COMPOUNDS AND THEIR USE

Field of the Invention

This invention relates to heterocyclic compounds, pharmaceutical compositions containing them, methods for their preparation, their use in the treatment of diseases where enhanced M3 receptor activation is implicated and their Use in the treatment of diseases where compounds possessing both muscarinic receptor antagonist and β2- agonist activity present in the same molecule (bifunctional molecules) are useful (such as in the treatment of asthma or COPD).

Background to the invention

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

There are five subtypes of muscarinic acetylcholine receptors (mAChRs), termed M1-M5, and each is the product of a distinct gene and each displays unique pharmacological properties. mAChRs are widely distributed in vertebrate organs, and these receptors can mediate both inhibitory and excitatory actions. For example, in smooth muscle found in the airways, bladder and gastrointestinal tract, M3 mAChRs mediate contractile responses (reviewed by Caulfield, 1993, Pharmac. 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 Med., 158 (5 part 3) S 154 - 160). M3 receptors on airway smooth muscle mediate contraction and therefore bronchoconstriction. Stimulation of M3 receptors localised to submucosal glands results in mucus secretion.

Increased signalling through muscarinic acetylcholine receptors has been noted in a variety of different pathophysiological states including asthma and COPD. In COPD, vagal tone may either be increased (Gross 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 M2 receptor activity which results in increased levels of acetylcholine release following vagal nerve stimulation (Fryer et al, 1999, Life Sci., 64, (6-7) 449-455). The resultant increased activation of M3 receptors leads to enhanced airway obstruction. Thus the identification of potent muscarinic receptor antagonists would be useful for the therapeutic treatment of those disease states where enhanced M3 receptor activity is implicated. Indeed, contemporary treatment strategies currently support regular use of M3 antagonist bronchodilators as first-line therapy for COPD patients (Pauwels etal. 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.

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

Ipratropium

Chem. Pharm. Bull. 27(12) 3149-3152 (1979) and J. Pharm. Sci 69 (5) 534-537

(1980) describe f uryl derivatives as possessing atropine-like activities.

Med. Chem. Res 10 (9), 615-633 (2001) describes isoxazoles and δ ² -isoxazolines as muscarinic antagonists.

WO97/30994 describes oxadiazoles and thiadiazoles as muscarinic receptor antagonists.

EP0323864 describes oxadiazoles linked to a mono- or bicyclic ring as muscarinic receptor modulators.

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 or other muscarinic antagonist; WO02/060532 describes a combination of ipratropium with a β2-agonist; and, WO02/060533 describes a combination of oxotropium with a β2-agonist. 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 another active ingredient, for example a steroid. Such bifunctional 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 an M3 antagonist to a β2-agonist.

Summary of.the Invention

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

wherein (i) R ¹ is d-C _ff -alkyl; and R ² is a group -(Z) _p -R ⁷ , -Z-Y-R ⁷ , -Z-NR ⁹ R ¹⁰ , -Z-CO-NR ⁹ R ¹⁰ , -L ^a -Z ¹ or -Z-C(O)-R ⁷ ; and R ³ is a lone pair or R ³ is C _r C ₆ -alkyl, in which case the nitrogen to which they are attached is quaternary and carries a positive charge; PROVIDED THAT R ¹ , R ² and R ³ do not all represent CH ₃ and that when R ³ is a lone

pair then R ¹ and R ² do not both represent CH ₃ ; or

(ii) R ¹ and R ³ together with the nitrogen to which they are attached form a heterocycloalkyl ring, and R ² is a group -(Z) _p -R ⁷ , -Z-Y-R ⁷ , -Z-NR ⁹ R ¹⁰ , -Z-CO-NR ⁹ R ¹⁰ , -L ^a -Z ¹ or -Z-C(O)-R ⁷ , in which case the nitrogen to which they are attached is quaternary and carries a positive charge; or

(iii) R ¹ and R ² together with the nitrogen to which they are attached form a heterocycloalkyl ring, said ring being substituted by a group -R ⁷ , -Y-R ⁷ , -Z-Y-R ⁷ , — Z- NR ⁹ R ¹⁰ , -Z-CO-NR ⁹ R ¹⁰ , -L ^a -Z ¹ or -Z-C(O)-R ⁷ ; and R ³ is a lone pair or R ³ is C _r C ₆ -alkyl, in which case the nitrogen to which they are attached is quaternary and carries a positive charge;

p is O or 1 ;

R ⁴ and R ⁵ are independently selected from the group consisting of aryl, aryl-fused- heterocycloalkyl, heteroaryl, C _r C ₆ -alkyl, cycloalkyl;

or R ⁴ and R ⁵ are joined together to form a tricyclic ring so that the group R ⁴ R ⁵ R ⁶ C-

represents the group , where R ^6a is -OH, C _r C ₆ -alkyl or a hydrogen atom, and Q is an oxygen atom, -CH ₂ -, -CH ₂ CH ₂ - or a bond;

R ⁶ is -OH, C _r C ₆ -alkyl, C _r C ₆ -alkoxy, hydroxy-CrC ₆ -alkyl, nitrile, a group CONR ¹³ ₂ or a hydrogen atom;

one of W, V and A is N or NR ¹¹ ; another of W, V and A is N, O, S or CR ⁸ ; and the last one of W ₁ V and A is N or CR ⁸ ; PROVIDED THAT when A is an oxygen or sulfur atom and W is a nitrogen atom, then V is not a group CR ⁸ ;

X is a CrC^-alkylene, C ₂ -C ₁₂ -alkenylene or C ₂ -C ₁₂ -alkynylene group;

R ⁷ is an C _r C ₆ -alkyl, aryl, aryl-fused-cycloalkyl, aryl-fused-heterocycloalkyl, heteroaryl,

aryl(C _r C ₈ -alkyl)-, heteroaryl(C ₁ -C ₈ -alkyl)-, cycloalkyl or heterocycloalkyl group;

Z is a Ci-C ₁₆ -alkylene, C ₂ -Ci ₆ -alkenylene or C ₂ -Ci ₆ -alkynylene group;

Y is an oxygen atom, a group -S(O) _n , C(O)O, OC(O), N(R ¹² )S(O) ₂ or S(O) ₂ N(R ¹² );

n is 0, 1 or 2;

R ⁹ and R ¹⁰ are independently a hydrogen atom, CrC ₆ -alkyl, aryl, aryl-fused- heterocycloalkyl, aryl-fused-cycloalkyl, heteroaryl, aryl(Ci-C ₆ -alkyl)-, or heteroary^Cr C ₆ -alkyl)- group; or R ⁹ and R ¹⁰ 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;

R ⁸ , R ¹¹ , R ¹² and R ¹³ are, independently, hydrogen atom or C _r C ₆ -alkyl group;

L ^a is a divalent linker radical of formula (Ic):

wherein L represents a linker comprising a hydrocarbyl chain of up to 14 carbon atoms, wherein the chain may additionally comprise up to three carbon-carbon double bonds, and, wherein the chain may additionally comprise up to three carbon-carbon triple bonds;

L ¹ and L ² each independently represent hydrogen, Ci- ₆ alkyl or C _3-6 cycloalkyl;

L ³ and L ⁴ each independently represent hydrogen, Ci- ₆ alkyl or C _3-6 cycloalkyl, wherein C ₁ ^ alkyl and C _3-6 cycloalkyl maybe optionally substituted by one or more substituents independently selected from halogen and hydroxyl; and * denotes the point of attachement of the group of formula (I) to the non- aromatic nitrogen bearing R ¹ and R ³ , and ** denotes the point of attachment to the group Z ¹ ; and

Z ¹ is a moiety having β2- adrenoreceptor agonist activity;

wherein, unless otherwise specified, each occurrence of alkyl, heterocycloalkyl, aryl, aryl-fused-heterocycloalkyl, heteroaryl, cycloalkyl, alkoxy, alkylene, alkenylene, alkynylene or aryl-fused-cycloalkyl may be optionally substituted; and

wherein each alkenylene chain may contain 1 , 2 or 3 carbon-carbon double bonds and each alkynylene chain may contain 1 , 2 or 3 carbon-carbon triple bonds;

or a pharmaceutically acceptable salt thereof.

The present invention also encompasses a subset of the compounds of formula (I), selected from compounds of formula (Ia):

wherein

(i) R ¹ is d-Cβ-alkyl; and R ² is a group -(Z) _p -R ⁷ , -Z-Y-R ⁷ , -Z-NR ⁹ R ¹⁰ , -Z-CO-NR ⁹ R ¹⁰ or -Z-C(O)-R ⁷ ; and R ³ is C _r C ₆ -alkyl; PROVIDED THAT R ¹ , R ² and R ³ do not all represent CH ₃ and that when R ³ is a lone pair then R ¹ and R ² do not both represent CH ₃ ; or

(ii) R ¹ and R ³ together with the nitrogen to which they are attached form a heterocycloalkyl ring, and R ² is a group -(Z) _p -R ⁷ , -Z-Y-R ⁷ , -Z-NR ⁹ R ¹⁰ , -Z-CO-NR ⁹ R ¹⁰ or -Z-C(O)-R ⁷ ; or

(iii) R ¹ and R ² together with the nitrogen to which they are attached form a heterocycloalkyl ring, said ring being substituted by a group -R ⁷ , -Y-R ⁷ , -Z-Y-R ⁷ , -Z- NR ⁹ R ¹⁰ , -Z-CO-NR ⁹ R ¹⁰ or -Z-C(O)-R ⁷ ; and R ³ is C _r C ₆ -alkyl;

p, R ⁴ , R ⁵ , R ⁶ , W, V ₁ A, X, R ⁷ , Z, Y, R ⁹ , R ¹⁰ , alkyl and heterocycloalkyl are as defined above; and

D ^" is a pharmaceutically acceptable counter-ion.

The present invention also encompasses a further subset of the compounds of formula (I), selected from compounds of formula (Ib):

wherein

(i) R ¹ is C _r C ₆ -alkyl; and R ³ is lone pair or C _r C ₆ -alkyl; or

(ii) R ¹ and R ³ together with the nitrogen to which they are attached form a heterocycloalkyl ring;

R ⁴ , R ⁵ , R ⁶ , W, V, A, X, L ^a , Z ¹ , alkyl and heterocycloalkyl are as defined above;

or a pharmaceutically acceptable salt thereof.

In one aspect, the present invention provides a pharmaceutically acceptable salt of a compound of formula (Ia) as herein defined.

In another aspect, the present invention provides a prodrug of a compound of formula (I), (Ia) or (Ib) 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) ₁ (Ia) or (Ib) 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), (Ia) or (Ib) as herein defined, or an N-oxide, prodrug or pharmaceutically acceptable salt thereof

In one aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein each alkyl, heterocycloalkyl, aryl, aryl-fused-heterocycloalkyl, heteroaryl, cycloalkyl, alkoxy, alkylene, alkenylene, alkynylene or aryl-fused-cyeloalkyl group is unsubstituted.

In one aspect, the present invention provides a compound of formula (I) or (Ia) wherein R ¹ , R ² and R ³ are not all C ₁ -C ₆ alkyl. In another aspect, the present invention provides a compound of formula (I) or (Ia) wherein R ¹ , R ² and R ³ are not all unsubstituted C ₁ -C ₆ alkyl.

In another aspect, the present invention provides a compound of formula (I) or (Ia) wherein R ¹ and R ³ are, independently, C _r C ₆ -alkyl; and R ² is a group -Z-Y-R ⁷ ; and Y,

Z and R ⁷ are as defined herein.

• In yet another aspect, the present invention provides a compound of formula (I) or (Ia) wherein R ¹ and R ³ are, independently, C _r C ₆ -alkyl; and R ² is a group -(Z) _p -R ⁷ ; and Z, p and R ⁷ are as defined herein, PROVIDED THAT R ¹ , R ² and R ³ do not all represent

CH ₃ .

In a further aspect, the present invention provides a compound of formula (I) or (Ia) wherein R ² is a group -(Z) _p -R ⁷ ; and Z, p and R ⁷ are

In one aspect, the present invention provides a compound of formula (I) or (Ia) wherein Y is O.

In one aspect, the present invention provides a compound of formula (I) or (Ia) wherein Z is optionally substituted C ₁ -C ₆ alkylene, optionally substituted C ₂ -C ₆ - alkenylene, or optionally substituted C ₂ -C ₆ -alkynylene group. In a further aspect, the present invention provides a compound of formula (I) or (Ia) wherein Z is optionally substituted C ₁ -C ₆ alkylene.

In one aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein X is optionally substituted C ₁ -C ₆ alkylene. In another aspect of the invention X is C ₁ -C ₂ alkylene.

In one aspect, the present invention provides a compound of formula (I) or (Ia) wherein R ⁷ is an aryl (for example phenyl), aryl-fused-cycloalkyl (for example indanyl) or aryl(C _r C ₈ -alkyl)- (for example benzyl or 2-phenyleth-1-yl) group.

In another aspect, the present invention provides a compound of formula (I) or (Ia)

wherein R ⁷ is selected from aryl (for example phenyl), aryl(C _r C ₈ -alkyl)- (for example benzyl or 2-phenyleth-1-yl), aryl-fused-heterocycloalkyl (for example dihydrobenzofuranyl) and heteroaryl (for example isoxazolyl).

In one aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein the 5-membered ring containing W, V and A is selected from:

R -.1'1

_R \ *\ N-S

N-O N-N N-N N-N

4.* JL N ^ N N N

and wherein the bond marked * is attached to the group R ⁴ R ⁵ R ⁶ C-, and the bond marked * ^* is attached to the group -X-; and R ¹¹ is as defined above.

In another aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein the 5-membered ring containing W, V and A is selected from:

wherein the bond marked * is attached to the group R ⁴ R ⁵ R ⁶ C-, and the bond marked * ^* is attached to the group -X-; and R ¹¹ is as defined above.

In a further aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein the 5-membered ring containing W, V and A is selected from:

O-N N-O

^• λ*- ^~ --V-- wherein the bond marked ^* is attached to the group R ⁴ R ⁵ R ⁶ C-, and the bond marked ** is attached to the group -X-; and R ¹¹ is as defined herein.

In a yet further aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein the 5-membered ring containing W, V and A is selected from:

N-N N-O O-N

---"x /~--- ** * ^^\ ^y y~^ _^ . * _{* *} ----^ /~~~~- ** * O N N

wherein the bond marked * is attached to the group R ⁴ R ⁵ R ⁶ C-, and the bond marked ^** is attached to the group -X-; and R ¹¹ is as defined herein.

In one aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein R ¹¹ is hydrogen or C ₁ -C ₃ alkyl.

In one aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein R ⁶ is hydroxy, Ci-C ₄ alkyl (such as methyl), C ₁ -C ₄ alkoxy (such as methoxy) or nitrile. In another aspect, R ⁶ is hydroxy or C ₁ -C ₄ alkyl (such as methyl). In a further aspect, R ⁶ is hydroxy.

In one aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein R ⁴ and R ⁵ are, independently, aryl (such as phenyl), C ₄ -C ₈ cycloalkyl (such as cyclopentyl or cyclohexyl) or heteroaryl (such as thienyl).

In another aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein R ⁴ is aryl (such as phenyl) or heteroaryl (such as thienyl).

In yet another aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein R ⁵ is C ₄ -C ₈ cycloalkyl (such as cyclopentyl or cyclohexyl), aryl (such as phenyl) or heteroaryl (such as thienyl).

In a further aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein R ⁴ is aryl (for example phenyl) and R ⁵ is C ₄ -C ₇ cycloalkyl (for example cyclopentyl or cyclohexyl).

In a further aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein R ⁴ and R ⁵ are both aryl (for example phenyl), or R ⁴ and R ⁵ are both heteroaryl (for example thienyl).

In a further aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein R ⁴ is aryl (for example phenyl), R ⁵ is C ₄ -C ₈ cycloalkyl (for example cyclopentyl or cyclohexyl), R ⁶ is -OH and the carbon to which R ⁴ , R ⁵ and R ⁶ are attached has the (R)- absolute configuration.

In one subset of the compounds of formula (Ia):

R ¹ is CrC _β -alkyl; R ² is -(Z) _p -R ⁷ or a group -Z-Y-R ⁷ ; and R ³ is C _r C ₆ -alkyl; PROVIDED THAT R ¹ , R ² and R ³ do not all represent CH ₃ ;

p is 0 or 1 ;

R ⁴ and R ⁵ are independently selected from the group consisting of aryl, heteroaryl, C ₁ - C ₆ -alkyl, cycloalkyl;

R ⁶ is -OH;

W, V and A are as defined herein;

X is an CVC^-alkylene, C ₂ -C ₁₆ -alkenylene or C ₂ -Ci ₆ -alkynylene group;

Z is an CrC^-alkylene, C ₂ -Ci ₆ -alkenylene or C ₂ -Ci ₆ -alkynylene group;

Y is oxygen atom or a group -S(O) _n , C(O)O, OC(O), N(R ¹² )S(O) ₂ or S(O) ₂ N(R ¹² );

n is an integer selected from 0, 1 or 2;

R ⁷ is aryl, heteroaryl, aryl(C _r C ₆ -alkyl) or aryl-fused-heterocycloalkyl; and

D ^" is a pharmaceutically acceptable counter-ion.

Particular combinations of W, V and A in the compounds of formula (I), (Ia) and (Ib) include:

(a) W is a group CR ⁸ , V is an oxygen atom and A is a nitrogen atom;

(b) W is a group CR ⁸ , V is a sulfur atom and A is a nitrogen atom;

(c) W is a group CR ⁸ , V is a nitrogen atom and A is an oxygen atom; (d) W is a group CR ^a , V is a nitrogen atom and A is a sulfur atom;

(e) W is a nitrogen atom V is a nitrogen atom and A is an oxygen atom;

(f) W is a nitrogen atom, V is an oxygen atom and A is a nitrogen atom;

(g) W is an oxygen atom, V is a nitrogen atom and A is a nitrogen atom;, (h) W is a group NR ¹¹ , V is a nitrogen atom and A is a group CR ⁸ ; (i) W is a nitrogen atom, V is an oxygen atom and A is a group CR ⁸ ;

(j) W is an oxygen atom, V is a nitrogen atom and A is a group CR ⁸ ;

(k) W is a nitrogen atom, V is a sulfur atom and A is a nitrogen atom;

(I) W is a nitrogen atom, V is a nitrogen atom and A is a sulfur atom;

(m) W is a sulfur atom, V is a nitrogen atom and A is a nitrogen atom; (n) W is a nitrogen atom, V is a group CR ⁸ and A is a group NR ¹¹ .

In one aspect, the present invention provides a compound of formula (I) or (Ib) wherein Z ¹ is a group of formula (A) or (B):

wherein Ar represents a group selected from the following;

wherein

M ¹ is S, C(O), NA ⁵ , CA ⁶ A ⁷ , CH ₂ CH ₂ , CH=CH, CH ₂ O or OCH ₂ ;

M ² is S, C(O), NA ⁵ , CA ⁶ AS CH ₂ CH ₂ , CH=CH, CH ₂ O or OCH ₂ ;

A ¹ , A ² , A ³ and A ⁴ are, independently, hydrogen, halogen, trifluoromethyl, cyano, carboxy, hydroxy, nitro, S(O) ₂ A ⁸ , NA ⁹ S(O) ₂ A ¹⁰ , C(O)NA ¹¹ A ¹² , NA ¹³ C(O)A ¹⁴ , C _1-6 alkyl, C _1-6 alkoxy, C(O)(C ₁ * alkyl) or C(O)OC ₁ * alkyl;

and A ^d can also be CH ₂ OH, NHCHO, NHS(O) ₂ N A ^1& A ^1b or NHSO ₂ A 0 ¹ 7'.;

A ⁵ , A ⁶ , A ⁷ , A ⁹ , A ¹¹ , A ¹² , A ¹³ , A ¹⁴ , A ¹⁵ or A ¹⁶ are, independently, hydrogen or C _1-6 alkyl; A ⁸ , A ¹⁰ and A ¹⁷ are, independently, Ci_ ₆ alkyl, and;

^*** represents the attachment point of the group of formula (A) or (B) to the group of formula (Ic).

In yet another aspect Z ¹ is a group selected from:

The group of formula (Ic)

Conveniently L represents a linker comprising a hydrocarbyl chain of up to 12 carbon atoms or of up to 10 carbon atoms or of up to 8 carbon atoms.

Conveniently the chain may additionally comprise up to two carbon-carbon double bonds or a single carbon-carbon double bond.

Conveniently the chain may additionally comprise up to two carbon-carbon triple bonds or a single carbon-carbon triple bond.

Conveniently each of L ¹ , L ² , L ³ and L ⁴ represent independently hydrogen or a C ₁ ^ alkyl group.

Conveniently: L ¹ and L ² each independently represent hydrogen, C _1-4 alkyl or C _3-6 cycloalkyl. Currently preferred is where L ¹ and L ² are each hydrogen.

L ³ and L ⁴ each independently represent hydrogen, C _1-4 alkyl or C ₃ . ₆ cycloalkyl, which C _1-4 alkyl and C ₃ ^ cycloalkyl may be optionally substituted by one or more substituents

independently selected from halogen and hydroxyl. Currently preferred is where L ³ and L ⁴ are each hydrogen.

In some compounds of the invention, the radical -C(L ₁ )(La)-L-C(L ₃ )(L ₄ )- is -(CH ₂ )r-, wherein r = 6, 7, 8 or 9.

The groups of formula (A) and (B)

In the group of formula (A) and (B): Conveniently Ar is selected from

wherein

M ¹ is S, CH=CH, CH ₂ O or OCH ₂ ;

M ² is S, CH=CH, CH ₂ O or OCH ₂ ;

A ¹ , A ² , and A ⁴ are, independently, hydrogen, halogen, C _1-6 alkyl, C _1-6 alkoxy; and A ³ can also be CH ₂ OH, NHCHO, NHS(O) ₂ NA ¹⁵ A ¹⁶ or NHSO ₂ A ¹⁷ ;

A ¹⁵ or A ¹⁶ are independently selected from hydrogen or C _1-6 alkyl;

A ¹⁷ is C _1-6 alkyl;

Examples of C ₁ ^ alkyl include C _1-4 alkyl and C _1-2 alkyl. Examples of C _1-6 alkoxy include Ci _-4 alkoxy and C _1-2 alkoxy.

More conveniently Ar is selected from:

wherein A ¹ , A ² and A ⁴ are all hydrogen, A ³ is CH ₂ OH, NHCHO and M ¹ is CH=CH or S.

In the compounds of formula (I), (Ia) and (Ib), as defined and further particularized above, R ^s may be hydrogen.

Furthermore, in the compounds of formula (I), (Ia) and (Ib), as defined and further particularized above, R ⁴ and R ⁵ may both be phenyl and R ⁶ may be -OH, or R ⁴ and R ⁵ may both be thienyl or R ⁴ is phenyl and R ⁵ is cycloalkyl.

It will be appreciated that in the compounds of formula (1), (Ia) and (Ib) the carbon atom to which R ⁴ , R ⁵ and R ⁶ are attached can be an asymmetric centre so compounds of the invention may be in the form of single enantiomers or mixtures of enantiomers.

In a further aspect, the present invention provides a compound of formula (I), (Ia) or (Ib) wherein the carbon to which R ⁴ , R ⁵ and R ⁶ are attached has the (R) absolute configuration.

In one aspect, the present invention provides compounds of formula (1) or (Ia) selected from:

[5-(Hydroxy-diphenyl-methyl)-1 ,3,4-oxadiazol-2-ylmethyl]-dimethyl-(3-phenoxy- propyl)-ammonium; [3-(Hydroxy-diphenyl-methyl)-1 ,2,4-oxadiazol-5-ylmethyl]-dimethyl-(3-phenoxy- propyl)-ammonium;

[5-(Hydroxy-diphenyl-methyl)-1 ,2,4-oxadiazol-3-ylmethyl]-dimethyl-(3-phenoxy- propyl)-ammonium;

[4-(Hydroxy-diphenyl-methyl)-oxazol-2-ylmethyl]-dimethyl- (3-phenoxy-propyl)- ammonium;

,[4-(Hydroxy-diphenyl-methyl)-thiazol-2-ylmethyl]-dimethy l-(3-phenoxy-propyl)-

ammonium;

[5-(Hydroxy-diphenyl-methyl)-oxazol-2-ylmethyl]-dimethyl- (3-phenoxy-propyl)- ammonium;

[5-(Hydroxy-diphenyl-methyl)-thiazol-2-ylmethyl]-dimethyl -(3-phenoxy-propyl)- ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-5-yl methyl]-dimethyl- (3-phenoxyl-propyl)-ammonium;

[5-(CyclohexyI-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-3-yl methyl]-dimethyl- (3-phenoxyl-propyl)-ammonium; [5-(Cyclopentyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-3-yI methyl]-dimethyl-

(3-phenoxyl-propyl)-ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl- (3-phenoxy-propyl)-ammonium;

[δ-CCyclopentyl-hydroxy-phenyl-methylJ-fi .S^Joxadiazol^-ylmethylJ-dimethyl- (3-phenoxy-propyl)-ammonium;

[5-(Hydroxy-di-thiophen-2-yl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl-(3- phenoxy-propyl)-ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-thiazol-2-ylmethyl] -dimethyl-(3- phenoxy-propyl)-ammonium; [3-Hydroxy-diphenyl-methyl)-isoxazol-5-ylmethyl]-dimethyl-(3 - phenoxypropyl)ammonium;

[5-(Hydroxy-diphenyl-methyl)-2H-pyrazol-3-ylmethyl]dimeth yl-(3- phenoxypropyl) ammonium;

[5-(Hydroxy-diphenyl-methyl) -1 H-[1 ,2,4]triazol-3-ylmethyl]-dimethyl-(3- phenoxypropyl)ammonium;

[2-(Hydroxy-diphenyl-methyl)-3-methyl-3H-imidazol-4-ylmet hyl]-dimethyl-(3- phenoxy-propyl)ammonium;

[3-(Cyc!ohexyl-hydroxy-phenyl-methyl)-isoxazol-5-ylmethyl ]-dimethyl-(3- phenoxy-propyl)-ammonium; [3-(Cyclohexyl-hydroxy-phenyl-methyl)-isoxazol-5-ylmethy!]-[ 2-(3,4-dichloro- benzyloxy)-ethyl]-dimethyl-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-isoxazol-5-ylmethyI ]-[3-(3,4-dichloro- phenoxy)-propyl]-dimethyl-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-isoxazol-5-ylmethyl ]-dimethyl-(2- phenethyloxy-ethyl)-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-isoxazol-5-ylmethyl ]-dimethyl-(4-phenyi-

butyl)-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-isoxazol-5-ylmethyl ]-[2-(3,4-dichloro- phenoxy)-ethyl]-dimethyl-ammonium;

(2-Benzyloxy-ethyl)-[3-(cyclohexyl-hydroxy-phenyl-methyl) -isoxazol-5-ylmethyl]- dimethyl-ammonium;

[2-(4-Chloro-benzyloxy)-ethyl]-[3-(cyclohexyl-hydroxy-phe nyl-methyl)-isoxazol- 5-ylmethyl]-dimethyl-ammonium;

[2-(3-Chloro-phenyl)-ethyl]-[3-(cyclohexyl-hydroxy-phenyl -methyl)-isoxazol-5- ylmethyl]-dimethyl-ammonium; [3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-5-ylmethyl]-[3-(3,4- dichloro-phenoxy)-propyl]-dimethyl-ammonium;

(2-Benzyloxy-ethyl)-[3-(cyclohexyl-hydroxy-phenyl-methyl) -[1 ,2,4]oxadiazol-5- ylmethyl]-dimethyl-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-5-ylmethyl]-[3-(2,3- dichloro-phenoxy)-propyl]-dimethyl-ammonium;

[2-(4-Chloro-benzyloxy)-ethyl]-[3-(cyclohexyl-hydroxy-phe nyl-methyl)- [1 ,2,4]oxadiazol-5-ylmethyl]-dimethyl-ammonium;

[3-(Cyclohexyl-hydroxy-ph ^' enyl-methyl)-[1 ,2,4]oxadiazol-5-ylmethyl]-dimethyl- (4-phenyl-butyl)-ammonium; [3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-5-ylmethyl]-dimethyl-

(2-phenethyloxy-ethyl)-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-5-ylmethyl]-[2-(3,4- dichloro-benzyloxy)-ethyl]-dimethyl-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-5-ylmethyl]-[2-(3,4- dichloro-phenoxy)-ethyl]-dimethyl-ammonium;

[2-(3-Chloro-phenyl)-ethyl]-[3-(cyclohexyl-hydroxy-phenyl -methyl)- [1 ,2,4]oxadiazol-5-ylmethyl]-dimethyl-ammonium;

[5-(Hydroxy-diphenyl-methyl)-[1 ,2,4]thiadiazol-3-ylmethyl]-dimethyl-(3-phenoxy- propyl)-ammonium; [5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]thiadiazol-3-ylmethyl]-dimethyl-

(3-phenoxy-propyl)-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]thiadiazol-5-ylmethyl]-dimethyl- (3-phenoxy-propyl)-ammonium;

^-(Cyclohexyl-hydroxy-phenyl-methyO-fi .S^lthiadiazol^-ylmethyll-dimethyl- (3-phenoxy-propyl)-ammonium;

[5-Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-3-ylmethyl]-[2-(3,4-

dichloro-phenoxy)-ethyl]-dimethyl-ammonium;

[2-(4-Chloro-benzyloxy)-ethyl]-[5-(cyclohexy!-hydroxy-phe nyl-methyl)- [1 ,2,4]oxadiazol-3-yImethyl]-dimethyl-ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-3-ylmethyl]-[2-(3,4- dichloro-benzyloxy)-ethyl]-dimethyl-ammonium;

^-(Cyclohexyl-hydroxy-phenyl-methyO-fi ^^loxadiazol-S-ylmethylJ-dinnethyl- (2-phenethyloxy-ethyl)ammonium;

[5-(Hydroxy-diphenyl-methyl)-isoxazol-3-ylmethyl]-dimethy l-(3-phenoxy-propyl)- ammonium; [5-(9-Hydroxy-9H-xanthen-9-yl)-[1 ,2,4]oxadiazol-3-ylmethyl]-dimethyl-(3- phenoxy-propyl)-ammonium;

Dimethyl-tδ-Cθ-methyl-θH-xanthen-θ-yO-ti ^^loxadiazol-S-ylmethylJ-CS- phenoxy-propyl)-ammonium;

[5-(-Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-[2-(3,4- dichloro-benzyloxy)-ethyl]-dimethyl-ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl- (2-p-tolyloxy-ethyl)-ammonium;

[3-(4-Chloro-phenylsuIfanyl)-propyl]-[5-(cyclohexyl-hydro xy-phenyl-methyl)- [1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl-ammonium; [5-(Cyclohexy!-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-[2-(2,3- dihydro-benzofuran-5-yl)-ethyl]-dimethyl-ammonium;

^-(Cyclohexyl-hydroxy-phenyl-methyO-ti .S^loxadiazol^-ylmethylJ-dimethyl- (4-methyl-pent-3-enyl)-ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl- (2-oxo-2-phenyl-ethyl)-ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-[3-(2- methoxycarbonyl-phenoxy)-propyl]-dimethyl-ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-(isoxazol-3- ylcarbamoylmethyl)-dimethyl-ammonium; [5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl-

[3-(4-sulfamoyl-phenoxy)-propyl]-ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl- [2-(4-trifluoromethyl-benzyloxy)-ethyl]-ammonium;

(3-Benzyloxy-propyl)-[5-(cyclohexyl-hydroxy-phenyl-methyl )-[1 ,3,4]oxadiazol-2- ylmethyl]-dimethyl-ammonium;

[3-(4-Carbamoyl-phenoxy)-propyl]-[5-(cyclohexyl-hydroxy-p henyl-methyl)-

[1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl-ammonium;

[3-(3-Chloro-4-cyano-phenoxy)-propyl]-[5-(cyclohexyI-hydr oxy-phenyl-methyl)- [1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl-ammonium;

[δ^Cyclohexyl-hydroxy-phenyl-methylJ-II .S.^oxadiazol^-ylmethyll-dimethyl- [2-(4-trifluoromethoxy-benzyloxy)-ethyl]-ammonium;

[2-(3-Chloro-4-methyl-benzyloxy)-ethyl]-[5-(cyclohexyl-hy droxy-phenyl-methyl)- [1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl-ammonium;

(R)-[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]- dimethyl-(4-methyl-benzyloxycarbonylmethyl)-ammonium; {2-[5-(Hydroxy-diphenyl-methyl)-isoxazol-3-yl]-ethyl}-dimeth yl-(3-phenoxy- propyl)-ammonium;

[2-(Cyclohexyl-hydroxy-phenyl-methyl)-3-methyl-3H-imidazo I-4-ylmethyl]- dimethyl-(3-phenoxy-propyl)-ammonium;

[2-(4-Chloro-benzyloxy)-ethyl]-[2-(cyclohexyl-hydroxy-phe nyl-methyl)-3-methyl- 3H-imidazol-4-ylmethyl]-dimethyl-ammonium;

[2-(Cyclohexyl-hydroxy-phenyl-methyl)-3-methyl-3H-imidazo l-4-ylmethyl]- dimethyl-(3-phenyl-propyl)-ammonium;

[2-(Cyclohexyl-hydroxy-phenyl-methyl)-3-methyl-3H-imidazo I-4-ylmethyl]- dimethyl-(3-phenyl-butyl)-ammonium; (4-Cyano-benzyl)-[2-(cyclohexyl-hydroxy-phenyl-methyl)-3-met hyl-3H-imidazol-

4-ylmethyl]-dimethyl-ammonium;

[2-(Cyclohexyl-hydroxy-phenyl-methyl)-3-methyl-3H-imidazo l-4-yimethyl]- dimethyl-(2-phenethyloxy-ethyl)-ammonium;

[2-(Cyclohexyl-hydroxy-phenyl-methyl)-3-methyl-3H-imidazo l-4-ylmethyl]- dimethyl-(4-p-tolyl-but-3-ynyl)-ammonium;

[2-(CycIohexyl-hydroxy-phenyl-methyl)-3-methyl-3H-imidazo l-4-ylmethyl] -dimethyl-(3-phenoxy-propyl)-ammonium; and

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-isoxazol-3-ylmethyl ]-dimethyl-(3- phenoxy-propyl)-ammonium; or pharmaceutically acceptable salts thereof.

In another aspect, the present invention provides compounds of formula (I) or (Ia) selected from:

[5-(Hydroxy-diphenyl-methyl)-1 ,3,4-oxadiazol-2-ylmethyl]-dimethyl-(3-phenoxy- propyl)-ammonium;

[3-(Hydroxy-diphenyl-methyl)-1 ,2,4-oxadiazol-5-ylmethyl]-dimethyl-(3-phenoxy-

propyl)-ammonium;

[5-(Hydroxy-diphenyl-methyl)-1 ,2,4-oxadiazol-3-ylmethyl]-dimethyl-(3-phenoxy- propyl)-ammonium;

[5-(Hydroxy-diphenyl-methyl)-thiazol-2-ylmethyl]-dimethyl -(3-phenoxy-propyI)- ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-5-yl methyl]-dimethyl- (3-phenoxyI-propyl)-ammonium;

[δ-CCyclohexyl-hydroxy-phenyl-methylHi ^^oxadiazol-S-yl methyl]-dimethyl- (3-phenoxyl-propyl)-ammonium; ^-(Cyclopentyl-hydroxy-phenyl-methyO-fi ,2,4]oxadiazol-3-yl methyl]-dimethyl-

(3-phenoxyl-propyl)-ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl- (3-phenoxy-propyl)-ammonium;

^-(Cyclopentyl-hydroxy-phenyl-methyO-fi .S^loxadiazol^-ylmethy^-dimethyl- (3-phenoxy-propyl)-ammonium;

[5-(Hydroxy-di-thiophen-2-yl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl-(3- phenoxy-propyl)-ammonium;

[3-Hydroxy- ^" diphenyl-methyl)-isoxazol-5-ylmethyl]-dimethyl-(3- phenoxypropyl)ammonium; [2-(Hydroxy-diphenyl-methyl)-3-methyl-3H-imidazol-4-ylmethyl ]-dimethyl-(3- phenoxy-propyl)ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-isoxazol-5-ylmethyl ]-dimethyl-(3- phenoxy-propyl)-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-isoxazol-5-ylmethyl ]-[2-(3,4-dichloro- benzyloxy)-ethyl]-dimethyl-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-isoxazol-5-ylmethyl ]-[3-(3,4-dichloro- phenoxy)-propyl]-dimethyl-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-isoxazol-5-ylmethyl ]-dimethyl-(2- phenethyloxy-ethyl)-ammonium; [3-(Cyclohexyl-hydroxy-phenyl-methyl)-isoxazol-5-ylmethyl]-d imethyl-(4-phenyl- butyl)-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-isoxazol-5-ylmethyl ]-[2-(3,4-dichloro- phenoxy)-ethyl]-dimethyl-ammonium;

(2-Benzyloxy-ethyl)-[3-(cyclohexyl-hydroxy-phenyl-methyl) -isoxazol-5-ylmethyl]- dimethyl-ammonium;

[2-(4-Chloro-benzyloxy)-ethyl]-[3-(cyclohexyl-hydroxy-phe nyl-methyl)-isoxazol-

5-ylmethyl]-dimethyl-ammonium;

[2-(3-Chloro-phenyl)-ethyl]-[3-(cyclohexyl-hydroxy-phenyl -methyl)-isoxazol-5- ylmethyl]-dimethyl-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-5-ylmethyl]-[3-(3,4- dichloro-phenoxy)-propyl]-dimethyl-ammonium;

(2-Benzyloxy-ethyl)-[3-(cyclohexyl-hydroxy-phenyl-methyl) -[1 ,2,4]oxadiazol-5- ylmethyl]-dimethyl-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-5-ylmethyl]-[3-(2,3- dichloro-phenoxy)-propyl]-dimethyl-ammonium; [2-(4-Chloro-benzyloxy)-ethyl]-[3-(cyclohexyl-hydroxy-phenyl -methyl)-

[1 ,2,4]oxadiazol-5-ylmethyl]-dimethyl-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-5-ylmethyl]-dimethyl- (4-phenyl-butyl)-ammonium;

^-(Cyclohexyl-hydroxy-phenyl-methyO-CI ^^Joxadiazol-B-ylmethylj-dimethyl- (2-phenethyIoxy-ethyl)-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyI)-[1 ,2,4]oxadiazol-5-ylmethyl]-[2-(3,4- dichloro-benzyloxy)-ethyl]-dimethyl-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-5-ylmethyl]-[2-(3,4- dichloro-phenoxy)-ethyl]-dimethyl-ammonium; [2-(3-Chloro-phenyl)-ethyl]-[3-(cyclohexyl-hydroxy-phenyl-me thyl)-

[1 ,2,4]oxadiazol-5-ylmethyl]-dimethyl-ammonium;

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]thiadiazol-5-ylmethyl]-dimethyl- (3-phenoxy-propyl)-ammonium;

^-(Cyclohexyl-hydroxy-phenyl-methylJ-fi .S^Jthiadiazol^-ylmethyn-dimethyl- (3-phenoxy-propyl)-ammonium;

[5-CyclohexyI-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-3-ylmethyl]-[2-(3,4- dichloro-phenoxy)-ethyl]-dimethyl-ammonium;

[2-(4-Chloro-benzyloxy)-ethyl]-[5-(cyclohexyl-hydroxy-phe nyl-methyl)- [1 ,2,4]oxadiazol-3-ylmethyl]-dimethyl-ammonium; [5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-3-ylmethyl]-[2-(3,4- dichloro-benzyloxy)-ethyl]-dimethyl-ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-3-ylmethyl]-dimethyl- (2-phenethyloxy-ethyl)ammonium;

[5-(Hydroxy-diphenyl-methyl)-isoxazol-3-ylmethyl]-dimethy l-(3-phenoxy-propyl)- ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-[2-(3,4-

dichloro-benzyloxy)-ethyl]-dimethyl-ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl- (2-p-tolyloxy-ethyl)-ammonium b;

[3-(4-Chloro-phenylsulfanyl)-propyl]-[5-(cyclohexyl-hydro xy-phenyl-methyl)- [1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl-ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-[2-(2,3- dihydro-benzofuran-5-yl)-ethyl]-dimethyl-ammonium;

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl- [2-(4-trifluoromethyl-benzyloxy)-ethyl]-ammonium; (3-Benzyloxy-propyl)-[5-(cyclohexyl-hydroxy-phenyl-methyl)-[ 1 ,3,4]oxadiazol-2- ylmethyl]-dimethyl-ammonium;

[5-(CyclohexyI-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethy!]-dimethyl- [2-(4-trifluoromethoxy-benzyloxy)-ethyl]-ammonium;

[2-(3-Ch!oro-4-methyl-benzyloxy)-ethyl]-[5-(cyclohexyl-hy droxy-phenyl-methyl)- [1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl-ammonium; and

(R)-[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]- dimethyl-(4-methyl-benzyloxycarbonylmethyl)-ammonium; or pharmaceutically acceptable salts thereof.

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 (also known as chronic obstructive pulmonary disease or COPD), chronic bronchitis of all types

(including dyspnoea associated therewith), asthma (allergic and non-allergic; 'wheezy- infant syndrome'), adult/acute respiratory distress syndrome (ARDS), chronic respiratory obstruction, bronchial hyperactivity, pulmonary fibrosis, pulmonary emphysema, and allergic rhinitis, exacerbation of airway hyperreactivity consequent to other drug therapy, particularly other inhaled drug therapy, 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.

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

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, diluent or excipient.

Another aspect of the invention provides a compound of formula (I), (Ia) or (Ib) for use in therapy.

Another aspect of the invention is the use of a compound of formula (I) or (Ia) 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 provides a method of treatment of a disease or

condition in which M3 muscarinic receptor activity is implicated comprising administration to a subject in need thereof of an effective amount of a compound of formula (I) or (Ia), or a pharmaceutically acceptable salt thereof.

Another aspect of the invention is the use of a compound of formula (I) or (Ib), in the manufacture of a medicament for the treatment of prevention of a disease or condition in which M3 muscarinic receptor activity and β-adrenergic activity is ^■ implicated.

Another aspect of the invention provides a method of treatment of a disease or condition in which M3 muscarinic receptor activity and β-adrenergic activity are implicated comprising administration to a subject in need thereof of an effective amount of a compound of formula (I) or (Ib), or a pharmaceutically acceptable salt thereof.

Description of Definitions

Unless otherwise qualified in the context in which they are used, the following terms have the following meanings when used herein:

"Acyl" means a -CO-alkyl group in which the alkyl group is as described herein. Exemplary acyl groups include -COCH ₃ and -COCH(CH ₃ ) ₂ .

"Acylamino" means a -NR-acyl group in which R and acyl are as described herein. Exemplary acylamino groups include -NHCOCH ₃ and -N(CH ₃ )COCH ₃ .

"Alkoxy" and "alkyloxy" means an -O-alkyl group in which alkyl is as described below. Exemplary alkoxy groups include methoxy (-OCH ₃ ) and ethoxy (-OC ₂ H ₅ ). ' "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, more conveniently 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, more conveniently 2 to 6, carbon atoms and one or more carbon-carbon double bonds in the chain; more conveniently 1 , 2 or 3 carbon-carbon double bonds in the chain; more conviently still, one carbon-carbon , double bond in the chain. Exemplary alkenyl groups include ethenyl, 1 -propenyl, and 2-propenyl.

"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 -CH ₂ -, -(CH ₂ ) ₂ - and -C(CH ₃ )HCH ₂ -. "Alkenylene" means an -alkenyl- group in which alkenyl is as defined previously. Exemplary alkenylene groups include -CH=CH-, -CH=CHCH ₂ -, and - CH ₂ CH=CH-.

"Alkynylene" means an -alkynyl- group in which -alkynyl- refers to a straight or branched chain hydrocarbon group having from 2 to 12, more conveniently 2 to 6, carbon atoms and one, two or three carbon-carbon triple bonds in the chain; or, more conveniently, one carbon-carbon triple bond in the chain. Exemplary alkynylene groups include ethynyl and propargyl.

"Alkylsulfinyl" means a -SO-alkyl group in which alkyl is as defined above. Exemplary alkylsulfinyl groups include methylsulfinyl and ethylsulfinyl. "Alkylsulfonyl" or "sulfonyl" each means a -SO ₂ -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 -CONH ₂ and -CONHCH ₃ .

"Aminoalkyl" means an alkyl-NH ₂ group in which alkyl is as previously described. Exemplary aminoalkyl groups include -CH ₂ NH ₂ .

"Aminosulfonyl" means a -SO ₂ -NRR group in which R is as herein described. Exemplary aminosulfonyl groups include -SO ₂ NH ₂ and -SO ₂ NHCH ₃ .

"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, more conveniently from 6 to 10 carbon atoms, such as phenyl or naphthyl. Phenyl is a specifically preferred aryl group. The aryl group, specifically a phenyl 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.

"Arylalkyloxy" means an aryl-alkyloxy- group in which the aryl and alkyloxy moieties are as previously described. Preferred arylalkyloxy groups contain a C _{1 4} alkyl moiety. Exemplary arylalkyl groups include benzyloxy.

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

"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, benxodiόxolyl, 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.

"Cyclic amine" 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, more conveniently from 3 to 8 carbon atoms, and more conveniently still 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, more conveniently 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 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. "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.

"Heteroaryloxy" means a heteroaryloxy- group in which the heteroaryl is as previously described. Exemplary heteroaryloxy groups include pyridyloxy. "Heteroaryl-fused-cycloalkyl" means a monocyclic heteroaryl group, such as pyridyl or furanyl, fused to a cycloalkyl group, in which heteroaryl and cycloalkyl are as previously described. Exemplary heteroaryl-fused-cycloalkyl groups include tetrahydroquinolinyl and tetrahydrobenzofuranyl. The heteroaryl and cycloalkyl rings may each be substituted by one or more substituent groups. The heteroaryl-fused- cycloalkyl group may be attached to the remainder of the compound by any available carbon or nitrogen atom.

"Heteroaryl-fused-heterocycloalkyl" means a monocyclic heteroaryl group, such as pyridyl or furanyl, fused to a heterocycloalkyl group, in which heteroaryl and heterocycloalkyl are as previously described. Exemplary heteroaryl-fused- heterocycloalkyl groups include dihydrodioxinopyridinyl, dihydropyrrolopyridinyl, dihydrofuranopyridinyl and dioxolopyridinyl. The heteroaryl and heterocycloalkyl rings may each be substituted by one or more substituents groups. The heteroaryl-fused- heterocycloalkyl group may be attached to the remainder of the compound by any available carbon or nitrogen atom. "Heterocycloalkyl" 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. CON R or CONRCO (examples of such groups include succinimidyl and 2-oxopyrrolidinyl). The heterocycloalkyl group may be substituted by one or more substituent 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). "Sulfonylamino" means a -NR-sulfonyl group in which R and sulfonyl are as described herein. Exemplary sulfonylamino groups include -NHSO ₂ CH ₃ .

A substituent designatation R in any of the above definitions means hydrogen, alkyl, aryl, or heteroaryl as described herein, and when two R groups are present on a group (for example on -SO _2- NRR) then the R groups can be the same or different. "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, maleates, fumarates, succinates and the like; (iii) for the quaternary ammonium group of a compound of formula (I) or (Ia) an acceptable couriter-ion (D ^' ) 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 pharmaceutically acceptable counter-ion D ^" such that compound of formula (Ia) has no net charge.

The present invention covers all permissible ratios of cationic ammonium species to anion D ^" , for example hemi-napadisylate and napadisylate.

In another aspect of the invention D ^" is bromide or napadisylate (for example naphthalene-1 ,5-disulfonate).

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. Suitable groups for forming pro-drugs are described in The Practice of Medicinal Chemistry, 2 ^nd Ed. pp561-585 (2003) and in F. J. Leinweber, Drug Metab. Res., , 18, 379. (1987)

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.

The cyclic groups referred to above, namely, aryl, heteroaryl, cycloalkyl, aryl- fused-cycloalkyl, heteroaryl-fused-cycloalkyl, heterocycloalkyl, aryl-fused- heterocycloalkyl, heteroaryl-fused-heterocycloalkyl and cyclic amine are unsubstituted or substituted by one or more of the same or different substituent groups. Examples of specific optional substituents include -Cl, -F, -CH ₃ , -CF ₃ , -OCH ₃ , -OH, -CN, -COOCH ₃ , -CONH ₂ , -SO ₂ NH ₂ , -SO ₂ N(CH ₃ ) ₂ . More generally the substituents can be divided into two classes: (a) a first class of substituent includes acyl (e.g. -COCH ₀ ), alkoxy (e.g., -

OCH ₃ ), alkoxycarbonyl (e.g. -COOCH ₃ ), alkylamino (e.g. -NHCH ₃ ), alkylsulfinyl (e.g. -SOCHJ o , alkylsulfonyl (e.g. -SO ₀ 2CH oJ, alkylthio (e.g. -SCHJ o , -NH ₂ , aminoacyl (e.g. -

CON(CHg) ₂ ), aminoalkyl (e.g. -CH ₀ NHJ, cyano, dialkylamino (e.g. -N(CHJJ, halo, haloalkoxy (e.g. -OCF, or -OCHFJ, haloalkyl (e.g. -CFJ, alkyl (e.g. -CH or -CH CHJ, -OH, -CHO, -COOH, -NO ₂ , aminoacyl (e.g. -CONH ₂ , -CONHCH ₃ ), aminosulfonyl (e.g. -SO ₂ NH ₂ , -SO ₂ NHCH ₃ ), acylamino (e.g. -NHCOCH ₃ ) and sulfonylamino (e.g. - NHSO ₂ CH ₃ ); and

(b) a second class of substituent includes arylalkyl (e.g. -CH ₂ Ph or

-CH ₂ -CH ₂ -Ph), aryl, heteroaryl, heterocycloalkyl, heteroarylalkyl, cyclic amine (e.g. morpholine), aryloxy, heteroaryloxy, arylalkyloxy (e.g. benzyloxy) and

heteroarylalkyloxy, the cyclic part of any of which being optionally substituted by any of the first class of substituent referred to above (for example alkoxy, haloalkoxy, halogen, alkyl and haloalkyl). ^'

In one aspect, alkyl, alkoxy, alkylene, alkenyl or alkenylene groups may be optionally substituted. In another aspect, alkylene, alkenyl or alkenylene groups may be optionally substituted. Suitable optional substituent groups for both these aspects include alkoxy (e.g., -OCH ₃ ), alkylamino (e.g. -NHCH ₃ ), alkylsulfinyl (e.g. -SOCH ₃ ), alkylsulfonyl (e.g. -SO ₂ CH ₃ ), alkylthio (e.g. -SCH ₃ ), -NH ₂ , aminoalkyl (e.g. -CH ₂ NH ₂ ), arylalkyl (e.g. -CH ₂ Ph or -CH ₃ -CH ₂ -Ph), cyano, dialkylamino (e.g. -N(CH ₃ ) ₂ ), halo, haloalkoxy (e.g. -OCF ₃ or -OCHF ₂ ), haloalkyl (e.g. -CF ₃ ), alkyl (e.g. -CH ₃ or -CH ₂ CH ₃ ),

-OH, -CHO, and -NO ₂ .

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-torms, 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 groups R ¹ , R ² and R ³

In one aspect, the present invention provides compounds of formula (I) and (Ia) wherein the groups R ¹ , R ² and R ³ are combined so that:

R ¹ is d-Cβ-alkyl; and R ² is a group (Z) _p -R ⁷ , -Z-Y-R ⁷ , -Z-NR ⁹ R ¹⁰ , -Z-CO-NR ⁹ R ¹⁰ , or - Z-C(O)-R ⁷ ; and R ³ is C _r C ₆ -alkyl; p is O or 1 ; provided that R ¹ , R ² and R ³ do not all represent CH ₃ .

In another aspect, the present invention provides compounds of formula (Ib) wherein the groups R ¹ and R ³ are combined so that:

R ¹ is Ci-C ₆ -alkyl; and R ³ is a lone pair or C _r C ₆ -alkyl.

In a further aspect, where a group (Z) _p -R ⁷ , or -Y-R ⁷ , -Z-Y-R ⁷ , or a group -Z-NR ⁹ R ¹⁰ ,

or a group -Z-CO-NR ⁹ R ¹⁰ , or a group -Z-C(O)-R ⁷ , is present in R ² (provided that R ¹ , R ² and R ³ do not all represent CH ₃ and that when R ³ is a lone pair then R ¹ and R ² do not both represent CH ₃ ):

Z may be, for example -(CH ₂ )M ₆ - optionally substituted on up to three carbons in the chain by methyl;

Y is -O- or a group -S(O) _n (where n is 0,1 or 2), C(O)O, OC(O), N(R ¹² )S(O) ₂ Or S(O) ₂ N(R ¹² );

R ⁷ may be

CrCβ-alkyl, such as methyl, ethyl, n- or isopropyl, n-, sec- or tertbutyl;

Optionally substituted aryl such as phenyl or naphthyl, or aryl-fused- heterocycloalkyl such as 3,4-methylenedioxyphenyl, 3,4- ethylenedioxyphenyl, or dihydrobenzofuranyl;

Optionally substituted heteroaryl such as pyridyl, pyrrolyl, pyrimidinyl, oxazolyl, isoxazolyl, benzisoxazolyl, benzoxazolyl, thiazolyl, benzothiazolyl, quinolyl, thienyl, benzothienyl, furyl, benzofuryl, imidazolyl, benzimidazolyl, isothiazolyl, benzisothiazolyl, pyrazolyl, isothiazolyl, triazolyl, benzotriazolyl, thiadiazolyl, oxadiazolyl, pyridazinyl, pyridazinyl, triazinyl, indolyl and indazolyl;

Optionally substituted ary^CrCe-alkyl)- such as those wherein the aryl part is any of the foregoing specifically mentioned aryl groups and the

-(C _r C ₆ -alkyl)- part is -CH ₂ - or -CH ₂ CH ₂ -;

Optionally substituted heteroaryl(CrC ₈ -alkyl) ^' - such as those wherein the heteroaryl part is any of the foregoing specifically mentioned heteroaryl groups and the -(Ci-C ₆ -alkyl)- part is -CH ₂ - or -CH ₂ CH ₂ -;

Optionally substituted cycloalkyl such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, or move conveniently, such as cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl;

More conveniently, R ⁷ may be

Optionally substituted aryl such as phenyl or naphthyl, or aryl-fused- heterocycloalkyl such as 3,4-methylenedioxyphenyl, 3,4- ethylenedioxyphenyl, or dihydrobenzofuranyl; or

Optionally substituted ary^CrC _θ -alkyl)- such as those wherein the aryl part is any of the foregoing specifically mentioned aryl groups and the -(C _r C ₆ -alkyI)- part is -CH ₂ - or -CH ₂ CH ₂ -.

R ⁹ and R ¹⁰ may be independently selected from hydrogen; C _r C ₆ -alkyl such as methyl, ethyl or n- or isopropyl; or any of those optionally substituted aryl, aryl- fused-heterocycloalkyl, heteroaryl or aryl(CrC ₈ -alkyl)- groups specifically mentioned in the discussion of R ⁷ above; or

R ⁹ and R ¹⁰ together with the nitrogen atom to which they are attached may form a heterocyclic ring of 4-8 ring atoms, more conveniently 4-6 ring atoms, optionally containing a further nitrogen or oxygen atom, such as azetidinyl, piperidinyl, piperazinyl, N-substituted piperazinyl such as methylpiperazinyl, pyrrolidinyl, morpholinyl, and thiomorpholinyl.

In a yet further aspect, the present invention provides compounds of formula (I) and (Ia) wherein R ² may represent an optionally substituted ary^CrCe-alkyl)- moiety such as, for example, a moiety wherein the aryl part is optionally substituted phenyl (for example, 3-chlorophenyl) and the -(C^Ce-alkyl)- part is, for example, -CH ₂ CH ₂ -.

One aspect of the present invention provides compounds of formula (I) and (Ia) wherein, R ¹ is methyl or ethyl, R ² is -Z-Y-R ⁷ as defined and discussed above, Y is -O- and -Z- is a straight or branched alkylene radical linking the nitrogen and -YR ⁷ by a chain of up to 16, for example up to 10, and more conveniently from 2 to 4, carbon atoms, and R ³ is methyl. In these cases, R ⁷ is conveniently a cyclic lipophilic group such as phenyl, benzyl, dihydrobenzofuranyl or phenylethyl (wherein the phenyl rings are optionally substituted as described above).

Another aspect of the present invention provides compounds of formula (I) and (Ia) wherein, R ¹ is methyl or ethyl, R ² is -(Z) _p -R ⁷ as defined and discussed above, p is 1 , and -Z- is a straight or branched alkylene radical linking the nitrogen and -R ⁷ by a

chain of up to 16, for example up to 10, and more conveniently from 2 to 5, carbon atoms, and R ³ is methyl. In these cases, R ⁷ is conveniently a cyclic lipophilic group such as phenyl or dihydrobenzofuranyl (wherein the phenyl rings are optionally substituted as described above).

The groups R ⁴ . R ⁵ and R ⁶

R ⁴ and R ⁵ may be independently selected from any of those aryl, aryl-fused- heterocycloalkyl, aryl-fused-cycloalkyl, heteroaryl, C _r C ₆ -alkyl, or cycloalkyl groups specifically mentioned in the discussion of R ⁵ above. R ⁶ may be -OH, a hydrogen atom, C _r C ₆ -alkyl such as methyl or ethyl, CrC ₆ -alkoxy such as methoxy or ethoxy, hydroxy-CrC ₆ -alkyl such as hydroxymethyl, nitrile, or a group CONR ¹³ ₂ wherein each R ¹³ is independently CrC ₆ -alkyl such as methyl or ethyl, or a hydrogen atom. Most convenient is the case where R ⁶ is -OH. Convenient combinations of R ⁴ and R ⁵ , especially when R ⁶ is -OH, include those wherein (i) each of R ⁴ and R ⁵ 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) each of R ⁴ and R ⁵ is optionally substituted phenyl; (iii) one of R ⁴ and R ⁵ is optionally substituted phenyl and the other is cycloalkyl such as cyclopropyl, cyclobutyl, or especially cyclopentyl or cyclohexyl; or more conveniently, cycloalkyl such as cyclobutyl, cycloheptyl, cyclooctyl or especially cyclopentyl or cyclohexyl ; (iv) one of R ⁴ and R ⁵ 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 or cyclohexyl; and (v) R ⁴ and R ⁵ are joined together to form a tricyclic ring so that the

group R ⁴ R ⁵ R ⁶ C- represents the group , where R ^6a is -OH, C _r C ₆ -alkyl or a hydrogen atom, and Q is an oxygen atom, -CH ₂ -, -CH ₂ CH ₂ - or a bond. Most convenient are the combinations of R ⁴ and R ⁵ represented by options (ii) and (iii). When R ⁶ is -OH and R ⁴ and R ⁵ are represented by option (iii), conveniently the carbon to which R ⁴ , R ⁵ and R ⁶ are attached has the (R)- absolute configuration.

W, V, A

The permitted and specific combinations of W, V and A in compounds of formula (I),

(Ia) and (Ib) have been defined and specified above.

The group R ¹¹ may be selected from a hydrogen atom or a C _r C ₃ -alkyl, especially a methyl group.

The Radical X

Although X may be an alkylene, alkenylene or alkynylene radical, it is currently preferred that it be alkylene, for example ethylene or methylene.

The Radical Lf The permitted combinations of L ₁ L ¹ , L ² , L ³ and L ⁴ are defined above. Currently preferred is that L ¹ , L ² , L ³ and L ⁴ are hydrogen or C _1-3 alkyl. Up to three of the carbon atoms of the hydrocarbyl group L may be substituted with groups as defined above. It is currently preferred that the group L is an alkylene chain, more conveniently C ₅ -C ₇ - alkylene.

The Radical Z ¹

The radical Z ¹ is a β agonist group as defined above. In groups of formula (A), it is convenient for the group Ar to be (i) a 4-hydroxy-3-hydroxymethyl-phenyl group, (ii) a 3-formylamino-4-hydroxy-phenyl group, or especially, (iii) an 8-hydroxy-2-oxo-1 ,2- dihydroquinolinyl group or (iv) a 4-hydroxy-2-oxo-2,3-dihydro-benzothiazolyl group.

A subclass of compounds with which the invention is concerned consists of those of formula (ID)

wherein B is a 5-membered heterocyclic ring selected from the group:

R -.1"1 « r,11

R ¹ X

N-O N-N N-N N-N

N N N

wherein the bond marked ^* is attached to the group R ⁴ R ⁵ (OH)C-, and the bond marked ^** is attached to the group -(CH ₂ ) _m -; m is 1 or 2; ring Q is an optionally substituted phenyl ring, or phenyl-fused- heterocycloalkyl ring system wherein the heterocycloalkyl ring is a monocyclic heterocyclic ring of 5 or 6 ring atoms; R ⁴ is phenyl or thienyl; R ⁵ is phenyl; thienyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl; s is 2, 3, 4, 5, 6 or 7 and t is 0, 1 , 2, 3, 4, 5, 6 or 7; Y is -O-, a group -S(O) _n , C(O)O, OC(O), N(R ¹² )S(O) ₂ or S(O) ₂ N(R ¹² ); n is 0,1 or 2, and D ^" is a pharmaceutically acceptable anion.

In one aspect the present invention provides a compound of formula (ID) wherein Q is optionally substituted phenyl (wherein optional substituents are selected from alkoxy, halo (such as fluoro or chloro), Ci-C ₃ -alkyl, cyano, -CF ₃ , amino C _r C ₃ -acyl or amino Ci-C ₃ -alkyl; or more conveniently optional substituents are selected from alkoxy, halo (such as fluoro or chloro), C _r C ₃ -alkyl, cyano, or -CF ₃ or a phenyl-fused- heterocycloalkyl ring system (wherein the heterocycloalkyl ring is a monocyclic heterocyclic ring of 5 or 6 ring atoms, such as dihydrobenzofuranyl).

In subclass (ID), s+t is, for example, 2, 3, 4, 5, 6, or 7 and may arise from suitable combinations of t and s such as where t is 0, 1 , 2, 3, 4, 5 or 6 and s is 2, 3, 4, 5, 6 or 7.

In compounds (ID) a combination of t, Y and s is where t is O, s is 3, and Y is -O. A further combination of t, Y and s is where t is 1 , s is 2 and Y is -O-. A yet further combination of t, Y and s is where t is 2, s is 2 and Y is -0-.

In one aspect m is 1 for compounds of (ID). In another aspect, m is 2.

Another subclass of compounds with which the invention is concerned consists of those of formula (IE)

wherein B is as defined above, m is 1 or 2; ring Q is an optionally substituted phenyl ring, or monocyclic heterocyclic ring of 5 or 6 ring atoms, or phenyl-fused- heterocycloalkyl ring system wherein the heterocycloalkyl ring is a monocyclic heterocyclic ring of 5 or 6 ring atoms; R ⁴ is phenyl or thienyl; R ⁵ is phenyl; thienyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl; u is 1 , 2, 3, 4, 5, and D ^" is a pharmaceutically acceptable anion.

In one aspect the present invention provides a compound of formula (IE) wherein Q is optionally substituted phenyl (wherein optional substituents are selected from alkoxy, halo (such as fluoro or chloro), Ci-C ₃ -alkyl, -CN, -CF ₃ or a phenyl-fused- heterocycloalkyl ring system (wherein the heterocycloalkyl ring is a monocyclic heterocyclic ring of 5 or 6 ring atoms, such as dihydrobenzofuranyl).

In one aspect m is 1 for compounds of (IE).

As previously mentioned, it will be appreciated that certain combinations of R ⁴ , R ⁵ and R ⁶ can give rise to optical enantiomers. In such cases, both enantiomers of the invention generally exhibit affinity at the M ₃ receptor, although the skilled person will appreciate that one enantiomer may display enhanced potency at the M ₃ receptor and/or selectivity against the M ₂ receptor.

Examples of compounds of the invention include those of the Examples herein.

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; Other muscarinic receptor antagonists; 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 β2-adrenoreceptor agonist, for example albuterol (salbutamol), salmeterol, metaproterenol, terbutaline, fenoterol, procaterol, carmoterol, indacaterol, formoterol, arformoterol, picumeterol, GSK-159797, GSK-597901 , GSK-159802, GSK-64244, GSK-678007, TA-2005 and also compounds of EP1440966, JP05025045, WO93/18007, WO99/64035, US2002/0055651 , US2005/0133417, US2005/5159448, WO00/0751 14, WO01/42193, WO01 /83462, WO02/66422, WO02/70490, WO02/76933, WO03/24439, WO03/42160, WO03/42164, WO03/72539, WO03/91204, WO03/99764, WO04/16578, WO04/016601 , WO04/22547, WO04/32921 , WO04/33412, WO04/37768, WO04/37773, WO04/37807, WO0439762, WO04/39766, WO04/45618, WO04/46083, WO04/71388, WO04/80964, EP1460064, WO04/087142, WO04/89892, EP01477167,

US2004/0242622, US2004/0229904, WO04/108675, WO04/108676, WO05/033121 ,

WO05/040103, WO05/044787, WO04/071388, WO05/058299, WO05/058867,

WO05/065650, WO05/066140, WO05/070908, WO05/092840, WO05/092841 ,

WO05/092860, WO05/092887, WO05/092861 , WO05/090288, WO05/092087,

WO05/080324, WO05/080313, US20050182091 , US20050171147, WO05/092870, WO05/077361 , DE10258695, WO05/111002, WO05/111005, WO05/110990,

US2005/0272769 WO05/110359, WO05/121065, US2006/0019991 , WO06/016245,

WO06/014704, WO06/031556, WO06/032627, U S2006/0106075, US2006/0106213,

WO06/051373, WO06/056471 ;

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 EP1241 176;

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 PGD ₂

(DP1 or CRTH2), or a thromboxane A ₂ antagonist eg ramatrobant; 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.

A further aspect of the present invention provides a combination comprising a compound of formula (Ia), as defined herein above, and a β2-adrenoreceptor agonist.

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 1 μg 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). Propellants suitable for use in a PMDI are known to the skilled person, and include CFC-12, HFA-134a, HFA-227, HCFC-22 (CCI ₂ F ₂ ) and HFA-152 (C ₂ H ₄ F ₂ ) 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 compounds of the invention of the present invention can be prepared according to the procedures of the following schemes and examples, using appropriate materials, and are further exemplified by the following specific examples. Moreover, by utilising the procedures described with the disclosure contained herein, one of ordinary skill in the art can readily prepare additional compounds of the present

invention claimed herein. The compounds illustrated in the examples are not, however, to be construed as forming the only genus that is considered as the invention. The examples further illustrate details for the preparation of the compounds of the present invention. Those skilled in the art will readily understand that known variations of the conditions and processes of the following preparative procedures can be used to prepare these compounds.

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

The invention further provides a process for the preparation of a compound of formula (I) or (Ia) or a pharmaceutically acceptable salt thereof, as defined above, according to the routes illustrated in Schemes 1 -17.

= NH, V = N , A = N W = NH, V = N ₁ A = N W = NH ₁ V = N, A = = N

W = N, V = N, A = O W = N ₁ V = N, A = O W = N ₁ V = N ₁ A = O

R ² D

W = NH, V = N, A = N

W = N, V = : N, A = O

Scheme 1

Scheme 2

(R ⁴ not equal R ⁵ )

R ³ D

1. DIBAL-H

2. R ⁴ MgBr V = O ₁ A=N ₁ W = C;

3. PCC V = S, A = N, W = C; W-V

V = N ₁ A = O ₁ W = C; Ph D

4. R ⁵ MgBr

V = N ₁ A=S ₁ W = C; HO

V = O ₁ A=C ₁ W = N; Ph

V = N ₁ A=N ₁ W = S R _R 3

Scheme 3

Scheme 4

Scheme 5

Scheme 6

HNR ¹ R ²

Scheme 7

Scheme 8

Scheme 9

W = CV = N ₁ A = S ^* W = C ₁ V = N ₁ A = S W = C ₁ V = N ₁ A=S W = N ₁ V = C ₁ A = N-Me W = N ₁ V = C ₁ A=N-Me

(1) SOCI ₂ R ³ D

(2) MeNH ₁ CHCI ₃

W= N ₁ V= C, A = N-Me V = N ₁ W = C, A = S W = N ₁ V = C ₁ A = N-Me

Scheme 10

Scheme 11

OH

(1) CBr ₄ , PPh ₃

Scheme 12

Et ₃ SiH, TFA R* = C« alkyl

Scheme 13

4. HNR ¹ R ² NaBH(OAc) ₃

Scheme 14

Scheme 15

Scheme 16

Scheme 17

It will be recognised by those skilled in the art that in Schemes 1 -17 above where none of R ¹ , R ² and R ³ are a lone pair or hydrogen atom (and the nitrogen to which they are attached carries a positive charge), the introduction of groups R ¹ , R ² and R ³ may be achieved in a different order, i.e. generally any of R ¹ , R ² and R ³ an be the quaternising group. Additionally in some cases the quaternary ammonium compound may be accessed directly by treatment of a suitable intermediate with a trtiary amine NR ¹ R ² R ³ .

In a further aspect, the present invention provides a process for the preparation of a compound of formula (Ia), as previously defined, comprising the reaction of a compound of formula (XXIX):

wherein R ¹ , R ² , R ⁴ , R ⁵ , R ⁶ , W, V, A and X are as previously defined, with a compound of formula (XXX):

R ³ D (XXX) wherein R ³ and D are as previously defined, at a suitable temperature, optionally in the presence of a suitable solvent (such as acetonitrile, methylene chloride, chloroform, tetrahydrofuran, isopropanol or mixtures thereof).

When solvent(s) is used, the reaction may be carried out at a temperature between

0 ⁰ C and the reflux temperature of the solvent(s). ,

When no solvent is used, the reaction may be carried out at a temperature between ambient temperature and 150 ⁰ C or, more conveniently, between 50 and 100 ⁰ C.

The invention further provides a process for the preparation of a compound of formula ( (II)) oorr ((IIbb)) oorr aa pphhaarrmmaacceeuuttiiccaallllyy aacccceeppttaabbllee ssaalltt tthheerreeoof, as defined above, according to one of the general routes illustrated in Scheme 18;

(VII-I)

Scheme 18

Compounds of formula (Ib-i) wherein R ⁴ , R ⁵ , R ⁶ , W, V, A, X, R ¹ , R ³ , L ¹ , L ² , L ³ , L ⁴ , L and Ar are as defined above for compound of formula (Ib) and T is a hydrogen atom

or hydroxy group (optionally suitably protected), may be prepared; from compounds of formula (lll-i) by reaction with a suitable electrophile, i.e by reaction with a compound of formula (ll-i) wherein LG is a suitable leaving group under alkylation conditions; by reaction with a compound of formula (IV-i) under reductive amination conditions or by coupling with a carboxylic acid or acid derivative of formula (XX-i) followed by reduction of the amide bond; from compounds of formula (Vll-i) by reaction with a suitable electrophile, i.e by reaction with a compound of formula (Vl ll-i) wherein LG is a suitable leaving group under alkylation conditions; when T = OH, by reaction with a compound of formula (IX-i), or with a compound of formula (X-i) followed by reduction of the carbonyl group; or from compounds of formula (Xlll-i) by reaction with a suitable electrophile, i.e. by reaction with a compound of (XIV-i) wherein LG is a suitable leaving group under alkylation conditions; by reaction with a compound of formula (XII-i) under reductive amination conditions, or by coupling with a carboxylic acid or acid derivative of formula (Xll-ii) followed by reduction of the amide bond; or from compounds of formula (XlV-ii) by reaction with a suitable electrophile, i.e. by reaction with a compound of formula (XXVII-i) under alkylation conditions, or with a compound of formula (XXViI-H) under reductive amination conditions, wherein b is a suitable value to provide X.

Where described above, 'alkylation conditions' refer to reaction of an amine with an electrophile containing a suitable leaving group (such as bromide, chloride, iodide or a sulfonate) in a suitable comaptible solvent, optionally in the presence of a base such as a metal carbonate or hydrogen carbonate, or an organic base such as triethylamine, diisopropyethylamine or pyridine, at a temperature from O ⁰ C to the reflux temperature of the solvent.

Where described above, 'reductive amination conditions' refer to reaction of an amine with a suitable aldehyde or ketone containing compound in the presence of a reducing agent such as a borohydride, for example sodium triacetoxyborohydride or sodium cyanoborohydride, or hydrogen in the presence of a suitable catalyst, for example palladium on carbon or platinum oxide, in the presence of a compatible solvent, at a temperature from 0 ⁰ C to the reflux temperature of the solvent, preferably from O ⁰ C to ambient temperature.

Where described above, 'reduction of the amide bond' may be achieved using borane or borane complexes, borohydrides such as sodium or lithium borohydride, or lithium aluminium hydride, in a compatible solvent at a temperature from -20 ⁰ C to the

reflux temperature of the solvent, preferably from O ⁰ C to the reflux temperature of the solvent.

Where described above, 'coupling with a carboxylic acid or acid derivative' refers to the formation of an amide bond either employing an acid and suitable coupling reagent, for example HATU or DCC, or by using an acid derivative such as an acyl halide or acyl imidazole. Standard methods for amide formation are commonly and widely known to those skilled in the art.

In further detail, the process comprises: (a) when L ¹ represents hydrogen and R ¹ does not represent hydrogen, reacting a compound of formula (II)

wherein LG ¹ represents a leaving group such as chloride, bromide, iodide, methanesulfonate or para-toluenesulfonate, and L, L ² , L ³ , L ⁴ , R ⁴ , R ⁵ , R ⁶ , A, W, V and X are as defined in formula (I) and (Ic), with, when Z ¹ in compounds of formula (I) is a group of formula (A), a compound of formula (III), or a suitable salt thereof such as a hydrobromide, acetate or hydrochloride salt

wherein P ¹ is hydrogen or a protective group such as terf-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 L ¹ represents hydrogen and R ¹ does not represent hydrogen, reacting a compound of formula (IV), or a suitable salt thereof

wherein L, L ² , L ³ , L ⁴ , R ⁴ , R ⁵ , R ⁶ R ⁸ A, W, V and X are as defined in formula (I), with a compound of formula (III) 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 L ¹ represents hydrogen and R ¹ represents hydrogen, reacting a compound of formula (V)

wherein LG ¹ represents a leaving group such as chloride, bromide, iodide, methanesulfonate or para-toluenesulfonate, P ² represents a protective group (e.g. tert-butylcarbonyl) and L, L ² , L ³ , L ⁴ , R ⁴ , R ⁵ , R ⁶ , R ⁸ A, W, V and X are as defined in formula (I), with a compound of formula (III), 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 L ¹ represents hydrogen and R ¹ represents hydrogen, reacting a compound of formula (Vl)

wherein L, L ² , L ³ , L ⁴ , R ⁴ , R ⁵ , R ⁶ , R ⁸ A, W, V and X are as defined in formula (I), P ² represents a protective group (e.g. tert-butylcarbonyl) with a compound of formula (III), 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 R ⁴ does not represent hydrogen, reacting a compound of formula (VII), or a suitable salt thereof

wherein L, L ¹ , L ² , L ³ , L ⁴ , R ⁴ , R ⁵ , R ⁶ , R ⁸ A ₁ W ₁ V and X are as defined in formula (I), P ³ represents hydrogen or an activating group (e.g. 3-nitrophenylsulfonyl) with a compound of formula (VIII), or a suitable salt thereof,

wherein LG ² represents a leaving group (e.g. chloride, bromide, iodide, methanesulfonate or para-toluenesulfonate) and P ¹ is as defined in compound of formula (III) in the presence of a base (e.g. when P ³ is hydrogen, potassium carbonate, triethylamine, diisopropylethylamine and, when P ³ 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,

^Ar (IX) in the presence of a base (e.g. when P ³ is hydrogen, potassium carbonate, triethylamine, diisopropylethylamine and, when P ³ is 3-nitrophenylsulfonyl, sodium hydride or lithium di-/sσ-propylamide), 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,

LG ² represents a leaving group (e.g. chloride, bromide, iodide, methanesulfonate or para-toluenesulfonate) in the presence of a base (e.g. when P ³ is hydrogen, potassium carbonate, triethylamine, diisopropylethylamine and, when P ³ 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 R ⁴ represents hydrogen, reacting a compound of formula (Xl)

wherein L, L ¹ , L ² , L ³ , L ⁴ , R ⁴ , R ⁵ , R ⁶ , A, W, V and X are as defined in formula (I), P ² represents a protective group (e.g. tert-butylcarbonyl), P ³ 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 P ³ is hydrogen, potassium carbonate, triethylamine, diisopropylethylamine and when P ³ 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 L ³ and L ⁴ each represents hydrogen, reacting a compound of formula (XII)

wherein L, L ¹ , and L ² are as defined in formula (I), P ¹ is as defined in compound of formula (III), P ³ represents a protective group (e.g. tørt-butylcarbonyl or 3- nitrophenylsulfonyl) with a compound Of formula (XIII), or a suitable salt thereof,

wherein R ⁴ , R ⁵ , R ⁶ , R ¹ , A ¹ W, V and X are as defined in formula (I), 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 L ³ and L ⁴ represents hydrogen, reacting a compound of formula (XIV)

wherein L, L ¹ , and L ² are as defined in formula (I), P ¹ is as defined in compound of formula (III), P ³ represents a protective group (e.g. ferf-butylcarbonyl or 3- nitrophenylsulfonoyl), LG ³ 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 L ¹ and L ² each represents hydrogen and R ⁴ do not represent hydrogen, reacting a compound of formula (XV), or a suitable salt thereof,

wherein L, L ³ , L ⁴ , R ¹ , R ⁴ , R ⁵ R ⁶ , A, W, V and X are as defined in formula (I) and P ¹ 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 L ¹ and L ² each represents hydrogen and R ⁴ represents hydrogen, reacting a compound of formula (XVI)

wherein L, L ³ , L ⁴ , R ⁴ , R ⁵ , R ⁶ , A, W, V and X are as defined in formula (I) and P ² 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 ⁰ 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 80°C.

Compounds of formula (II) may be prepared by reacting a compound of formula (XVII), or a suitable salt thereof,

wherein L, L ³ , L ⁴ , R ¹ , R ⁴ , R ⁵ , R ⁶ , A, W, V, and X are as defined in formula (I), with a compound of formula (XVIII)

2

L Mt ( _XV||| ) wherein L ² 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 6O ⁰ C, 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 6O ⁰ C,

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 6O ⁰ C. Compounds of formula (V) may be prepared by reacting a compound of formula (XIX)

wherein P ² , L ₁ L ³ , L ⁴ , R ⁴ , R ⁵ , R ⁶ , A, W, V 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 6O ⁰ C, 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,

/V,λ/-dimethylformamide or dimethylsulfoxide at a temperature, for example in the range from -78 to 6O ⁰ C.

Compounds of formula (VII) in which L ¹ represents hydrogen and L, L ² , L ³ , L ⁴ , R ¹ , R ⁴ , R ⁵ , R ⁶ , A, W, V, and X 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 85 ⁰ C, 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, D-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 ₁ 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 8O ⁰ C, 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, L ¹ , L ² , L ³ , L ⁴ , R ¹ , R ⁴ , R ⁵ , R ⁶ , A ₁ W, V and X are as defined in formula (VII) may be prepared by reacting a compound of formula (XX)

wherein LG ⁴ is a leaving group (e.g. hydroxyl or chloride), L, L ¹ , L ² , L ³ , L ⁴ , R ¹ , R ⁴ , R ⁵ , R ⁶ , A, W, V and X are as defined in formula (VII), with reagents such as, when LG ⁴ 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 100 ⁰ C, or when LG ⁴ 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 100 ⁰ 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), (VIII), (IX) and (X) are known in the literature or may be prepared using known techniques.

Compounds of formula (Xl) in which L ¹ 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 85 ⁰ C, 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 ₁ 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 ⁰ 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 L ⁴ is a leaving group (e.g. hydroxyl or chloride), L, L ¹ , L ² , L ³ , L ⁴ , R ⁴ , R ⁵ , R ⁶ , A, W ₁ V X and P ² are as defined in formula (Xl), with reagents such as, when LG ⁴ is hydroxyl, diphenylphosphonic azide, in a presence of an amine (e.g. triethylamine), in an organic solvent, for example, fert-butanol, tetrahydrofuran, dichloromethane, water, or a mixture thereof, at a temperature ranging from 25 to 100 ⁰ C, or when LG ⁴ is chloride, sodium azide, in an organic solvent, for example, ether, tert-butanol, tetrahydrofuran, water, or a mixture thereof, at a temperature ranging from 25 to 100 ⁰ 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 (XII) can be prepared by (a) reacting a compound of formula (XXII)

wherein P ⁵ is hydrogen or a protective group (e.g. tert-butyldimethylsilyl, tetrahydropyran) and L, L ¹ and L ² are as defined in formula (XII), with a compound of formula (VIII), (IX) or (X), or a suitable salt thereof, in the presence of a base (e.g.

potassium carbonate, triethylamine or diisopropylethylamine when P ³ is hydrogen and sodium hydride or lithium di-/so-propylamide when P ³ is 3-nitrophenylsulfonyl) in an organic solvent such as λ/,λ/-dimethylformamide, λ/-methylpyrolidinone, tetrahydrofuran, ethanol, n-butanol or dimethyl sulfoxide, at a temperature, for example, in the range from 50 to 140 ⁰ 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 (b) reacting a compound of formula (XXIII)

(XXIII) wherein P ⁶ and P ⁷ represent an acyclic or cyclic carbonyl protective group (e.g. dimetoxy or diethoxy acetal, 1 ,3-dioxolane or 1 ,3-dioxane) and L, L ¹ , L ² , and P ³ are as defined in formula (XII), with a compound of formula (VIII), (IX) or (X), or a suitable salt thereof, in the presence of a base (e.g. potassium carbonate, triethylamine or diisopropylethylamine when P ³ is hydrogen and sodium hydride or lithium di-/so- pr.opylamide when P ³ is 3-nitrophenylsulfonyl) in an organic solvent such as N, N- dimethylformamide, λ/-methylpyrolidinone, tetrahydrofuran, ethanol, n-butanol or dimethyl sulfoxide, at a temperature, for example, in the range from 50 to 140 ⁰ 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). Removal of the protective group (e.g. diluted hydrochloric acid or amberlyst-15 resin in methanol) lead to compound of formula (XII); or (c) when L ¹ represents hydrogen, reacting a compound of formula (XXIV)

wherein P ⁵ is hydrogen or a protective group (e.g. tert-butyldimethylsilyl, tetrahydropyran) and, L and L ² are as defined in formula (XlI), with a compound of

formula (III), 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 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 R ¹ represents hydrogen, reacting a compound of formula (XXV)

wherein P ⁶ and P ⁷ represent an acyclic or cyclic carbonyl protective group (e.g. dimethoxy or diethoxy acetal, 1 ,3-dioxolane or 1 ,3-dioxane) and, L and L ² are as defined in formula (XII), with a compound of formula (III), 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) can be prepared by converting compound of formula (XII), or a precursor to compound of formula (XII) 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) and (XVI) can be prepared by similar methods by reacting a compound of formula (XXVI)

wherein L, L ³ , L ⁴ , R ⁴ , R ⁵ , R ⁶ , A, W ₁ V and X are as defined in formula (XV), P ⁸ represents either R ³ as defined in compound of formula (XV) or P ² as defined in compound of formula (XVI) and LG ⁶ represent hydroxyl or a leaving group (e.g. chloride) with a compound of formula (III), or a suitable salt thereof.

When LG ⁶ 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/'-tetramethyluroniumhexafiuorophosphate (HATU), in an organic solvent, for example, /V./V-dimethylformamide or dichloromethane, at a ^j temperature, for example in the range from 0 to 6O ⁰ C,

When LG ⁶ 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 25 ⁰ C. Compounds of formula (VII), (XIX), (XX), (XXI) can be accessed through a general amination reaction of a compound of formula (XXVII)

(XXVII) wherein R ⁴ , R ⁵ , R ⁶ , A, W, V and X are as defined in formula (I), LG ⁷ represent a hydroxyl, an esther (e.g. methoxy, ethoxy), a leaving group (e.g. chloride or bromide), or an acid anhydride in the presence of appropriate reagents, (for example in the case when LG ⁷ is an etherbromide the reaction proceeds us ing a base such as sodium hydridediisopropylethylamine in a solvent such as toluene dichloromethane at a temperature ranging from 60 0 to 135 ⁰ C, with a compound of formula (XXVIII)

(XXVIII) , wherein W is as defined in compound of formula (I), when R ¹ does not represent hydrogen, P ⁹ represents R ⁴ ; when R ¹ represents hydrogen then P ⁹ represents an appropriate nitrogen protecting group, such as te/t-butoxycarbonyl,

- for compound of formula (VlI), P ⁹ represents R ⁴ , P ¹⁰ represents

wherein L, L ¹ , L ² , L ³ , L ⁴ and P ³ are as defined in compound of formula (VII); - for compound of formula (Xl), P ⁹ represents P ² , P ¹⁰ represents

wherein L, L ¹ , L ² , L ³ , L ⁴ , P ² and P ³ are as defined in compound of formula (Xl);

- for compound of formula (XIII), P ⁹ and P ¹⁰ represents represents an appropriate nitrogen protecting group, such as tert-butoxycarbonyl, followed by suitable deprotection (e.g. trifluoroacetic acid acid);

- for compound of formula (XVII), P ⁹ represents R ⁴ , P ¹⁰ represents

wherein L, L ³ , and L ⁴ are as defined in compound of formula (XVII), wherein P ¹¹ and P ¹² represent an acyclic or cyclic carbonyl protective group (e.g. dimethoxy or diethoxy acetal, 1 ,3-dioxolane or 1 ,3-dioxane), followed by suitable deprotection (e.g. diluted hydrochloric acid or amberlyst-15 resin in methanol);

- for compound of formula (XIX), P ⁹ represents P ² , P ¹⁰ represents

wherein L, L , and L are as defined in compound of formula (XIX), wherein P and

P represent an acyclic or cyclic carbonyl protective group (e.g. dimethoxy or diethoxy acetal, 1 ,3-dioxolane or 1 ,3-dioxane), followed by suitable deprotection (e.g. diluted hydrochloric acid or amberlyst-15 resin in methanol);

- for compound of formula (XX), P ⁹ represents R ⁴ , P ¹⁰ represents

wherein L, L ¹ , L ² , L ³ , and L ⁴ are as defined in compound of formula (XX), wherein P ¹⁴ represent an acid protective group (e.g. methyl, ethyl or tert-butyl), followed by suitable deprotection (e.g. lithium hydroxide or sodium hydroxide, trifluoroacetic acid, hydrochloric acid);

- for compound of formula (XXI), P ⁹ represents P ² , P ¹⁰ represents

wherein L, L ¹ , L ² , L ³ , and L ⁴ are as defined in compound of formula (XXI), wherein P ¹⁴ represent an acid protective group (e.g. methyl, ethyl or tert-butyl), followed by suitable deprotection (e.g. lithium hydroxide or sodium hydroxide, trifluoroacetic acid, hydrochloric acid);

- for compound of formula (XXVI), P ⁹ represents P ⁸ , P ¹⁰ represents

wherein L, L ³ , and L ⁴ are as defined in compound of formula (XXVI), wherein P ¹⁴ represent an acid protective group (e.g. methyl, ethyl or tert-butyl), followed by suitable deprotection (e.g. lithium hydroxide or sodium hydroxide, trifluoroacetic acid, hydrochloric acid);

When Z ¹ in compounds of formula (I) is a group of formula (B), it will be recognised by those skilled in the art that a similar set of reactions as described above may be used, employing a compound of formula (Ilia) in place of a compound of formula (III), or a suitable salt thereof such as a hydrobromide, hydrochloride or acetate salt

(Ilia)

Compounds of formula (Ilia) are known in the art, for example J. Med. Chem. 1987, 30, 1166.

Compounds of formula (XIII) and (XXVII) may be prepared using the methods illustrated in Schemes 1-17 above.

The following non-limiting Examples illustrate the invention.

General Experimental Details:

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

NMR spectra were obtained on a Varian Unity Inova 400 spectrometer with a 5mm inverse detection' triple resonance probe operating at 400 MHz or on a Bruker Avance DRX 400 spectrometer with a 5mm inverse detection triple resonance TXI probe operating at 400 MHz or on a Bruker Avance DPX 300 spectrometer with a standard 5mm dual frequency probe operating at 300 MHz. Shifts are given in ppm relative to tetramethylsiiane.

Where products were purified by column chromatography, 'flash silica' refers to silica gel for chromatography, 0.035 to 0.070 mm (220 to 440 mesh) (e.g. Fluka silica gel 60), and an applied pressure of nitrogen or air up to 10 p.s.i 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.

All compounds containing a basic centre(s), which were purified by HPLC, were obtained as the TFA salt unless otherwise stated. Preparative HPLC conditions:C18-reverse-phase column (100 x 22.5 mm i.d Genesis column with 7 μm particle size). UV detection at 230 nm.

The Liquid Chromatography Mass Spectroscopy (LC/MS) and chiral , preparative HPLC systems used: ^•

Method 1

LCMS data were obtained as follows: Waters Xterra MS C18, 5 μm (4.6 x 50 mm, flow rate 2.5 ml/min) eluting with a H ₂ O-MeCN gradient containing 0.1% v/v ammonia over 7 minutes with UV detection at 215 and 254 nm. Gradient information: 0.0 - 0.1 min: 95% H ₂ O-5% MeCN; 0.1 -5.0 min; Ramp from 95% H ₂ O-5% acetonitrile to 5% H ₂ O- 95% MeCN; 5.0 - 5.5 min: Hold at 5% H ₂ O-95% MeCN; 5.5 - 5.6 min: Hold at 5% H ₂ O-95% MeCN, flow rate increased to 3.5 ml/min; 5.6 - 6.6 min: Hold at 5% H ₂ O- 95% MeCN, flow rate 3.5 ml/min; 6.6 - 6.75 min: Return to 95% H ₂ O-5% MeCN ₁ flow rate 3.5 ml/min; 6.75 - 6.9 min: Hold at 95% H ₂ O-5% MeCN, flow rate 3.5 ml/min; 6.9 - 7.0 min: Hold at 95% H ₂ O-5% MeCN, flow rate reduced to 2.5 ml/min Mass spectra were obtained using an electrospray ionization source in either the positive (ESI ⁺ ) or negative (ESI ^" ) mode.

Method 2 LCMS data were obtained as follows: Agilent Scalar column C18, 5 μm (4.6 x 50 mm, flow rate 2.5 ml/min) eluting with a H ₂ O-MeCN gradient containing 0.1% v/v formic acid over 7 minutes with UV detection at 215 and 254 nm. Gradient information: 0.0 - 0.1 min: 95% H ₂ O-5% MeCN; 0.1 -5.0 min; Ramp from 95% H ₂ O-5% acetonitrile to 5% H ₂ O-95% MeCN; 5.0 - 5.5 min: Hold at 5% H ₂ O-95% MeCN; 5.5 - 5.6 min: Hold at 5% H ₂ O-95% MeCN, flow rate increased to 3.5 ml/min; 5.6 - 6.6 min: Hold at 5% H ₂ O-95% MeCN, flow rate 3.5 ml/min; 6.6 - 6.75 min: Return to 95% H ₂ O-5% MeCN, flow rate 3.5 ml/min; 6.75 - 6.9 min: Hold at 95% H ₂ O-5% MeCN, flow rate 3.5 ml/min; 6.9 - 7.0 min: Hold at 95% H ₂ O-5% MeCN, flow rate reduced to 2.5 ml/min Mass spectra were obtained using an electrospray ionization source in either the positive (ESI ⁺ ) or negative (ESI ^" ) mode.

Method 3

LCMS data were obtained as follows: Agilent Scalar column C18, 5 μm (4.6 x 50 mm, flow rate 1.5 ml/min) eluting with a H ₂ O-MeCN gradient containing 0.1 % v/v formic acid over 12 minutes with UV detection at 215 and 254 nm. Gradient information: 0.0 - 1.0 minutes Hold at 95% H ₂ O-5% MeCN; 1.0 - 8.0 min: Ramp from 95% H ₂ O-5% acetonitrile to 5% H ₂ O-95% MeCN; 8.0 - 9.9 min: Hold at 5% H ₂ O-95% MeCN; 9.9 - 10.0 min: Return to 95% H ₂ O-5% MeCN; 10.0 - 12.0 min: Hold at 95% H ₂ O-5% MeCN. Mass spectra were obtained using an electrospray ionization source in either the positive (ESI ⁺ ) or negative (ESI ^" ) mode.

Method 4

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

Gradient - Time flow ml/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 ionisation method - Electrospray (positive ion)

Method 5

Waters Platform LC with a C 18-reverse-phase column (30 x 4.6 mm Phenomenex Luna 3 μm particle size), elution with A: water + 0.1 % formic acid; B: acetonitrile + 0.1 % formic acid. Gradient:

Gradient - Time flow ml/min %A %B

0.00 2.0 95 5

0.50 2.0 95 5

4.50 2.0 5 95

5.50 2.0 5 95

6.00 2.0 95 5

Detection - MS, ELS, UV (200 μl split to MS with in-line UV detector) MS ionisation method - Electrospray (positive and negative ion)

Method 6

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

Gradient - Time flow ml/min %A %B

0.00 2.0 95 5

0.50 2.0 95 5

4.50 2.0 5 95

5.50 2.0 5 95

6.00 2.0 95 5

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

Method 7 Chiral compounds were separated into pure enantiomers using a 250 x 20 mm Chiralpak ® IA column packed with amylase tris(3,5-dimethylphenylcarbamate) immobilized on 5 μm silica gel. The column was eluted with desired solvents, buffered with 0.1% diethylamine. Flow rate, 18mL/min. Wavelength, 220nm.

Method 8

Waters Micromass ZQ2000 quadrupole mass spectrometer with a Higgins Clipeus 5 micron C18 100 x 3.0mm column, elution with: A: water + 0.1 % formic acid; B: acetonitrile + 0.1 % formic acid. Gradient:

Gradient - Time flow 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 (1 OOμl split to MS with in-line UV detector at 254nm) MS ionisation method - Electrospray (positive and negative ion).

Method 9

Waters ZMD quadrupole mass spectrometer with a Luna 3 micron C18(2) 30 x 4.6mm column, elution with A: water + 0.1 % formic acid; B: acetonitrile + 0.1% formic acid. Gradient:

Gradient - Time flow %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

Split - 200μl/min split to the MS with in-iine Waters 996 DAD detection.

Detection - MS, ELS, UV

MS ionisation method - Electrospray (positive and negative ion)

Method 10 Aglient LC with a C18-reverse-phase column (50 x 2.1 mm Symmetry), elution with A: acetonitrile + 0.1 % trifluoroacetic acid; B: water + 0.1% trifluoroacetic acid. Gradient:

Gradient - Time flow ml/min %A %B

0.00 1.0 5 95

8.00 1.0 95 5

8.50 1.0 95 5

8.51 1.0 100 0

9.00 1.0 100 0

9.01 1.0 5 95

Detection - MS, UV MS ionisation method - APCI (positive and negative ion)

Method 11 Chiral compounds were separated into pure enantiomers using a 50x250 Varian 'Load and Lock ¹ column packed with Chiralpak OJ (20um) silica. The column was eluted with 80:20 isohexanes:ethanol at 118ml/min flow rate monitored at 220nm. Abbreviations used in the experimental section: AIBN = 2,2'-azobis(2- methylpropionitrile); DCE = 1 ,2-dichloroethane; DCM = dichloromethane; DIPEA = di- isopropylethylamine; DMAP = dimethylaminopyridine; DMF = dimethylformamide; EtOAc = ethyl acetate; EtOH = ethanol; IMS = industrial methylated spirit; IPA = 2- propanol; MeCN - acetonitrile; MeOH = methanol; NBS = N-bromosuccinimide; RT = room temperature; Rt = retention time; SCX = strong cation ion exchange chromatography; TFA = trifluoroacetic acid; THF = tetrahydrofuran; Sat = saturated.

Example 1

[5-(Hydroxy-diphenyl-methyl)-1,3,4-oxadiazol-2-ylmethyl]- dimethyl-(3-phenoxy- propyl)-ammonium bromide

a. r5-(Hvdroxy-diphenyl-methyl)-1 ,3,4-oxadiazol-2-ylmethvπ-carbamic acid tert-butyl ester

To a solution of 5-(tert-butoxycarbonylamino-methyl)-1 ,3,4-oxadiazole-2-carboxylic acid ethyl ester (0.35 g, 1.290 mmol) in dry THF (10 ml) under an atmosphere of nitrogen at -78 ⁰ C was added in a single portion a 3M solution of phenyl magnesium bromide in diethyl ether (1.290 ml, 3.87 mmol). Upon completion of the addition the reaction mixture was allowed to warm to room temperature and stirred for 10 min. The solution was poured onto a mixture of EtOAc and 1 M HCI and the layers separated. The organic phase was washed with satd. NaHCO ₃ (aq.), water and brine, dried (MgSO ₄ ) and evaporated. The residue was purified by column chromatography using 15- 20% EtOAc/iso-hexane to give the title compound (0.21 g, 40%). LC-MS (Method 1): R _t 2.97 min, m/z 382 [MH] ⁺ . ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 7.6 (1 H, t), 7.3-7.2 (11 H, m), 4.3 (2H, d) and 1.3 (9H, s).

b. (5-Dimethylaminomethyl-1 ,3,4-oxadiazol-2-yl)-diphenyl-methanol

To an ice cold solution of [5-(hydroxy-diphenyl-methyl)-1 ,3,4-oxadiazol-2-ylmethyl]- carbamic acid tert-butyl ester (0.20 g, 0.524 mmol) in DCM (5 ml) was added TFA (1 .931 ml, 26.2 mmol). After stirring at room temperature for 1 h the solvents were evaporated and the residue dissolved in DCM (5 ml) and formaldehyde (37% in water, 0.1 18 ml, 1.573 mmol) was added. After stirring at room temperature for 20 min, sodium triacetoxyborohydride (0.556 g, 2.62 mmol) was added in a single portion. After 30 min, 1 M HCI (20 ml) was added and the reaction stirred vigourously for 30 min. The mixture was poured onto satd. NaHCO ₃ (aq.) and extracted with DCM. The combined organic phases were washed with water and brine, dried (MgSO ₄ ) and evaporated. The residue was purified by column chromatography using EtOAc as eluent to give the title compound (30 mg, 17%). LC-MS (Method 1 ): R, 2.32 min, m/z 310 [MH] ⁺ . ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 7.3-7.2 (1 1 H, m), 3.7 (2H, s) and 2.1 (6H,

S).

c. r5-(Hvdroxy-diphenyl-methyl)-1 ,3,4-oxadiazol-2-ylmethvn-dimethyl-(3-phenoxy- propyD-ammonium bromide

To a solution of (5-dimethylaminomethyl-1 ,3,4-oxadiazol-2-yl)-diphenyl-methanol (0.027 g, 0.087 mmol) in acetonitrile (1 ml) and chloroform (1 ml) was added phenoxypropyl bromide (0.184 ml, 1.047 mmol). After heating at 55 ⁰ C for 48 h the reaction mixture was cooled to room temperature and the solvents evaporated. The residue was purified by column chromatography using 0-5% MeOH/DCM to give the title compound. LC-MS (Method 3): R _t 4.04 min, m/z 444 [M-Br]+. ¹ H NMR, 400 MHz, DMSOd ₆ : δ 7.4 (5H, m), 7.3-7.2 (8H ₁ m), 6.9 (3H ₁ m), 5.1 (2H, s), 3.9 (2H, t), 3.5 (2H, m), 3.1 (6H, s) and 2.2 (2H, m).

Example 2

[3-(Hydroxy-diphenyl-methyl)-1,2,4-oxadiazol-5-ylmethyl]- dimethyl-(3-phenoxy- propyl)-ammonium bromide

a. 5-Chloromethyl-1 ,2,4-oxadiazole-3-carboxylic acid ethyl ester

To an ice cold suspension of ethyl 2-oximino-oxamate (1.17 g, 8.86 mmol) in chloroform (20 ml) was added pyridine (0.788 ml, 9.74 mmol) followed by chloroacetyl chloride (0.776 ml, 9.74 mmol) dropwise over 5 min. Upon completion of the addition the suspension was allowed to warm to room temperature and stirred for 1 h. The suspension was poured onto a mixture of DCM and water, the layers separated and the aqueous extracted with DCM (note: this resulted in a lot of precipitate that would not dissolve). The organic phase was washed with water and brine, dried (MgSO ₄ ) and evaporated. The solid residue (-900 mg) was dissolved in AcOH (8 ml) and heated at reflux for 1 h. After this time, the hot solution was poured onto a mixture of EtOAc and satd. Na ₂ CO ₃ (aq.) and the layers separated. The

organic layer was washed with further satd. Na ₂ CO ₃ (aq.) (x2), water and brine, dried (MgSO ₄ ) and evaporated to give the title compound as a brown oil (0.29 g, 17%).

b. (5-Chloromethyl-1 ,2,4-oxadiazol-3-vD-diphenyl-methanol

To a solution of δ-chloromethyl-i ^^-oxadiazole-S-carboxylic acid ethyl ester (0.29 g, 1.522 mmol) in dry THF (7 ml) under an atmosphere of nitrogen at -78 ⁰ C was added in a single portion a 3M solution of phenyl magnesium bromide in diethyl ether (1.014 ml, 3.04 mmol). As soon as the addition was complete, the dark solution was allowed to warm to room temperature. After 10 min at room temperature, the solution was poured onto a mixture of 1 M HCI and EtOAc, the layers separated and the organic layer washed with satd. NaHCO ₃ (aq.), water and brine, dried (MgSO ₄ ) and evaporated. The residue was purified by column chromatography using 10 -18% EtOAc/iso-hexane to give the title compound (0.13 g, 26%). LC-MS (Method 1): R _t 3.17 min, m/z 283 [MH-(H ₂ O)J ⁺ . ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 7.3-7.2 (1 OH, m), 7.0 (1 H, br s) and 5.1 (2H, s).

c. (5-Dimethylaminomethyl-1 ,2,4-oxadiazol-3-yl)-diphenyl-methanol A 2M solution of dimethylamine in THF (1.995 ml, 3.99 mmol) was added to (5- chloromethyl-1 ,2,4-oxadiazol-3-yl)-diphenyl-methanol (0.12 g, 0.399 mmol) and the mixture heated for 20 min at 60 ⁰ C in a CEM Discovery microwave (initial power = 300W, quick test method).The solvent was evaporated and the residue purified by column chromatography using 0-50% EtOAc/iso-hexane to give the title compound (72 mg, 58%). LC-MS (Method 1): R, 2.72 min, m/z 310 [MH] ⁺ . ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 7.3-7.2 (1 OH, m), 6.9 (1 H, s), 3.8 (2H, s) and 2.2 (6H, s).

d.r3-(Hvdroxy-diphenyl-methyl)-1 ,2,4-oxadiazol-5-ylmethyl1-dimethyl-(3-phenoxy- propyP-ammonium bromide

A solution of (5-dimethylaminomethyl-1 ,2,4-oxadiazol-3-yl)-diphenyl-methanol (0.068 g, 0.220 mmol) was reacted in a similar manner to that described in example 1 to give the title compound (39 mg, 34%). LC-MS (Method 3): R, 4.09 min, m/z 444 [M-Br] ⁺ . ¹ H NMR ₁ 400 MHz, DMSOd ₆ : δ 7.4 (4H, m), 7.3-7.2 (8H, m), 7.1 (1 H, s), 6.9 (1 H, t), 6.8 (2H, d), 5.1 (2H, s), 3.9 (2H, t), 3.6 (2H, m), 3.2 (6H, s) and 2.2 (2H, m).

Example 3

[5-(Hydroxy-diphenyl-methyl)-1,2,4-oxadiazol-3-yImethyl]- dimethyl-(3-phenoxy- propyl)-ammonium bromide

The example was prepared in a similar manner to that described in example 2, starting from 3-chIoromethyl-[1 ,2,4]oxadiazole-5-carboxylic acid ethyl ester (prepared according to patent DE1915495J. LC-MS (Method 3): R _t 4.09 min, m/z 444 [M-Br] ⁺ . DMSOd ₆ : δ 7.6 (1 H, s), 7.4 (4H, m), 7.3-7.2 (8H ₁ m), 6.9 (1 H, t), 6.8 (2H, d), 4.9 (2H, s), 3.9 (2H, t), 3.5 (2H, m), 3.2 (6H, s) and 2.2 (2H ₁ m).

Example 4

[4-(Hydroxy-diphenyI-methyl)-oxazol-2-ylmethyl]-dimethyl- (3-phenoxy-propyl)- ammonium bromide

a. (2-Methyl-oxazol-4-yl) ^' -diphenyl-methanol

To an ice-cold solution of 2-methyl-oxazole-4-carboxylic acid methyl ester (0.5 g, 3.54 mmol) under an atmosphere of nitrogen was added, dropwise, over a 20 min period a 3M solution of phenylmagnesium bromide in diethyl ether (2.36 ml, 7.09 mmol). Once the addition was complete the mixture was allowed to warm to room temperature and after 20 min the suspension was poured onto a mixture of diethyl ether and 1 M HCI. The layers were separated, the aqueous extracted with diethyl ether and the combined organic layers washed with satd. NaHCO ₃ (aq.), water and brine, dried (MgSO ₄ ) and evaporated. The residue was subjected to column chromatography (SiO ₂ , 40 g) eluting with 20% EtOAc/iso-hexane to give the title compound (0.82 g,

85%). LC-MS (Method 1 ): R, 2.84 min, m/z 266 [MH] ⁺ . ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 7.5 (1 H, s), 7.3-7.2 (8H, m), 7.2 (2H, m), 6.3 (1 H, s) and 2.4 (3H, s).

b. (2-Dimethylaminomethyl-oxazol-4-yl)-diphenyl-methanol

DCE (2.5 ml) was added to a mixture of Intermediate 1 (0.15 g, 0.565 mmol), AIBN (9.28 mg, 0.057 mmol) and λ/-bromosuccinimide (0.106 g, 0.594 mmol). The mixture was placed in a preheated oil bath at 90 ⁰ C and heated for 30 min. After this period, the hot solution was poured onto a mixture of EtOAc and satd. NaHCO ₃ (aq.) and the layers separated. The organic layer was washed with 1M HCI, water and brine, dried (MgSO ₄ ) and evaporated. The residue was dissolved in THF (2 ml) and a 2M solution dimethylamine in THF (1.413 ml, 2.83 mmol) added. After standing for 20 min the solvent was evaporated and the residue was subjected to column chromatography (SiO ₂ , 8 g) eluting with 60% EtOAc/iso-hexane to give the title compound (26 mg, 14%).LC-MS (Method 1): R, 2.62 min, m/z 309 [MH] ⁺ . ¹ H NMR ₁ 400 MHz, DMSO-d ₆ : δ 7.6 (1 H, s), 7.3 (4H, m), 7.3-7.2 (6H, m), 6.3 (1 H, s), 3.5 (2H, s) and 2.2 (6H,s).

c. r4-(Hvdroxy-diphenyl-methylVoxazol-2-ylmethyll-dimethvH3-phe noxy-propyl)-

ammonium; bromide

To a solution of (2-dimethylaminomethyl-oxazol-4-yl)-diphenyl-methanol (0.024 g, 0.078 mmol) in chloroform (0.5 ml) and CH ₃ CN (0.5 ml) was added phenoxypropyl bromide (0.147 ml, 0.934 mmol). The mixture was heated at 55 ⁰ C for 20 h. After this period the solvents were evaporated and the residue subjected to column chromatography (SiO ₂ , 8 g) eluting with 10-15% MeOH/DCM to give the title 0 compound as a white solid (15 mg, 37%). LC-MS (Method 3): R, 4.24 min, m/z 443 [M-Br] ⁺ . ¹ H NMR, 400 MHz, DMSOd ₆ : δ 7.9 (1 H, s), 7.3 (4H, m), 7.3-7.2 (8H, m), 6.9 (1 H, dt), 6.8 (2H, m), 6.5 (1 H, s), 4.8 (2H, s), 3.9 (2H, t), 3.4 (2H, m), 3.1 (6H, s) and 2.2 (2H, m).

5 The following Examples were prepared in a similar manner to that described in example 4.

Example 8

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1,2,4]oxadiazol-5- yl methyl]-dimethyl-(3- phenoxyl-propyl)-ammonium bromide, Enantiomer 1

a. 2-Dimethylamino-N-(2-oxo-2-phenyl-ethyl)-acetamide

To a solution of N.N-dimethylglycine (1.155 g, 11.2 mmol) in sieve dried DCM (80 ml_) under an atmosphere of nitrogen was added, portion wise CDI (2.03 g, 12.55 mmol). Upon addition reaction was stirred at room temperature for 2 hours. After this period phenacylamine hydrochloride (2.00 g, 11.65 mmol) was added portion wise, followed by drop wise addition of triethylamine (1.61 ml_, 11.65 mmol). The reaction was stirred under a nitrogen atmosphere at room temperature for 19 hours then washed with water (x 3), dried (MgSO ₄ ) and evaporated. The crude product was purified by chromatography eluting with 0-5% MeOH/DCM to give the title compound as a yellow oil (1.367 g). 1 H NMR, 400 MHz, CDCI ₃ : δ 7.98-8.09 (3H, m), 7.59-7.65 (1 H, m), 7.48- 7.52 (2H, m), 4.79-4.82 (2H, d), 3.03 (2H, s), 2.38 (6H, s).

b. (5-Dimethylaminomethyl-π ,2,41oxadiazole-3-yl)-phenyl-methanone 2-dimethylamino-N-(2-oxo-2-phenyl-ethyl)-acetamide (5.75 g, 26.10 mmol) was dissolved in glacial acetic acid (40 mL). A solution of sodium nitrite (2.61 g, 37.85 mmol) in deionised water (3.13 mL) was added drop wise. The resulting mixture was stirred at room temperature for approximately 210 hours adding further sodium nitrite (2.61 g, 37.85 mmol) in deionised water (3.13 mL) every 48 hours. The reaction mixture was then concentrated under reduced pressure to give a solid residue which was dissolved into DCM and washed using water, 1 M sodium hydroxide, water, dried (MgSO ₄ ) and evaporated. The crude product was purified by chromatography eluting with 0-0.5% MeOH/DCM to give the title compound as a gummy solid (1.3 g). 1 H NMR, 400 MHz, CDCI ₃ : δ 8.26-8.30 (2H, m), 7.65-7.71 (1 H, m), 7.51-7.57 (2H, m),. 3.96 (2H, s), 2.44 (6H, s).

c. Cvclohexyl-fS-dimethylaminomethyl-π ^λioxadiazole-S-vD-phenyl-methanol To an ice cold solution of (5-dimethylaminomethyl-[1 ,2,4]oxadiazole-3-yl)-phenyl- methanone (300 mg, 1.297 mmol) in anhydrous THF (10 mL) under an atmosphere of nitrogen was added, drop wise a 2M solution of cyclohexylmagnesium chloride in diethyl ether (0.973 mL, 1.946 mmol). Once the addition was complete the reaction mixture was warmed to room temperature and stirred for 18 hours. The reaction was quenched by addition of satd. NH ₄ CI solution (aq.) (2 mL). The aqueous layer was extracted using EtOAc and the combined organic layers were washed with satd. NaHCO ₃ (aq.), brine, dried (MgSO ₄ ) and evaporated. The crude product was purified by chromatography eluting with 0-0.5% MeOH/DCM to give the title compound as an oil (164 mg). The enantiomers were separated using chiral separation method 7 using 6% IPA in heptane as the eluent. Enantiomer 1 : R _t 8.50 min, enantiomer 2 : R _t 10.60 min. 1 H NMR, 400 MHz, CDCI ₃ : δ 7.60-7.65 (2H, m), 7.30-7.35 (2H, m), 7.22-7.26 (1 H, m), 3.77 (2H, s), 3,27-3.32 (1 H, m), 2.29-2.36 (7H, m), 1.60-1.78 (3H, m), 1.47- 1.54 (1 H, m), 1.09-1.42 (6H, m).

d. rS-fCyclohexyl-hydroxy-phenyl-methylHI ,2,41oxadiazol-5-yl methyli-dimethyl-O- phenoxyl-propyD-ammonium bromide, enantiomer 1

The title compound was prepared from cyclohexyl-(5-dimethylaminomethyl-

[1 ,2,4]oxadiazole-3-yl)-phenyl-methanol enantiomer 1 in a similar manner to that described in example 1. LC-MS (Method 4): R _t 8.69 min, m/z 450 [M] ⁺ . ¹ H NMR, 400

MHz, CDCI ₃ : δ 1.08-1.19 (3 H, m), 1.20-1.43 (3 H, m), 1.64-1.80 (3 H, m), 2.33-2.40

(4 H, m), 3.59 (6 H, s), 3.86 (2 H ₁ m), 4.03 (2 H, m), 5.46 (1 H, d, J = 15.07 Hz), 5.69

(1 H, d, J = 14.74 Hz), 6.83 (2 H, d, J = 8.16 Hz), 6.98 (1 H, t, J = 3.68 Hz), 7.20-

7.35(5H, m), 7.53 (2 H, d, J = 11.05 Hz).

Example 9

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1,2,4]oxadiazoI-5- yl methyl]-dimethyl-(3- phenoxyl-propyl)-ammonium bromide, enantiomer 2

The title compound was prepared from cyclohexyl-(5-dimethylaminomethyl- [1 ,2,4]oxadiazole-3-yl)-phenyl-methanol enantiomer 2 in a similar manner to that described in example 1. LC-MS (Method 4): R, 8.72min, m/z 450 [M] ⁺ . ¹ H NMR, 400 MHz, CDCI ₃ : δ 1.05-1.30 (3 H, m), 1.54-1.68 (3H, m), 1.66-1.76 (3H, m), 2.25-2.32 (4 H, m), 3.56 (6 H, s), 3.77-3.84 (2 H, m), 4.00-4.15 (2 H, m), 5.40 (1 H, d, J = 14.92 Hz), 5.62 (1 H, d, J = 14.93 Hz), 6.82 (2 H, d, J = 8.15 Hz), 6.94 (1 H, t, J = 7.36 Hz), 7.16 (1 H ₁ 1, J = 7.25 Hz), 7.22-7.28 (5 H, m), 7.47 (2 H, d, J = 7.75 Hz).

Example 10 r5-(Cvclohexyl-hvdroxy-phenyl-methylH1,2,41oxadiazol-3-yl methyli-dimethyl-Q- phenoxyl-propyD-ammortium bromide, Enantiomer 1.

a. (3-Dimethylaminomethyl-π .2,41oxadiazole-5-yl)-phenyl-methanone A solution of oxo-phenyl-acetyl chloride (3.70 g, 21.92 mmol) in acetic acid (20 mL) was added drop wise to a solution of 2-dimethylamino-acetamide oxime (2.335 g, 19.93 mmol) in acetic acid (40 mL). Upon addition the reaction was stirred at room temperature for 64 hours and then at 50 °C for 84 hours. The reaction was concentrated under reduced pressure. The residue was dissolved into DCM and washed with 2M NaOH (x2), brine, dried over (MgSO ₄ ) and evaporated. The crude product was purified by chromatography eluting with 0-0.5% MeOH/DCM to give the title compound as an oil (555mg). 1 H NMR, 400 MHz, CDCI ₃ : δ 8.38-8.43 (2H, m), 7.69-7.74 (1 H, m), 7.54-7.59 (2H, m), 3.82 (2H, s), 2.42 (6H, s).

b. Cvclohexyl-O-dimethylaminomethyl-π ^λioxadiazol-S-vπ-phenyl-methanol, The title compound was prepared from (3-dimethylaminomethyl-[1 ,2,4]oxadiazole-5- yl)-phenyl-methanone in a similar manner to that described in example 8. The enantiomers were separated using chiral separation method 7 using 6% IPA in heptane as the eluent. Enantiomer 1 : R _t 6.90 min, enantiomer 2 : R _t 6.85 min. LC-MS (Method 4): R, 6.90 min, m/z 315 [M] ⁺ . 1 H NMR, 400 MHz, CDCI ₃ : δ 1.09-1.35 (7 H, m), 1.35-1.47 (1 H, m), 1.62-1.69 (1 H, m), 1.69-1.80 (1 H, m), 2.34 (6 H, s), 3.45 (1 H, s), 3.63 (2 H, s), 7.25-7.31 (2 H, m), 7.33-7.38 (2 H, m), 7.62-7.66 (2 H, m).

c. rS-fCyclohexyl-hydroxy-phenyl-methylHI ,2,41oxadiazol-3-yl methyll-dimethyl-(3- phenoxyl-propyD-ammonium bromide, Enantiomer 1

The title compound was prepared from cyclohexyl-(3-dimethylaminomethyl- [1 ,2,4]oxadiazol-5-yl)-phenyl-methanol (enantiomer 1 ) in a similar manner to that described in Example 1. LC-MS (Method 4): R, 8.67 min, m/z 450 [M] ⁺ . ¹ H NMR, 400 MHz, CDCI ₃ : δ 1.02-1.20 (2 H, m), 1.19-1.43 (4 H, m), 1.54-1.67 (2 H, m), 1.66-1.77 (1 H, m), 1.86 (1 H, s), 2.29-2.37 (3 H, m), 3.54 (6 H, s), 3.80 (2 H, dd, J = 10.36, 5.89 Hz), 4.00-4.09 (2 H, m), 4.78 (1 H, s), 5.02 (1 H, d, J = 14.19 Hz), 5.23 (1 H, d, J = 14.17 Hz), 6.78-6.88 (2 H, m), 6.89-6.97 (1 H, m), 7.18-7.31 (5 H, m), 7.50-7.55 (2 H, m).

Example 11

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1,2,4]oxadiazol-3- yl methyl]-dimethyl-(3- phenoxyl-propyl)-ammonium bromide, Enantiomer 2.

The title compound was prepared from Cyciohexyl-(3-dimethylaminomethyl-

[1 ,2,4]oxadiazol-5-yl)-phenyl-methanol (enantiomer 2) in a similar manner to that described in Example 1. LC-MS (Method 4): R, 8.67 min, m/z 450 [M] ⁺ . ¹ H NMR, 400 MHz, CDCI ₃ : δ 0.96-1.45 (7 H, m), 1.54-1.76 (2 H, m), 2.02 (1 H, s), 2.28-2.37 (3 H, m), 3.54 (6 H, s), 3.80 (2 H, dd, J = 10.31 , 6.02 Hz), 4.00-4.09 (2 H, m), 4.85 (1 H, s), 5.02 (1 H, d, J = 14.18 Hz), 5.23 (1 H, d, J = 14.19 Hz), 6.79-6.89 (2 H, m), 6.94 (1 H, t, J = 7.36 Hz), 7.2-7.35 (5H), 7.50-7.56 (2 H, m).

Example 12, 13

[5-(Cvclopentyl-hvdroxy-phenyl-methylH1 ,2,41oxadiazol-3-yl methvπ-dimethyl- (3-phenoxyl-propyl)-ammonium bromide, enantiomer 1 and r5-(Cyclopentyl- hvdroxy-phenyl-methylH1,2,41oxadiazol-3-yl methvπ-dimethyl-(3-phenoxyl- propyP-ammonium bromide, enantiomer 2

a. Cvclopentyl-O-dimethylaminomethyl-π ^λloxadiazole-S-vD-phenyl-methanol, The title compound was prepared from (3-dimethylaminomethyl-[1 ,2,4]oxadiazole-5- yl)-phenyl-methanone in a similar manner to that described in example 10. The enantiomers were separated using chiral separation method 4 using 2.5% EtOH in heptane as the eluent. Enantiomer 1 : R _t 10.72 min, enantiomer 2 : R _t 12.03 min. LC- MS (Method 4): R, 6.40 min, m/z 302 [M] ⁺ . 1 H NMR, 400 MHz, CDCI ₃ : δ 1.31 -1.72 (8 H, m), 2.31 (6 H, s), 2.96-3.08 (1 H, m), 3.55 (1 H, d, J = 14.17 Hz), 3.60 (2 H, s), 7.24-7.29 (1 H, m), 7.32-7.37 (2 H, m), 7.61-7.65 (2 H, m).

b. ^-(Cyclopentyl-hydroxy-phenyl-methylHI ,2,41oxadiazol-3-yl methyl]-dimethyl-(3- phenoxyl-propyO-ammonium bromide, enantiomer 1

The title compound, Example 12, was prepared from cyclopentyl-(3- dimethylaminomethyl-[1 ,2,4]oxadiazole-5-yl)-phenyl-methanol (enantiomer 1) in a similar manner to that described in Example 1. LC-MS (Method 4): R _t 8.39 min, m/z 436 [M] ⁺ . ¹ H NMR, 400 MHz, CDCI ₃ : δ 1.25-1.76 (8 H, m), 2.31-2.38 (2 H, m), 2.89- 3.03 (1 H, m), 3.53 (6 H, s), 3.82 (2 H, dd, J = 10.15, 6.18 Hz), 4.06 (2 H, t, J = 5.54 Hz), 4.94 (2 H, d, J = 13.42 Hz), 5.20-5.29 (1 H, m), 6.81-6.86 (2 H, m), 6.95 (1 H, t, J = 7.36 Hz), 7.20-7.33 (5 H, m), 7.48-7.53 (2 H, m).

[δ-fCvclopentyl-hvdroxy-phenyl-methvD-fi ^λioxadiazol-S-yl-methvπ-dimethyl-O- phenoxyl-propyD-ammonium bromide, enantiomer 2 The title compound, Example 13, was prepared from cyclopentyl-(3- dimethylaminomethyl-[1 ,2,4]oxadiazole-5-yl)-phenyl-methanol (enantiomer 2) in a similar manner to that described in Example 1. LC-MS (Method 4): R _t 8.34 min, m/z 436 [M] ⁺ . ¹ H NMR, 400 MHz CDCI ₃ : δ 1.24-1.77 (8 H, m), 2.31-2.37 (2 H, m), 2.90- 3.02 (1 H, m), 3.53 (6 H, s), 3.82 (2 H, dd, J = 10.09, 6.27 Hz), 4.03-4.11 (2 H, m),

4.88-4.99 (2 H, m), 5.25 (1 H, d, J = 14.11 Hz) ₁ 6.82-6.88 (2 H, m), 6.94 (1 H, t, J = 7.35 Hz), 7.18-7.33 (5 H, m), 7.51 (2 H, d, J = 7.67 Hz).

Example 14 [5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1 ,3,4]oxadiazol-2-ylmethyl]-dimethyl-(3- phenoxy-propyl)-ammonium bromide, enantiomer 1

a. Dimethylamino-acetic acid N'-(2-oxo-2-phenyl-acetyl)-hydrazide To a flask containing phenyl glyoxylic acid (4.23 g, 28 mmol), cooled in an ice-bath, was added oxalyl chloride (7 mL). The reaction was initiated by the addition of 1 drop of DMF. The mixture was gradually allowed to warm to room temperature over 2 hours after which the excess oxalyl chloride was evaporated off under reduced pressure. The crude acid chloride was dissolved in THF (10 mL) and added dropwise to a cooled (0 ⁰ C) solution of dimethylamino-acetic acid hydrazide (2.75 g, 24 mmol) and triethylamine (4.9 mL, 35 mmol) in THF (20 mL). This mixture was allowed to warm to room temperature and stirred for 2 hours. Water (50 mL) was added and the mixture extracted with EtOAc (3 x 50 mL). The combined organics were dried over Na ₂ SO ₄ , filtered and evaporated to a yellow solid. Purification on the CombiFlash Companion ® system over silica using a 0 - 15 % MeOH/DCM as eluent gave the title compound as a yellow foam (1.7 g, 24 %). LC-MS (Method 5): R, 0.32 min, m/z 250 [M+Hf.

b. (5-Dimethylaminomethyl-H ,3,41oxadiazol-2-yl)-phenyl-methanone

A solution of dimethylamino-acetic acid N'-(2-oxo-2-phenyl-acetyl)-hydrazide (1.7 g, 6.8 mmol) was formed in DCM (30 mL). Triethylamine (2.4 mL, 17 mmol) and tosyl chloride (1.3 g, 6.8 mmol) were added and the mixture stirred at room temperature for 2 hours. NaHCO ₃ (sat. aq. 10 mL) was added and the organics isolated through a phase separation cartridge then evaporated. Purification on the CombiFlash

Companion ® system over silica using a 0 - 10 % gradient of MeOH/ DCM as eluent

gave the title compound as an orange oil (1.0 g, 64 %). LC-MS (Method 5): R _t 1.51 min, m/z 273 [M+H+MeCN]+, m/z 232 [M+H]+.

c. Cvclohexyl-fδ-dimethylaminomethyl-H .SλIoxadiazol^-vO-phenyl-methanol, enantiomer 1

Cyclohexyl magnesium chloride was reacted with (5-dimethylaminomethyl-

[1 ,3,4]oxadiazol-2-yl)-phenyl-methanone using a method analogous to that in example

8 to give the racemic title compound as a colourless oil.

The two enantiomers were isolated following preparative chiral HPLC of the racemate (method 7; 10 % ethanol / 90 % heptane / 0.1 % diethylamine; R _t 13 min). Enantiomer 1 : LC-MS (method 4): R, 5.52 min, m/z 315 [M+H]+. ¹ H NMR, 400 MHz, CDCI ₃ : δ 1.07-1.24 (3 H, m), 1.23-1.44 (4 H, m), 1.63-1.71 (3 H, m), 1.72-1.82 (1 H, m), 2.31 (6 H, s), 3.37 (1 H ₁ s), 3.76 (2 H, s), 7.27-7.31 (1 H, m), 7.32-7.38 (2 H, m), 7.56-7.60 (2 H, m). Enantiomer 2: (method 7; 10 % ethanol / 90 % heptane / 0.1 % diethylamine; Rt 15 min). LC-MS (method 4): R, 5.62 min, m/z 315 [M+H]+. ¹ H NMR, 400 MHz, CDCI ₃ : δ 1.13-1.19 (3 H, m), 1.25-1.43 (4 H, m), 1.62-1.72 (3 H, m), 1.72-1.82 (1 H, m), 2.31 (6 H, s), 3.36 (1 H, s), 3.76 (2 H, s), 7.27-7.31 (1 H, m), 7.32-7.40 (2 H, m), 7.55-7.60 (2 H, m).

d. r5-(Cvclohexyl-hvdroxy-phenyl-methyl)-π ,3,41oxadiazoi-2-ylmethvn-dimethyl-(3- phenoxy-propyD-ammonium bromide, enantiomer 1

The title compound was formed by the reaction of cyclohexyl-(5-dimethylaminomethyl- [1 ,3,4]oxadiazol-2-yl)-phenyl-methanol, enantiomer 1 with 3-bromo-1-phenoxypropane using an analogous method to that in example 1 : LC-MS (method 4): R, 8.34 min, m/z 450 [M]+; ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 0.91-1.36 (6 H, m), 1.60 (2 H ₁ 1, J = 15.02 Hz) ₁ 1.70 (2 H, d, J = 9.97 Hz) ₁ 2.20-2.32 (3 H ₁ m), 3.18 (6 H ₁ s), 3.53 (2 H ₁ dd, J = 10.64, 5.79 Hz) ₁ 4.00 (2 H ₁ 1, J = 5.86 Hz) ₁ 5.05 (2 H ₁ s), 6.45 (1 H ₁ s), 6.91 -6.99 (3 H ₁

170

87 m), 7.23-7.37 (5 H, m), 7.44-7.49 (2 H, m).

Example 15

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1,3,4]oxadiazol-2- ylmethyl]-dimethyl-(3- 5 phenoxy-propyl)-ammonium bromide, enantiomer 2

The title compound, Example 15, was formed by the reaction of cyclohexyl-(5- dimethylaminomethyl-[1 ,3,4]oxadiazol-2-yl)-phenyl-methanol, enantiomer 2 with 3- bromo-1 -phenoxypropane using an analogous method to that in example 1. LC-MS 0 (method 4): R, 8.30 min, m/z 450 [M]+; ¹ H NMR, 400 MHz DMSOd ₆ : δ 0.86-1.35 (6 H, m), 1.60 (2 H, t, J = 14.96 Hz), 1.70 (2 H, d, J = 10.02 Hz), 2.20-2.31 (3 H, m), 3.19 (6 H, s), 3.53 (2 H, dd, J = 10.65, 5.81 Hz), 4.01 (2 H, t, J = 5.87 Hz), 5.06 (2 H, s), 6.45 (1 H, s), 6.91 -6.99 (3 H, m), 7.23-7.37 (5 H, m), 7.44-7.49 (2 H, m)..

5 The following compounds were made using methods analogous to those in examples 14, 15 and 1. Enantiomers of cyclopentyHδ-dimethylaminomethyl-i ,3,4]oxadiazol-2- yl)-phenyl-methanol were separated using chiral separation method 7 using 5% IPA in heptane as the eluent: enantiomer 1 (R _t 11.9 min), enantiomer 2 (R _t 12.9 min).

Example 18

[5-(Hydroxy-di-thiophen-2-yl-methyl)-[1,3,4]oxadiazol-2-y lmethyl]-dimethyl-(3- phenoxy-propyl)-ammonium bromide:

a. Dimethylamino-acetic acid N'-(2-oxo-2-thiophen-2-yl-acetyl)-hvdrazide The title compound was synthesised from 2-thiopheneglyoxylic acid and dimethylamino-acetic acid hydrazide using a method analogous to that in example 18a: LC-MS (Method 5): R _t 0.55 min, m/z 256 [M+H]+.

b. (5-Dimethylaminomethyl-π ,3,41oxadiazol-2-yl)-thiophen-2-yl-methanone The title compound was synthesised from dimethylamino-acetic acid N'-(2-oxo-2- thiophen-2-yl-acetyl)-hydrazide using a method analogous to that in example 18b: LC- MS (Method 6): R _t 1.60 min, m/z 238 [M+H]+.

c. (5-Dimethylaminomethyl-π ,3,41oxadiazol-2-yl)-di-thiophen-2-yl-methanol 2-Thienyl magnesium bromide was reacted with (5-dimethylaminomethyl-

[1 ,3,4]oxadiazol-2-yl)-thiophen-2-yl-methanone using a method analogous to that in example 8 to give the title compound as a pale green crystalline solid: LC-MS (method 4): R ₁ 5.40 min, m/z 321 [M+H]+; ¹ H NMR, 400 MHz, CDCI ₃ : δ 2.29 (6 H, s), 3.77 (2 H, s), 4.84 (1 H, s), 6.99 (2 H, dd, J = 5.09, 3.67 Hz), 7.09 (2 H, dd, J = 3.66, 1 .25 Hz), 7.34 (2 H, dd, J = 5.09, 1.25 Hz). d. f5-(Hvdroxy-di-thiophen-2-yl-methyl)-f1 ,3,4loxadiazol-2-ylmethvπ-dimethyl-(3- phenoxy-propyD-ammonium bromide:

The title compound was formed by the reaction of (5-dimethylaminomethyl- [1 ,3,4]oxadiazol-2-yl)-di-thiophen-2-yl-methanol with 3-bromo-1 -phenoxypropane using an analogous method to that in example 1 : LC-MS (method 4): R, 7.50 min, m/z 456 [M]+; ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 2.20-2.33 (2 H, m), 3.21 (6 H, s), 3.53-3.60

(2 H, m), 4.01 (2 H ₁ 1, J = 5.88 Hz), 5.11 (2 H, s), 6.90-7.01 (5 H, m), 7.12 (2 H ₁ dd, J = 3.64, 1.28 Hz), 7.28-7.34 (2 H, m), 7.54 (2 H, dd, J = 5.08, 1.27 Hz), 8.12 (1 H, s).

Example 19 [δ^Cyclohexyl-hydroxy-phenyl-methylJ-thiazol^-ylmethylJ-dim ethyl^S-phenoxy- propyl)-ammonium bromide, Enantiomer 1:

a. Dimethyl-thiazol-2-ylmethyl-amine A solution of dimethylamine (10 ml_ of 2 M in THF, 20 mmol) was added to thiazole 2- carboxaldehyde (1.13 g, 10 mmol). The resulting solution was cooled in an ice bath and sodium triacetoxyboro hydride (4.24 g, 20 mmol) was added portionwise. THF (10 mL) was added to aid stirring and the mixture was stirred at room temperature over night. The mixture was partitioned between ethyl acetate (50 mL) and aqueous NaHCO ₃ (sat. 20 mL). The aqueous phase was extracted with DCM (2 x 30 mL). The combined organics were dried over sodium sulphate, filtered and evaporated to a brown oil. Purification on the CombiFlash Companion ® system over silica using a 0 - 10 % MeOH/DCM as eluent then Kugelrohr distillation gave the title compound as a colourless oil (130 mg, 9 %): bpt. 75 ⁰ C / 2 mmHg; ¹ H NMR, 400 MHz, CDCI ₃ : δ 2.37 (6 H, s), 3.83 (3 H, s), 7.31 (1 H, d, J = 3), 7.71 (1 H, d, J = 3).

b. Cvclohexyl-(2-dimethylaminomethyl-thiazol-5-yl)-phenyl-metha nol:

A solution of dimethyl-thiazol-2-ylmethyl-amine (130 mg, 0.9 mmol) was formed in dry THF (3 mL) and cooled to -78 ⁰ C. A solution of n-butyl lithium (0.69 mL of 1.6 M in THF, 1.1 mmol) was added dropwise. The mixture was stirred at -78 °C for 30 minutes and then a solution of cyclohexyl phenyl ketone (207 mg, 1.1 mmol) in dry THF (2 mL) was added dropwise. After a further 10 minutes at -78 ⁰ C the mixture was allowed to warm to room temperature over 2 hours. The reaction was quenched by the addition of aqueous NH ₄ CI (sat. 2 mL) then partitioned between EtOAc (10 mL)

and water (10 ml_). The aqueous phase was extracted with EtOAc (10 mL) and the combined organics were dried over sodium sulphate, filtered and evaporated. Purification on the Combi Flash Companion ® system over silica using a 0 - 100 % EtOAc/DCM gives the racemic title compound as a pale yellow oil (230 mg, 77 %). The enantiomers were separated by preparative chiral HPLC of the racemate. Enantiomer 1 : (method 7; 5 % /so-propanol / 95 % heptane / 0.1 % diethylamine; R _t 13 min): LC-MS (method 4): R, 6.89 min, m/z 330 [M+H]+; ¹ H NMR, 400 MHz, CHCI ₃ : δ 0.98-1.15 (3 H ₁ m), 1.15-1.36 (2 H, m), 1.45 (1 H, d, J = 12.93 Hz), 1.67-1.88 (4 H, m), 2.20 (1 H, tt, J = 11.85, 2.87 Hz), 2.33 (6 H, s), 2.39 (1 H, s), 3.69 (2 H, s), 7.23 (1 H, m), 7.29-7.34 (2 H, m), 7.44-7.54 (3 H, m). Enantiomer 2: (method 7; 5 % /so- propanol / 95 % heptane / 0.1 % diethylamine; R _t 15.5 min): LC-MS (method 4): R _t 6.78 min, m/z 330 [M+H]+; ¹ H NMR, 400 MHz, CHCI ₃ : δ 0.96-1.39 (6 H, m), 1.45 (1 H, d, J = 13.10 Hz), 1.69 (2 H, t, J = 15.12 Hz), 1.81 (2 H, t, J = 11.13 Hz), 2.15-2.24 (1 H, m), 2.33 (6 H, s), 3.69 (2 H, s), 7.21-7.26 (1 H, m), 7.33 (2 H, t, J = 7.64 Hz), 7.43- 7.56 (3 H, m).

c. fS-fCvclohexyl-hvdroxy-phenyl-methvD-thiazol^-ylmethyll-dime thyl-O-phenoxy- propyD-ammonium bromide, Enantiomer 1 : The title compound was formed by the reaction of cyclohexyl-(2-dimethylaminomethyl- thiazol-5-yi)-phenyl-methanol, Enantiomer 1 with 3-bromo-1-phenoxypropane using an analogous method to that in example 1 : LC-MS (method 4): R _t 8.22 min, m/z 465 [M]+; ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 0.97-1.28 (6 H, m), 1.49-1.72 (4 H, m), 2.19- 2.39 (3 H, m), 3.12 (6 H, s), 3.48-3.55 (2 H, m), 4.03 (2 H, t, J = 5.86 Hz), 4.89 (2 H, s), 6.00 (1 H, s), 6.89-6.98 (3 H, m), 7.19-7.38 (5 H, m), 7.53-7.58 (2 H, m), 8.00 (1 H, s).

Example 20

[δ^Cyclohexyl-hydroxy-phenyl-methyO-thiazol^-ylmethyll-d imethyl^S-phenoxy- propyl)-ammonium bromide, Enantiomer 2:

The title compound was formed by the reaction of cyclohexyl-(2-dimethylaminomethyl- thiazol-5-yl)-phenyl-methanol, Enantiomer 2 with 3-bromo-1-phenoxypropane using an analogous method to that in example 1 : LC-MS (method 4): R _t 8.19 min, m/z 465 [M]+; ¹ H NMR, 400 MHz, DMSOd ₆ : δ 1.00-1.30 (6 H, m), 1.53-1.73 (4 H, m), 2.20- 2.38 (3 H, m), 3.12 (6 H, s), 3.48-3.55 (2 H, m), 4.03 (2 H, t, J = 5.86 Hz), 4.89 (2 H, s), 5.99 (1 H, s), 6.89-6.98 (3 H, m), 7.21 (1 H, t, J = 7.31 Hz), 7.26-7.36 (4 H, m), 7.53-7.58 (2 H, m), 8.00 (1 H, s).

Example 21

[3-Hydroxy-diphenyl-methyl)-isoxazol-5-ylmethyl]-dimethyl -(3- phenoxypropyl)ammonium bromide

The title compound was formed by the reaction of 5-methyl-isoxazole-3-carboxylic acid methyl ester using an analogous method to that in example 4: LC-MS (method 3): R _t 2.34 min, m/z 443 [M]+; ¹ H NMR, 400 MHz, DMSOd ₆ : δ 2.20-2.30 (2H), 3.10 (6H, s), 3.45-3.55 (2H, m), 3.95-4.05 (2H, m), 4.87 (2H, s), 6.85-6.95 (5H, m), 7.20- 7.35 (12H ₁ m).

Example 22

[5-(Hydroxy-diphenyl-methyl)-2H-pyrazol-3-ylmethyl]dimeth yl-(3-phenoxypropyl) ammonium bromide

a. S-Dimethylaminomethyl-I H-pyrazole-S-carboxylic acid ethyl ester 1 -Dimethylamino-2-propyne (0.405mL, 3.8OmL) was added to a solution of ethyl diazoacetate (0.4ml_, 3.8mmol) in toluene (2mL) and the reaction heated to 14O ⁰ C for 15 minutes in a CEM microwave reactor, Concentration in vacuo and purification by chromatography on SiO ₂ using EtOAc then 8-20% MeOH/DCM as eluent gave the title compound as an oil. LC-MS (method 1): R ₁ 1.59 min, m/z 198 [MH]+.

b. [5-(Hydroxy-diphenyl-methyl')-2H-pyrazol-3-ylmethylldimethyl -(3-phenoxypropyπ ammonium bromide

The title compound was formed by the reaction of 5-dimethylaminomethyl-1 H- pyrazole-3-carboxylic acid ethyl ester using an analogous method to that in example 1 : LC-MS (method 3): R ₁ 2.22 min, m/z 442 [M]+; ¹ H NMR, 400 MHz, DMSO-d ₆ : 2.20- 2.30 (2H, m), 3.03 (6H, s), 3.30-3.50 (2H, m), 3.90-4.10 (2H, m), 4.50 (2H, s), 6.04 (1H, s), 6.65 (1H, s), 6.80-7.00 (3H, m), 7.20-7.40 (12H, m), 13.2 (1H, s).

Example 23

[5-(Hydroxy-diphenyl-methyl) -1 H-[1 ,2,4]triazol-3-ylmethyl]-dimethyl-(3- phenoxypropyl)ammonium bromide

The title compound was formed by the reaction of 5-(tert-butoxycarbonylamino- methyl)

-2H-1 ,2,4-triazole-3-carboxylic acid ethyl ester using an analogous method to that in example 1 : LC-MS (method 3): R, 2.17 min, m/z 443 [M-Br]+; ¹ H NMR, 400 MHz,

DMSO-d ₆ : 2.16-2.28 (2H, m), 3.08 (6H, s), 3.30-3.40 (2H, m), 3.88-3.96 (2H, m), 4.60

(2H, s), 6.80-6.85 (2H, m), 6.88- 6.95 (1H, m), 7.10-7.35 (13H ₁ m), 14.5 (1 H, br s)..

Example 24

[2-(Hydroxy-diphenyl-methyl)-3-methyl-3H-imidazol-4-ylmet hyl]-dimethyl-(3- phenoxy-propyl)ammonium bromide

a. dimethyl-(3-methyl-3H-imidazol-4-ylmethvπ-amine

Thionyl chloride (3.88mL, 53.5mmol) was added dropwise to 3-methyl-3H-imidazol-4- yl)-methanol (o.3g, 2.68mmol). The reaction was sonicated briefly, treated with DMF (1 drop) and allowed to stir at ambident temperature for 1 h. The volatiles were removed and the residue treated with diethyl ether. The diethyl ether was decanted and the procedure repeated 3 times. The semi-solid residue was treated with dimethylamine in THF (6.69mL, 13.38mmol) followed DMF (1 mL). The reaction was heated in a CEM microwave reactor at 8O ⁰ C for 30 minutes, treated with acetic acid (1 ml_) and purified by SCX cartridge to give the title compound (0.18g, 1.22mmol). LC-MS (method 1): R, 2.9 min, m/z 140 [MH]+.

b. r2-(Hvdroxy-diphenyl-methvπ-3-methyl-3H-imidazol-4-ylmethyl l-dimethyl-(3- phenoxy-propyOammonium bromide

The title compound was formed by the reaction of dimethy!-(3-methyl-3H-imidazol-4- ylmethyl)-amine using analogous methods to that described in example 19: LC-MS (method 3): R _t 2.00 min, m/z 456 [M]+; ¹ H NMR, 400 MHz, DMSOd ₆ : 2.20-2.30 (2H, m), 2.95 (6H, s), 3.38 (3H, s), 3.40-3.50 (2H, m), 4.00-4.10 (2H, m), 4.60 (2H, s), 6.80-

7.00 (4H, m), 7.10-7.40 (13H, m).

Example 25 ^-(Cyclohexyl-hydroxy-phenyl-methylJ-lsoxazoI-S-ylmethyll-di methyl^S- phenoxy-propyl)-ammonium bromide, Enantiomer 1

a. (S-Chloromethyl-isoxazol-S-yD-cyclohexyl-phenyl-methanol To a solution of (5-(chloromethyl)isoxazol-3-yl)(phenyl)methanone (7 g, 31 .58 mmol) (Med. Chem. Res 10 (9), 615-633 (2001)) at -78°C under nitrogen was added cyclohexyl magnesium chloride 2M in diethyl ether (17.37 ml_, 34.74 mmol) dropwise over 10 minutes. The reaction was stirred for 2 hours and then 1 M HCI (aqueous, 200 ml_) added. The mixture was allowed to warm to room temperature and then diluted with diethyl ether (200 ml_). The organic layer was separated and the aqueous re- extracted with diethyl ether (2 x 100 ml_). The combined organic extracts were dried over magnesium sulfate and concentrated under vacuum. Purification of the resultant crude oil was carried out by column chromatography on silica eluting with ethyl acetate / isohexane (10/90) to yield (5-(chloromethyl)isoxazol-3-yl)-cyclohexyl-phenyl- methanol as an off white solid (5.60 g, 58.0 %). The enantiomers were separated by preparative chiral HPLC of the racemate. Enantiomer 1 : (method 11 ; 20 % ethanol / 80 % isohexane ; R ₁ 6.41 min): ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 0.94 - 1.44 (6 H, m), 1.51 - 1.78 (4H, m), 2.15 - 2.26 (1 H, m), 4.89 (2H, s), 5.80 (1 H, s), 6.50 (1 H, s), 7.20 (1 H, tt, J = 7.3, 1.4 Hz), 7.31 (2H, t, J = 7.7 Hz), 7.45 - 7.49 (2H, m). Enantiomer 2: (method 11 ; 20 % ethanol / 80 % isohexane ; R _t 10.1 min): ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 0.96 - 1.33 (6H, m), 1.53 - 1.72 (4H, m), 2.14 - 2.25 (1 H, m), 4.89 (2H, s), 5.80 (1H, s), 6.50 (1 H, s), 7.20 (1H, dt, J = 14.5, 1.3 Hz), 7.31 (2H, t, J = 8.0 Hz), 7.43 - 7.52 (2H, m).

b. Cvclohexyl-fδ-dimethylaminomethyl-isoxazol-S-vO-phenyl-meth anol, Enantiomer 1 : To (δ-chloromethyl-isoxazol-S-yO-cyclohexyl-phenyl-methanol (400 mg, 1 .31 mmol) was added 2M dimethylamine solution in tetrahydrofuran (5 ml_, 10.00 mmol). The mixture was stirred over 48 hours at room temperature and then poured into water and extracted with EtOAc (3 x 20 mL). The combined organic extracts were dried over

MgSO ₄ and evaporated to give an oil which was further purified by chromatography on SiO ₂ eluting with EtOAc / DCM (25:75) and then methanol / DCM (10:90) to give cyclohexyl-(5-dimethylaminomethyl-isoxazol-3-yl)-phenyl-meth anol, Enantiomer 1 (350 mg, 85 %) as a colourless oil. LC-MS (method 10): R _t 3.14 min, m/z 315 [M+H]+. .

c. [S-tCvclohexyl-hvdroxy-phenyl-methyπ-isoxazol-δ-ylmethyli- dimethyl-O-phenoxy- propylVammonium bromide, Enantiomer 1 :

The title compound was formed by the reaction of cyclohexyl-(5-dimethylaminomethyl- isoxazol-3-yl)-phenyl-methanol, Enantiomer 1 : with 3-bromo-1 -phenoxypropane using an analogous method to that in example 1 : LC-MS (method 10): Rt 4.19 min, m/z 499 [M]+; ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 0.98 - 1.30 (6H, m), 1.54 - 1.73 (4H, m), 2.18 - 2.30 (3H, m), 3.10 (6H, s), 3.42 - 3.51 (2H, m), 4.04 (2H, t, J = 5.9 Hz), 4.85 (2H, s), 5.90 (1 H, s), 6.85 (1 H, d, J = 1.0 Hz), 6.90 - 7.00 (3H, m), 7.19 - 7.25 (1 H, m), 7.26 - 7.36 (4H, m), 7.50 (2H, d, J = 7.4 Hz).

Example 26

^-(Cyclohexyl-hydroxy-phenyl-methyO-isoxazol-S-ylmethyll- dimethyl^S- phenoxy-propyl)-ammonium bromide, Enantiomer 2

a. Cvclohexyl-(5-dimethylaminomethyl-isoxazol-3-yl)-phenyl-meth anol, Enantiomer 2:

The title compound was formed by the reaction of (5-chloromethyl-isoxazol-3-yl)- cyclohexyl-phenyl-methanol, Enantiomer 2 with dimethylamine solution using an analogous method to that in example 25: LC-MS (method 10): Rt 3.14 min, m/z 315 [M+H]+. ¹ H NMR, 400 MHz, CHCI ₃ : δ 1.04 - 1.39 (6H, m), 1.55 - 1.82 (4H, m), 2.21 - 2.31 (7H, m), 2.95 (1 H, s), 3.56 (2H, s), 6.17 (1 H, s), 7.20 - 7.25 (1 H, m), 7.29 - 7.36 (2H, m), 7.48 - 7.60 (2H, m).

b. ^-(CvclohexyI-hvdroxy-phenvI-methvD-isoxazol-S-ylmethyll-dim ethyl-O-phenoxy- propyD-ammonium bromide, Enantiomer 2:

The title compound was formed by the reaction of cyclohexyl-(5-dimethylaminomethyl- isoxazol-3-yl)-phenyl-methanol, Enantiomer 2: with 3-bromo-1 -phenoxypropane using an analogous method to that in example 1 : LC-MS (method 10): Rt 4.19 min, m/z 499 [M]+; ¹ H NMR, 400 MHz, DMSOd ₆ : δ 1.02 - 1.32 (6H, m), 1.54 - 1.73 (4H, m), 2.17 - 2.30 (3H, m), 3.11 (6H, s), 3.43 - 3.51 (2H, m), 4.04 (2H, t, J = 5.8 Hz), 4.87 (2H, s), 5.91 (1 H, s), 6.86 (1 H ₁ s), 6.90 - 7.00 (3H, m), 7.19 - 7.25 (1 H, m), 7.27 - 7.36 (4H, m), 7.50 (2H, d, J = 7.0 Hz).

Example 27

^-(Cyclohexyl-hydroxy-phenyl-methylJ-isoxazol-S-ylmethyll -^S^-dichloro- benzyloxy)-ethyl]-dimethyl-ammonium bromide

The title compound, Example 27, was formed by the reaction of cyclohexyl-(5- dimethylaminomethyl-isoxazol-3-yl)-phenyl-methanol, Enantiomer 2: with 4-(2-bromo- ethoxymethyl)-1 ,2-dichloro-benzene using an analogous method to that in example 1. LC-MS (method 10): Rt 4.88 min, m/z 517 [M]+; ¹ H NMR, 400 MHz, DMSOd ₆ : δ 0.96 - 1.30 (6H, m), 1.51 - 1.74 (4H, m), 2.16 - 2.26 (1 H, m), 3.12 (6H, s), 3.61 - 3.66 (2H, m), 3.95 - 4.00 (2H ₁ m), 4.56 (2H, s), 4.91 (2H ₁ s), 5.89 (1 H, S) ₁ 6.83 (1 H, s), 7.22 (1 H, t, J = 7.4 Hz) ₁ 7.29 - 7.38 (3H, m), 7.47 - 7.51 (2H ₁ m), 7.61 - 7.65 (2H, m).

Example 28 [3-(Cyclohexyl-hydroxy-phenyl-methyl)-isoxazol-5-ylmethyl]-[ 3-(3,4-dichloro- phenoxy)-propyl]-dimethyl-ammonium bromide

The title compound, Example 28, was formed by the reaction of cyclohexyl-(5- dimethylaminomethyl-isoxazol-3-yl)-phenyl-methanol, Enantiomer 2: with 4-(3-bromo- propoxy)-1 ,2-dichloro-benzene using an analogous method to that in example 1. LC- MS (method 10): Rt 4.35 min, m/z 517 [M]+; ¹ H NMR, 400 MHz, DMSOd ₆ : δ 0.97 - 1.-32 (6H, m), 1.54 - 1.71 (4H, m), 2.18 - 2.30 (3H, m), 3.09 (6H, s), 3.40 - 3.47 (2H ₁ m), 4.09 (2H, t, J = 5.8 Hz), 4.85 (2H, s), 5.91 (1 H ₁ s), 6.85 (1 H, s), 6.96 (1 H, dd, J = 8.8, 3.0 Hz), 7.19 - 7.26 (2H, m), 7.30 - 7.35 (2H, m), 7.47 - 7.52 (2H, m), 7.55 (1 H, d, J = 9.0 Hz).

Example 29

[S-tCyclohexyl-hydroxy-phenyl-methylJ-isoxazol-S-ylmethyl l-dimethyl^- phenethyloxy-ethyl)-ammonium bromide

The title compound, Example 29, was formed by the reaction of cyclohexyl-(5- dimethylaminomethyl-isoxazol-3-yl)-phenyl-methanol, Enantiomer 2: with [2-(2-bromo- ethoxy)-ethyi]-benzene using an analogous method to that in example 1. LC-MS (method 10): Rt 4.51 min, m/z 463 [M]+; ¹ H NMR, 400 MHz, DMSOd ₆ : δ 0.98 - 1.30 (6H, m), 1.55 - 1.74 (4H ₁ m), 2.16 - 2.26 (1 H, m), 2.83 (2H, t, J = 6.7 Hz), 2.99 (6H, s), 3.49 - 3.56 (2H, m), 3.69 (2H, t, J = 6.7 Hz), 3.83 - 3.90 (2H, m), 4.73 (2H, s), 5.89 (1 H, s), 6.77 (1 H, s), 7.12 - 7.18 (1 H, m), 7.20 - 7.26 (5H, m), 7.31 - 7.37 (2H, m), 7.48 - 7.52 (2H, m).

Example 30 ^-(Cyclohexyl-hydroxy-phenyl-methyO-isoxazol-S-ylmethylJ-dim ethyl^φphenyl- butyl)-ammonium bromide

The title compound, Example 30, was formed by the reaction of cyclohexyl-(5- dimethylaminomethyl-isoxazol-3-yl)-phenyl-methanol, Enantiomer 2: with (4-bromo- butyl)-benzene using an analogous method to that in example 1. LC-MS (method 10): Rt 4.59 min, m/z 447 [M]+; ¹ H NMR, 400 MHz, DMSOd ₆ : δ 0.96 - 1.33 (6H, m), 1.50 - 1.86,(8H, m), 2.17 - 2.26 (1 H, m), 2.61 (2H, t, J = 8.0 Hz), 3.03 (6H, s), 3.27 - 3.31 (2H, m), 4.78 (2H, s), 5.89 (1 H, s), 6.82 (1 H, s), 7.15 - 7.25 (4H, m), 7.26 - 7.36 (4H, m), 7.47 - 7.52 (2H, m).

Example 31

^-(Cyclohexyl-hydroxy-phenyl-methylJ-isoxazol-S-ylmethyπ -^^S^-dichloro- phenoxy)-ethyl]-dimethyl-ammonium bromide

The title compound, Example 31 , was formed by the reaction of cyclohexyl-(5- dimethylaminomethyl-isoxazol-3-yl)-phenyl-methanol, Enantiomer 2: with 4-(2-bromo- ethoxy)-1 ,2-dichIoro-benzene using an analogous method to that in example 1. LC- MS (method 10): Rt 4.51 min, m/z 503 [M]+; ¹ H NMR, 400 MHz, DMSOd ₆ : δ 0.96 - 1.32 (6H, m), 1.53 - 1.73 (4H, m), 2.18 - 2.27 (1 H, m), 3.15 (6H, s), 3.77 - 3.84 (2H, m), 4.53 - 4.60 (2H, m), 4.91 (2H, s), 5.90 (1 H, s), 6.85 (1 H, s), 7.03 (1 H, dd, J = 8.8, 2.9 Hz), 7.20 - 7.26 (1 H, m), 7.29 - 7.37 (3H, m), 7.47 - 7.52 (2H, m), 7.59 (1 H, d, J = 8.7 Hz).

Example 32

(2-Benzyloxy-ethyl)-[3-(cyclohexyl-hydroxy-phenyl-methyl) -isoxazol-5-ylmethyl]- dimethyl-ammonium bromide

The title compound, Example 31 , was formed by the reaction of cyclohexyl-(5- dimethylaminomethyl-isoxazol-3-yl)-phenyl-methanol, Enantiomer 2: with (2-bromo- ethoxymethyl)-benzene using an analogous method to that in example 1. LC-MS (method 10): Rt 4.15 min, m/z 449 [M]+; ¹ H NMR, 400 MHz, DMSOd ₆ : δ 0.95 - 1.28 (6H, m), 1.53 - 1.74 (4H, m), 2.15 - 2.26 (1 H, m), 3.10 (6H, s), 3.56 - 3.64 (2H, m), 3.89 - 3.97 (2H, m), 4.55 (2H, s), 4.87 (2H, s), 5.89 (1 H, s), 6.82 (1 H, s), 7.19 - 7.25 (1 H, m), 7.29 - 7.40 (7H, m), 7.46 - 7.51 (2H, m).

Example 33

^^-Chloro-benzyloxyJ-ethyll-fS-tcyclohexyl-hydroxy-phenyl -methylJ-isoxazol- 5-yImethyl]-dimethyl-ammonium bromide

The title compound, Example 33, was formed by the reaction of cyclohexyl-(5- dimethylaminomethyl-isoxazol-3-yl)-phenyl-methanol, Enantiomer 2: with 1 -(2-bromo- ethoxymethyl)-4-chloro-benzene using an analogous method to that in example 1. LC- MS (method 10): Rt 4.44 min, m/z 483 [M]+; ¹ H NMR, 400 MHz, DMSOd ₆ : δ 0.97 - 1.30 (6H, m), 1.54 - 1.73 (4H, m), 2.16 - 2.26 (1 H, m), 3.11 (6H, s), 3.58 - 3.64 (2H, m), 3.92 - 3.97 (2H, m), 4.54 (2H, s), 4.87 (2H, s), 5.88 (1 H, s), 6.81 (1 H, s), 7.22 (1 H, t, J = 7.2 Hz), 7.30 - 7.44 (6H, m), 7.46 - 7.52 (2H, m)

Example 34

^-(S-Chloro-phenylJ-ethyll-fS^cyclohexyl-hydroxy-phenyl-m ethylJ-isoxazol-S- ylmethylj-dimethyl-ammonium bromide

The title compound, Example 34, was formed by the reaction of cyclohexyl-(5- dimethylaminomethyl-isoxazol-3-yl)-phenyl-methanol, Enantiomer 2: with 1-(2-bromo- ethyl)-3-chloro-benzene using an analogous method to that in example 1. LC-MS (method 10): Rt 4.28 min, m/z 453 [M]+; ¹ H NMR, 400 MHz, DMSOd ₆ : δ 0.97 - 1.31 (6H, m), 1.54 - 1.73 (4H, m), 2.17 - 2.27 (1 H, m), 3.09 - 3.18 (8H, m), 3.47 - 3.54 (2H, m), 4.90 (2H, s), 5.91 (1 H, s), 6.91 (1 H, s), 7.20 - 7.45 (7H, m), 7.47 - 7.53 (2H, m).

Example 35 [3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1,2,4]oxadiazol-5-ylm ethyl]-[3-(3,4- dichloro-phenoxy)-propyl]-dimethyl-ammonium bromide

The title compound was formed by the reaction of cyclohexyl-(5-dimethylaminomethyl- [1 ,2,4]oxadiazole-3-yl)-phenyl-methanol, Enantiomer 1 (Example 8, Step c) with 4-(3- bromo-propoxy)-1 ,2-dichloro-benzene using an analogous method to that in example 1 : LC-MS (method 10): R, 4.61 min, m/z 518.2 [M]+; ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 0.94 - 1.30 (6H, m), 1.46- 1.71 (4H, m), 2.20 - 2.30 (3H, m), 3.24 (6H ₁ s), 3.53 - 3.61 (2H, m), 4.04 (2H, t, J = 5.9 Hz) ₁ 5.12 (2H, s), 6.09 (1 H, s), 6.92 - 6.96 (1 H, m), 7.20 - 7.26 (2H, m), 7.32 (2H, t, J = 7.6 Hz), 7.51 -7.58 (3H, m).

Example 36

(2-Benzyloxy-ethyl)-[3-(cyclohexyl-hydroxy-phenyl-methyl) -[1,2,4]oxadiazol-5- ylmethyl]-dimethyl-ammonium bromide

The title compound was formed by the reaction of cyclohexyl-(5-dimethylaminomethyl- [1 ,2,4]oxadiazole-3-yl)-phenyl-methanol, Enantiomer 1 (Example 8 Step c) with (2- bromo-ethoxymethyl)-benzene using an analogous method to that in example 1 : LC- MS (method 10): R _t 4.29 min, m/z 450.2 [M]+; ¹ H NMR, 400 MHz, DMSOd ₆ : δ 0.95- 1.34 (6H ₁ m), 1.46- 1.73 (4H, m), 2.21 - 2.31 (1 H, m), 3.27 (6H, s), 3.74 - 3.80 (2H, m), 3.90-3.97 (2H, m), 4.51 (2H, s), 5.15 (2H, s), 6.08 (1 H, s), 7.21 - 7.27(1 H, m), 7.27-7.39(7H, m), 7.50-7.56 (2H, m ).

Example 37

^-(Cyclohexyl-hydroxy-phenyl-methylHI^^Joxadiazol-δ-ylme thyll-p^jS- dichloro-phenoxy)-propyl]-dimethyl-ammonium bromide

The title compound was formed by the reaction of cyclohexyl-(5-dimethylaminomethyl- [1 ,2,4]oxadiazole-3-yl)-phenyl-methanol, Enantiomer 1 (Example 8 Step c) with 1 -(3- bromo-propoxy)-2,3-dichloro-benzene using an analogous method to that in example 1 : LC-MS (method 10): R, 4.46 min, m/z 518.2 [M]+; ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 0.93- 1.29 (6H, m), 1.44- 1.68 (4H, m), 2.21 - 2.37 (3H, m), 3.25 (6H, s), 3.57 - 3.64 (2H, m), 4.15 (2H, J= 6.0Hz, t), 5.15 (2H, s), 6.08 (1 H, s), 7.14 - 7.38(6H, m), 7.54 (2H, d, J = 8.3 Hz).

Example 38

^-(^Chloro-benzyloxyJ-ethylJ-tS^cycIohexyl-hydroxy-phenyl -methyl)- [1 ,2,4]oxadiazol-5-ylmethyl]-dimethyl-ammonium bromide

The title compound was formed by the reaction of cyclohexyl-(5-dimethyIaminomethyl- [1 ,2,4]oxadiazole-3-yl)-phenyl-methanol, Enantiomer 1 (Example 8 Step c) with 1-(2- bromo-ethoxymethyl)-4-chloro-benzene using an analogous method to that in example 1 : LC-MS (method 10): R _t 4.32min, m/z 484.2 [M]+; ¹ H NMR 500 MHz, DMSO-d ₆ : δ 0.96 - 1.29 (6H, m), 1.44 - 1.72 (4H, m), 2.26 (1 H, t, J = 10.3 Hz), 3.26 (6H, s), 3.74 - 3.80 (2H, m), 3.91 - 3.97 (2H, m), 4.49 (2H, s), 5.14 (2H, s), 6.08 (1 H, s), 7.24 (1 H, t, J = 6.9 Hz), 7.30 - 7.37 (4H, m), 7.41 (2H, d, J = 7.6 Hz), 7.53 (2H, d, J = 7.6 Hz).

Example 39

[S^Cyclohexyl-hydroxy-phenyl-methylJ-ti^^Joxadiazol-δ-yl methylJ-dimethyl^- phenyl-butyl)-ammonium bromide

The title compound was formed by the reaction of cyclohexyl-(5-dimethylaminomethyl- [1 ,2,4]oxadiazole-3-yl)-phenyl-methanol, Enantiomer 1 (Example 8 Step c) with (4- bromo-butyl)-benzene using an analogous method to that in example 1 : LC-MS (method 10): R _t 4.25min, m/z

448.3 [M]+; ¹ H NMR, 400 MHz, DMSO-Cl ₆ : δ 0.94 - 1.31 (6H, m), 1.45 - 1.72 (6H, m), 1.72 - 1.83 (2H, m), 2.21 - 2.31 (1 H, m), 2.56 - 2.64 (2H, m), 3.18 (6H, s), 3.37 - 3.47 (2H, m), 5.05 (2H, s), 6.09 (1 H, s), 7.17 - 7.26 (4H, m), 7.26 - 7.36 (4H, m), 7.54 (2H, d, J = 7.8 Hz).

Example 40

[S-tCyclohexyl-hydroxy-phenyl-methylJ-ti^^loxadiazol-δ-y lmethyll-dimethyl^- phenethyloxy-ethyl)-ammonium bromide

The title compound was formed by the reaction of cyclohexyl-(5-dimethylaminomethyl- [1 ,2,4]oxadiazole-3-yl)-phenyl-methanol, Enantiomer 1 (Example 8 Step c) with [2-(2- bromo-ethoxy)-ethyl]-benzene using an analogous method to that in example 1 : LC- MS (method 10): R, 4.47min, m/z 464.2 [M]+; ¹ H NMR 500 MHz, DMSOd ₆ : δ 0.97 - 1.30 (6H ₁ m), 1.45 - 1.75 (4H, m), 2.22 - 2.31 (1 H, m), 2.75 - 2.81 (2H, m), 3.16 (6H, s), 3.64 (2H, t, J = 6.5 Hz), 3.67 - 3.72 (2H, m), 3.84 - 3.89 (2H, m), 5.01 (2H, s), 6.09 (1 H ₁ s), 7.15 (1H, t, J = 6.8 Hz), 7.18 - 7.28 (5H, m), 7.34 (2H, t, J = 7.4 Hz), 7.54 (2H, d, J = 8.0 Hz).

Example 41

[3-(Cyclohexyl-hydroxy-phenyl-methyl)-[1,2,4]oxadiazol-5- ylmethyl]-[2-(3,4- dichlororbenzyloxy)-ethyl]-dimethyl-ammonium bromide

The title compound was formed by the reaction of cyclohexyl-(5-dimethylaminomethyl- [1 ,2,4]oxadiazole-3-yl)-phenyl-methanol, Enantiomer 1 (Example 8 Step c) with 4-(2- bromo-ethoxymethyl)-1 ,2-dichloro-benzene using an analogous method to that in example 1 : LC-MS (method 10): R _t 4.83min, m/z 518.2 [M]+; ¹ H NMR 400 MHz, DMSOd ₆ : δ 0.95 - 1.28 (6H ₁ m), 1.44 - 1.52 (1 H, m), 1.53 - 1.71 (3H, m), 2.20 - 2.30 (1 H, m), 3.27 (6H, s), 3.75 - 3.81 (2H, m), 3.92 - 3.99 (2H, m), 4.50 (2H, s), 5.15 (2H, s), 6.07 (1 H, s), 7.24 (1 H, t, J = 7.4 Hz), 7.28 - 7.36 (3H, m), 7.52 (2H, d, J = 7.6 Hz), 7.57 - 7.64 (2H, m).

Example 42

^-(Cyclohexyl-hydroxy-phenyl-methylJ-ti^^loxadiazol-δ-yl methyll-p-tS^- dichloro-phenoxy)-ethyl]-dimethyl-ammonium bromide

The title compound was formed by the reaction of cyclohexyl-(5-dimethylaminomethyl- [1 ,2,4]oxadiazole-3-yl)-phenyl-methanol, Enantiomer 1 (Example 8 Step c) with 4-(2- bromo-ethoxy)-1 ,2-dichloro-benzene using an analogous method to that in example 1 : LC-MS (method

10): R, 4.74min, m/z 504.1 [M]+; ¹ H NMR 400 MHz, DMSOd ₆ : δ 0.96 - 1.30 (6H, m), 1.47 - 1.71 (4H, m), 2.22 - 2.31 (1 H, m), 3.31 (6H, s), 3.94 - 4.01 (2H, m), 4.52 - 4.60 (2H, m), 5.19 (2H, s), 6.09 (1H, s), 6.92 - 6.98 (1H, m), 7.21 - 7.29 (2H, m), 7.29 - 7.37 (2H, m), 7.51 - 7.61 (3H, m).

Example 43

[2-(3-Chloro-phenyl)-ethylH3-(cyclohexyl-hydroxyφhenyl-m ethyl)- [1 ,2,4]oxadiazol-5-ylmethyl]-dimethyl-ammonium bromide

The title compound was formed by the reaction of cyclohexyl-(5-dimethylaminomethyl- [1 ,2,4]oxadiazole-3-yl)-phenyl-methanol, Enantiomer 1 (Example 8 Step c) with 1-(2- bromo-ethyl)-3-chloro-benzene using an analogous method to that in example 1 : LC- MS (method

10): R, 4.13min, m/z 454.2 [M]+; ¹ H NMR 300 MHz, DMSOd ₆ : δ 0.93 - 1.32 (6H, m), 1.43 - 1.72 (4H, m), 2.20 - 2.33 (1 H, m), 3.09 - 3.20 (2H, m), 3.28 (6H, s), 3.57 - 3.72 (2H, m), 5.16 (2H, s), 6.10 (1 H, s), 7.20 - 7.35 (4H, m), 7.35 - 7.44 (3H, m), 7.49 - 7.58 (2H, m).

Example 44

[5-(Hydroxy-diphenyl-methyl)-[1,2,4]thiadiazol-3-ylmethyl ]-dimethyl-(3-phenoxy- propyl)-ammonium bromide

a. 3-Methyl-H ,2,41thiadiazole-5-carboxylic acid ethyl ester

A solution of ethyl thiooxamate (0.73 g, 5.5 mmol) in DCM (4 ml) was treated with dimethylacetamide dimethyl acetal (0.96 ml, 6.6 mmol), stirred at room temperature for 5 mins, evaporated in vacuo, redissolved in methanol (10 ml) and treated with pyridine (0.89 ml) and a solution of hydroxylamine-O-sulfonic acid (0.74 g, 6.5 mmol) in methanol (4 ml) and stirred at room temperature for 1 h. The reaction mixture was taken up in ethyl acetate, washed with water, brine, dried (MgSO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography eluting with 10- 15% ethyl acetate-cyclohexane gave the title compound (217 mg, 23%) as a yellow semi-solid. ¹ H NMR 300 MHz δ (ppm)(CHCI ₃ -d): 1.46 (3 H, t, J = 7.14 Hz), 2.79 (3 H, s), 4.53 (2 H, q, J = 7.14 Hz).

b. (3-Methyl-π ,2,41thiadiazol-5-vD-diphenyl-methanol

A solution of the foregoing compound (500 mg, 2.9 mmol) in THF (15 ml) was cooled to -10 ⁰ C and treated dropwise with a solution of phenylmagnesium bromide (3.0 M in ether, 2.03 ml, 6.1 mmol). After 20 mins at room temperature, the reaction mixture was quenched with 0.2 N HCI and ether. The layers were separated and the organic layer was washed with sodium bicarbonate solution, brine, dried (MgSO ₄ ), filtered and evaporated in vacuo to give a white powder. Trituration with ether gave the title compound (400 mg, 49%) as a white solid. ¹ H NMR 300 MHz δ (ppm)(CHCI ₃ -d): 2.66 (3 H, s), 3.75 (1 H, s), 7.32-7.45 (10 H, m).

c. (3-Dimethylaminomethyl-π ,2,41thiadiazol-5-yl)-diphenyl-methanol

A solution of the foregoing compound (196 mg, 0.70 mmol) in 1 ,2-dichloroethane (2 ml) was treated with NBS (150 mg, 0.84 mmol) and AIBN (1 1 mg, 0.07 mmol) and heated at 90 ⁰ C for 2h. A further quantity of NBS (25 mg) and AIBN (1 mg) was added and the reaction was heated for a further 2h. The reaction mixture was cooled to room temperature and evaporated in vacuo. The resulting crude solid was taken up in THF and treated with dimethylamine (2.0 M in THF, 1 ml). The reaction mixture was evaporated in vacuo. Purification by SCX and flash column chromatography (eluting with cyclohexane-ethyl acetate (1 :1 )) gave the title compound (35 mg, 15%) as a solid. ¹ H NMR 300 MHz δ (ppm)(CHCI ₃ -d): 2.22-2.34 (6 H, m), 3.71 -3.80 (2 H, m), 4.40 (1 H, brs), 725-7 Al (10 H, m).

d. | ^" 5-(Hvdroxy-diphenyl-methyl)-π ,2,41thiadiazol-3-ylmethyll-dimethyl-(3-phenoχy- propyO-ammonium bromide

A solution of the foregoing compound (40 mg, 0.12 mmol) and 3-phenoxypropyl bromide (0.2 ml, 1.23 mmol) in acetonitrile/chloroform (1 :1 ; 1 ml) was heated at 50 ⁰ C for 72 h. The reaction mixture was evaporated in vacuo and purified by flash column chromatography eluting with 2-12% MeOH-DCM to give the title compound (50 mg, 75%) as a white solid. LC-MS (Method 4): R, 8.21 min, m/z 460 [M-Br]+. ¹ H NMR 400 MHz δ (ppm)(CHCI ₃ -d): 2.23-2.32 (2 H, m), 3.46 (6 H, s), 3.65-3.76 (2 H, m), 3.96 (2 H, t, J = 5.50 Hz), 5.01 (2 H, s), 5.17 (1 H, s), 6.72-6.81 (2 H, m), 6.93 (1 H, t, J = 7.33 Hz), 7.19-7.34 (8 H, m), 7.40-7.43 (4 H, m).

Example 45

[δ^Cyclohexyl-hydroxy-phenyl-methylJ-fi^^lthiadiazol-S-y lmethyll-dimethyl^S- phenoxy-propyl)-ammonium formate

a. 3-Methyl-f1 ,2,41thiadiazole-5-carbaldehvde

A solution of the compound obtained in Example 1 , Step a (2.0 g, 11.6 mmol) in DCM (100 ml) was cooled to -78 ⁰ C. A solution of DIBAL-H (1.0 M in toluene, 20 ml, 20 mmol) was added dropwise. The reaction was stirred for an additional 1 h, and then quenched at -78 ⁰ C by the dropwise addition of methanol/acetic acid (10 ml/5 ml), keeping the temperature below -70 ⁰ C. The reaction mixture was warmed to room temperature and partitioned between 1 N HCI and DCM. The organic phase was washed with water, dried (MgSO ₄ ), filtered and evaporated to give the title compound as a component of a mixture of product/starting material/toluene in the approximate molar ratio (1 :1 :2). Yield of title compound approximately 20-30%.

b. (3-Methyl-H ,2,41thiadiazol-5-yl)-phenyl-methanone A solution of the foregoing compound (approximately 0.5 g) in THF (10 ml) was cooled to 0 ⁰ C, and treated with an excess of phenylmagnesium bromide solution (3.0 M in ether). The reaction mixture was warmed to room temperature and quenched by the addition of NH ₄ CI solution and extracted with ethyl acetate. The organic phase was dried (MgSO ₄ ), filtered and evaporated in vacuo to give the title compound as a crude product (180 mg). This was dissolved in DCM (5 ml) and treated with pyridinium chlorochromate (372 mg, 1.7 mmol) and stirred at room temperature overnight. The reaction mixture was washed with water and the organic phase separated, dried (MgSO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography eluting with 4:1 cyclohexane/ethyl acetate gave the title compound (180 mg) as a white solid. ¹ H NMR 300 MHz δ (ppm)(CHCI ₃ -d): 2.83 (3 H, s), 7.52-7.59 (2 H, m), 7.65-7.72 (1 H, m), 8.51 (2 H, dd, J = 8.24, 1.49 Hz).

c. Cvclohexyl-(3-methyl-H ,2,41thiadiazol-5-yl)-phenyl-methanol

A solution of the foregoing compound (80 mg, 0.39 mmol) in THF (5 ml) was cooled to -10 °C, treated with a solution of cyclohexylmagnesium chloride (2.0 M in ether, 0.6 ml, 1.2 mmol), warmed to room temperature and stirred for 10 min. The reaction was quenched with NH ₄ CI and ether. The organic layer was separated and washed with brine, dried (MgSO ₄ ), filtered and evaporated in vacuo. Purification by column chromatography gave the title compound (80 mg, 72%) as a colorless gum. ¹ H NMR 300 MHz δ (ppm)(CHCI ₃ -d): 1.40-1.10 (8 H, m), 1.69-1.62 (3 H, m), 2.49 (1 H, td, J = 11.54, 3.40 Hz), 2.64 (3 H, s), 3.14 (1 H, s), 7.31-7.22 (1 H, m), 7.40-7.31 (2 H, m), 7.74-7.67 (1 H, m).

d. Cvclohexyl-O-dimethylaminomethyl-fi ^λlthiadiazol-δ-vD-phenyl-methanol A solution of the foregoing compound (80 mg, 0.28 mmol), AIBN (4.5 mg, 0.028 mmol) and NBS (60 mg, 0.34 mmol) in 1 ,2-dichloroethane (1 ml) was heated at 90 ⁰ C overnight. The reaction mixture was cooled, evaporated in vacuo and taken up in a solution of dimethylamine in THF (2.0 M, 2 ml), stirred for 1 h and evaporated in vacuo. Purification by SCX and flash column chromatography gave the title compound (15 mg, 13%) in a purity of about 80%. ¹ H NMR 300 MHz δ (ppm)(CHCI ₃ - d): 1.72-0.90 (10 H, m), 2.42 (6 H, s), 2.56-2.48 (1 H, m), 3.30 (1 H, brs), 3.84 (2 H, s), 7.39-7.22 (3 H, m), 7.75-7.66 (2 H, m).

e. fS-fCvclohexyl-hvdroxy-phenyl-methvD-π ^^ithiadiazol-S-ylmethyll-dimethyl-O- phenoxy-propyP-ammonium formate

A mixture of the foregoing compound (15 mg), phenoxypropyl bromide (0.2 ml) and MeCN-CHCI ₃ (1 :1 , 1 ml) was heated at 55 ⁰ C for 24 h. The reaction mixture was evaporated in vacuo and purified by column chromatography (2-12% MeOH-DCM). Further purification by reverse-phase HPLC gave the title compound (5 mg, 20%) as a white solid. LC-MS (Method 8): R, 8.36 min, m/z 466 [M-formate]+. ¹ H NMR 400 MHz δ (ppm)(CHCI ₃ -d): 0.95-1.40 (6 H, m), 1.51 -1.68 (3 H, m), 2.32-2.36 (4 H, m), 3.34 (6 H, s), 3.66 (2 H, d, J = 9.93 Hz), 3.88-4.05 (2 H, m), 4.76-4.88 (2 H, m), 6.78 (2 H, d, J = 8.10 Hz), 6.90-6.97 (1 H, m), 7.15-7.30 (5 H, m), 7.66-7.72 (2 H, m), 8.74 (1 H, s).

Example 46 [S^Cyclohexyl-hydroxy-phenyl-methylJ-ti^.^thiadiazol-S-ylmet hyπ-dimethyl^S-

phenoxy-propyl)-ammonium bromide

a. (5-Chloro-π ,2,41thiadiazol-3-vπ-phenyl-methanone

A solution of 5-chloro-3-[phenyl-(tetrahydro-pyran-2-yIoxy)-methyl]-[1 ,2,4]thiadiazole (A. M. Macleod et al, J. Med. Chem., 1990, 33, 2052) (0.50 g, 1.62 mmol) in ethyl acetate (5 ml) and 5 N HCI (5 ml) was stirred vigorously for 1 h. The reaction mixture was diluted further with ethyl acetate and washed with sodium bicarbonate solution, brine, dried (MgSO ₄ ), filtered and evaporated in vacuo to give a crude yellow oil. This was re-dissolved in DCM (10 ml), treated with pyridinium chlorochromate (0.52 g, 2.4 mmol) and stirred at room temperature for 4 h. The reaction mixture was decanted directly onto a short silica column and eluted with DCM to give the title compound (300 mg, 83%) as a colourless oil. . ¹ H NMR 300 MHz δ (ppm)(CHCI ₃ -d): 8.25-8.14 (2 H, m), 7.72-7.63 (1 H, m), 7.58-7.50 (2 H, m).

b. Phenyl-(5-vinvK1 ,2,41thiadiazol-3-yl)-methanone A solution of the foregoing compound (300 mg, 1.23 mmol), tributyl(vinyl)tin (0.37 ml, 1.85 mmol) and Pd(PPh ₃ ) ₄ (77 mg, 0.06 mmol) in toluene (3 ml) was heated at reflux for 1 h. The reaction mixture was cooled, evaporated in vacuo and purified by flash column chromatography eluting with 5% ethyl acetate/cyclohexane to give the title compound (230 mg, 81%) as a brown oil. ¹ H NMR 300 MHz δ (ppm)(CHCI ₃ -d): 5.94 (1 H, d, J = 10.94 Hz), 6.40 (1 H, d, J = 17.59 Hz), 7.11 (1 H, dd, J = 17.62, 10.93 Hz), 7.49-7.56 (2 H, m), 7.61 -7.68 (1 H, m), 8.22-8.27 (2 H, m).

c. CvclohθXyl-phenyl-(5-vinyl-ri ,2,41thiadiazol-3-yl)-methanol

A solution of the foregoing compound (230 mg, 1.06 mmol) in THF (10 ml) was cooled to -78 ⁰ C and treated with a solution of cyclohexylmagnesium chloride (2.0 M in ether, 0.6 ml, 1.2 mmol). The reaction mixture was warmed to -30 ⁰ C, and then quenched with NH ₄ CI solution. The reaction mixture was extracted with ethyl acetate and the organic phase was dried (MgSO ₄ ), filtered, and evaporated in vacuo. Purification by column chromatography eluting with 1 :20 ethyl acetate/cyclohexane gave the title compound (100 mg, 31 %) as a brown oil. ¹ H NMR 300 MHz δ (ppm)(CHCI ₃ -d): 1.12- 1.77 (10 H, m), 2.51-2.55 (1 H, m), 4.17 (1 H, d, J = 0.72 Hz), 5.80 (1 H, d, J = 10.94 Hz), 6.24 (1 H, d, J = 17.54 Hz), 6.95 (1 H, dd, J = 17.56, 10.90 Hz), 7.21-7.25 (1 H, m), 7.28-7.35 (2 H, m), 7.74-7.79 (2 H, m).

d. CyclohexyHδ-dimethylaminomethyl-ri ^λlthiadiazol-S-vO-phenyl-methanol A solution of the foregoing compound was dissolved in DCM (5 ml) and methanol (1 ml), cooled to -78 ⁰ C and treated with an excess of ozone. The reaction mixture was quenched with an excess of dimethylsulfide and allowed to warm to room temperature and stirred for 2 h. The reaction mixture was washed with water, dried (MgSO ₄ ), filtered and evaporated in vacuo. Purification by flash column chromatography gave the crude aldehyde (60 mg) as a yellow gum. A solution of the aldehyde, dimethylamine solution (2.0 M in THF, 0.15 ml, 0.3 mmol) and acetic acid (0.01 ml) in 1 ,2-dichloroethane (1 ml) was treated with sodium triacetoxyborohydride (46 mg, 0.22 mmol) and stirred at room temperature overnight. The reaction mixture was diluted with water and DCM, passed through a phase-separation cartridge and evaporated in vacuo. Purification by SCX gave the title compound (40 mg, 36%, 2 steps) as a pale yellow oil. ¹ H NMR 300 MHz δ (ppm)(CHCI ₃ -d): 1.11 -1.64 (10 H, m), 2.40 (6 H, s), 2.45-2.56 (1 H, m), 3.85 (2 H, s), 4.21 (1 H, s), 7.17-7.34 (3 H, m), 7.73-7.78 (2 H, m).

e. fS-fCvclohexyl-hvdroxy-phenyl-methvD-π ^λlthiadiazol-δ-ylmethvn-dimethyl-O- phenoxy-propyD-ammonium bromide

A solution of the foregoing compound (40 mg, 0.12 mmol), and 3-phenoxypropyl bromide (0.2 ml) in MeCN-CHCI ₃ (1 :1 , 1 mi) was heated at 50 ⁰ C for 100 h. The reaction mixture was evaporated in vacuo and purified by column chromatography (2- 12% MeOH-DCM) to give the title compound (18 mg, 27%) as an off-white foam. LC- MS (Method 4): R ₁ 8.73 min, m/z 466 [M-Br]+. ¹ H NMR 400 MHz δ (ppm)(CHCI ₃ -d): 1.07-1.72 (10 H, m), 2.33-2.53 (3 H, m), 3.44 (3 H, s), 3.48 (3 H, s), 3.77-3.92 (2 H, m), 3.97 (1 H, s), 4.01 -4.08 (2 H, m), 5.79 (1 H, d, J = 14.30 Hz), 5.99 (1 H, d, J = 14.30 Hz), 6.80-6.83 (2 H, m), 6.89-6.96 (1 H, m), 7.11-7.16 (1 H, m), 7.19-7.27 (4 H, m), 7.62-7.67 (2 H, m).

Example 47

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1,3,4]thiadiazol-2 -ylmethyl]-dimethyl-(3- phenoxy-propyl)-ammonium bromide

a (Cvclohexyl-fδ-dimethylaminomethyl-fi .SλIthiadiazol^-vD-phenyl-methanol A solution of cyclohexyl-hydroxy-phenyl-acetic acid N'-(2-dimethylaminoacetyl)- hydrazide (prepared as described in Example 55 (1 g, 3 mmol) and imidazole (1.144 g, 16.8 mmol) in dry dimethylformamide (25 ml) under nitrogen was stirred at room temperature and treated dropwise with chlorotrimethylsilane (1.5 ml, 12 mmol). After stirring for 72 hours at room temperature, further imidazole (0.3813 g, 5.6 mmol) was added followed by dropwise addition of chlorotrimethylsilane (0.5 ml, 4 mmol). After stirring for a further 18 hours the reaction mixture was poured onto ice/water and

extracted three times with diethyl ether. The combined organic extracts were washed with water, brine, dried (MgSO ₄ ), filtered and evaporated in vacuo. The resulting crude material was treated with dry toluene (90 ml) and Lawesson's reagent (2.4 g, 5.93 mmol) and stirred and refluxed under a nitrogen atmosphere for 90 minutes. The crude reaction mixture was purified by SCX-2 cartridge followed by chromatography using a silica isolute cartridge eluting with 0-50% ethyl acetate in DCM and then preparative HPLC using a Phenomenex column and 0-70% acetonitrile in water containing 0.1% formic acid; flow rate 10 ml/min. The title compound was obtained by treatment of the resulting formate salt with DCM and saturated sodium bicarbonate solution. The aqueous layer was extracted four more times with DCM. The combined organic extracts were dried (MgSO ₄ ), filtered and evaporated in vacuo to give the title compound (141.7 mg, 14.4%) as a white foam.

¹ H NMR, 400 MHz, CDCI ₃ : δ 1.07-1.5 (7 H, m), 1.61 -1.8 (3 H, m), 2.32 (6 H, s), 2.42- 2.52 (1 H, m), 3.56 (1 H, s), 3.80 (2 H, s), 7.22-7.26 (1 H, m), 7.32-7.36 (2 H, m), 7.64- 7.69 (2 H, m).

The enantiomers were separated using a 250 x 20 mm Chiralpak ® IA column packed with amylase tris(3,5-dimethylphenylcarbamate) immobilized on 5 μm silica gel. The column was eluted with 10% IPA in heptane buffered with 0.1% diethylamine. Flow rate, 18 ml/min. Wavelength, 220nm. Enantiomer 1 : R _t 9.24 mins, enantiomer 2 : R _t 12.5 mins.

Enantiomer 1 ¹ H NMR, 400 MHz, CDCI ₃ : δ 1.07-1.42 (6H, m), 1.46-1.55 (1 H, m), 1.63- 1.81 (3H, m), 2.32 (6H, s), 2.43-2.52 (1 H, m), 3.56 (1 H, s), 3.80 (2H,s), 7.21-7.26 (1 H, m), 7.31-7.38 (2H, m), 7.64-7.69 (2H, m). Enantiomer 2 ¹ H NMR, 400 MHz, CDCI ₃ : δ 1.06-1.41 (6H, m), 1.45-1.53 (1 H, m), 1.62- 1.80 (3H, m), 2.31 (6H, s), 2.42-2.52 (1 H, m), 3.56 (1 H, s), 3.80 (2H,s), 7.21 -7.26 (1 H, m), 7.31-7.38 (2H, m), 7.64-7.69 (2H, m).

b.fδ-fCvclohθxyl-hvdroxy-phθnyl-methvD-fi .SλIthiadiazol^-ylmethyli-dimethyl-O- phenoxy-propyD-ammonium bromide

A solution of (cyclohexyl-Cδ-dimethylaminomethyl-fi ,3,4]thiadiazol-2-yl)-phenyl- methanol enantiomer 1 (31.7 mg, 0.096 mmol) and 3-phenoxypropyl bromide (0.08 mi, 0.5 mmol) in acetonitrile (0.45 ml) and chloroform (0.65 ml) was heated at 50 ⁰ C for 90 hours. A precipitate formed which was collected, washed with diethyl ether and dried in vacuo to give the title compound as a white solid (15.5 mg, 29.5%)

LC-MS (Method 4): R ₁ 8.62 min, m/z 466 [M] ⁺ .

¹ H NMR δ (ppm)(CH ₃ OH-dG): 1.08-1.46 (7 H, m), 1.62-1.77 (3 H, m), 2.33-2.42 (2 H, m), 2.63 (1 H, t, J = 11.47 Hz), 3.25 (6 H, s), 3.62-3.68 (2 H, m), 4.07-4.13 (2 H, m),

5.09 (2 H, s), 6.87-6.97 (3 H, m), 7.21-7.29 (3 H, m), 7.30-7.35 (2 H, m), 7.68-7.72 (2

H, m), 7.89 (1 H, s).

Example 48 [5-Cyclohexyl-hydroxy-phenyl-methyl)-[1,2,4]oxadiazol-3-ylme thyl]-[2-(3,4- dichloro-phenoxy)-ethyl]-dimethyl-ammonium formate

NC ^~ λ

N-boc

a. Cyanomethyl-methyl-carbamic acid tert-butyl ester

To a suspension of methylaminoacetonitrile hydrochloride (11.45 g, 107.4 mmol) in DCM (100 ml_) was added triethylamine (31.5 mL, 226 mmol), followed by di-tert-butyl dicarbonate (22.3 g, 102.2 mmol) and the reaction mixture was stirred at room temperature over the weekend. The reaction mixture was filtered, concentrated in vacuo and purified over silica using a gradient of 10-20% ethyl acetate/cyclohexane. The pure fractions were combined and concentrated to afford the title compound (11.94 g, 69%) as a colourless liquid. LC-MS (Method 5): 2.82 min, [M+H] ⁺ 171.1.

b. (/V-HvdroxycarbamimidoylmethvD-methyl-carbamic acid fert-butyl ester

A solution of cyanomethyl-methyl-carbamic acid tert-butyl ester (6.01 g, 35.3 mmol) and hydroxylamine (5.13 g, 77.6 mmol, 50% solution in water) in ethanol (30 mL) was

heated at reflux for 1 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was azeotroped twice with ethanol, then once with isopropyl alcohol to afford a white solid. This was triturated with ether and collected by filtration, then dried under vacuum at room temperature to afford the title compound (5.27 g) as a white solid. A further crop could be obtained from the mother liquors by the same work-up procedure. Total yield 6.93 g (96%). ¹ H NMR (CDCI ₃ ): 1.48 (S, 9 H), 2.87 (s, 3 H), 3.84 (s, 2 H), 4.73 (brs., 1 H), 5.09 (bra., 2 H).

c. fδ-^vclohexyl-hvdroxy-phenyl-methvπ-π ^λloxadiazol-S-ylmethvn-methyl-carbamic acid te/t-butyl ester

1 ,1'-Carbonyldiimidazole (2.08 g, 12.8 mmol) was added to a solution of cyclohexylmandelic acid (2.50 g, 10.7 mmol) in 1 ,4-dioxane (100 mL) and heated at 80 ⁰ C for 1 h. After allowing to cool to room temperature, (N- hydroxycarbamimidoylmethyl)-methyl-carbamic acid tert-butyl ester (2.08 g, 12.8 mmol) was added and the mixture was heated at reflux under nitrogen atmosphere for 3.5 days. The reaction mixture was concentrated in vacuo to afford a dark brown oil, which was purified over silica using a gradient of 0-25% acetone/cyclohexane. The pure fractions were combined and concentrated in vacuo to afford the product (825 mg, 19%) as a light yellow viscous oil. LC-MS (Method 5): 4.37 min, [M-H] ^" 400.3.

d. Cyclohexyl-O-methylaminomethyl-ri ^^loxadiazol-δ-vD-phenyl-methanol

To a solution of [5-(cyclohexyl-hydroxy-phenyl-methyl)-[1 ,2,4]oxadiazol-3-ylmethyl]- methyl-carbamic acid tert-butyl ester (795.5 mg, 1.98 mmol) in DCM (10 ml) was added TFA (3 ml). After stirring at room temperature for 5 h the solvent was removed in vacuo and the residue partitioned between DCM (10 ml) and a saturated sodium

hydrogen carbonate solution (10 ml). The organic layer was separated, dried (MgSO ₄ ) and the solvent removed in vacuo. Purification by column chromatography using DCM/methanol (9:1) as eluent afforded the title compound as a yellow oil (564.4 mg, 95%). LC-MS (Method 5): R ₁ 2.24 min, m/z 303 [MH] ⁺ .

e. Cvclohexyl-O-dimethylaminomethyl-fi ^λioxadiazol-S-vD-phenyl-methanol

To a DCE solution (15 ml) of cyclohexyl-(3-methylaminomethyl-[1 ,2,4]oxadiazol-5-yl)- phenyl-methanol (666.4 mg, 2.21 mmol) at 0 ⁰ C was added formaldehyde (37% in water, 359 μl, 4.42 mmol). After stirring for 10 min, sodium triacetoxyborohydride (516 mg, 2.44 mmol) was added portionwise. The reaction was next allowed to warm to room temperature and stirred overnight at room temperature. Next the reaction was partitioned by the addition of a saturated sodium hydrogen carbonate solution (10 ml). The organic layer was separated, dried (MgSO ₄ ) and the solvent removed in vacuo. The residue was purified by column chromatography using DCM/methanol (9:1 ) as eluent to afford the title compound as a yellow oil (482 mg, 69%). The enantiomers were separated via chiral resolution method 7 using 6% IPA in heptane as the eluent. Enantiomer 1 : R _t 6.85 min, enantiomer 2 : R, 6.90 min.

f. rδ-Cvclohexyl-hydroxy-phenyl-methvD-fi ^λioxadiazol-S-ylmethvn-^-fSλ-dichloro- phenoxy)-ethyll-dimethyl-ammonium formate

The title compound was prepared from cyclohexyl-(3-dimethylaminomethyl-

[1 ,2,4]oxadiazol-5-yl)-phenyl-methanol (enantiomer 1 ) in a similar manner to that described in Example 1 and purified by HPLC. LC-MS (Method 4): R _t 9.15 min, m/z 504 [Mf. ¹ H NMR (400 MHz) δ (ppm)(CHCI ₃ -d): 0.95-1.37 (7 H, m), 1.53-1.73 (3 H,

m), 2.24 (1 H, t, J = 10.25 Hz) ₁ 3.35 (6 H ₁ s), 4.10 (2 H, s), 4.45 (2 H, s), 4.86-5.06 (2 H, m), 6.77 (1 H, del, J = 8.89, 2.83 Hz), 7.02 (1 H, d, J = 2.80 Hz), 7.12-7.30 (4 H, m), 7.49 (2 H, d, J = 7.63 Hz) ₁ 8.45 (1 H, s).

Example 49

[2-(4-Chloro-benzyloxy)-ethyl]-[5-(cyclohexyl-hydroxy-phe nyl-methyl)- [1 ,2,4]oxadiazol-3-ylmethyl]-dimethy!-ammonium formate

The title compound was formed by the reaction of cyclohexyl-(3-dimethylaminomethyl- [1 ,2,4]oxadiazol-5-yl)-phenyl-methanol (enantiomer 1 ) with 1 -(2-bromoethoxymethyl)- 4-chloro-benzene using an analogous method to that in Example 1 and purified by HPLC. LC-MS (Method 4): R _t 8.90 min, m/z 484 [M]+. ¹ H NMR (400 MHz) δ (ppm)(CHCI ₃ -d): 0.98-1.37 (7 H, m), 1.53-1.77 (3 H, m), 2.26 (1 H, t, J = 10.17 Hz), 3.29 (6 H, s), 3.90 (4 H, d, J = 13.47 Hz), 4.47 (2 H, s), 4.82-4.93 (1 H, m), 4.95 (1 H, d, J = 14.08 Hz), 7.16-7.32 (7 H, m), 7.51 (2 H, d, J = 7.64 Hz), 8.52 (1 H, s).

Example 50 [5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1,2,4]oxadiazol-3-ylm ethyl]-[2-(3,4- dichloro-benzyloxy)-ethyl]-dimethyl-ammonium formate

The title compound was formed by the reaction of cyclohexyl-(3-dimethylaminomethyl- [1 ,2,4]oxadiazol-5-yl)-phenyl-methanol (enantiomer 1 ) with 4-(2-bromoethoxymethyl)- 1 ,2-dichlorobenzene using an analogous method to that in Example 1 and purified by HPLC. LC-MS (Method 4): R _t 9.19 min, m/z 518 [M] ⁺ . ¹ H NMR (400 MHz) δ

(ppm) (CHCI ₃ -Cl): 0.99-1.39 (6 H, m), 1.55-1.74 (4 H, m), 2.27 (1 H ₁ 1, J = 10.27 Hz), 3.31 (6 H, s), 3.91 (2 H, s), 3.95 (2 H ₁ s), 4.47 (2 H ₁ s), 4.83-5.03 (2 H ₁ m), 7.10-7.27 (4 H, m), 7.35-7.42 (2 H ₁ m), 7.51 (2 H ₁ d, J = 7.63 Hz) ₁ 8.52 (1 H, s).

Example 51

[S^CycIohexyl-hydroxy-phenyl-methylJ-fi^^joxadiazol-S-ylm ethyll-dimethyl^- phenethyloxy-ethyl)ammonium formate

The title compound was formed by the reaction of cyclohexyl-(3-dimethylaminomethyl- [1 ,2,4]oxadiazol-5-yl)-phenyl-methanol (enantiomer 1 ) with [2-(2-bromo-ethoxy)-ethyI]- benzene using an analogous method to that in Example 1 and purified by HPLC. LC- MS (Method 4): R _t 8.59 min, m/z 464 [M] ⁺ . ¹ H NMR (400 MHz) δ (ppm)(CHCI ₃ -d): 1.02- 1.42 (8 H ₁ m), 1.56-1.77 (4 H ₁ m), 2.23-2.34 (1 H, m), 3.14 (6 H ₁ s), 3.70 (2 H, t, J = 6.51 Hz) ₁ 3.84 (5 H, s), 4.65-4.87 (2 H, m), 7.12-7.18 (3 H, m), 7.18-7.32 (5 H, m), 7.52-7.57 (2 H, m), 8.57 (1 H ₁ s).

Example 52

[5-(Hydroxy-diphenyl-methyl)-isoxazol-3-ylmethyl]-dimethy l-(3-phenoxy-propyl)- ammonium bromide

a. 3-(Tetrahvdro-pyran-2-yloxymethyl)-isoxazole-5-carboxylic acid ethyl ester Di-tert-butyl dicarbonate (14.13 g, 64.7 mmol) and ethyl propiolate (21.17 g, 215.8 mmol) were dissolved in acetonitrile (216 ml) and DMAP (527 mg, 4.32 mmol) was added. A solution of 2-(2-nitroethoxy)tetrahydropyran (7.65 g, 43.17 mmol) in acetonitrile (216 ml) was added dropwise over a period of 1 h at room temperature

and the reaction mixture was stirred overnight. The solvent was evaporated in vacuo. Purification using CombiFlash Companion ® system over silica eluting with 0-20 % ethyl acetate in cyclohexane gave the title compound (3.0 g, 27 %) as a yellow oil. ¹ H NMR, 400 MHz, CDCI ₃ : δ 7.01 (1 H, s), 4.85 (1 H, d, J = 13 Hz), 4.75-4.70 (1 H, m), 4.67 (1 H, d, J = 13 Hz), 4.44 (2 H, q, J = 7 Hz), 3.82-3.91 (1 H, m), 3.52-3.61 (1 H, m), 1.47 - 1.90 (6 H, m), 1.42 (3 H, t, J = 7 Hz).

b. Diphenyl-r3-(tetrahvdro-pyran-2-yloxymethyl)-isoxazol-5-yll- methanol

Phenylmaganesium bromide (1.0 M in THF, 2.52 ml, 2.52 mmol) was added dropwise to 3-(tetrahydro-pyran-2-yloxymethyl)-isoxazole-5-carboxylic acid ethyl ester (0.30 g, 2.52 mmol) in THF (4 ml) at 0 ⁰ C. The reaction was stirred at room temperature for 2 h and then refluxed overnight. The reaction was cooled to room temperature and quenched with 1 N HCI. The aqueous layer was extracted with ethyl acetate and washed with sat. NaHCO ₃ , brine, dried (MgSO ₄ ), filtered and concentrated in vacuo. Purification using CombiFlash Companion® system over silica using 0-20 % ethyl acetate/cyclohexane as eluent gave the title compound (0.12 g, 27%) as an off-white solid. LCMS (Method 9): R, 3.82 min, m/z 364 [M-H] ⁺ .

c. (3-Hvdroxymethyl-isoxazol-5-yl)-diphenyl-methanol

Pyridinium tosylate (0.0074 g, 0.029 mmol) was added to a solution of diphenyl-[3- (tetrahydro-pyran-2-yloxymethyl)-isoxazol-5-yl]-methanol (0.11 g, 0.29 mmol) in IMS (3 ml). The reaction mixture was heated to 55 ⁰ C for 4.5 h. The reaction was cooled to room temperature and concentrated in-vacuo. Purification using CombiFlash Companion ® system over silica using 0-30 % ethyl acetate/cyclohexane as eluent gave the title compound (0.062 g, 76 %) as a white solid. LCMS (Method 9): R, 2.91

min, m/z 282 [M+H] ⁺ .

d. (3-Bromomethyl-isoxazol-5-yl)-diphenyl-methanol

Carbon tetrabromide (0.76 g, 2.30 mmol) and triphenylphosphine (0.60 g, 2.30 mmol) were added to a solution of (3-hydroxymethyl-isoxazol-5-yl)-diphenyI-methanol (0.43 g, 1.53 mmol) in methylene chloride (13 mL). The reaction mixture was stirred at room temperature overnight and then concentrated in vacuo. Purification using CombiFlash Companion ® system over silica using 0-40 % ethyl acetate/cyclohexane as eluent gave the title compound (501.8 mg, 95%) as a white solid. LCMS (Method 9): R _t 3.75 min, m/z 346 [M+H] ⁺ .

e. (3-Dimethylaminoethyl-isoxazol-5-yl)-diphenyl-methanol

A solution of (3-bromomethyl-isoxazol-5-yl)-diphenyl-methanol (0.12 g, 0.35 mmol) in THF (2 ml) was added dropwise to a solution of dimethylamine (2.0 M in THF, 0.52 ml, 1.05 mmol). The reaction mixture was stirred at room temperature for 1 h and then concentrated in vacuo. The residue was partitioned between ethyl acetate and water and the organic layer separated. The aqueous layer was extracted with ethyl acetate. The organic layers were combined and washed with brine, dried (MgSO ₄ ), filtered and concentrated in vacuo. Purification using CombiFlash Companion ® system over silica using 0-10% methanol/methylene chloride as eluent gave the title compound (0.042 g, 39%) as a white solid. ¹ H NMR, 400 MHz, CDCI ₃ : δ 7.37-7.29 (10 H, m), 6.03 (1 H, s), 3.50 (2 H, s), 3.46-3.27 (1 H, brs), 2.27 (6 H, s).

f. f5-(Hvdroxy-diphenyl-methyl)-isoxazol-3-ylmethyl1-dimethyl-( 3-phenoxy-propyl)- ammonium bromide

To a solution of (3-dimethylaminoethyl-isoxazol-5-yl)-diphenyl-methanol (0.034 g, 0.10 mmol) in chloroform (0.6 ml) and CH ₃ CN (0.4 ml) was added 3-phenoxypropyl bromide (0.10 ml, 0.54 mmol). The mixture was heated at 50 ⁰ C for 48 h. After this period the solvents were evaporated and the residue purified using CombiFlash Companion ® system over silica using 0-10% methanol/methylene chloride as eluent to give the title compound (0.041 g, 72%) as a white solid. LC-MS (Method 4): R, 8.03 min, m/z 443 [M-Br] ⁺ . ¹ H NMR, 400 MHz, DMSO-d ₆ : δ 7.35-7.21 (12 H, m), 7.16 (1 H, s), 6.94-6.84 (3 H, m), 6.52 (1 H, s), 4.71 (2 H, s), 4.00 (2 H, t, J = 6 Hz), 3.54-3.47 (2 H, m), 3.10 (6 H, s), 2.30-2.17 (2 H, m).

Example 53 [5-(9-Hydroxy-9H-xanthen-9-yl)-[1,2,4]oxadiazol-3-ylmethyl]- dimethyl-(3- phenoxy-propyl)-ammonium bromide

a. 2-Chloro-N-hydroxy-acetamidine Sodium ethoxide solution (21 % w/w in ethanol, 60 ml, 0.16 mol) was added dropwise to a suspension of hydroxylamine hydrochloride (11.4 g, 0.16 mol) in IMS (200 ml) at 0 ⁰ C. Stirring was continued for 10 min, then chloroacetonitrile (10 ml, 0.158 mol) was added dropwise. The reaction mixture was warmed to room temperature over 1 h, and then filtered. The filtrate was concentrated in vacuo to give the title compound (16.5 g, 96%) as a yellow solid.

b. 9H-Xanthene-9-carboxylic acid (2-chloro-1-hvdroxyimino-ethyl)-amide

A mixture of 9H-xanthene-9-carboxylic acid (1.46g, 6.5mmol), 2-chloro-N-hydroxy- acetamidine (1 g, 9.3 mmol), O-(7-azobenzotriazol-1 -yl)-1 ,1 ,3,3-tetramethyluronium hexafluorophosphate (3.0 g, 7.5 mmol) and diisopropylethylamine (3 ml) in DMF (15 ml) was stirred at room temperature for 18 h. The reaction mixture was partitioned between water and ethyl acetate. The aqueous phase was extracted with ethyl acetate. The combined organic phases were dried (Na ₂ SO ₄ ) and concentrated in vacuo to leave a yellow oil. Purification by flash column chromatography on a silica isolute cartridge eluting with pentane, 4:1 -1 :1 pentane-ether, and ethyl acetate gave the title compound as a pale orange solid, (1.5 g, 78%).

c. 3-Chloromethyl-5-(9H-xanthen-9-ylH1.2.41oxadiazole

A solution of 9H-xanthene-9-carboxylic acid (2-chloro-1 -hydroxyimino-ethyl)-amide (1.5 g, 4.75 mmol) in acetic acid was heated at reflux for 20 h. The reaction mixture was partitioned between water and diethyl ether. The aqueous layer was extracted with diethyl ether. The combined organic phases were dried (Na ₂ SO ₄ ) and concentrated in vacuo to leave an orange solid, which was taken up in DCM and filtered to remove the insoluble residues. The resulting solution was evaporated and purified by flash column chromatography on a silica isolute cartridge eluting with pentane, and 4:1-1 :1 pentane-ether.to give the title compound as an off- white solid, (350 mg, 26%). LC-MS (Method 9): Rt 3.94 min, m/z 297 [M-H] ⁺

d. 9-(3-Dimethylaminomethyl-ri ,2,41oxadiazol-5-yl)-9H-xanthen-9-ol

A solution of 3-chloromethyl-5-(9H-xanthen-9-yl)-[1 ,2,4]oxadiazole (340 mg, 1.14 mmol) in THF (10 ml) was added dropwise to a stirred solution of dimethylamine in

THF (2 M, 3 ml, 6 mmol). Stirring was continued at room temperature for 20 h. The reaction mixture was concentrated in vacuo to leave a yellow oil. Flash column chromatography eluting with 0-5% methanol-DCM gave the title compound (94 mg,

26%) as a white solid

¹ H NMR 400 MHz δ (ppm)(CHCI ₃ -d): 2.15-2.32 (6 H, m), 3.57 (2 H, s), 4.78 (1 H, s),

7.05-7.27 (4 H, m), 7.29-7.43 (2 H, m), 7.53 (2 H, dd, J = 7.89, 1.65 Hz).

e. r5-(9-Hvdroxy-9H-xanthen-9-vh-ri .2,41oxadiazol-3-ylmethyll-dimethyl-(3-phenoxy- propyQ-ammonium bromide

A mixture of 9-(3-dimethylaminomethyl-[1 ,2,4]oxadiazol-5-yl)-9H-xanthen-9-ol (74 mg, 0.291 mmol) and 1 -bromo-3-phenoxypropane (80 mg, 0.37 mmol) in isopropanol (3 ml) was refluxed for 18 h. The reaction mixture was concentrated in vacuo and purified by flash column chromatography eluting with 0-5% methanol-DCM to give the impure product as a white solid. Further purification by flash column chromatography eluting with DCM:ethanol:ammonia (0.88) 400:8:1 then 200:8:1 then 100:8:1 gave the title compound (20 mg) as a white solid. LC-MS (Method 4): Rt 7.75 min, m/z 458.2 [M] ⁺ . ¹ H NMR 400 MHz δ (ppm)(DMSO- d ₆ ): 2.19 (2 H, s), 3.13 (6 H, s), 3.40-3.48 (2 H, m), 3.91 (2 H, t, J = 5.89 Hz), 4.81 (2

H, s), 6.86-7.00 (3 H, m), 7.17-7.35 (6 H, m), 7.41-7.46 (2 H, m), 7.69 (2 H, dd, J = 7.88, 1.69 Hz), 7.74 (1 H, s).

Example 54

Dimethyl-[5-(9-methyl-9H-xanthen-9-yl)-[1,2,4]oxadiazol-3 -yImethyl]-(3-phenoxy- propyl)-ammonium bromide

a. g-Methyl-QH-xanthene-θ-carboxylic acid (2-chloro-1 -hydroxyimino-ethvD-amide

A mixture of 9-methyl-9H-xanthene-9-carboxylic acid (420 mg, 1.75 mmol) (N. Latif, N. Mishriky, Can. J. Chem., 1966, 44(11), 1271-4) and O-(7-azobenzotriazol-1 -yl)- 1 ,1 ,3,3-tetramethyluronium hexafluorophosphate (1.1 g, 2.8 mmol) and diisopropylethylamine (1 ml) in DMF (8 ml) was stirred at room temperature for 2 h. The reaction mixture was partitioned between water and ethyl acetate. The aqueous phase was extracted with ethyl acetate. The combined organic phases were dried (Na ₂ SO ₄ ) and concentrated in vacuo to leave a yellow oil. Purification by flash column chromatography on a silica isolute cartridge eluting with pentane, then 4:1-0:1 pentane-ether gave the title compound (534 mg, 92%) as a brown oil.

b. 3-Chloromethyl-5-(9-methyl-9H-xanthen-9-ylH1.2.41oxadiazole A solution of θ-methyl-θH-xanthene-θ-carboxylic acid (2-chloro-1 -hydroxyimino-ethyl)- amide (432 mg, 1.31 mmol) in acetic acid (10 ml) was heated in a microwave reactor at 15O ⁰ C for 30 min.. The reaction mixture was partitioned between water and ethyl

acetate. The aqueous phase was extracted with ethyl acetate. The combined organic phases were dried (Na ₂ SO ₄ ) and concentrated in vacuo to leave a colourless oil. Purification by flash column chromatography on a silica isolute cartridge eluting with 1:0-1 :1 pentane-ether gave the title compound (200 mg, 49%) as an off-white solid.

¹ H NMR 400 MHz δ (ppm)(CHCl ₃ -d): 2.15 (3 H, s), 4.62 (2 H ₁ s), 7.04-7.10 (4 H, m), 7.15

(2 H ₁ d, J = 8.92 Hz), 7.30 (2 H, ddd, J = 8.25, 6.13, 2.68 Hz).

c. Dimethyl-r5-(9-methyl-9H-xanthen-9-yl)-π ,2,41oxadiazol-3-ylmethvπ-amine A solution of 3-chloromethyl-5-(9H-xanthen-9-yl)-[1 ,2,4]oxadiazole (61 mg, 0.195 mmol) in THF (1 ml) was added dropwise to a stirred solution of dimethylamine in THF (2 M, 3 ml, 6 mmol) and the reaction mixture was stirred for 2 h. A further quantity of dimethylamine solution in THF (2 M, 5 ml, 10 mmol) was added and stirring was continued for 4 h. The reaction mixture was concentrated in vacuo to leave a yellow oil. Purification using a SCX cartridge gave the title compound as a yellow oil (53mg, 84%). LC-MS (Method 9): Rt 2.23 min, m/z 322.2 [MH] ⁺

d. Dimethyl-r5-(9-methyl-9H-xanthen-9-yl)-ri ,2,41oxadiazol-3-ylmethyl1-(3-phenoxy- propyP-ammonium; bromide

A mixture of 9 dimethyl-[5-(9-methyl-9H-xanthen-9-yl)-[1 ,2,4]oxadiazol-3-ylmethyl]-

amine (78 mg, 0.24 mmol) and 1-bromo-3-phenoxypropane (500 mg, 2.3 mmol) in acetonitrile (2 ml) and chloroform (2 ml) was heated at 50 ⁰ C for 3 days. The reaction mixture was purified by flash column chromatography eluting with 0-10% methanol in DCM to give the title compound (103 mg, 79%) as a white solid. LC-MS (Method 4): Rt 9.05 min, m/z 456.3 [M] ⁺ . ¹ H NMR 400 MHz δ (ppm)(DMSO- d ₆ ): 2.14 (3 H, s), 2.22-2.33 (2 H ₁ m), 3.22 (6 H, s), 3.53-3.60 (2 H, m), 4.00 (2 H, t, J = 5.88 Hz), 4.89 (2 H, s), 6.88-6.98 (3 H, m), 7.12 (2 H, ddd, J = 7.96, 7.14, 1.30 Hz), 7.20-7.23 (2 H, m), 7.28-7.40 (6 H, m).

Examples 55 and 56

[5-(Cyclohexyl-hydroxy-phenyl-methyl)-[1,3,4]oxadiazol-2- ylmethylH2-(3,4- dichloro-benzyloxy)-ethyl]-dimethyl-ammonium bromide enantiomers 1 and 2

a. Cvclohexyl-hvdroxy-phenyl-acetic acid N'-(2-dimethylaminoacetvπ-hydrazide 1 ,1'-Carbonyl diimidazole (1.53 g, 9.45 mmol) was added to a stirred suspension of cyclohexyl-hydroxy-phenyl-acetic acid (2.11 g, 9 mmol) in dry DCM at room temperature. A pale yellow solution formed. After stirring for 75 minutes a solution of dimethylamino-acetic acid hydrazide in dry DCM was added rapidly dropwise. After stirring at room temperature for 90 hours the reaction mixture was diluted with DCM and stirred with saturated sodium bicarbonate solution for 30 minutes. The mixture was filtered to remove a small amount of insoluble material and then the filtrate was partitioned. The organic layer was washed with saturated sodium bicarbonate solution, water, brine and then dried (MgSO ₄ ), filtered and evaporated in vacuo. Purification by chromatography using a silica isolute cartridge eluting with 0-7% methanol in DCM gave the title compound (2.1 g, 70%) as a white solid. ¹ H NMR, 400 MHz, DMSOd ₆ : δ 1.0-1.38 (6 H, m), 1.56-1.63 (2 H, m), 1.68-1.79 (2 H, m), 2.2 (6 H, s), 2.2-2.8 (1 H, m), 2.88 (2 H, s), 5.53 (1 H, s), 7.18-7.25 (1 H, m), 7.28-7.36 (2 H, m), 7.55-7.6 (2 H, m), 9.51 (2 H, brs).

b. Cyclohexyl-(5-climethylaminomethyl-ri ,3,41oxadiazol-2-yl)-phβnyl-methanol, Enantiomers 1 and 2

A mixture of cyclohexyl-hydroxy-phenyl-acetic acid N'-(2-dimethylaminoacetyl)- hydrazide (1.8 g, 5.4 mmol) and acetic anhydride (22.5 ml) was stirred and heated at 90 ⁰ C for 1 hour. The cooled reaction mixture was poured onto a mixture of ice and saturated sodium carbonate solution with stirring. It was then treated with saturated sodium bicarbonate solution until neutral. The mixture was extracted twice with DCM and the combined organic extracts were washed with saturated sodium bicarbonate solution (twice), brine, dried (MgSO ₄ ), filtered and evaporated in vacuo to give crude material. This was redissolved in DCM and washed three times with saturated sodium bicarbonate solution, brine, dried (MgSO ₄ ), filtered and evaporated in vacuo to give an orange-brown syrup. Purification by chromatography using a silica isolute cartridge eluting with 0-3 % methanol in DCM gave the title compound as a yellow syrup (0.85 g, 50%) after coevaporation with toluene.

LC-MS (Method 5): R, 2.36 min, m/z 316 [MH] ⁺ .

Cyclohexyl-(5-dimethylaminomethyl-[1 ,3,4]oxadiazol-2-yl)-phenyl-methanol (300 mg) was separated into its two enantiomers by HPLC with a Chiralpak IA column, eluting with 10% isopropanol in heptane containing 0.1% diethylamine, to give Enantiomer 1 (100 mg; R, 11.1 min) as a colourless gum, and, following further chiral HPLC purification, Enantiomer 2 (55 mg; R _t 15.4 min) also as a colourless gum.

c. r5-(SH-Cvdohexyl-hvdroxy-phenyl-methylH1 ,3,4]oxadiazol-2-ylmethylH2-(3,4- dichloro-benzyloxy)-ethvfl-dimethyl-ammonium bromide and r5-(RV(-cycloheχyl- hvdroxy-phenyl-methyD-π .Sλloxadiazol^-ylmethyll-^-Oλ-dichloro-benzyloxyl-ethvn- dimethyl-ammonium bromide,

A mixture of cyclohexyl-(5-dimethylaminomethyl-[1 ,3,4]oxadiazol-2-yl)-phenyl- methanol, enantiomer 1 as ^" obtained in the previous step (50 mg, 0.159 mmol) and 4-

(2-bromo-ethoxymethyl)-1 ,2-dichlorobenzene (225 mg, 0.795 mmol) in chloroform (0.9 ml) and acetonitrile (0.6 ml) was heated at 5O ⁰ C for 72 h. Purification by flash column chromatography (on a silica isolute cartridge) eluting with diethyl ether, dichloromethane, then 2-10% methanol-dichloromethane gave the first title compound as a white solid (49 mg, 58%).

LC-MS (Method 8): R, 9.13 min, m/z 518.2 [M] ⁺ .

¹ H NMR 400 MHz δ (ppm)(DMSO-d ₆ ): 0.91-1.25 (5 H, m), 1.31 (1 H, d, J = 12.44 Hz),

1.50-1.75 (4 H, m), 2.26 (1 H, t, J = 10.92 Hz), 3.14-3.22 (6 H, m), 3.67-3.72 (2 H, m),

3.92-3.97 (2 H, m), 4.53 (2 H, s), 5.06 (2 H, s), 6.42 (1 H, s), 7.25-7.30 (1 H, m), 7.32-

7.37 (3 H, m), 7.44-7.49 (2 H, m), 7.60-7.64 (2 H, m). ^{■ '} '

The second title compound was obtained in an analogous manner from cyclohexyl-(5- dimethylaminomethyl-[1 ,3,4]oxadiazol-2-yl)-phenyl-methanol, enantiomer 2 as obtained in the previous step. LC-MS (Method 8): R, 9.13 min, m/z 518.2 [M] ⁺ .

Examples 57- 71

The following examples were obtained from racemic cyclohexyl-(5- dimethylaminomethyl-f1 ,3,41oxadiazol-2-yl)-phenyl-methanol by methods analogous to those in Example 1 , with the exception of Example 71 which was prepared from (R)- cvclohexyl-(5-dimethylaminomethyl-n ,3,41oxadiazol-2-yl)-phenyl-methanol (obtained from (R)-cyclohexylmandelic acid) by methods analogous to those in Example 1.

Example 72

{2-[5-(Hydroxy-diphenyl-methyl)-isoxazol-3-yl]-ethyl}-dim ethyl-(3-phenoxy- propyl)-ammonium bromide

a. rδ-dHvdroxy-diphenyl-methvπ-isoxazol-S-yli-acetonitrile

Sodium cyanide (54 mg, 1.1 mmol) was added to a suspension of (3-bromomethyl- isoxazol-5-yl)-diphenyl-methanol (345 mg, 1.0 mmol) (prepared as described in Example I, Step d) in IMS (5 ml_) and the mixture was heated to 70 ⁰ C for 4.5 h. The solvent was evaporated in vacuo then the residue was taken up in ethyl acetate and water and extracted with ethyl acetate (x 2). The organic extracts were washed with brine then dried (MgSO ₄ ), filtered and concentrated in vacuo to afford the title compound (285 mg, 98 %) as a yellow oil. LCMS (Method 9): R _t 3.34 min, m/z 291.2 [M+HJ ⁺ . _;

b. r3-(2-Amino-ethyl)-isoxazol-5-yll-diphenyl-methanol

Borane dimethyl sulphide complex (1.79 mL, 2 M in THF, 3.57 mmol) was added dropwise to a solution of [5-(hydroxy-diphenyl-methyl)-isoxazol-3-yl]-acetonitrile (259 mg, 0.89 mmol) in THF (20 mL) at 55°C then the mixture was heated to reflux for 3 h. The reaction mixture was cooled to 0 ⁰ C before addition of MeOH (2 mL) and 1M HCI (1 mL). Stirring was continued for 30 mins at RT then the solvent was removed in vacuo. The residue was dissolved in DCM and saturated aqueous NaHCO ₃ solution and the phases were separated using a hydrophobic frit. The crude material was dissolved in DCM and purified by SCX cartridge to give the title compound (175 mg, 67 %) as a yellow foam. LCMS (Method 9): R _t 2.44 min, m/z 295.01 [M+H] ⁺ .

c. [3-(2-Dimethylamino-ethyl)-isoxazol-5-vn-diphenyl-methanol Formaldehyde solution (476 μL, 37% in water, 6.36 mmol) was added to a solution of [3-(2-amino-ethyl)-isoxazol-5-yl]-diphenyl-methanol (187 mg, 0.64 mmol) in 1 ,2-DCE (5 mL) followed by sodium triacetoxyborohydride (270 mg, 1.27 mmol). The reaction

mixture was stirred at RT for 1.75 h before treatment with a saturated solution of aqueous NaHCO ₃ . DCM was added after stirring for 15 min. then the phases were separated using a hydrophobic frit and the organics evaporated in vacuo. The crude material was dissolved in MeOH and purified by SCX cartridge. Further purification using CombiFlash Companion ® system over silica eluting with 0-10 % MeOH in DCM gave the title compound (110 mg, 45 %) as a colourless gum. LCMS (Method 9): R,2.26 min, m/z 323.31 [M+Hf.

d. {2-r5-(Hvdroxy-diphenyl-methylHsoxazol-3-vN-ethyl)-dimethyl- (3-phenoxy-propyl)- ammonium bromide

The title compound was prepared from [3-(2-dimethylamino-ethyl)-isoxazol-5-yl]- diphenyl-methanol and 3-phenoxypropyl bromide using a method analogous to that described for Example 1. Yield 45 mg, 42%. LCMS (Method 4): R, 8.21 min, m/z 457 [M] ⁺ .

¹ H NMR 400 MHz δ (ppm)(DMSO-d): 2.13-2.24 (2 H, m), 3.10-3.26 (8 H, m), 3.49-

3.57 (2 H, m), 3.66-3.73 (2 H, m), 4.00-4.07 (2 H, m), 6.28 (1 H, s), 6.92-6.98 (3 H, m), 7.09 (1 H, s), 7.23-7.38 (12 H, m).

Example 73

^-(Cyclohexyl-hydroxy-phenyl-methylJ-S-methyl-SH-irnidazo l-^ylmethylJ-dimethyl- (3-phenoxy-propyl)-ammonium bromide

a. 5-π ,3lDioxolan-2-yl-1-methyl-1 H-imidazole

A solution of 1 -methyl-1 H-imidazole-5-carboxaldehyde (3 g, 0.0272 mol), ethylene glycol (9 mL, 0.16 mol) and p-toluenesulfonic acid (465 mg, 0.0024 mol) in toluene (80 mL) was refluxed with azeotropic removal of water overnight. The reaction mixture was allowed to cool and washed with saturated aqueous sodium bicarbonate solution (40 mL). The aqueous phase was extracted with dichloromethane (60 mL). The

combined organic layers were dried (MgSO ₄ ), filtered and evaporated in vacuo to give the title compound (3.8 g, 90%), contaminated with a small amount (<5%) of the starting material.

b. Cyclohexyl-(5-π ,3ldioxolan-2-yl-1 -methyl-1 H-imidazol-2-vO-phenyl-methanol A solution of 5-[1 ,3]dioxolan-2-yl-1-methyl-1 H-imidazole (3.8 g, 0.0246 mol) in THF (125 ml_) was cooled to O ⁰ C under nitrogen. n-Butyllithium (2.5 M solution in hexanes, 12.8 ml_, 0.032 mol) was added and the reaction mixture allowed to stir at 0 ⁰ C for 40 minutes. A solution of cyclohexylphenyl ketone (5.55 g, 0.0295 mol) in THF (25 mL) was added and the reaction mixture allowed to warm to room temperature over 4 h, and then stirred overnight. The reaction mixture was quenched with saturated aqueous ammonium chloride solution and extracted with ether. The organic layer was dried (MgSO ₄ ), filtered and evaporated in vacuo. Purification by column chromatography eluting with 1 :0-9:1 DCM/MeOH gave the title compound (3.73 g, 44%). LC-MS (Method 5) R, 2.30 m/z 343 [MH] ⁺ .

c. 2-(Cvclohexyl-hvdroxy-phenyl-methyl)-3-methyl-3H-imidazole-4 -carbaldehvde A solution of cyclohexyl-(5-[1 ,3]dioxolan-2-yl-1 -methyl-1 H-imidazol-2-yl)-phenyl- methanol (3.7 g, 0.0108 mol) in THF (50 mL) was treated with 1 M aqueous HCI (50 mL) and heated at 6O ⁰ C overnight. The reaction mixture was evaporated in vacuo to remove THF. The remaining aqueous layer was neutralized with aqueous saturated sodium hydrogen carbonate. The resulting solid was collected by filtration and washed with water, then re-dissolved in DCM, dried (MgSO ₄ ), filtered and evaporated in vacuo to give the title compound (2.33 g, 72%). LC-MS (Method 5): R ₁ 3.55 m/z 299 [MH] ⁺ .

d. Cvclohexyl-fδ-dimethylaminomethyl-i -methyl-1 H-imidazol-2-vπ-phenyl-methanol A solution of 2-(cyclohexyl-hydroxy-phenyl-methyl)-3-methyl-3H-imidazole-4 - carbaidehyde (2.3 g, 0.00772 mol) in 1 ,2-dichloroethane (80 ml_) was treated with a solution of dimethylamine (4.63 mL of a 2.0 M solution in THF, 0.00926 mol) and stirred at room temperature for 2 h. Sodium triacetoxyborohydride (1.96 g, 0.00926 moles) was added and the reaction mixture was allowed to stir at room temperature overnight. An additional quantity of dimethylamine (0.6 equivalents) and sodium triacetoxyborohydride (0.6 equivalents) was added and the reaction stirred for a further 4 h. The reaction mixture was diluted with DCM and washed with saturated aqueous sodium hydrogen carbonate solution (60 mL). The organic layer was separated, dried over magnesium sulphate, filtered and evaporated in vacuo to give the crude product. Purification on a neutral alumina column, eluting with 0-25% methanol-dichloromethane gave the title compound (1.0 g, 40%). LC-MS (Method 5) R, 1.85 m/z 328 [MH] ⁺ .

The title compound was separated into its enantiomers by chiral HPLC using a 250 x 21.2 mm Chiralpak ® IA column packed with amylase tris(3,5- dimethylphenylcarbamate) immobilized on 5 μm silica gel. The column was eluted with 2% IPA in heptane buffered with 0.1 % diethylamine. Flow rate 1 ml/min. Wavelength, 220 nm.

1 ^st enantiomer R _t = 15.2 min; 2 ^nd enantiomer R _t = 18.2 min.

The second eluting enantiomer required further chiral purification using the conditions described above.

e. [2-(Cvclohexyl-hvdroxy-phenyl-methyl)-3-methyl-3H-imidazol-4 -ylmethvπ-dimethyl- (3-phenoxy-propyl)-ammonium bromide

To a suspension of cyclohexyl-(5-dimethylaminomethyl-1 -methyl-1 H-imidazol-2-yl)- phenyl-methanol, 1st enantiomer (30 mg, 0.092 mmol), in chloroform (0.5 mL) and acetonitrile (0.5 mL) was added 1 -bromo-3-phenoxypropane (0.162 mL, 0.92 mmol). The mixture was stirred at 50 ⁰ C overnight. The reaction mixture was evaporated and purified by column chromatography eluting with 0-10% methanol-DCM to give the first title compound (38 mg, 76%). LC-MS (Method 8) R, 7.37 m/z 462 [M] ⁺ .

¹ H NMR 400 MHz δ (ppm)(CHCI ₃ -d): 0.80-1.70 (9 H, m), 2.07 (1 H, d, J = 12.06 Hz), 2.25 (2 H, dd, J = 11.44, 5.86 Hz), 2.58 (1 H, t, J = 11 <78 Hz), 3.18 (3 H, s), 3.24 (3 H, 0 s), 3.38 (1 H, S) ₁ 3.57 (3 H, s), 3.71-3.79 (2 H, m), 4.05 (2 H, t, J = 5.33 Hz), 4.83 (1 H ₁ d, J = 14.66 Hz), 5.08 (1 H, d, J = 14.66 Hz), 6.83 (2 H, d, J = 8.11 Hz), 6.93-6.99 (1 H, m), 7.18-7.29 (8 H, m).

5 The following Examples were prepared by methods analogous to those described for Example 73 using enatiomer 1 of Example 73 Step d, except for Example 80, which used enantiomer 2 from Example 73 Step d.

75 [2-(Cyclohexyl- 7.30 ¹ H NMR δ (ppm) hydroxy-phenyl- 446 (Method (CHCI ₃ -d): 0.96-1.30 (5 methyl)-3-methyl- 8) H ₁ m), 1.38-1.52 (1 H,

3H-imidazol-4- m), 1.63 (2 H, s), 1.81 ylmethyl]-dimethyl- (1 H, d, J = 13.11 Hz),

(3-phenyl-propyl)- 2.03-2.13 (3 H, m), ammonium 2.59 (1 H, t, J = 11.77 bromide Hz), 2.65-2.76 (2 H, m), 3.16 (3 H, s), 3.18 (3 H, s), 3.41-3.61 (5 H, m), 4.79 (1 H, d, J = 14.63 Hz) ₁ 5.10 (1 H, d, J = 14.63 Hz), 7.11 (1 H, s), 7.13-7.33 (10 H, m).

76 [2-(Cyclohexyl- 7.62 ¹ H NMR δ (ppm) hydroxy-phenyl- 469 (Method (CHCI ₃ -d): 0.90-1.28 (6 methyl)-3-methyl- 8) H, m), 1.39-1.48 (1 H,

3H-imidazol-4- m), 1.52-1.94 (6 H, m), ylmethyl]-dimethyl- 2.11 (1 H, d, J = 12.07

(3-phenyl-butyl)- Hz), 2.55-2.71 (3 H, ammonium m), 3.11 _, (3 H, s), 3.16 bromide (3 H, S) ₁ 3.45-3.61 (5 H, m), 4.79 (1 H, d, J = 14.64 Hz), 5.06 (1 H ₁ d, J = 14.63 Hz), 7.10 (1 H, s), 7.12-7.31 (1O H, m).

77 (4-Cyano-benzyl)- 6.53 ¹ H NMR δ

[2-(cyclohexyl-hyd 443 (ppm)(DMSO-d ₆ ): 0.84- roxy-phenyl- (Method 8) 1.34 (6 H, m), 1.58- methyl)-3-methyl- 1.62 (2 H, m), 1.72 (1

3H-imidazol-4- H, d, J = 12.42 Hz),

Example 81 rS-fCyclohexyl-hvdroxy-phenyl-methvD-isoxazol-S-ylmethvn-dim ethyl-O- phenoxy-propyD-ammonium bromide

a. 3-(Tetrahvdro-pyran-2-yloxymethyl)-isoxazole-5-carboxylic acid methoxy-amide To a solution of λ/,0-dimethylhydroxyIamine hydrochloride propiolate (21.17854 g mg, 215.88.76 mmol) were dissolved in acetonitrilein methylene chloride (2165 ml_) at 0 ⁰ C was added a solution of AIMe ₃ (14.13 g2.0 N solution in toluene, 64.74.38 ml_, 8.76 mmol). The reaction was stirred at room temperature for 30 min. The reaction mixture was cooled to 0 ⁰ C and a solution of 3-(tetrahydro-pyran-2-yloxymethyl)- isoxazole-5-carboxylic acid ethyl ester (744 mg, 2.92 mmol) in methylene chloride (4 m!_) was added. Reaction was stirred at O ⁰ C for 1 h and quenched with 1 N HCI. and DMAP (527 mg, 4.32 mmol) was added. The nitro compound (7.65 g, 43.17 mmol) dissolved in acetonitrile (216 mL) was added dropwise over a period of 1 h at room temperature and reaction mixture stirred overnight. The solvent was concentrated in vacuo. The aqueous layer was extracted with ethyl acetate and washed with sat. NaHCO _{3 (aq} ), brine, dried (MgSO ₄ ), filtered and concentrated in vacuoPurification using CombiFlash Companion ® system over silica using a 0-20 % gave to give the title compound (3.0625 g mg, 2779 %) as a yellow oil. ¹ H NMR, 400 MHz, CDCI ₃ : δ 6.94 (1 H, s), 4.86 - 74.80 (1 H, d, J = 12 Hz), 4.75 - 4.71 (1 H, m), 4.71 -4.65 (1 H, d, J = 11.6 Hz), 3.91 - 3.84 (1 H, m), 3.82 (3 H, s), 3.60 - 3.53 (1 H, m), 3.38 (3 H, s), 1.92 - 1.51 (6 H, m).

b. Phenyl-r3-tetrahvdro-pyran-2-yloxy-2-yloxymethyl)isoxazol-5- vn-methanone Phenyl magnesium bromide solution (1.0 M in THF, 4.2 mL, 4.2 mmol) was added dropwise to a solution of 3-(tetrahydro-pyran-2-yloxymethyl)-isoxazole-5-carboxylic acid methoxy-amide (562 mg, 2.3 mmol) in THF (7 mL) at 0 ^oO C. The reaction was stirred for 2 h and quenched with sat. NH ₄ CI ( _aq) . The aqueous layer was extracted with ethyl acetate, dried (MgSO ₄ ), filtered and the solvent concentrated in vacuo. Purification using Biotage SP1 © system over silica using a 0-40% ethyl acetate/cyclohexane as eluent gave the title compound (449 mg, 68%) as an oil. ¹ H NMR, 400 MHz, CDCI ₃ : δ 8.17 - 8.10 (2H, d, J = 7.3 Hz), 7.70 - 77.63 (1 H, m), 7.58

7.51 (2 H ₁ 1, J = 7.2 Hz), 7.10 (1 H, s), 4.92 - 4.86 (1 H ₁ d, J = 12.9 Hz), 4.79 - 4.75 (1 H, m), 4.74 - 4.69 (1 H, d, J = 12.9 Hz), 3.93 - 3.85 (1 H, m), 3.63 - 3.54 (1 H, m), 1.91 - 1.71 (2 H, m) 1.70 - 1.50 (4 H, m).

c. Cvclohexyl-phenyl-rS-ftθtrahvdro-pyran^-yloxymethvD-isoxazo l-δ-vn-methanol Cyclohexylmagnesium bromide solution (2.0 M in diethyl ether, 2.68 ml_, 5.36 _. mmol) was added dropwise to a solution of phenyl-[3-tetrahydro-pyran-2-yloxy-2- yloxymethyl)isoxazol-5-yl]-methanone (439 mg, 1.53 mmol) in THF (5 mL) at O ⁰⁰ C. The reaction was warmed to room temperature and stirred overnight. The reaction was quenched with sat. NH ₄ CI( _aq) and the aqueous layer extracted with ethyl acetate, dried (MgSO ₄ ), filtered and the solvent concentrated in vacuo. Purification using Biotage SP1 ® system over silica using a 0-20% ethyl acetate/cyclohexane as eluent gave the title compound (270 mg, 48%) as an oil. ¹ H NMR, 300 MHz, CDCI ₃ : δ 7.59 - 7.52 (2 H ₁ d, J = 8.0 Hz), 7.39 - 77.30 (2 H, t, J = 7 Hz), 7.30 - 7.22 (1 H, m), 6.30 (1 H, s), 4.78 - 4.63 (2 H, m), 4.59 - 4.51 (1 H, d, J = 12.9 Hz), 3.91 - 3.79 (1 H, m), 3.59 - 3.48 (1 H, m), 2.44 (1 H, s) 2.41 - 2.29 (1 H, m), 1.90 - 1.45 (9 H, m), 1.40 - 0.94 (7 H, m).

d. Cvclohexyl-P-hydroxymethyl-isoxazol-S-vD-phenyl-methanol

Pyridinium tosylate (18 mg, 0.073 mmol) was added to a solution of cyclohexyl- phenyl-[3-(tetrahydro-pyran-2-yloxymethyl)-isoxazol-5-yl]-me thanol (270 mg, 0.73 mmol) in IMS (7 mL). The reaction mixture was heated to 55 ^o0 C for 5.5 h and concentrated in vacuo. Purification using Biotage SP1 ©system over silica using a 0- 30% ethyl acetate/cyclohexane as eluent gave the title compound (178 mg, 85%) as a white foam. LCMS (Method 5): R, 3.43 min, m/z 288 [M+2H] ⁺ .

e. O-Bromomethyl-isoxazol-δ-vD-cvclohexyl-phenyl-methanol

Carbon tetrabromide (187 mg, 0.56 mmol) was was added to a solution of the cyclohexyl-(3-hydroxymethyl-isoxazol-5-yl)-phenyl-methanol (135 mg, 0.47 mmol) in methylene chloride (5 ml_) at 0 ⁰ C followed by triphenylphosphine (147 mg, 0.56 mmol). The reaction mixture was stirred for 20 min and then at room temperature for 2 h. The solvent was concentrated in vacuo. Purification using Biotage SP1 system ® system over silica using a 0-30 % ethyl acetate/cyclohexane as eluent gave the title compound (149.4 mg, 91%) as a white solid. ¹ H NMR, 300 MHz, CDCI ₃ : δ 7.59 - 7.50 (2H, m), 7.40 - 77.31 (2 H, t, J = 6.9 Hz), 7.30 - 7.27 (1 H, m), 6.34 (1 H, s), 4.36 (2 H, s), 2.44 (1 H, s), 2.41 - 2.28 (1 H, m), 1.83 - 1.60 (3 H, m), 1.54 - 1.44 (1 H, m), 1.41 - 0.92 (6 H, m).

f. Cyclohexyl-O-dimethylaminomethyl-isoxazol-S-vD-phenyl-methan ol

A solution of the (3-bBromomethyl-isoxazol-5-yl)-cyclohexyl-phenyl-methanol (143 mg, 0.41 mmol) in THF (3 ml_) was added dropwise over 1 h to a solution of dimethylamine (2.0 M in THF, 0.82 ml_, 1.64 mmol). The reaction mixture was stirred at room temperature overnight. The reaction was concentrated in vacuo and the residue partitioned between ethyl acetate and sat. NaHCO _{3 {aq)} . The organic layer was separated and the aqueous layer extracted with ethyl acetate. The combined organics were combined, dried (MgSO ₄ ), filtered and concentrated in vacuo. Purification using Biotage SP1 system ® system over silica using a 0-10% methanol/methylene chloride as eluent gave the title compound (84 mg, 65%) as an oil. LCMS (Method 5): R, 2.52 min, m/z 315 [M+2H] ⁺ .

q. rS-fCvclohexyI-hvdroxy-phenvI-methvD-isoxazol-S-vImethyli-di methyl-O-phenoxy- propyD-ammonium bromide To a solution of cyclohexyl-(3-dDimethylaminomethyl-isoxazol-5-yl)-phenyl-met hanol (26 mg, 0.083 mmol) in chloroform (0.3 ml) and CH ₃ CN (0.2 ml) was added 3- phenoxypropyl bromide (0.091 ml_, 0.58 mmol). The mixture was heated at 50 ⁰⁰ C for 48 h. After this period the solvents were evaporated and the residue purified using Biotage SP1 system ® system over silica using a 0-10% methanol/methylene chloride as eluent to gave give the title compound (27 mg, 73%) as a white solid. LC- MS (Method 5): R ₁ 2.71 min, m/z 450 [M] ⁺ . ¹ H NMR, 300 MHz ₁ DMSOd ₆ : δ 7.58 - 7.49 (2 H, d, J = 7.7 Hz), 7.41 - 7.22 (4 H, m), 7.00-6.88 (3 H, m), 6.70 (1 H, s), 6.15 (1 H, s), 4.73 (2 H, s), 4.09 - 3.99 (2 H, m), 3.56 - 3.43 (2 H, sm), 3.12 (6 H, s), 2.34 - 2.17 (3 H, m), 1.75 - 1.51 (3 H, m), 1 .42 - 0.92 (8 H, sm).

BIOLOGICAL EXAMPLES

The inhibitory effects of compounds of the present invention at the M3 muscarinic receptor were determined by the following .binding assays:

Muscarinic Receptor Radioligand Binding Assays

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

The half-life of antagonists at each muscarinic receptor was measured using the alternative radioligand [ ³ H]-QNB and an adaptation of the above affinity assay. Antagonists were incubated for 3 hours at a concentration 10-fold higher than their Ki,

as determined with the [ ³ H]-QNB ligand, with membranes expressing the human muscarinic receptors. At the end of this time, [ ³ H]-QNB was added to a concentration 25-fold higher than its Kd for the receptor being studied and the incubation continued for various time periods from 15 minutes up to 180 minutes. Membranes and bound radioligand were then harvested by filtration and allowed to dry overnight. Scintillation fluid was then added and the bound radioligand counted using a Canberra Packard Topcount scintillation counter.

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

Alternatively,

Recombinant human M3 receptor was expressed in CHO-K1 cells. Cell membranes were prepared and binding of [3H]-N-methyl scopolamine ([3H]-NMS) and compounds was assessed by a scintillation proximity assay (SPA). The incubation time was 16 hours at room temperature in the presence of 1 % (v/v) DMSO. The assay was performed in white 96 well clear-bottomed NBS plates (Corning). Prior to the assay, the CHO cell membranes containing M3 receptor were coated onto SPA WGA (Wheat germ agglutinin) beads (GE Healthcare). Non specific binding was determined in the presence of 1 μM Atropine.

Radioactivity was measured on a Microbeta scintillation counter (PerkinElmer) using a 3H protocol with a 2 minutes per well read time. Compound inhibition of [3H]-NMS binding was determined typically using concentrations in the range 0.03 nM to 1 μM and expressed as percent inhibition relative to the plate specific radioligand binding for the plate. Concentration dependent inhibition of [3H]-NMS binding by compounds was expressed as plC50.

Binding data for Examples of the invention, where tested, are shown in the table below.

M3 Ki <5nM "+++"; 5-2OnM "++", >20nM "+"

All compounds tested exhibited potencies in the M3 binding assay of greater than

1 μM. In particular, Example 12 exhibited a Ki value of 0.2OnM and Example 18 exhibited a Ki value of 0.21 nM.

Assay for adrenergic 62 mediated cAMP production CELL PREPARATION

H292 cells are grown in RPMI (Roswell Park Memorial Institute) medium containing, 10% (v/v) FBS (foetal bovine serum) and 2 mM L-glutamine. Cells are grown in 225cm2 flasks containing 25 mL media in a humidified incubator at 37°C, 5% CO ₂ . Cells are harvested from the flask and passaged at a 1 in 10 dilution once per week. EXPERIMENTAL METHOD The media from flasks containing H292 cells is removed, rinsed with 1O mL

PBS (phosphate buffered saline) and replaced with 1O mL Accutase™ cell detachment solution. Flasks are incubated for 15 minutes in a humidified incubator at 37°C, 5% CO ₂ . The cell suspension is counted and the cells re-suspended in RPMI media (containing 10% (v/v) FBS and 2 mM L-glutamine) at 0.05 x 10 ⁶ 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 overnight in a humidified incubator at 37°C, 5% CO ₂ . The culture media is removed, washed twice with 100 μL assay buffer and replaced with 50 μL assay buffer. Cells are rested at room temperature for 20 minutes after which time 25 μL of rolipram (1.2 mM made up in assay buffer containing 2.4% (v/v) dimethylsulphoxide) is added. Cells are incubated with rolipram for 10 minutes after which time test compounds (made up as x4 concentrated stocks in assay buffer containing 4% (v/v) dimethylsulphoxide) are added and the cells are incubated for 10 minutes at room temperature. Final rolipram concentration in the assay is 300 μM and final vehicle concentration is 1.6% (v/v) dimethylsulphoxide. The reaction is stopped by removing supematants, washing once with 100 μL assay buffer and replacing with 50 μL lysis buffer. The cell monolayer is frozen at -80 ⁰ C for 30 minutes (or overnight).

AlphaScreen™ cAMP Detection

The concentration of cAMP (cyclic adenosine monophosphate) in the cell lysate is determined using the 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 (containing equal volumes of donor beads (pre-incubated with biotinylated cAMP in the dark for 30 minutes) and acceptor beads), is added to each well and the plate incubated at room temperature for 10 hours 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 (made up in lysis buffer in a 96-well tissure-culture-treated plate and frozen/thawed alongside the test samples) and detected using the same protocol. Concentration response curves for agonists are constructed to determine both the pEC ₅₀ and Intrinsic Activity. Intrinsic Activity is expressed as a fraction relative to the maximum activity determined for formoterol in each experiment.

Alternative assay for Adrenergic β2 mediated cAMP production

Cell preparation

H292 cells are grown in 225cm2 flasks incubator at 37°C, 5% CO ₂ in RPMI medium containing10% (v/v) FBS (foetal bovine serum) and 2 mM L-glutamine. Experimental Method

Adherent H292 cells re removed from tissue culture flasks by treatment with Accutase™ cell detachment solution for 15 minutes. Flasks are incubated for 15 minutes in a humidified incubator at 37°C, 5% CO ₂ . Detached cells are re-suspended in RPMI media (containing 10% (v/v) FBS and 2 mM L-glutamine) at 0.05 x 10 ⁶ 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 overnight in a humidified incubator at 37°C, 5% CO ₂ . 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 1OmM HEPES pH7.4 and 5 mM glucose). Cells are rested at room temperature for 20 minutes after which time 25 μL of rolipram (1.2 mM made up in assay buffer containing 2.4% (v/v) , dimethylsulphoxide) is added. Cells are incubated with rolipram for 10 minutes after which time test compounds are added and the cells are incubated for 60 minutes at room temperature. The final rolipram concentration in the assay is 300 μM and final vehicle concentration is 1.6% (v/v) dimethylsulphoxide. 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 ⁰ C for 30 minutes (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-welI white plate. 40 μl_ of mixed AlphaScreen™ detection beads pre-incubated with biotinylated cAMP, is added to each well and the plate incubated at room temperature for 10 hours 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 pEC ₅₀ and Intrinsic Activity. Intrinsic Activity is expressed as a fraction relative to the maximum activity determined for formoterol in each experiment.

Analysis of Inhibition of M3 Receptor Activation via Calcium Mobilization CHO cells expressing the human M3 receptor were seeded and incubated overnight in 96 well collagen coated plates (black-wall, clear bottom) at a density of 50000 / 75μl of medium in 3% serum. The following day, a calcium-sensitive dye (Molecular Devices, Cat # R8041 ) was prepared in HBSS buffer with the addition of 5mM probenecid (pH 7.4). An equal volume of the dye solution (75μl) was added to the cells and incubated for 45 minutes followed by addition of 50μl of muscarinic antagonists or vehicle. After a further 15 minutes the plate was read on a FLEXstation™ (excitation 488nm, emission 525nm) for 15 seconds to determine baseline fluorescence. The muscarinic agonist Carbachol was then added at an EC _8O concentration and the fluorescence measured for a further 60 seconds. The signal was calculated by subtracting the peak response from the mean of the baseline fluorescence in control wells in the absence of antagonist. The percentage of the maximum response in the presence of antagonist was then calculated in order to generate IC ₅₀ curves

Evaluation of potency and duration of action in Isolated Guinea Pig

Experiments were carried out at 37 ⁰ C in modified Krebs-Henseleit solution, (1 14mM

NaCI, 15mM NaHCO ₃ , 1 mM MgSO ₄ , 1.3mM CaCI ₂ , 4.7mM KCI, 11.5mM glucose and 1.2mM KH ₂ PO ₄ , pH 7.4) gassed with 95% O ₂ /5% CO ₂ . lndomethacin was added to a final concentration of 3μM

Tracheae were 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 were cut and suspended using cotton thread in 10ml water-jacketed organ baths and attached to a force transducer ensuring that the tissue is located between two platinum electrodes. Responses were recorded via a MPI OOW/Ackowledge data acquisition system connected to a PC. Tissues were equilibrated for one hour under a resting tone of 1 g and were then subjected to electrical field stimulation at a frequency of 80Hz with a pulse width of 0.1 ms, a unipolar pulse, triggered every 2 minutes. A "voltage-response" curve was generated for each tissue and a submaximal voltage then applied to every piece of tissue according to its own response to voltage. Tissues were washed with Krebs solution and allowed to stabilize under stimulation prior to addition of test compound. Concentration response curves were 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 was 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 EC ₅₀ calculated for each compound.

Onset time and duration of action studies were performed by adding the previously determined EC ₅₀ concentration of compound to EFS contracted tissues and the response allowed to plateau. The time taken to reach 50% of this response was determined to be the onset time. Tissues were 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. All compounds that were tested in this assay showed an EC ₅₀ value of <50nM. In particular, Example 2 exhibited an EC ₅₀ value of 2.OnM

Methacholine Induced Bronchoconstriction in vivo

Male Guinea pigs (Dunkin Hartley), weighing 500-60Og housed in groups of 5 were individually identified. Animals were allowed to acclimatize to their local surroundings for at least 5 days. Throughout this time and study time animals were allowed access

to water and food ad libitum.

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

Up to 72hrs post dosing guinea pigs were terminally anaesthetized with Urethane (250μg/ml, 2ml/kg). At the point of surgical anaesthesia, the jugular vein was cannulated with a portex i.v. cannula filled with heparinised phosphate buffered saline (hPBS) (10U/ml) for i.v. administration of methacholine. The trachea was exposed and cannulated with a rigid portex cannula and the oesophagus cannulated transorally with a flexible portex infant feeding tube. The spontaneously breathing animal was then connected to a pulmonary measurement system (EMMS, Hants, UK) consisting of a flow pneumotach and a pressure transducer. The tracheal cannula was attached to a pneumotach and the oesophageal cannula attached to a pressure transducer.

The oesophageal cannula was positioned to give a baseline resistance of between 0.1 and 0.2cmH20/ml/s. A 2 minute baseline reading was recorded before i.v. administration of methacholine (up to 30μg/kg, 0.5ml/kg). A 2 minute recording of the induced constriction was 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 were used to analyse the bronchoprotective effects of test compounds.

Inhibition of pilocarpine induced salivation by i.n. administered compounds Guinea pigs (450-55Og) supplied by Harlan UK or David Hall, Staffs UK and acclimatised to the in-house facilities for a minimum of three days before use. Guinea pigs were randomly assigned into treatment groups and weighed. Each animal was lightly anaesthetised (4% Halothane) and administered compound or vehicle intranasally (0.5ml/kg) at up to 24 hours before challenge with pilocarpine. At the test time point, guinea pigs were terminally anaesthetised with urethane (25% solution in H20, 1.5g/kg). Once sufficient anaesthesia had developed (absence of toe pinch reflex) each animal had an absorbent pad placed in the mouth for 5 minutes to dry residual saliva, this pad was removed and replaced with a new pre-weighed pad for 5 minutes to establish a reading of baseline saliva production. At the end of this 5 minute period the pad was removed and weighed. A new pre-weighed pad was inserted into the mouth before each animal received s.c. pilocarpine administered

under the skin at the back of the neck (0.6mg/kg @ 2ml/kg). The pad was removed, weighed and replaced with a new pre-weighed pad every 5 minutes up to 15 minutes.

Saliva production was calculated by subtracting the pre-weighed weight of the pad from each 5 minute period post weighed pad and these numbers added together to produce an accumulation of saliva over 15 minutes. Each 5 minute period could be analysed in addition to the whole 15 minute recording period. Baseline production of saliva was assumed to be constant and multiplied by three to produce a reading for baseline saliva production over 15 minutes.

Inhibition of saliva produced by the compound could be calculated by using the following equation:

(1 -(Test-baseline)/(Veh-baseline)) ^* 100.