubated overnight in a humidified incubator at 37°C, 5% CO2. The culture media was removed and cells were washed twice with 100 μL assay buffer and replaced with 50 μL assay buffer (HBSS solution containing 1OmM HEPES pH7.4 and 5 mM glucose). Cells were rested at room temperature for 20 minutes after which time 25 μL of rolipram (1.2 mM made up in assay buffer containing 2.4% (v/v) dimethylsulphoxide) was added. Cells were incubated with rolipram for 10 minutes after which time Compound A was added and the cells were incubated for 60 minutes at room temperature. The final rolipram concentration in the assay was 300 μM and final vehicle concentration was 1.6% (v/v) dimethylsulphoxide. The reaction was stopped by removing supernatants, washing once with 100 μL assay buffer and replacing with 50 μL lysis buffer. The cell monolayer was frozen at -80°C for 30 minutes (or overnight). AlphaScreen™ cAMP detection
The concentration of cAMP (cyclic adenosine monophosphate) in the cell lysate was determined using AlphaScreen™ methodology. The frozen cell plate was thawed for 20 minutes on a plate shaker then 10 μL of the cell lysate was transferred to a 96- well white 5 plate. 40 μL of mixed AlphaScreen™ detection beads pre-incubated with biotinylated cAMP, was added to each well and the plate incubated at room temperature for 10 hours in the dark. The AlphaScreen™ signal was measured using an EnVision spectrophotometer (Perkin-Elmer Inc.) with the recommended manufacturer's settings. cAMP concentrations were determined by reference to a calibration curve determined in the same experiment
I0 using standard cAMP concentrations. A concentration response curve for Compound A was constructed and data was fitted to a four parameter logistic equation to determine both the PEC50 and Intrinsic Activity. Intrinsic Activity was expressed as a fraction relative to the maximum activity determined for formoterol in each experiment. A result for Compound A is in Table 1. is
Selectivity Assays
Adrenergic αlD o Membrane Preparation
Membranes were prepared from human embryonic kidney 293 (HEK293) cells expressing recombinant human αlo receptor. These were diluted in Assay Buffer (5OmM HEPES, ImM EDTA, 0.1% gelatin, pH 7.4) to provide a final concentration of membranes that gave a clear window between maximum and minimum specific binding. 5
Experimental Method
Assays were performed in U-bottomed 96-well polypropylene plates. 10 μL [3H] -prazosin (0.3 nM final concentration) and 10 μL of Compound A (10x final concentration) were added to each test well. For each assay plate 8 replicates were obtained for [3H]-prazosin0 binding in the presence of 10 μL vehicle (10% (v/v) DMSO in Assay Buffer; defining maximum binding) or lOμL BMY7378 (10 μM final concentration; defining non-specific binding (NSB)). Membranes were then added to achieve a final volume of 100 μL. The plates were incubated for 2 hours at room temperature and then filtered onto PEI coated GF/B filter plates, pre-soaked for 1 hour in Assay Buffer, using a 96-well plate Tomtec cell harvester. Five washes with 250 μL wash buffer (5OmM HEPES, ImM EDTA, pH 7.4) were performed at 4°C to remove unbound radioactivity. The plates were dried then sealed from underneath using Packard plate sealers and MicroScint-0 (50 μL) was added to each well. The plates were sealed (TopSeal A) and filter-bound radioactivity was measured with a scintillation counter (TopCount, Packard BioScience) using a 3-minute counting protocol.
Total specific binding (B0) was determined by subtracting the mean NSB from the mean maximum binding. NSB values were also subtracted from values from all other wells. These data were expressed as percent of Bo. Compound concentration-effect curves (inhibition of [3H]-prazosin binding) were determined using serial dilutions typically in the range 0.1 nM to 10 μM. Data was fitted to a four parameter logistic equation to determine the compound potency, which was expressed as pIC50 (negative log molar concentration inducing 50% inhibition of [3H]-prazosin binding). Result is shown in Table 1 below.
Adrenergic βl
Membrane Preparation
Membranes containing recombinant human adrenergic beta 1 receptors were obtained from Euroscreen. These were diluted in Assay Buffer (5OmM HEPES, ImM EDTA, 12OmM NaCl, 0.1% gelatin, pH 7.4) to provide a final concentration of membranes that gave a clear window between maximum and minimum specific binding.
Experimental Method
Assays were performed in U-bottomed 96-well polypropylene plates. 10 μL [125I]- Iodocyanopindolol (0.036 nM final concentration) and 10 μL of Compound A (10x final concentration) were added to each test well. For each assay plate 8 replicates were obtained for [125I]-Iodocyanopindolol binding in the presence of 10 μL vehicle (10% (v/v) DMSO in Assay Buffer; defining maximum binding) or 10 μL Propranolol (10 μM final concentration; defining non-specific binding (NSB)). Membranes were then added to achieve a final volume of 100 μL. The plates were incubated for 2 hours at room temperature and then filtered onto PEI coated GF/B filter plates, pre-soaked for 1 hour in Assay Buffer, using a 96-well plate Tomtec cell harvester. Five washes with 250 μL wash buffer (5OmM HEPES, ImM EDTA, 12OmM NaCl, pH 7.4) were performed at 4°C to remove unbound radioactivity. The plates were dried then sealed from underneath using Packard plate sealers and MicroScint-0 (50 μL) was added to each well. The plates were sealed (TopSeal A) and filter-bound radioactivity was measured with a scintillation counter (TopCount, Packard BioScience) using a 3-minute counting protocol.
Total specific binding (B0) was determined by subtracting the mean NSB from the mean maximum binding. NSB values were also subtracted from values from all other wells. These data were expressed as percent of B0. Compound concentration-effect curves (inhibition of [125I]-Iodocyanopindolol binding) were determined using serial dilutions typically in the range 0.1 nM to 10 μM. Data was fitted to a four parameter logistic equation to determine the compound potency, which was expressed as pIC5o (negative log molar concentration inducing 50% inhibition of [125I]-Iodocyanopindolol binding). A result is shown in Table 1 below.
Dopamine D2
Membrane Preparation
Membranes containing recombinant human Dopamine Subtype D2s receptors were obtained from Perkin Elmer. These were diluted in Assay Buffer (5OmM HEPES, ImM EDTA, 12OmM NaCl, 0.1% gelatin, pH 7.4) to provide a final concentration of membranes that gave a clear window between maximum and minimum specific binding.
Experimental Method Assays were performed in U-bottomed 96-well polypropylene plates. 30 μL [3H]- spiperone (0.16 nM final concentration) and 30 μL of Compound A (1Ox final concentration) were added to each test well. For each assay plate 8 replicates were obtained for [3H] -spiperone binding in the presence of 30 μL vehicle (10% (v/v) DMSO in Assay Buffer; defining maximum binding) or 30 μL Haloperidol (10 μM final concentration; defining non-specific binding (NSB)). Membranes were then added to achieve a final volume of 300 μL. The plates were incubated for 2 hours at room temperature and then filtered onto PEI coated GF/B filter plates, pre-soaked for 1 hour in Assay Buffer, using a 96-well plate Tomtec cell harvester. Five washes with 250 μL wash buffer (5OmM HEPES, ImM EDTA, 12OmM NaCl, pH 7.4) were performed at 4°C to remove unbound radioactivity. The plates were dried then sealed from underneath using Packard plate sealers and MicroScint-0 (50 μL) was added to each well. The plates were sealed (TopSeal A) and filter-bound radioactivity was measured with a scintillation counter (TopCount, Packard BioScience) using a 3-minute counting protocol.
Total specific binding (B0) was determined by subtracting the mean NSB from the mean maximum binding. NSB values were also subtracted from values from all other wells. These data were expressed as percent of B0. Compound concentration-effect curves (inhibition of [3H]-spiperone binding) were determined using serial dilutions typically in the range 0.1 nM to 10 μM. Data was fitted to a four parameter logistic equation to determine the compound potency, which was expressed as pIC50 (negative log molar concentration inducing 50% inhibition of [3H]-spiperone binding). A result for Compound A is shown in Table 1.
Table 1
Compound β2 pEC50 β2 Int Act αl bind pIC50 βi bind P IC50 D2 bind pIC50
A 9.2 0.8 7.6 6. 9 5.8
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 lippopolvsaccharride (LPS) in the CRL:CD 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 0.1-1 Oμg/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 compound activity on intra-alveolar neutrophil migration after aerosol challenge with lippopolysaccharride (LPS) in the guinea-pig. Male Dunkin-Hartley guinea-pigs (300-60Og) are placed into open fronted guinea-pig holding cones attached at random around a cylindrical aerosol chamber. Guinea-pigs are held in the challenge cones and exposed to an aerosol of vehicle, or LPS at concentrations of 0.1-30μg/ml in 0.9%saline per group Aerosols are generated using 2 jet nebulisers per column with a flow rate of 12 L/m. 10ml of the challenge agent is placed into each nebuliser. Alternatively animals receive an intratracheal dose of 0.1-10μg/kg. This is repeated up to 8 times according to the experimental protocol.
Guinea-pigs 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. Challenged guinea-pigs are killed by anaesthesia overdose (0.5ml Euthetal i.p.) at 4h-24h post challenge. The lungs are then lavaged. After the trachea is exposed and cannulated using a luer fitting cannula (orange =size 8FG), the lungs are lavaged with 3 x 5ml aliquots of Hanks Buffered Salt Solution (HBSS, EDTA -free). The lavaging is performed with gentle massaging of the chest to ensure appropriate agitation of the fluid in the lungs. The washes are harvested into a 15ml conical, polypropylene centrifuge tube, 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 3 Evaluation of compound activity on intra-alveolar neutrophil migration after aerosol challenge with lippopolysaccharride (LPS) in the mouse. Male C57BL/6/J or BALB/C mice (20-35g) are placed in Perspex exposure boxes in groups of up to 20 and exposed to an aerosol of either 0.3 mg/ml LPS or 0.9% w/v saline. The LPS (Sigma, E.Coli, Ref L-3755, Serotype 026:B6, Lot no. 11 lk4078) is made up in 0.9% w/v saline. An aerosol is generated using two jet nebulisers operated at a flow rate of 12 L/min (6L/min for each nebuliser) for 15 min. Alternatively animals receive an intratracheal dose of 0.1-1 Oμg/kg. This may be repeated up to 8 times.
Mice 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.
Mice are killed with an overdose of Euthatal i.p 30 minutes, l-24hr after LPS challenge. When circulation has ceased, the trachea is cannulated (Portex intravenous cannula) and the airways lavaged with 3 x 0.3ml of Isoton II (Beckman Coulter Ref. 8448011 Lot no.25775). For cytospins, lOOμl of the BALF is added to a cytospin funnel and spun, using a ThermoShandon Cytospin model 3 or 4, at 700 rpm for 5 min. Cells on the slide are stained on the Hema-Tek-2000 automatic slide stainer, using Wright-Giemsa stain and differential cell counts carried out to differentiate eosinophils, neutrophils and lymphomononuclear cells (including monocytes, macrophages and lymphocytes). Typically, 200 cells are counted per slide and each cell type expressed as a percentage of the total count. BALF total white cell count is measured using a Sysmex (Sysmex UK, Milton Keynes).
Example 4 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 recorded. 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:
%changeRveh - % change RcnψJ
% bronchoprotection =
VochangeR veh
Where % change Rveh is the mean of the maximum percentage change in airway resistance in the vehicle treated group. Example 5
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 6
Evaluation of Compounds on Antigen induced eosinophilia in ovalbumin sensitised mice.
20-25g male BALB/c mice are sensitized to ovalbumin by i.p administration of 100 μg of grade V ovalbumin (Sigma) adsorbed onto lmg of aluminium hydroxide gel mixture (Fisher Scientific UK) in 0.3 ml saline. Groups of mice are pre-dosed with compound if required, a minimum of two weeks after sensitization. They are then dosed daily for 1-8 days as study protocol specified, with test compound or 0.25 ml vehicle.
Each day of the 1-8 days, 1 hour after dosing, the mice are placed in perspex chambers (20x11x1 lcm, 10 mice max./chamber) and administered an aerosol challenge of 20mg ml'1 ovalbumin for 36 min (8 ml for 18 min followed by another 8 ml for 18 min). Aerosol delivery is achieved using a DeVilbiss jet nebulizer with a flow rate of 61 min"1. 24h after the last dose the mice are killed with euthatal 0.2 ml i.p. and blood samples are taken (in EDTA tubes) for differential cell count analysis, the trachea is cannulated using a pink luer mount Portex cannula cut to lcm and the lungs are lavaged using 3 washes of ImI of Isoton IL. For cytospins, lOOμl of the BALF is added to a cytospin funnel and spun, using a ThermoShandon Cytospin model 3 or 4, at 700 rpm for 5 min. Cells on the slide are stained on the Hema-Tek-2000 automatic slide stainer, using Wright-Giemsa stain and differential cell counts carried out to differentiate eosinophils, neutrophils and lymphomononuclear cells (including monocytes, macrophages and lymphocytes). Typically, 200 cells are counted per slide and each cell type expressed as a percentage of the total count. BALF total white cell count is measured using a Sysmex (Sysmex UK, Milton Keynes).
Example 7
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 8 Evaluation of bronchodilator activity in the guinea pig isolated tracheal ring preparation: onset measurements.
Guinea pigs (300-50Og) were killed by cervical dislocation and the trachea was isolated. The trachea was cut into segments 2-3 cartilage rings in width and suspended in 10ml organ baths in modified Krebs' solution (mM; NaCl, 90; NaHCO3, 45; KCl, 5; MgSO4JH2O, 0.5; Na2HPO4.2H2O, 1 ; CaCl2, 2.25; glucose, 10; pH 7.4 gassed with 5% CO2, 95% O2 at 37°C). The tracheal rings were attached to an isometric force transducer for the measurement of isometric tension. The tissues were washed and a force of Ig was applied to each tissue. The rings were contracted with methacholine (1 μM). Once the contraction had reached a plateau, vehicle (0.01% DMSO in distilled H2O), compound A (0.1 nM), roflumilast ( 1 μM) or a combination of compound A (0.1 nM) and roflumilast (lμM) was added and the tissue left until the response had reached a plateau. Data were collected using the Chart 4 software (ADInstruments, Charlgrove, UK). The time to 90% of the maximum effect of compound A (onset time) was measured for each tissue and expressed in min (mean ± s.e.mean). The onset time for Compound A at O.lnM was 44±9.3 min, in the presence of roflumilast (lμM), the onset time for Compound A (O.lnM) was 11±3.1 min and the onset time for roflumilast was 23±14 min (n = 5; Figure 1).

Claims

C L A I M S
1. A pharmaceutical product comprising, in combination, a first active ingredient which is 7- [( 1 R)-2-( {2-[(3 - { [2-(2-Chloropheny l)ethyl]amino } propy l)thio]ethyl } amino)- 1 - hydroxy ethyl]-4-hydroxy-l,3-benzothiazol-2(3H)-one or a salt thereof, and a second active ingredient selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; an antioxidant; a CCRl antagonist; a chemokine antagonist (not CCRl); a corticosteroid; a CRTh2 antagonist; a DPI antagonist; an Ηistone Deacetylase Inducer; an IKK2 inhibitor; a COX inhibitor; a lipoxygenase inhibitor; a leukotriene receptor antagonist; an MPO inhibitor; a muscarinic antagonist which is Aclidinium bromide, Glycopyrrolate, Oxitropium bromide, Pirenzepine, telenzepine or Tiotropium bromide; a p38 inhibitor; a PDE inhibitor; a PPARγ agonist; a protease inhibitor; a Statin; a thromboxane antagonist; a vasodilator; or, an ENAC blocker (Epithelial Sodium-channel blocker).
2. A kit comprising a preparation of a first active ingredient which is 7-[(li?)-2-({2-[(3- {[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4- hydroxy-l,3-benzothiazol-2(3H)-one or a salt thereof, a preparation of a second active ingredient as defined in claim 1 , and optionally instructions for the simultaneous, sequential or separate administration of the preparations to a patient in need thereof.
5 3. A pharmaceutical composition comprising, in admixture; a first active ingredient which is 7-[(li?)-2-({2-[(3-{[2-(2-
Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4-hydroxy-l,3- benzothiazol-2(3H)-one or a salt thereof; a second active ingredient which is selected from: a non-steroidal Glucocorticoid I0 Receptor (GR Receptor) Agonist; an antioxidant; a CCRl antagonist; a chemokine antagonist (not CCRl); a corticosteroid; a CRTh2 antagonist; a DPI antagonist; an
Ηistone Deacetylase Inducer; an IKK2 inhibitor; a COX inhibitor; a lipoxygenase inhibitor; a leukotriene receptor antagonist; an MPO inhibitor; a muscarinic antagonist as defined in claim 1 ; a p38 inhibitor; a PDE inhibitor; a PPARγ agonist; a protease is inhibitor; a Statin; a thromboxane antagonist; a vasodilator; or, an ENAC blocker
(Epithelial Sodium-channel blocker); and a pharmaceutically acceptable carrier, diluent or adjuvant.
4. A method of treating a respiratory disease, which method comprises simultaneously,0 sequentially or separately administering:
(a) a therapeutically effective dose of a first active ingredient which is 7-[(li?)-2-({2- [(3-{[2-(2-Chlorophenyl)ethyl]amino}propyl)thio]ethyl}amino)-l-hydroxyethyl]-4- hydroxy-l,3-benzothiazol-2(3H)-one or a salt thereof;
(b) a therapeutically effective dose of a second active ingredient as defined in claim 15 to a patient in need thereof.
5. A pharmaceutical product, kit or composition or method according to any one of claims 1 to 4 wherein the first active ingredient is in the form of a salt which is a hydrochloride, hydrobromide, trifluoroacetate, sulphate, phosphate, acetate,0 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-di sulfonate or ethane- 1 -(sulfonic acid)-2-sulfonate), isethionate (2-hydroxyethylsulfonate), 2- 5 mesitylenesulphonate or 2-naphthalenesulphonate.
6. A pharmaceutical product, kit or composition or method according to any one of claims 1 to 4 wherein the first active ingredient is in the form of a salt which is a dihydrobromide. 0
7. A pharmaceutical product, kit or composition or method according to any one of claims 1 to 7 wherein the second active ingredient selected from: a non-steroidal Glucocorticoid Receptor (GR Receptor) Agonist; a CCRl antagonist; s a chemokine antagonist (not CCRl); a corticosteroid; an IKK2 inhibitor; a muscarinic antagonist which is Aclidinium bromide, Glycopyrrolate, Oxitropium bromide, Pirenzepine, telenzepine or Tiotropium bromide; o a p38 inhibitor; or, a PDE inhibitor.
8. A pharmaceutical product, kit or composition or method according to any one of claims 1 to 4 wherein the second active ingredient is a CCRl antagonist which is N-5 (2{(2S)-3[{(3R)-l-[(4-chlorophenyl)methyl]-3-pyrrolidinyl}amino]-2- hydroxypropoxy } -4-fluoropheny l)acetamide, N-(2 { (2S)-3 [ { (3 S)- 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,5 S)- 1 -(4-0 chlorobenzy l)-2,5 -dimethy lpiperidin-4-y ljamino } -2-hydroxy-2-methy lpropy ljoxy } -
4-fluorophenyl)acetic acid, (2- { [(2S)-3- { [(3S,4R)- 1 -(4-chlorobenzyl)-3- methylpiperidin-4-yl]amino}-2-hydroxy-2-methylpropyl]oxy}-4-fluorophenyl)acetic acid, (2-{[(2S)-3-{[(3R,4R)-l-(4-chlorobenzyl)-3-methylpiperidin-4-yl]amino}-2- hydroxy-2-methylpropyl]oxy}-4-fluorophenyl)acetic acid, (2-{[(2S)-3-{[(2R,4S,5S)- 1 -(4-chlorobenzy l)-2,5 -dimethy lpiperidin-4-y l]amino } -2-hydroxy-2- methylpropy l]oxy } -4-fluorophenyl)acetic acid, (2- { [(2S)-3 - { [(2R,4R,5 S)- 1 -(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-chlorobenzy l)-2,5- dimethy lpiperidin-4-y 1] amino } -2-hydroxy-2-methylpropy l]oxy } -4- fluorophenyl)acetic acid, Methyl (2-{[(2S)-3-{[l-(4-chlorobenzyl)piperidin-4- y ljamino } -2-hydroxypropy l]oxy } -4-fluorophenyl)propanoate, N- [2-( { 2S } -3 - [( 1 -[4- chlorobenzyl]-4-piperidinyl)amino]-2-hydroxypropoxy)-4-chlorophenyl acetamide, N- [2-( {2S } -3 - [( 1 - [4-chlorobenzy l]-4-piperidinyl)amino] -2-hydroxy-2- methylpropoxy)-4-hydroxyphenyl] acetamide, N- [2-( {2S } -3- [( 1 - [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- methy lpropoxy)-4-hydroxyphenyl] acetamide, N- [5-chloro- [2-( { 2S } -3 - [( 1 - [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-[l-
{(4-chlorobenzyl)-4-piperidinyl } amino] -2-hydroxypropoxy } -4-hydroxyphenyl)-N' - cyclopropyl-urea, N-(2-{(2S)-3-[l-{(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-piperidiny l)amino] -2-hy droxypropoxy )-N- cyclopropylbenzamide, Methyl 2-({2S}-3-[(l-[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 } -4- hydroxyphenyl)acetamide, N-(2-{[(2S)-3-(5-chloro-l'H-spiro[l,3-benzodioxole-2,4'- piperidin]-r-yl)-2-hydroxypropyl]oxy}-4-hydroxyphenyl)acetamide, 2-{[(2S)-3-(5- chloro-lΗ,3H-spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-4- hydroxy-N-methy lbenzamide, 2- { [(2S)-3-(5-chloro- 1 Η,3H-spiro[ 1 -benzofuran-2,4'- piperidin]-r-yl)-2-hydroxypropyl]oxy}-4-hydroxybenzoic acid, N-(2-{[(2S)-3-(5- chloro- 1 Η,3H-spiro[2-benzofuran-l ,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy }-4- s hydroxyphenyl)acetamide;
2- { [(2S)-3-(5-chloro- 1 Η,3H-spiro[2-benzofuran- 1 ,4'-piperidin]- 1 '-yl)-2- hydroxypropyl]oxy}-4-hydroxy-N-methy lbenzamide, N-(2-{[(2S)-3-(5-fluoro-l'H,3H- spiro[ 1 -benzofuran-2,4'-piperidin] - 1 '-yl)-2-hydroxypropy l]oxy } -A- hydroxyphenyl)acetamide, 2- { [(2S)-3-(5-fluoro- 1 Η,3H-spiro[ 1 -benzofuran-2,4'-
I0 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]- 1 '-yl]-2- hydroxypropyl}oxy)-4-hydroxyphenyl]acetamide, N-(2-{[(2S)-3-(5-chloro-l'H,3H- spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy } -4- hydroxyphenyl)urea, 4-fluoro-2-{[(2S)-3-(5-fluoro-l'H,3H-spiro[l-benzofuran-2,4'- i5 piperidin]-l'-yl)-2-hydroxypropyl]oxy}benzoic acid, N-(2-{[(2S)-3-(5-chloro-l'H,3H- spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-4-fluorophenyl)urea, N-(2-{ [(2S)-2-amino-3-(5-fluoro-l Η,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,
20 (αS)-5 -chloro-α- [ [2-(2-hy droxyethy l)phenoxy ] methyl] - Spiro [benzofuran-2(3 H),4'- piperidine]-l'-ethanol, (αS)-5-chloro-α-[[2-(hydroxymethyl)phenoxy]methyl]- Spiro[benzofuran-2(3H),4'-piperidine]-l'-ethanol, N-(2-{[(2S)-3-(5-chloro-l'H,3H- spiro [ 1 -benzofuran-2 ,4'-piperidin] - 1 '-y l)-2-hydroxypropy 1] oxy } - 5 -chloro-4- hydroxyphenyl)acetamide, 2-Chloro-5-{[(25)-3-(5-chloro-l'H,3H-spiro[l-benzofuran-
25 2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-(4-{acetylamino}phenoxy)acetic acid, 5-
{ [(25)-3-(5-Chloro- 1 'H,3H-spiro[ 1 -benzofuran-2,4'-piperidin]- 1 '-yl)-2- hydroxypropyl]oxy}-(4-{acetylamino}phenoxy)acetic acid, {2-Chloro-5-{[(2iS)-3-(5- chloro- 1 'H,3H-spiro[ 1 -benzofiiran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy } -4- [(methylamino)carbonyl]phenoxy} acetic acid, 2-{2-Chloro-5-{[(2S)-3-(5-chloro-
30 1 'H,3H-spiro[l -benzofuran-2,4'-piperidin]- 1 '-yl)-2-hydroxypropyl]oxy }-4-
[(methylamino)carbonyl]phenoxy}-2-methylpropanoic acid, (2-Chloro-5-{[(25)-3-(5- chloro-rH,3H-spiro[l-benzoflιran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-4- { [(3iS)-3 -hydroxypyrrolidin- 1 -y l]carbony 1 } phenoxy)acetic acid, 5 -Chloro-2- { [(2S)-3 - (5-chloro-rH,3//-spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2-hydroxypropyl]oxy}-4- (cyanomethoxy)benzoic acid, 2-{[(2S)-3-(5-chloro-lΗ,3Η-spiro[l-benzofuran-2,4'- piperidin]-r-yl)-2-hydroxypropyl]oxy}-5-chloro-4-(2,2-difluoroethoxy)benzoic, 5-
5 Chloro-2-{ [(2S)-3-(5-chloro- 1 'H,3H-spiro[l -benzofuran-2,4'-piperidin]- 1 '-yl)-2- hydroxypropyl]oxy } -4-(3 ,3 ,3 -trifluoropropoxy)benzoic acid, N-(2- { 3 - [5 -chloro- lΗ,3Η-spiro[l-benzofuran-2,4'-piperidin]-r-yl]propoxy}phenyl)acetamide, Methyl 3- (2-{[(2S)-3-(5-chloro-l 'H3H-spiro[l-benzofuran-2,4'-piperidin]-r-yl)-2- hydroxypropyl]oxy}-4-fluorophenyl)propanoic acid, N-(2-{[(2S)-3-({spiro[indole-2- i o 4'-piperidin]-3( 1 Η)-one } - 1 '-yl)-2-hydroxypropyl]oxy } -4-hydroxypheny l)acetamide, or
(2-{ [(25)-3-(5-Chloro- 1 'H,3H-spiro[l -benzofuran-2,4'-piperidin]- 1 '-yl)-2- hydroxypropyl]oxy}-4-fluorophenyl)methanesulfbnic 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 is ((2R)-l-[[2-[(aminocarbonyl)amino]-4-chlorophenoxy]acetyl]-4-[(4- fluorophenyl)methyl]-2-methylpiperazine monohydrochloride); or CCX634.
9. A pharmaceutical product, kit or composition or method according to any one of claims 1-7 wherein the second active ingredient is a corticosteroid.
20
10. A pharmaceutical product, kit or composition or method according to any one of claims 1-7 wherein the second active ingredient is a muscarinic antagonist which is Aclidinium bromide, Glycopyrrolate, Oxitropium bromide, Pirenzepine, telenzepine or Tiotropium bromide.
25
11. A pharmaceutical product, kit or composition or method as claimed in claim 10 wherein the second active ingredient is a muscarinic antagonist which is tiotropium bromide.
30 12. A pharmaceutical product, kit or composition or method according to any one of claims 1-7 wherein the second active ingredient is a PDE4 inhibitor.
13. Use of a pharmaceutical product, kit or composition according to claims 1-3 or 5-12 in therapy.
14. Use of a pharmaceutical product, kit or composition according to according to claims 1 -3 or 5- 12 in the manufacture of a medicament for the treatment of a respiratory disease.
15. Use according to claim 14, wherein the respiratory disease is chronic obstructive pulmonary disease, asthma, rhinitis, emphysema or bronchitis.
PCT/GB2008/000667 2007-03-01 2008-02-29 Combinations of beta-2-adrenoceptor agonistic benzothiazolone Ceased WO2008104776A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0703999.3 2007-03-01
GB0703999A GB0703999D0 (en) 2007-03-01 2007-03-01 New combination 667

Publications (1)

Publication Number Publication Date
WO2008104776A1 true WO2008104776A1 (en) 2008-09-04

Family

ID=37965758

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2008/000667 Ceased WO2008104776A1 (en) 2007-03-01 2008-02-29 Combinations of beta-2-adrenoceptor agonistic benzothiazolone

Country Status (2)

Country Link
GB (1) GB0703999D0 (en)
WO (1) WO2008104776A1 (en)

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7700782B2 (en) 2006-12-20 2010-04-20 Astrazeneca Ab Compounds 569
US7709511B2 (en) 2005-08-09 2010-05-04 Astrazeneca Ab Benzothiazolone derivatives
WO2011012897A1 (en) * 2009-07-31 2011-02-03 Astrazeneca Ab New combinations for the treatment of asthma
WO2011061527A1 (en) 2009-11-17 2011-05-26 Astrazeneca Ab Combinations comprising a glucocorticoid receptor modulator for the treatment of respiratory diseases
US7951954B2 (en) 2006-03-14 2011-05-31 Astrazeneca Ab Bezothiazol derivatives as Beta2 adrenoreceptor agonists
US8017602B2 (en) 2008-06-18 2011-09-13 Astrazeneca Ab N-(2-(2-(5-hydroxy-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-8-yl)ethylamino)ethyl)-3-(phenethoxy)propanamide derivatives, processes for their preparation, pharmaceutical compositions containing them and their use in therapy
US8058294B2 (en) 2007-02-08 2011-11-15 Astrazeneca Ab Pharmaceutical salts of N-[2-(diethylamino)ethyl]-N-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-1,3-benzothiazol-7-yl)ethyl]amino}ethyl)-3-[2-(1-napthyl)ethoxy]propanamide
WO2012046050A1 (en) 2010-10-07 2012-04-12 Astrazeneca Ab Novel combinations
MD4369C1 (en) * 2011-03-04 2016-04-30 Sosei R&D Ltd Use of glycopyrrolate for treating tachycardia, unit dose, delivery device, method of treatment and prophylaxis of tachycardia
EP3061821A1 (en) 2009-07-22 2016-08-31 Puretech Ventures Methods and compositions for treatment of disorders ameliorated by muscarinic receptor activation
US10265311B2 (en) 2009-07-22 2019-04-23 PureTech Health LLC Methods and compositions for treatment of disorders ameliorated by muscarinic receptor activation
US10925832B2 (en) 2018-09-28 2021-02-23 Karuna Therapeutics, Inc. Compositions and methods for treatment of disorders ameliorated by muscarinic receptor activation

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007027134A1 (en) * 2005-08-29 2007-03-08 Astrazeneca Ab 7-(2-AMINO-1-HYDROXY-ETHYL)-4-HYDROXYBENZOTHIAZOL-2(3H)-ONE-DERIVATIVES AS β2 ADRENOCEPTOR AGONISTS

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007027134A1 (en) * 2005-08-29 2007-03-08 Astrazeneca Ab 7-(2-AMINO-1-HYDROXY-ETHYL)-4-HYDROXYBENZOTHIAZOL-2(3H)-ONE-DERIVATIVES AS β2 ADRENOCEPTOR AGONISTS

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7709511B2 (en) 2005-08-09 2010-05-04 Astrazeneca Ab Benzothiazolone derivatives
US7951954B2 (en) 2006-03-14 2011-05-31 Astrazeneca Ab Bezothiazol derivatives as Beta2 adrenoreceptor agonists
US7700782B2 (en) 2006-12-20 2010-04-20 Astrazeneca Ab Compounds 569
US8058294B2 (en) 2007-02-08 2011-11-15 Astrazeneca Ab Pharmaceutical salts of N-[2-(diethylamino)ethyl]-N-(2-{[2-(4-hydroxy-2-oxo-2,3-dihydro-1,3-benzothiazol-7-yl)ethyl]amino}ethyl)-3-[2-(1-napthyl)ethoxy]propanamide
US8017602B2 (en) 2008-06-18 2011-09-13 Astrazeneca Ab N-(2-(2-(5-hydroxy-3-oxo-3,4-dihydro-2H-benzo[b][1,4]oxazin-8-yl)ethylamino)ethyl)-3-(phenethoxy)propanamide derivatives, processes for their preparation, pharmaceutical compositions containing them and their use in therapy
US10238643B2 (en) 2009-07-22 2019-03-26 PureTech Health LLC Methods and compositions for treatment of disorders ameliorated by muscarinic receptor activation
US10369143B2 (en) 2009-07-22 2019-08-06 PureTech Health LLC Methods and compositions for treatment of disorders ameliorated by muscarinic receptor activation
US10695339B2 (en) 2009-07-22 2020-06-30 PureTech Health LLC Methods and compositions for treatment of disorders ameliorated by muscarinic receptor activation
EP3646870A1 (en) 2009-07-22 2020-05-06 Puretech Health LLC Methods and compositions for treatment of disorders ameliorated by muscarinic receptor activation
EP3061821A1 (en) 2009-07-22 2016-08-31 Puretech Ventures Methods and compositions for treatment of disorders ameliorated by muscarinic receptor activation
US10369144B2 (en) 2009-07-22 2019-08-06 PureTech Health LLC Methods and compositions for treatment of disorders ameliorated by muscarinic receptor activation
US10265311B2 (en) 2009-07-22 2019-04-23 PureTech Health LLC Methods and compositions for treatment of disorders ameliorated by muscarinic receptor activation
WO2011012897A1 (en) * 2009-07-31 2011-02-03 Astrazeneca Ab New combinations for the treatment of asthma
WO2011061527A1 (en) 2009-11-17 2011-05-26 Astrazeneca Ab Combinations comprising a glucocorticoid receptor modulator for the treatment of respiratory diseases
WO2012046050A1 (en) 2010-10-07 2012-04-12 Astrazeneca Ab Novel combinations
MD4369C1 (en) * 2011-03-04 2016-04-30 Sosei R&D Ltd Use of glycopyrrolate for treating tachycardia, unit dose, delivery device, method of treatment and prophylaxis of tachycardia
US10925832B2 (en) 2018-09-28 2021-02-23 Karuna Therapeutics, Inc. Compositions and methods for treatment of disorders ameliorated by muscarinic receptor activation
US10933020B2 (en) 2018-09-28 2021-03-02 Karuna Therapeutics, Inc. Compositions and methods for treating disorders ameliorated by muscarinic receptor activation
US11452692B2 (en) 2018-09-28 2022-09-27 Karuna Therapeutics, Inc. Compositions and methods for treating disorders ameliorated by muscarinic receptor activation
US11471413B2 (en) 2018-09-28 2022-10-18 Karuna Therapeutics, Inc. Compositions and methods for treating disorders ameliorated by muscarinic receptor activation
US11890378B2 (en) 2018-09-28 2024-02-06 Karuna Therapeutics, Inc. Compositions and methods for treating disorders ameliorated by muscarinic receptor activation

Also Published As

Publication number Publication date
GB0703999D0 (en) 2007-04-11

Similar Documents

Publication Publication Date Title
WO2008104776A1 (en) Combinations of beta-2-adrenoceptor agonistic benzothiazolone
US20100029732A1 (en) Combinations of Beta-2-Adrenoceptor Agonistic Benzothiazolone
WO2008096121A1 (en) Combinations of beta-2-adrenoceptor agonistic benzothiazolone
WO2009037503A2 (en) New combination - 012 for the treatment of respiratory diseases
US20100144606A1 (en) Combination 408
WO2011012897A1 (en) New combinations for the treatment of asthma
WO2012085582A1 (en) Compound
US20150141387A1 (en) Pharmaceutical product comprising a p38 kinase inhibitor and a second active ingredient
JP2009543860A5 (en)
WO2012085583A1 (en) New compound
US10759785B2 (en) Compounds
JP2018536693A (en) 2-Phenyl-3- (piperazinomethyl) imidazo [1,2-a] pyridine derivatives as blockers of TASK-1 and TASK-2 channels for treating sleep-related breathing disorders
JP2018538296A (en) Substituted perhydropyrrolo [3,4-c] pyrrole derivatives and uses thereof
CN108484513A (en) It can be used for treating the chloro-pyrazine carboxamides derivatives of the disease as caused by mucous membrane aquation deficiency
US9969709B2 (en) Guanidinobenzoic acid ester compound
CN108863956A (en) The Epithelial sodium channel blocking compound replaced through aralkyl and aryloxy alkyl
US20110124613A1 (en) Novel Combination of Compounds to be Used in the Treatment of Airway Diseases, Especially Chronic Obstructive Pulmonary Disease (COPD) and Asthma
WO2011073662A1 (en) Combination of a benzoxazinone and a further agent for treating respiratory diseases
WO2012046050A1 (en) Novel combinations
WO2011061527A1 (en) Combinations comprising a glucocorticoid receptor modulator for the treatment of respiratory diseases
WO2010004319A1 (en) 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
HK40071094A (en) Compositions for preventing or treating chronic obstructive pulmonary diseases (copd)

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 08718583

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 08718583

Country of ref document: EP

Kind code of ref document: A1