WO2008149110A1

WO2008149110A1 - Bicyclor [2.2.1] hept-7-ylamine derivatives and their use in the treatment of diseases and conditions in which m3 muscarinic receptor activity and beta-adrenergic activity are implicated

Info

Publication number: WO2008149110A1
Application number: PCT/GB2008/001955
Authority: WO
Inventors: Lilian Alcaraz; Nicholas Kindon; Jonathan Mark Sutton
Original assignee: Argenta Discovery Limited; Astrazeneca Ab
Priority date: 2007-06-08
Filing date: 2008-06-06
Publication date: 2008-12-11

Abstract

The present invention relates to compounds of formula (I) having muscarinic M3 receptor and β-adrenergic receptor modulating activity; wherein R1, R2, R3 and X are as defined herein.

Description

BICYCLOR [2.2.1] HEPT-7-YLAMINE DERIVATIVES AND THEIR USE IN THE TREATMENTOF DISEASES AND CONDITIONS IN WHICH M3 MUSCARINIC RECEPTOR ACTIVITY AND BETA-ADRENERGIC ACTIVITY ARE IMPLICATED

Field of the Invention

This invention relates to bicyclo[2.2.1]hept-7-ylamine derivatives, pharmaceutical 5 compositions, methods for their preparation and use in the treatment of M3 muscarinic receptor mediated diseases, for example respiratory diseases.

Background to the invention

Anticholinergic 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 5 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. 0 Ther., 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 5 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 0 variety of different pathophysiological states including asthma and COPD. In COPD, vagal tone may either be increased (Gross et al. 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 et al. 1984, Am Rev Respir Dis; 129:856-870). In addition, inflammatory conditions can lead to a loss of inhibitory 5 M2 receptor activity which results in increased levels of acetylcholine release following vagal nerve stimulation (Fryer et al, 1999, Life ScL, 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.

Despite the large body of evidence supporting the use of anti-muscarinic receptor therapy for treatment of airway disease states, relatively few anti-muscarinic compounds are in use in the clinic for pulmonary indications. Thus, there remains a need for novel compounds that are capable of causing blockade at M3 muscarinic receptors, especially those compounds with a long duration of action, enabling a once-daily dosing regimen. Since muscarinic receptors are widely distributed throughout the body, the ability to deliver anticholinergic drugs directly to the respiratory tract is advantageous as it allows lower doses of the drug to be administered. The design and use of topically active drugs with a long duration of action and that are retained on the receptor or in the lung would allow reduction of unwanted side effects that could be seen with systemic administration of the same drugs.

Tiotropium (Spiriva ™) is a long-acting muscarinic antagonist currently marketed for the treatment of chronic obstructive pulmonary disease, administered by the inhaled route.

Tiotropium

Additionally ipratropium is a muscarinic antagonist marketed for the treatment of COPD.

Ipratropium

Other muscarinic receptor modulators have been referred to. For example: US4353922 describes muscarinic modulators based upon a [2.2.1]azabicycloheptane ring system. EP418716 and US005610163 describe various [3.2.1 Jazabicyclooctane ring systems. WO06/017768 describes [3.3.1]azabicyclononane ring systems. [2.2.2]azabicyclooctane systems (quinuclidines) have been previously described, for example in US2005/0209272 and WO06/048225. [3.1.0]azabicyclohexane systems have been described in, for example in WO06/035282. [3.2.1 Jazabicyclooctane systems have been described in for example WO06/035303.

The class of β2 adrenergic receptor agonists is well known. Many known β2-agonists, in particular, long-acting β2-agonists such as salmeterol and formoterol, have a role in the treatment of asthma and COPD. These compounds are also generally administered by inhalation. Compounds currently under evaluation as once-daily β2 agonists are described in Expert Opin. Investig. Drugs 14 (7), 775-783 (2005). A well known β2-agonist pharmacophore is the moiety:

Also known in the art are pharmaceutical compositions that contain both a muscarinic antagonist and a β2-agonist for use in the treatment of respiratory disorders. For example, US2005/0025718 describes a β2-agonist in combination with tiotropium, oxotropium, ipratropium and other muscarinic antagonists; WO02/060532 describes the combination of ipratropium with β2-agonists and WO02/060533 describes the combination of oxotropium with β2-agonists. Other M3 antagonist / β2-agonist combinations are described in WO04/105759 and WO03/087097. Also known in the art are compounds possessing both muscarinic receptor antagonist and β2-agonist activity present in the same molecule. Such bifunctional molecules provide bronchodilation through two separate modes of action whilst possessing single molecule pharmacokinetics. Such a molecule might be easier to formulate for therapeutic use as compared to two separate compounds and might be more easily co-formulated with a third active ingredient, for example a steroid. Such molecules are described in for example, WO04/074246, WO04/089892, WO05/111004, WO06/023457 and WO06/023460, all of which use different linker radicals for covalently linking the M3 antagonist to the β2-agonist.

Summary of the Invention

According to one aspect of the invention, there is provided a compound of formula (I):

wherein

X is OR⁴ or NR⁵R¹²;

(i) R¹ is d-Ce-alkyl, (C₃-C₆)cycloalkyl or hydrogen; and R² is a group-Z-NR⁹R¹⁰; and R³ is a lone pair, or C_rC₆-alkyl, in which cases 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 group -Z-NR⁹R¹⁰, 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 , -Z-NR⁹R¹⁰; and R³ is a lone pair, or Ci-C₆-alkyl, in which cases the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge;

R⁴ is selected from one of the groups of formula (a), (b), (c) or (d):

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

R⁵ is selected from one of the groups of formula (a), (b) or (c):

(a) (b) (C)

R⁶ is C_rC₆-alkyl, (C₃-C₆)cycloalkyl or a hydrogen atom;

R and R are a CrC₆-alkyl, (C₃-C₆)cycloalkyl group or halogen;

n and m are independently 0, 1 , 2 or 3;

each occurrence of R^8a and R^8b is independently selected from the group consisting of aryl, aryl-fused-heterocycloalkyl, heteroaryl, CVCValkyl, (C₃-C₆)cycloalkyl;

each occurrence of R^8c is independently selected from -OH, CVCValkyl, (C₃-

C₆)cycloalkyl, hydroxy-CrCβ-alkyl, nitrile, a group CONR^8d ₂ and a hydrogen atom;

R is d-C_δ-alkyl, (C₃-C₆)cycloalkyl or a hydrogen atom;

R is a hydrogen atom, (C₃-C₆)cycloalkyl or CrCe-alkyl;

R is an aryKCrCβ-alkyl)-, or heteroaryl(CrC₆-alkyl) group, wherein both CVC₆-alkyl species are optionally substituted by hydroxy;

R is CrC₆-alkyl, (C₃-C₆)cycloalkyl or a hydrogen atom;

Ar¹ is aryl, heteroaryl or (C₃-C₆)cycloalkyl; Ar² are, independently, aryl, heteroaryl or (C₃-C₆)cycloalkyl; and

Q is an oxygen atom, -CH₂-, -CH₂CH₂- or a bond;

Z is selected from:

(i) a C_rCi₆-alkylene, C₂-C₁₆-alkenylene or C₂-C₁₆-alkynylene group;

or

(ii) a linker ^*-C(L³)(L⁴)-L-C(L¹)(L²)-^** , wherein:

^* denotes the point of attachment to the non-aromatic nitrogen bearing R¹ and R³ or to the heterocycloalkyl ring formed by R¹ and R²; and ^** denotes the point of attachment to the group NR⁹R¹⁰; L is a hydrocarbyl chain of up to 14 carbon atoms, wherein one, two or three carbon atoms of the chain are replaced by groups independently selected from O, NR⁴⁵, S, S(O), S(O)₂, C(O)O, OC(O), NR⁴⁶C(O), C(O)NR⁴⁷, NR⁴⁸S(O)₂, S(O)₂NR⁴⁹, NR⁵⁰C(O)NR⁵¹, NR⁵²S(O)₂NR⁵³, OC(O)NR⁵⁴, NR⁵⁵C(O)O, provided that any heteroatoms in the chain are separated by at least 2 carbon atoms; and/or wherein up to four carbon atoms of the chain form part of a mono- or bicyclic aliphatic, heteroaliphatic, aromatic or heteroaromatic ring having up to three heteroatoms independently selected from N, O or S, said ring comprising up to 10 ring atoms, and wherein the ring is optionally substituted by up to three substituents independently selected from halogen, S(O)₀.₂R⁵⁶, NR⁵⁷R⁵⁸, S(O)₂NR⁵⁹R⁶⁰, C(O)NR⁶¹R⁶², C(O)OR⁶³, NR⁶⁴S(O)₂R⁶⁵, NR⁶⁶C(O)R⁶⁷, NR⁶⁸C(O)OR⁶⁹, NR⁷⁰C(O)NR⁷¹R⁷², OR⁷³, C_h alky! and C₃^₃ cycloalkyl, and wherein C_1-6 alkyl and C₃.₆ cycloalkyl may be optionally substituted by up to three substituents independently selected from halogen, hydroxyl and C_1-6 alkoxy; and the chain may comprise up to three of such rings each selected independently; wherein R⁵⁶, R⁶⁵ and R⁶⁹ each independently represent C_1-6 alkyl or C₃. 6 cycloalkyl, wherein C₁^ alkyl and C_3-6 cycloalkyl may be optionally substituted by up to three substituents independently selected from halogen, hydroxyl, Ci. 6 alkoxy; and R 45 _D46 _D47 p48 p49 p50 _D51 p52 p53 _D54 _D55 _D57 _D58 p59 p60 , n , π , n , n , n , π , n , π , ri , n , hi , π , ri , n ,

R⁶¹, R⁶², R⁶³, R⁶⁴, R⁶⁶, R⁶⁷, R⁶⁸ , R⁷⁰, R⁷¹, R⁷² and R⁷³ each independently represent hydrogen, or C_1-6 alkyl or C₃.₆ cycloalkyl, wherein C_1-6 alkyl and C₃.₆ cycloalkyl may be optionally substituted by up to three substituents independently selected from halogen, hydroxyl, C₁^ alkoxy; or any of R⁵⁷ and

R⁵⁸, R⁵⁹ and R⁶⁰, R⁶¹ and R⁶² or R⁷¹ and R⁷², together with the nitrogen atom to which they are both attached, may form a 4 to 8 membered aliphatic heterocyclic ring, wherein the aliphatic heterocyclic ring may comprise up to three heteroatoms independently selected from N₁O and S, wherein the ring may be optionally substituted by up to three substituents independently selected from halogen, hydroxyl and C_1-6 alkyl or C₃.₆ cycloalkyl, wherein C₁^ alkyl and C_3-6 cycloalkyl may be optionally substituted by one or more substituents independently selected from halogen and hydroxyl; and wherein the chain may additionally comprise up to three carbon- carbon double bonds; wherein the chain may additionally comprise up to three carbon- carbon triple bonds;

L¹ and L² each independently represent hydrogen, C₁^ alkyl or C₃.₆ cycloalkyl; L³ and L⁴ each independently represent hydrogen, C_1-6 alkyl or C₃.₆ cycloalkyl, wherein C_1-6 alkyl and C_3-6 cycloalkyl may be optionally substituted by one or more substituents independently selected from halogen and hydroxyl; in addition L¹ and/or L³ may be linked to carbon atoms of the hydrocarbyl chain in linker L to form aliphatic rings of up to 6 ring atoms, wherein each ring may comprise up to three heteroatoms independently selected from N, O and S;

with the proviso that when X is OR⁴ then Z is a linker ^*-C(L³)(L⁴)-L-C(L¹)(L²)-^**; or a pharmaceutically acceptable salt thereof.

In another aspect the present invention provides a prodrug of a compound of formula (I) as herein defined, or a pharmaceutically acceptable salt thereof.

In yet another aspect the present invention provides an N-oxide of a compound of formula (I) as herein defined, or a prodrug or pharmaceutically acceptable salt thereof.

In a further aspect the present invention provides a solvate (such as a hydrate) of a compound of formula (I) as herein defined, or an N-oxide, prodrug or pharmaceutically acceptable salt thereof.

In one subset of the compounds of the invention:

R¹ is d-C₆-alkyl ; R² is a group -Z-NR⁹R¹⁰ and R³ is a lone pair or C_rC₆-alkyl, in which cases the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge, or:

R¹ and R² together with the nitrogen to which they are attached represent a heterocycloalkyl ring, said ring being substituted by a group -Z-NR⁹R¹⁰ and R³ is a lone pair or Ci-C₆-alkyl, in which cases the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge; or: R¹ and R³ together with the nitrogen to which they are attached represent a heterocycloalkyl ring, and R² is a group -Z-NR⁹R¹⁰, in which cases the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge;

R⁴ is selected from one of the groups of formula (a), (b) or (c):

(a) (b) (C)

R⁶ is a hydrogen atom;

R j7a a _n„d_! o R7b are independently a Ci-C₆-alkyl group or halogen;

n and m are independently 0, 1 , 2 or 3;

R^8a and R^8b are independently selected from the group consisting of aryl, heteroaryl, CrCe-alkyl, (C₃-C₆)cycloalkyl;

R is -OH, CVCValkyl, hydroxy-CrCβ-alkyl, or a hydrogen atom; R⁹ is a hydrogen atom or CrC₆-a!kyl; or R⁹ is a hydrogen atom;

R¹⁰ is a group ArCH(OH)CH₂- or a group ArCH₂CH₂-; or R¹⁰ is a group ArCH(OH)CH₂-, wherein Ar is a group

wherein M¹ is S, C(O), NA⁵, CA⁶A⁷, CH₂CH₂, CH=CH, CH₂O or OCH₂;

M² is S, C(O), NA⁵, CA⁶A⁷, CH₂CH₂, CH=CH, CH₂O or OCH₂;

A¹, A², A³ and A⁴ are, independently, hydrogen, halogen, trifluoromethyl, cyano, carboxy, hydroxy, nitro, S(O)₂A⁸, NA⁹S(O)₂A¹⁰, C(O)NA¹¹A¹², NA¹³C(O)A¹⁴, C_1-6 alkyl, C_1-6 alkoxy, C(O)(CL₆ alkyl) or C(O)OC_1-6 alkyl; and A³ can also be CH₂OH, NHCHO, NHS(O)₂NA¹⁵A¹⁶ or NHSO₂A¹⁷;

A⁵, A⁶, A⁷, A⁹, A¹¹, A¹², A¹³, A¹⁴, A¹⁵ or A¹⁶ are, independently, hydrogen or C_1-6 alkyl; A⁸, A¹⁰ and A¹⁷ are, independently, Cm alkyl.

More conveniently Ar is selected from

wherein

M¹ is S, CH=CH, CH₂O or OCH₂;

M² is S, CH=CH, CH₂O or OCH₂;

A¹, A², and A⁴ are, independently, hydrogen, halogen, C_1-6 alkyl, C_1-6 alkoxy; and A³ can also be CH₂OH, NHCHO, NHS(O)₂NA¹⁵A¹⁶ or NHSO₂A¹⁷;

A or A are independently selected from hydrogen or C_1-6 alkyl;

A¹' is C_1-6 alkyl;

Examples of C_1-6 alkyl include C_1-4 alkyl and C_1-2 alkyl.

Examples of C_1-6 alkoxy include C_1-4 alkoxy and C_1-2 alkoxy.

More conveniently Ar is selected from:

wherein A¹ , A² and A⁴ are all hydrogen, A³ is CH₂OH, NHCHO and M¹ is CH=CH or S.

In a further subset of compounds of the invention:

X is OR⁴ or NR⁵R¹²;

R¹ is d-Cβ-alkyl ; R² is a group -Z-NR⁹R¹⁰ and R³ is a lone pair or C_rC₆-alkyl, in which cases the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge, or: R¹ and R² together with the nitrogen to which they are attached represent a heterocycloalkyl ring, said ring being substituted by a group -Z-NR⁹R¹⁰ and R³ is a lone pair or Ci-C₆-alkyl, in which cases the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge; or: R¹ and R³ together with the nitrogen to which they are attached represent a heterocycloalkyl ring, and R² is a group -Z-NR⁹R¹⁰, in which cases the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge; or alternatively:

R¹ is H; R² is -Z-NR⁹R¹⁰ & R³ is a lone pair;

R⁴ and R⁵ are independently selected from one of the groups of formula (a), (b) or (c):

(a) (b) (C)

In the group (a):

Ar¹ is a phenyl group; R⁶ is a hydrogen atom; n and m are 0;

In the group (b):

R^8a and R >8⁸b^b are both a phenyl group; or, R and R are both a thienyl group, or R' is a phenyl group and R is a cycloalkyl group, such as cyclopentyl or cyclohexyl;

R is -OH or CrC₆-alkyl (especially methyl or ethyl); In a further embodiment, R is -OH;

In the group (c):

Ar² is a phenyl ring

Q is an oxygen atom

R^8c is hydrogen, C_1-6 alkyl (especially methyl) or -OH R⁹ is a hydrogen atom or CrCg-alkyl;

R¹⁰ is an aryl(C_rC₆-alkyl)-, or heteroaryKd-Ce-alkyl) group, selected from the group

In some compounds of the invention, X is OR⁴ and the radical MD(L₁)(L₂J-L-C(L₃)(L₄)-

(i) -(CH₂)S-O-(CH₂)I-B-(CH₂)U-, wherein B is a phenylene radical or a 5- or 6- membered heteroarylene radical and s is 2 or 3, t is 0, 1 or 2, and u is 1 or 2; or

(ii) -(CH₂)v-C(O)NR⁴⁷-B-(CH₂)u-, wherein v is 1 or 2 and R⁴⁷ is preferably hydrogen, but may also be C^alkyl such as methyl or ethyl, or C₃.₆ cycloalkyl such as cyclopropyl, B is a phenylene radical or a 5- or 6-membered heteroarylene radical, and u is 1 or 2; or

(iii) -(CH₂)V-NR⁴⁷C(O)-B-(CH₂)U-, wherein v is 1 or 2, R⁴⁷ is as defined in case (ii), B is a phenylene radical or a 5- or 6-membered heteroarylene radical, and u is 1 or 2; or

(iv) -(CH₂)v-C(O)NR⁴⁷-(CH₂)w-; wherein v is 1 or 2 and w is 2-8 and R⁴⁷ is as defined in case (ii); or

(v) -(CH₂Jy-NR⁴⁷C(O)-(CH₂)W-; wherein y and w are 2-8 and R⁴⁷ is as defined in case

(ii); or

(vi) -(CH₂)s-O-(CH₂)v--C(O)NR⁴⁷-(CH₂)w-; wherein v is 1 or 2 and w is 2-8, s is 2 or 3 and R⁴⁷ is as defined in case (ii); or

(vii) -(CH₂)v-B¹-(CH₂)w-; wherein v is 1 or 2 and w is 2-8 and B¹ is a phenylene radical, or a 5- or 6- membered heteroarylene radical or a bicyclic 8-10 membered arylene or heteroarylene radical, or a biarylene radical or biheteroarylene radical; or (viii) -(CH₂)v-B²-NR⁴⁷C(O)-(CH₂)u-, wherein v is 1 or 2, R⁴⁷ is as defined in case (ii), B² is a phenylene radical, and u is 1 or 2; or

(ix) -(CH₂)V-S(O)₂-B-(CH₂)U-, wherein v is 1 or 2, B is a phenylene radical or a 5- or 6- membered heteroarylene radical, and u is 1 or 2; or (x) -(CH₂)v-C≡C-B²-(CH₂)u-, wherein v is 1 or 2, u is 1 or 2 and B² is a phenylene radical.

In some compounds of the invention, X is NR⁵R¹² and the radical MD(L₁)(L₂J-L-

C(L₃)(U)-" is:

(i) -(CH₂)s-O-(CH₂)t-B-(CH₂)u-, wherein B is a phenylene radical and s is 2 or 3, t is 0,

1 or 2, and u is 1 or 2; or

(ii) -(CH₂)v-C(O)NR⁴⁷-B-(CH₂)u-, wherein v is 1 or 2 and R⁴⁷ is preferably hydrogen, but may also be C^alkyl such as methyl or ethyl, or C_3-6 cycloalkyl such as cyclopropyl, B is a phenylene radical, and u is 1 or 2; or

(iii) -(CH₂)V-NR⁴⁷C(O)-B-(CH₂)U-, wherein u, v, B and R⁴⁷ are as defined in case (ii) or

(iv) -(CH₂)v-C(O)NR⁴⁷-(CH₂)w-; wherein w is 3-8 and v and R⁴⁷ are as defined in case

(ii).

Compounds of the invention exist in either the syn- or anti- forms;

syn- anti-

Compounds of the invention also exist with the group -NR¹R²R³ in either the exo or endo orientation;

Currently it is preferred that the compounds of the invention be predominantly in the anti-endo configuration.

Compounds of the invention can also exist as optical isomers since substituted bicyclic ring systems can lack a plane of symmetry. The absolute configuration of the molecule can be defined using Cahn-lngold-Prelog rules to assign the R or S designation to each position. To avoid confusion the ring numbering used below is employed.

However, compounds of the invention include racemates, single enantiomers and mixtures of the enantiomers in any ratio, since all such forms have muscarinic M3 receptor modulating activity to varying extents.

The present invention relates to compounds wherein R⁴ is selected from one of the groups of formula (a), (b), (c) or (d):

(a) (b) (C) (d) and R⁵ is selected from one of the groups of formula (a), (b) or (c):

(a) (b) (C)

In the group (a), R⁶ may be C_rC₆-alkyl such as methyl or ethyl or a hydrogen atom; Ar¹ may be an aryl group such as phenyl, a heteroaryl group such as thienyl, especially 2-thienyl, or a cycloalkyl group such as cyclohexyl, cyclopentyl, cyclopropyl, or cyclobutyl; ring substituents R^7a and R^7b may be independently a d- C₆-alkyl group, such as methyl, ethyl, n- or isopropyl, n-, sec- or tertbutyl, or halogen such as fluoro, chloro or bromo; and m and n may be independently 0, 1 , 2 or 3.

In the groups (b) and (d), R^8a and R^8b may be independently selected from any of those aryl, aryl-fused-heterocycloalkyl, aryl-fused-cycloalkyl, heteroaryl, Ci-C₆-alkyl, or cycloalkyl groups specifically mentioned in the discussion of R⁵ above. Additionally, R^8b may also be a hydrogen atom. R^8c may be -OH, a hydrogen atom, CrC₆-alkyl such as methyl or ethyl, or hydroxy-CrCValkyl such as hydroxymethyl,. Presently preferred is the case where R^8c is -OH. Preferred combinations of R^8a and R^8b, especially when R^8c is -OH, include those wherein (i) each of R^8a and R^8b is optionally substituted monocyclic heteroaryl of 5 or 6 ring atoms such as pyridyl, oxazolyl, thiazolyl, furyl and especially thienyl such a 2-thienyl; (ii) R^8a and R^8b are both optionally substituted phenyl; (iii) one of R^8a and R^8b is optionally substituted phenyl and the other is cycloalkyl such as cyclopropyl, cyclobutyl, cycloheptyl, cyclooctyl or especially cyclopentyl or cyclohexyl; and (iv) one of R^8a and R^8b is optionally substituted monocyclic heteroaryl of 5 or 6 ring atoms such as pyridyl, thienyl, oxazolyl, thiazolyl, or furyl; and the other is cycloalkyl such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl.

In the group (c), R^8c may be -OH, a hydrogen atom, CrC₆-alkyl such as methyl or ethyl, or hydroxy-CrC₆-alkyl such as hydroxymethyl. Presently preferred is the case where R^8c is -OH. Each Ar² is an aryl, heteroaryl or cycloalkyl ring and may be, for example, any of those aryl, heteroaryl, C_rC₆-alkyl, or cycloalkyl rings specifically mentioned in the discussion of R⁵ above. Preferred Ar² rings include phenyl. The bridge -Q between the two Ar² rings is -O-, -CH₂-, -CH₂CH₂- or a bond.

Of the R⁴ options (a), (b), (c) and (d), it is presently preferred that R⁴ be a group (a) or (b) or (c).

One embodiment of the present invention provides compounds of formula (I) wherein the nitrogen attached to R¹ and R³ is a tertiary nitrogen. Typically in this embodiment, R¹ is H; R² is -Z-NR⁹R¹⁰ & R³ is a lone pair, wherein R⁹ and R¹⁰ are as previously defined.

One embodiment of the present invention provides compounds of formula (I) wherein the nitrogen attached to R¹ and R³ is a quaternary nitrogen and carries a positive charge.

One embodiment of the present invention provides compounds of formula (I) wherein X is NR⁵R¹² and Z is a CrC^-alkylene, C₂-C₁₆-alkenylene or C₂-C₁₆-alkynylene group.

One embodiment of the present invention provides compounds of formula (I) wherein X is NR⁵R¹² and Z is a linker ^*-C(L³)(L⁴)-L-C(L¹)(L²)-^**, wherein L, L¹, L², L³ and L⁴ are as previously defined.

One embodiment of the present invention proviides compounds of formula (I) wherein X is OR⁴.

One embodiment of the present invention provides compounds of formula (I) wherein: (i) R¹ is CrCValkyl or hydrogen; and R² is a group-Z-NR⁹R¹⁰; and R³ is a lone pair, or Ci-C₆-alkyl, in which cases 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 group -Z-NR⁹R¹⁰, 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 , -Z-NR⁹R¹⁰; and R³ is a lone pair or CrC₆-alkyl, in which cases the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge.

One embodiment of the present invention consists of compounds of formula (I) wherein R⁶ is C_rC₆-alkyl or a hydrogen atom.

One embodiment of the present invention consists of compounds of formula (I) wherein R^7a and R^7b are a C₁-CVaIkVl group or halogen.

One embodiment of the present invention consists of compounds of formula (I) wherein R^8c is -OH, CVCe-alkyl, hydroxy-C_rC₆-alkyl, nitrile, a group CONR^8d ₂ or a hydrogen atom. One embodiment of the present invention consists of compounds of formula (I) wherein R^8d is CrC₆-alkyl or a hydrogen atom.

One embodiment of the present invention consists of compounds of formula (I) wherein R⁹ is a hydrogen atom or C_rC₆-alkyl. One embodiment of the present invention consists of compounds of formula (I) wherein R⁹ is a hydrogen atom.

One embodiment of the present invention consists of compounds of formula (I) wherein R¹⁰ is a group ArCH(OH)CH₂-. One embodiment of the present invention consists of compounds of formula (I) wherein R¹⁰ is selected from the group:

One embodiment of the present invention consists of compounds of formula (I) wherein R¹² is CrC₆-alkyl or a hydrogen atom.

One embodiment of the present invention consists of a compound of formula (I) selected from:

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[3-(4-{[(R)-2-hydroxy- 2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}-phenoxy)- propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester;

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[3-(4-{[(R)-2-hydroxy- 2-(8-hydroxy-2-oxo-1 ,2-dihydro-quinolin-5-yl)-ethylamino]-methyl}-phenoxy)-propyl]- methyl-amino}-bicyclo[2.2.1]hept-7-yl ester;

9-Hydroxy-9H-xanthene-9-carboxylic acid (1 S,2S,4R,7R)-2-{[3-(4-{[(R)-2- hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}- phenoxy)-propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester;

9-Hydroxy-9H-xanthene-9-carboxylic acid (1 S,2S,4R,7R)-2-{[3-(4-{[(R)-2- hydroxy-2-(8-hydroxy-2-oxo-1 ,2-dihydro-quinolin-5-yl)-ethylamino]-methyl}-phenoxy)- propyl]-methyl-amino}-bicyclo[2.2.1 ]hept-7-yl ester; θ-Hydroxy-ΘH-xanthene-θ-carboxylic acid (1 S,2S,4R,7R)-2-{[3-(4-{[(R)-2- hydroxy-2-(5-hydroxy-3-oxo-3,4-dihydro-2/-/-benzo[1 ,4]oxazin-8-yl)-ethylamino]- methyl}-phenoxy)-propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester;

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-[3-(4-{[(R)-2-hydroxy-2- (4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}-phenoxy)- propylamino]-bicyclo[2.2.1 ]hept-7-yl ester;

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-[(3-{2-[(R)-2-hydroxy-2- (4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-ethyl}-benzyl)-methyl- amino]-bicyclo[2.2.1]hept-7-yl ester;

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-hydroxy- 2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}- benzoylamino)-ethyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester;

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-hydroxy- 2-(8-hydroxy-2-oxo-1 ,2-dihydro-quinolin-5-yl)-ethylamino]-methyl}-benzoylamino)- ethyl]-methyl-amino}-bicyclo[2.2.1 ]hept-7-yl ester;

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-hydroxy- 2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}- phenylcarbamoyO-ethyll-methyl-aminoJ-bicyclo^^.iJhept^-yl ester; Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-hydroxy-

2-(8-hydroxy-2-oxo-1 ,2-dihydro-quinolin-5-yl)-ethylamino]-methyl}-phenylcarbamoyl)- ethyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yI ester; and

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(3-{2-[(R)-2- hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-ethyl}- benzoylamino)-ethyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester; or a pharmaceutically acceptable salts thereof.

The skilled person will appreciate that each of the compounds identified above, or identified in the Examples provided herein below, taken alone or with any combination of the other identified compounds represents an independent aspect of the invention.

Compounds of the invention are β2-adrenergic binding compounds. Such compounds may be antagonists, partial agonists or full agonists. Compounds that are antagonists are useful tools, for example, for the generation of structure-activity relationships and as radioligands. Compounds that are partial or full agonists may be useful as pharmacological compounds for the treatment of diseases described hereinafter. Compounds that are antagonists may be represented by those wherein R¹⁰ is a group ArCH₂CH₂-, whilst those that are partial or full agonists may be represented by those wherein R¹⁰ is a group ArCH(OH)CH₂-.

Compounds of the invention may be useful in the treatment or prevention of diseases in which activation of muscarinic receptors are implicated, for example the present compounds are useful for treating a variety of indications, including but not limited to respiratory-tract disorders such as chronic obstructive lung disease, chronic bronchitis of all types (including dyspnoea associated therewith), asthma (allergic and non-allergic; 'wheezy-infant syndrome'), adult/acute respiratory distress syndrome (ARDS), chronic respiratory obstruction, bronchial hyperactivity, pulmonary fibrosis, pulmonary emphysema, and allergic rhinitis, exacerbation of airway hyperreactivity consequent to other drug therapy, particularly other inhaled drug therapy, pneumoconiosis (for example aluminosis, anthracosis, asbestosis, chalicosis, ptilosis, siderosis, silicosis, tabacosis and byssinosis); gastrointestinal-tract disorders such as irritable bowel syndrome, spasmodic colitis, gastroduodenal ulcers, gastrointestinal convulsions or hyperanakinesia, diverticulitis, pain accompanying spasms of gastrointestinal smooth musculature; urinary-tract disorders accompanying micturition disorders including neurogenic pollakisuria, neurogenic bladder, nocturnal enuresis, psychosomatic bladder, incontinence associated with bladder spasms or chronic cystitis, urinary urgency or pollakiuria; motion sickness; and cardiovascular disorders such as vagally induced sinus bradycardia.

For treatment of respiratory conditions, administration by inhalation will often be preferred, and in such cases administration of compounds (I) which are quaternary ammonium salts will often be preferred. In many cases, the duration of action of quaternary ammonium salts of the invention administered by inhalation is may be more than 12, or more than 24 hours for a typical dose. For treatment of gastrointestinal-tract disorders and cardiovascular disorders, administration by the parenteral route, usually the oral route, may be preferred.

Another aspect of the invention is a pharmaceutical composition comprising a compound of the invention and a pharmaceutically acceptable carrier or excipient.

Another aspect of the invention is the use of a compound of the invention or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for use in the treatment of prevention of a disease or condition in which M3 muscarinic receptor activity and β-adrenergic receptor activity are implicated.

Another aspect of the invention is a method of treatment of a disease or condition in which M3 muscarinic receptor activity and β-adrenergic receptor activity are implicated comprising administration to a subject in need thereof of an effective amount of a compound of the invention.

Another aspect of the invention is 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 muscarinic M3 receptor activity is implicated.

Another aspect of the invention is the use of a compound of formula (Ia):

wherein

R¹ is C₁-CVaIkVl or a hydrogen atom; and R² is a hydrogen atom or a group -R⁵, or a group, -Z-Y-R⁵, or a group -Z-NR⁹R¹⁰, or a group -Z-N(R⁹)C(O)R¹¹; 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;

R⁴ is selected from one of the groups of formula (a), (b), (c) or (d);

(a) (b) (C) (d) Z is a CVC₁₆-alkylene, C₂-C₁₆-aIkenylene or C₂-C₁₆-alkynylene group;

Y is a bond or oxygen atom;

R⁵ is an C_rC₆-alkyl, aryl, arylalkyl; aryl-fused-cycloalkyl, aryl-fused-heterocycloalkyl, heteroaryl, ary^d-Ce-alkyl)-, heteroary^d-Cs-alkyl)-, cycloalkyl or heterocycloalkyl group;

R⁶ is CrC₆-alkyl or a hydrogen atom; R^7a and R^7b are a C^Ce-alkyl group or halogen;

n and m are independently 0, 1 , 2 or 3;

R^8a and R^8b are independently selected from the group consisting of aryl, aryl-fused- heterocycloalkyl, heteroaryl, Ci-C₆-alkyl, cycloalkyl and hydrogen;

R^8c is -OH, Ci-C₆-alkyl, hydroxy-CrC₆-alkyl, or a hydrogen atom;

R⁹ and R¹⁰ are independently a hydrogen atom, CVC₆-alkyl, aryl, aryl-fused- heterocycloalkyl, aryl-fused-cycloalkyl, heteroaryl, aryl(C_rC₆-alkyl)-, or heteroaryl(C_r C₆-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;

R¹¹ is CrCe-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;

or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for use in the treatment of prevention of a disease or condition in which M3 muscarinic receptor activity and β-adrenergic receptor activity are implicated.

Another aspect of the invention is a method of treatment of a disease or condition in which M3 muscarinic receptor activity and β-adrenergic receptor activity is implicated comprising administration to a subject in need thereof of an effective amount of a compound of formula (Ia), as defined above.

Terminology Unless otherwise qualified in the context in which they are used herein, the following terms have the following meanings: "Acyl" means a -CO-alkyl group in which the alkyl group is as described herein. Exemplary acyl groups include -COCH₃ and -COCH(CH₃)₂.

"Acylamino" means a -NR-acyl group in which R and acyl are as described herein. Exemplary acylamino groups include -NHCOCH₃ and -N(CH₃)COCH₃. "Alkoxy" and "alkyloxy" means an -O-alkyl group in which alkyl is as described below. Exemplary alkoxy groups include methoxy (-OCH₃) and ethoxy (-OC₂H₅).

"Alkoxycarbonyl" means a -COO-alkyl group in which alkyl is as defined below. Exemplary alkoxycarbonyl groups include methoxycarbonyl and ethoxycarbonyl. "Alkyl" as a group or part of a group refers to a straight or branched chain saturated hydrocarbon group having from 1 to 12, preferably 1 to 6, carbon atoms, in the chain. Exemplary alkyl groups include methyl, ethyl, 1 -propyl and 2-propyl.

"Alkenyl" as a group or part of a group refers to a straight or branched chain hydrocarbon group having from 2 to 12, preferably 2 to 6, carbon atoms and one carbon-carbon double bond in the chain. Exemplary alkenyl groups include ethenyl, 1-propenyl, and 2-propenyl.

"Alkynyl" as a group or part of a group refers to a straight or branched chain hydrocarbon group having from 2 to 12, preferably 2 to 6, carbon atoms and one carbon-carbon triple bond in the chain. Exemplary alkenyl groups include ethynyl, 1- propynyl, and 2-propynyl.

"Alkylamino" means a -NH-alkyl group in which alkyl is as defined above. Exemplary alkylamino groups include methylamino and ethylamino.

"Alkylene means an -alkyl- group in which alkyl is as defined previously. Exemplary alkylene groups include -CH₂-, -(CH₂)₂- and -C(CH₃)HCH₂-. "Alkenylene" means an -alkenyl- group in which alkenyl is as defined previously. Exemplary alkenylene groups include -CH=CH-, -CH=CHCH₂-, and - CH₂CH=CH-.

"Alkynylene" means an -alkynyl- group in which alkynyl is as defined previously. Exemplary alkenylene groups include -CC-, -CCCH₂-, and -CH₂CC-. "Alkylsulfinyl" means a -SO-alkyl group in which alkyl is as defined above.

Exemplary alkylsulfinyl groups include methylsulfinyl and ethylsulfinyl.

"Alkylsulfonyl" means a -SO₂-alkyl group in which alkyl is as defined above. Exemplary alkylsulfonyl groups include methylsulfonyl and ethylsulfonyl.

"Alkylthio" means a -S-alkyl group in which alkyl is as defined above. Exemplary alkylthio groups include methylthio and ethylthio.

"Aminoacyl" means a -CO-NRR group in which R is as herein described. Exemplary aminoacyl groups include -CONH₂ and -CONHCH₃. "Aminoalkyl" means an alkyl-NH₂ group in which alkyl is as previously described. Exemplary aminoalkyl groups include -CH₂NH₂.

"Aminosulfonyl" means a -SO₂-NRR group in which R is as herein described. Exemplary aminosulfonyl groups include -SO₂NH₂ and -SO₂NHCH₃. "Aryl" as a group or part of a group denotes an optionally substituted monocyclic or multicyclic aromatic carbocyclic moiety of from 6 to 14 carbon atoms, preferably from 6 to 10 carbon atoms, such as phenyl or naphthyl. The aryl group may be substituted by one or more substituent groups.

"Arylalkyl" means an aryl-alkyl- group in which the aryl and alkyl moieties are as previously described. Preferred arylalkyl groups contain a C_{1 4} alkyl moiety.

Exemplary arylalkyl groups include benzyl, phenethyl and naphthlenemethyl. The aryl part thereof may be substituted by one or more substituent groups.

"Arylalkyloxy" means an aryl-alkyloxy- group in which the aryl and alkyloxy moieties are as previously described. Preferred arylalkyloxy groups contain a C alkyl moiety. Exemplary arylalkyl groups include benzyloxy. The aryl part thereof may be substituted by one or more substituent groups.

"Aryl-fused-heterocycloalkyl" means a monocyclic aryl ring, such as phenyl, fused to a heterocycloalkyl group, in which the aryl and heterocycloalkyl are as described herein. Exemplary aryl-fused-heterocycloalkyl groups include tetrahydroquinolinyl, indolinyl, benzodioxinyl, benzodioxolyl, dihydrobenzofuranyl and isoindolonyl. The aryl and heterocycloalkyl rings may each be substituted by one or more substituent groups. The aryl-fused-heterocycloalkyl group may be attached to the remainder of the compound by any available carbon or nitrogen atom.

"Aryloxy" means an -O-aryl group in which aryl is described above. Exemplary aryloxy groups include phenoxy. The aryl part thereof may be substituted by one or more substituent groups.

"Cyclic amine" is a special case of "Heterocycloalkyl" or "heterocyclic" and means an optionally substituted 3 to 8 membered monocyclic cycloalkyl ring system where one of the ring carbon atoms is replaced by nitrogen, and which may optionally contain an additional heteroatom selected from O, S or NR (where R is as described herein). Exemplary cyclic amines include pyrrolidine, piperidine, morpholine, piperazine and Λ/-methylpiperazine. The cyclic amine group may be substituted by one or more substituent groups.

"Cycloalkyl" means an optionally substituted saturated monocyclic or bicyclic ring system of from 3 to 12 carbon atoms, preferably from 3 to 8 carbon atoms, and more preferably from 3 to 6 carbon atoms. Exemplary monocyclic cycloalkyl rings include cyclopropyl, cyclopentyl, cyclohexyl and cycloheptyl. The cycloalkyl group may be substituted by one or more substituent groups.

"Dialkylamino" means a -N(alkyl)2 group in which alkyl is as defined above.

Exemplary dialkylamino groups include dimethylamino and diethylamino. "Halo" or "halogen" means fluoro, chloro, bromo, or iodo. Preferred are fluoro or chloro.

"Haloalkoxy" means an -O-alkyl group in which the alkyl is substituted by one or more halogen atoms. Exemplary haloalkyl groups include trifluoromethoxy and difluoromethoxy. "Haloalkyl" means an alkyl group which is substituted by one or more halo atoms. Exemplary haloalkyl groups include trifluoromethyl.

"Heteroaryl" as a group or part of a group denotes an optionally substituted aromatic monocyclic or multicyclic organic moiety of from 5 to 14 ring atoms, preferably from 5 to 10 ring atoms, in which one or more of the ring atoms is/are element(s) other than carbon, for example nitrogen, oxygen or sulfur. Examples of such groups include benzimidazolyl, benzoxazolyl, benzothiazolyl, benzofuranyl, benzothienyl, furyl, imidazolyl, indolyl, indolizinyl, isoxazolyl, isoquinolinyl, isothiazolyl, oxazolyl, oxadiazolyl, pyrazinyl, pyridazinyl, pyrazolyl, pyridyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, tetrazolyl, 1 ,3,4-thiadiazolyl, thiazolyl, thienyl and triazolyl groups. The heteroaryl group may be may be substituted by one or more substituent groups. The heteroaryl group may be attached to the remainder of the compound of the invention by any available carbon or nitrogen atom.

"Heteroarylalkyl" means a heteroaryl-alkyl- group in which the heteroaryl and alkyl moieties are as previously described. Preferred heteroarylalkyl groups contain a lower alkyl moiety. Exemplary heteroarylalkyl groups include pyridylmethyl. The heteroaryl part thereof may be substituted by one or more substituent groups.

"Heteroarylalkyloxy" means a heteroaryl-alkyloxy- group in which the heteroaryl and alkyloxy moieties are as previously described. Preferred heteroarylalkyloxy groups contain a lower alkyl moiety. Exemplary heteroarylalkyloxy groups include pyridylmethyloxy. The hetroaryl part thereof may be substituted by one or more substituent groups.

"Heteroaryloxy" means a heteroaryloxy- group in which the heteroaryl is as previously described. Exemplary heteroaryloxy groups include pyridyloxy. The heteroaryl part thereof may be substituted by one or more substituent groups. "Heterocycloalkyl" or "heterocyclic" means: (i) an optionally substituted cycloalkyl group of from 4 to 8 ring members which contains one or more heteroatoms selected from O, S or NR; (ii) a cycloalkyl group of from 4 to 8 ring members which contains CONR and CONRCO (examples of such groups include succinimidyl and 2-oxopyrrolidinyl). The heterocycloalkyl group may be be substituted by one or more substituents groups. The heterocycloalkyl group may be attached to the remainder of the compound by any available carbon or nitrogen atom. "Lower alkyl" as a group means unless otherwise specified, an aliphatic hydrocarbon group which may be straight or branched having 1 to 4 carbon atoms in the chain, i.e. methyl, ethyl, propyl (propyl or /so-propyl) or butyl (butyl, /so-butyl or tert- butyl).

"Sulfonyl" means a -SO₂-alkyl group in which alkyl is as described herein. Exemplary sulfonyl groups include methanesulfonyl.

"Sulfonylamino" means a -NR-sulfonyl group in which R and sulfonyl are as described herein. Exemplary sulfonylamino groups include -NHSO₂CH₃. R means alkyl, aryl, or heteroaryl as described herein. "Pharmaceutically acceptable salt" means a physiologically or toxicologically tolerable salt and includes, when appropriate, pharmaceutically acceptable base addition salts, pharmaceutically acceptable acid addition salts, and pharmaceutically acceptable quaternary ammonium salts. For example (i) where a compound of the invention contains one or more acidic groups, for example carboxy groups, pharmaceutically acceptable base addition salts that may be formed include sodium, potassium, calcium, magnesium and ammonium salts, or salts with organic amines, such as, diethylamine, Λ/-methyl-glucamine, diethanolamine or amino acids (e.g. lysine) and the like; (ii) where a compound of the invention contains a basic group, such as an amino group, pharmaceutically acceptable acid addition salts that may be formed include hydrochlorides, hydrobromides, sulfates, phosphates, acetates, citrates, lactates, tartrates, mesylates, napadisylates (naphthalene-1 ,5-disulfonates or naphthalene-1 -(sulfonic acid)-5-sulfonates), edisylates (ethane-1 ,2-disulfonates or ethane-1 -(sulfonic acid)-2-sulfonates), maleates, fumarates, succinates and the like; (iii) when R³ is not a lone pair the compound of formula (I) has a quaternary ammonium group for which the counter-ion may be, for example, chloride, bromide, sulfate, methanesulfonate, benzenesulfonate, toluenesulfonate (tosylate), napadisylate (naphthalene-1 ,5-disulfonate or naphthalene-1 -(sulfonic acid)-5- sulfonate), edisylate (ethane-1 ,2-disulfonate or ethane-1 -(sulfonic acid)-2-sulfonate), isethionate (2-hydroxyethylsulfonate), phosphate, acetate, citrate, lactate, tartrate, mesylate, maleate, malate, fumarate, xinafoate, p-acetamidobenzoate and succinate; wherein the number of quaternary ammonium species balances the counter-ion such that compound of formula (I) has no net charge. It will be understood that, as used herein, references to the compounds of the invention are meant to also include the pharmaceutically acceptable salts.

"Prodrug" refers to a compound which is convertible in vivo by metabolic means (e.g. by hydrolysis, reduction or oxidation) to a compound of the invention. For example an ester prodrug of a compound of the invention containing a hydroxy group may be convertible by hydrolysis in vivo to the parent molecule. Suitable esters of compounds of the invention containing a hydroxy group, are for example acetates, citrates, lactates, tartrates, malonates, oxalates, salicylates, propionates, succinates, fumarates, maleates, methylene-bis-β-hydroxynaphthoates, gentisates, isothionates, di-p-toluoyltartrates, methanesulfonates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates, cyclohexylsulfamates and quinates. As another example an ester prodrug of a compound of the invention containing a carboxy group may be convertible by hydrolysis in vivo to the parent molecule. Examples of ester prodrugs are those described by F. J. Leinweber, Drug Metab. Res., 1987, 18, 379. It will be understood that, as used in herein, references to the compounds of the invention are meant to also include the prodrug forms.

"Saturated" pertains to compounds and/or groups which do not have any carbon-carbon double bonds or carbon-carbon triple bonds.

"Optionally substituted" means optionally substituted with up to four susbtituents. Optional substituent groups include acyl (e.g. -COCH₃), alkoxy (e.g., -

OCHJ 3 , alkoxycarbonyl (e.g. -COOCHJ 3 , alkylamino (e.g. -NHCHJ 3 , alkylsulfinyl (e.g.

-SOCH₃), alkylsulfonyl (e.g. -SO₂CH₃), alkylthio (e.g. -SCH₃), -NH₂, aminoacyl (e.g. - CON(CH₃R aminoalkyl (e.g. -CH₂NH₂), arylalkyl (e.g. -CH₂Ph or -CH₂-CH₂-Ph), cyano, dialkylamino (e.g. -N(CH₃J₂), halo, haloalkoxy (e.g. -OCF₃ or -OCHF₂), haloalkyl (e.g. -CF 3J, alkyl (e.g. -CH 3 or -CH 2 CHJ 0 , -OH, -NO₂, aryl (optionally substituted with alkoxy, haloalkoxy, halogen, alkyl or haloalkyl), heteroaryl (optionally substituted with alkoxy, haloalkoxy, halogen, alkyl or haloalkyl), heterocycloalkyl, aminoacyl (e.g. -CONH₂, -CONHCH₃), aminosulfonyl (e.g. -SO₂NH₂, -SO₂NHCH₃), acylamino (e.g. -NHCOCH₃), sulfonylamino (e.g. -NHSO₂CH₃), heteroarylalkyl, cyclic amine (e.g. morpholine), aryloxy, heteroaryloxy, arylalkyloxy (e.g. benzyloxy) and heteroarylalkyloxy.

Alkyl, alkylene, alkenylene or alkynylene radicals may be optionally substituted. Optional substituent groups in the foregoing radicals include alkoxy (e.g., -OCHJ 3 , alkylamino (e.g. -NHCHJ 3 , alkylsulfinyl (e.g. -SOCH 3J, alkylsulfonyl (e.g. - SO 2 CH 3J, alkylthio (e.g. -SCH 3J, -NH₂, aminoalkyl (e.g. -CH c₀.NHJ -ϋ , arylalkyl (e.g. -CH Ph or -CH -CH -Ph), cyano, dialkylamino (e.g. -N(CHJJ, halo, haloalkoxy (e.g. -OCF₃ or -OCHF₂), haloalkyl (e.g. -CF₃), alkyl (e.g. -CH₃ or -CH₂CH₃), -OH, and -NO₂.

Compounds of the invention may exist in one or more geometrical, optical, enantiomeric, diastereomeric and tautomeric forms, including but not limited to cis- and trans-iorms, E- and Z-forms, R-, S- and meso-forms, keto-, and enol-forms. Unless otherwise stated a reference to a particular compound includes all such isomeric forms, including racemic and other mixtures thereof. Where appropriate such isomers can be separated from their mixtures by the application or adaptation of known methods (e.g. chromatographic techniques and recrystallisation techniques). Where appropriate such isomers may be prepared by the application of adaptation of known methods (e.g. asymmetric synthesis).

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 respiratory-tract disorders such as chronic obstructive lung disease, chronic bronchitis, asthma, chronic respiratory obstruction, pulmonary fibrosis, pulmonary emphysema, and allergic rhinitis comprising a therapeutically effective amount of a compound of the invention and one or more other therapeutic agents.

Other compounds may be combined with compounds of this invention for the prevention and treatment of inflammatory diseases of the lung. Accordingly the invention includes a combination of an agent of the invention as hereinbefore described with one or more anti-inflammatory, bronchodilator, antihistamine, decongestant or anti-tussive agents, said agents of the invention hereinbefore described and said combination agents existing in the same or different pharmaceutical compositions, administered separately or simultaneously. Preferred combinations would have two or three different pharmaceutical compositions. Suitable therapeutic agents for a combination therapy with compounds of the invention include: One or more other bronchodilators such as PDE3 inhibitors; Methyl xanthines such as theophylline; A corticosteroid, for example fluticasone propionate, ciclesonide, mometasone furoate or budesonide, or steroids described in WO02/88167, WO02/12266,

WO02/100879, WO02/00679, WO03/35668, WO03/48181 , WO03/62259,

WO03/64445, WO03/72592, WO04/39827 and WO04/66920; A non-steroidal glucocorticoid receptor agonist;

A leukotriene modulator, for example montelukast, zafirlukast or pranlukast; protease inhibitors, such as inhibitors of matrix metalloprotease for example MMP12 and TACE inhibitors such as marimastat, DPC-333, GW-3333;

Human neutrophil elastase inhibitors, such as sivelestat and those described in WO04/043942, WO05/021509, WO05/021512, WO05/026123, WO05/026124,

WO04/024700, WO04/024701 , WO04/020410, WO04/020412, WO05/080372,

WO05/082863, WO05/082864, WO03/053930;

Phosphodiesterase-4 (PDE4) inhibitors, for example roflumilast, arofylline, cilomilast,

ONO-6126 or lC-485; Phosphodiesterase-7 inhibitors;

An antitussive agent, such as codeine or dextramorphan;

Kinase inhibitors, particularly P38 MAPKinase inhibitors;

P2X7 anatgonists; iNOS inhibitors; A non-steroidal anti-inflammatory agent (NSAID), for example ibuprofen or ketoprofen;

A dopamine receptor antagonist;

TNF-α inhibitors, for example anti-TNF monoclonal antibodies, such as Remicade and CDP-870 and TNF receptor immunoglobulin molecules, such as Enbrel; A2a agonists such as those described in EP1052264 and EP1241176;

A2b antagonists such as those described in WO2002/42298;

Modulators of chemokine receptor function, for example antagonists of CCR1 , CCR2,

CCR3, CXCR2, CXCR3, CX3CR1 and CCR8, such as SB-332235, SB-656933, SB-

265610, SB-225002, MCP-1 (9-76), RS-504393, MLN-1202, INCB-3284; Compounds which modulate the action of prostanoid receptors, for example a PGD₂

(DP1 or CRTH2), or a thromboxane A₂ antagonist eg ramatroban;

Compounds which modulate Th1 or Th2 function, for example, PPAR agonists; lnterleukin 1 receptor antagonists, such as Kineret; lnterleukin 10 agonists, such as llodecakin; HMG-CoA reductase inhibitors (statins); for example rosuvastatin, mevastatin, lovastatin, simvastatin, pravastatin and fluvastatin; Mucus regulators such as INS-37217, diquafosol, sibenadet, CS-003, talnetant, DNK- 333, MSI-1956, gefitinib;

Antiinfective agents (antibiotic or antiviral), and antiallergic drugs including, but not limited to, anti-histamines.

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.

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.

The magnitude of prophylactic or therapeutic dose of a compound of the invention will, of course, vary depending upon a range 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. In general, the daily dose range for inhalation will lie within the range of from about 0.1 μg to about 10 mg per kg body weight of a human, preferably 0.1 μg to about 0.5 mg per kg, and more preferably 0.1 μg to 50μg per kg, in single or divided doses. On the other hand, it may be necessary to use dosages outside these limits in some cases. 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 1μg to 10 mg. For oral administration suitable doses are 10μg per kg to 100mg per kg, preferably 40μg per kg to 4 mg per kg.

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, and salts of quaternary ammonium compounds with pharmaceutically acceptable counter-ions.

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). Propellents 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 (C₂H₄F₂) 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, an example 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.

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). Additionally, compounds of the invention may be delivered in multi-chamber devices thus allowing for delivery of combination agents.

Methods of Synthesis

The invention further provides a process for the preparation of a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined above. In the methods described below L, L¹, L², L³, L⁴ and R¹ are as defined in formula (I) above.

Ar is as defined in R¹⁰ above. X is as defined in formula (I) above, or may be optionally suitably protected (for example such that R⁴ or R¹² does not equal hydrogen). The process comprises;

(a) when L¹ represents hydrogen, reacting a compound of formula (II)

wherein LG¹ represents a leaving group such as chloride, bromide, iodide, methanesulfonate or para-toluenesulfonate, with a compound of formula (III) or (Ilia), or a suitable salt thereof such as a hydrobromide, acetate or hydrochloride salt

wherein P¹ is hydrogen or a protective group such as tert-butyldimethyl silyl in the presence of a base such as potassium carbonate, triethylamine or diisopropylethylamine, followed by removal of the protective group (e.g. using a hydrofluoric acid-pyridine complex); or

(b)when L¹ represents hydrogen, reacting a compound of formula (IV), or a suitable salt thereof

with a compound of formula (III) or (Ilia) or a suitable salt thereof in the presence of a suitable reducing agent such as sodium cyanoborohydride, sodium triacetoxyborohydride, or hydrogen in the presence of a suitable palladium on carbon or platinum oxide catalyst; or

(c) when L¹ represents hydrogen and R¹ represents hydrogen, reacting a compound of formula (V)

wherein LG¹ represents a leaving group such as chloride, bromide, iodide, methanesulfonate or para-toluenesulfonate, P² represents a protective group (e.g. tert-butylcarbonyl) with a compound of formula (III) or (Ilia), or a suitable salt thereof

(e.g. hydrobromide, hydrochloride salt or acetate), in the presence of a base (e.g. potassium carbonate, triethylamine or diisopropylethylamine) followed by removal of the protective group (e.g. treatment with hydrochloric or trifluoroacetic acid); or

(d) when L¹ represents hydrogen and R¹ represents hydrogen, reacting a compound of formula (Vl)

wherein P² represents a protective group (e.g. tert-butylcarbonyl) with a compound of formula (III) or (Ilia), or a suitable salt thereof (e.g. hydrobromide, hydrochloride salt or acetate), in the presence of a suitable reducing agent (e.g. sodium cyanoborohydride, sodium triacetoxyborohydride, or hydrogen in the presence of a suitable palladium on carbon or platinum oxide catalyst), followed by removal of the protective group (e.g. treatment with hydrochloric or trifluoroacetic acid); or (e) when R¹ does not represent hydrogen, reacting a compound of formula (VII), or a suitable salt thereof

P³ represents hydrogen or an activating group (e.g. 3-nitrophenylsulfonyl) with a compound of formula (VIII) or (Villa), or a suitable salt thereof,

(Villa) wherein LG² represents a leaving group (e.g. chloride, bromide, iodide, methanesulfonate or para-toluenesulfonate) and P¹ is as defined in compound of formula (III) in the presence of a base (e.g. when P³ is hydrogen, potassium carbonate, triethylamine, diisopropylethylamine and, when P³ is 3-nitrophenylsulfonyl, sodium hydride or lithium di-/so-propylamide), followed by removal of the protective groups (e.g. using hydrofluoric acid-pyridine complex, thiophenol, thioacetic acid); or with a compound of formula (IX), or a suitable salt thereof,

wherein Ar is as defined in formula (I) in the presence of a base (e.g. when P³ is hydrogen, potassium carbonate, triethylamine, diisopropylethylamine and, when P³ is 3-nitrophenylsulfonyl, sodium hydride or lithium di-Zsopropylamide), followed by removal of the protective groups (e.g. trifluoroacetic acid, thiophenol, thioacetic acid); or with a compound of formula (X), or a suitable salt thereof,

wherein LG² represents a leaving group (e.g. chloride, bromide, iodide, methanesulfonate or para-toluenesulfonate) in the presence of a base (e.g. when P³ is hydrogen, potassium carbonate, triethylamine, diisopropylethylamine and, when P³ is 3-nitrophenylsulfonyl, sodium hydride or lithium di-/so-propylamide), followed by reduction of the ketone (e.g. using sodium borohydride or a borane/chiral catalyst complex), followed by removal of the protective groups (e.g. trifluoroacetic acid, thiophenol, thioacetic acid); or (f) When R¹ represents hydrogen, reacting a compound of formula (Xl)

wherein P² represents a protective group (e.g. tert-butylcarbonyl), P³ represents hydrogen or an activating group (e.g. 3-nitrophenylsulfonyl), with a compound of formula (VIII), (IX) or (X), or a suitable salt thereof, in the presence of a base (e.g. when P³ is hydrogen, potassium carbonate, triethylamine, diisopropylethylamine and when P³ is 3-nitrophenylsulfonyl, sodium hydride or lithium di-/so-propylamide), followed by removal of the protective groups (e.g. using trifluoroacetic acid, thiophenol, thioacetic acid); or

(g)when L³ and L⁴ each represents hydrogen, reacting a compound of formula (XII) or

(XIIa)

(XIIa) wherein Ar, L, L¹, and L² are as defined in formula (I), P¹ is as defined in compound of formula (III), P³ represents a protective group (e.g. tert-butylcarbonyl or 3- nitrophenylsulfonyl) with a compound of formula (XIII), or a suitable salt thereof,

in the presence of a suitable reducing agent (e.g. sodium cyanoborohydride, sodium triacetoxyborohydride, or hydrogen in the presence of a suitable palladium on carbon or platinum oxide catalyst), followed by removal of the protective groups (e.g. treatment with hydrochloric or trifluoroacetic acid thiophenol, thioacetic acid); or (h) when one or both of L³ and L⁴ represents hydrogen, reacting a compound of formula (XIV) or (XIVa)

wherein Ar, L₁ L¹, and L² are as defined in formula (I), P¹ is as defined in compound of formula (III), P³ represents a protective group (e.g. tert-butylcarbonyl or 3- nitrophenylsulfonoyl), LG³ represents a leaving group (e.g. chloride, bromide, iodide, methanesulfonate or para-toluenesulfonate), with a compound of formula (XIII) or a suitable salt thereof, in the presence of a base (e.g. potassium carbonate, triethylamine, diisopropylethylamine), followed by removal of the protective groups (e.g. trifluoroacetic acid, thiophenol, thioacetic acid); or

(i) when L¹ and L² each represents hydrogen and R¹ does not represent hydrogen, reacting a compound of formula (XV) or (XVa), or a suitable salt thereof,

wherein P¹ is as defined in formula (III) with a suitable reducing agent (e.g. borane tetrahydrofuran complex), followed by removal of the protective group (e.g. using hydrofluoric acid-pyridine complex); or,

(j)when L¹ and L² each represents hydrogen, reacting a compound of formula (XVI) or

(XVIa)

wherein P² is as defined in compound of formula (Xl) with a suitable reducing agent (e.g. borane tetrahydrofuran complex), followed by removal of the protective group (e.g. using hydrofluoric acid-pyridine complex); and optionally after (a), (b), (c), (d), (e), (f), (g), (h), (i) or (j) carrying out one or more of the following:

• converting the compound obtained to a further compound of the invention

• forming a pharmaceutically acceptable salt of the compound.

In process variants (a), (c), (e), (f) and (h), the reaction may conveniently be carried out in an organic solvent such as Λ/,Λ/-dimethylformamide, ethanol, n-butanol or dimethyl sulfoxide, at a temperature, for example, in the range from 50 to 140°C. In process variants (b), (d) and (g), the reaction may conveniently be carried out in an organic solvent such as methanol, ethanol, dichloromethane, acetic acid N- methylpyrolidinone, or Λ/,Λ/-dimethylformamide containing up to 10%w of water and acetic acid.

In process variants (i) and (j), the reaction may conveniently be carried out in an organic solvent such as tetrahydrofuran, at a temperature, for example, in the range from 0 to 80⁰C. Compounds of formula (M) may be prepared by reacting a compound of formula

(XVII), or a suitable salt thereof,

with a compound of formula (XVIII)

2 L^Mt (XVIII) wherein L² is as defined in formula (II) and Mt represents a metal such as lithium or magnesium, or aluminium or boron (e.g. methyllithium, methylmagnesium bromide, lithium aluminium hydride, sodium borohydride) in an organic solvent, for example, tetrahydrofuran or ether, at a temperature, for example in the range from 0 to 6O⁰C, followed by conversion of the resulting hydroxyl group into a suitable leaving group (e.g. chloride, bromide, iodide, methanesulfonate or para-toluenesulfonate).

Compounds of formula (IV) may be prepared by reacting a compound of formula (XVII) with a compound of formula (XVIII) in an organic solvent, for example, tetrahydrofuran or ether, at a temperature, for example in the range from 0 to 6O⁰C, followed by oxidation of the resulting hydroxyl group with a suitable oxidising agent (e.g. Swern reagent, Dess-Martin reagent or pyridiniumchlorochromate) in an organic solvent such as dichloromethane, Λ/,Λ/-dimethylformamide or dimethylsulfoxide at a temperature, for example in the range from -78 to 60⁰C. Compounds of formula (V) may be prepared by reacting a compound of formula (XIX)

wherein P², L, L³, L⁴, and X are as defined in formula (V), with a compound of formula (XVIII) in an organic solvent, for example, tetrahydrofuran or ether, at a temperature, for example in the range from 0 to 6O⁰C, followed by conversion of the resulting hydroxyl group into a suitable leaving group (e.g. chloride, bromide, iodide, methanesulfonate or para-toluenesulfonate).

Compounds of formula (Vl) may be prepared by reacting a compound of formula (XVIII) with a compound of formula (XIX), followed by oxidation of the resulting hydroxyl group with a suitable oxidating agent (e.g. Swern reagent, Dess-Martin reagent or pyridiniumchlorochromate) in an organic solvent such as dichloromethane, Λ/,Λ/-dimethylformamide or dimethylsulfoxide at a temperature, for example in the range from -78 to 6O⁰C.

Compounds of formula (VII) in which L¹ represents hydrogen and L, L², L³, L⁴, and R⁴ are as defined in formula (VII) may be prepared by (a) reacting a compound of formula (II) with sodium azide, in an organic solvent for example, tetrahydrofuran, Λ/,Λ/-dimethylformamide or dimethylsulfoxide at a temperature, for example in the range from 25 to 85⁰C, followed by reduction of the resulting azido compound using a suitable reducing agent (e.g. triphenylphosphine) in an organic solvent for example, tetrahydrofuran and water, and eventually followed by protection of the resulting amine (e.g. treatment with 3-nitrophenylsulfonyl chloride in the presence of a base such as pyridine); or,

(b) reacting a compound of formula (IV) with an amine (e.g. benzylamine, α-methyl benzylamine, 4-methoxybenzylamine or 2,4-methoxybenzylamine) followed by reduction of the resulting imine using a suitable reducing agent (e.g. sodium cyanoborohydride or sodium triacetoxyborohydride) in an organic solvent such as methanol, ethanol, dichloromethane, acetic acid, Λ/-methylpyrolidinone or N, N- dimethylformamide containing up to 10%w of water and acetic acid, followed by removal of the resulting benzyl protective group using the appropriate reagent (e.g. hydrogen and a suitable catalyst (Palladium on carbon or palladium hydroxide), 2,3- dichloro-5,6-dicyanobenzoquinone (DDQ), or ammonium cerium nitrate (CAN)) in an organic solvent, for example, ethanol, methanol, tetrahydrofuran, dichloromethane, acetonitrile, water, or a mixture thereof, at a temperature ranging from 25 to 80⁰C, and eventually followed by protection of the resulting amine (e.g. treatment with 3- nitrophenylsulfonyl chloride in the presence of a base such as pyridine);

Compounds of formula (VII) in which L, L¹, L², L³, L⁴, R¹, and X are as defined in formula (VII) may be prepared by reacting a compound of formula (XX)

wherein LG⁴ is a leaving group (e.g. hydroxyl or chloride), with reagents such as, when LG⁴ is hydroxyl, diphenylphosphonic azide, in a presence of an amine (e.g. triethylamine), in an organic solvent, for example, tert-butanol, tetrahydrofuran, dichloromethane, water, or a mixture thereof, at a temperature ranging from 25 to 100⁰C, or when LG⁴ is chloride, sodium azide, in an organic solvent, for example, ether, te/t-butanol, tetrahydrofuran, water, or a mixture thereof, at a temperature ranging from 25 to 100°C (Angewandte Chemie, 2005, 54, 5188), eventually followed by protection of the resulting amine (e.g. treatment with 3-nitrophenylsulfonyl chloride in the presence of a base such as pyridine).

Compounds of formula (III), (Ilia), (VIII), (Villa), (IX) and (X) are known in the literature or may be prepared using known techniques.

Compounds of formula (Xl) in which L¹ represents hydrogen may be prepared by (a) reacting a compound of formula (V) with sodium azide in an organic solvent, for example, tetrahydrofuran, Λ/,Λ/-dimethylformamide or dimethylsulfoxide at a temperature, for example in the range from 25 to 85⁰C, followed by reduction of the resulting azido compound using a suitable reducing agent (e.g. triphenylphosphine or hydrogen) in an organic solvent for example, tetrahydrofuran and water, eventually followed by protection of the resulting amine (e.g. treatment with 3-nitrophenylsulfonyl chloride in the presence of a base such as pyridine); or (b) reacting a compound of formula (Vl) with an amine (e.g. benzylamine, a- methyl benzylamine, 4-methoxybenzyl amine or 2,4-methoxybenzyl amine), followed by reduction of the resulting imine using a suitable reducing agent (e.g. sodium cyanoborohydride, sodium triacetoxyborohydride) in an organic solvent such as methanol, ethanol, dichloromethane, acetic acid Λ/-methylpyrolidinone, or N,N- dimethylformamide containing up to 10%w of water and acetic acid, followed by removal of the resulting benzyl protective group using the appropriate reagent (e.g. hydrogen and a suitable catalyst (Palladium on carbon or palladium hydroxide), 2,3- dichloro-5,6-dicyanobenzoquinone (DDQ), or ammonium cerium nitrate (CAN)) in an organic solvent, for example, ethanol, methanol, tetrahydrofuran, dichloromethane, acetonitrile, water, or a mixture thereof, at a temperature ranging from 25 to 80°C, eventually followed by protection of the resulting amine (e.g. treatment with 3- nitrophenylsulfonyl chloride in the presence of a base such as pyridine).

Compounds of formula (Xl) may be prepared by reacting a compound of formula (XXI)

wherein LG⁴ is a leaving group (e.g. hydroxyl or chloride), L, L¹, L², L³, L⁴, X and P² are as defined in formula (Xl), with reagents such as, when LG⁴ is hydroxyl, diphenylphosphonic azide, in a presence of an amine (e.g. triethylamine), in an organic solvent, for example, terf-butanol, tetrahydrofuran, dichloromethane, water, or a mixture thereof, at a temperature ranging from 25 to 100⁰C, or when LG⁴ is chloride, sodium azide, in an organic solvent, for example, ether, terf-butanol, tetrahydrofuran, water, or a mixture thereof, at a temperature ranging from 25 to 100⁰C (Angewandte Chemie, 2005, 54, 5188), eventually followed by protection of the resulting amine (e.g. treatment with 3-nitrophenylsulfonyl chloride in the presence of a base such as pyridine).

Compounds of formula (XII) or (XIIa) can be prepared by (a) reacting a compound of formula (XXII)

wherein P⁵ is hydrogen or a protective group (e.g. tert-butyldimethylsilyl, tetrahydropyran) and L, L¹ and L² are as defined in formula (XII), with a compound of formula (VIII), (Villa), (IX) or (X), or a suitable salt thereof, in the presence of a base (e.g. potassium carbonate, triethylamine or diisopropylethylamine when P³ is hydrogen and sodium hydride or lithium di-/so-propylamide when P³ is 3- nitrophenylsulfonyl) in an organic solvent such as Λ/,Λ/-dimethylformamide, N- methylpyrolidinone, tetrahydrofuran, ethanol, n-butanol or dimethyl sulfoxide, at a temperature, for example, in the range from 50 to 140°C. When reacting with compound of formula (X), this is followed by reduction of the ketone (e.g. using sodium borohydride or a borane/chiral catalyst complex). Appropriate selective removal of the protective group (e.g. hydrofluoric acid-pyridine complex, tetrabutylamonium fluoride, diluted hydrochloric acid or amberlyst-15 resin in methanol) and oxidation of the resulting alcohol into the corresponding aldehyde with a suitable oxidating agent (pyridinium chlorochromate, Dess-martin reagent or Swern reagent) lead to compound of formula (XII) or (XIIa); or (b) reacting a compound of formula (XXIII)

wherein P⁶ and P⁷ represent an acyclic or cyclic carbonyl protective group (e.g. dimetoxy or diethoxy acetal, 1 ,3-dioxolane or 1 ,3-dioxane) and L, L¹, L², and P³ are as defined in formula (XII), with a compound of formula (VIII), (Villa), (IX) or (X), or a suitable salt thereof, in the presence of a base (e.g. potassium carbonate, triethylamine or diisopropylethylamine when P³ is hydrogen and sodium hydride or lithium di-/so-propylamide when P³ is 3-nitrophenylsulfonyl) in an organic solvent such as Λ/,/V-dimethylformamide, Λ/-methylpyrolidinone, tetrahydrofuran, ethanol, n- butanol or dimethyl sulfoxide, at a temperature, for example, in the range from 50 to 140⁰C. When reacting with compound of formula (X), this is followed by reduction of the ketone (e.g. using sodium borohydride or a borane/chiral catalyst complex). Removal of the protective group (e.g. diluted hydrochloric acid or amberlyst-15 resin in methanol) lead to compound of formula (XII) or (XIIa); or (c) when L¹ represents hydrogen, reacting a compound of formula (XXIV)

wherein P⁵ is hydrogen or a protective group (e.g. tert-butyldimethylsilyl, tetrahydropyran) and, L and L² are as defined in formula (XII), with a compound of formula (III) or (Ilia), or a suitable salt thereof, in the presence of a suitable reducing agent (e.g. sodium cyanoborohydride, sodium triacetoxyborohydride, or hydrogen in the presence of a suitable palladium on carbon or platinum oxide catalyst) in an organic solvent such as methanol, ethanol, dichloromethane, acetic acid, N- methypyrolidinone or Λ/,Λ/-dimethylformamide containing up to 10%w of water and acetic acid, followed by appropriate selective removal of the protective group (e.g. hydrofluoric acid-pyridine complex, tetrabutylamonium fluoride, diluted hydrochloric acid or amberlyst-15 resin in methanol) and oxidation of the resulting alcohol into the corresponding aldehyde with a suitable oxidating agent (pyridinium chlorochromate, Dess-Martin reagent or Swern reagent); or (d) when R¹ represents hydrogen, reacting a compound of formula (XXV)

wherein P⁶ and P⁷ represent an acyclic or cyclic carbonyl protective group (e.g. dimethoxy or diethoxy acetal, 1 ,3-dioxolane or 1 ,3-dioxane) and, L and L² are as defined in formula (XII), with a compound of formula (III) or (Ilia), or a suitable salt thereof, in the presence of a suitable reducing agent (e.g. sodium cyanoborohydride, sodium triacetoxyborohydride, or hydrogen in the presence of a suitable palladium on carbon or platinum oxide catalyst) in an organic solvent such as methanol, ethanol, dichloromethane, acetic acid, Λ/-methypyrolidinone or Λ/,Λ/-dimethylformamide containing up to 10%w of water and acetic acid, followed by removal of the protective group (e.g. diluted hydrochloric acid or amberlyst-15 resin in methanol).

Compounds of formula (XIV) or (XIVa) can be prepared by converting compound of formula (XII) or (XIIa), or a precursor to compound of formula (XII) or (XIIa) as decribed above, chosing an appropriate sequence of reactions such as, for example, reduction of an aldehyde to an alcohol (e.g. sodium borohydride), appropriate selective removal of the protective group (e.g. hydrofluoric acid-pyridine complex, tetrabutylamonium fluoride, diluted hydrochloric acid or amberlyst-15 resin in methanol) and conversion of an alcohol into a suitable leaving group (e.g. halogen, mesylate, tosylate); or,

Compounds of formula (XV), (XVa), (XVI) and (XVIa) can be prepared by similar methods by reacting a compound of formula (XXVI)

wherein L, L³, L⁴, and X are as defined in formula (XV), P⁸ represents either R¹ as defined in compound of formula (XV) or P² as defined in compound of formula (XVI) and LG⁶ represent hydroxyl or a leaving group (e.g. chloride) with a compound of formula (III) or (Ilia), or a suitable salt thereof. When LG⁶ represents hydroxyl, the reaction is conveniently carried out in the presence of an activating reagent, for example, carbonyldiimidazole or O-(7- azabenzotriazol-1 -yl)-Λ/,Λ/,Λ/',Λ/-tetramethyluroniumhexafluorophosphate (HATU), in an organic solvent, for example, Λ/,Λ/-dimethylformamide or dichloromethane, at a temperature, for example in the range from 0 to 60⁰C, When LG⁶ represents chloride, the reaction is conveniently carried out in the presence of a base, for example, triethylamine or diisopropylethylamine in an organic solvent, for example, dichloromethane or tetrahydrofuran at a temperature, for example, in the range from 0 to 25⁰C.

Compounds of formula (XIII) wherein X = OR⁴ may be prepared according to the route outlined in Scheme 1a;

(XXXII) (XXXI)

(XIII) (XXVII) (XXVIII)

Scheme 1a

Compounds of formula (XIII) are prepared from compounds of formula (XXVII) wherein R¹ and R⁴ are as defined in formula (I) and P¹¹ is a protecting group such as t-butyloxycarbonyl,

(XXVII)

by reaction with acids such as trifluoroacetic acid or more preferably hydrochloric acid. Suitable solvents are dichloromethane or more suitably water, dioxane and tetrahydrofuran mixtures.

(XXVIII) Compounds of formula (XXVII) in which R⁴ is the group of formula (b) as defined above, may be prepared from compounds of general formula (XXVIII) wherein R¹ and P¹¹ are as defined in formula (XXVII) by reaction with a compound of general formula (XXIX):

wherein R^8a, R^8b, and R^8c are as defined in formula (b) and LG is a leaving group, for example, an O-alkyl, halogen or 1-imidazolyl group. The reaction is conducted in the presence of a strong base such as NaH or NaOMe in a solvent such as toluene, THF or dichloromethane at a range of temperatures, preferably between O⁰C and the reflux temperature of the solvent.

Compounds of formula (XXIX) can be prepared from compounds of general formula (XXX) by known methods.

Compounds of formula (XXX) are well known in the art and are readily available or can be prepared by known methods such as those described in WO01/04118.

Compounds of formula (XXVIII) can be prepared from compounds of formula (XXXI):

^p (XXXI)

wherein P¹² is a protecting group such as benzyl, 4-methoxybenyl or 3,4- dimethoxybenzyl and R¹ is as defined in formula (I), by reaction with hydrogen in the presence of a catalyst, preferably palladium hydroxide on carbon, in a polar solvent such as MeOH or EtOAc in the presence of di-tert butyl dicarbonate.

Compounds of formula (XXXI) can be prepared from compounds of formula (XXXII):

(XXXII)

by reaction with an amine of general formula (XXXIII):

R¹P¹²NH (XXXIII)

in the presence of a reducing agent such as sodium triacetoxyborohydride in a polar solvent such as THF or DCE. Compounds of formula (XXXII) can be prepared from compounds of formula

(XXXIV):

(XXXIV)

by reaction with a tin reagent, preferably Bu₃SnH and a radical initiator, preferably AIBN. The reaction can be performed in a range of solvents, preferably toluene, at a range of temperatures, preferably between room temperature and the reflux temperature of the solvent.

Compounds of general formula (XXXIV) can be prepared from compounds of general formula (XXXV):

by reaction with benzyl alcohol. The reaction is performed in the presence of a strong base such as NaH in a range of solvents, preferably THF or DMF at a range of temperatures, preferably between -78⁰C and ambient temperature.

Compounds of formula (XXXV) are known in the art and may be prepared as individual enantiomers: J. Chem. Soc. Perkin Trans I (1975) 1767-1773; Synthesis (1997), 155-166.

Additionally, the order of steps may be altered such that the steps as outlined in Scheme 1a may be performed on a suitably functionalised intermediate of formula (XXXVI)

(XXXVI)

wherein R⁴ may be a hydrogen atom or a suitable protecting group, and P⁹ and P¹⁰ represent the corresponding nitrogen substituents in compounds of formulas (II), (IV), (V), (Vl), (VII), (Xl), (XV), (XVI), (XVII), (XIX), (XX), (XXI) and (XXVI).;

Compounds of formula (XIII) wherein X is OR⁴ and R⁴ is the group of formula (a), and R¹ is as defined above, may be prepared from compounds of formula (XXVIII) wherein R¹ and P¹¹ are as defined above, by reaction with a compound of formula (XXXVII):

(XXXVII)

wherein Ar¹, R^7a, R^7b and n are as defined for formula (I). The reaction may take place in a range of non-nucleophilic organic solvents such as DMF or toluene at a range of temperatures, preferably between 0⁰C and the reflux temperature of the solvent, followed by deprotection using similar conditions as described for the preparation of compounds of formula (XIII) from compounds of formula (XXVII).

Compounds of formula (XXXVII) are well known in the art and are readily available or can be prepared by known methods. Compounds of formula (XIII) in which X is OR⁴ and R⁴ is the group of formula (c) as defined above, may be prepared from compounds of formula (XXVIII) by reaction with a compound of formula (XXXVIII):

(XXXVIII)

wherein Ar² and R^8c are as defined for formula (I) and LG⁸ is a leaving group, for example, an O-alkyl, halogen or 1-imidazolyl group. The reaction is conducted in the presence of a strong base such as NaH in a solvent such as toluene, THF or dichloromethane at a range of temperatures, preferably between 0 ⁰C and the reflux temperature of the solvent, followed by deprotection using similar conditions as described for the preparation of compounds of formula (XIII) from compounds of formula (XXVII).

(XXXIX)

Compounds of formula (XXXVIII) can be prepared from compounds of formula

(XXXIX) by known methods. Compounds of formula (XXXIX) are known compounds.

Compounds of formula (XIII) in which X is OR⁴ and R⁴ is the group of formula (d) as defined above and wherein R^8c in formula (d) is -OH, may be prepared from compounds of formula (XXVIII) by reaction with a compound of formula (XL):

The reaction is conducted in the presence of a strong base such as NaH in a solvent such as toluene, THF, preferably DMSO at a range of temperatures, preferably between 0 ⁰C and the reflux temperature of the solvent, , followed by deprotection using similar conditions as described for the preparation of compounds of formula (XIII) from compounds of formula (XXVII). Compounds of formula (XL) are well known in the art and are readily available or can be prepared by known methods.

Compounds of formula (Xlll-b) wherein X = NR R¹^ may be prepared according to the route outlined in Scheme 1b;

(XXXV) (XXXIV-b) (XXXII-b) (XXXII-C)

(XXVIII-b) (XXXI-b) (XXXII-Θ) (XXXII-d)

(XXVII-b) (Xlll-b)

Compounds of formula (Xlll-b) may be prepared from compounds of formula (XXVII-b), wherein R¹ and R⁵ and R¹² are as defined in formula (I) or R¹² is a suitable protecting group,

(XXVII-b) by reaction with acids such as trifluoroacetic acid or more preferably hydrochloric acid. Suitable solvents are dichloromethane or more suitably water, dioxane and tetrahydrofuran mixtures.

Compounds of formula (XXVII-b) wherein R⁵ is the group of formula (a), (b) or

(c) and R⁶ is H and R¹² is as defined above for formula (I) may be prepared from compounds of formula (XXVIII-b), wherein R¹ and R¹² are as defined in formula (I):

(XXVIII-b)

by reaction with a compound of formula (XXXVII), (XXIX) and (XXXVIII) respectively using methods similar to those described above.

Suitable methods for the preparation of compounds of formula (XXVIII-b), include reaction of compounds of formula (XXXI-b), wherein R¹ and R¹² are as defined in formula (I):

with hydrogen in the presence of palladium on carbon or palladium hydroxide on carbon in suitable solvents such as methanol, ethanol, acetic acid, ethyl acetate and mixtures thereof.

Compounds of formula (XXXI-b) can be prepared from compounds of formula (XXXII-e) wherein R¹ and R¹² are as defined in formula (I), or R¹² is a suitable protecting group, by reaction with di-tert-butyl dicarbonate in the presence of a suitable base such as diisopropylethylamine and suitable solvents such as THF or dichloromethane:

Compounds of formula (XXXII-e) can be prepared from compounds of formula (XXXI l-d) wherein R¹² is as defined in formula (I) or is a suitable protecting group,

by reaction with an appropriately substituted carbonyl compound of formula (XLI)

R-CHO (XLI)

in the presence of a suitable reducing agent such as a metal borohydride, especially sodium triacetoxyborohydride. The reaction can be performed in a range of solvents such as 1 ,2-dichloroethane, chloroform, dichloromethane, alcohols, optionally in the presence of an acid such as acetic acid, at a temperature from 0 ⁰C to the reflux temperature of the solvent, preferably from 0 ⁰C to ambient temperature of the solvent. Compounds of formula (XLI) are well known in the art and are readily available or can be prepared by known methods.

Compounds of Formula (XXXII-d) may be prepared from compounds of formula (XXXI l-c) wherein R¹² is as defined in formula (I) or is a suitable protecting group, by reduction of the oxime with a suitable reducing agent, such as a borohydride reagent, specifically NaBHVNiCI₂ in a suitable solvent such as methanol at a suitable temperature, such as 20° C.

^Λ(0H (XXXII-C)

Compounds of formula (XXXII-c) may be prepared from compounds of formula (XXXII-b) wherein R¹² is as defined in formula (I) or is a suitable protecting group, by treatment with hydroxylamine or a salt thereof in the presence of a suitable solvent such as methanol, optionally in the presence of a base such as sodium acetate, at a temperature between 0° C and the reflux temperature of the solvent, preferably at ambient temperature.

° (XXXII-b)

Compounds of formula (XXXII-b) may be prepared from compounds of formula (XXXIV-b) wherein R¹² is as defined above by reaction with a tin reagent, preferably Bu₃SnH and a radical initiator, preferably AIBN. The reaction can be performed in a range of solvents, preferably toluene, at a range of temperatures, preferably between ambient temperature and the reflux temperature of the solvent.

(XXXIV-b)

Compounds of formula (XXXIV-b) can be prepared from compounds of formula (XXXV) by reaction with an amine of formula (XLII) wherein R¹² is as defined in formula (I),

R¹²NHBn (XLII) The reaction may be performed in a range of solvents, preferably THF/dichloromethane at a range of temperatures, preferably between 0 and 100 ⁰C. Compounds of formula (XLII) are well known in the art and can be prepared by known methods, or are commercially available.

It will be appreciated by those skilled in the art that in the processes of the present invention certain functional groups such as hydroxyl or amino groups in the reagents may need to be protected by protecting groups. Thus, the preparation of the compounds of formula (I) may involve, at an appropriate stage, the removal of one or more protecting groups.

The protection and deprotection of functional groups is described in 'Protective Groups in Organic Chemistry', edited by J.W.F. McOmie, Plenum Press (1973) and 'Protective Groups in Organic Synthesis', 3^rd edition, T.W. Greene and P. G. M. Wuts, Wiley-lnterscience (1999).

Compounds of formula (I) can be converted into further compounds of formula (I) using standard procedures.

The compounds of formula I have activity as pharmaceuticals, in particular as dual adrenergic β2 receptor agonists and anticholinergic agents including muscarinic receptor (M1 , M2, and M3) antagonists, in particular M3 antagonists. Diseases and conditions which may be treated with the compounds of formula (I) and their pharmaceutically acceptable salts include:

1. respiratory tract: obstructive diseases of the airways including: asthma, including bronchial, allergic, intrinsic, extrinsic, exercise-induced, drug-induced (including aspirin and NSAID-induced) and dust-induced asthma, both intermittent and persistent and of all severities, and other causes of airway hyper-responsiveness; chronic obstructive pulmonary disease (COPD); bronchitis, including infectious and eosinophilic bronchitis; emphysema; bronchiectasis; cystic fibrosis; sarcoidosis; farmer's lung and related diseases; hypersensitivity pneumonitis; lung fibrosis, including cryptogenic fibrosing alveolitis, idiopathic interstitial pneumonias, fibrosis complicating anti-neoplastic therapy and chronic infection, including tuberculosis and aspergillosis and other fungal infections; complications of lung transplantation; vasculitic and thrombotic disorders of the lung vasculature, and pulmonary hypertension; antitussive activity including treatment of chronic cough associated with inflammatory and secretory conditions of the airways, and iatrogenic cough; acute and chronic rhinitis including rhinitis medicamentosa, and vasomotor rhinitis; perennial and seasonal allergic rhinitis including rhinitis nervosa (hay fever); nasal polyposis; acute viral infection including the common cold, and infection due to respiratory syncytial virus, influenza, coronavirus (including SARS) or adenovirus; or eosinophilic esophagitis;

2. bone and joints: arthritides associated with or including osteoarthritis/osteoarthrosis, both primary and secondary to, for example, congenital hip dysplasia; cervical and lumbar spondylitis, and low back and neck pain; osteoporosis; rheumatoid arthritis and Still's disease; seronegative spondyloarthropathies including ankylosing spondylitis, psoriatic arthritis, reactive arthritis and undifferentiated spondarthropathy; septic arthritis and other infection- related arthopathies and bone disorders such as tuberculosis, including Potts' disease and Poncet's syndrome; acute and chronic crystal-induced synovitis including urate gout, calcium pyrophosphate deposition disease, and calcium apatite related tendon, bursal and synovial inflammation; Behcet's disease; primary and secondary Sjogren's syndrome; systemic sclerosis and limited scleroderma; systemic lupus erythematosus, mixed connective tissue disease, and undifferentiated connective tissue disease; inflammatory myopathies including dermatomyositits and polymyositis; polymalgia rheumatica; juvenile arthritis including idiopathic inflammatory arthritides of whatever joint distribution and associated syndromes, and rheumatic fever and its systemic complications; vasculitides including giant cell arteritis, Takayasu's arteritis, Churg-Strauss syndrome, polyarteritis nodosa, microscopic polyarteritis, and vasculitides associated with viral infection, hypersensitivity reactions, cryoglobulins, and paraproteins; low back pain; Familial Mediterranean fever, Muckle-Wells syndrome, and Familial Hibernian Fever, Kikuchi disease; drug-induced arthalgias, tendonititides, and myopathies;

3. pain and connective tissue remodelling of musculoskeletal disorders due to injury [for example sports injury] or disease: arthitides (for example rheumatoid arthritis, osteoarthritis, gout or crystal arthropathy), other joint disease (such as intervertebral disc degeneration or temporomandibular joint degeneration), bone remodelling disease (such as osteoporosis, Paget's disease or osteonecrosis), polychondritits, scleroderma, mixed connective tissue disorder, spondyloarthropathies or periodontal disease (such as periodontitis); 4. skin: psoriasis, atopic dermatitis, contact dermatitis or other eczematous dermatoses, and delayed-type hypersensitivity reactions; phyto- and photodermatitis; seborrhoeic dermatitis, dermatitis herpetiformis, lichen planus, lichen sclerosus et atrophica, pyoderma gangrenosum, skin sarcoid, discoid lupus erythematosus, pemphigus, pemphigoid, epidermolysis bullosa, urticaria, angioedema, vasculitides, toxic erythemas, cutaneous eosinophilias, alopecia areata, male-pattern baldness, Sweet's syndrome, Weber-Christian syndrome, erythema multiforme; cellulitis, both infective and non-infective; panniculitis;cutaneous lymphomas, non-melanoma skin cancer and other dysplastic lesions; drug-induced disorders including fixed drug eruptions;

5. eyes: blepharitis; conjunctivitis, including perennial and vernal allergic conjunctivitis; iritis; anterior and posterior uveitis; choroiditis; autoimmune; degenerative or inflammatory disorders affecting the retina; ophthalmitis including sympathetic ophthalmitis; sarcoidosis; infections including viral , fungal, and bacterial;

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

7. abdominal: hepatitis, including autoimmune, alcoholic and viral; fibrosis and cirrhosis of the liver; cholecystitis; pancreatitis, both acute and chronic;

8. genitourinary: nephritis including interstitial and glomerulonephritis; nephrotic syndrome; cystitis including acute and chronic (interstitial) cystitis and Hunner's ulcer; acute and chronic urethritis, prostatitis, epididymitis, oophoritis and salpingitis; vulvovaginitis; Peyronie's disease; erectile dysfunction (both male and female);

9. allograft rejection: acute and chronic following, for example, transplantation of kidney, heart, liver, lung, bone marrow, skin or cornea or following blood transfusion; or chronic graft versus host disease;

10. CNS: Alzheimer's disease and other dementing disorders including CJD and nvCJD; amyloidosis; multiple sclerosis and other demyelinating syndromes; cerebral atherosclerosis and vasculitis; temporal arteritis; myasthenia gravis; acute and chronic pain (acute, intermittent or persistent, whether of central or peripheral origin) including visceral pain, headache, migraine, trigeminal neuralgia, atypical facial pain, joint and bone pain, pain arising from cancer and tumor invasion, neuropathic pain syndromes including diabetic, post-herpetic, and HIV-associated neuropathies; neurosarcoidosis; central and peripheral nervous system complications of malignant, infectious or autoimmune processes; 11. other auto-immune and allergic disorders including Hashimoto's thyroiditis, Graves' disease, Addison's disease, diabetes mellitus, idiopathic thrombocytopaenic purpura, eosinophilic fasciitis, hyper-lgE syndrome, antiphospholipid syndrome; 12. other disorders with an inflammatory or immunological component; including acquired immune deficiency syndrome (AIDS), leprosy, Sezary syndrome, and paraneoplastic syndromes;

13. cardiovascular: atherosclerosis, affecting the coronary and peripheral circulation; pericarditis; myocarditis , inflammatory and auto-immune cardiomyopathies including myocardial sarcoid; ischaemic reperfusion injuries; endocarditis, valvulitis, and aortitis including infective (for example syphilitic); vasculitides; disorders of the proximal and peripheral veins including phlebitis and thrombosis, including deep vein thrombosis and complications of varicose veins; 14. oncology: treatment of common cancers including prostate, breast, lung, ovarian, pancreatic, bowel and colon, stomach, skin and brain tumors and malignancies affecting the bone marrow (including the leukaemias) and lymphoproliferative systems, such as Hodgkin's and non-Hodgkin's lymphoma; including the prevention and treatment of metastatic disease and tumour recurrences, and paraneoplastic syndromes; and,

15. gastrointestinal tract: Coeliac disease, proctitis, eosinopilic gastro-enteritis, mastocytosis, Crohn's disease, ulcerative colitis, microscopic colitis, indeterminant colitis, irritable bowel disorder, irritable bowel syndrome, non-inflammatory diarrhea, food-related allergies which have effects remote from the gut, e.g., migraine, rhinitis and eczema.

Thus, the present invention provides a compound of formula (I) or a pharmaceutically-acceptable salt thereof as hereinbefore defined for use in therapy. In a further aspect, the present invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as hereinbefore defined in the manufacture of a medicament for use in therapy.

In the context of the present specification, the term "therapy" also includes "prophylaxis" unless there are specific indications to the contrary. The terms "therapeutic" and "therapeutically" should be construed accordingly.

Prophylaxis is expected to be particularly relevant to the treatment of persons who have suffered a previous episode of, or are otherwise considered to be at increased risk of, the disease or condition in question. Persons at risk of developing a particular disease or condition generally include those having a family history of the disease or condition, or those who have been identified by genetic testing or screening to be particularly susceptible to developing the disease or condition. The invention still further provides a method of treating, or reducing the risk of, an inflammatory disease or condition (including a reversible obstructive airways disease or condition) which comprises administering to a patient in need thereof a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof as hereinbefore defined.

In particular, the compounds of this invention may be used in the treatment of adult respiratory distress syndrome (ARDS), pulmonary emphysema, bronchitis, bronchiectasis, chronic obstructive pulmonary disease (COPD), asthma and rhinitis.

For the above-mentioned therapeutic uses the dosage administered will, of course, vary with the compound employed, the mode of administration, the treatment desired and the disorder indicated. For example, the daily dosage of the compound of the invention, if inhaled, may be in the range from 0.05 micrograms per kilogram body weight (μg/kg) to 100 micrograms per kilogram body weight (μg/kg).

Alternatively, if the compound is administered orally, then the daily dosage of the compound of the invention may be in the range from 0.01 micrograms per kilogram body weight (μg/kg) to 100 milligrams per kilogram body weight (mg/kg).

The compounds of formula (I) and pharmaceutically acceptable salts thereof may be used on their own but will generally be administered in the form of a pharmaceutical composition in which the formula (I) compound/salt (active ingredient) is in association with a pharmaceutically acceptable adjuvant, diluent or carrier. Conventional procedures for the selection and preparation of suitable pharmaceutical formulations are described in, for example, "Pharmaceuticals - The Science of Dosage Form Designs", M. E. Aulton, Churchill Livingstone, 1988.

Depending on the mode of administration, the pharmaceutical composition will preferably comprise from 0.05 to 99 %w (per cent by weight), more preferably from 0.05 to 80 %w, still more preferably from 0.10 to 70 %w, and even more preferably from 0.10 to 50 %w, of active ingredient, all percentages by weight being based on total composition.

The present invention also provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as hereinbefore defined, in association with a pharmaceutically acceptable adjuvant, diluent or carrier. The invention further provides a process for the preparation of a pharmaceutical composition of the invention which comprises mixing a compound of formula (I) or a pharmaceutically acceptable salt thereof as hereinbefore defined with a pharmaceutically acceptable adjuvant, diluent or carrier.

The pharmaceutical compositions may be administered topically (e.g. to the skin or to the lung and/or airways) in the form, e.g., of creams, solutions, suspensions, heptafluoroalkane (HFA) aerosols and dry powder formulations, for example, formulations in the inhaler device known as the Turbuhaler^®; or systemically, e.g. by oral administration in the form of tablets, capsules, syrups, powders or granules; or by parenteral administration in the form of solutions or suspensions; or by subcutaneous administration; or by rectal administration in the form of suppositories; or transdermally. Dry powder formulations and pressurized HFA aerosols of the compounds of the invention may be administered by oral or nasal inhalation. For inhalation, the compound is desirably finely divided. The finely divided compound preferably has a mass median diameter of less than 10 μm, and may be suspended in a propellant mixture with the assistance of a dispersant, such as a C₈-C₂O fatty acid or salt thereof, (for example, oleic acid), a bile salt, a phospholipid, an alkyl saccharide, a perfluorinated or polyethoxylated surfactant, or other pharmaceutically acceptable dispersant.

The compounds of the invention may also be administered by means of a dry powder inhaler. The inhaler may be a single or a multi dose inhaler, and may be a breath actuated dry powder inhaler.

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

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

For oral administration the compound of the invention may be admixed with an adjuvant or a carrier, for example, lactose, saccharose, sorbitol, mannitol; a starch, for example, potato starch, corn starch or amylopectin; a cellulose derivative; a binder, for example, gelatine or polyvinylpyrrolidone; and/or a lubricant, for example, magnesium stearate, calcium stearate, polyethylene glycol, a wax, paraffin, and the like, and then compressed into tablets. If coated tablets are required, the cores, prepared as described above, may be coated with a concentrated sugar solution which may contain, for example, gum arabic, gelatine, talcum and titanium JO dioxide. Alternatively, the tablet may be coated with a suitable polymer dissolved in a readily volatile organic solvent.

For the preparation of soft gelatine capsules, the compound of the invention may be admixed with, for example, a vegetable oil or polyethylene glycol. Hard gelatine capsules may contain granules of the compound using either the above- mentioned excipients for tablets. Also liquid or semisolid formulations of the compound of the invention may be filled into hard gelatine capsules.

Liquid preparations for oral application may be in the form of syrups or suspensions, for example, solutions containing the compound of the invention, the balance being sugar and a mixture of ethanol, water, glycerol and propylene glycol. Optionally such liquid preparations may contain colouring agents, flavouring agents, saccharine and/or carboxymethylcellulose as a thickening agent or other excipients known to those skilled in art.

The compounds of the invention may also be administered in conjunction with other compounds used for the treatment of the above conditions.

The invention therefore further relates to combination therapies wherein a compound of the invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition or formulation comprising a compound of the invention, is administered concurrently or sequentially or as a combined preparation with another therapeutic agent or agents, for the treatment of one or more of the conditions listed.

In particular, for the treatment of the inflammatory diseases such as (but not restricted to) rheumatoid arthritis, osteoarthritis, asthma, allergic rhinitis, chronic obstructive pulmonary disease (COPD), psoriasis, and inflammatory bowel disease, the compounds of the invention may be combined with the following agents: nonsteroidal anti-inflammatory agents (hereinafter NSAIDs) including non-selective cyclo- oxygenase COX-1 / COX-2 inhibitors whether applied topically or systemically (such as piroxicam, diclofenac, propionic acids such as naproxen, flurbiprofen, fenoprofen, ketoprofen and ibuprofen, fenamates such as mefenamic acid, indomethacin, sulindac, azapropazone, pyrazolones such as phenylbutazone, salicylates such as aspirin); selective COX-2 inhibitors (such as meloxicam, celecoxib, rofecoxib, valdecoxib, lumarocoxib, parecoxib and etoricoxib); cyclo-oxygenase inhibiting nitric oxide donors (CINODs); glucocorticosteroids (whether administered by topical, oral, intramuscular, intravenous, or intra-articular routes); methotrexate; leflunomide; hydroxychloroquine; d-penicillamine; auranofin or other parenteral or oral gold preparations; analgesics; diacerein; intra-articular therapies such as hyaluronic acid derivatives; and nutritional supplements such as glucosamine. The present invention still further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, together with a cytokine or agonist or antagonist of cytokine function, (including agents which act on cytokine signalling pathways such as modulators of the SOCS system) including alpha-, beta-, and gamma-interferons; insulin-like growth factor type I (IGF-1); interleukins (IL) including IL1 to 17, and interleukin antagonists or inhibitors such as anakinra; tumour necrosis factor alpha (TNF-α) inhibitors such as anti-TNF monoclonal antibodies (for example infliximab; adalimumab, and CDP-870) and TNF receptor antagonists including immunoglobulin molecules (such as etanercept) and low-molecular-weight agents such as pentoxyfylline.

In addition the invention relates to a combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, with a monoclonal antibody targeting B-Lymphocytes (such as CD20 (rituximab), MRA-alLI6R) or T-Lymphocytes (CTLA4-lg, HuMax 11-15). The present invention still further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, with a modulator of chemokine receptor function such as an antagonist of CCR1 , CCR2, CCR2A, CCR2B, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CCR10 and CCR11 (for the C-C family); CXCR1 , CXCR2, CXCR3, CXCR4 and CXCR5 (for the C-X-C family) and CX₃CRI for the C-X₃-C family.

The present invention further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, with an inhibitor of matrix metalloprotease (MMPs), i.e., the stromelysins, the collagenases, and the gelatinases, as well as aggrecanase; especially collagenase-1 (MMP-1), collagenase-2 (MMP-8), collagenase-3 (MMP-13), stromelysin-1 (MMP-3), stromelysin-2 (MMP-10), and stromelysin-3 (MMP-11) and MMP-9 and MMP-12, including agents such as doxycycline.

The present invention still further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and a leukotriene biosynthesis inhibitor, 5-lipoxygenase (5-LO) inhibitor or 5-lipoxygenase activating protein (FLAP) antagonist such as; zileuton; ABT-761 ; fenleuton; tepoxalin; Abbott- 79175; Abbott-85761 ; a N-(5-substituted)-thiophene-2-alkylsulfonamide; 2,6-di-tert- butylphenolhydrazones; a methoxytetrahydropyrans such as Zeneca ZD-2138; the compound SB-210661 ; a pyridinyl-substituted 2-cyanonaphthalene compound such as L-739,010; a 2-cyanoquinoline compound such as L-746,530; or an indole or quinoline compound such as MK-591 , MK-886, and BAY x 1005. oU

The present invention further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and a receptor antagonist for leukotrienes (LT) B4, LTC4, LTD4, and LTE4. selected from the group consisting of the phenothiazin-3-1s such as L-651 ,392; amidino compounds such as CGS-25019c; benzoxalamines such as ontazolast; benzenecarboximidamides such as BIIL 284/260; and compounds such as zafirlukast, ablukast, montelukast, pranlukast, verlukast (MK-679), RG-12525, Ro-245913, iralukast (CGP 45715A), and BAY x 7195.

The present invention still further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and a phosphodiesterase (PDE) inhibitor such as a methylxanthanine including theophylline and aminophylline; a selective PDE isoenzyme inhibitor including a PDE4 inhibitor an inhibitor of the isoform PDE4D, or an inhibitor of PDE5.

The present invention further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and a histamine type 1 receptor antagonist such as cetirizine, loratadine, desloratadine, fexofenadine, acrivastine, terfenadine, astemizole, azelastine, levocabastine, chlorpheniramine, promethazine, cyclizine, or mizolastine; applied orally, topically or parenterally.

The present invention still further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and a proton pump inhibitor (such as omeprazole) or a gastroprotective histamine type 2 receptor antagonist.

The present invention further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and an antagonist of the histamine type 4 receptor.

The present invention still further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and an alpha-1/alpha-2 adrenoceptor agonist vasoconstrictor sympathomimetic agent, such as propylhexedrine, phenylephrine, phenylpropanolamine, ephedrine, pseudoephedrine, naphazoline hydrochloride, oxymetazoline hydrochloride, tetrahydrozoline hydrochloride, xylometazoline hydrochloride, tramazoline hydrochloride or ethylnorepinephrine hydrochloride.

The present invention further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and a chromone, such as sodium cromoglycate or nedocromil sodium.

The present invention still further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, with a glucocorticoid, such as flunisolide, triamcinolone acetonide, beclomethasone dipropionate, budesonide, fluticasone propionate, ciclesonide or mometasone furoate.

The present invention further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, with an agent that modulates a nuclear hormone receptor such as PPARs.

The present invention still further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, together with an immunoglobulin (Ig) or Ig preparation or an antagonist or antibody modulating Ig function such as anti-lgE (for example omalizumab). The present invention further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and another systemic or topically-applied anti-inflammatory agent, such as thalidomide or a derivative thereof, a retinoid, dithranol or calcipotriol.

The present invention still further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and combinations of aminosalicylates and sulfapyridine such as sulfasalazine, mesalazine, balsalazide, and olsalazine; and immunomodulatory agents such as the thiopurines, and corticosteroids such as budesonide.

The present invention further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, together with an antibacterial agent such as a penicillin derivative, a tetracycline, a macrolide, a beta-lactam, a fluoroquinolone, metronidazole, an inhaled aminoglycoside; an antiviral agent including acyclovir, famciclovir, valaciclovir, ganciclovir, cidofovir, amantadine, rimantadine, ribavirin, zanamavir and oseltamavir; a protease inhibitor such as indinavir, nelfinavir, ritonavir, and saquinavir; a nucleoside reverse transcriptase inhibitor such as didanosine, lamivudine, stavudine, zalcitabine or zidovudine; or a non-nucleoside reverse transcriptase inhibitor such as nevirapine or efavirenz.

The present invention still further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and a cardiovascular agent such as a calcium channel blocker, a beta-adrenoceptor blocker, an angiotensin-converting enzyme (ACE) inhibitor, an angiotensin-2 receptor antagonist; a lipid lowering agent such as a statin or a fibrate; a modulator of blood cell morphology such as pentoxyfylline; thrombolytic, or an anticoagulant such as a platelet aggregation inhibitor. The present invention further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and a CNS agent such as an antidepressant (such as sertraline), an anti-Parkinsonian drug (such as deprenyl, L- dopa, ropinirole, pramipexole, a MAOB inhibitor such as selegine and rasagiline, a comP inhibitor such as tasmar, an A-2 inhibitor, a dopamine reuptake inhibitor, an NMDA antagonist, a nicotine agonist, a dopamine agonist or an inhibitor of neuronal nitric oxide synthase), or an anti-Alzheimer's drug such as donepezil, rivastigmine, tacrine, a COX-2 inhibitor, propentofylline or metrifonate.

The present invention still further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, and an agent for the treatment of acute or chronic pain, such as a centrally or peripherally-acting analgesic (for example an opioid or derivative thereof), carbamazepine, phenytoin, sodium valproate, amitryptiline or other anti-depressant agent-s, paracetamol, or a nonsteroidal anti-inflammatory agent.

The present invention further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, together with a parenterally or topically-applied (including inhaled) local anaesthetic agent such as lignocaine or a derivative thereof.

A compound of the present invention, or a pharmaceutically acceptable salt thereof, can also be used in combination with an anti-osteoporosis agent including a hormonal agent such as raloxifene, or a biphosphonate such as alendronate.

The present invention still further relates to the combination of a compound of the invention, or a pharmaceutically acceptable salt thereof, together with a: (i) tryptase inhibitor; (ii) platelet activating factor (PAF) antagonist; (iii) interleukin converting enzyme (ICE) inhibitor; (iv) IMPDH inhibitor; (v) adhesion molecule inhibitors including VLA-4 antagonist; (vi) cathepsin; (vii) kinase inhibitor such as an inhibitor of tyrosine kinase (such as Btk, Itk, Jak3 or MAP, for example Gefitinib or lmatinib mesylate), a serine / threonine kinase (such as an inhibitor of a MAP kinase such as p38, JNK, protein kinase A, B or C, or IKK), or a kinase involved in cell cycle regulation (such as a cylin dependent kinase); (viii) glucose-6 phosphate dehydrogenase inhibitor; (ix) kinin-B.subi . - or B.sub2. -receptor antagonist; (x) anti- gout agent, for example colchicine; (xi) xanthine oxidase inhibitor, for example allopurinol; (xii) uricosuric agent, for example probenecid, sulfinpyrazone or benzbromarone; (xiii) growth hormone secretagogue; (xiv) transforming growth factor (TGFβ); (xv) platelet-derived growth factor (PDGF); (xvi) fibroblast growth factor for example basic fibroblast growth factor (bFGF); (xvii) granulocyte macrophage colony stimulating factor (GM-CSF); (xviii) capsaicin cream; (xix) tachykinin NK.subi . or NK.sub3. receptor antagonist such as NKP-608C, SB-233412 (talnetant) or D-4418; (xx) elastase inhibitor such as UT-77 or ZD-0892; (xxi) TNF-alpha converting enzyme inhibitor (TACE); (xxii) induced nitric oxide synthase (iNOS) inhibitor; (xxiii) chemoattractant receptor-homologous molecule expressed on TH2 cells, (such as a CRTH2 antagonist); (xxiv) inhibitor of P38; (xxv) agent modulating the function of Toll-like receptors (TLR), (xxvi) agent modulating the activity of purinergic receptors such as P2X7; (xxvii) inhibitor of transcription factor activation such as NFkB, API or STATS; or (xxviii) a glucocorticoid receptor (GR-receptor) agonist.

In a further aspect the present invention provides a combination (for example for the treatment of COPD, asthma or allergic rhinitis) of a compound of formula (I) and one or more agents selected from the list comprising: o a non-steroidal glucocorticoid receptor (GR-receptor) agonist; o a PDE4 inhibitor including an inhibitor of the isoform PDE4D; o a modulator of chemokine receptor function (such as a CCR 1 receptor antagonist); o a steroid (such as budesonide); and o an inhibitor of p38 kinase function. A compound of the invention, or a pharmaceutically acceptable salt thereof, can also be used in combination with an existing therapeutic agent for the treatment of cancer, for example suitable agents include:

(i) an antiproliferative/antineoplastic drug or a combination thereof, as used in medical oncology, such as an alkylating agent (for example cis-platin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulphan or a nitrosourea); an antimetabolite (for example an antifolate such as a fluoropyrimidine like 5-fluorouracil or tegafur, raltitrexed, methotrexate, cytosine arabinoside, hydroxyurea, gemcitabine or paclitaxel); an antitumour antibiotic (for example an anthracycline such as adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin-C, dactinomycin or mithramycin); an antimitotic agent (for example a vinca alkaloid such as vincristine, vinblastine, vindesine or vinorelbine, or a taxoid such as taxol or taxotere); or a topoisomerase inhibitor (for example an epipodophyllotoxin such as etoposide, teniposide, amsacrine, topotecan or a camptothecin); (ii) a cytostatic agent such as an antioestrogen (for example tamoxifen, toremifene, raloxifene, droloxifene or iodoxyfene), an oestrogen receptor down regulator (for example fulvestrant), an antiandrogen (for example bicalutamide, flutamide, nilutamide or cyproterone acetate), a LHRH antagonist or LHRH agonist (for example goserelin, leuprorelin or buserelin), a progestogen (for example megestrol acetate), an aromatase inhibitor (for example as anastrozole, letrozole, vorazole or exemestane) or an inhibitor of 5α-reductase such as finasteride; (iii) an agent which inhibits cancer cell invasion (for example a metalloproteinase inhibitor like marimastat or an inhibitor of urokinase plasminogen activator receptor function);

(iv) an inhibitor of growth factor function, for example: a growth factor antibody (for example the anti-erbb2 antibody trastuzumab, or the anti-erbb1 antibody cetuximab [C225]), a farnesyl transferase inhibitor, a tyrosine kinase inhibitor or a serine/threonine kinase inhibitor, an inhibitor of the epidermal growth factor family (for example an EGFR family tyrosine kinase inhibitor such as £J-(3-chloro-4- fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4-amine (gefitinib, AZD1839), N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine (erlotinib, OSI-774) or 6-acrylamido-hJ-(3-chloro-4-fluorophenyl)-7-(3- morpholinopropoxy)quinazolin-4-amine (Cl 1033)), an inhibitor of the platelet-derived growth factor family, or an inhibitor of the hepatocyte growth factor family; (v) an antiangiogenic agent such as one which inhibits the effects of vascular endothelial growth factor (for example the anti-vascular endothelial cell growth factor antibody bevacizumab, a compound disclosed in WO 97/22596, WO 97/30035, WO 97/32856 or WO 98/13354), or a compound that works by another mechanism (for example linomide, an inhibitor of integrin αvβ3 function or an angiostatin); (vi) a vascular damaging agent such as combretastatin A4, or a compound disclosed in WO 99/02166, WO 00/40529, WO 00/41669, WO 01/92224, WO 02/04434 or WO 02/08213;

(vii) an agent used in antisense therapy, for example one directed to one of the targets listed above, such as ISIS 2503, an anti-ras antisense; (viii) an agent used in a gene therapy approach, for example approaches to replace aberrant genes such as aberrant p53 or aberrant BRCA1 or BRCA2, GDEPT

(gene-directed enzyme pro-drug therapy) approaches such as those using cytosine deaminase, thymidine kinase or a bacterial nitroreductase enzyme and approaches to increase patient tolerance to chemotherapy or radiotherapy such as multi-drug resistance gene therapy; or (ix) an agent used in an immunotherapeutic approach, for example ex-vivo and in-vivo approaches to increase the immunogenicity of patient tumour cells, such as transfection with cytokines such as interleukin 2, interleukin 4 or granulocyte-macrophage colony stimulating factor, approaches to decrease T-cell anergy, approaches using transfected immune cells such as cytokine-transfected dendritic cells, approaches using cytokine-transfected tumour cell lines and approaches using anti-idiotypic antibodies. OJ

The following non-limiting Examples illustrate the invention.

GENERAL EXPERIMENTAL DETAILS:

General Experimental Details:

Silica gel used for medium pressure column chromatography is 0.035 to 0.070 mm (220 to 440 mesh) (e.g. Fluka silica gel 60), and an applied pressure up to 10 psi accelerated column elution. Where thin layer chromatography (TLC) has been used, it refers to silica gel TLC using plates, typically 3 x 6 cm silica gel on aluminium foil plates with a fluorescent indicator (254 nm) (e.g. Fluka 60778). All solvents and commercial reagents were used as received. Purification by pre-packed SCX-2 cartridge refers to Isolute^® SCX-2, a strong cation exchange sorbent (Argonaut/I ST).

All compounds containing a basic centre(s) and purified by reversed-phase HPLC were obtained as the TFA salt, unless stated otherwise.

Preparative reversed-phase HPLC conditions: HPLC System 1

Purification using a C18 Gemini column (250 x 21.20 mm with 5 μm particle size, Phenomenex), eluting using linear gradients of mixtures of solvent A (water with 0.1% formic acid) and solvent B (acetonitrile with 0.1% formic acid) at a flow rate of 5- 10 mL/min with UV detection set at 230 nm.

HPLC System 2

Purification using a SunFire™ Prep C18 OBD™ column (5 micron, 19 x 50 mm column, Waters Corporation), eluting using linear gradients of mixtures of solvent A (water with 0.1% TFA) and solvent B (acetonitrile with 0.1% TFA) over 11 minutes at a flow rate of 20 mL/min with UV detection set at 220 nm.

The Liquid Chromatography Mass Spectroscopy (LC-MS) systems used: LC-MS Method 1

Micromass Platform LCT with a C18 reversed-phase column (100 x 3.0 mm i.d., Higgins Clipeus, 5 μm particle size), elution using linear gradients of mixtures of solvent A (water with 0.1% formic acid) and solvent B (acetonitrile with 0.1% formic acid) at a flow rate of 1 mL/min. Gradient:

Gradient - Timeflow ml_/min %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 ionization method: Electrospray (positive ion).

LC-MS Method 2

Micromass Platform LCT with a C18 reversed-phase column (30 x 4.6 mm i.d., Phenomenex Luna, 3 μm particle size), elution using linear gradients of mixtures of solvent A (water with 0.1% formic acid) and solvent B (acetonitrile with 0.1% formic acid) at a flow rate of 1 mL/min. Gradient:

Gradient - Timeflow 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 ZQ with a C18 reversed-phase column (30 x 4.6 mm i.d., Phenomenex Luna, 3 μm particle size), elution using linear gradients of mixtures of solvent A (water with 0.1% formic acid) and solvent B (acetonitrile with 0.1% formic acid) at a flow rate of 1 mL/min. Gradient:

Gradient - Timeflow 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 4

Waters ZMD with a C18 reversed-phase column (30 x 4.6 mm Ld., Phenomenex Luna, 3 μm particle size), elution using linear gradients of mixtures of solvent A (water with 0.1% formic acid) and solvent B (acetonitrile with 0.1% formic acid) at a flow rate of 1 mL/min. Gradient:

Gradient - Timeflow 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 (200μL/min split to MS with in-line Waters 996 DAD detection). MS ionisation method: Electrospray (positive and negative ion).

LC-MS Method 5

Waters Micromass ZQ with a C18 reversed-phase column (100 x 3.0 mm, Higgins Clipeus, 5 μm particle size), elution using linear gradients of mixtures of solvent A (water with 0.1% formic acid) and solvent B (acetonitrile with 0.1% formic acid) at a flow rate of 1 mL/min. 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 at 254 nm) MS ionisation method: Electrospray (positive ion). LC-MS Method 6

Hewlett Packard 1100 MSD using C8 reversed-phase column (50 x 2.1 mm i.d., XBridge, 3.5 μm particle size), elution using linear gradients of mixtures of solvent A (water with 5% methanol and 0.1% ammonium acetate) and solvent B (acetonitrile) at a flow rate of 1 mL/min. Gradient:

Gradient - Time flow mL/min %A %B 0.00 1.0 95 5 2.50 1.0 5 95

3.00 1.0 5 95

3.10 1.0 95 5

3.50 1.0 95 5

Detection: MS, UV (in-line UV detector).

MS ionisation method: APCI (positive ion and negative ions).

LCMS Method 7

Agilent 1100 series LC/MSD with C18 reversed-phase column (50 x 2.1 mm i.d., Waters Symmetry column, 3.5 μm particle size), eluting using linear gradients of mixtures of solvent A (water with 0.1% TFA) and solvent B (acetonitrile with 0.1% TFA) at a flow rate of 1 mL/min. Gradient:

Gradient - Time flow mL/min %A %B 0.00 1.0 95 5

1.00 1.0 95 5

9.00 1.0 5 95

10.00 1.0 5 95

Detection: Mass APCI or multimode (APCI + ESI) with UV detection set at 220 nm.

LCMS Method 8

Agilent 1100 series using a C18 reversed-phase column (30 x 4.6 mm, Waters Sunfire, 2.5 μm particle size), eluting using linear gradients of mixtures of solvent A (water with 0.1% TFA) and solvent B (acetonitrile with 0.1% TFA) at a flow rate of 2.5 mL/min. Gradient: Gradient - Time flow mL/min %A %B

0.00 2.5 95 5

0.30 2.5 95 5

2.70 2.5 5 95

2.80 2.5 5 95

2.90 2.5 95 5

Detection: MS, UV (in-line UV detector).

MS ionisation method: APCI (positive ion and negative ions).

LCMS Method 9

Finnigan AQA series using a C18 reversed-phase column (30 x 4.6 mm, Luna,

2.0 μm particle size), eluting using linear gradients of mixtures of solvent A (water with 0.1% Formic acid) and solvent B (acetonitrile with 0.1% Formic acid) at a flow rate of 2.5 mL/min. Gradient:

Gradient - Time flow mL/min %A %B

0.00 2.0 95 5

0.30 2.0 95 5

2.70 2.0 5 95

2.80 2.0 5 95

2.90 2.0 95 5

Detection: MS, ELS, UV (200 μl_/min split to ES! source). MS ionisation method: Electrospray (positive ion).

Abbreviations used in the experimental section:

AIBN = (2,2'-azobis(2-methylproprionitrile)

(R)-(+)-CBS = (R)-(+)-methyl-CBS-oxazaborolidine Boc anhydride = di-tert-butyl dicarbonate

CDI = 1 ,1'-carbonyldiimidazole

DCE = 1 ,2-dichloroethane

DCM = dichloromethane

DIPEA = diisopropylethylamine DMF = Λ/,Λ/-dimethylformamide

DMSO = dimethylsulfoxide e.e. = enantiomeric excess EtOAc = ethyl acetate

EtOH = ethanol equiv. = equivalents h = hour(s) HCI = hydrochloric acid

HATU = 0-(7-azabenzotriazol-1 -yl)-Λ/,Λ/,/V'/V -tetramethyluroniumhexafluoro- phosphate

HPLC = high performance liquid chromatography

MeCN = acetonitrile MeOH = methanol min = minutes

NaHCO₃ = sodium hydrogen carbonate

NaOH = sodium hydroxide

Na₂SO₄ = sodium sulphate NH₄CI = ammonium chloride

NMP = Λ/-methyl-pyrrolidone pTSA = para-toluenesulphonic acid quant. = quantitative

RT = room temperature Rf = retention factor (TLC)

Rt = retention time (LCMS or HPLC)

SCX-2 = strong cation exchange resin

TBDMS = tert-butyldimethylsilyl

TFA = trifluoroacetic acid THF = tetrahydrofuran

TLC = thin layer chromatography

Intermediates:

Intermediate 1

(1 R,4R,5R,7S)-7-Benzyloxy-5-bromo-bicyclo[2.2.1 ]heptan-2-one

To a suspension of sodium hydride (60% dispersion in oil; 4.53 g, 113.21 mmol) in dry THF (120 mL) under N₂ was added benzyl alcohol (11.18 g, 108.07 mmol). The reaction was warmed to 35 ⁰C for 30 min then cooled to -30 ⁰C. A solution of (1R,2S,3S,5R)-2,3-dibromo-bicyclo[3.2.0]heptan-6-one (13.79 g, 51.46 mmol) in dry THF (60 mL) was added and the reaction mixture allowed to attain ambient temperature over 2 h. The reaction was quenched with 10% aqueous citric acid solution and the organic extracts washed with water and brine, dried over MgSO₄ and concentrated in vacuo. The crude product was purified by chromatography over silica gel on a Teledyne ISCO CombiFlash using 0-15% diethyl ether/cyclohexane as eluent to give the title compound as a colourless syrup.

Yield: 5.19 g (34%).

¹H NMR (400 MHz, CDCI₃): δ 7.4 - 7.3 (m, 5H), 5.3 (s, 1 H)₁ 4.75 - 4.65 (m,

1 H), 4.55 - 4.45 (m, 2H), 4.05 (s, 1 H), 2.90 - 2.75 (m, 2H), 2.65 - 2.60 (m, 1 H),

2.30 - 2.20 (m, 1 H), 1.85 - 1.75 (m, 1 H).

Intermediate 2 (1R,4R,7R)-7-Benzyloxy-bicyclo[2.2.1]heptan-2-one

To a mixture of (1 R,4R,5R,7S)-7-benzyloxy-5-bromo-bicyclo[2.2.1]heptan-2- one (5.19 g, 17.58 mmol) and AIBN (289 mg) in 210 mL dry degassed toluene was added drop-wise tributyltin hydride (6.15 mL, 22.86 mmol) and the solution was heated to 80 ⁰C for 6.5 h. The reaction mixture was concentrated under reduced pressure and purified by chromatography eluting with 0-25% diethyl ether/cyclohexane to give the title compound as a colourless gum. Yield: 2.76 g (73%).

¹H NMR (400 MHz, CDCI₃): δ 7.40 - 7.20 (m, 5H), 4.50 (m, 2H), 3.90 (m, 1 H), 2.70 - 2.65 (s, 1 H), 2.65 - 2.60 (s, 1 H), 2.20 - 1.95 (m, 4H)₁ 1.40 - 1.55 (m, 2H).

Intermediate 3 Benzyl-((1S,2S,4R,7R)-7-benzyloxy-bicyclo[2.2.1]hept-2-yl)-methyl-amine

A solution of (1 R,4R,7R)-7-benzyloxy-bicyclo[2.2.1]heptan-2-one (387 mg, 1.79 mmol) in dry DCE was stirred with activated 3A sieves (1.0 g).

Benzylmethylamine (434 mg, 3.58 mmol), sodium triacetoxyborohydride (570 mg, 2.69 mmol) and acetic acid (102 μL, 1.79 mmol) were added and the reaction stirred at RT for 18 hours. The reaction was quenched with 6 ml_ saturated aqueous NaHCO₃ solution, extracted with DCM and the combined organic phases dried over anhydrous MgSO₄ and concentrated in vacuo. Purification by chromatography using 0-1 % MeOH/DCM as eluent gave the title compound as an oil. Yield: 446 mg (73%).

¹H NMR (400 MHz, CDCI₃): 57.39 - 7.20 (m, 10H), 4.50 (s, 2H), 3.78 (s, 1 H), 3.45 - 3.29 (m, 2H), 2.46 - 2.35 (m, 2H), 2.25 - 2.20 (m, 1 H), 2.08 - 1.87 (m, 5H), 1.85 - 1.75 (m, 1 H), 1.72 - 1.60 (m, 1 H), 1.43 - 1.35 (m, 1 H), 1.12 - 1.05

(m, 1 H).

Intermediate 4

((1S,2S,4R,7R)-7-Hydroxy-bicyclo[2.2.1]hept-2-yl)-methyl-carbamic acid-te/t- butyl ester

A solution of benzyl-((1 S,2S,4R,7R)-7-benzyloxy-bicyclo[2.2.1]hept-2-yl)- methyl-amine (446 mg, 1.38 mmol) and di-te/t-butyldicarbonate (333 mg, 1.52 mmol) in dry EtOH (10 ml_) was degassed using Argon and 20 wt% palladium hydroxide on carbon (45 mg) was added. The reaction was stirred at RT under an atmosphere of hydrogen (hydrogen balloon) for 20 hours. The reaction mixture was purged with nitrogen and then filtered through HyFIo and the filtrate concentrated to give the title compound, which was used without further purification.

Yield: 335 mg (quant.).

¹H NMR (400 MHz, CDCI₃): δ 4.10 - 4.03 (m, 2H), 2.88 (s, 3H), 2.38 - 2.33 (m, 1 H), 2.09 - 2.05 (m, 1 H), 1.98 - 1.85 (m, 2H), 1.80 - 1.70 (m, 1 H), 1.65 - 1.30

(m, 13H).

Intermediate 5

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-(fert-butoxycarbonyl- methyl-amino)-bicyclo[2.2.1]hept-7-yl ester

A solution of ((1 S,2S,4R,7R)-7-hydroxy-bicyclo[2.2.1]hept-2-yl)-methyl- carbamic acid-tert-butyl ester (480 mg, 2.0 mmol) was prepared in toluene (50 ml_) in a round bottom flask fitted with short-path distillation apparatus. Hydroxy-di-thiophen- 2-yl-acetic acid ethyl ester (590 mg, 2.2 mmol) and sodium methoxide (108 mg, 2.0 mmol) were added and the mixture gradually heated until distillation of methanol (bpt 70 ⁰C, external temp. 130 ⁰C) was observed. After this fraction distilled, the mixture was heated further until toluene began to distil (external temp. 140 ⁰C). After approximately 10 ml_ toluene had been distilled the mixture was allowed to cool to room temperature. The mixture was poured into aqueous NH₄CI (50 ml_) and extracted with EtOAc (2 x 100 ml_). The combined organics were dried over anhydrous Na₂SO₄, filtered and evaporated. Purification over silica gel on a Teledyne ISCO companion eluting with a gradient of 0-30% EtOAc/cyclohexane gave the title compound as a pale yellow solid. Yield: 680 mg (73%).

¹H NMR (400 MHz, CDCI₃) δ 7.29 - 7.26 (m, 2H), 7.17 - 7.14 (m, 2H)₁ 6.99 - 6.95 (m, 2H), 4.91 (s, 1 H), 4.74 (s, 1 H), 4.12 - 4.06 (m, 1 H), 2.85 (s, 3H), 2.60 (m, 1 H), 2.31 (m, 1 H), 2.02 - 1.93 (m, 1 H), 1.55 - 1.44 (m, 3H), 1.43 (s, 9H),

1.35 - 1.25 (m, 2H). Intermediate 6

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-methylamino- bicyclo[2.2.1]hept-7-yl ester

A solution of hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-(terf- butoxycarbonyl-methyl-aminoJ-bicyclop^.ilhept-T-yl ester (1.4 g, 3.0 mmol) was formed in THF (10 mL) with 1 ,4-dioxane (2 ml_). HCI (2 M, 10 ml_) was added and the mixture was stirred vigorously and warmed to 50 ⁰C for 2 h. The mixture was allowed to cool to RT, then neutralised carefully using saturated aqueous NaHCO₃ (50 mL). The mixture was extracted with EtOAc and the combined organics were dried over Na₂SO₄, filtered and evaporated. Purification over silica gel on a Teledyne ISCO companion eluting with a gradient of 0-10% MeOH/DCM gave the title compound as a cream solid.

Yield: 800 mg (73%).

¹H NMR (400 MHz, CDCI₃): δ 7.29-7.27 (m. 2H), 7.18 - 7.16 (m, 2H), 6.99 - 6.96 (m, 2H) 4.87 (s, 1 H), 3.04 - 2.99 (m, 1 H), 2.45 (m, 1 H), 2.30 (s, 3H), 2.24 (m, 1 H), 2.05 - 1.96 (m, 1 H), 1.71 - 1.64 (m, 1 H), 1.61 - 1.52 (m, 1 H), 1.41 - 1.32 (m, 1 H), 1.30 - 1.23 (m, 1 H), 0.82 (dd, 1 H).

LC-MS (Method 5): Rt 5.91 min, m/z 364 [M+H]⁺.

Intermediate 7

[2-(4-Hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethyl]-(4-nitro-benzyl)- carbamic acid terf-butyl ester

7-((R)-2-amino-1-hydroxy-ethyl)-4-hydroxy-3H-benzothiazol-2-one hydrochloride (5 g, 20.33 mmol) (prepared according to WO2007/027133), 4- nitrobenzaldehyde (3.06 g, 20.33 mmol) and acetic acid (1.20 ml_) were stirred in a mixture of NMP (40 mL) and MeOH (10 mL) for 1 h. The mixture was cooled in an ice-bath and sodium triacetoxyborohydride (6.44 g, 30.52 mmol) was added portion- wise over 10 min. The resulting mixture was then stirred at 0 ⁰C for 30 min, then allowed to warm to RT and stirred overnight. The reaction mixture was poured into water (400 mL) and extracted with EtOAc (3 x 100 mL). The combined organic layers were washed with water, brine, dried (Na₂SO₄), filtered and concentrated in vacuo. The crude material was re-dissolved in a mixture of DCM (100 mL) and DMF (20 mL) and di-te/t-butyl dicarbonate (4.87 g, 22.33 mmol) was added. The resulting mixture was stirred overnight. Further di-te/t-butyl dicarbonate (2 g) was added and the mixture was stirred overnight. The reaction was then concentrated and poured into a mixture of water (200 mL) and EtOAc (3 x 200 mL). The combined organic extracts were washed with concentrated aqueous ammonia solution (100 mL), water, brine, dried (Na₂SO₄), filtered and concentrated in vacuo. The residue was purified by column chromatography eluting with 0-50% EtOAc/i-hexane to afford the compound as a yellow foam. Yield: 5.4 g (82%).

¹H NMR (400 MHz, d₆-DMSO, 90 ⁰C): δ 8.16 - 8.1 1 (m, 2H), 7.44 (d, J = 9.0 Hz, 2H), 6.75 (d, J = 8.5 Hz, 1H), 6.67 (d, J = 8.2 Hz, 1 H), 4.45 (s, 2H), 3.41 (t, J = 6.9 Hz, 2H), 2.68 (t, J = 7.2 Hz, 2H), 1.36 (s, 9H) + 2 exchangeable hydrogens not observed. LC-MS (Method 3): Rt 3.55 min, m/z 444 [M-H].

Intermediate 8

(4-Amino-benzyl)-[2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethyl]- carbamic acid terf-butyl ester

/o

[2-(4-Hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethyl]-(4-nitro-benzyl)- carbamic acid ferf-butyl ester (2.2 g, 4.94 mmol) and 10% palladium on carbon (0.526 g) in THF (100 ml_) were hydrogenated under 4 bar of hydrogen atmosphere at RT for 18 h. The reaction mixture was filtered through Celite and the filter pad was washed with THF (2 x 100 ml_). The combined filtrate and washings were evaporated in vacuo. The residue was re-dissolved in THF (100 ml_) and further 10% palladium on carbon (0.526 g) was added and the resulting mixture hydrogenated under 4 bar of hydrogen atmosphere at RT for 18 h. The reaction mixture was filtered through Celite and the filter pad was washed with THF (2 x 100 ml_). The combined filtrate and washings were concentrated in vacuo and azeotroped with toluene (100 ml_) to give the compound as a yellow solid.

Yield: 2.0 g (97%).

¹H NMR (400 MHz, d₆-DMSO): δ 6.89 (d, J = 8.5 Hz, 2H), 6.72-6.65 (m, 2H),

6.56 - 6.51 (m, 2H), 4.70 (s, 2H), 4.14 (s, 2H), 3.25 (t, J = 7.3 Hz, 2H), 2.59 (t, J = 7.4 Hz, 2H), 1.38 (s, 9H) + 2 exchangeable hydrogens not observed.

LC-MS (Method 3): Rt 2.72 min, m/z 414 [M-H]^".

Intermediate 9

[4-(2-Bromo-acetylamino)-benzyl]-[2-(4-hydroxy-2-oxo-2,3-dihydro- benzothiazol-7-yl)-ethyl]-carbamic acid fert-butyl ester

A cooled (0 ⁰C) solution of (4-amino-benzyl)-[2-(4-hydroxy-2-oxo-2,3-dihydro- benzothiazol-7-yl)-ethyl]-carbamic acid terf-butyl ester (0.61 g, 1.47 mmol) in DMF (20 ml.) was treated with solid NaHCO₃ (140 mg, 1.61 mmol) and allowed to stir for 15 minutes prior to the addition of bromo-acetyl bromide (0.26 ml_, 2.94 mmol at 0 °C) over 10 minutes. The reaction mixture was allowed to warm to RT overnight and was then partitioned between water and EtOAc. The combined organic extracts were concentrated in vacuo. The crude product was triturated with saturated aqueous NaHCO₃ with the aid of prolonged sonication and then collected by filtration. The solid residue was further purified using a Teledyne ISCO CombiFlash Companion, loading in EtOAc/MeOH and eluting with 0-5% MeOH/DCM to afford the title product. LC-MS (Method 2): Rt 3.45 min, m/z 536, 538 [M+H]⁺.

Intermediate 10

(4-Acryloylamino-benzyl)-[2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol- 7-yl)-ethyl]-carbamic acid fert-butyl ester

The title compound was prepared from (4-amino-benzyl)-[2-(4-hydroxy-2-oxo- 2,3-dihydro-benzothiazol-7-yl)-ethyl]-carbamic acid tert-butyl ester and acryloyl chloride by a similar method to that employed for intermediate 9.

¹H NMR (400 MHz, d₆-DMSO): δ 10.13 (s, 1 H), 7.63 (d, J = 8.5 Hz, 2H), 7.21- 7.15 (m, 2H), 6.80 - 6.66 (m, 2H), 6.43 (dd, J = 17.0, 10.1 Hz, 1 H), 6.25 (dd, J

= 17.0, 1.9 Hz, 1 H), 5.74 (dd, J = 10.0, 2.1 Hz, 1H), 4.36 - 4.17 (m, 2H), 3.41- 3.21 (m, 2H), 2.66-2.56 (m, 2H), 1.42-1.24 (m, 9H) + 2 exchangeable hydrogens not observed.

Intermediate 11

[2-(4-Amino-phenyl)-ethyl]-[2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)- ethyl]-carbamic acid fert-butyl ester

The title compound was prepared from 7-(2-amino-ethyl)-4-hydroxy-3H- benzothiazol-2-one hydrochloride and (4-nitro-phenyl)-acetaldehyde by similar methods to those employed for intermediates 7-9, respectively.

LC-MS (Method 2): Rt 3.46 min, m/z 550, 552 [M-H]^".

Intermediate 12 4-[2-(fert-Butyl-dimethyl-silanyloxy)-ethyl]-yV-(2,2-dimethoxy-ethyl)-benzamide

To a stirred solution of 4-(2-(tert-butyldimethylsilyloxy)ethyl)benzoic acid (0.70 g, 2.50 mmol) (Nucleic Acids Research 1997, 25(12), 2352-2358) in dry DMF (10 ml_) was added di(1 /-/-imidazol-1-yl)methanone (0.52 g, 3.21 mmol). After 1 h 2,2- dimethoxyethanamine (0.69 g, 6.56 mmol) was added and the solution allowed to stir at RT for a further 30 min. The solution was diluted with ethyl acetate, washed with water (x 3), brine, and then evaporated in vacuo. Purification was undertaken by silica gel chromatography, eluting with EtOAc/i-hexane, (1 :2) to afford the title compound.

Yield: 0.85 g (93%). 1H NMR (400 MHz, CDCI₃): δ 7.72 (d, J = 8.2 Hz, 2H), 7.30 (d, J = 8.2 Hz,

2H), 6.35 - 6.27 (m, 1 H), 4.52 (t, J = 5.3 Hz, 1 H), 3.84 (t, J = 6.8 Hz, 2H), 3.63 (t, J = 5.6 Hz, 2H), 3.47 (s, 6H), 2.88 (t, J = 6.8 Hz, 2H), 0.89 (s, 9H), 0.00 (s, 6H).

Intermediate 13

Λ/-(2,2-Dimethoxy-ethyl)-4-(2-hydroxy-ethyl)-benzamide

4-[2-(tert-Butyl-dimethyl-silanyloxy)-ethyl]-/V-(2,2-dimethoxy-ethyl)-benzamide

(0.83 g, 2.26 mmol) was dissolved in tetrabutylammonium fluoride (10 ml. of a 1 M solution in THF). After 0.5 h, the solution was diluted with EtOAc and washed with a small amount of water and brine. The aqueous washings were combined, extracted with EtOAc and the combined organic solutions were evaporated in vacuo.

Purification was carried out by silica gel chromatography eluting with DCM/MeOH

(10:1) to afford the title product as a solid.

Yield: 0.55 g (96%). 1H NMR (400 MHz, CDCI₃): δ 7.72 (d, J = 38.1 Hz, 2H), 7.32 (d, J = 38.1 Hz,

2H), 6.34 - 6.25 (m, 1 H), 4.49 (t, J = 19.9 Hz, 1 H), 3.94 - 3.84 (m, 2H), 3.60 (t, J = 19.9 Hz, 2H), 3.44 (s, 6H), 2.92 (t, J = 17.4 Hz, 4H).

Intermediate 14 /V-(2,2-Dimethoxy-ethyl)-4-(2-oxo-ethyl)-benzamide

Dess-Martin periodinane (1.22 g, 2.88 mmol) was added to Λ/-(2,2-dimethoxy- ethyl)-4-(2-hydroxy-ethyl)-benzamide (0.55 g, 2.17 mmol) in DCM (15 mL). After 1 h saturated aqueous NaHCO₃ (25 mL), sodium thiosulphate (25 mL) and EtOAc (80 mL) were added. The mixture was shaken vigorously for 1 min and then separated. The organic layer was washed with saturated aqueous NaHCO₃ (25 ml) and brine, dried (Na₂SO₄), filtered and evaporated in vacuo (bath temperature <30 ⁰C) to give the crude intermediate aldehyde 0.48 g (88%). A solution of 7-(2-aminoethyl)-4- hydroxybenzo[d]thiazol-2(3H)-one hydrochloride (0.54 g, 2.19 mmol) and acetic acid (0.12 mL) in NMP (15 mL) and water (5 mL) was added to the crude Λ/-(2,2- dimethoxyethyl)-4-(2-oxoethyl)benzamide (0.48 g, 1.91 mmol). Sodium triacetoxyborohydride (0.70 g) was then added and the mixture stirred at RT for 30 min. The reaction mixture was partitioned between EtOAc and saturated aqueous NaHCO₃ (50 mL). The aqueous layer was further extracted with EtOAc (x4) and the combined organic extracts were washed with brine, dried (Na₂SO₄), filtered and concentrated in vacuo to give the crude amine intermediate compound in NMP (~5 mL). Boc anhydride (0.57 ml, 2.5 mmol) was added to the crude Λ/-(2,2-dimethoxy- ethyl)-4-{2-[2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-ethyl}- benzamide in NMP (~5 mL) and DCM (10 mL). After 1 h the reaction mixture was diluted with EtOAc, washed with water, dried (Na₂SO₄), filtered, and concentrated in vacuo. Purification was carried out by silica gel chromatography eluting with EtOAc/iso-hexanes (2:1) to afford the title product as beige solid Yield: 0.38 g (32%, 3 steps).

¹H NMR (400 MHz, d₆-DMSO, 90 ⁰C): δ 8.01 - 8.1 1 (m, 1 H), 7.75 (d, J = 8.2 Hz, 2H), 7.22 (d, J = 8.2 Hz, 2H), 6.76 (d, J = 8.4 Hz, 1 H), 6.68 (d, J = 8.4 Hz,

1 H), 4.51 (t, J = 5.5 Hz, 1 H), 3.36 (t, J = 5.6 Hz, 2H), 3.27 - 3.33 (m, 4H), 3.30 (s, 6H), 2.77 (t, J = 7.6 Hz, 2H), 2.64 (t, J = 7.3 Hz, 2H), 1.34 (s, 9H).

Intermediate 15 [2-(4-Hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethyl]-{2-[4-(2-oxo- ethylcarbamoyl)-phenyl]-ethyl}-carbamic acid terf-butyl ester

{2-[4-(2,2-Dimethoxy-ethylcarbamoyl)-phenyl]-ethyl}-[2-(4-hydroxy-2-oxo-2,3- dihydro-benzothiazol-7-yl)-ethyl]-carbamic acid terf-butyl ester (2.1 g, 3.8 mmol) in acetic acid (20 ml_) and water (20 ml_) was heated at 50 ^°C for 6 h. After cooling to RT the reaction mixture was diluted with EtOAc (300 mL), washed with water (4 x 100 ml_), dried (Na₂SO₄), filtered, and concentrated in vacuo to afford the title compound, which was used directly without characterization.

Intermediate 16 /V-(2,2-Dimethoxy-ethyl)-4-hydroxymethyl-benzamide

HATU (8.30 g, 21.83 mmol) was added portion-wise over 10 min to a cooled 0 ⁰C stirred solution of 4-(hydroxymethyl)benzoic acid (2.60 g, 17.09 mmol), 2,2- dimethoxyethanamine (3.00 g, 28.53 mmol) and triethylamine (8.00 g, 79.06 mmol) in DMF (40 mL). After 1 h the reaction mixture was quenched with water (200 mL) and extracted with EtOAc (6 x 350 mL). The organic extracts were combined, dried ol

(MgSO₄), filtered and evaporated in vacuo. The crude product was purified by flash / silica chromatography, eluting with EtOAc//-hexanes (3:1) to afford the title compound.

¹H NMR (300 MHz, CDCI₃): δ 7.77 (d, J = 8.0 Hz, 2H), 7.43 (d, J = 8.0 Hz, 2H), 6.4 - 6.27 (m, 1 H), 4.76 (s, 2H), 4.50 (t, J = 5.4 Hz, 1 H), 3.61 (t, J = 5.4

Hz, 2H), 3.44 (s, 6H).

LC-MS (Method 2): Rt 2.19 min, m/z 240 [M+H]⁺.

Intermediate 17 tø-(2,2-Dimethoxy-ethyl)-4-formyl-benzamide

Manganese(IV) oxide (3.38 mL, 195.54 mmol) was added to stirred solution of Λ/-(2,2-dimethoxy-ethyl)-4-hydroxymethyl-benzamide (4.00 g, 16.72 mmol) in DCM (200 mL) at RT. After 2 h the mixture was filtered through a pad of Celite, washed with DCM, and the solution evaporated in vacuo to afford the crude aldehyde, which was used directly without characterization.

Yield: 3.9 g (98%).

Intermediate 18

[2-(4-Hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethyl]-[4-(2-oxo-et hylcarbamoyl)-benzyl]-carbamic acid tert-butyl ester

The title compound was prepared from Λ/-(2,2-dimethoxy-ethyl)-4-formyl- benzamide and 7-(2-amino-ethyl)-4-hydroxy-3/-/-benzothiazol-2-one hydrochloride by similar methods to those employed for intermediates 14 (reductive amination and Boc-protection stages) and 15, and was used directly without characterization.

Intermediate 19 (S-Acetoxymethyl-phenylJ-acetic acid methyl ester

A solution of methyl 2-(3-(bromomethyl)phenyl)acetate (1.0 g, 4.11 mmol) and sodium acetate (337 mg, 4.1 1 mmol) in DMA (15 ml_) were heated under microwave irradiation at 120 °C for 40 min. The procedure was repeated a further 5 times (6 g in total of 2-(3-(bromomethyl)phenyl)acetate was reacted). The combined reaction mixtures were diluted with EtOAc (70 ml_) and washed with water. The organic layers were dried (Na₂SO₄), filtered and concentrated in vacuo to afford the title compound as a gum.

Yield: 4.77g (93%).

¹H NMR (400 MHz, d₆-DMSO): δ 7.35 - 7.29 (m, 1 H), 7.28 - 7.21 (m, 3H),

5.05 (s, 2H), 3.69 (s, 2H), 3.61 (s, 3H), 2.08 - 2.04 (m, 3H).

Intermediate 20

(3-Hydroxymethyl-phenyl)-acetic acid

Lithium hydroxide monohydrate (2.66 mL, 95.62 mmol) was added to

(3-acetoxymethyl-phenyl)-acetic acid methyl ester (4.25 g, 19.12 mmol) in THF (40 mL) and water (40 mL) at RT under a nitrogen atmosphere. The resulting suspension was stirred at RT for 2 h. The reaction mixture was cooled on ice and 2.5 M HCI (50 mL) was added. THF removed in vacuo, the remaining aqueous layer was extracted with EtOAc, and the combined organics were dried (MgSO₄), filtered and solvent removed in vacuo to afford title compound as a tan coloured gum. Yield: 3.30 g (quant.) ¹H NMR (400 MHz, CJ₆-DMSO): δ 7.26 (t, J = 7.6 Hz, 1 H), 7.2 - 7.16 (m, 2H), 7.11 <d, J = 7.2 Hz, 1 H), 4.47 (s, 2H), 3.54 (s, 2H).

Intermediate 21 [3-(terf-Butyl-dimethyl-silanyloxymethyl)-phenyl]-acetic acid

Imidazole (3.26 g, 47.84 mmol) and TBDMS-CI (3.46 g, 22.96 mmol) were added to a solution in DMF (32 ml.) of (3-hydroxymethyl-phenyl)-acetic acid (3.18 g, 19.14 mmol) at RT. The resulting mixture was stirred at 50 °C for 2 h. The reaction mixture was cooled to RT, stirred overnight, and then heated to 50 ⁰C for 8 h. A further quantity of TBDMS-CI (0.12 equiv.) and imidazole (0.25 equiv.) were added and heating was continued overnight.. The reaction mixture was diluted with EtOAc, washed with water. The combined organic layers were dried (Na₂SO₄), filtered and solvent removed in vacuo. The taken up in EtOAc (100 mL) and washed a further 3 times with water, dried (MgSO₄), filtered and concentrated in vacuo to afford title compound as an orange gum.

Yield: 4 g (75%) 1H NMR (400 MHz, d₆-DMSO): δ 7.34 - 7.28 (m, 1 H), 7.26 - 7.20 (m, 2H),

7.19 - 7.14 (m, 1 H), 4.75 - 4.70 (m, 2H), 3.60 - 3.54 (m, 2H), 0.97 - 0.91 (m, 9H), 0.13 - 0.08 (m, 6H).

Intermediate 22 2-[3-(fert-Butyl-dimethyl-silanyloxymethyl)-phenyl]-/V-[2-(4-hydroxy-2-oxo-2,3- dihydro-benzothiazol-7-yl)-ethyl]-acetamide

The title compound was prepared from [3-(tert-butyl-dimethyl- silanyloxymethyl)-phenyl]-acetic acid and 7-(2-amino-ethyl)-4-hydroxy-3H- benzothiazol-2-one hydrobromide by a similar method to that employed for intermediate 16.

Yield: 5.9 g (90%).

¹H NMR (400 MHz, d₆-DMSO) δ 8.14 (t, J = 5.5 Hz, 1 H), 7.28 (t, J = 7.6 Hz, 1 H), 7.23 - 7.17 (m, 2H), 7.13 (d, J = 7.4 Hz, 1 H), 6.79 (d, J = 8.2 Hz, 1 H),

6.71 (d, J = 8.2 Hz, 1H), 4.72 (s, 2H), 3.40 (s, 2H), 2.73 (s, 2H), 2.63 (t, J =

7.3 Hz, 2H), 0.96 - 0.87 (m, 9H), 0.13 - 0.04 (m, 6H).

Intermediate 23 Carbonic acid 7-(2-{te/t-butoxycarbonyl-[2-(3-hydroxymethyl-phenyl)-ethyl]- amino}-ethyl)-2-oxo-2,3-dihydro-benzothiazol-4-yl ester fert-butyl ester

Borane tetrahydrofuran complex (1 M in THF, 49.1 mL, 49.08 mmol) was added over 15 min to a stirred suspension of 2-[3-(terf-butyl-dimethyl- silanyloxymethyl)-phenyl]-Λ/-[2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)- ethylj-acetamide (5.8 g, 12.27 mmol) in THF (60 mL) at 50 ⁰C. The resulting solution was refluxed for 75 min, cooled to RT, and MeOH was added cautiously until the effervescence ceased (-60 mL). The solvents were removed in vacuo and the residue was taken up in MeOH. Concentrated aqueous HCI (3 mL) was added and the reaction mixture was refluxed for 1 h. The solvents were removed in vacuo to afford title intermediate amino alcohol as a yellow foam (4.93 g, quant.). The residue was taken up in MeOH (50 mL) and triethylamine (6.35 ml, 45.52 mmol) and Boc anhydride (7.27 mL, 31.30 mmol) were added. The resulting solution was stirred at RT for 30 min, then MeOH was removed in vacuo, and the residue was partitioned between EtOAc and water. The organic layer was washed with water, brine, dried (Na₂SO₄), filtered and concentrated in vacuo. The crude product was purified by flash silica chromatography, eluting with hexane/EtOAc (2:1 ) to afford the title compound as a white foam. Yield: 3.18 g (41%).

¹H NMR (400 MHz, d₆-DMSO): δ 7.26 - 7.20 (m, 1 H), 7.1 - 7.11 (m, 2H), 7.10 - 7.04 (m, 1H), 7.04 - 6.93 (m, 2H), 5.16 - 5.09 (m, 1H), 4.51 - 4.44 (m, 2H), OJ

3.38 - 3.33 (m, 2H), 3.31 - 3.23 (m, 2H), 2.7 - 2.65 (m, 4H), 1.52 - 1.46 (m, 9H), 1.37 - 1.24 (m, 9H).

Intermediate 24 Carbonic acid 7-(2-{tert-butoxycarbonyl-[2-(3-formyl-phenyl)-ethyl]-amino}- ethyl)-2-oxo-2,3-dihydro-benzothiazol-4-yl ester tert-butyl ester

The title compound was prepared from carbonic acid 7-(2-{tert- butoxycarbonyl-[2-(3-hydroxymethyl-phenyl)-ethyl]-amino}-ethyl)-2-oxo-2,3-dihydro- benzothiazol-4-yl ester tert-butyl ester by a similar method to that employed for intermediate 17.

Yield: 1.3 g (42%).

LC-MS (Method 6): Rt 2.35 min, m/z 541 [M-H]\

Intermediate 25 {4-[2-(tert-Butyl-dimethyl-silanyloxy)-ethyl]-phenyl}-acetaldehyde

The title compound was synthesised from 2-{4-[2-(te/t-butyl-dimethyl- silanyloxy)ethyl]-phenyl}-ethanol (prepared according to WO9843956) by a similar Dess-Martin oxidation method used in the synthesis of intermediate 14, which was used directly without characterization.

Intermediate 26

Carbonic acid 7-[2-(tert-butoxycarbonyl-{2-[4-(2-hydroxy-ethyl)-phenyl]-ethyl}- amino)-ethyI]-2-oxo-2,3-dihydro-benzothiazol-4-yl ester tert-butyl ester

The title compound was prepared from {4-[2-(terf-butyl-dimethyl-silanyloxy)- ethyl]-phenyl}-acetaldehyde and 7-(2-amino-ethyl)-4-hydroxy-3H-benzothiazol-2-one hydrochloride by similar methods to the reductive amination and Boc-protection steps for Intermediate 7. Prior to bis-Boc protection to afford the title compound, a solution of the intermediate amino TBDMS ether in MeOH (20 ml_) was converted to the amino alcohol by addition of concentrated aqueous HCI (2 ml_). After 20 min the mixture was concentrated in vacuo to afford the crude de-silylated intermediate, which was taken on to react with Boc anhydride to afford the title compound.

¹H NMR (400 MHz, d₆-DMSO, 90 ⁰C): δ 12.02 - 11.94 (m, 1 H), 7.16 - 6.90 (m, 6H), 4.28 - 4.18 (m, 1 H), 3.64 - 3.54 (m, 2H), 3.36 (t, J = 7.0 Hz, 2H), 3.29 (t, J = 7.3 Hz, 2H), 2.76 - 2.64 (m, 6H), 1.49 (s, 9H), 1.33 (s, 9H).

Intermediate 27 Carbonic acid 7-[2-(terf-butoxycarbonyl-{2-[4-(2-oxo-ethyl)-phenyl]-ethyl}- amino)-ethyl]-2-oxo-2,3-dihydro-benzothiazoI-4-yl ester ferf-butyl ester

The title compound was prepared from carbonic acid 7-[2-(teιi- butoxycarbonyl-{2-[4-(2-hydroxy-ethyl)-phenyl]-ethyl}-amino)-ethyl]-2-oxo-2,3- dihydro-benzothiazol-4-yl ester te/t-butyl ester by a similar Dess-Martin oxidation method used in the synthesis of Intermediate 14, and was used directly without characterization.

Intermediate 28

2-[3-(3-Bromo-propoxy)-phenyl]-ethanol o

A solution of 3-(2-hydroxy-ethyl)-phenol (1O g, 72.46 mmol), 1 ,3- dibromopropane (22 ml_, 0.218 mol) and potassium carbonate (16 g, 0.115 mol) in acetone (80 ml_) was warmed to 60 ⁰C overnight. The reaction mixture was cooled, filtered, and then concentrated in vacuo. The residue was purified by by flash silica chromatography, eluting from 25-50% EtOAc/pentane to afford the title compound as a colourless oil. Yield: 16.69 g (89%).

TLC: Rf = 0.33 (25% EtOAc/pentane).

¹H NMR (400 MHz, CDCI₃): δ 7.21 (m, 1H), 6.84 - 6.73 (m, 3H), 4.08 (t, J = 6 Hz, 2H), 3.81 (m, 2H), 3.58 (t, J = 6.5 Hz, 2H), 2.81 (t, J = 6.5 Hz, 2H), 2.29 (m, 2H), 1.81 (br s, 1 H).

Intermediate 29 [3-(3-Bromo-propoxy)-phenyl]-acetaldehyde

Dess-Martin periodinane (25 g, 58.90 mmol) was added to 2-[3-(3-bromo- propoxy)-phenyl]-ethanol (13.8 g, 53.20 mmol) in DCM (300 ml_). After 1 h saturated aqueous NaHCO₃ (25 ml_) and sodium thiosulphate (25 ml_) were added and the mixture was stirred vigorously for 30 minutes, and then separated. The DCM extract was dried (Na₂SO₄), filtered and evaporated in vacuo to afford the crude aldehyde as a white solid. This material was purified by flash silica chromatography, using a gradient from 2.5-10% EtOAc/pentane to afford the title compound as a yellow oil. Yield: 8.5 g (90%).

TLC: Rf = 0.46 (10% EtOAc/pentane). 1H NMR (400 MHz, CDCI₃): δ 9.71 (t, J = 2.4 Hz, 1 H), 7.25 (m, 1 H), 6.86 -

6.70 (m, 3H), 4.08 (t, J = 6 Hz, 2H), 3.62 (d, J = 2.4 Hz, 2H)₁ 3.58 (t, J = 6 Hz, 2H), 2.29 (m, 2H).

Intermediate 30 {2-[3-(3-Bromo-propoxy)-phenyl]-ethyl}-[2-(4-hydroxy-2-oxo-2,3-dihydro- benzothiazol-7-yl)-ethyl]-carbamicacid fe/f-butyl ester

The title compound was prepared from [3-(3-bromo-propoxy)-phenyl]- acetaldehyde and 7-(2-amino-ethyl)-4-hydroxy-3/-/-benzothiazol-2-one hydrochloride by similar methods to those used for intermediate 7. The crude product was purified by HPLC (System 1) to afford the desired product as a white solid.

LC-MS (Method 5): Rt 12.36 min, m/z 551 , 553 [M+H]⁺.

Intermediate 31

(4-{[2-(4-Hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}- thiazol-2-yl)-carbamic acid tert-butyl ester

The title compound was prepared from (4-formylthiazol-2-yl) carbamic acid fert-butyl ester and 7-(2-aminoethyl)-4-hydroxybenzo[d]thiazol-2(3/-/)-one hydrochloride using a similar reductive amination method to that employed for Intermediate 7.

LC-MS (Method 2): Rt 2.21 min, m/z 421 [M-H]^".

Intermediate 32 7-{2-[(2-Amino-thiazol-4-ylmethyl)-amino]-ethyl}-4-hydroxy-3H-benzothiazol-2- one

A solution of (4-formylthiazol-2-yl) carbamic acid te/t-butyl ester (4.67 g, 11.5 mmol) in DCM (13 mL) was treated with TFA (11 ml_). After stirring at RT for 2 h the reaction mixture was concentrated in vacuo and the crude residue was passed down a MeOH-equilibrated SCX-2 cartridge, eluting with DCM/MeOH (1 :1), then eluted with an ammonia solution (2 M in MeOH) to release the desired product, which was used without further purification.

Yield: 3.58 g (quant.). LC-MS (Method 3): Rt 0.38 + 1.54 min, m/z 323 [M+H]⁺.

Intermediate 33

(2-Amino-thiazol-4-ylmethyl)-[2-(4-hydroxy-2-oxo-2,3-dihydro-benzothia zol-7-yl)-ethyl]-carbamic acid tert-butyl ester

The title compound was prepared from (4-formylthiazol-2-yl) carbamic acid te/t-butyl ester by a similar Boc anhydride protection step employed for Intermediate 7. LC-MS (Method 3): Rt 2.29 min, m/z 423 [M-H]^".

Intermediate 34

[2-(2-Bromo-acetylamino)-thiazoI-4-ylmethyl]-[2-(4-hydroxy-2-oxo-2,3-d ihydro-benzothiazol-7-yl)-ethyl]-carbamic acid te/t-butyl ester

The title compound was prepared from (2-amino-thiazol-4-ylmethyl)-[2-(4- hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethyl]-carbamic acid tert

-butyl ester by a similar method to that employed for Intermediate 9. LC-MS (Method 5): Rt 9.17 min, m/z 543, 545 [M-H]^".

intermediate 35 yV-[2-(4-Hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethyl]-2-(4-iodo-phenyl)- acetamide

A solution of (4-iodo-phenyl)-acetic acid (5 g, 19.08 mmol) in MeCN was treated with CDI (3.40 g, 21 mmol) and allowed to stir at RT for 10 min after which a voluminous precipitate was produced. Triethylamine (5.80 ml_, 42 mmol) was added, followed by 7-(2-aminoethyl)-4-hydroxybenzo[d]thiazol-2(3H)-one hydrobromide (5.56 g, 19.08 mmol) and the reaction slurry was stirred at RT for 18 h. The reaction mixture was then partitioned between water (200 ml_) and EtOAc (2 x 300 ml_). The combined organic extracts were washed with 10% aqueous Na₂SO₄ (2 x 100 ml_) and brine (100 ml_), dried (Na₂SO₄), filtered and concentrated in vacuo to afford the title product.

Yield: 6.18 g (71%).

LC-MS (Method 2): Rt 2.98 min, m/z 455 [M+H]\ 496 [M+H+MeCN]⁺.

Intermediate 36 [2-(4-Hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethyl]-[2-(4-iodo-phenyl)- ethylj-carbamic acid terf-butyl ester

The title compound was prepared from 4 Λ/-[2-(4-hydroxy-2-oxo-2,3-dihydro- benzothiazol-7-yl)-ethyl]-2-(4-iodo-phenyl)-acetamide by simitar reduction and Boc- protection methods to those employed for Intermediate 23.

LC-MS (Method 2): Rt 4.15 min, m/z 541 [M+H]⁺, 582 [M+H+MeCN]⁺.

Intermediate 37

{2-[4-(2-Formyl-thiazol-5-yl)-phenyl]-ethyl}-[2-(4-hydroxy-2-oxo-2,3-dihydro- benzothiazol-7-yl)-ethyl]-carbamic acid fert-butyl ester

A solution of [2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethyl]-[2-(4- iodo-phenyl)-ethyl]-carbamic acid fert-butyl ester (850 mg, 1.58 mmol), δ-tributylstannanyl-thiazole^-carbaldehyde (950 mg, 2.36 mmol) and lithium chloride (200 mg, 4.73 mmol) in THF (10 mL) were degassed under argon. PdCI₂(PPh₃)₂ (110 mg, 0.16 mmol) was added and the reaction warmed to 85 ⁰C in a sealed vial for 18 h. The reaction mixture was cooled and stirred with an aqueous solution of KF (10 mL) for 30 min. The solid Bu₃SnF was removed by filtration and the mother liquor was partitioned between water (50 mL) and DCM (2 x 50 mL). The combined organic extracts were washed with water (50 mL) and brine, then dried (MgSO₄), filtered and concentrated in vacuo. The crude product was isolated by careful chromatography on flash silica (250 mL), eluting with 0-2% MeOH/DCM to afford the desired product as a tan solid.

LC-MS (Method 2): Rt 3.93 min, m/z 526 [M+H]⁺.

Intermediate 38

{5-[2-(fert-Butyl-dimethyl-silanyIoxy)-ethyl]-furan-2-yl}-acetic acid methyl ester

^

The title compound was prepared from [5-(2-hydroxy-ethyl)-furan-2-yl]-acetic acid methyl ester by a similar method to that employed for Intermediate 21.

¹H NMR (400 MHz, CDCI₃): δ 6.11 (d, J = 3.1 Hz, 1H), 5.99 (d, J = 3.1 Hz, 1 H), 3.83 (t, J = 6.9 Hz, 2H), 3.72 (s, 3H), 3.64 (s, 2H), 2.81 (t, J = 6.9 Hz, 2H), 0.87 (s, 9H), 0.01 (s, 6H).

LC-MS (Method 6): Rt 2.51 min, m/z 299 [M+H]⁺.

Intermediate 39 {5-[2-(fert-Butyl-dimethyl-silanyloxy)-ethyl]-furan-2-yl}-acetic acid

The title compound was prepared from [5{5-[2-(terf-butyl-dimethyl-silanyloxy)- ethyl]-furan-2-yl}-acetic acid methyl ester by a similar method to that employed for Intermediate 20.

¹H NMR (400 MHz, CDCI₃): δ 6.14 (d, J = 2.8 Hz, 1 H), 6.00 (d, J = 3.1 Hz,

1 H), 3.83 (t, J = 6.8 Hz, 2H), 3.68 (s, 2H), 2.82 (t, J = 6.9 Hz, 2H), 0.87 (s,

9H), 0.01 (s, 6H).

LC-MS (Method 6): Rt 1.72 min, m/z 283 [M-H]^".

Intermediate 40

Carbonic acid 7-[2-(terf-butoxycarbonyl-{2-[5-(2-oxo-ethyl)-furan-2-yl]-ethyl}- amino)-ethyl]-2-oxo-2,3-dihydro-benzothiazol-4-yl ester fert-butyl ester

The title compound was prepared from {5-[2-(tert-butyl-dimethyl-silanyloxy)- ethyl]-furan-2-yl}-acetic acid and 7-(2-aminoethyl)-4-hydroxybenzo[d]thiazol-2(3/^)- one hydrobromide by similar methods to those employed for Intermediates 16, 23 and 25, respectively.

LC-MS (Method 6): Rt 2.17 min, m/z 545 [M-H]^". Intermediate 41

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[3-(4-f ormyl-phenoxy)- propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester

A solution of hyclroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2- methylamino-bicyclo[2.2.1]hept-7-yl ester (Intermediate 6) (200 mg, 0.55 mmol), 4-(3- bromo-propoxy)-benzaldehyde (200 mg, 0.83 mmol) and diisopropylethylamine (290 μL, 1.65 mmol) in MeCN was warmed to 65 ⁰C for 72 h. The reaction mixture was concentrated in vacuo and then purified by flash chromatography on a silica-gel cartridge (20 g lsolute cartridge), eluting with 0-1% MeOH/DCM to afford the desired product as an amber oil. LC-MS (Method 2): Rt 2.53 min, m/z 526.33 [M+H]⁺.

Intermediate 42

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[3-(4-{[(R)-2-(tert-butyl- dimethyl-silanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydro-quinoIin-5-yl)-ethylamino]- methyl}-phenoxy)-propyl]-methyl-arnino}-bicyclo[2.2. 1]hept-7-yl ester

A solution of hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-{[3-(4- formyl-phenoxy)-propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester (Intermediate 41) (178 mg, 0.339 mmol) and 5-[(R)-2-amino-1-(te/t-butyl-dimethyl-silanyloxy)-ethyl]-8- hydroxy-1 /-/-quinolin-2-one (158 mg, 0.475 mmol) (prepared according to WO2007/102771) in NMP was stirred at RT for 30 min. Sodium triacetoxyborohydride (200 mg, 0.98 mmol) was added to the reaction mixture and the stirring continued at RT for 18 h. The reaction mixture was quenched by the addition of saturated aqueous NaHCO₃ prior to extracting with EtOAc. The phases were separated and the organic layer was washed successively with water and brine, dried (NaSO₄), and then concentrated in vacuo. The crude product was purified by flash chromatography (20 g lsolute cartridge), eluting from 0-5% MeOH/DCM to afford the title compound as an off-white solid.

LC-MS (Method 2): Rt 2.55 min, m/z 844 [M+H]⁺.

Intermediate 43

9-Hydroxy-9H-xanthene-9-carboxylic acid (1 S,2S,4R,7R)-2-methylamino- bicyclo[2.2.1]hept-7-yl ester

The title compound was prepared from ((1 S,2S,4R,7R)-7-hydroxy- bicyclo[2.2.1]hept-2-yl)-methyl-carbamic acid-tert-butyl ester and 9-hydroxy-9H- xanthene-9-carboxylic acid methyl ester (prepared according to US 2005/0025718) by similar methods to those employed for Intermediates 5 and 6, respectively. LC-MS (Method 2): Rt 2.24 min, m/z 366 [M+H]⁺.

Intermediate 44

9-Hydroxy-9H-xanthene-9-carboxylic acid (1 S,2S,4R,7R)-2-{[3-(4-formyl- phenoxy)-propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester

The title compound was prepared from 9-hydroxy-9/-/-xanthene-9-carboxylic acid (1S,2S,4R,7R)-2-{[3-(4-formyl-phenoxy)-propyl]-methyl-amino}- bicyclo[2.2.1]hept-7-yl ester and 4-(3-bromo-propoxy)-benzaldehyde by a similar method to that employed for Intermediate 41.

LC-MS (Method 4): Rt 2.54 min, m/z 528 [M+H]⁺.

Intermediate 45 θ-Hydroxy-ΘH-xanthene-g-carboxylic acid (1S₎2S,4R,7R)-2-{[3-(4-{[(R)-2-(fert- butyl-dimethyl-silanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydro-quinolin-5-yl)- ethylamino]-methyl}-phenoxy)-propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester

The title compound was prepared from 9-hydroxy-9H-xanthene-9-carboxylic acid (1S,2S,4R,7R)-2-methylamino-bicyclo[2.2.1]hept-7-yl ester and 5-[(R)-2-amino- 1-(tert-butyl-dimethyl-silanyloxy)-ethyl]-8-hydroxy-1 H-quinolin-2-one by a similar method to that employed for Intermediate 42.

LC-MS (Method 4): Rt 2.42 min, m/z 846 [M+Hf.

Intermediate 46

Hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-amino-bicyclo[2.2.1]hept- 7-yl ester

The title compound was prepared from (1 R,4R,7R)-7-benzyloxy- bicyclo[2.2.1]heptan-2-one and benzylamine by similar methods to those employed for Intermediates 3-6.

LC-MS (Method 2): Rt 2.32 min, m/z 350 [M+H]⁺.

Intermediate 47 Hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-[3-(4-formyl-phenoxy)- propylamino]-bicyclo[2.2.1]hept-7-yl ester

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid

(1 S,2S,4R,7R)-2-amino-bicyclo[2.2.1]hept-7-yl ester by a similar method to that employed for Intermediates 41.

LC-MS (Method 2): Rt 2.54 min, m/z 512 [M+H]⁺.

Intermediate 48

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[3-(2-hydroxy-ethyl)- benzyl]-methyl-amino}-bicyclo[2.2.1 ]hept-7-yl ester

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-methylamino-bicyclo[2.2.1]hept-7-yl ester (Intermediate 6) and 2-(3- bromomethyl-phenyl)-ethanol (prepared according EP 472449) by a similar method to that employed for Intermediate 41.

LC-MS (Method 6): Rt 2.18 min, m/z 498 [M+H]⁺.

Intermediate 49 4-Hydroxymethyl-/V-(2-oxo-ethyl)-benzamide

The title compound was prepared from Λ/-(2,2-dimethoxy-ethyl)-4- hydroxymethyl-benzamide (Intermediate 16) by a similar method to that employed for Intermediate 15.

LC-MS (Method 2): Rt 1.80 min, m/z 194 [M+H]⁺.

Intermediate 50 Hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-{[2-(3-hydroxymethyl- benzoylamino)-ethyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester

A mixture of hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2- methylamino-bicyclo[2.2.1]hept-7-yl ester (200 mg, 0.55 mmol) (Intermediate 6), 4-hydroxymethyl-Λ/-(2-oxo-ethyl)-benzamide (1 17 mg, 0.61 mmol) and sodium triacetoxy borohydride (128 mg, 0.61 mmol) in dry NMP (5 ml.) were stirred at RT for 18 h. The reaction mixture was quenched by the addition of saturated aqueous NaHCO₃ prior to extracting with EtOAc. The phases were separated and the organic layer was washed successively with water and brine, dried (NaSO₄), and then concentrated in vacuo. The crude product was passed down a MeOH equilibrated SCX-2 cartridge and eluted with an ammonia solution (2 M in MeOH) to release the product. Further purification was carried out by flash column chromatography over silica, eluting with 0-2.5% MeOH/DCM to afford the title compound as a white foam.

Yield: 135 mg (46%).

LC-MS (Method 2): Rt 2.44 min, m/z 541 [M+H]⁺.

Intermediate 51

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(3-formyl- benzoylamino)-ethyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-{[2-(3-hydroxymethyl-benzoylamino)-ethyl]-methyl-amino}- bicyclo[2.2.1]hept-7-yl ester by a similar a method to that employed for Intermediate 17.

LC-MS (Method 2): Rt 2.56 min, m/z 539 [M+H]⁺.

Intermediate 52

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-(fert-butyl- dimethyl-silanyloxy)-2-(8-hydroxy-2-oxo-1 ,2-dihydro-quinolin-5-yl)-ethylamino]- methyl}-benzoylamino)-ethyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester

A mixture of hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-{[2-(3- formyl-benzoylamino)-ethyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester (48 mg, 0.089 mmol), 5-[(R)-2-amino-1-(tert-butyl-dimethyl-silanyloxy)-ethyl]-8-hydroxy-1 H- quinolin-2-one (36 mg, 0.107 mmol) and 3 A molecular sieves were stirred in anhydrous MeOH overnight and then treated with sodium triacetoxyborohydride (amount?). After 20 min the reaction mixture was quenched by the addition of saturated aqueous NaHCO₃ prior to extracting with EtOAc. The phases were separated and the organic layer was washed successively with water and brine, dried (NaSO₄), and then concentrated in vacuo. The crude product was purified by flash chromatography on a silica-gel cartridge (20 g lsolute cartridge), eluting with 0-5% MeOH/DCM to afford the title compound as an off-white solid. LC-MS (Method 2): Rt 2.50 min, m/z 857 [M+H]⁺.

Intermediate 53

3-{[(1S,2S,4R,7R)-7-(2-Hydroxy-2,2-di-thiophen-2-yl-acetoxy)-bicyclo[2.2.1]hept- 2-yl]-methyl-amino}-propionic acid

A solution of hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2- methylamino-bicyclo[2.2.1]hept-7-yl ester (166 mg, 0.45 mmol) (Intermediate 6) and acrylic acid (50 μl_, 0.606 mmol) in DCM (5 ml_) in a sealed tube was warmed to 50 ⁰C for 60 h. The reaction mixture was cooled and concentrated in vacuo, and then 'chased' with toluene to afford the title compound as a white solid.

LC-MS (Method 2): Rt 2.38 min, m/z 436 [M+H]⁺.

Intermediate 54

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2R,4S,7S)-7-{[2-(4-formyl- phenylcarbamoyl)-ethyl]-methyl-amino}-bicyclo[2.2.1]hept-2-yl ester

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid (1S,2R,4S,7S)-7-methylamino-bicyclo[2.2.1]hept-2-yl ester (Intermediate 6) and (4-amino-phenyl)-methanol by similar methods to those employed for Intermediates 16 and 17, respectively.

LC-MS (Method 2): Rt 2.59 min, m/z 539 [M+H]⁺.

Intermediate 55

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-(tert-butyl- dimethyl-silanyloxy)-2-(8-hydroxy-2-oxo-1,2-dihydro-quinolin-5-yl)-ethylamino]- methyl}-phenylcarbamoyl)-ethyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid (1 S,2R,4S,7S)-7-{[2-(4-formyl-phenylcarbamoyl)-ethyl]-methyl-amino}- bicyclo[2.2.1]hept-2-yl ester and 5-[(R)-2-amino-1-(tert-butyl-dimethyl-silanyloxy)- ethyl]-8-hydroxy-1 H-quinolin-2-one by a similar method to that employed for Intermediate 52.

LC-MS (Method 2): Rt 2.55 min, m/z 857 [M+H]⁺.

Intermediate 56

3-(2-Hydroxy-ethyl)-benzoic acid methyl ester

2-(3-Bromo-phenyl)-ethanol (5.00 g, 24.87 mmol), 1 ,1'-bis(diphenylphosphino) ferrocene-palladium dichloride (0.70 g, 0.85 mmol) and triethylamine (7.00 g, 69.18 mmol) in MeOH (30 ml.) was heated at 90 ⁰C (internal temperature) over a period of 10 h under carbon monoxide atmosphere (5 bar). After cooling to RT the solution was concentrated in vacuo and then partitioned between ethyl acetate and water. The organic layer was washed with brine and evaporated in vacuo. The crude product was purified by flash silica chromatography eluting with ethyl acetate//-hexane (1 :1).

Yield: 3.3 g (74%).

¹H NMR (400 MHz, CDCI₃): δ 7.93 - 7.90 (m, 2H), 7.44 (d, J = 7.2 Hz, 1 H),

7.39 (t, J = 8.1 Hz, 1 H), 3.92 (s, 3H), 3.89 (t, J = 6.2 Hz, 2H), 2.93 (t, J = 6.4

Hz, 2H), hydroxyl proton not observed.

Intermediate 57 3-(2-Hydroxy-ethyl)-benzoic acid

The title compound was prepared from 3-(2-hydroxy-ethyl)-benzoic acid methyl ester by a similar a method to that employed for Intermediate 20. LC-MS (Method 2): Rt 2.08 min, m/z 165 [M-H]^".

Intermediate 58

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-[(2-amino-ethyl)-methyl- amino]-bicyclo[2.2.1]hept-7-yl ester

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-[(2-amino-ethyl)-methyl-amino]-bicyclo[2.2.1]hept-7-yl ester and (2-oxo-ethyl)-carbamic acid tert-butyl ester by similar methods to those employed for Intermediates 48 and 6, respectively.

LC-MS (Method 9): Rt 1.97 min, m/z 407 [M+H]⁺.

Intermediate 59

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-({2-[3-(2-hydroxy-ethyl)- benzoylamino]-ethyl}-methyl-amino)-bicyclo[2.2.1]hept-7-yl ester

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-[(2-amino-ethyl)-methyl-amino]-bicyclo[2.2.1]hept-7-yl ester and 3-(2-hydroxy-ethyl)-benzoic acid (Intermediate 57) by a similar method to that employed for Intermediate 16.

LC-MS (Method 3): Rt 2.59 min, m/z 555 [M+H]⁺.

Intermediate 60 8-Acetyl-5-benzyloxy-4W-benzo[1 ,4]oxazin-3-one

Chloroacetyl chloride (1.25 ml_, 15.5 mmol) was added to a stirred mixture of 1-(3-amino-4-benzyloxy-2-hydroxy-phenyl)-ethanone (3.63 g, 14.1 mmol) and

NaHCO₃ (2.69 g, 32.0 mmol) in dry DMF (20 ml_). The reaction mixture was stirred for 2 h at RT. Caesium carbonate (5.53 g, 17.0 mmol) was added and the mixture was heated at 100 ⁰C, protected from moisture, for 2.5 h. The reaction mixture was cooled to RT and poured into water (-300 ml_) to afford a turbid, milky solution from which a solid precipitated. The solid was removed by vacuum filtration and taken up in EtOAc by sonication and the aqueous layer was extracted with EtOAc. The combined organic layers were further washed with water, brine, dried (MgSO₄), and concentrated in vacuo to afford a dark brown, fine solid.

Yield: 3.60 g (86%). LC-MS (Method 4): Rt 3.24 min, m/z 298 [M+H]⁺.

intermediate 61 5-Benzyloxy-8-(2-chloro-acetyl)-4H-benzo[1,4]oxazin-3-one

Benzyltrimethylammonium dichloroiodate (9.27 g, 26.6 mmol) was added to a stirred solution of 8-acetyl-5-benzyloxy-4/-/-benzo[1 ,4]oxazin-3-one (3.60 g, 12.1 mmol) in a mixture of DCM (65 ml_), acetic acid (22 ml_), and water (3.6 ml_), which was heated at 65 ⁰C, protected from light, for 18 h. The reaction mixture was cooled to RT and a solution of sodium bisulphite (3.78 g) in water (65 ml_) was added and mixed vigorously to form a suspension. After stirring for 0.5 h diethyl ether (150 ml_) was added and the solids collected by filtration, washed with water, then ether and dried at 40 ⁰C in vacuo to afford a light brown crystalline solid. Yield: 3.56 g (89%).

LC-MS (Method 4): Rt 3.50 min, m/z 332 [M+H]⁺.

Intermediate 62 5-Benzyloxy-8-((R)-2-chloro-1-hydroxy-ethyl)-4W-benzo[1,4]oxazin-3-one

To a suspension of 5-benzyloxy-8-(2-chloro-acetyl)-4H-benzo[1 ,4]oxazin-3- one (8.38 g, 25.3 mmol) in dry THF (150 mL) under nitrogen was added (R)-(+)-CBS (5.0 mL, 5.0 mmol, 1 M in toluene) and the mixture was cooled to -15 °C. Borane- THF complex (28 mL, 28 mmol, 1 M in THF) was slowly added via syringe pump over 2 h, after which the reaction mixture had become a clear solution with some solids present. The reaction was quenched by addition of MeOH (12 mL) (gas evolved) and was then allowed to warm to RT. After this time a beige, crystalline material precipitated, which was removed by filtration. This was found to be unreacted starting material. The filtrate was partitioned between water and EtOAc, and the water layer was further extracted with EtOAc. The combined organic layers were washed with brine, dried (MgSO₄), and concentrated in vacuo to afford a beige, sticky solidified gum. This material was dissolved in DCM and some insoluble white solids were removed by vacuum filtration, which were found to be CBS by-products. The filtrate was concentrated and purified by chromatography over silica gel on a Teledyne ISCO CombiFlash Companion (120 g cartridge) eluting with 0-20% EtOAc/DCM to afford the title product as an off-white solid. Analysis by chiral HPLC (Chiralpak IA, 15% IPA/heptane, 1 mL/min) revealed 90% e.e. of the first eluting enantiomer (Rt = 21.0 min) (Second enantiomer: Rt = 24.5 min). A single crystallisation from refluxing EtOAc (35 ml_)/heptane (20 mL) afforded the title compound as small colourless crystals in 97% e.e. as determined by chiral HPLC analysis.

Yield: 4.76 g (59%).

LC-MS (Method 4): Rt 3.16 min, m/z 332 [M-H]^".

Intermediate 63 5-Benzyloxy-8-[(R)-1-(te/t-butyl-dimethyl-silanyloxy)-2-chloro-ethyl]-4H- benzo[1 ,4]oxazin-3-one

2,6-Lutidine (3.4 mL, 29 mmol) was added to a solution of 5-benzyloxy-8-((R)-

2-chloro-1-hydroxy-ethyl)-4H-benzo[1 ,4]oxazin-3-one (4.76 g, 14.3 mmol) in dry DMF (30 mL) at 0 ^CC under a nitrogen atmosphere, followed by slow addition of TBDMS trifluoromethanesulfonate (6.4 mL, 28 mmol). Fumes were formed and the reaction mixture was stirred below 10 ⁰C for 5 min, after which the reaction mixture was allowed to warm to RT. After 45 minutes MeOH (3.0 mL) was added cautiously and the reaction mixture was poured onto water to afford a white suspension. This was extracted with EtOAc (2x) and the combined organic layers were washed with water, brine, dried (MgSO₄), and concentrated in vacuo to afford the title compound as a white solid, which was used without further purification Yield: 6.34 g (99%).

LC-MS (Method 4): Rt 5.14 min, molecular ion no observed. Intermediate 64

8-[(R)-2-Azido-1-(tert-butyl-dimethyl-silanyloxy)-ethyl]-5-benzyloxy-4H- benzo[1 ,4]oxazin-3-one

A mixture of 5-benzyloxy-8-[(R)-1-(tø/t-butyl-dimethyl-silanyloxy)-2-chloro- ethyl]-4H-benzo[1 ,4]oxazin-3-one (3.00 g, 5.7 mmol), sodium azide (2.28 g, 35.1 mmol), and potassium iodide (5.85 g, 35.2 mmol) in dry DMF (24 ml_) was heated at 120 ⁰C under a nitrogen atmosphere. The reaction mixture became dark brown very quickly. After heating for 72 h the reaction mixture was cooled to RT and poured onto water. The solids were collected by filtration and washed with water. This material was then taken up in EtOAc, dried over MgSO₄, filtered, and concentrated in vacuo. The crude product was purified over a short silica column to remove baseline material, eluting with 20-25% EtOAc/cyclohexane, and afforded the title compound as a white solid.

Yield: 1.2O g (46%).

¹H NMR (400 MHz, CDCI₃): δ 7.77 (br. s, 1 H), 7.45 - 7.35 (m, 5H), 7.14 (d, J = 8.8 Hz, 1 H)₁ 6.69 (d, J = 8.8 Hz, 1 H), 5.13 (m, 1 H), 5.08 (m, 2H), 4.48 (m, 2H)₁ 3.25 (m, 2H), 0.92 (s, 9H), -0.04 (s, 6H).

Intermediate 65

8-[(R)-2-Amino-1-(tert-butyl-dimethyl-silanyloxy)-ethyl]-5-hydroxy-4H- benzo[1 ,4]oxazin-3-one

A solution of 8-[(R)-2-azido-1-(tert-butyl-dimethyl-silanyloxy)-ethyl]-5- benzyloxy-4/-/-benzo[1 ,4]oxazin-3-one (1.20 g, 2.64 mmol) in EtOAc/EtOH (15 ml_, 1 :1) in the presence of a catalytic amount of Pd/C was stirred under a hydrogen atmosphere at RT for 48 h. The reaction mixture was filtered over Celite, washed with EtOH/EtOAc, and the filtrate was concentrated in vacuo to afford the title compound as a light brown foam, which was used without further purification.

Yield: 892 mg (quant.).

LC-MS (Method 4): Rt 2.33 min, m/z 339 [M+H]⁺.

Compounds 1-12 were also prepared:

Compound 1

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[2-(4-hydroxy-2- oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}-phenylcarbamoyl)- ethyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester trifluoroacetate

An aliquot (0.29 ml_) of a solution of (4-acryloylamino-benzyl)-[2-(4-hydroxy-2- oxo-2,3-dihydro-benzothiazol-7-yl)-ethyl]-carbamic acid tert-butyl ester (3.8 mmol) (Intermediate 10) in NMP (10 ml.) and acetic acid (0.23 mL) was added to a solution of hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-methylamino- bicyclo[2.2.1]hept-7-yl ester (Intermediate 6) (0.075 mmol) in NMP (0.1 mL). The reaction was mixed for 5 days at 70 ⁰C. MeOH (0.5 mL) was added and the solution was passed down a MeOH equilibrated TsOH-65 resin cartridge (0.2 g) and eluted with an ammonia solution (2 M in MeOH) to release the crude Boc-intermediate after concentration in vacuo. Formic acid (0.5 mL) was added was added to the crude reaction mixture and the solution was stirred at RT for 2 h. The reaction mixture was concentrated in vacuo, dissolved in DMSO (0.4 mL), and then purified by preparative reversed-phase HPLC (System 2) to afford the title compound as a white solid. Yield: LC-MS (Method 8): Rt 1.37 min, m/z 734 [M+H]⁺. Compound 2

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{2-[2-(4-hydroxy-2- oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-ethyl}-benzoylamino)-ethyl]- methyl-amino}-bicyclo[2.2.1]hept-7-yl ester trifluoroacetate

[2-(4-Hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethyl]-{2-[4-(2-oxo- ethylcarbamoyl)-phenyl]-ethyl}-carbamic acid tert-butyl ester (3.8 mmol) (Intermediate 15) was dissolved in NMP (10 mL) and acetic acid (0.23 ml) was added. An aliquot of this solution (0.29 mL) was added to a reaction vessel containing hydroxy-di- thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-methylamino-bicyclo[2.2.1]hept-7-yl ester (0.075 mmol) (Intermediate 6) in NMP (0.1 mL). The reaction was stirred for 5 min and a sodium triacetoxyborohydride (0.15 mL of a 1.14 M solution in NMP (0.17 mmol)) was added and the reaction mixture was stirred for 16 h. MeOH (0.5 mL) was added and the solution was passed down a MeOH equilibrated TsOH-65 resin cartridge (0.2 g) and eluted with an ammonia solution (2 M in MeOH) to afford the crude Boc-intermediate after concentration in vacuo. Formic acid (0.5 mL) was added to the crude reaction mixture and the solution was stirred at RT for 2 h. The reaction mixture was concentrated in vacuo, dissolved in DMSO (0.4 mL), and then purified by preparative reversed-phase HPLC (System 2) to afford the title compound as a white solid.

LC-MS (Method 8): Rt 1.40 min, m/z 748 [M+H]⁺.

Compounds 3-7, shown in the table below, were prepared using a similar method to that described for Compound 2 using the appropriate β₂-substituted aldehyde intermediates and hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2- methylamino-bicyclo[2.2.1]hept-7-yl ester (Intermediate 6) to afford the title compounds as TFA salts (analysis undertaken using LC-MS Method 8).

Compound 8

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[(4-{[2-(4-hydroxy-2-oxo-

2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}-phenylcarbamoyl)-methyl]- methyl-amino}-bicyclo[2.2.1]hept-7-yl ester trifluoacetate

[4-(2-Bromo-acetylamino)-benzyl]-[2-(4-hydroxy-2-oxo-2,3-dihydro- benzothiazol-7-yl)-ethyl]-carbamic acid te/t-butyl ester (1.2 mmol) (Intermediate 9) was dissolved in NMP (4 ml_) and an aliquot (100 μl_) of this solution was added to the reaction vessel containing hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2- methylamino-bicyclo[2.2.1]hept-7-yl ester (Intermediate 6) (0.025 mmol) and triethylamine (0.066 mmol) in NMP (200 μL). The reaction mixture was allowed to stand at RT for 6 h. MeOH (0.6 mL) was added and the solution was passed down a MeOH equilibrated TsO-H65 resin (0.2 g) and eluted with an ammonia solution (3.5 M in MeOH) to release the Boc-protected intermediate. Formic acid (0.5 mL) was added was added to the crude reaction mixture and the solution was stirred at RT for 2 h. The reaction mixture was concentrated in vacuo, dissolved in DMSO (0.4 mL), and then purified by preparative reversed-phase HPLC (System 2) to afford the title compound as a white solid.

Yield:

LC-MS (Method 8): Rt 1.41 min, m/z 720 [M+H]⁺.

Compounds 9-10, shown in the table below, were prepared using a similar method to that described for Compound 8 using the appropriate β₂-substituted bromo acetamide intermediates and hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2- methylamino-bicyclo[2.2.1]hept-7-yl ester (Intermediate 6) to afford the title compounds as TFA salts (analysis undertaken using LC-MS Method 8).

Compound 11 Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[3-(3-{2-[2-(4-hydroxy-2- oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-ethyl}-phenoxy)-propyl]-methyl- amino}-bicyclo[2.2.1]hept-7-yl ester trifluoroacetic acid

{2-[3-(3-Bromo-propoxy)-phenyl]-ethyl}-[2-(4-hydroxy-2-oxo-2,3-dihydro- benzothiazol-7-yl)-ethyl]-carbamic acid terf-butyl ester (1.95 mmol) (Intermediate 30) and triethylamine (0.404 g) were dissolved in sufficient NMP to give 2 mL of solution. An aliquot (50 μL) of this solution was added to a solution of hydroxy-di-thiophen-2-yl- acetic acid (1S,2S,4R,7R)-2-methylamino-bicyclo[2.2.1]hept-7-yl ester (Intermediate 6) (0.05 mmol) in NMP (100 μL). The reaction mixture was heated at 85 ⁰C for 18 h. MeOH (0.7 mL) was added and the solution was passed down a MeOH equilibrated TsO-H65 resin (0.2 g) and eluted with an ammonia solution (3.5 M in MeOH) to release the Boc-protected intermediate. The fractions containing the product were evaporated. Formic acid (0.5 mL) was added to the crude reaction mixture and the solution was stirred at RT for 2 h. The reaction mixture was concentrated in vacuo, dissolved in DMSO (0.4 mL), and then purified by preparative reversed-phase HPLC (System 2) to afford the title compound as a white solid. LC-MS (Method 8): Rt 1.44 min, m/z 735 [M+H]⁺.

Compound 12

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[3-(4-{[2-(4-hydroxy-2- oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}-phenoxy)-propyl]- methyl-amino}-bicyclo[2.2.1]hept-7-yl ester formate

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[3-(4-formyl-phenoxy)-propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester (Intermediate 41) and 7-((R)-2-amino-1-hydroxy-ethyl)-4-hydroxy-3H- benzothiazol-2-one hydrochloride (prepared according to WO2007/027133) by a similar a method to that employed in intermediate 42. The crude product was purified by preparative reversed-phase HPLC (System 1), eluting with 10-30% MeCN/water (0.1% formic acid) over 20 min. The pure fractions were combined and lyophilized to afford the title product as white solid.

LC-MS (Method 5): Rt 5.55 min, m/z 720 [M+H]⁺.

The invention will now be illustrated, but not limited, by the following Examples:

Example 1

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[3-(4-{[(R)-2-hydroxy-2-(4- hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}-phenoxy)- propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester formate

To a solution of hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-{[3-(4- formyl-phenoxy)-propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester (Intermediate 41) (178 mg, 0.34 mmol) and 7-((R)-2-amino-1-hydroxy-ethyl)-4-hydroxy-3H- benzothiazol-2-one (97 mg, 0.34 mmol) (prepared according to WO 2007027134) in NMP (10 ml_) was added sodium triacetoxyborohydride (144 mg, 0.68 mmol). The resulting solution was stirred at RT for 18 h. Further sodium triacetoxyborohydride (72 mg, 0.34 mmol) was added and the stirring continued for 18 h. The reaction mixture was poured into water and extracted into EtOAc. The organic extracts were washed successively with water and brine, dried (Na₂SO₄), filtered and concentrated in vacuo. The crude product was purified by preparative reversed-phase HPLC (Method 1), eluting with 10-30% MeCN/water (+0.1% formic acid) over 20 min. The pure fractions were combined and lyophilized to afford the title product, a cream solid, as a formate salt.

Yield: 138 mg (56%).

LC-MS (System 1 ): Rt 5.91 min, m/z 736 [M+H]⁺.

Example 2 Hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-{[3-(4-{[(R)-2-hydroxy-2-(8- hydroxy-2-oxo-1 ,2-dihydro-quinolin-5-yl)-ethylamino]-methyl}-phenoxy)- propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester formate

A solution of hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-{[3-(4- {[(R)-2-(tert-butyl-dimethyl-silanyloxy)-2-(8-hydroxy-2-oxo-1 ,2-dihydro-quinolin-5-yl)- ethylamino]-methyl}-phenoxy)-propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester (78 mg, 0.092 mmol) (Intermediate 42) in THF (3 mL) under nitrogen was treated with triethylamine trihydrofluoride (37 μL, 0.23 mmol). After stirring at RT overnight, the reaction mixture was neutralised with saturated aqueous NaHCO₃ and extracted with DCM. The combined organic extracts were washed with brine, dried (Na₂SO₄), filtered, and concentrated in vacuo. The crude product was purified by preparative reversed-phase HPLC (System 1), eluting with 10-30% MeCN/water (0.1% formic acid) over 20 min. The pure fractions were combined and lyophilized to afford the title product as white solid. Yield: 38 mg (56%).

LC-MS (Method 5): Rt 5.34 min, m/z 730 [M+H]⁺.

Example 3 θ-Hydroxy-ΘH-xanthene-θ-carboxylic acid (1 S,2S,4R,7R)-2-{[3-(4-{[(R)-2- hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]- methyl}-phenoxy)-propyl]-methyl-amino}-bicyclo[2.2.1 ]hept-7-yl ester formate

The title compound was prepared from 9-hydroxy-9H-xanthene-9-carboxylic acid (1S,2S₎4R,7R)-2-{[3-(4-formyl-phenoxy)-propyl]-methyl-amino}- bicyclo[2.2.1]hept-7-yl ester (Intermediate 44) and 7-((R)-2-amino-1-hydroxy-ethyl)-4- hydroxy-3/-/-benzothiazol-2-one hydrochloride by a similar a method to that employed for Intermediate 52. The crude product was purified by preparative reversed-phase HPLC (System 1), eluting with 10-25% MeCN/water (0.1% formic acid) over 20 min The pure fractions were combined and lyophilized to afford the title product as white solid. LC-MS (Method 5): Rt 5.83 min, m/z 738 [MH-H]⁺.

Example 4

9-Hydroxy-9H-xanthene-9-carboxylic acid (1 S,2S,4R,7R)-2-{[3-(4-{[(R)-2- hydroxy-2-(8-hydroxy-2-oxo-1,2-dihydro-quinolin-5-yl)-ethylamino]-methyl}- phenoxy)-propyl]-methyI-amino}-bicyclo[2.2.1]hept-7-yl ester formate

The title compound was prepared from 9-hydroxy-9H-xanthene-9-carboxylic acid (1S,2S,4R,7R)-2-{[3-(4-{[(R)-2-(terf-butyl-dimethyl-silanyloxy)-2-(8-hydroxy-2- oxo-1 ,2-dihydro-quinolin-5-yl)-ethylamino]-methyl}-phenoxy)-propyl]-methyl-amino}- bicyclo[2.2.1]hept-7-yl ester (Intermediate 45) by a similar a method to that employed for Example 2. The crude product was purified by preparative reversed-phase HPLC (System 1), eluting with 10-30% MeCN/water (0.1% formic acid) over 20 min. The pure fractions were combined and lyophilized to afford the title product as white solid.

LC-MS (Method 5): Rt 5.65 min, m/z 732 [M+H]⁺.

Example 5

9-Hydroxy-9H-xanthene-9-carboxylic acid (1 S,2S,4R,7R)-2-{[3-(4-{[(R)-2- hydroxy-2-(5-hydroxy-3-oxo-3,4-dihydro-2W-benzo[1,4]oxazin-8-yI)-ethylamino]- methyl}-phenoxy)-propyl]-methyl-amino}-bicyclo[2.2.1 ]hept-7-yl ester formate

The title compound was prepared from 9-hydroxy-9H-xanthene-9-carboxylic acid (1 S,2S,4R,7R)-2-{[3-(4-formyl-phenoxy)-propyl]-methyl-amino}- bicyclo[2.2.1]hept-7-yl ester (Intermediate 44) by a similar a method to that employed for Intermediate 42 and Example 2, respectively. The crude product was purified by preparative reversed-phase HPLC (System 1), eluting with 10-30% MeCN/water (0.1% formic acid) over 20 min. The pure fractions were combined and lyophilized to afford the title product as white solid. LC-MS (Method 5): Rt 5.64 min, m/z 736 [M+H]⁺.

Example 6

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-[3-(4-{[(R)-2-hydroxy-2-(4- hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}-phenoxy)- propylamino]-bicyclo[2.2.1]hept-7-yl ester formate

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid

(1 S,2S,4R,7R)-2-[3-(4-formyl-phenoxy)-propylamino]-bicyclo[2.2.1]hept-7-yl ester (Intermediate 47) and 7-((R)-2-amino-1-hydroxy-ethyl)-4-hydroxy-3H-benzothiazol-2- one hydrochloride by a similar method to that employed for Intermediate 52. The crude product was purified by preparative reversed-phase HPLC (System 1), eluting with 10-30% MeCN/water (0.1% formic acid) over 20 min. The pure fractions were combined and lyophilized to afford the title product as white solid. LC-MS (Method 5): Rt 5.89 min, m/z 722 [M+H]⁺.

Example 7 Hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-[(3-{2-[(R)-2-hydroxy-2-(4- hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-ethyl}-benzyl)- methyl-amino]-bicyclo[2.2.1]hept-7-yl ester acetate

To a solution of hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-{[3-(2- hydroxy-ethyl)-benzyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester (Intermediate 48) (174 mg, 0.35 mmol) in DCM (1 mL) at 0 ⁰C was added TFA (27 μl_, 0.35 mmol). The resulting solution was stirred for 2 min before the addition of Dess-Martin periodinane (178 mg, 0.42 mmol) and stirring was continued for 20 min. The reaction mixture was treated with a mixture of aqueous saturated NaHCO₃ and sodium thiosulphate solution, followed by EtOAc and was then stirred vigorously for 10 min. The organic layer was separated and washed with saturated aqueous NaHCO₃, brine, dried over Na₂SO₄, filtered, and concentrated in vacuo. The residue was re-dissolved in MeOH (1 mL) and 7-((R)-2-amino-1-hydroxy-ethyl)-4-hydroxy-3/-/-benzothiazol-2-one hydrochloride (101 mg, 0.38 mmol) and acetic acid (20 μl_, 0.35 mmol) were added. The reaction mixture was cooled to 0 ⁰C and stirred for 5 min prior to addition of sodium cyanoborohydride (33 mg, 0.52 mmol). The reaction mixture was stirred at RT for 1 h. The solvent was removed in vacuo and the residue was purified by preparative reversed-phase HPLC (Xbridge column 50 mm x 19 mm - T9540A), eluting with increasing gradients of MeCN in water (+0.1% ammonium acetate. The pure fractions were combined and lyophilized to afford the title product as white solid. LC-MS (Method 7): Rt 2.71 min, m/z 706 [M+H]⁺.

Example 8

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-hydroxy-2-(4- hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}- benzoylamino)-ethyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-{[2-(3-formyl-benzoylamino)-ethyl]-methyl-amino}-bicyclo[2.2.1]hept- 7-yl ester (Intermediate 51) and 7-((R)-2-amino-1-hydroxy-ethyl)-4-hydroxy-3H- benzothiazol-2-one hydrochloride by a similar method to that employed for Intermediate 52. The crude product was purified by preparative reversed-phase HPLC (System 1), eluting with 10-25% MeCN/water (0.1% formic acid) over 15 min. The pure fractions were combined and lyophilized to afford the title product as white fluffy solid.

LC-MS (Method 5): Rt 5.51 min, m/z 749 [M+H]⁺.

Example 9

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-hydroxy-2-(8- hydroxy-2-oxo-1,2-dihydro-quinolin-5-yl)-ethylamino]-methyl}-benzoylamino)- ethyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester formate

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-{[2-(4-{[(R)-2-(te/t-butyl-dimethyl-silanyloxy)-2-(8-hydroxy-2-oxo-1 ,2- dihydro-quinolin-5-yl)-ethylamino]-methyl}-benzoylamino)-ethyl]-methyl-amino}- bicyclo[2.2.1]hept-7-yl ester (Intermediate 52) by a similar method to that employed for Example 2. The crude product was purified by preparative reversed-phase HPLC (System 1), eluting with 10-25% MeCN/water (0.1% formic acid) over 15 min. The pure fractions were combined and lyophilized to afford the title product as white fluffy solid.

LC-MS (Method 5): Rt 5.36 min, m/z 743 [M+H]⁺.

Example 10

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-hydroxy-2-(4- hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyI}- phenylcarbamoyl)-ethyl]-methyl-amino}-bicyclo[2.2.1 ]hept-7-yl ester formate

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid (1 S,2R,4S,7S)-7-{[2-(4-formyl-phenylcarbamoyl)-ethyl]-methyl-amino}- bicyclo[2.2.1]hept-2-yl ester (Intermediate 54) and 7-((R)-2-amino-1-hydroxy-ethyl)-4- hydroxy-3H-benzothiazol-2-one hydrochloride by a similar method to that employed for Intermediate 52. The crude product was purified by preparative reversed-phase HPLC (System 1), eluting with 10-25% MeCN/water (0.1% formic acid) over 20 min. The pure fractions were combined and lyophilized to afford the title product as white fluffy solid.

LC-MS (Method 5): Rt 5.69 min, m/z 749 [M+H]⁺.

Example 11

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-hydroxy-2-(8- hydroxy-2-oxo-1,2-dihydro-quinolin-5-yl)-ethylamino]-methyl}- phenylcarbamoyl)-ethyl]-methyl-amino}-bicyclo[2.2.1 ]hept-7-yl ester formate

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid

(i S,2S,4R,7R)-2-{[2-(4-{[(R)-2-(tert-butyl-dimethyl-silanyloxy)-2-(8-hydroxy-2-oxo-1 ,2- dihydro-quinolin-5-yl)-ethylamino]-methyl}-phenylcarbamoyl)-ethyl]-methyl-amino}- bicyclo[2.2.1]hept-7-yl ester (Intermediate 55) by a similar method to that employed for Example 2. The crude product was purified preparative reversed-phase HPLC (System 1), eluting with 10-25% MeCN/water (0.1% formic acid) over 15 min. The pure fractions were combined and lyophilized to afford the title product as white fluffy solid.

LC-MS (Method 5): Rt 5.58 min, m/z 743 [M+H]⁺.

Example 12

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(3-{2-[(R)-2-hydroxy-2- (4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-ethyl}- benzoylamino)-ethyl]-methyl-amino}-bicyclo[2.2.1 ]hept-7-yl ester formate

The title compound was prepared from hydroxy-di-thiophen-2-yl-acetic acid (1S,2S,4R,7R)-2-({2-[3-(2-hydroxy-ethyl)-benzoylamino]-ethyl}-methyl-amino)- bicyclo[2.2.1]hept-7-yl ester (Intermediate 59) by a similar method to that employed for Example 8. The crude product was purified by preparative reversed-phase HPLC (System 1), eluting with 10-25% MeCN/water (0.1% formic acid) over 15 min. The pure fractions were combined and lyophilized to afford the title product as white fluffy solid.

LC-MS (Method 5): Rt 5.67 min, m/z 763 [M+H]⁺.

BIOLOGICAL ASSAYS

The inhibitory effects of compounds of the present invention at the M₃ muscarinic receptor and the β₂ adrenergic receptor may be determined by the following binding assays:

Muscarinic 3 receptor binding assays (plCsn and spot test activity determinations)

The activity of compounds at the M₃ receptor is determined by competition binding of [³H]Λ/-methyl scopolamine (NMS) to CHO-K1 (Chinese Hamster Ovary) cell membranes expressing the human muscarinic acetylcholine M₃ receptor (M₃-ACh) in a scintillation proximity assay (SPA) format.

SPA beads are precoated with membranes and then incubated at 2 mg of beads per well with compounds of the invention, [³H]NMS at 0.2 nM, half Kd (experimentally determined dissociation constant) and assay buffer (20 mM HEPES pH 7.4 containing 5 mM MgCI₂). The assay is conducted in a final volume of 200 μl_ in the presence of 1% (v/v) DMSO. Total binding of [³H]NMS is determined in the absence of competing compound and non-specific binding of [³H]NMS is determined in the presence of 1 μM atropine. The plates are incubated for 16 h at RT and then read on Wallac Microbeta^™ using a normalised ³H protocol.

For plC₅₀ determinations the assay is conducted using serial dilutions of compounds of the invention. The plC₅₀ is defined as the negative logarithm of the concentration of compound required for 50% reduction in specific [³H]-NMS binding. For spot test activity determinations compounds of the invention are incubated at a single 10 nM concentration using the protocol described above. The percentage inhibition of specific [³H]NMS binding is reported.

Compounds 1-12 were tested for binding to the M₃ receptor and the following results obtained:

^* = M₃ binding plC₅₀ reported

β- Adrenergic Receptor Radioligand Binding Assay

Radioligand binding studies utilising [¹²⁵l]-lodocyanopindolol and commercially available cell membranes expressing the human β₂ adrenergic receptor are used to assess the affinity of antagonists for β₂-adrenergic receptor. Membranes and SPA- beads are incubated with [¹²⁵l]-lodocyanopindolol and β₂ antagonist at various concentrations for 3 h at RT in TRIS buffer. The assay is performed in 96-well plates, which are read using the Wallac Microbeta counter.

Where tested, Compounds 1-12 were tested in the β₂ binding assay and were found to have a β₂ plC₅₀ of >5.

Assay for adrenergic β? mediated cAMP production Cell preparation

H292 cells are grown in 225 cm² flasks incubator at 37 ⁰C, 5% CO₂ in RPMI medium containing 10 % (v/v) FBS (foetal bovine serum) and 2 mM L-glutamine. Experimental Method

Adherent H292 cells were removed from tissue culture flasks by treatment with Accutase™ cell detachment solution for 15 min. Flasks are incubated for 15 min in a humidified incubator at 37 ⁰C, 5% CO₂. Detached cells are re-suspended in RPMI media (containing 10% (v/v) FBS and 2 mM L-glutamine) at 0.05 x 10⁶ cells per ml_. 5000 cells in 100 μl_ are added to each well of a tissue-culture-treated 96-well plate and the cells incubated in a humidified incubator at 37 °C, 5% CO₂ overnight. The culture media is removed and cells are washed twice with 100 μL assay buffer and replaced with 50 μL assay buffer (HBSS solution containing 10 mM HEPES Ph 7.4 and 5 mM glucose). Cells are rested at RT for 20 min after which time 25 μL of Rolipram (1.2 mM made up in assay buffer containing 2.4% (v/v) DMSO) is added. Cells are incubated with Rolipram for 10 minutes after which time test compounds are added and the cells are incubated for 60 min at RT. The final Rolipram concentration in the assay is 300 μM and final vehicle concentration is 1.6% (v/v) DMSO. The reaction is stopped by removing supernatants, washing once with 100 μL assay buffer and replacing with 50 μL lysis buffer. The cell monolayer is frozen at -80 °C for 30 min (or overnight).

AlphaScreen™ cAMP detection

The concentration of cAMP (cyclic adenosine monophosphate) in the cell lysate is determined using AlphaScreen™ methodology. The frozen cell plate is thawed for 20 minutes on a plate shaker, then 10 μL of the cell lysate is transferred to a 96-well white plate. 40 μL of mixed AlphaScreen™ detection beads pre-incubated with biotinylated cAMP, is added to each well and the plate incubated at RT for 10 h in the dark. The AlphaScreen™ signal is measured using an EnVision spectrophotometer (Perkin-Elmer Inc.) with the recommended manufacturer's settings. cAMP concentrations are determined by reference to a calibration curve determined in the same experiment using standard cAMP concentrations. Concentration response curves for agonists are constructed and data is fitted to a four parameter logistic equation to determine both the pEC₅₀ and Intrinsic Activity. Intrinsic Activity is expressed as a fraction relative to the maximum activity determined for Formoterol in each experiment.

The compounds of Examples 1-12 were tested in the M₃ receptor binding and β₂ cAMP functional assays and the following results obtained:

^* Intrinsic activity relative to Formoterol is shown in parenthesis. *^* % Inhibition (1O nM). §= β₂ PlC₅₀ data.

BIOLOGICAL EXAMPLES

The inhibitory effects of compounds of the present invention at the M₃ muscarinic receptor and the β₂ adrenergic receptor, may be determined by the following binding assays:

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) are used to assess the affinity of muscarinic antagonists for M2 and M3 receptors. Membranes in TRIS buffer are incubated in 96-well plates with [³H]-NMS and M3 antagonist at various concentrations for 3 hours. Membranes and bound radioligand are then harvested by filtration and allowed to dry overnight. Scintillation fluid is then added and the bound radioligand counted using a Canberra Packard Topcount scintillation counter

The half-life of antagonists at each muscarinic receptor are measured using the alternative radioligand [³H]-QNB and an adaptation of the above affinity assay.

Antagonists are 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 is added to a concentration 25-fold higher than its Kd for the receptor being studied and the incubation continues for various time periods from 15 minutes up to 180 minutes. Membranes and bound radioligand are then harvested by filtration and allowed to dry overnight. Scintillation fluid is 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, ie. to the half life of the antagonists on the receptors.

β- Adrenergic Receptor Radioligand Binding Assay

Radioligand binding studies utilising [¹²⁵l]-lodocyanopindolol and commercially available cell membranes expressing the human β₂ adrenergic receptor are used to assess the affinity of antagonists for β₂-adrenergic receptor. Membranes and SPA- beads are incubated with [¹²⁵l]-lodocyanopindolol and β₂ antagonist at various concentrations for 3 hours at ambient temperature in TRIS buffer. The assay is performed in 96-well plates which are read using the Wallac Microbeta counter.

Analysis of Inhibition of M3 Receptor Activation via Calcium Mobilization In an alternative M3 receptor binding assay, CHO cells expressing the human M3 receptor are 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) is prepared in HBSS buffer with the addition of 5 mM probenecid (pH 7.4). An equal volume of the dye solution (75 μL) is 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 is read on a FLEXstation™ (excitation 488 nm, emission 525 nm) for 15 seconds to determine baseline fluorescence. The muscarinic agonist Carbachol is then added at an EC₈₀ concentration and the fluorescence measured for a further 60 seconds. The signal is 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 is then calculated in order to generate IC₅₀ curves.

The inhibitory effects of compounds of the present invention at the M3 muscarinic Receptor may be evaluated in the following ex-viva and in vivo assays:

Evaluation of potency and duration of action in Isolated Guinea Pig Trachea

Experiments are carried out at 37⁰C in modified Krebs-Henseleit solution, (114 mM NaCI, 15 mM NaHCO₃, 1 mM MgSO₄, 1.3 mM CaCI₂, 4.7 mM KCI, 11.5 mM glucose and 1.2 mM KH₂PO₄ , pH 7.4) gassed with 95 % O₂/5 % CO₂. lndomethacin is added to a final concentration of 3 μM

Tracheae are removed from adult male Dunkin Hartley Guinea pigs and dissected free of adherent tissue before being cut open longitudinally in a line opposite the muscle. Individual strips of 2-3 cartilage rings in width are cut and suspended using cotton thread in 10 ml water-jacketed organ baths and attached to a force transducer ensuring that the tissue is located between two platinum electrodes. Responses are recorded via a MPIOOW/Ackowledge data acquisition system connected to a PC. Tissues are equilibrated for one hour under a resting tone of 1 g and are then subjected to electrical field stimulation at a frequency of 80 Hz with a pulse width of 0.1 ms, a unipolar pulse, triggered every 2 minutes. A "voltage-response" curve is generated for each tissue and a submaximal voltage then applied to every piece of tissue according to its own response to voltage. Tissues are washed with Krebs solution and allowed to stabilize under stimulation prior to addition of test compound. Concentration response curves are obtained by a cumulative addition of test compound in half-log increments. Once the response to each addition had reached a plateau the next addition is made. Percentage inhibition of EFS-stimulated contraction is calculated for each concentration of each compound added and dose response curves constructed using Graphpad Prism software and the EC₅₀ calculated for each compound.

Onset time and duration of action studies are performed by adding the previously determined EC₅₀ concentration of compound to EFS contracted tissues and the response allowed to plateau. The time taken to reach 50 % of this response is determined to be the onset time. Tissues are then washed free of compound by flushing the tissue bath with fresh Krebs solution and the time taken for the contraction in response to EFS to return to 50 % of the response in the presence of compound is measured. This is termed the duration of action.

Methacholine Induced Bronchoconstriction in vivo Male Guinea pigs (Dunkin Hartley), weighing 500-600 g housed in groups of 5 are individually identified. Animals are allowed to acclimatize to their local surroundings for at least 5 days. Throughout this time and study time animals are allowed access to water and food ad libitum.

Guinea pigs are anaesthetized with the inhaled anaesthetic Halothane (5 %). Test compound or vehicle (0.25 - 0.50 mUkg) is administered intranasally. Animals are placed on a heated pad and allowed to recover before being returned to their home cages.

Up to 24 hrs post dosing guinea pigs are terminally anaesthetized with Urethane (250 μg/mL, 2 ml_/kg). At the point of surgical anaesthesia, the jugular vein is cannulated with a portex i.v. cannula filled with heparinised phosphate buffered saline (hPBS) (10

U/mL) for i.v. administration of methacholine. The trachea is exposed and cannulated with a rigid portex cannula and the oesophagus cannulated transorally with a flexible portex infant feeding tube.

The spontaneously breathing animal is then connected to a pulmonary measurement system (EMMS, Hants, UK) consisting of a flow pneumotach and a pressure transducer. The tracheal cannula is attached to a pneumotach and the oesophageal cannula attached to a pressure transducer.

The oesophageal cannula is positioned to give a baseline resistance of between 0.1 and 0.2 cmH20/mLJs. A 2 minute baseline reading is recorded before i.v. administration of methacholine (up to 30 μg/kg, 0.5 mlJkg). A 2 minute recording of the induced constriction is taken from the point of i.v. administration.

The software calculated a peak resistance and a resistance area under the curve

(AUC) during each 2 minute recording period which is used to analyse the bronchoprotective effects of test compounds.

Claims

CLAIMS:

1. A compound of formula (I)

wherein

X is OR⁴ or NR⁵R¹²;

(i) R¹ is d-Cβ-alkyl, (C₃-C₆)cycloalkyl or hydrogen; and R² is a group-Z-NR⁹R¹⁰; and R³ is a lone pair or C₁-CVaIkVl₁ 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 , -Z-NR⁹R¹⁰; and R³ is a lone pair or d-CValkyl, in which cases the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge;

R⁴ is selected from one of the groups of formula (a), (b), (c) or (d):

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

R⁵ is selected from one of the groups of formula (a), (b) or (c):

(a) (b) (C)

R⁶ is CrCβ-alkyl, (C₃-C₆)cycloalkyl or a hydrogen atom;

R and R are a CrCβ-alkyl, (C₃-C₆)cycloalkyl group or halogen;

n and m are independently 0, 1 , 2 or 3;

each occurrence of R^8a and R^8b is independently selected from the group consisting of aryl, aryl-fused-heterocycloalkyl, heteroaryl, C_rC₆-alkyl, (C₃-C₆)cycloalkyl;

each occurrence of R^8c is independently selected from -OH, CrCβ-alkyl, (C₃- C₆)cycloalkyl, hydroxy-CrCe-alkyl, nitrile, a group CONR^8d ₂ and a hydrogen atom;

R^8d is CrC₆-alkyl, (C₃-C₆)cycloalkyl or a hydrogen atom;

R⁹ is a hydrogen atom, (C₃-C₆)cycloalkyl or C_rC₆-alkyl;

R is an aryKCVCValkyl)-, or

group, wherein both CVCValkyl species are optionally substituted by hydroxy;

R is CrC₆-alkyl, (C₃-C₆)cycloalkyl or a hydrogen atom;

Ar¹ is aryl, heteroaryl or (C₃-C₆)cycloalkyl;

Ar² are, independently, aryl, heteroaryl or (C₃-C₆)cycloalkyl; and

Q is an oxygen atom, -CH₂-, -CH₂CH₂- or a bond;

Z is selected from:

(i) a CrC^-alkylene, C₂-C₁₆-alkenylene or C₂-C₁₆-alkynylene group;

or (N) a linker ^*-C(L³)(L⁴)-L-C(L¹)(L²)-^** , wherein:

^* denotes the point of attachment to the non-aromatic nitrogen bearing R¹ and R³ or to the heterocycloalkyl ring formed by R¹ and R²; and ^** denotes the point of attachment to the group NR⁹R¹⁰; L is a hydrocarbyl chain of up to 14 carbon atoms, wherein one, two or three carbon atoms of the chain are replaced by groups independently selected from O, NR⁴⁵, S₁ S(O), S(O)₂, C(O)O, OC(O), NR⁴⁶C(O), C(O)NR⁴⁷, NR⁴⁸S(O)₂, S(O)₂NR⁴⁹, NR⁵⁰C(O)NR⁵¹, NR⁵²S(O)₂NR⁵³,

OC(O)NR⁵⁴, NR⁵⁵C(O)O, provided that any heteroatoms in the chain are separated by at least 2 carbon atoms; and/or wherein up to four carbon atoms of the chain form part of a mono- or bicyclic aliphatic, heteroaliphatic, aromatic or heteroaromatic ring having up to three heteroatoms independently selected from N, O or S, said ring comprising up to 10 ring atoms, and wherein the ring is optionally substituted by up to three substituents independently selected from halogen, S(O)₀.₂R⁵⁶, NR⁵⁷R⁵⁸, S(O)₂NR⁵⁹R⁶⁰, C(O)NR⁶¹R⁶², C(O)OR⁶³, NR⁶⁴S(O)₂R⁶⁵, NR⁶⁶C(O)R⁶⁷, NR⁶⁸C(O)OR⁶⁹, NR⁷⁰C(O)NR⁷¹R⁷², OR⁷³, C_1-6 alkyl and C₃-₆cycloalkyl, and wherein C₁^ alkyl and C_3-6cycloalkyl may be optionally substituted by up to three substituents independently selected from halogen, hydroxyl and C₁^ alkoxy; and the chain may comprise up to three of such rings each selected independently; wherein R⁵⁶, R⁶⁵ and R⁶⁹ each independently represent C_1-6 alkyl or C₃. ₆cycloalkyl, wherein C₁^ alkyl and C_3-6 cycloalkyl may be optionally substituted by up to three substituents independently selected from halogen, hydroxyl, C₁- ₆ alkoxy; and R⁴⁵, R⁴⁶, R⁴⁷, R⁴⁸, R⁴⁹, R⁵⁰, R⁵¹, R⁵², R⁵³, R⁵⁴, R⁵⁵, R⁵⁷, R⁵⁸, R⁵⁹, R⁶⁰,

R⁶¹, R⁶², R⁶³, R⁶⁴, R⁶⁶, R⁶⁷, R⁶⁸ , R⁷⁰, R⁷¹, R⁷² and R⁷³ each independently represent hydrogen, or C_1-6 alkyl or C₃.₆ cycloalkyl, wherein C^₆ alkyl and C_3-6 cycloalkyl may be optionally substituted by up to three substituents independently selected from halogen, hydroxyl, C_1-6 alkoxy; or any of R⁵⁷ and R⁵⁸, R⁵⁹ and R⁶⁰, R⁶¹ and R⁶² or R⁷¹ and R⁷², together with the nitrogen atom to which they are both attached, may form a 4 to 8 membered aliphatic heterocyclic ring, wherein the aliphatic heterocyclic ring may comprise up to three heteroatoms independently selected from N₁O and S, wherein the ring may be optionally substituted by up to three substituents independently selected from halogen, hydroxyl and C₁^ alkyl or C₃.₆cycloalkyl, wherein C₁^ alkyl and C_3-6cycloalkyl may be optionally substituted by one or more substituents independently selected from halogen and hydroxyl; and wherein the chain may additionally comprise up to three carbon- carbon double bonds; wherein the chain may additionally comprise up to three carbon- carbon triple bonds; L¹ and L² each independently represent hydrogen, C^₆ alkyl or C_3-6 cycloalkyl;

L³ and L⁴ each independently represent hydrogen, C_1-6 alkyl or C₃.₆ cycloalkyl, wherein C₁^ alkyl and C_3-6 cycloalkyl may be optionally substituted by one or more substituents independently selected from halogen and hydroxyl; in addition L¹ and/or L³ may be linked to carbon atoms of the hydrocarbyl chain in linker L to form aliphatic rings of up to 6 ring atoms, wherein each ring may comprise up to three heteroatoms independently selected from N, O and S; with the proviso that when X is OR4 then Z is a linker ^*-C(L³)(L⁴)-L-C(L¹)(L²)-^**;

or a pharmaceutically acceptable salt thereof.

2. A compound as claimed in claim 1 , or a pharmaceutically acceptable salt thereof, wherein

R¹ is d-Ce-alkyl; R² is a group -Z-NR⁹R¹⁰ and R³ is a lone pair or d-C₆-alkyl in which cases the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge, or:

R¹ and R² together with the nitrogen to which they are attached represent a heterocycloalkyl ring, said ring being substituted by a group -Z-NR⁹R¹⁰ and R³ is a lone pair or C_rC₆-alkyl, in which cases the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge; or: R¹ and R³ together with the nitrogen to which they are attached represent a heterocycloalkyl ring, and R² is a group -Z-NR⁹R¹⁰, in which cases the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge;

R⁴ is selected from one of the groups of formula (a), (b) or (c):

(a) (b) (C) wherein Ar¹and Ar² are as defined in claim 1 ;

R⁶ is a hydrogen atom;

R and R are independently a d-Cβ-alkyl group or halogen;

n and m are independently 0, 1 , 2 or 3;

R^8a and R^8b are independently selected from the group consisting of aryl, heteroaryl, CrCβ-alkyl, cycloalkyl;

R is -OH, Ci-C₆-alkyl, hydroxy-d-Ce-alkyl, or a hydrogen atom;

R⁹ is a hydrogen atom;

R¹⁰ is a group ArCH(OH)CH₂- Wherein Ar is a group

wherein M¹ is S, C(O), NA⁵, CA⁶A⁷, CH₂CH₂, CH=CH, CH₂O or OCH₂;

Mf is S, C(O), NA*, CA⁶A', CH₂CH₂, CH=CH, CH₂O or OCH₂;

A¹, A², A³ and A⁴ are, independently, hydrogen, halogen, trifluoromethyl, cyano, carboxy, hydroxy, nitro, S(O)₂A⁸, NA⁹S(O)₂A¹⁰, C(O)NA¹¹A¹², NA¹³C(O)A¹⁴, C_1-6 alkyl,

C_1-6 alkoxy, C(O)(C₁^ alkyl) or C(O)OC_1-6 alkyl; and A³ can also be CH₂OH, NHCHO, NHS(O)₂NA¹⁵A¹⁶ or NHSO₂A¹⁷;

A⁵, A⁶, A⁷, A⁹, A¹¹, A¹², A¹³, A¹⁴, A¹⁵ or A¹⁶ are, independently, hydrogen or C_1-6 alkyl; A⁸, A¹⁰ and A¹⁷ are, independently, C_1-6 alkyl.

3. A compound as claimed in claim 2, or a pharmaceutically acceptable salt thereof, where Ar is selected from:

wherein

M¹ is S, CH=CH, CH₂O or OCH₂;

M² is S, CH=CH, CH₂O or OCH₂; A¹, A², and A⁴ are, independently, hydrogen, halogen, C_1-6 alkyl, C_1-6 alkoxy; and A³ can also be CH₂OH, NHCHO, NHS(O)₂NA¹⁵A¹⁶ or NHSO₂A¹⁷;

A or A are independently selected from hydrogen or C_1-6 alkyl;

A¹' is C_1-6 alkyl.

4. A compound as claimed in claim 3, or a pharmaceutically acceptable salt thereof, wherein Ar is selected from the group

5. A compound according to claim 1 , selected from:

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[3-(4-{[(R)-2-hydroxy- 2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}-phenoxy)- propyl]-methyl-amino}-bicyclo[2.2.1 ]hept-7-yl ester;

9-Hydroxy-9H-xanthene-9-carboxylic acid (1 S,2S,4R,7R)-2-{[3-(4-{[(R)-2- hydroxy-2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}- phenoxy)-propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester; g-Hydroxy-QH-xanthene-g-carboxylic acid (1S,2S,4R,7R)-2-{[3-(4-{[(R)-2- hydroxy-2-(8-hydroxy-2-oxo-1 ,2-dihydro-quinolin-5-yl)-ethylamino]-methyl}-phenoxy)- propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester; θ-Hydroxy-ΘH-xanthene-θ-carboxylic acid (1 S,2S,4R,7R)-2-{[3-(4-{[(R)-2- hydroxy-2-(5-hydroxy-3-oxo-3,4-dihydro-2H-benzo[1 ,4]oxazin-8-yl)-ethylamino]- methyl}-phenoxy)-propyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester;

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-[3-(4-{[(R)-2-hydroxy-2- (4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}-phenoxy)- propylamino]-bicyclo[2.2.1]hept-7-yl ester;

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-hydroxy- 2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}- benzoylamino)-ethyl]-methyl-amino}-bicyclo[2.2.1 ]hept-7-yl ester; Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-hydroxy-

2-(8-hydroxy-2-oxo-1 ,2-dihydro-quinolin-5-yl)-ethylamino]-methyl}-benzoylamino)- ethyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester; Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-hydroxy- 2-(4-hydroxy-2-oxo-2,3-dihydro-benzothiazol-7-yl)-ethylamino]-methyl}- phenylcarbamoylj-ethyll-methyl-aminoj-bicyclo^^.ijhept^-yl ester;

Hydroxy-di-thiophen-2-yl-acetic acid (1 S,2S,4R,7R)-2-{[2-(4-{[(R)-2-hydroxy- 2-(8-hydroxy-2-oxo-1 ,2-dihydro-quinolin-5-yl)-ethylamino]-methyl}-phenylcarbamoyl)- ethyl]-methyl-amino}-bicyclo[2.2.1]hept-7-yl ester; and

6. A compound or a pharmaceutically acceptable salt thereof as claimed in any of the preceding claims, for use in therapy.

7. A pharmaceutical composition comprising a compound as claimed in any of claims 1 to 5 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

8. A pharmaceutical composition as claimed in claim 7 in a form suitable for inhalation.

9. Use of a compound or a pharmaceutically acceptable salt thereof as claimed in any of claims 1 to 5 for the manufacture of a medicament for use in the treatment of prevention of a disease or condition in which M3 muscarinic receptor activity and β-adrenergic receptor activity is implicated.

10. A method of treatment of a disease or condition in which M3 muscarinic receptor activity and β-adrenergic receptor activity is implicated comprising administration to a subject in need thereof of an effective amount of a compound as claimed in any of claims 1 to 5.

11. The use of a compound of formula (Ia):

(Ia) wherein

R¹ is C_rC₆-alkyl or a hydrogen atom; and R² is a hydrogen atom or a group -R⁵, or a group, -Z-Y-R⁵, or a group -Z-NR⁹R¹⁰, or a group -Z-N(R⁹)C(O)R¹¹; and R³ is a lone pair, or CrC₆-alkyl in which case the nitrogen atom to which it is attached is a quaternary nitrogen and carries a positive charge;

R⁴ is selected from one of the groups of formula (a), (b), (c) or (d);

(a) (b) (C) (d) Z is a CrC^-alkylene, C₂-C₁₆-alkenylene or C₂-C₁₆-alkynylene group;

Y is a bond or oxygen atom;

R⁵ is an C_rC₆-alkyl, aryl, arylalkyl; aryl-fused-cycloalkyl, aryl-fused-heterocycloalkyl, heteroaryl, aryKCVCβ-alkyl)-, heteroaryKCVCβ-alkyl)-, cycloalkyl or heterocycloalkyl group;

R⁶ is C_rC₆-alkyl or a hydrogen atom;

R^7a and R^7b are a d-Ce-alkyl group or halogen;

n and m are independently 0, 1 , 2 or 3;

R^8a and R^8b are independently selected from the group consisting of aryl, aryl-fused- heterocycloalkyl, heteroaryl, CVCe-alkyl, cycloalkyl and hydrogen;

R^8c is -OH, Ci-C₆-alkyl, hydroxy-Ci-C₆-alkyl, or a hydrogen atom;

R⁹ and R¹⁰ are independently a hydrogen atom, CrC₆-alkyl, aryl, aryl-fused- heterocycloalkyl, aryl-fused-cycloalkyl, heteroaryl, aryKd-Cβ-alkyl)-, or heteroaryl(Cr C₆-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; R¹¹ is C_rC₆-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;

12. A method of treatment of a disease or condition in which M3 muscarinic receptor activity and β-adrenergic receptor activity is implicated comprising administration to a subject in need thereof of an effective amount of a compound of formula (Ia), as defined in claim 11.

13. The use according to claim 9 or claim 11 , or the method according to claim 10 or claim 12, wherein the disease or condition is a respiratory-tract disorder.

14. The use according to claim 9 or claim 11 , or the method according to claim 10 or claim 12, wherein the disease or condition is chronic obstructive lung disease, chronic bronchitis, asthma, chronic respiratory obstruction, bronchial hyperactivity, pulmonary fibrosis, pulmonary emphysema, or allergic rhinitis.

15. Use of a compound or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1 to 5 in combination with (i) an inhaled corticosteroid such as fluticasone propionate, ciclesonide, mometasone furoate or budesonide and/or (ii) an inhaled PDE4 inhibitor, such as roflumilast, cilomilast or tofimilast.