WO2009060203A1

WO2009060203A1 - 3,4,6,7-tetrahydro-1 h-pyrrolo[3,4-d]pyrimidine-2,5-diones and their therapeutic use

Info

Publication number: WO2009060203A1
Application number: PCT/GB2008/003752
Authority: WO
Inventors: Harry Finch; Nicholas Charles Ray; Christine Edwards
Original assignee: Argenta Discovery Limited
Priority date: 2007-11-07
Filing date: 2008-11-07
Publication date: 2009-05-14
Also published as: GB0721866D0

Abstract

A compound of formula (IA) or (IB): wherein A is aryl or heteroaryl; D is oxygen or sulphur; R1, R2, R3 and R5 are independently each hydrogen, halogen, nitro, cyano, C1-C6-alkyl, C2-C6-alkenyl, C2-C6-alkynyl, hydroxy or C1-C6-alkoxy or C2-C6- alkenyloxy,, wherein C1-C6-alkyl and C1-C6-alkoxy can be further substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy and C1-C4-alkoxy; R4 is hydrogen CrC6-alkyl, formyl, aminocarbonyl, mono- or di-C1-C4- alkylaminocarbonyl, C3-C8-cycloalkylcarbonyl, C1-C6-alkylcarbonyl, C1-C6- alkoxycarbonyl, N-(C1-C4-alkylsulfonyl)-aminocarbonyl, N-(C1-C4-alkylsulfonyl)-N-(C1-C4-alkyl)-aminocarbonyl, heteroaryl, heterocycloalkyl, heteroarylcarbonyl or heterocycloalkylcarbonyl; wherein C1-C6-alkyl, mono- and di-C1-C4- alkylaminocarbonyl, C1-C6-alkylcarbonyl, C1-C2-alkoxycarbonyl, heteroaryl and heterocycloalkyl can be substituted with one to three identical or different radicals selected from the group consisting of aryl, heteroaryl, hydroxyl, C1-C4-alkoxy, hydroxycarbonyl, C1-C6-alkoxycarbonyl, aminocarbonyl, mono and di-C1-C4- alkylaminocarbonyl, amino, mono- and di-C1-C4-alkylamino, C1-C4- alkylcarbonylamino, cyano, N-(mono- and di-Ci-C4-alkylamino-d-C4-alkyl)- aminocarbonyl, N-(C1-C4-alkoxy-C1-C4-alkyl)-aminocarbonyl or halogen; -[Linker]- is a divalent linker radical; and M is a moiety having M3 receptor antagonist activity.

Description

3,4,6,7-TETRAHYDRO-1H-PYRROLO[3,4-D]PYRIMIDINE-2,5-DIONES AND THEIR THERAPEUTIC USE

Field of the Invention

This invention relates to a class of compounds having dual activity as inhibitors of human neutrophil elastase activity and antagonists of M3 muscarinic acetylcholine receptor activation. The compounds have a substituted 3,4,6,7- tetrahydro-1H-pyrrolo[3,4-d]pyrimidine-2,5-dione moiety covalently linked to an M3 receptor antagonist moiety via a linker radical. The invention also relates to the use of such compounds in therapy. Background to the invention

Muscarinic acetylcholine receptors

Anti-cholinergic agents prevent the passage of, or effects resulting from the passage of, impulses through the parasympathetic nerves. This is a consequence of the ability of such compounds to inhibit the action of acetylcholine (Ach) by blocking its binding to the muscarinic cholinergic receptors.

There are five subtypes of muscarinic acetylcholine receptors (mAChRs), termed M1 -M5, and each is the product of a distinct gene and each displays unique pharmacological properties. mAChRs are widely distributed in vertebrate organs, and these receptors can mediate both inhibitory and excitatory actions. For example, in smooth muscle found in the airways, bladder and gastrointestinal tract, M3 mAChRs mediate contractile responses (reviewed by Caulfield, 1993, Pharmac. Then, 58, 319 - 379).

In the lungs, muscarinic receptors M1 , M2 and M3 have been demonstrated to be important and are localized to the trachea, the bronchi, submucosal glands and parasympathetic ganglia (reviewed in Fryer and Jacoby, 1998, Am J Resp Crit Care Med., 158 (5 part 3) S 154 - 160). M3 receptors on airway smooth muscle mediate contraction and therefore bronchoconstriction. Stimulation of M3 receptors localised to submucosal glands results in mucus secretion.

Increased signalling through muscarinic acetylcholine receptors has been noted in a variety of different pathophysiological states including asthma and COPD. In COPD, vagal tone may either be increased (Gross etal. 1989, Chest; 96:984-987) and/or may provoke a higher degree of obstruction for geometric reasons if applied on top of oedematous or mucus-laden airway walls (Gross etal. 1984, Am Rev Respir Dis; 129:856-870). In addition, inflammatory conditions can lead to a loss of inhibitory M2 receptor activity which results in increased levels of acetylcholine release following vagal nerve stimulation (Fryer etal, 1999, Life Sci., 64, (6-7) 449-455). The resultant increased activation of M3 receptors leads to enhanced airway obstruction. Thus the identification of potent muscarinic receptor antagonists would be useful for the therapeutic treatment of those disease states where enhanced M3 receptor activity is implicated. Indeed, contemporary treatment strategies currently support regular use of M3 antagonist bronchodilators as first-line therapy for COPD patients (Pauwels et al. 2001 , Am Rev Respir Crit Care Med; 163:1256-1276).

Incontinence due to bladder hypercontractility has also been demonstrated to be mediated through increased stimulation of M3 mAChRs. Thus M3 mAChR antagonists may be useful as therapeutics in these mAChR-mediated diseases. The drug class of antagonists of M3 muscarinic acetylcholine receptor activation is well represented by known structural types and known individual compounds. Specific classes and compounds are discussed in more detail below. Hereafter, antagonists of M3 muscarinic acetylcholine receptor activation will usually be referred to simply as "M3 antagonists". Human neutrophil elastase Human neutrophil elastase (HNE) is a 32 kDa serine proteinase found in the azurophilic granules of neutrophils. It has a role in the degradation of a wide range of extracellular matrix proteins, including fibronectin, laminin, proteoglycans, Type III and Type IV collagens as well as elastin (Bieth, G. In Regulation of Matrix accumulation, Mecham, R. P. (Eds), Academic Press, NY, USA 1986, 217-306). HNE has long been considered to play an important role in homeostasis through repair and disposal of damaged tissues via degradation of the tissue structural proteins. It is also relevant in the defence against bacterial invasion by means of degradation of the bacterial body. In addition to its effects on matrix tissues, HNE has been implicated in the upregulation of IL-8 gene expression and also induces IL- 8 release from the epithelial cells of the lung. In animal models of Chronic Obstructive Pulmonary Disease induced by tobacco smoke exposure both small molecule inhibitors and protein inhibitors of HNE inhibit the inflammatory response and the development of emphysema (Wright, J. L. et al. Am. J. Respir. Crit. Care Med.2002, 166, 954-960; Churg, A. etal. Am. J. Respir. CrIt. Care Med.2003, 168, 199-207). Thus, HNE may play a role both in matrix destruction and in amplifying inflammatory responses in chronic respiratory diseases where neutrophil influx is a characteristic feature. Indeed, HNE is believed to play a role in several pulmonary diseases, including chronic obstructive pulmonary disease (COPD), cystic fibrosis (CF), acute respiratory distress syndrome (ARDS), pulmonary emphysema, pneumonia and lung fibrosis. It is also implicated in several cardiovascular diseases in which tissue remodelling is involved, for example, in heart failure and the generation of ischaemic tissue injury following acute myocardial infarction. COPD is an umbrella term encompassing three different pathological conditions, all of which contribute to limitation of airflow: chronic bronchitis, emphysema and small-airway disease. Generally all three will exist to varying . extents in patients presenting with COPD, and all three may be due to neutrophil- mediated inflammation, as supported by the increased number of neutrophils observed in bronchoalveolar leakage (BAL) fluids of COPD patients (Thompson, A. B.; Daughton, D.; et al. Am. Rev. Respir. Dis. 1989, 140, 1527-1537). The major pathogenic determinant in COPD has long been considered to be the protease-anti- protease balance (also known as the 'elastase:anti-elastase hypothesis'), in which an imbalance of HNE and endogenous antiproteases such as α1 -antitrypsin (Ci₁-AT), Secretory leukocyte protease inhibitor (SLPI) and pre-elafin leads to the various inflammatory disorders of COPD. Individuals that have a genetic deficiency of the protease inhibitor α 1 -antitrypsin develop emphysema that increases in severity over time (Laurrell, C. B.; Erikkson, S Scand. J. CHn. Invest. 1963 15, 132-140). An excess of HNE is therefore destructive, leading to the breakdown of pulmonary morphology with loss of elasticity and destruction of alveolar attachments of airways in the lung (emphysema) whilst simultaneously increasing microvascular permeability and mucus hypersecretion (chronic bronchitis).

Multimeric ligands consist of multiple binding domains which are tethered together through a suitable scaffold. Hence individual binding domains are linked together into a single molecule, increasing the probability that the multimer will bind sequentially in a step-wise manner with multiple active sites resulting in high-affinity interactions (Handl, H. L. etal. Expert Opin. Ther. Targets2004, 8, 565-586; Han, Y. F. et al., Bioorg. Med. Chem. Letts. 1999, 7, 2569-2575). Also, multiple binding interactions (either sequential or parallel) with relatively high off-rates can combine to yield an overall low off-rate for the multimeric ligand. Thus, a molecule consisting of a suitable linker and ligands may be expected to show advantage over the monomeric ligands alone in terms of potency and/or duration of action. Multimeric compounds are unlikely to be orally bioavailable (as predicted by Lipinski's "Rule of 5") which may be advantageous where an inhaled route of administration to the lungs is targeted, since even after inhaled administration, a large proportion of drug is likely to enter the Gl tract. Thus such compounds may be expected to show reduced systemic exposure after inhalation administration and hence an improved toxicity profile over orally administered therapies.

WO2007/129060 (published after the priority date of this case) relates, inter alia, to homodimeric or heterodimeric compounds of formula M-L-M¹ wherein L is a divalent linker radical and M and M¹ are each independently a radical of formula (A') or (B¹):

wherein A is aryl or heteroaryl;

D is oxygen or sulphur;

R¹, R², R³ and R⁵ are independently each hydrogen, halogen, nitro, cyano,

Ci-Cβ-alkyl, C₂-C₆-alkenyl, C₂-C₆-alkynyl, hydroxy or CrCβ-alkoxy or

C₂-C₆-alkenyloxy,, wherein d-Cβ-alkyl and Ci-C₆-alkoxy can be further substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy and Ci-C₄-alkoxy;

R and R⁴ each independently represent a radical of formula -[X]_m-[Alk¹]_p-[Q]_n- [Alk²]_q-[X¹]_k-Z wherein k, m, n, p and q are independently 0 or 1 ;

AIk¹ and AIk² each independently represent an optionally substituted CrC₆ alkylene, or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-) or amino (-NR^A-) link wherein R^A is hydrogen or CrC₃ alkyl;

Q represents (i) -O-, -S-, -S(=O)-, -S(=O)₂-, -S⁺(R^A)-, -N(R^A)-, -N⁺(R^A)(R^B)-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NR^A -, -NR^AC(=O)-, -S(O₂)NR^A-, -NR^AS(O₂)-, -NR^AC(=O)NR^B-, -NR^AC(=NR^A)NR^B-, -C(=NR^D)NR^E-, -NR^EC(=NR^D)-, wherein R^A, R^B, R^D and R^ε are independently hydrogen, CrC₆ alkyl, or C₃-C₆ cycloalkyl, or R^A and R^Bi or R^D and R^E taken together with the nitrogen to which they are attached form a monocyclic heterocyclic ring of 5 to 7 ring atoms which my contain a further heteroatom selected from N, O and S, or (ii) an optionally substituted divalent monor ., or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members;

X represents -(C=O)-, -S(O₂)-, -C(=O)O-, -(C=O)NR^A-, or -S(O₂)NR^A-, wherein R^A is hydrogen, CrC₆ alkyl, or C₃-C₆ cycloalkyl;

X^I represents -O-, -S-, or -NH; and

Z is hydrogen or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members.

In some embodiments of the compounds (A') and (B') of WO2007/129060, the linker radical L is a divalent straight chain, saturated or unsaturated hydrocarbon radical having from 2 to 12 carbon atoms in the said chain, and wherein one or more carbons is replaced by -N⁺(R^P)(R^Q)-, wherein one of R^p and R^Q is HOC(=O)-(C_rC₆ alkyl)-, and the other is hydrogen, C₁-C₆ alkyl, or C₃-C₆ cycloalkyl, HO-(CrC₆ alkyl)-, HOC(=O)-(C_rC₆ alkyl)- or R^AR^BN-(C_rC₆ alkyl)- wherein R^A and R⁵ are independently hydrogen, C₁-C₆ alkyl, or C₃-C₆ cycloalkyl, or HOC(=O)-(CrC₆ alkyl)-. In such compounds, given the presence of the positively charged nitrogen of the radical -N⁺(R^P)(R^Q)-, the carboxyl group in R^p and R^Q is in the negatively charged carboxylate species form. Such compounds are thus neutral, despite the presence of both a positively charged nitrogen and a negatively charged oxygen. The compounds are thus examples of betaines, which are chemical compounds with both a positively charged cationic functional group such as an ammonium ion or phosphonium ion (an "onium ion") which bears no hydrogen atom, and a negatively charged functional group such as a carboxylate group which may not be adjacent to the cationic site. Betaines are examples of the class of compounds known as zwitterions which are electrically neutral but carry formal positive and negative charges on different atoms which may be adjacent or non-adjacent. Brief description of the invention

This invention relates to compounds comprising an HNE inhibitor radical related to those of formulae (A') and (B') referred to above, covalently linked via a linker radical to an M3 antagonist moiety. The compounds may have a zwitterionic or betaine motif in the linker part. The compounds of the invention are useful in the treatment of diseases or conditions in which HNE activity and excessive M3 receptor activity play a part. Such diseases are often inflammatory diseases, and include inflammatory diseases of the lung. The compounds of the invention are particularly useful in the case of topical pulmonary application by inhalation, v_t

Detailed Description of the Invention

In one embodiment, the invention provides a compound of formula (IA) or (IB):

wherein

A is aryl or heteroaryl; D is oxygen or sulphur;

R¹, R², R³ and R⁵ are independently each hydrogen, halogen, nitro, cyano,

Ci-Ce-alkyl, C₂-C₆-alkenyl, C₂-C₆-alkynyl, hydroxy or CrCe-alkoxy or C₂-C₆- alkenyloxy, wherein CrCe-alkyl and Ci-C₆-alkoxy can be further substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy and C_rC₄-alkoxy; R⁴ is hydrogen or an optional substituent;

-[Linker]- is a divalent linker radical; and

M is moiety having M3 receptor antagonist activity.

Compounds of formula (I) above may be prepared in the form of salts, particularly pharmaceutically acceptable salts, N-oxides, hydrates and solvates thereof. Any claim to a compound herein, or reference to "compounds of the invention", compounds with which the invention is concerned", compounds of formula (I), and the like includes salts, N-oxides, hydrates and solvates of such compounds. Compounds of the invention may be useful in the treatment or prevention of diseases in which excessive HNE and excessive M3 receptor stimulation are implicated, for example chronic obstructive pulmonary disease (COPD)₁ chronic , bronchitis, lung fibrosis, pneumonia, acute respiratory distress syndrome (ARDS), pulmonary emphysema, smoking-induced emphysema and cystic fibrosis. Hence other aspects of the invention are (i) a pharmaceutical composition comprising a compound of the invention and a pharmaceutically acceptable carrier or excipient; and (ii) the use of a compound of the invention for the manufacture of a medicament for the treatment or prevention of a disease or condition in which HNE is implicated. Terminology

As used herein, the term "(C_a-C_b)alkyl" wherein a and b are integers refers to a straight or branched chain alkyl radical having from a to b carbon atoms. Thus when a is 1 and b is 6, for example, the term includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl and n-hexyl. As used herein the term "(C_a-C_b)alkenyl" wherein a and b are integers refers to a straight or branched chain alkenyl moiety having from a to b carbon atoms having at least one double bond of either E or Z stereochemistry where applicable. Thus when a is 2 and b is 6, for example, the term includes, for example, vinyl, allyl, 1 - and 2-butenyl and 2-methyl-2-propenyl. As used herein the term "C_a-C_b alkynyl" wherein a and b are integers refers to straight chain or branched chain hydrocarbon groups having from a to b carbon atoms and having in addition one triple bond. Thus when a is 1 and b is 6, for example, the term includes for example, ethynyl (-C≡CH), 1-propynyl, 1- and 2-butynyl, 2-methyl-2-propynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 2-hexynyl,

3-hexynyl, 4-hexynyl and 5-hexynyl.

As used herein the term "divalent (C_a-C_b)alkylene radical" wherein a and b are integers refers to a saturated hydrocarbon chain having from a to b carbon atoms and two unsatisfied valences.

As used herein the term "divalent (C_a-C_b)alkenylene radical" wherein a and b are integers refers to a divalent hydrocarbon chain having from 2 to 6 carbon atoms, and at least one double bond.

As used herein the unqualified term "carbocyclic" refers to a mono-, bi- or tricyclic radical having up to 16 ring atoms, all of which are carbon, and includes aryl and cycloalkyl.

As used herein the unqualified term "cycloalkyl" refers to a monocyclic saturated carbocyclic radical having from 3-8 carbon atoms and includes, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. As used herein the unqualified term "aryl" refers to a mono-, bi- or tri-cyclic carbocyclic aromatic radical, and includes radicals having two monocyclic carbocyclic aromatic rings which are directly linked by a covalent bond. Illustrative of such radicals are phenyl, biphenyl and napthyl.

As used herein the unqualified term "heteroaryl" refers to a mono-, bi- or tri- cyclic aromatic radical containing one or more heteroatoms selected from S, N and

O, and includes radicals having two such monocyclic rings, or one such monocyclic ring and one monocyclic aryl ring, which are directly linked by a covalent bond.

Illustrative of such radicals are thienyl, benzthienyl, furyl, benzfuryl, pyrrolyl, imidazolyl, benzimidazolyl, thiazolyl, benzthiazolyl, isothiazolyl, benzisothiazolyl, pyrazolyl, oxazolyl, benzoxazolyl, isoxazolyl, benzisoxazolyl, isothiazolyl, triazolyl, benztriazolyl, thiadiazolyl, oxadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, indolyl and indazolyl.

As used herein the unqualified term "heterocyclyl" or "heterocyclic" or

"heterocycloalkyl" includes "heteroaryl" as defined above, and in its non-aromatic meaning relates to a mono-, bi- or tri-cyclic non-aromatic radical containing one or more heteroatoms selected from S, N and O, and to groups consisting of a monocyclic non-aromatic radical containing one or more such heteroatoms which is covalently linked to another such radical or to a monocyclic carbocyclic radical. Illustrative of such radicals are pyrrolyl, furanyl, thienyl, piperidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, pyrazolyl, pyridinyl, pyrrolidinyl, pyrimidinyl, morpholinyl, piperazinyl, indolyl, morpholinyl, benzfuranyl, pyranyl, isoxazolyl, benzimidazolyl, methylenedioxyphenyl, ethylenedioxyphenyl, maleimido and succinimido groups.

Unless otherwise specified in the context in which it occurs, the term "substituted" as applied to any moiety herein means substituted with up to four compatible substituents, each of which independently may be, for example, (CrC₆)alkyl, cycloalkyl, (C₁-C₆JaIkOXy, hydroxy, hydroxy(Ci-C₆)alkyl, mercapto, mercapto(Ci-C₆)alkyl, (CrC₆)alkylthio, phenyl, monocyclic heteroaryl having 5 or 6 ring atoms, halo (including fluoro, bromo and chloro), trifluoromethyl, trifluoromethoxy, nitro, nitrile (-CN), oxo, -COOH, -COOR^A, -COR^A, -SO₂R^A, -CONH₂, -SO₂NH₂, -CONHR^A, -SO₂NHR^A, -CONR^AR^B, -SO₂NR^AR^B, -NH₂, -NHR^A, -NR^AR^B, -OCONH₂, -OCONHR^A, -OCONR^AR^B, -NHCOR^A, -NHCOOR^A, -NR^BCOOR^A, -NHSO₂OR^A, -NR⁸SO₂OH, -NR^BSO₂OR^A,-NHCONH_2> -NR^ACONH₂, -NHCONHR⁸ -NR^ACONHR^B, -NHCONR^AR^B or -NR^ACONR^AR^B wherein R^A and R⁸ are independently a (Ci-C₆)alkyl, (C₃-C₆) cycloalkyl , phenyl or monocyclic heteroaryl having 5 or 6 ring atoms, or R^A and R^B when attached to the same nitrogen atom form a cyclic amino ring, such as piperidinyl, morpholinyl or piperazinyl. An "optional substituent" may be, inter alia, one of the foregoing substituent groups.

As used herein the term "salt" includes base addition, acid addition and quaternary salts. Compounds of the invention which are acidic can form salts, including pharmaceutically acceptable salts, with bases such as alkali metal hydroxides, e.g. sodium and potassium hydroxides; alkaline earth metal hydroxides e.g. calcium, barium and magnesium hydroxides; with organic bases e.g. N-methyl- D-glucamine, choline tris(hydroxymethyl)amino-methane, L-arginine, L-lysine, N- ethyl piperidine, dibenzylamine and the like. Those compounds (I) which are basic can form salts, including pharmaceutically acceptable salts with inorganic acids, e.g. with hydrohalic acids such as hydrochloric or hydrobromic acids, sulphuric acid, nitric acid or phosphoric acid and the like, and with organic acids e.g. with acetic, tartaric, succinic, fumaric, maleic, malic, salicylic, citric, methanesulphonic, p- toluenesulphonic, benzoic, benzenesunfonic, glutamic, lactic, and mandelic acids and the like. Those compounds (I) which have a basic nitrogen can also form quaternary ammonium salts with a pharmaceutically acceptable counter-ion such as chloride, bromide, acaetate, formate, p-toluenesulfonate, succinate, hemi-succinate, naphthalene-bis sulfonate, methanesulfonate, xinafoate, and the like. For a review on salts, see Handbook of Pharmaceutical Salts: Properties. Selection, and Use by Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002).

The term 'solvate' is used herein to describe a molecular complex comprising the compound of the invention and a stoichiometric amount of one or more pharmaceutically acceptable solvent molecules, for example, ethanol. The term 'hydrate' is employed when said solvent is water. Compounds of the invention which contain one or more actual or potential chiral centres, because of the presence of asymmetric carbon atoms, can exist as a number of diastereoisomers with R or S, stereochemistry at each chiral centre. The._.. invention includes all such diastereoisomers and mixtures thereof.

Individual compounds of the invention may exist in several polymorphic forms and may be obtained in different crystal habits. The compounds may also be administered in the form of prodrugs thereof.

So-called 'pro-drugs' of the compounds of formula (I) are also within the scope of the invention. Thus certain derivatives of the compounds which may be active in their own right or may have little or no pharmacological activity themselves can, when administered into or onto the body, be converted into compounds of formula (I) having the desired activity, for example, by hydrolytic cleavage. Such derivatives are referred to as 'prodrugs'. Further information on the use of prodrugs may be found in Pro-drugs as Novel Delivery Systems. Vol. 14, ACS Symposium Series (T. Higuchi and VJ. Stella) and Bioreversible Carriers in Drug Design. Pergamon Press, 1987 (ed. E. B. Roche, American Pharmaceutical Association; CS. Larsen and J. østergaard, Design and application of prodrugs, In Textbook of Drug Design and Discovery, 3^rd Edition, 2002, Taylor and Francis ).

Prodrugs in accordance with the invention can, for example, be produced by replacing appropriate functionalities present in the compounds of formula (I) with certain moieties known to those skilled in the art as 'pro-moieties' as described, for example, in Design of Prodrugs by H. Bundgaard (Elsevier, 1985). Such examples could be a prodrug of a carboxyl group (such as -CO-O-CH₂-O-COtBu as used in the pivampicillin prodrug of ampicillin), an amide (-CO-NH-CH₂-NAIk₂) or an amidine (-C(=N-O-CH₃)-NH₂).

Also included within the scope of the invention are metabolites of compounds of formula (I) that is, compounds formed in vivo upon administration of the drug. Some examples of metabolites include (i) where the contains a methyl group, an hydroxymethyl derivative thereof (-CH₃ -> -CH₂OH); (ii) where the compound contains an alkoxy group, an hydroxy derivative thereof (-OR -> -OH);

(iii) where the compound contains a carboxylic acid ester group, a carboxylic acid derivative thereof (-COOR -> -COOH);

(iv) where the compound contains a tertiary amino group, a secondary amino derivative thereof (-NR¹R² -> -NHR¹ or -NHR²); (v) where the compound contains a secondary amino group, a primary derivative thereof (-NHR¹ -> -NH₂); (vi) where the compound contains a phenyl moiety, a phenol derivative thereof (-Ph -> -PhOH); and (vii) where the compound contains an amide group, a carboxylic acid derivative thereof (-CONH₂ -> COOH). Structural features of compounds of the invention (A) The HNE Inhibitor part

In the HNE inhibitor part in any compatible combination: The atom D may be O or S, but O is currently preferred. The ring A is aryl or heteroaryl and may be any of those rings listed above as examples of aryl or heteroaryl, especially phenyl and monocyclic heteroaryl having 5 or 6 ring atoms. Specific examples include pyridyl, such as 2- and 3-pyridyl, or pyrimidinyl such as pyrimidin-2-yl, but presently it is preferred that A be phenyl.

R¹ and R² may be selected from any of the substituent types for which they are defined in relation to formula (I), including hydrogen, halogen, nitro, cyano, C_rC₃-alkyl, C₂-C₃-alkenyl, C₂-C₃-alkynyl, hydroxyl or C_rC₃-alkoxy or C₂-C₃- alkenyloxy. Specific examples of such substitutuents include hydrogen, fluoro, chloro, bromo, cyano, methyl, methoxy and -C≡CH. For example, -AR¹R² may be 4-cyanophenyl or 4-ethynylphenyl.

R³ and R⁵ too may be selected from any of the substituent types for which they are defined in relation to formula (I), but in one currently preferred type of compound of the invention R⁵ is hydrogen and R³ is 3-trifluoromethyl, 3-chloro or 3- bromo.

R⁴ is hydrogen or an optional substituent. For example, when not hydrogen, R⁴ may be selected from Ci-C₆-alkyl, formyl, aminocarbonyl, mono- or di-Ci-C₄- alkylaminocarbonyl, C₃-C₈-cycloalkylcarbonyl, Ci-C₆-alkylcarbonyl, Ci-C₆- alkoxycarbonyl, N-(CrC₄-alkylsulfonyl)-aminocarbonyl, N-(C_rC₄-alkylsulfonyl)-N-(Ci- C₄-alkyl)-aminocarbonyl, heteroaryl, heterocycloalkyl, heteroarylcarbonyl or heterocycloalkylcarbonyl; wherein Ci-C₆-alkyl, mono- and Ui-C₁-C₄- alkylaminocarbonyl, d-Ce-alkylcarbonyl, d-Cβ-alkoxycarbonyl, heteroaryl and heterocycloalkyl can be substituted with one to three identical or different radicals selected from the group consisting of aryl, heteroaryl, hydroxyl, Ci-C₄-alkoxy,. hydroxycarbonyl, d-C₆-alkoxycarbonyl, aminocarbonyl, mono and di-Ci-C₄- alkylaminocarbonyl, amino, mono- and di-Ci-C₄-alkylamino, C₁-C₄- alkylcarbonylamino, cyano, N-(mono- and di-C₁-C₄-alkylamino-C₁-C₄-alkyl)- aminocarbonyl, N-(Ci -C₄-alkoxy-Ci-C₄-alkyl)-aminocarbonyl and halogen.

In particular subclasses of compounds of the invention, R⁴ is hydrogen, methyl or ethyl. (B) The -[Linker]- radical This -[Linker]- part joins to HNE inhibitor part of the compound to the M3 receptor antagonist part M. In some embodiments, -[Linker]- has the formula -[X]_m-[Alk¹]_p-[X²]_n-[Alk²]_q-[X¹]_k- wherein k, m, n, p and q are independently 0 or 1 ;

X² represents (i) -O-, -S-, -S(=O)-, -S(=O)₂-, -S⁺(R^A)-, -N(R^A)-, -N⁺(R^A)(R^B)-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NR^A -, -NR^AC(=O)-, -S(O₂)NR^A-, -NR^AS(O₂)-, -NR^AC(=O)NR^B-, -NR^AC(=NR^A)NR^B-, -C(=NR^D)NR^E-, -NR^EC(=NR^D)-, wherein R^A, R^B, R^D and R^E are independently hydrogen, C₁-C₆ alkyl, or C₃-C₆ cycloalkyl, or R^A and R^B' or R^D and R^E taken together with the nitrogen to which they are attached form a monocyclic heterocyclic ring of 5 to 7 ring atoms which my contain a further heteroatom selected from N, O and S, or (ii) an optionally substituted divalent mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members;

X^I represents -O-, -S-, or -NH; Thus -[Linker]- may be a divalent straight chain, saturated or unsaturated hydrocarbon radical having from 2 to 12 carbon atoms in the said chain, and wherein one or more carbons may be replaced by a divalent monocyclic or bicyclic carbocyclic or heterocyclic radical having from 3 to 7 ring atoms in the or each ring, or by -O-, -S-, -S(=O)-, -St=O)₂-, -C(=O)-, -N(R^P)-, -N⁺(R^P)(R^Q)-, -C(=O)O-, -OC(=O)-_f -C(=O)NR^A-, -NR^AC(=O)-, -S(O₂)NR^A-, -NR^AS(O₂)-, -NR^AC(=O)NR^B-, -NR^AC(=NR^A)NR^B-, -C(=NR^D)NR^E-, or -NR^EC(=NR^D)-, wherein R^A, R^B, R^D and R^E are independently hydrogen, CrC₆ alkyl, or C₃-C₆ cycloalkyl, and R^p and R^Q are independently hydrogen, CrC₆ alkyl, or C₃-C₆ cycloalkyl, HO-(CrC₆ alkyl)-, R^AR^BN- (C₁-C₆ alkyl)-, or HOCf=O)-(C₁ -C₆ alkyl)-, or R^Aand R^B, or R^D and R^E, or R^p and R^Q taken together with the nitrogens to which they are attached form a monocyclic heterocyclic ring of 5 to 7 ring atoms which may contain a further heteroatom selected from N, O and S.

When one or more one or more -(CH₂)- groups of the linker framework is or are replaced by a divalent monocyclic or bicyclic carbocyclic or heterocyclic radical, the said radical may be selected from, for example, the following:

The linker framework may have, for example, one of the following structures (A), (B)₁(C), (D), (E), (G) and (E): (CH₂)₂.₅-N(CH₃)-(CH₂)₂.₅- (A)

(CH₂)₂.₅-N⁺(CH₃)₂-(CH '₂2)^2-5 (B)

(CH₂2)/2-5 NH-(C=NH)-NH (CH₂)₂.₅ — (C)

(CH₂

(CH₂W __ _{(CH2 (CH2)2 5}.

(D) (E)

(CH₂)₁.₂-CO-NH-(CH₂)₁.₂-N(CH₃)-(CH₂)₁.₂-NH-CO-(CH₂)₁.₂ (G)

(CH^^^CO-NH-fCH^^.-N^CH^-iCH^^.-NH-CO-CCH^_!^— (H)

In one class of examples of this type of -[Linker]-, m may be 0, p, n and q may each be 1 , k may be 0 or 1 , and Q may be -N(R^A) or -N⁺(R^A)(R^B)-, where R^A, R⁸ AIk¹, AIk², and X¹ is as defined in relation to formula (I). X¹ when present may be, for example, -O-. AIk¹ and AIk² when present may be, for example, -(CH₂W-

Other examples of the radical Q when present in -[Linker]- are those divalent radicals which contains an anionic-cationic pair selected from (i) a negatively charged nitrogen and positively charged nitrogen, (ii) a negatively charged oxygen and a positively charged nitrogen or (iii) a negatively charged nitrogen and a positively charged sulfur or (iv) a negatively charged oxygen and a positively charged sulfur. There are many examples of the functional groups comprised in the types (i) to (iv) ion pairs above, and many ways of arranging the pairs structurally. Examples of particular cases of ion pairings and structural types include cases 1 -9 below. Radical Q, Case 1 : In this case, Q represents a divalent radical of formula:

X

-S⁺(A-Q¹ -(B)₅-Z¹)-, -(R^B)N⁺(A-Q¹ -(B)₃-Z¹)- or N⁺ -(A-Q¹-(B)_S-Z¹),

X wherein s is 0 or 1 ; ring L represents a mono- or bicyclic ring or ring system having 3 to 6 ring atoms in the case of a monocyclic ring and up to 10 ring atoms in the case of a bicyclic ring system, A and B each independently represent an optionally substituted divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic radical, or an optionally substituted Ci-C₆ alkylene or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic link wherein R^A is hydrogen CrC₃ alkyl or aryl(CrC₃ alkyl)-; Q¹ is an anionic divalent radical selected from (1) to (11):

R⁸ is hydrogen, C₁-C₆ alkyl, C₃-C₆ cycloalkyl or aryl(C_rC₃ alkyl)-; and Z¹ is hydrogen, trifiuoromethyl, or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members Radical Q. Case 2:

In this case, Q represents a divalent radical of formula:

wherein ring T represents a mono- or bicyclic ring or ring system having 3 to 6 ring atoms in the case of a monocyclic ring and up to 10 ring atoms in the case of a bicyclic ring system, and Q¹ is an anionic divalent radical selected from those of formulae (1 ) to (11 ) as defined in Case 1. Radical Q, Case 3:

In this case, Q represents a divalent radical of formula:

-S⁺(A-Z¹)-, -N⁺(A-Z¹ )(R^B)- or N⁺ H-(A-Z¹)

wherein ring L is as defined in case 1 , A represents an optionally substituted divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic radical, or an optionally substituted CrC₆ alkylene or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic link wherein R^A is hydrogen, C₁-C₃ alkyl or aryl(CrC₃ alkyl)-; R^B is hydrogen, d-C₆ alkyl, aryl(C_rC₃ alkyl)-, or C₃-C₆ cycloalkyl; and Z¹ is hydrogen, trifluoromethyl, or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members, and wherein radical

-A-Z¹ is substituted by an anionic group selected from those of formulae (12) to (20):

or by an anionic group selected from formulae (1) to (11) as defined in case 1 but; wherein one unsatisfied valency is satisfied by a group -A-Z¹.

Radical Q, Case 4: In this case, Q represents a divalent radical-N (E-A-Q¹ -B-Q²-(D)_S-Z¹)- or

-(R^B)C(E-A-Q¹-B-Q²-(D)_S-Z¹)- wherein E is a bond, -C(=O)-, or -Sf=O)₂-; s is 0 or 1 ;

A, B and D independently represent an optionally substituted divalent monocyclic 3- to 6-membered carbocyclic or heterocyclic radical, or an optionally substituted CrC₆ alkylene, or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6-membered carbocyclic or heterocyclic link wherein R^A is hydrogen, Ci-C₃ alkyl or aryl(d-C₃ alkyl)-; Z is hydrogen, trifluoromethyl, or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members; one of Q¹ and Q₂ is an anionic divalent radical selected from those of formulae (1) to (11) as defined in case 1 , while the other of Q¹ and Q² is a divalent cationic radical selected from those of formulae (21) to (27):

wherein ring K is a 4- to 8-membered saturated ring which may contain 1 or 2 additional heteroatoms N, O or S; ring J is a 5- or 6-membered heteroaromatic ring; R' and R" are each hydrogen, or an optional substituent, or R' and R" taken together . represent a divalent C₁-C₆ alkylene or C₂-C₆ akenylene radical in which up to three carbon atoms may be replaced by N O or S atoms. Radical Q, Case 5:

In this case Q represents a divalent radical of formula -N(E-A-Q¹-B-Z¹)- or -(R^B)C(E-A-Q¹-B-Z¹)-, wherein E is a bond, -C(=O)-, or -S(=O)₂-; A and B independently represent an optionally substituted divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic radical, or an optionally substituted CrC₆ alkylene, or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6-membered carbocyclic or heterocyclic link wherein R^A is hydrogen, CrC₃ alkyl or aryl(CrC₃ alkyl)-; Z¹ is hydrogen, trifluoromethyl, or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members; radical -B-Z¹ is substituted by Q²; Q¹ is selected from divalent anionic radicals (1) to (11) as defined in case 1 and Q² is selected from cationic radicals (12) to (20) as defined in case 3, OR Q¹ is selected from divalent cationic radicals (21 ) to (27) as defined in case 4 and Q² is selected from anionic groups of formulae (12) to (20) as defined in case 3 or anionic groups selected from formulae (1) to (11) as defined in case 1 but wherein one unsatisfied valency is satisfied by a group -A-Z¹. Radical Q, Case 6:

In this case, Q represents -(R⁸JC(A-Z¹)- wherein A represents an optionally substituted divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic radical, or an optionally substituted C₁-C₆ alkylene, or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic link wherein R^A is hydrogen or C₁-C₃ alkyl; R^B is hydrogen, C₁-C₆ alkyl, or C₃-C₆ cycloalkyl; Z¹ is hydrogen, trifluoromethyl or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members; and wherein radical -A-Z¹ is substituted by a group selected from:

(28) (29) wherein R' and R" are each hydrogen, Ci-C₃ alkyl or aryl(d-C₃ alkyi)-, or R' and R" taken together with the nitrogen to which they are attached form a 3- to 8-membered heterocyclic ring, and wherein R'" represents hydrogen or one or more optional substituents.

Radical Q, Case 7:

In this case, Q represents a divalent radical of formula

-(Z²-(B¹)_r-Q²-A¹)C(A-Q¹ -(B)₃-Z¹)- wherein r and s are independently 0 or 1 ; A, A¹, B and B¹ each independently represent an optionally substituted divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic radical, or an optionally substituted CrC₆ alkylene, or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6-membered carbocyclic or heterocyclic link wherein R^A is hydrogen, C₁-C₃ alkyl or aryl(CrC₃ alkyl)-; Z¹ and Z² independently represent hydrogen, trifluoromethyl, or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members; one of Q¹ and Q² is an anionic divalent radical selected from those of formulae (1 ) to (11 ) as defined in Case 1 while the other of Q¹ and Q² is a cationic divalent radical selected from those of formulae (21) to (27) as defined in case 4. Radical Q, case 8: In this case, Q represents a divalent radical of formula -(Z²-A¹)C(A-Q¹ -(B)₃- Z¹)- wherein r and s are independently 0 or 1 ; A, A¹, and B each independently represent an optionally substituted divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic radical, or an optionally substituted d-C₆ alkylene, or C₂- C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6-membered carbocyclic or heterocyclic link wherein R^A is hydrogen, C₁-C₃ alkyl or aryl(Ci-C₃ alkyl)-; Z¹ and Z² independently represent hydrogen, trifluoromethyl, or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members; radical Z²-A*- is substituted by Q²; and radical -B-Z¹ is substituted by Q²; Q¹ is selected from divalent anionic radicals (1 ) to (11 ) as defined in case 1 and Q² is selected from cationic groups (12) to (20) as defined in case 3, OR Q¹ is selected from divalent cationic radicals (21 ) to (27) as defined in case 4 and Q² is selected from anionic groups of formulae (12) to (20) as defined in case 3 or groups of formulae (1) to (11) as defined in case 1 but wherein one unsatisfied valency is satisfied by a group -A-Z¹. Radical Q, Case 9: In this case, Q represents a divalent radical selected from:

(30) (3D wherein R' is hydrogen, Ci-C₃ alkyl or aryl(C_rC₃ alkyl)-. Specific examples -[Linker]- radicals include those present in the compounds of the Examples herein. (C) The M3 receptor Antagonist part

In the compounds of the invention, M is moiety having M3 receptor antagonist activity. M3 receptor antagonists (which are anticholinergic agents) constitute a recognised and effective drug class. Many such specific compounds, and many structural classes of such agents are known from the patent and scientific journal literature.

Any compound having M3 antagonist activity may be used as a template from which to form the moiety M of the compounds of the invention. In many cases, the M3 compound will have a "tail" feature resembling the -[Linker]- part of the compounds of the present invention. The point where that "tail" joins the rest of the M3 antagonist compound will often be a convenient point of attachment for the present -[Linker]-. In general, the point of attachment of the moiety M to the -[Linker] radical will be remote from the interface of interaction between the M3 antagonist compound and the receptor. In those rare cases where that interface is not well understood, or when the M3 antagonist compound does not have a "tail" feature as discussed above, it is a routine matter to choose potentially likely points of attachment and to test the resultant compound for M3 antagonist activity in a suitable receptor binding assay and/or functional assay such as those described in the biological section below.

Exemplary known M3 antagonists which can be linked as described above to the -[Linker]- radical to form compounds of the invention include ipratropium (e.g. as the bromide, CAS 22254-24-6, sold under the name Atrovent), oxitropium (e.g. as the bromide, CAS 30286-75-0) and tiotropium (e.g. as the bromide, CAS 136310- 93-5, sold under the name Spiriva). Also of interest are revatropate (e.g. as the hydrobromide, CAS 262586-79-8) and LAS-34273 which is disclosed in WO01/04118 pirenzepine (CAS 28797-61-7), darifenacin (CAS 133099-04-4, or CAS 133099-07-7 for the hydrobromide sold under the name Enablex), oxybutynin (CAS 5633-20-5, sold under the name Ditropan), terodiline (CAS 15793-40-5), tolterodine (CAS 124937-51 -5, or CAS 124937-52-6 for the tartrate, sold under the name Detrol), otilonium (e.g. as the bromide, CAS 26095-59-0, sold under the name Spasmomen), trospium chloride (CAS 10405-02-4) and solifenacin (CAS 242478-37- 1 , or CAS 242478-38-2 for the succinate also known as YM-905 and sold under the name Vesicare). butyiscopolamine bromide, quinuclidinyl benzilate quinuclidinyl-a- hydroxy- diphenylacetate, 1 ,1-dimethyl-4-diphenylacetoxypiperidinium iodide, ipratropium bromide, nitrocaramiphen hydrochloride, pirenzepine dihydrochloride, scopolamine hydrobromide, telenzepine dihydrochloride, tropicamide, hexamethylene-bis-[dimethyl-(3-phthalimidopropyl) ammonium]bromide, atropine sulfate, glycopyrrolate, scopolamine, benztropine mesylate (1S, 3'R)-quinuclidin-3'-yl 1 -phenyl-1 , 2, 3, 4-tetrahydroisoquinoline-2-carboxylate, tripitramine, cyclopentolate hydrochloride, clozapine, (2R)-N-[I -(4-methyl-3-pentenyl)pipehdin-4-yl]- 2- cyclopentyl-2-hydroxy-2-phenylacetamidθ, (+/-)-terodiline, methoctramine tetrahydrochloride, (2R)-2-[(1 R)-3,3-difluorocyclopentyl]-2-hydroxy-2-phe- nylacetamide, 2-diisopropyl aminoethyl 1 -phenylcyclopentane carboxylate hydrochloride, tolterodine tartrate, cyclohexylmethyl-piperidinyltripheny- I- propioamide, 4-cyclohexyl-alpha-[4-[[4-methoxyphenyl]sulfinyl]-phenyl]-1 - -piperazine acetonitrile, tiquizium bromide, oxitropium bromide, pirenzepine dihydrochloride, revatropate, AQ-RA 721 , DAC 5889, hyoscyamine sulfate, oxybutynin, propantheline bromide, flavoxate, dicyclomine, PD 102807, gallamine triethiodide, himbacine, otenzepad, ^• 5,11-dihydro-11 -[2-[2-[(N,N-dipropylaminomethyl)piperidin-1 - yl]ethylamino- ]-carbonyl]6H-pyrido[2,3-B][1 ,4]benzodiazepin-6-one, pfluorohexahydro-sila-difenidol hydrochloride, cyclohexyl-(4-fluorophenyl- )-(3-N- piperidinopropyl)silanol hydrochloride, olanzapine, methscolpamine bromide, π- trihexyphenidyl hydrochloride, darifenacin, homatropine hydrobromide, 2-ethyl-5-(1 - methyl-1 ,2,5,6-tetrahydropyridyl)-2H-tetrazole maleate, 1 ,3-dihydro-1 -{1 -[piperidin-4- yl]piperidin-4-yl}-2H-benzimidizol- -2-ones, 1 ,3-dihydro-1 -{4-amino-1 -cyclohexyl}-2H- benzimidazol-2-ones, benzocycloalkylenylamine derivatives, fesoterodine, (2R)-N-[I - (6-aminopyridin-2-ylmethyl)piperidin-4-yl]-2-[(1 R)-3,3-difluoro- cyclopentyl]-2- hydroxy-2-phenylacetamide, 2-[(1 S,3S)-3-sulfonylaminocyclop- entyl]phenylacetamide derivatives, zamifenacin, (2S, 3'R) 3-quinuclidinyl tropate, 3- quinuclidinyl 3-hydroxymethyl 2-phenyl alkanoates, 3-quinuclidinyl 3-hydroxymethyl 2-thienyl alkanoates, 1 -azabicyclo[2.2.2]octan-3-yl 2-aryl-3-azacyclo-2- hydroxypropionates, 3-quinuclidinol esters, unsymmetrical alpha-disubstituted glycolic acids, benztropine, methscopolamine, KRP-197, and tropicamide.

The following Table gives some representative examples of compounds having M3 receptor antagonist activities, and lists the published patents describing the structural classes to which they belong:

The following Table gives some representative examples of compounds having M3 receptor antagonist activities, and lists the published patents describing the structural classes to which they belong: Organization Related Basic Representative Structure Patent

Aldrich WO 2006081327

(Originator);

sanofi-aventis EP 0364941 (Originator);

Pfizer EP 0424021 ;

(Originator); US 5171744

Boehringer EP 0382687 lngelheim

(Originator);

Scios EP 0486734;

(Originator); US 5001160 sanofi-aventis (Originator);

Astellas WO 1995021820 Pharma (Originator);

Lilly

(Originator); Novo Norαϊsk (Originator);

Dainippon JP 1998175862; Sumitomo WO 1996033169 Pharma (Originator);

Astellas WO 1995006635 Pharma (Originator);

Banyu WO 1996033973 (Originator);

Taisho (Originator);

Banyu W0 1999040070

(Originator);

Merck & Co.

(Originator);

Banyu WO 2001007406 (Originator);

Ranbaxy WO 2004014853 (Originator);

Novartis WO 2005000815 (Originator);

Chiesi WO 2006066924 (Originator); Nikem Research;

Anavex Life FR 2897535; Sciences GR 1004208

(Originator);

Chiesi WO 2006066924 (Originator); Nikem Research;

Dompe US 4556653; (Originator); WO 1993013777

Banyu WO 1998005641 ; (Originator); WO 2001021167 Merck & Co. (Originator);

Almirall WO 2006010452 (Originator);

Pfizer (Originator);

Forest; EP 0296721 ;

Lundbeck US 6297262;

(Originator); WO 1997017074

Salvat WO 2002000652

(Originator);

Pfizer (Originator);

Forest; Almirall (Originator);

Nippon

Shinyaku

(Originator);

Nycomed; Pfizer

(Originator); Almirall;

WO 2007109357

Nippon

Boehringer lngelheim

(Originator);

Boehringer lngelheim

(Originator);

Pfizer;

WO 2003078429;

WO 2003084539; WO 2004047796; WO 2004050093; WO 2004054580; WO 2004058233; WO 2004071383; WO 2004071384; WO 2004071522; WO 2004071527; WO 2004084896; WO 2004084897; WO 2004110404; WO 2005009340; WO 2005011615; WO 2005013994; WO 2005023269; WO 2005042526; WO 2005042527; WO 2005053645; WO 2005053646; WO 2005053647; WO 2005094798; WO 2005102344; WO 2006002840; WO 2006021559; WO 2006076222; WO 2006094924; WO 2006117299; WO 2006117353; WO 2007012626; WO 2007017436; WO 2007017437; WO 2007042467; WO 2007045980; WO 2007075858; WO 2007077164

GlaxoSmithKline; Astellas Pharma (Originator);

Bayer; Procter & Gamble; Ardana Bioscience; Pfizer (Originator); Novartis;

Ono; Kyorin (Originator); LG Life Sciences;

Where structures in the above Table are marked with an asterisk, that asterisk indicates a suitable point of attachment to the -[Linker]- radical to form compounds of the invention. Where the asterisk is adjacent a phenyl ring and not immediately adjacent a particular carbon on that ring, all available carbons in the ring are potential points of attachment but the carbons meta- or para- to the bond linking the phenyl ring to the rest of the molecule are more preferred.

In addition to the patent publications listed in the above Table, the following also describe M3 antagonist compounds which can form the moiety M in compounds of the invention: WO01/04118, WO02/51841 , WO02/53564, WO03/00840119. WO03/87094, WO04/05285, WO02/00652, WO03/53966, EP424021 , US Pat. No. 5171744, US Pat. No. 3714357, WO03/33495; and WO2006/081327,

Other M3 antagonist chemotypes are described in the following patent publications, and they too can form the moiety M in compounds of the invention:

1. WO2007/017670 which relates to M3 antagonist compounds of formula (50);

wherein

A is an oxygen atom or group -N(R¹²)-;

(i) R¹ is CrC₆-alkyl or a hydrogen atom; and R² is a hydrogen atom or a group -R⁵, -Z-Y-R⁵,-Z-NR⁹R¹⁰; -Z-CO-NR⁹R¹⁰; -Z-NR⁹-CO-R⁵; or

-Z-CO₂H; and R³ is a lone pair, or d-Cβ-alkyl in which case the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge; or

(ii) R¹ and R³ together with the nitrogen to which they are attached form a heterocycloalkyl ring, and R² is a hydrogen atom; or a group -R⁵, -Z-Y-

R⁵, -Z-NR⁹R¹⁰, -Z-CO-NR⁹R¹⁰, -Z-NR⁹-CO-R⁵, or -Z-CO₂H, in which cases the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge; or

(iii) R¹ and R² together with the nitrogen to which they are attached form a heterocycloalkyl ring, said ring being substituted by a group -Y-R⁵, -Z-Y-R⁵, -Z-NR⁹R¹⁰; -Z-CO-NR⁹R¹⁰; -Z-NR⁹-CO-R⁵; or -Z-CO₂H and R³ is a lone pair, or d-C_θ-alkyl in which case the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge; R⁴ is a group of formula (a), (b), (c) or (d);

(a) (b) (C) (d)

Z is a C_rCi₆-alkylene, C₂-Ci₆-alkenylene or C₂-Ci₆-alkynylene group;

Y is a bond or oxygen atom;

R⁵ is an d-C₆-alkyl, aryl, aryl-fused-cycloalkyl, aryl-fused-heterocycloalkyl, heteroaryl, aryl(Ci-C₈-alkyl)-, heteroaryl(CrC₈-alkyl)-, cycloalkyl or heterocycloalkyl group;

R⁶ is CrC₆-alkyl or a hydrogen atom;

R and R ,7b are a Ci-C₆-alkyl group or halogen; n and in are independently 0, 1 , 2 or 3; R⁸⁸ and R^8b are independently selected from the group consisting of aryl, aryl- fused-heterocycloalkyl, heteroaryl, d-Cβ-alkyl, cycloalkyl;

R⁸⁰ is -OH, d-Cβ-alkyl, hydroxy-CrC₆-alkyl, nitrile, a group CONR^8d ₂ or a hydrogen atom;

R^M is d-Ce-alkyl or a hydrogen atom; R⁹ and R¹⁰ are independently a hydrogen atom, d-C₆-alkyl, aryl, aryl-fused- heterocycloalkyl, aryl-fused-cycloalkyl, heteroaryl, aryl(d-C₆-alkyl)-, or heteroaryl (d-Cβ-alkyl)- group; or R⁹ and R¹⁰ together with the nitrogen atom to which they are attached form a heterocyclic ring of 4-8 atoms, optionally containing a further nitrogen or oxygen atom; R¹² is d-C₆-alkyl or a hydrogen atom;

Ar¹ is aryl, heteroaryl or cycloalkyl; Ar² are independently aryl, heteroaryl or cycloalkyl; and Q is an oxygen atom, -CH₂-, -CH₂CH₂- or a bond. One preferred class of compounds of the above WO2007/017670 type consists of those having the formula (51 )

wherein ring A is an optionally substituted phenyl ring, or a monocyclic heterocyclic ring of 5 or 6 ring atoms ring, or a phenyl-fused-heterocycloalkyl ring system wherein the heterocycloalkyl ring is a monocyclic heterocyclic ring of 5 or 6 ring atoms; R^8a is phenyl, thienyl, cyclopentyl or cyclohexyl; R^8b is phenyl; thienyl, cyclopentyl or cyclohexyl; s is 1 , 2, 3, 4, 5, 6 or 7 and t is 0, 1 , 2, 3, 4, 5, 6 or 7 provided that s+t is not greater than 10; Y is a bond or -O-, and X^" is a pharmaceutically acceptable anion. The "tail", i.e. (ring A)-(Ch₂)t-Y-(CH₂)s-, of such compounds is clearly analogous to the -[Linker]- radical of compounds of the present invention, and indeed the quaternary nitrogen of such compounds is a suitable point of attachment to the - [Linker]- radical to form compounds of the invention.

2. WO2007/017669 which relates to M3 antagonist compounds of formula (52):

wherein

(i) R¹ is C_rC₆-alkyl or hydrogen; and R² is hydrogen or a group -R⁷, -Z- Y-R⁷, -Z-NR⁹R¹⁰; -Z-CO-NR⁹R¹⁰, -Z-NR⁹-C(O)O-R⁷, or -Z-C(O)-R⁷; and R³ is a lone pair, or CrC₆-alkyl; or

(ii) R¹ and R³ together with the nitrogen to which they are attached form a heterocycloalkyl ring, and R² is a lone pair or a group -R⁷, -Z-Y-R⁷,

-Z-NR⁹R¹⁰, -Z-CO-NR⁹R¹⁰, -Z-NR⁹-C(O)O-R⁷; or ; -Z-C(O)-R⁷; or (iii) R¹ and R² together with the nitrogen to which they are attached form a heterocycloalkyl ring, said ring being substituted by a group -Y-R⁷, -Z-Y-R⁷, -Z-NR⁹R¹⁰; -Z-CO-NR⁹R¹⁰; -Z-NR⁹-C(O)O-R⁷; or ; -Z-C(O)- R⁷; and R³ is a lone pair, or C_rC₆-alkyl;

R⁴ and R⁵ are independently selected from the group consisting of aryl, aryl- fused-heterocycloalkyl, heteroaryl, C_rC₆-alkyl, cycloalkyl; R⁶ is -OH, Ci-C₆-alkyl, CrC₆-alkoxy, hydroxy-CrC₆-alkyl, nitrile, a group

CONR⁸ ₂ or a hydrogen atom;

A is an oxygen or a sulfur atom; X is an alkylene, alkenylene or alkynylene group;

R⁷ is an CrC₆-alkyl, aryl, aryl-fused-cycloalkyl, aryl-fused-heterocycloalkyl, heteroaryl, aryl(d-C₈-alkyl)-, heteroaryl(Ci-C₈-alkyl)-, cycloalkyl or heterocycloalkyl group;

R⁸ is CrC-₆-alkyl or a hydrogen atom;

Z is a CrCi_θ-alkylene, C₂-Ci₆-alkenylene or C₂-Ci₆-alkynylene group; Y is a bond or oxygen atom; R⁹ and R¹⁰ are independently a hydrogen atom, Ci-C₆-alkyl, aryl, aryl-fused- heterocycloalkyl, aryl-fused-cycloalkyl, heteroaryl, aryl(CrC₆-alkyl)-, or heteroaryl (CrC₆-alkyl)- group; or R⁹ and R¹⁰ together with the nitrogen atom to which they are attached form a heterocyclic ring of 4-8 atoms, optionally containing a further nitrogen or oxygen atom. One preferred class of compounds of the above WO2007/017669 type consists of those having the formula (IA)

wherein A is -O- or -S-; m is 1 or 2; ring A is an optionally substituted phenyl ring, or monocyclic heterocyclic ring of 5 or 6 ring atoms, or phenyl-fused-heterocycloalkyl ring system wherein the heterocycloalkyl ring is a monocyclic heterocyclic ring of 5 or 6 ring atoms; R⁴ is phenyl, thienyl, cyclopentyl or cyclohexyl; R⁵ is phenyl; thienyl, cyclopentyl or cyclohexyl; s is 1 , 2, 3, 4, 5, 6 or 7 and t is 0, 1 , 2, 3, 4, 5, 6 or 7 provided that s+t is not greater than 16; Y is a bond or -O-, and X^" is a pharmaceutically acceptable anion. Again, the "tail", i.e. (ring A)-(Ch₂)I-Y-(CH₂)S-, of such compounds is clearly analogous to the -[Linker]- radical of compounds of the present invention, and indeed the quaternary nitrogen of such compounds is a suitable point of attachment to the -[Linker]- radical to form compounds of the invention.

3. WO2007/068929 which relates to M3 antagonist compounds of formula (54);

wherein

R¹ is CrCβ-alkyl or a hydrogen atom;

R² is d-Cβ-alkyl, a hydrogen atom, -Z-Y-R⁵ or -Z-NR⁹R¹⁰; or NR¹R² is heterocycloalkyl;

R³ is a lone pair or CrC₆-alkyl; R⁴ is a group of formula (a) or (b)

(a) (b)

Z is an alkylene, alkenylene or alkynylene group; Y is a bond or oxygen atom;

R⁵ is an alkyl, alkenyl, aryl, heteroaryl, arylalkyl or heteroarylalkyl group; R⁹ and R¹⁰ are independently a hydrogen atom, alkyl, aryl, heteroaryl, arylalkyl, heteroarylalkyl, or alternatively, R⁹ and R¹⁰ may be joined together with the nitrogen atom to which they are attached to form a heterocyclic ring of 4-8 atoms, optionally containing a further nitrogen or oxygen atom. R⁶ is CrCβ-alkyl or a hydrogen atom; R^7a and R^7b are independently d-C₆-alkyl or halogen; n and m are independently 0, 1 , 2 or 3;

R^8a and R^8b are independently selected from aryl, heteroaryl, CrC₆-alkyl and cycloalkyl;

R⁸⁰ is -OH, d-Ce-alkyl, hydroxy-C_rC₆-alkyl or a hydrogen atom;

A is an oxygen atom or group -N(R¹¹)-; and

R¹¹ is CrCβ-alkyl or a hydrogen atom.

The nitrogen atom shown in formula (54) of the compounds of WO 2007/068929 is again a suitable point of attachment to the -[Linker]- radical to form compounds of the invention.

4 WO/2007/123465 and WO06/112078 which relate respectively to M3 antagonist compounds of the following formulae (55) and (56):

wherein

R¹ represents phenyl, benzimidazolyl, benzthiazolyl, or a 5-6 membered heteroaromatic ring, each of which may be optionally substituted by one or more substituents independently selected from halogen, cyano, nitro, S(O)₂R⁶, NR⁷R⁸, S(O)₂NR⁹R¹⁰, C(O)₂R¹³, NR¹⁴S(O)₂R¹⁵, NR¹⁶C(O)R¹⁷, NR¹⁸C(O)₂R¹⁹, NR²⁰C(O)NR²¹R²², OR²³ and Ci-₆ alkyl, which C_1-6alkyl may optionally be substituted by one or more substituents independently selected from halogen, hydroxyl, Ci_-6 alkoxy, NH₂, NH(C_1-6 alkyl) and N(CL₆ alkyl)₂;

R2 represents a C3-5 cycloalkyl ring, which cycloalky ring may be optionally substituted by one or more substituents independently selected from halogen, S(O)₂R²⁴, NR²⁵R²⁶, S(O)₂NR²⁷R²⁸, C(O)NR²⁹R³⁰, NR³¹S(O)₂R³², NR¹⁶C(O)R¹⁷, NR³³C(O)₂R³⁴, and Ci_-6 alkyl, which C_1-6 alkyl may optionally be substituted by one or more substituents independently selected from halogen, hydroxyl, C_1-6 alkoxy, NH₂, NH(CL₆ alkyl) and N(C_1-6 alkyl)₂; R³ represents Ci_-6alkyl;

R⁴ represents hydrogen or d.₆alkyl;

R⁵ represents hydrogen or C_1-6 alkyl; n is 1 or 2; R⁶, R¹³, R¹⁵, R¹⁷, R¹⁹, R²³, R²⁴, R³², R³⁴ and R³⁵ each independently represent hydrogen or C_1-6alkyl, which Ci_-6alkyl may optionally be substituted by one or more substituents independently selected from halogen, hydroxyl, Ci.₆alkoxy, NH₂, NH(Ci- ₆ alkyl) and N(Ci-₆ alkyl)₂; p28 p29

, π , R³⁰, R³¹and R³³ each independently represent hydrogen, C₂-₆ hydroxyalkyl or Ci-β alkyl, which C_1-6 alkyl may optionally be substituted by one or more substituents independently selected from halogen, Ci_-6 alkoxy, NH₂, NH(C_1-6 alkyl) and N(C_1-6 > aikyl)₂; or any of R⁷ and R⁸, R⁹ and R¹⁰, R¹¹ and R¹², R²¹ and R²⁵, R²⁷ and R²⁸, or R²⁹ and R³⁰ together with the nitrogen atom to which they are both attached, may form a

4-8 membered aliphatic heterocyclic ring, which heterocyclic ring may be optionally substituted by one or more substituents independently selected from halogen, hydroxyl, Ci_-6 alkyl, Ci_-6 hydroxyalkyl, and C_t-6 haloalkyl; and X represents a pharmaceutically acceptable anion of a mono or polyvalent acid. In compounds of this type the R⁴ or R⁵ group is again analogous to the -[Linker]- radical of compounds of the present invention, and therefore the quaternary nitrogen of such compounds is a suitable point of attachment to the

-[Linker]- radical to form compounds of the invention.

In addition, the following structures also have M3 antagonist activity, and they too form the moiety M in compounds of the invention: A. A compound of formula (57):

wherein (i) R¹ is d-Cβ-alkyl; and R² is a group -(Z)_p-R⁷, -Z-Y-R⁷, -Z-NR⁹R¹⁰, -Z-CO-NR⁹R¹⁰, or -Z-C(O)-R⁷; and R³ is a lone pair or R³ is C₁-C₆- alkyl, in which case the nitrogen to which they are attached is quaternary and carries a positive charge; PROVIDED THAT R¹, R² and R³ do not all represent CH₃ and that when R³ is a lone pair then R¹ and R² do not both represent CH₃; or

(ii) R¹ and R³ together with the nitrogen to which they are attached form a heterocycloalkyl ring, and R² is a group -(Z)_p-R⁷, -Z-Y-R⁷, -Z-NR⁹R'⁰, -Z-CO-NR⁹R¹⁰, or -Z-C(O)-R⁷, in which case the nitrogen to which they are attached is quaternary and carries a positive charge; or (iii) R¹ and R² together with the nitrogen to which they are attached form a heterocycloalkyl ring, said ring being substituted by a group -R⁷, -Y- R⁷, -Z-Y-R⁷, -Z-NR⁹R¹⁰, -Z-CO-NR⁹R¹⁰, or -Z-C(O)-R⁷; and R³ is a- lone pair or R³ is CrC₆-alkyl, in which case the nitrogen to which they are attached is quaternary and carries a positive charge; p is O oM ;

R⁴ and R⁵ are independently selected from the group consisting of aryl, aryl- fused-heterocycloalkyl, heteroaryl, CrC₆-alkyl, cycloalkyl; or R⁴ and R⁵ are joined together to form a tricyclic ring so that the group

R⁴R⁵R⁶C- represents the group, where R^6a is -OH, CrC₆-alkyl or a hydrogen torn, and Q is an oxygen atom, -CH₂-, -CH₂CH₂- or a bond;

R⁶ is -OH, Ci-C₆-alkyl, CrC₆-alkoxy, hydroxy-CrC₆-alkyl, nitrile, a group CONR¹³ ₂ or a hydrogen atom; one of W, V and A is N or NR¹¹; another of W, V and A is N, O, S or CR⁸; and the last one of W, V and A is N or CR⁸; PROVIDED THAT when A is an oxygen or sulfur atom and W is a nitrogen atom, then V is not a group CR⁸;

X is an Ci-Ci₂-alkylene, C₂-C₁₂-alkenylene or C₂-Ci ₂-alkynylene group; R⁷ is an CrC₆-alkyl, aryl, aryl-fused-cycloalkyl, aryl-fused-heterocycloalkyl, heteroaryl, aryl(Ci-C₈-alkyl)-, heteroaryl(Ci-C₈-alkyl)-, cycloalkyl or heterocycloalkyl group;

Z is a Ci-Ci₆-alkylene, C₂-Ci₆-alkenylene or C₂-Ci₆-alkynylene group; Y is an oxygen atom, a group -S(O)_n, C(O)O, OC(O), N(R¹²JS(O)₂ or

S(O)₂N(R¹²); n is O, 1 or 2;

R⁹ and R¹⁰ are independently a hydrogen atom, d-C₆-alkyl, aryl, aryl-fused- heterocycloalkyl, aryl-fused-cycloalkyl, heteroaryl, aryl(Ci-C₆-alkyl)-, or heteroaryi (CrCe-alkyl)- group; or R⁹ and R¹⁰ together with the nitrogen atom to which they are attached form a heterocyclic ring of 4-8 atoms, optionally containing a further nitrogen or oxygen atom;

R⁸, R¹¹, R¹² and R¹³ are, independently, hydrogen atom or CrC₆-alkyl group; wherein, unless otherwise specified, each occurrence of alkyl, heterocycloalkyl, aryl, aryl-fused-heterocycloalkyl, heteroaryl, cycloalkyl, alkoxy, alkylene, alkenylene, alkynylene or aryl-fused-cycloalkyl may be optionally substituted; and wherein each alkenylene chain may contain 1 , 2 or 3 carbon-carbon double bonds and each alkynylene chain may contain 1 , 2 or 3 carbon-carbon triple bonds. In compounds of this type the nitrogen is again a suitable point of attachment to the -[Linker]- radical to form compounds of the invention. B. A compound of formula (58):

wherein

(i) R¹ is C_rC₆-alkyl or hydrogen; and R² is a group, -Z-Y-W-R⁷; and R³ is a lone pair or CrC₆-alkyl; or

(ii) R¹ and R³ together with the nitrogen to which they are attached form a heterocycloalkyl ring, and R² is a group -Z-Y-W-R⁷; or (iii) R¹ and R² together with the nitrogen to which they are attached form a heterocycloalkyl ring, said ring being substituted by a group -Y-W-R⁷ or -Z-Y-W-R⁷; and R³ is a lone pair or CrC₆-alkyl; R⁴ and R⁵ are independently selected from the group consisting of aryl, aryl- fused-heterocycloalkyl, heteroaryl, C_rC₆-alkyl and cycloalkyl;

R⁶ is OH₁ Ci-C₆-alkyl, d-Ce-alkoxy, hydroxy-C_rC₆-alkyl, nitrile, a group CONR⁹R¹⁰ or a hydrogen atom;

A is an oxygen or a sulfur atom;

X is a d-Cβ-alkylene, C₂-C₈-alkenylene or C₂-C₈-alkynylene group; W is a direct bond or a CrC₈-alkylene, C₂-C₈-alkenylene or C₂-C₈-alkynylene group;

R⁷ is an Ci-C₆-alkyl, aryl, aryl-fused-cycloalkyl, aryl-fused-heterocycloalkyl, heteroaryl, cycloalkyl or heterocycloalkyl group;

R⁸, R⁹, R¹⁰ and R¹¹ are each independently selected from Ci-C₆-alkyl or a hydrogen atom;

Z is a CrCiβ-alkylene, C₂-Ci₆-alkenylene or C₂-Ci₆-alkynylene group; and Y is -S-, -SO-, -SO₂-, -CO₂-, -OC(=O)-, -N(R¹¹)SO₂- or -SO₂N(R¹¹)-. In compounds of this type the nitrogen is again a suitable point of attachment to the -[Linker]- radical to form compounds of the invention. C. A compound of formula (59):

wherein

R² is a group H, -(Z)_p-R⁷, -Z-Y-R⁷ or -Y-R⁷; p is O or 1 ;

R⁴ and R⁵ are independently selected from the group consisting of aryl, aryl- fused-heterocycloalkyl, heteroaryl, CrCβ-alkyl and cycloalkyl;

R⁶ is -OH, Ci-C₆-alkyl, CrC₆-alkoxy, hydroxy-CrC₆-alkyl, nitrile, a group CON R¹R⁹ or a hydrogen atom; one of W, V and A is N or NR¹¹; another of W, V and A is N, O, S or CR⁸; and the last one of W, V and A is N or CR⁸;

X is an CrC₄-alkylene, C₂-C₄-alkenylene or C₂-C₄-alkynylene group;

R⁷ is an d-Ce-alkyl, aryl, aryl-fused-cycloalkyl, aryl-fused-heterocycloalkyl, heteroaryl, aryl(Ci-C₈-alkyl)-, heteroaryl(d-C₈-alkyl)-, or cycloalkyl group; t, u and v are independently selected from 1 , 2 or 3, with the proviso that t, u and v cannot all simultaneously be 1 ;

Z is a C₁-C₄-alkylene, C₂-C₄-alkenylene or C₂-C₄-alkynylene group;

Y is an oxygen atom or a group -S(O)_n; n is 0, 1 or 2;

R¹, R⁸, R⁹ and R¹¹ are, independently, a hydrogen atom or C_rC₆-alkyl group; and

D^" is a pharmaceutically acceptable counter-ion.

In compounds of this type the available carbon atoms of the bicyclic ring containing the quaternised nitrogen are suitable points of attachment to the -[Linker]- radical to form compounds of the invention.

Specific examples of heterodimeric HNE inhibitor - M receptor antagonist compounds of the invention include those of the Examples herein.

As mentioned above, and as is commonly expected in medicinal chemistry, the compounds of the invention may be administered in appropriate cases as prodrugs. One class of prodrugs relevant to the present compounds is the class of esters of compounds (I) (including dimers thereof as discussed herein) which have a carboxylate, sulfonate or phosphonate group in the radical Q in the -[Linker]- part. Ester prodrugs are well known, and include Ci-C₆ alkyl esters. Utilities

The therapeutic utility of the compounds of the invention is pertinent to any disease that is known to be at least partially mediated by the actions of human neutrophil elastase and M3 receptor signaling activities. For example, the present compounds may be beneficial in the treatment of chronic obstructive pulmonary disease (COPD), cystic fibrosis (CF), acute respiratory distress syndrome (ARDS), pulmonary emphysema, pneumonia and lung fibrosis.

The present invention is also concerned with pharmaceutical formulations comprising, as an active ingredient, a compound of the invention. Other compounds may be combined with compounds of this invention for the prevention and treatment of inflammatory diseases of the lung. Thus the present invention is also concerned with pharmaceutical compositions for preventing and treating inflammatory diseases of the lung comprising a therapeutically effective amount of a compound of the invention and one or more other therapeutic agents.

Suitable therapeutic agents for a combination therapy with compounds of the invention include: (1) a corticosteroid, for example fluticasone or budesonide; (2) a β2-adrenoreceptor agonist, for example salmeterol or formeterol; (3) a leukotriene modulator, for example montelukast or pranlukast; (4) anticholinergic agents, for example selective muscarinic-3 (M3) receptor antagonists such as tiotropium bromide; (5) phosphodiesterase-IV (PDE-IV) inhibitors, for example roflumilast or cilomilast; (6) an antitussive agent, such as codeine or dextramorphan; and (7) a . non-steroidal anti-inflammatory agent (NSAID), for example ibuprofen or ketoprofen.

The weight ratio of the first and second active ingredients may be varied and will depend upon the effective dose of each ingredient. Generally, an effective dose of each will be used.

The magnitude of prophylactic or therapeutic dose of a compound of the invention will, of course, vary with the nature of the severity of the condition to be treated and with the particular compound and its route of administration, and will generally be detrmined by clinical trial as required in the pharmaceutical art. It will also vary according to the age, weight and response of the individual patient. In general, the daily dose range will lie within the range of from about 0.001 mg to about 100 mg per kg body weight of a mammal, preferably 0.01 mg to about 50 mg per kg, and most preferably 0.1 to 10 mg per kg, in single or divided doses. On the other hand, it may be necessary to use dosages outside these limits in some cases.

Another aspect of the present invention provides pharmaceutical compositions which comprise a compound of the invention and a pharmaceutically acceptable carrier. The term "composition", as in pharmaceutical composition, is intended to encompass a product comprising the active ingredient(s), and the inert ingredient(s) (pharmaceutically acceptable excipients) that make up the carrier, as well as any product which results, directly or indirectly, from combination, complexation or aggregation of any two or more of the ingredients, or from dissociation of one or more of the ingredients, or from other types of reactions or interactions of one or more of the ingredients. Accordingly, the pharmaceutical compositions of the present invention encompass any composition made by admixing a compound of the invention, additional active ingredient(s), and pharmaceutically acceptable excipients. The pharmaceutical compositions of the present invention comprise a compound of the invention as an active ingredient or a pharmaceutically acceptable salt thereof, and may also contain a pharmaceutically acceptable carrier and optionally other therapeutic ingredients. The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids including inorganic bases or acids and organic bases or acids.

Any suitable route of administration may be employed for providing a mammal, especially a human, with an effective dosage of a compound of the present . invention. In therapeutic use, the active compound may be administered by any convenient, suitable or effective route. Suitable routes of administration are known to those skilled in the art, and include oral, intravenous, rectal, parenteral, topical, ocular, nasal, buccal and pulmonary. Delivery by inhalation is preferred.

Compositions suitable for administration by inhalation are known, and may include carriers and/or diluents that are known for use in such compositions. The composition may contain 0.01 -99% by weight of active compound. Preferably, a unit dose comprises the active compound in an amount of 1 μg to 10 mg.

The most suitable dosage level may be determined by any suitable method known to one skilled in the art. It will be understood, however, that the specific amount for any particular patient will depend upon a variety of factors, including the activity of the specific compound that is used, the age, body weight, diet, general health and sex of the patient, time of administration, the route of administration, the rate of excretion, the use of any other drugs, and the severity of the disease undergoing treatment.

For delivery by inhalation, the active compound is preferably in the form of microparticles. They may be prepared by a variety of techniques, including spray- drying, freeze-drying and micronisation.

By way of example, a composition of the invention may be prepared as a suspension for delivery from a nebuliser or as an aerosol in a liquid propellant, for example for use in a pressurised metered dose inhaler (PMDI). Propellants suitable for use in a PMDI are known to the skilled person, and include CFC-12, HFA-134a, HFA-227, HCFC-22 (CCI₂F₂) and HFA-152 (CH₄F₂ and isobutane)

In a preferred embodiment of the invention, a composition of the invention is in dry powder form, for delivery using a dry powder inhaler (DPI). Many types of DPI are known.

Microparticles for delivery by administration may be formulated with excipients that aid delivery and release. For example, in a dry powder formulation, microparticles may be formulated with large carrier particles that aid flow from the DPI into the lung. Suitable carrier particles are known, and include lactose particles; they may have a mass median aerodynamic diameter of greater than 90 μm. In the case of an aerosol-based formulation, a preferred composition is: Compound of the invention 24 mg / canister

Lecithin, NF Liq. Cone. 1.2 mg / canister

Trichlorofluoromethane, NF 4.025 g / canister Dichlorodifluoromethane, NF 12.15 g / canister.

Compounds of the invention may be used in combination with other drugs that are used in the treatment/prevention/suppression or amelioration of the diseases or conditions for which present compounds are useful. Such other drugs may be administered, by a route and in an amount commonly used therefore, contemporaneously or sequentially with a compound of the invention. When a compound of the invention is used contemporaneously with one or more other drugs, a pharmaceutical composition containing such other drugs in addition to the compound of the invention is preferred. Accordingly, the pharmaceutical compositions of the present invention include those that also contain one or more other active ingredients, in addition to a compound of the invention.

The agents of the invention may be administered in inhaled form. Aerosol generation can be carried out using, for example, pressure-driven jet atomizers or ultrasonic atomizers, preferably using propellant-driven metered aerosols or propellant-free administration of micronized active compounds from, for example, inhalation capsules or other "dry powder" delivery systems.

The active compounds may be dosed as described depending on the inhaler system used. In addition to the active compounds, the administration forms may additionally contain excipients, such as, for example, propellants (e.g. Frigen in the case of metered aerosols), surface-active substances, emulsifiers, stabilizers, preservatives, flavorings, fillers (e.g. lactose in the case of powder inhalers) or, if appropriate, further active compounds.

For the purposes of inhalation, a large number of systems are available with which aerosols of optimum particle size can be generated and administered, using an inhalation technique which is appropriate for the patient. In addition to the use of adaptors (spacers, expanders) and pear-shaped containers (e.g. Nebulator®, Volumatic®), and automatic devices emitting a puffer spray (Autohaler®), for metered aerosols, in particular in the case of powder inhalers, a number of technical solutions are available (e.g. Diskhaler®, Rotadisk®, Turbohaler® or the inhalers for example as described EP-A-0505321). Methods of Synthesis

The compounds of the present invention can be prepared according to the procedures of the following general schemes, using appropriate materials, and are further exemplified by the following specific examples. Moreover, by using the procedures described with the disclosure contained herein, one of ordinary skill in the art can readily prepare additional compounds of the present invention claimed herein. The compounds illustrated in the examples are not, however, to be construed as forming the only genus that is considered as the invention. The examples further illustrate details for the preparation of the compounds of the present invention. Those skilled in the art will readily understand that known variations of the conditions and processes of the following preparative procedures can be used to prepare these compounds.

The compounds of the invention may be isolated in the form of their pharmaceutically acceptable salts, such as those described previously herein above. The free acid or base form corresponding to isolated salts can be generated by treatment with a suitable base or acid such as sodium hydroxide, potassium carbonate, acetic acid and hydrochloric acid and extraction of the liberated free acid or base into an organic solvent followed by evaporation. The free form isolated in this manner can be further converted into another pharmaceutically acceptable salt by dissolution in an organic solvent followed by addition of the appropriate acid or base and subsequent evaporation, precipitation, or crystallisation.

It may be necessary to protect reactive functional groups (e.g. hydroxyl, amino, thio or carboxy) in intermediates used in the preparation of compounds of the invention to avoid their unwanted participation in a reaction leading to the formation of the compounds. Conventional protecting groups, for example those described by T. W. Greene and P. G. M. Wuts in "Protective groups in organic chemistry" John Wiley and Sons, 1999, may be used.

Compounds of the invention may be prepared according to the routes illustrated in Schemes 1 and 2.

Scheme 1

N

ι) Bromination ι) Bromination ι) Bromination ii) R¹NH₂, NaHCO₃ II) R¹NH₂, NaHCO₃ II) R¹NH₂, NaHCO₃ CH₃CN CH₃CN CH₃CN

M = M₃ antagonist

Lg = leaving group e g. Br

G = spacer eg (CH_j)_n

W, X₁ Y and Z are reactive groups e g CO₂H, NH₂, halogen etc Scheme 2

The following Examples illustrate the invention.

General Experimental Details:

All solvents and commercial reagents were used as received. Where products were purified using an Isolute® SPE Si Il cartridge, 'Isolute SPE Si cartridge' refers to a pre-packed polypropylene column containing unbonded activated silica with irregular particles with average size of 50 μm and nominal 6OA porosity. Where an Isolute® SCX-2 cartridge was used, 'Isolute® SCX-2 cartridge' refers to a pre-packed polypropylene column containing a non end-capped propylsulphonic acid functionalised silica strong cation exchange sorbent. 'Isolute®

Al-N cartridge' refers to a pre-packed polypropylene column containing neutral alumina with average particle size 50-200 μm and 120 A pore diameter.

'CombiFlash® companion' refers to an automated flash silica chromatography system which uses pre-packed polypropylene (RediSep®) columns containing silica with average particle size 35-70 μm (230-400 mesh).

Preparative HPLC conditions:

HPLC system 1 :

C18-reverse-phase end-capped column (250 x 21.2 mm Gemini column with

5 μm particle size), eluting with a gradient of A: water + 0.1% formic acid; B: acetonitrile + 0.1% formic acid with a flow rate typically 17 ml/min and gradient of

1 %/min increasing in B. UV detection at 254 nm. Compounds were obtained as the formate salt where stated.

HPLC system 2:

C18-reverse-phase end-capped column (250 x 21.2 mm Gemini column with 5 μm particle size), eluting with a gradient of A: water; B: acetonitrile with a flow rate typically 18 ml/min and gradient of 5%/min increasing in B. UV detection at 254 nm.

LC-MS method 1

Waters Platform LC with a C18-reverse-phase column (30 x 4.6 mm

Phenomenex Luna 3 μm particle size), elution with A: water + 0.1% formic acid; B: acetonitrile + 0.1 % formic acid. Gradient:

Gradient - Time flow ml/min %A %B

0.00 2.0 95 5

0.50 2.0 95 5 4.50 2.0 5 95

5.50 2.0 5 95

6.00 2.0 95 5

Detection - MS, ELS, UV (100 μl split to MS with in-line UV detector) MS ionisation method - Electrospray (positive and negative ion) LC-MS method 2

Waters Micromass ZMD with a C18-reverse-phase column (30 x 4.6 mm Phenomenex Luna 3 μm particle size), elution with A: water + 0.1% formic acid; B: acetonitrile + 0.1% formic acid. Gradient:

Gradient - Time flow ml/min %A %B

0.00 2.0 95 5

0.50 2.0 95 5

4.50 2.0 5 95

5.50 2.0 5 95

6.00 2.0 95 5

Detection - MS, ELS, UV (100 μl split to MS with in-line UV detector)

MS ionisation method - Electrospray (positive and negative ion)

LC-MS method 3

Waters Micromass ZQ2000 with a C18-reverse-phase column (100 x 3.0 mm Higgins Clipeus with 5 μm particle size), elution with A: water + 0.1 % formic acid; B: acetonitrile + 0.1% formic acid. Gradient:

Gradient - Time flow ml/mm %A %B

0.00 1.0 95 5

1.00 1.0 95 5 15.00 1.0 5 95

20.00 1.0 5 95

22.00 1.0 95 5

25.00 1.0 95 5

Detection - MS, ELS, UV (100 μl split to MS with in-line UV detector) MS ionisation method - Electrospray (positive ion)

Abbreviations used in the experimental section:

DCM = dichloromethane

HPLC = high performance liquid chromatography IMS = industrial methylated spirits RT = room temperature Rt = retention time THF = tetrahydrofuran Intermediate 1

A solution of anti-(1 S, 2ff)-7-(benzylmethylamino)bicyclo[2.2.1]heptan-2-ol (1.63 g, 7.05 mmol) and di-tert-butyl dicarbonate (1.69 g, 7.75 mmol) in EtOH was de-gassed with Argon. 20% Pd(OH)₂ on carbon (230 mg, 0.71 mmol) was added and the reaction was degassed again with Argon. Hydrogen gas was bubbled through the solution, and the reaction mixture was allowed to stir at RT under a hydrogen balloon for 18 h. The reaction mixture was filtered through Celite and concentrated in vacuo to give Intermediate 1 as an amber oil. LC-MS (Method 1): Rt 2.89 min, m/z 242 [M+H]⁺ Intermediate 2

A solution of Intermediate 1 (3.0 g, 12.43 mmol) in toluene (75 ml) in a round bottom flask was fitted with a short-path distillation apparatus. Methyl (R)- cyclohexylhydroxyphenylacetate (J. Med. Chem. 1977, 20(12), 1612) (3.40 g, 13.67 mmol) and sodium methoxide (671 mg, 12.43 mmol) were added and the mixture was gradually heated under a slight vacuum until distillation of MeOH (b.p. 7O⁰C, external temp. 120°C) was observed. After this fraction distilled, the mixture was heated further until toluene began to distil (external temp. 140°C). After 3 h the mixture was poured into sat. aqueous ammonium chloride and extracted with EtOAc (2 x 100 ml). The combined organics were dried over (Na₂SO₄), filtered and evaporated. Purification using a CombiFlash® companion eluting with a gradient of 0-35% Et₂O-cyclohexane gave the intermediate Boc-protected compound. This was dissolved in DCM (59 ml) and treated with TFA (6 ml). The solution was stirred at RT for 2 h and then the reaction mixture was added slowly to sat. aqueous sodium hydrogen carbonate and extracted with EtOAc. The combined organic extracts were washed with brine, dried (Na₂SO₄), filtered, and concentrated in vacuo to afford

Intermediate 2 in high purity.

Yield: 2.07 g (43%)

LC-MS (Method 2): Rt 2.59 min, m/z 358 [M+H]⁺, 399 [M+H.MeCNf

Intermediate 3

A mixture of 2-(9-bromononyl)isoindole-1 ,3-dione (J. Org. Chem., 1957, 22, 68-70) (102 mg, 0.28 mmol), (5-methylaminomethyloxazol-2-yl)diphenylmethanol

(WO2007/017669) (83 mg, 0.28 mmol) and caesium carbonate (184 mg, 0.56 mmol) in DMF (1 ml) containing water (20 μl_) was stirred under nitrogen at RT for 20 h.

The solvent was removed in vacuo, water (25 ml) was added to the residue and the product was extracted with DCM (3 x 25 ml). These extracts were washed with water (2 x 50 ml) and brine (50 ml) before the organic phase was isolated, dried

(MgSO₄), filtered, and concentrated in vacuo. Purification using an Isolute® Si Il cartridge eluting with 50% EtOAc in cyclohexane, EtOAc then a MeOH/EtOAc gradient afforded Intermediate 3 as a pale yellow oil.

Yield: 80 mg (50%) LC-MS (Method 1 ): Rt 2.87 min, m/z 566 [M+H]⁺

Intermediate 4

Hydrazine hydrate (20 μL, 0.32 mmol) was added to a solution of Intermediate 3 (80 mg, 0.14 mmol) in IMS (1.5 ml) and the stirred mixture was heated at 80°C for 1 h. The solution was allowed to cool, dilute aqueous sodium hydroxide was added and the product was extracted with EtOAc (3 x 20 ml). The extracts were washed with water (2 x 15 ml) and brine (20 ml) before the organic phase was isolated, dried (MgSO₄), filtered and concentrated in vacuo. The crude product was dissolved in MeOH then loaded onto an Isolute^® SCX-2 cartridge which was flushed with MeOH.

The product was then eluted using 2M NH₃ in MeOH and evaporated to give

Intermediate 4 as an oil.

Yield: 44 mg (72%)

LC-MS (Method 2): Rt 1.95 min, m/z 436 [M+H]⁺

Intermediate 5

(fl)-4-(4.Cyanophenyl)-6-methyl-2-oxo-1-(3-trifluoromethylphenyl)-1,2,3,4-tetrahydro- pyrimidine-5-carboxylic acid (WO06/082412) (40.0 g, 0.010 mol) was dissolved in a mixture of MeOH (100 ml) and toluene (200 ml) and a 2M solution of (trimethylsilyl)diazomethane (100 ml, 0.2 mol) in hexanes was added dropwise. After stirring at RT for 30 min, the reaction mixture was evaporated to dryness to give Intermediate 5 as a cream solid. Yield: 40.4 g (95%)

LC-MS (Method 1): Rt = 3.68, m/z = 416 [M+H]⁺

Intermediate 6

A solution of Intermediate 5 (35.0 g, 0.09 mol) in chloroform (150 ml) was treated with bromine (14.87 g, 0.09 mmol) in chloroform (50 ml). The reaction mixture was stirred at RT for 30 min and then evaporated to give an orange foam. The product was purified by crystallization from EtOAc and pentane to give Intermediate 6 as a cream solid.

Yield: 38.04 g (91%)

LC-MS (Method 2): Rt = 3.66 min, m/z = 494/496 [M+H]⁺

Intermediate 7

Triethylamine (1.10 ml, 7.90 mmol) was added to 10-amino-1 -decanol (176 mg, 1.01 mmol) and Intermediate 6 (0.50 g, 1.01 mmol) in THF (10 ml) and the reaction mixture was stirred at RT under nitrogen for 20 h. The solvent was removed in vacuo, water (100 ml) was added to the residue and the product was extracted into EtOAc (2 x 50 ml). The extracts were washed with water (2 x 50 ml) and brine (50 ml) before the organic phase was isolated, dried (MgSO₄), filtered, and concentrated. The residue was triturated using Et₂O to afford Intermediate 7 as a white solid. Yield: 430 mg (76%)

LC-MS (Method 1): Rt 3.66 min, m/z 555 [M+H]⁺

Intermediate 8

Methanesulphonyl chloride (93 μl, 1.20 mmol) was added to a stirred solution of Intermediate 7 (415 mg, 0.75 mmol) in DCM (7 ml) at 0-5⁰C under nitrogen and the reaction was stirred for 1.5 h. The mixture was diluted with DCM (75 ml) then washed with brine (50 ml) before the organic phase was isolated, dried (MgSO₄), filtered and concentrated in vacuo to afford Intermediate 8 as a foam.

Yield: 0.46 g (98%)

LC-MS (Method 1 ): Rt 3.90 min, m/z 633 [M+H]⁺

Triethylamine (120 μl, 0.86 mmol) was added to a solution of Intermediate 6 (50 mg, 0.10 mmol) and Intermediate 4 (44 mg, 0.10 mmol) in THF (2 ml) at RT under nitrogen and the reaction mixture was stirred for 20 h. The solvent was removed in vacuo, water (25 ml) was added and the product was extracted into EtOAc (2 x 25 ml). The extracts were washed with water (2 x 25 ml) and brine (25 ml) before the organic phase was isolated, dried (MgSO₄), filtered and concentrated in vacuo. Purification by flash column chromatography using an Isolute® Si Il cartridge eluting with 50% EtOAc in cyclohexane, EtOAc then a MeOH/EtOAc gradient before preparative HPLC (system 1 ) and isolation of the free base using an Isolute® SCX-2 cartridge gave a residue that was freeze-dried to afford Example 1 as a white solid. Yield: 15 mg (18%) LC-MS (Method 3): Rt 8.75 min, m/z 817.40 [M+H]⁺

Example 2

DIPEA (120 μl, 0.690 mmol) was added to a solution of Intermediate 8 (105 mg, 0.166 mmol) and (5-methylaminomethyloxazol-2-yl)diphenylmethanol (WO2007/017669) (40 mg, 0.136 mmol) in acetonitrile (1 ml) and the reaction mixture was heated at 50°C in a sealed reaction tube for 48 h. The solvent was removed in vacuo and the residue purified using an Isolute® Si Il cartridge eluting with 50% EtOAc in cyclohexane, EtOAc then a MeOH/EtOAc gradient before preparative HPLC (system 1). Isolation of the free base using an Isolute® SCX-2 cartridge gave a residue that was freeze-dried to afford Example 2 as a white solid. Yield: 29 mg (25%) LC-MS (Method 3): Rt 9.00 min, m/z 831.38 [M+H]⁺

The following examples were prepared in a similar manner from Intermediate 8::.ι

Example 5

A 30% solution of bromomethane in acetonitrile (2.5 ml) was added to Example 2 (33 mg, 39.7 μmol) and the solution was heated at 40°C in a sealed reaction tube for 24 h. The solvent was removed under a stream of nitrogen and the residue purified using an AITN cartridge eluting with a 0-10% MeOH in DCM. The pure fractions were combined and evaporated to afford a residue that was freeze-dried to give Example 5 as a white solid. Yield: 27 mg (73%) LC-MS (Method 3): Rt 8.99 min, m/z 845.36 [M]⁺

The following examples were prepared in a similar manner:

* Purified using HPLC (system 2) Biological Assays

Compounds of the invention were tested for their HNE inhibitory activity and their M3 receptor binding activity in the following assays.

A. HNE Inhibition

Fluorescent peptide substrate

Assays were performed in 96-well plates at a total assay volume of 100 μl. The final concentration of the enzyme (human leukocyte elastase, Sigma E8140) was 0.00036 units/well. A peptide substrate (MeO-Suc-Ala-Ala-Pro-ValAMC, Calbiochem #324745) was used, at the final concentration of 100 μM. The final concentration of DMSO was 1% in the assay buffer (0.05M Tris.HCI, pH 7.5, 0.1 M NaCI; 0.1 M CaCI2; 0.0005% brij-35).

The enzymatic reaction was started by adding the enzyme. The enzymatic reaction was performed at RT and after 30mins stopped by adding 50 μl soybean trypsin inhibitor (Sigma T-9003) at a final concentration of 50μg/well. Fluorescence was read on the FLEXstation (Molecular Devices) using 380 nm excitation and 460 nm emission filters. The potency of the compounds was determined from a concentration series of 10 concentrations in range from 1000 nM to 0.051 nM. The results are means of two independent experiments, each performed in duplicate. Using Fluorescently labelled elastin

Assays were performed in 96-well plate at a total assay volume of 100 μl. The final concentration of the enzyme (human leukocyte elastase, Sigma E8140) was 0.002 units/well. Fluorescently labelled, solubilised elastin from bovine neck ligament (Molecular Probes, E-12056) was used at the final concentration of 15 μg/ml. The final concentration of DMSO was 2.5% in the assay buffer (0.1 M Tris- HCL, pH8.0, containing 0.2mM sodium azide).

The enzymatic reaction was started by adding the enzyme. The enzymatic reaction was performed at RT and read after 120 minutes. Fluorescence was read on the FLEXstation (Molecular Devices) using 485 nm excitation and 530 nm emission filters. The potency of the compounds was determined from a concentration series of 10 concentrations in range from 25000 nM to 1 nM. The results are means of two independent experiments, each performed in duplicate. All compounds of the Examples had activities in the range IC₅₀ = 1 -100 nM. HNE induced lung haemorrhage in the rat

Instillation of human neutrophil elastase (HNE) into rat lung causes acute lung damage. The extent of this injury can be assessed by measuring lung haemorrhage. Male Sprague Dawley rats (175-22Og) were obtained from Harlan UK Ltd., full barrier-bred and certified free from specified micro-organisms on receipt. Animals were weighed and randomly assigned to treatment groups (7-12 animals per group).

The vehicle used was 1% DMSO/Saline. Inhibitors were dissolved in 1%

DMSO before the addition of 0.9% saline. Animals in each study used to determine the efficacy of the elastase inhibitors delivered locally to the lung by a variety of routes. Rats were anaesthetised with the inhaled anaesthetic lsoflurane (4%) when the dose was given from 30 minutes to 6h . prior to human neutrophil elastase (HNE) administration or terminally anaesthetised with hypnorm:hypnovel:water (1.5:1 :2 at 2.7 ml/kg) when the predose was given at less than 30 minutes prior to HNE administration and dosed either intratracheal^ (i.t.) by transoral administration using a Penn Century microsprayer or intranasally (i.n.) by dropping the fluid on to the nares. Animals either received vehicle or compound at a dose volume of 0.5 ml/kg.

Animals that had been allowed to recover after dosing were terminally anaesthetised with hypnorm.hypnovel.water (1.5:1 :2 at 2.7ml/kg). Once sufficiently anaesthetised, HNE (600units/ml) or sterile saline was administered by transoral tracheal instillation at a volume of 100 μl using a Penn Century microsprayer. Animals were kept warm in a temperature controlled box and given top up doses of anaesthetic as required to ensure continuous anaesthesia until termination. Animals were sacrificed (0.5ml to 1 ml sodium pentobarbitone) one hour post

HNE challenge. The trachea was exposed and a small incision made between two tracheal rings allowing a cannula (1 Ogauge, O.D.2-10mm, Portex Ltd.) to be inserted approximately 2cm into the trachea towards the lung. This was secured into place with a cotton ligature. The lungs were then lavaged (BAL) three times with fresh 4ml aliquots of heparinised (1 Ounits/ml) phosphate buffered saline (PBS). The resultant BALF was kept on ice until it was centrifuged.

The BALF was centrifuged at 1000 r.p.m. for 10 minutes in a centrifuge cooled to between 4 and 10⁰C. The supernatant was discarded and the cell pellet resuspended in 1ml 0.1% CETAB/PBS to lyse the cells. Cell lysates were frozen until spectrophotometric analysis for blood content could be made. Standards were prepared by making solutions of whole rat blood in 0.1% CETAB/PBS.

Once defrosted 100 μl of each lysed cell suspension was placed into a separate well of a 96 well flat bottomed plate. All samples were tested in duplicate and 100 μl 0.1 % CETAB/PBS was included on the plate as a blank. The OD of the contents of each well was measured at 415nm using a spectramax 250 (Molecular devices).

A standard curve was constructed by measuring the OD (at 415 nm) of different concentrations of blood in 0.1 % CETAB/PBS (30, 10, 7, 3, 1 , 0.3, 0.1 μl/ml). The amount of blood in each experimental sample was calculated by comparison to the standard curve. Data were then analysed as below:

1 ) The mean OD for duplicates was calculated

2) The value for the blank was subtracted from the value for all other samples 3) Data were assessed to evaluate the normality of distribution.

B. Muscarinic Receptor Radioligand Binding Assays

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

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

The rate at which [3H]-QNB is detected binding to the muscarinic receptors is related to the rate at which the antagonist dissociates from the receptor, i.e. to the half life of the antagonists on the receptors.

All compounds of the Examples were shown to bind (IC₅₀ < 500 nM) to the M3 receptor. Analysis of Inhibition of M3 Receptor Activation via Calcium Mobilization

CHO cells expressing the human M3 receptor were seeded and incubated overnight in 96 well collagen coated plates (black-wall, clear bottom) at a density of 50000 / 75 μl of medium in 3% serum. The following day, a calcium-sensitive dye (Molecular Devices, Cat # R8041 ) was prepared in HBSS buffer with the addition of 5mM probenecid (pH 7.4). An equal volume of the dye solution (75 μl) was added to the cells and incubated for 45 minutes followed by addition of 50 μl of muscarinic antagonists or vehicle. After a further 15 minutes the plate was read on a FLEXstation™ (excitation 488nm, emission 525nm) for 15 seconds to determine baseline fluorescence. The muscarinic agonist Carbachol was then added at an EC_8O concentration and the fluorescence measured for a further 60 seconds. The signal was calculated by subtracting the peak response from the mean of the baseline fluorescence in control wells in the absence of antagonist. The percentage of the maximum response in the presence of antagonist was then calculated in order to generate IC₅₀ curves.

Claims

Claims:

1. A compound of formula (IA) or (IB):

wherein A is aryl or heteroaryl;

D is oxygen or sulphur;

R¹, R², R³ and R⁵ are independently each hydrogen, halogen, nitro, cyano, C|-C₆-alkyl, C₂-C₆-alkenyl, C₂-C₆-alkynyl, hydroxy or CrCe-alkoxy or C₂-C₆- alkenyloxy,, wherein CrCε-alkyl and C_rC₆-alkoxy can be further substituted with one to three identical or different radicals selected from the group consisting of halogen, hydroxy and C_rC₄-alkoxy;

R⁴ is hydrogen, Ci-C₆-alkyl, formyl, aminocarbonyl, mono- or di-CrC₄- alkylaminocarbonyl, C₃-C₈-cycloalkylcarbonyl, d-Cβ-alkylcarbonyl, Ci-C₆- alkoxycarbonyl, N-(Ci-C₄-alkylsulfonyl)-aminocarbonyl, N-CCrC_A-alkylsulfonylJ-N-^r C₄-alkyl)-aminocarbonyl, heteroaryl, heterocycloalkyl, heteroarylcarbonyl or heterocycloalkylcarbonyl; wherein Ci-C₆-alkyl, mono- and Ui-C₁ -C₄- alkylaminocarbonyl, Ci-C₆-alkylcarbonyl, CrC₆-alkoxycarbonyl, heteroaryl and heterocycloalkyl can be substituted with one to three identical or different radicals selected from the group consisting of aryl, heteroaryl, hydroxyl, CrC₄-alkoxy, hydroxycarbonyl, Ci-C₆-alkoxycarbonyl, aminocarbonyl, mono and Ui-C₁-C₄- alkylaminocarbonyl, amino, mono- and di-Ci-C₄-alkylamino, C₁-C₄- alkylcarbonylamino, cyano, N-(mono- and di-C₁-C₄-alkylamino-Ci-C₄-alkyl)- aminocarbonyl, N-(Ci-C₄-alkoxy-Ci-C₄-alkyl)-aminocarbonyl or halogen; -[Linker]- is a divalent linker radical; and M is a moiety having M3 receptor antagonist activity.

2. A compound as claimed in claim 1 , wherein the -[Linker]- radical is a divalent straight chain, saturated or unsaturated hydrocarbon radical having from 2 to 12 carbon atoms in the said chain, and wherein one or more carbons may be replaced by a divalent monocyclic or bicyclic carbocyclic or heterocyclic radical having from 3 to 7 ring atoms in the or each ring, or by -O-, -S-, -S(=O)-, -S(=O)₂-, -C(=O)-, -N(R^P)-, -N⁺(R^P)(R^Q)-, -C(=O)O-, -OC(=O)-, -C(=O)NR^A -, -NR^AC(=O)-, -S(O₂)NR^A-, -NR^AS(O₂)-, -NR^AC(=O)NR^B-, -NR^AC(=NR^A)NR^B-, -C(=NR^D)NR^E-, or -NR^EC(=NR^D)-, wherein R^A, R^B, R^D and R^E are independently hydrogen, C₁-C₆ alkyl, or C₃-C₆ cycloalkyl, and R^p and R^Q are independently hydrogen, C₁-C₆ alkyl, or C₃-C₆ cycloalkyl, HO-(C₁-C₆ alkyl)-, R^AR^BN-(C_rC₆ alkyl)-, or HOCf=O)-(C₁-C₆ alkyl)-, or R^Aand R^B, or R^D and R^E, or R^p and R^Q taken together with the nitrogens to which they are attached form a monocyclic heterocyclic ring of 5 to 7 ring atoms which may contain a further heteroatom selected from N, O and S.

3. A compound as claimed in claim 2 wherein when one or more one or more -(CH₂)- groups of the -[Linker]- radical is or are replaced by a divalent monocyclic or bicyclic carbocyclic or heterocyclic radical, the said radical is selected from the following:

4. A compound as claimed in claim 2 wherein -[Linker]- radical has one of the following structures (A), (B)₁(C), (D) and (E): (CH₂)₂.₅-N(CH₃)-(CH₂)₂.₅- (A)

(CH₂)₂.₅-N⁺(CH₃)₂-(CH₂)₂.₅ (B)

(CH₂)₂.₅ NH-(C=NH)-NH (CH₂)₂.₅ ^■ (C)

(CH₂),₅ (CH₂)₂, _ - (CH₂ (CH₂W

(D) _(E)

5. A compound as claimed in claim 2 wherein the -[Linker]- radical has one of the following structures (G) and (E):

(CHaJ^-CO-NH-tCHaJ^-N⁺tCH^CH^^-NH-CCMCHa)^— (H)

6. A compound as claimed in claim 1 wherein -[Linker]- is a divalent radical of formula -[X]_m-[Alk¹ ]_P-[Q]_n-[Alk²]_q-[X¹]_k- wherein k, m, n, p and q are independently 0 or 1 ;

AIk¹ and AIk² each independently represent an optionally substituted CrC₆ alkylene, or C2-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-) or amino (-NR^A-) link wherein R^A is hydrogen or CrC₃ alkyl;

X² represents (i) -O-, -S-, -S(=O)-, -Sf=O)₂-, -S⁺(R^A)-, -N(R^A)-, -N⁺(R^A)(R^B)-, -C(=O)-, -C(=O)O-, -OC(=O)-, -C(=O)NR^A -, -NR^AC(=O)-, -S(O₂)NR^A-, -NR^AS(O₂)-, -NR^AC(=O)NR^B-, -NR^AC(=NR^A)NR^B-, -C(=NR^D)NR^E-, -NR^EC(=NR^D)-, wherein R^A, R^B, R^D and R^E are independently hydrogen, CrC₆ alkyl, or C₃-C₆ cycloalkyl, or R^A and R^R or R^D and R^E taken together with the nitrogen to which they are attached form a monocyclic heterocyclic ring of 5 to 7 ring atoms which my contain a further heteroatom selected from N, O and S, or (ii) an optionally substituted divalent mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members; X represents -(C=O)-, -S(O₂)-, -C(=O)O-, -(C=O)NR^A-, or -S(O₂)NR^A-, wherein R^A is hydrogen, CrC₆ alkyl, or C₃-C₆ cycloalkyl; X¹ represents -O-, -S-, or -NH; and

Q is a divalent radical which contains an anionic-cationic pair selected from (i) a negatively charged nitrogen and positively charged nitrogen, (ii) a negatively charged oxygen and a positively charged nitrogen or (iii) a negatively charged nitrogen and a positively charged sulfur or (iv) a negatively charged oxygen and a positively charged sulfur. . .

7. A compound as claimed in claim 6 wherein Q represents a divalent radical of formula:

-S⁺(A-Qⁱ-(B)₅-Z¹)-, -(R^B)N⁺(A-Q¹-(B)_S-Z¹)- or X N/⁺Tλ H-(A-Q¹-(B)_S-Z¹),

wherein s is 0 or 1 ; ring L represents a mono- or bicyclic ring or ring system having 3 to 6 ring atoms in the case of a monocyclic ring and up to 10 ring atoms in the case of a bicyclic ring system, A and B each independently represent an optionally substituted divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic radical, or an optionally substituted C₁-C₆ alkylene or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic link wherein R^A is hydrogen Ci-C₃ alkyl or aryl(Ci-C₃ alkyl)-; Q¹ is an anionic divalent radical selected from (1) to (11):

R^B is hydrogen, C₁-C₆ alkyl, C₃-C₆ cycloalkyl or aryl(CrC₃ alkyl)-; and Z¹ is hydrogen, trifluoromethyl, or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members

8. A compound as claimed in claim 6 wherein Q represents a divalent radical of formula:

wherein ring T represents a mono- or bicyclic ring or ring system having 3 to 6 ring atoms in the case of a monocyclic ring and up to 10 ring atoms in the case of a bicyclic ring system, and Q¹ is an anionic divalent radical selected from those of formulae (1) to (11) as defined in claim 7.

9. A compound as claimed in claim 6 wherein Q represents a divalent radical of formula:

-S⁺(A-Z¹)-, -N⁺(A-Z¹ )(R^B)- or N⁺ -(A-Z¹)

V^ wherein ring L is as defined in claim 3, A represents an optionally substituted divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic radical, or an optionally substituted CrC₆ alkylene or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic link wherein R^A is hydrogen, C₁-C₃ alkyl or aryl(C_rC₃ alkyl)-; R^B is hydrogen, C₁-C₆ alkyl, aryl(C_rC₃ alkyl)-, or C₃-C₆ cycloalkyl; and Z¹ is hydrogen, trifluoromethyl, or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members, and wherein radical -A-Z¹ is substituted by an anionic group selected from those of formulae (12) to (20):

5) (16)

or by an anionic group selected from formulae (1) to (11 ) as defined in claim 3 but wherein one unsatisfied valency is satisfied by a group -A-Z¹.

10. A compound as claimed in claim 6 wherein Q represents a divalent radical -N (E-A-Q¹ -B-Q²-(D)_S-Z¹)- or -(R^B)C(E-A-Q¹-B-Q²-(D)_S-Z¹)- wherein E is a bond, -C(=O)-, or -S(=O)₂-; s is 0 or 1 ; A, B and D independently represent an optionally substituted divalent monocyclic 3- to 6-membered carbocyclic or heterocyclic radical, or an optionally substituted C₁-C₆ alkylene, or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6-membered carbocyclic or heterocyclic link wherein R^A is hydrogen, C₁-C₃ alkyl or aryl(CrC₃ alkyl)-; Z¹ is hydrogen, trifluoromethyl, or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members; one of Q¹ and Q₂ is an anionic divalent radical selected from those of formulae (1 ) to (11 ) as defined in claim 7, while the other of Q¹ and Q² is a divalent cationic radical selected from those of formulae (20) to (26):

wherein ring K is a 4- to 8-membered saturated ring which may contain 1 or 2 ^s- additional heteroatoms N, O or S; ring J is a 5- or 6-membered heteroaromatic rung; R' and R" are each hydrogen, or an optional substituent, or R' and R" taken together represent a divalent C₁-C₆ alkylene or C₂-C₆ akenylene radical in which up to three carbon atoms may be replaced by N O or S atoms.

11. A compound as claimed in claim 6 wherein Q represents a divalent radical of formula -N(E-A-Q¹-B-Z¹)- or -(R^B)C(E-A-Q¹-B-Z¹)-, wherein E is a bond,

-C(=O)-, or -S(=O)₂-; A and B independently represent an optionally substituted divalent monocyclic 3- to 6-membered carbocyclic or heterocyclic radical, or an optionally substituted d-C₆ alkylene, or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6-membered carbocyclic or heterocyclic link wherein R^A is hydrogen, Ci-C₃ alkyl or aryl(Ci-C₃ alkyl)-; Z¹ is hydrogen, trifluoromethyl, or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members; radical -B-Z¹ is substituted by Q²; Q¹ is selected from divalent anionic radicals (1) to (11) as defined in claim 7 and Q² is selected from cationic radicals (12) to (20) as defined in claim 9, OR Q¹ is selected from divalent cationic radicals (21) to (27) as defined in claim 10 and Q² is selected from anionic groups of formulae (12) to (20) as defined in claim 9 or anionic groups selected from formulae (1 ) to (11 ) as defined in claim 7 but wherein one unsatisfied valency is satisfied by a group -A-Z¹.

12. A compound as claimed in claim 6 wherein Q represents -(R^B)C(A-Z¹)- wherein A represents an optionally substituted divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic radical, or an optionally substituted CrC₆ alkylene, or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic link wherein R^A is hydrogen or Ci-C₃ alkyl; R^B is hydrogen, CrC₆ alkyl, or C₃-C₆ cycloalkyl; Z¹ is hydrogen, trifluoromethyl or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members; and wherein radical -A-Z¹ is substituted by a group selected from:

<²⁸> (29) wherein R' and R" are each hydrogen, CrC₃ alkyl or aryl(CrC₃ alkyl)-, or R' and R" taken together with the nitrogen to which they are attached form a 3- to 8-membered heterocyclic ring, and wherein R'" represents hydrogen or one or more optional substituents.

13. A compound as claimed in claim 6 wherein Q represents a divalent radical of formula -(Z²-(B¹)_r-Q²-A¹)C(A-Q¹ -(B)₃-Z¹)- wherein r and s are independently 0 or 1 ; A, A¹, B and B¹ each independently represent an optionally substituted divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic radical, or an optionally substituted CrC₆ alkylene, or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic link wherein R^A is hydrogen, C₁-C₃ alkyl or 8171(C₁-C₃ alkyl)-; Z¹ and Z² independently represent hydrogen, trifluoromethyl, or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members; one of Q¹ and Q² is an anionic divalent radical selected from those of formulae (1 ) to (1 1 ) as defined in claim 7 while the other of Q¹ and Q² is a cationic divalent radical selected from those of formulae (21) to (27) as defined in claim 10.

14. A compound as claimed in claim 6 wherein Q represents a divalent radical of formula -(Z²-A¹)C(A-Q¹-(B)_S-Z¹)- wherein r and s are independently 0 or 1 ; A, A¹, and B each independently represent an optionally substituted divalent monocyclic 3- to 6- membered carbocyclic or heterocyclic radical, or an optionally substituted CrC₆ alkylene, or C₂-C₆ alkenylene radical which may optionally contain an ether (-O-), thioether (-S-), amino (-NR^A-) or divalent monocyclic 3- to 6-membered carbocyclic or heterocyclic link wherein R^A is hydrogen, Ci-C₃ alkyl or aryl(CrC₃ alkyl)-; Z¹ and Z² independently represent hydrogen, trifluoromethyl, or an optionally substituted mono- or bicyclic carbocyclic or heterocyclic radical having 3-6 ring members; radical Z²-A¹- is substituted by Q²; and radical -B-Z¹ is substituted by Q²; Q¹ is selected from divalent anionic radicals (1) to (11) as defined in claim 7 and Q² is selected from cationic groups (12) to (20) as defined in claim 9, OR Q¹ is selected from divalent cationic radicals (21 ) to (27) as defined in claim 10 and Q² is selected from anionic groups of formulae (12) to (20) as defined in claim 9 or groups of formulae (1 ) to (11) as defined in claim 7 but wherein one unsatisfied valency is satisfied by a group -A-Z¹.

15. A compound as claimed in claim 1 or claim 2 wherein Q represents a divalent radical selected from:

(30) (3D wherein R' is hydrogen, CrC₃ alkyl or aryl(CrC₃ alkyl)-

16. A compound as claimed in any of the preceding claims wherein R⁴ is hydrogen.

17. A compound as claimed in any of the preceding claims wherein R¹, R² and R³ are independently each hydrogen, halogen, nitro, cyano, Cι-C₃-alkyl, C₂-C₃-alkenyl, C₂-C₃-alkynyl, hydroxy or d-C₃-alkoxy or C₂-C₃-alkenyloxy.

18. A compound as claimed in any of claims 1 to 16 wherein R¹, R² and R³ are independently each hydrogen, fluoro, chloro, bromo, cyano, methyl, methoxy and

-C≡CH.

19. A compound as claimed in any of the preceding claims wherein A is phenyl, pyridyl, or pyrimidinyl.

20. A compound as claimed in any of the preceding claims wherein one of R¹ and R² is methyl, -C≡CH or cyano.

21. A compound as claimed in any of the preceding claims wherein -AR¹R² is 4-cyanophenyl or 4-ethynylphenyl.

22. A compound as claimed in any of the preceding claims wherein D is O.

23. A compound as claimed in any of the preceding claims wherein R⁵ is hydrogen and R³ is 3-trifluoromethyl, 3-chloro or 3-bromo.

24. A compound as claimed in any of the preceding claims wherein Z is a radical of formula (IA') or (IB'):

(IA') (IB¹)

25. A compound of formula (X):

wherein m, n, p, q, k, X, X¹, AIk¹ and AIk² are as defined in claim 1 , and Q is as defined in any of claims 1 to 15.

26. A compound as claimed in any of the preceding claims wherein m and k are each 0, p and q are each 1 , and AIk¹ and AIk² are each -(CH₂W-

27. An ester of any compound claimed in any of the preceding claims which has a carboxylate, sulfonate or phosphonate group in the radical Q.

28. An ester as claimed in claim 27 which is a C₁-C₆ alkyl ester.

29. A compound as claimed in any of the preceding claims in pharmaceutically acceptable salt form.

30. A pharmaceutical composition comprising a compound as claimed in any of claims 1 to 29 and a pharmaceutically acceptable carrier or excipient.

31. A pharmaceutical composition as claimed in claim 30 which is adapted for administration by the pulmonary route.

32. Use of a compound as claimed in any of claims 1 to 29, for the manufacture of a medicament for use in the treatment of prevention of a disease or condition in which HNE activity and/or M3 receptor stimulation activity is implicated.

33. A method of treatment of a disease or condition in which HNE activity and/or M3 receptor stimulation activity is implicated, comprising administering to a.subject suffering such disease an effective amount of a compound as claimed in any of claims 1 to 29.

34. Use according to claim 32, or a method of treatment according to claim 33, wherein the disease or condition is chronic obstructive pulmonary disease (COPD), chronic bronchitis, lung fibrosis, pneumonia, acute respiratory distress syndrome (ARDS), pulmonary emphysema, smoking-induced emphysema or cystic fibrosis.

35. Use according to claim 29, or a method of treatment according to claim 33, wherein the disease or condition is asthma, rhinitis, psoriasis, dermatitis, (atopic and non-atopic), Crohn's disease, ulcerative colitis, or irritable bowel disease.