AZONIATRICYCLO [3.3.1.0] NONANE DERIVATIVES AS MUSCARINIC RECEPTOR ANTAGONISTS

Field of the Invention

This present invention generally relates to muscarinic receptor antagonists, which are useful, among other uses, for the treatment of various diseases of the respiratory, 5 urinary and gastrointestinal systems mediated through muscarinic receptors. The invention also relates to the process for the preparation of disclosed compounds, pharmaceutical compositions containing the disclosed compounds, and the methods for treating diseases mediated through muscarinic receptors. Also provided herein are pharmaceutical compositions comprising one or more muscarinic receptor antagonists and 10 at least one other active ingredients, which may be selected from, for example, histamine antagonists, corticosteroids, beta agonists, leukotriene antagonists, EGFR (epidermal growth factor receptor) kinase inhibitors, PAF (platelet activating factor) antagonists, 5- lipoxygenase inhibitors, chemokine inhibitors, PDE-4 inhibitors or p38 MAP kinase inhibitors.

15 Background of the Invention

Physiological effects elicited by the neurotransmitter acetylcholine are mediated through its interaction with two major classes of acetylcholine receptors - the nicotinic and muscarinic acetylcholine receptors. Muscarinic receptors belong to the superfamily of G-protein coupled receptors and five molecularly distinct subtypes are known to exist (M ₁ , 20 M ₂ , M ₃ , M ₄ and M ₅ ).

These receptors are widely distributed on multiple organs and tissues and are critical to the maintenance of central and peripheral cholinergic neurotransmission. The regional distribution of these receptor sub-types in the brain and other organs has been documented (for example, the Mi subtype is located primarily in neuronal tissues such as 25 cereberal cortex and autonomic ganglia, the M ₂ subtype is present mainly in the heart and bladder smooth muscle and the M ₃ subtype is located predominantly on smooth muscle and salivary glands {Nature, 323, (1986), 411; Science, 232, (1987), 527).

A review in Curr. Opin. Chem. Biol., 3, (1999), 426 as well as in Trends in Pharmacol. Sci., 22, (2001), 409 by Eglen et. al., describes the biological potentials of 30 modulating muscarinic receptor subtypes by ligands in different disease conditions, such

as Alzheimer's disease, pain, urinary disease condition, chronic obstructive pulmonary disease, and the like.

The pharmacological and medical aspects of the muscarinic class of acetylcholine agonists and antagonists are presented in a review in Molecules, 6, (2001), 142. Birdsall et. al., in Trends in Pharmacol. ScL, 22, (2001), 215 has also summarized the recent developments on the role of different muscarinic receptor subtypes using different muscarinic receptor of knock-out mice.

Almost all the smooth muscles express a mixed population of M ₂ and M ₃ receptors. Although the M ₂ - receptors are the predominant cholinoreceptors, the smaller population of M ₃ - receptors appear to be the most functionally important as they mediate the direct contraction of these smooth muscles. Muscarinic receptor antagonists are known to be useful for treating various medical conditions associated with improper smooth muscle function, such as overactive bladder syndrome, irritable bowel syndrome and chronic obstructive pulmonary disease. However, the therapeutic utility of antimuscarinics has been limited by poor tolerability as a result of treatment related, frequent systemic adverse events such as dry mouth, constipation, blurred vision, headache, somnolence and tachycardia. Thus, there exists a need for novel muscarinic receptor antagonists that demonstrate target organ selectivity.

WO 2004/005252 discloses azabicyclo derivatives described as muscarinic receptor antagonists. WO 2004/004629, WO 2004/052857, WO 2004/067510, WO 2004/014853, WO 2004/014363, and WO 2006/054162 disclose 3,6-disubstituted azabicyclo [3.1.0] hexane derivatives as useful muscarinic receptor antagonists. WO 2004/056811 discloses flaxavate derivatives as muscarinic receptor antagonists. WO 2004/056810 discloses xanthene derivatives as muscarinic receptor antagonists. WO 2004/056767 discloses l-substituted-3 -pyrrolidine derivatives as muscarinic receptor antagonists. WO 99/14200, U.S. Patent No. 6,200,991, and WO 00/56718 disclose heterocycle derivatives as muscarinic receptor antagonists. WO 2004/089363, WO 2004/089898, WO 2004/069835, WO 2004/089900 and WO 2004/089364 disclose substituted azabicyclohexane derivatives as muscarinic receptor antagonists. WO 2006/018708 discloses pyrrolidine derivatives as muscarinic receptor antagonists. WO 2006/035303 discloses azabicyclo derivatives as muscarinic receptor antagonists. U.S.

Publication No. 2004/0242622 discloses azabicycloalkane compounds as muscarinic receptor antagonists. U.S. Patent No. 5,610,163 discloses esters of thienyl carboxylic acid and their quaternization products which are anticholingeric agents. U.S. Publication No. 2003/0191316 discloses ester derivatives as muscarinic M3 receptor antagonist. WO 2006/025324 discloses tropane compounds for preventing and treating muscarinic receptor interposition property diseases such as chronic obstructive pulmonary disease and asthma.

/. Med. Chem., 44, (2002), 984 describes cyclohexylmethylpiperidinyl- triphenylpropioamide derivatives as selective M ₃ antagonist discriminating against the other receptor subtypes. /. Med. Chem., 36, (1993), 610 describes the synthesis and antimuscarinic activity of some l-cycloalkyl-l-hydroxy-l-phenyl-3-(4-substituted piperazinyl)-2-propanones and related compounds. /. Med. Chem., 34, (1991), 3065 describes analogues of oxybutynin, synthesis and antimuscarinic activity of some substituted 7-amino-l-hydroxy-5-heptyn-2-ones and related compounds. Bio-Org. Med. Chem. Lett., 15_, (2005), 2093 describes synthesis and activity of analogues of oxybutynin and tolterodine, Chem. Pharm. Bull., 53, (4): (2005), 437 discloses thiazole carboxamide derivatives.

Summary of the Invention

According to one aspect, there are provided muscarinic receptor antagonists, which can be effective therapeutic or prophylactic agents for the treatment of various diseases of the respiratory, urinary and gastrointestinal systems. Also provided are processes for synthesizing such compounds.

According to another aspect, the enantiomers, diastereomers, N-oxides, polymorphs and pharmaceutically acceptable solvates of these compounds as well as their metabolites are also provided. According to yet another aspect, pharmaceutical compositions containing such compounds are provided together with acceptable carriers, excipients or diluents which can be useful for the treatment of various diseases of the respiratory, urinary and gastrointestinal systems.

Also provided herein are pharmaceutical compositions comprising one or more muscarinic receptor antagonists and at least one other active ingredients include, but are

not limited to, histamine antagonists, corticosteroids, beta agonists, leukotriene antagonists, EGFR kinase inhibitors, PAF antagonists, 5 -lipoxygenase inhibitors, chemokine inhibitors, PDE-4 inhibitors or p38 MAP kinase inhibitors.

Other aspects will be set forth in the description which follows, and in part will be apparent from the description or may be learnt by the practice of the invention.

Detailed Description of the Invention

The present invention, relates to compounds having the structure of Formula I:

wherein

Ri is thienyl or cycloalkyl which is further substituted with difluoro on same carbon atom;

R ₂ is aralkyl, cycloalkyl, heterocyclylalkyl, heterocyclyl, heteroaryl or heteroarylalkyl which may further be substituted with methyl, ethyl, halogen, hydroxy or amino; with the proviso that when Ri is thienyl then R ₂ is aralkyl ; and when R ₁ is cycloalkyl which is further substituted with difluoro on same carbon atom, then R ₂ is aralkyl, cycloalkyl, heterocyclylalkyl, heterocyclyl, heteroaryl or heteroarylalkyl.

Z ^" is an anion selected from bromide, chloride, iodide, tartrate, sulphate, phosphate, nitrate, carbonate, fumarate, glutamate, citrate, methanesulphonate, benzenesulphonate, maleate and succinate. In one embodiment, the invention encompasses compounds of Formula I, which may include, but not limited to the following, for example:

9-(4-Fluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-meth yl-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 1);

7-[2-Hydroxy(di-2-thienyl)acetoxy]-9-methyl-9-(4-methylbenzy l)-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 2);

9-(4-Bromobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-methy l-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 3);

9-Benzyl-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-methyl-3-oxa-9 - azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 4);

9-(2,5-Difluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9- methyl-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 5); 9-(3-Bromobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-methy l-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 6);

9-(l,3-Benzodioxol-5-ylmethyl)-7-[2-hydroxy(di-2-thienyl)ace toxy]-9-methyl-3- oxa-9-azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 7);

9-(3,5-Difluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9- methyl-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 8);

9-(3-Fluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-meth yl-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 9);

9-(2-Fluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-meth yl-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 10); 9-(3,4-Difluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9- methyl-3-oxa-9- azonia-tricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 11) or

9-(2,5-Dibromobenzyl)-7-[2- hydroxy(di-2-thienyl)acetoxy]-9-methyl-3-oxa-9- azonia-tricyclo[3.3.1.0 ² ' ⁴ ] nonane bromide (Compound No. 12).

In another embodiment, there is provided herein a pharmaceutical composition comprising therapeutically effective amount of compound of Formula I described herein together with one or more pharmaceutically acceptable carrier(s), excipients(s) or diluent(s).

In yet another embodiment, there is provided a method for treatment or prophylaxis of a mammal suffering from a disease or disorder of the respiratory, urinary and gastrointestinal systems, wherein the disease or disorder is mediated through muscarinic receptors. The method includes administration of at least one compound having the structure of Formula I.

In yet another embodiment, there is provided a method for treatment or prophylaxis of a mammal suffering from a disease or disorder of the respiratory system such as bronchial asthma, chronic obstructive pulmonary disorders (COPD), pulmonary fibrosis, and the like; urinary system which induce such urinary disorders as urinary incontinence, lower urinary tract symptoms (LUTS), etc. and gastrointestinal system such as irritable bowel syndrome, obesity, diabetes and gastrointestinal hyperkinesis with any

one of the compounds as described above, wherein the disease or disorder is associated with muscarinic receptors.

In yet another embodiment, there is provided a pharmaceutical composition comprising one or more muscarinic receptor antagonist compound having the structure of Formula I as defined above and at least one other active ingredients include, but are not limited to, histamine antagonists, corticosteroids, beta agonists, leukotriene antagonists, EGFR kinase inhibitors, PAF antagonists, 5-lipoxygenase inhibitors, chemokine inhibitors, PDE-4 inhibitors or p-38 MAP kinase inhibitors.

In yet another embodiment, there are provided processes for preparing the compounds as described herein.

The following definitions apply to terms as used herein:

The term "alkyl," unless otherwise specified, refers to a monoradical branched or unbranched saturated hydrocarbon chain having from 1 to 20 carbon atoms. Alkyl groups can be optionally interrupted by atom(s) or group(s) independently selected from oxygen, sulfur, a phenylene, sulphinyl, sulphonyl group or -NR _α -, wherein R _α can be hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aryl, acyl, aralkyl, -C(=O)OR _λ , SO _m R _ψ or -C(=O)NRλR _π . This term can be exemplified by groups such as methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, t-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, n- decyl, tetradecyl, and the like. Alkyl groups may be substituted further with one or more substituents selected from alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto, oxo, thiocarbonyl, sunstituted thiocarbonyl, carboxy, carboxyalkyl, aryl, heterocyclyl, heteroaryl, (heterocyclyl)alkyl, cycloalkoxy, -NR _λ C(=O)OR _λ , COOR _λ, -CH=N-O(Ci_ ealkyl), -CH=N-NH(Ci_ ₆ alkyl), -CH=N-NH(Ci_ ₆ alkyl)-Ci_ ₆ alkyl, arylthio, thiol, alkylthio, aryloxy, alkoxyamino, nitro, aminosulfonyl, aminocarbonylamino, -NHC(=O)Rλ, -NRλR _π , -C(=O)NR _λ R _π , -NR _λ C(=O)NR _λ R _π , -C(=O)heteroaryl, C(=O)heterocyclyl, -O-C(=O)NR _λ R _π {wherein Rχ and R _π are independently selected from hydrogen, halogen, hydroxy, alkyl, alkenyl, alkynyl, alkenyl, alkoxy, cycloalkyl, cycloalkenyl, aryl, aralkyl, heterocyclyl, heteroaryl, heterocyclylalkyl, heteroarylalkyl, carboxy or Rx and R _π may also together join to form a heterocyclyl or heteroaryl ring} or -SO _m R _ψ (wherein m is an integer from 0-2 and R _ψ is hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aralkyl, aryl, heterocyclyl,

heteroaryl, heteroarylalkyl or heterocyclylalkyl). Unless otherwise constrained by the definition, alkyl substituents may be further substituted by 1-3 substituents selected from alkyl, alkenyl, alkynyl, carboxy, -NR _λ R _π , -C(=0)NR _λ R _π , -0C(=0)NR _λ R _π , -NRλC(=O)NRλR _π , hydroxy, alkoxy, halogen, CF3, cyano, and -S0mR _ψ ; or an alkyl group also may be interrupted by 1-5 atoms of groups independently selected from oxygen, sulfur or -NR _α - (wherein R _α , Rj ₁ , R _π , m and R _ψ are the same as defined earlier). Unless otherwise constrained by the definition, all substituents may be substituted further by 1-3 substituents selected from alkyl, alkenyl, alkynyl, carboxy, carboxyalkyl, -NR _λ R _π , -C(=0)NR _λ R _π , -0-C(=0)NR _λ R _π , hydroxy, alkoxy, halogen, CF ₃ , cyano, and -SO _m R _ψ (wherein Rj ₁ , R _π , m and R _ψ are the same as defined earlier); or an alkyl group as defined above that has both substituents as defined above and is also interrupted by 1-5 atoms or groups as defined above.

The term "alkylene," as used herein, refers to a diradical branched or unbranched saturated hydrocarbon chain having from 1 to 6 carbon atoms and one or more hydrogen can optionally be substituted with alkyl, hydroxy, halogen or oximes. This term can be exemplified by groups such as methylene, ethylene, propylene isomers (e.g., -CH ₂ CH ₂ CH ₂ and -CH(CH ₃ )CH ₂ ) and the like. Alkylene may further be substituted with one or more substituents such as alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, acyl, acylamino, acyloxy, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, oxo, thiocarbonyl, carboxy, arylthio, thiol, alkylthio, aryloxy, heteroaryloxy, aminosulfonyl, -COOR _ψ , -NHC(=O)Rλ, -NR _λ R _π , -C(=O)NR _λ R _π , -NHC(=O)NR _λ R _π , -C(=O)heteroaryl, C(=O)heterocyclyl, -O- C(=O)NRλR _π , nitro, -S(O) _m Rλ (wherein Rj ₁ , R _π , m and R _ψ are the same as defined earlier). Unless otherwise constrained by the definition, all substituents may be further substituted by 1-3 substituents chosen from alkyl, alkenyl, alkynyl, carboxy, -COOR _ψ , -NRλR _π , -C(=O)NR _λ R _π , -OC(=O)NR _λ R _π , -NHC(=O)NR _λ R _π , hydroxy, alkoxy, halogen, CF ₃ , cyano, and -S(O) _m R _ψ (wherein Rj ₁ , R _π , m and R _ψ are the same as defined earlier). Alkylene can also be optionally interrupted by 1-5 atoms of groups independently chosen from oxygen, sulfur and -NR _α (wherein R _α is the same as defined earlier). Unless otherwise constrained by the definition, all substituents may be further substituted by 1-3 substituents selected from hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, acyl, aralkyl, alkoxy, hydroxy, carboxy, -C(=O)OR _ψ , halogen, CF ₃ , cyano, -NR _λ R _π , -S(O) _m R _ψ , -C(=O)NR _λ R _π ,

-OC(=O)NR _λ R _π , -CONH-, -C=O or -C=NOH (wherein R _λ , R _π , m and R _ψ are the same as defined earlier).

The term "alkenyl," unless otherwise specified, refers to a monoradical of a branched or unbranched unsaturated hydrocarbon group having from 2 to 20 carbon atoms with cis, trans or geminal geometry. Alkenyl groups can be optionally interrupted by atom(s) or group(s) independently chosen from oxygen, sulfur, phenylene, sulphinyl, sulphonyl and -NR _α - (wherein R _α is the same as defined earlier). In the event that alkenyl is attached to a heteroatom, the double bond cannot be alpha to the heteroatom. Alkenyl groups may be substituted further with one or more substituents selected from alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, -NHC(=0)R _λ , -NR _λ R _π , -C(=0)NR _λ R _π , -NHC(=0)NR _λ R _π , -0-C(=0)NR _λ R _π , alkoxycarbonylamino, azido, cyano, halogen, hydroxy, oxo, keto, carboxyalkyl, thiocarbonyl, carboxy, arylthio, thiol, alkylthio, aryl, aralkyl, aryloxy, heterocyclyl, heteroaryl, heterocyclyl alkyl, heteroaryl alkyl, aminosulfonyl, aminocarbonylamino, alkoxyamino, hydroxyamino, alkoxyamino, nitro or SO _m R _ψ (wherein Rj ₁ , R _π , m and R _ψ are as defined earlier). Unless otherwise constrained by the definition, alkenyl substituents optionally may be substituted further by 1-3 substituents selected from alkyl, alkenyl, alkynyl, carboxy, hydroxy, alkoxy, halogen, -CF ₃ , cyano, -NRλR _π , -C(=O)NRλR _π , -O-C(=O)NRλR _π and -SO _m R _ψ (wherein Rj ₁ , R _π , m and R _ψ are as defined earlier). Groups, such as ethenyl or vinyl (CH=CH ₂ ), 1-propylene or allyl (-CH ₂ CH=CH ₂ ), iso-propylene (-C(CH ₃ )=CH ₂ ), bicyclo[2.2.1]heptene, and the like, exemplify this term.

The term "alkynyl," unless otherwise specified, refers to a monoradical of an unsaturated hydrocarbon, having from 2 to 20 carbon atoms. Alkynyl groups can be optionally interrupted by atom(s) or group(s) independently chosen from oxygen, sulfur, phenylene, sulphinyl, sulphonyl and -NR _α - (wherein R _α is the same as defined earlier). In the event that alkynyl groups are attached to a heteroatom, the triple bond cannot be alpha to the heteroatom. Alkynyl groups may be substituted further with one or more substituents selected from alkyl, alkenyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, keto, oxo, thiocarbonyl, carboxy, carboxyalkyl, arylthio, thiol, alkylthio, aryl, aralkyl, aryloxy, aminosulfonyl, aminocarbonylamino, hydroxyamino, alkoxyamino, nitro, heterocyclyl,

heteroaryl, heterocyclylalkyl, heteroarylalkyl, -NHC(=0)R _λ , -NR _λ R _π , -NHC(=0)NR _λ R _π , -C(=0)NR _λ R _π , -0-C(=0)NR _λ R _π or -SO _m R _ψ (wherein R _λ , R _π , m and R _ψ are the same as defined earlier). Unless otherwise constrained by the definition, alkynyl substituents optionally may be substituted further by 1-3 substituents selected from alkyl, alkenyl, alkynyl, carboxy, carboxyalkyl, hydroxy, alkoxy, halogen, CF ₃ , -NRj ₁ R ₁₁ , -C(=O)NR _λ R _π , -NHC(=0)NR _λ R _π , -C(=0)NR _λ R _π , cyano or -SO _m R _ψ (wherein R _λ , R _π , m and R _ψ are the same as defined earlier).

The term "cycloalkyl," unless otherwise specified, refers to cyclic alkyl groups of from 3 to 20 carbon atoms having a single cyclic ring or multiple condensed rings, which may optionally contain one or more olefinic bonds, unless otherwise constrained by the definition. Such cycloalkyl groups can include, for example, single ring structures, including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopentenyl, and the like or polycyclic ring structures such as, adamantyl, tricyclo[3.3.1.1]decane, bicyclo[2.2.2]octane, bicyclo[4.4.0]decane, bicyclo- [4.3.0]nonane, bicyclo[3.3.0]octane, bicyclo[2.2.1]heptane and the like, or cyclic alkyl groups to which is fused an aryl group, for example, indane, and the like. Spiro and fused ring structures can also be included. Cycloalkyl groups may be substituted further with one or more substituents selected from alkyl, alkenyl, alkynyl, alkoxy, cycloalkyl, cycloalkenyl, acyl, acylamino, acyloxy, alkoxycarbonylamino, azido, cyano, halogen, hydroxy, oxo, thiocarbonyl, carboxy, carboxyalkyl, arylthio, thiol, alkylthio, aryl, aralkyl, aryloxy, aminosulfonyl, aminocarbonylamino, -NRλR _π , -NHC(=O)NRλR _π , -NHC(=O)Rλ, -C(=O)NR _λ R _π , -O-C(=O)NR _λ R _π , -OC(=O)R _λ , -NR _λ C(=O)OR _λ , mercapto, haloalkyl, thioalkyl, -COOR _ψ , -COONHR _λ , -COR _λ , -NHSO ₂ R _λ or SO ₂ NHR _λ nitro, heterocyclyl, heteroaryl, heterocyclylalkyl, heteroarylalkyl or SO _m R _ψ (wherein Rj ₁ , R _π , m and R _ψ are the same as defined earlier). Unless otherwise constrained by the definition, cycloalkyl substituents optionally may be substituted further by 1-3 substituents selected from alkyl, alkenyl, alkynyl, carboxy, hydroxy, alkoxy, halogen, CF ₃ , -NRλR _π , -C(=O)NRλR _π , -NHC(=O)NR _λ R _π ,-OC(=O)NR _λ R _π , cyano or -SO _m R _ψ (wherein R _λ , R _π , m and R _ψ are the same as defined earlier). "Cycloalkylalkyl" refers to alkyl-cycloalkyl group linked through alkyl portion, wherein the alkyl and cycloalkyl are the same as defined earlier.

The term "alkoxy" denotes the group O-alkyl, wherein alkyl is the same as defined above.

The term "aryl," unless otherwise specified, refers to aromatic system having 6 to 14 carbon atoms, wherein the ring system can be mono-, bi- or tricyclic and are carbocyclic aromatic groups. For example, aryl groups include, but are not limited to, phenyl, biphenyl, anthryl or naphthyl ring and the like, optionally substituted with 1 to 3 substituents selected from halogen (e.g., F, Cl, Br, I), hydroxy, alkyl, alkenyl, alkynyl, cycloalkyl, alkoxy, acyl, aryloxy, CF ₃ , cyano, nitro, -CHO, OCF ₃ , -SCF ₃ , C00R _ψ , NHC(=0)R _λ , -NR _λ R _π , C(=N0H)NH ₂ , -C(=0)NR _λ R _π , -NR _λ C(=0)NR _λ R _π , NR _λ C(=0)0R _λ , -O-C(=O)NRλR _π , -S0mR _ψ , carboxy, heterocyclyl, heteroaryl, heterocyclylalkyl, heteroarylalkyl, acylamino, thiocarbonyl, substituted thiocarbonyl, amino carbonyl amino, mercapto, haloalkyl, optionally substituted aryl, optionally substituted heterocyclylalkyl, thioalkyl, -C0NHR _π , -OCOR _π , -COR _π , -NHSO ₂ R ₁₁ or -SO ₂ NHR ₁₁ (wherein R _λ , R _π , m and R _ψ are the same as defined earlier). Aryl groups optionally may be fused with a cycloalkyl group or a heteroaryl group, wherein the cycloalkyl group may optionally contain heteroatoms selected from O, N or S. Groups such as phenyl, naphthyl, anthryl, biphenyl, and the like exemplify this term.

The term "aralkyl," unless otherwise specified, refers to alkyl-aryl linked through an alkyl portion (wherein alkyl is as defined above) and the alkyl portion contains 1-6 carbon atoms and aryl is as defined below. Examples of aralkyl groups include benzyl, ethylphenyl, propylphenyl, naphthylmethyl and the like.

The term "aryloxy" denotes the group O-aryl, wherein aryl is as defined above. The term "carboxy," as defined herein, refers to -C(=0)0H.

The term "heteroaryl," unless otherwise specified, refers to an aromatic monocyclic, bicyclic or a tricyclic ring system (they can be fused, spiro or bridged) containing 1-8 heteroatom(s) independently selected from N, O or S optionally substituted with 1 to 4 substituent(s) selected from halogen (e.g., F, Cl, Br, I), hydroxy, alkyl, alkenyl, alkynyl, cycloalkyl, acyl, acylamino, thiocarbonyl, haloalkyl, thioalkyl, substituted thiocarbonyl, thioacyl, oxo, -CHO, -OCF ₃ , -CF ₃ , -SCF ₃ , carboxy, aryl, alkoxy, alkoxyamino, aralkyl, cyano, nitro, heterocyclyl, heteroaryl, -NRj ₁ R ₁₁ , CH=NOH,

-(CH ₂ ) _w C(=O)R _η ,COOR _λ {wherein w is an integer from 0-4 and R _η is hydrogen, hydroxy, OR _λ , NR _λ R _π , -NH0R _ω or -NHOH }, -C(=0)NR _λ R _π , -NR _λ C(=0)0R _λ , -NR _λ C(=O)NR _λ R _π , -SO _m R _ψ , -0-C(=0)NR _λ R _π , -O-C(=O)R _λ , -C00NHR _λ , -COR _λ , -NHSO ₂ R _λ or SO ₂ NHR _λ or -O-C(=O)OR _λ (wherein m, R _ψ , Rj ₁ and R _π are as defined earlier and R _ω is alkyl, cycloalkyl, aryl, heteroaryl, heterocyclyl, heteroarylalkyl or heterocyclylalkyl). Unless otherwise constrained by the definition, the substituents are attached to a ring atom, i.e., carbon or heteroatom in the ring. Examples of heteroaryl groups includes but are not limited to are benzimidazolyl, 1,4-benzodioxanyl, 1,3-benzodioxolyl, benzoxazolyl, benzothiazolyl, benzothienyl, benzo- triazolyl, dihydroimidazolyl, dihydropyranyl, dihydrofuranyl, dioxanyl, dioxolanyl, furyl, homopiperidinyl, imidazolyl, imidazolinyl, imidazolidinyl, indolinyl, indolyl, isoquinolinyl, isothiazolidinyl, isothiazolyl, isoxazolidinyl, isoxazolyl, morpholinyl, napthyridinyl, oxazolidinyl, oxazolyl, piperazinyl, piperidinyl, purinyl, pyrazinyl, pyrazolinyl, pyridinyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, pyrrolyl, pyrrolopyridinyl, imidazolpyridinyl, quinolinyl, tetrahydrofuranyl, quinozinyl, quinolizinyl, 6H-pyrido-[l,2-α]pyrimidinyl, tetrahydropyranyl, thiazolidinyl, thiazolyl, thienyl, pyridazinyl, carbazolyl, isobenzofuranyl, thianthrene, triazinyl, furanyl, benzofuranyl, tetrazolyl, quinazolinyl, benzoxazinonyl, benzothiazinonyl, benzimidazolone, pyrazolone, xanthene and the like.

The term "halogen or halo" refers to fluorine, chlorine, bromine or iodine. The term "haloalkyl" refers to alkyl of which one or more hydrogen(s) is/are replaced by halogen.

The term "heterocyclyl," unless otherwise specified, refers to a non-aromatic monocyclic or polycyclic ring (fused, spiro or bridged) system having 1 to 8 heteroatoms selected from O, S or N, and optionally are benzofused or fused heteroaryl having 5-6 ring members and/or optionally are substituted, wherein the substituents are selected from halogen (e.g., F, Cl, Br, I), hydroxy, alkyl, alkenyl, alkynyl, cycloalkyl, acyl, acylamino, optionally substituted thiocarbonyl, optionally substituted aryl, alkoxy, alkoxyamino, alkaryl, cyano, nitro, oxo, -CηO, -OCF ₃ , -CF ₃ , -SCF ₃ , carboxy, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, -O-C(=O)R _λ , -O-C(=O)OR _λ , -C(=0)NR _λ R _π , SO _m R _ψ , -0-C(=0)NR _λ R _π , -NR _λ C(=0)0R _λ , -NR _λ C(=0)NR _λ R _π , -NR _λ R _π , mercapto, haloalkyl, thioalkyl, -COOR _ψ , -C00NηR _λ , -COR _λ ,

(wherein m, R _ψ , Rx and R _π are as defined earlier) or guanidine. Heterocyclyl can optionally include rings having one or more double bonds. Such ring systems can be mono-, bi- or tricyclic. Carbonyl or sulfonyl group can replace carbon atom(s) of heterocyclyl. Unless otherwise constrained by the definition, the substituents are attached to the ring atom, i.e., carbon or heteroatom in the ring. Also, unless otherwise constrained by the definition, the heterocyclyl ring optionally may contain one or more olefinic bond(s). Examples of heterocyclyl groups includes but are not limited to are tetrahydrofuranyl, dihydrofuranyl, dihydropyridinyl, dihydrobenzofuryl, azabicyclohexyl, dihydroindolyl, piperidinyl, isoxazolinyl, thiazolinyl, thiazolidinonyl, oxazolinyl, oxazolidinonyl, azabicyclo[3.1.0]hexyl, diazabicyclo[2.2.1]heptyl, azabicyclooctyl, azetidinyl, 1,4-benzodioxanyl, 1,3-benzodioxolyl, dihydrobenzofuryl, dihydroimidazolyl, dihydropyranyl, dihydrofuranyl, dihydroindolyl, dihydroisoxazolyl, dihydropyridinyl, dioxanyl, dioxolanyl, homopiperi- dinyl, imidazolinyl, imidazolidinyl, imidazopyridinyl, indolinyl, indolyl, isoindolel,3-dione, isothiazolidinyl, morpholinyl, napthyridinyl, oxazolidinyl, oxazolyl, phenoxazinyl, phenothiazinyl, piperazinyl, purinyl, pyrazinyl, pyrazolinyl, pyrazolyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, pyrrolyl, pyrrolopyridinyl, tetrahydropyranyl, tetrazolyl, thiazolidinyl and thiazolyl, and thienyl and the like.

"Heteroarylalkyl" refers to alkyl-heteroaryl group linked through alkyl portion, wherein the alkyl and heteroaryl are as defined earlier. "Heterocyclylalkyl" refers to alkyl-heterocyclyl group linked through alkyl portion, wherein the alkyl and heterocyclyl are as defined earlier.

"Acyl" refers to -C(=O)R ₀₀ wherein R ₀₀ is selected from hydrogen, alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, heterocyclyl, heteroarylalkyl or heterocyclylalkyl.

"Alkylcarbonyl" refers to -C(=O)R ₀₀ , wherein R ₀₀ is selected from alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, heterocyclyl, heteroarylalkyl or heterocyclylalkyl.

"Alkylcarboxy" refers to -O-C(=O)R _00> wherein R ₀₀ is selected from alkyl, cycloalkyl, aryl, aralkyl, heteroaryl, heterocyclyl, heteroarylalkyl or heterocyclylalkyl.

The term "polymorphs" refers to all crystalline forms and amorphous forms of the compounds described herein. In addition, some of the compounds described herein may

form solvates with water or common organic solvents. Such solvates are also encompassed within the scope of this invention.

The phrase "pharmaceutically acceptable carriers" is intended to include non- toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation auxiliary of any type.

The compounds of present invention include stereoisomers. The term "stereoisomer" refers to compounds, which have identical chemical composition, but differ with regard to arrangement of the atoms and the groups in space. These include enantiomers, diastereomers, geometrical isomers, atropisomers and comformational isomers. All such isomeric forms of these compounds are expressly included in the present invention. Each stereogenic carbon may be of the R or S configuration. Although specific compounds may be depicted in a particular stereochemical configuration, compounds having either the opposite stereochemistry at any given chiral center or mixtures thereof are envisioned as part of the invention. Geometric isomers may occur when a compound contains a double bond or some other feature that gives the molecule a certain amount of structural rigidity. An enantiomer is a stereoisomer of a reference molecule that is the nonsuperimposable mirror image of the reference molecule. A diastereomer is a stereoisomer of a reference molecule that has a shape that is not the mirror image of the reference molecule. An atropisomer is a conformation of a reference compound that converts to the reference compound only slowly on the NMR or laboratory time scale. Conformational isomers (or conformers or rotational isomers or rotamers) are stereoisomers produced by rotation about σ bonds, and are often rapidly interconverting at -25 ⁰ C. Racemic mixtures are also encompassed within the scope of this invention.

The compounds described herein exhibit significant potency in terms of their activity, as determined by in vitro receptor binding and functional assays and in vivo experiments. The compounds that were found active in vitro were tested in vivo. Some of the compounds are potent muscarinic receptor antagonists with high affinity towards M ₁ and M ₃ receptors. Therefore, pharmaceutical compositions for the possible treatment of the disease or disorders associated with muscarinic receptors are provided. Compounds disclosed herein may be prepared, for example, by techniques well known in the organic synthesis and familiar to a practitioner ordinarily skilled in art of this

invention. In addition, the processes described herein may enable the synthesis of the compounds of the present invention. However, these may not be the only means by which the compounds described in the invention may be synthesized. Further, the various synthetic steps described herein may be performed in alternate sequences in order to furnish the desired compounds.

Scheme I

Formula I

Formula IV N-alkylation R ₂ -Z Formula IVa

The compounds of Formula V can be prepared following the procedure as described in Scheme I. The compound of Formula II (wherein Gi is hydroxyl or -O-alkyl; R ₁ is the same as defined earlier) can be coupled with a compound of Formula III (wherein Y is -OH, -O-mesyl, -O-tosyl or -O-triflyl) to give a compound of Formula IV, which can be further quaternized with a compound of Formula IVa (wherein Z and R ₂ are the same as defined earlier) to give a compound of Formula V.

The coupling of a compound of Formula II with a compound of Formula III (when Gi is -O-alkyl and Y is -OH) can be carried out in an organic solvent, for example, toluene, hexane, heptane or xylene in the presence of sodium metal.

Alternatively, the coupling of a compound of Formula II with a compound of Formula III (when Gi is -O-alkyl; Y is -OH) can be carried out in an organic solvent, for example, tetrahydrofuran, dimethylformamide, diethyl ether or dioxane in the presence of a coupling agent, for example, carbonyldiimidazole, l-(3-dimethylaminopropyl)-3- ethylcarbodiimide hydrochloride (EDCHCl) or dicyclohexylcarbodiimide (DCC).

Also, coupling of a compound of Formula II with a compound of Formula III (when G ₁ is -O-alkyl; Y is -OH) can be carried out in an organic solvent for example, toluene, heptane or xylene in the presence of a base for example, sodium hydride or sodium alkoxide to give a compound of Formula IV. The coupling of a compound of Formula II with a compound of Formula III (when

Gi is -OH; Y is -0-mesyl, -O-tosyl or -O-triflyl) can be carried out in an organic solvent for example, toluene, heptane or xylene in the presence of a base for example, 1, 8- diazabicyclo[5.4.0]undecen-7-ene (DBU) or l,4-diazabicyclo[2.2.2]octane to give a compound of Formula IV. The quaternization of a compound of Formula IV to give a compound of Formula

V can be carried out by reacting the compound of Formula IV with a compound of Formula IVa in the presence of an organic solvents selected from acetonitrile, dichloromethane, dichloroethane, carbon tetrachloride, chloroform, toluene, benzene, dimethyl formamide dimethylsulphoxide or mixtures thereof. The following compounds are prepared by Scheme I:

9-(4-Fluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-meth yl-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 1);

7-[2-Hydroxy(di-2-thienyl)acetoxy]-9-methyl-9-(4-methylbenzy l)-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 2); 9-(4-Bromobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-methy l-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 3);

9-Benzyl-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-methyl-3-oxa-9 - azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 4);

9-(2,5-Difluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9- methyl-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 5);

9-(3-Bromobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-methy l-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 6);

9-(l,3-Benzodioxol-5-ylmethyl)-7-[2-hydroxy(di-2-thienyl)ace toxy]-9-methyl-3- oxa-9-azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 7);

9-(3,5-Difluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9- methyl-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 8); 9-(3-Fluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-meth yl-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 9);

9-(2-Fluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-meth yl-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 10);

9-(3,4-Difluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9- methyl-3-oxa-9- azonia-tricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 11) or

9-(2,5-Dibromobenzyl)-7-[2- hydroxy(di-2-thienyl)acetoxy]-9-methyl-3-oxa-9- azonia-tricyclo[3.3.1.0 ² ' ⁴ ] nonane bromide (Compound No. 12).

In the above scheme, where specific bases, condensing agents, protecting groups, catalysts, temperatures, etc. are mentioned, it is to be understood that other bases, condensing agents, protecting groups, solvents, catalysts, temperatures, etc. known to those skilled in the art may be used. Similarly, the reaction temperature and duration may be adjusted according to the desired needs.

The compounds described herein can be produced and formulated as their enantiomers, diastereomers, N-Oxides, polymorphs and pharmaceutically acceptable solvates, as well as their metabolites. Pharmaceutical compositions comprising the molecules of Formula I or metabolites, enantiomers, diastereomers, N-oxides, polymorphs, or pharmaceutically acceptable solvates thereof, in combination with pharmaceutically acceptable carrier and optionally included excipient can also be produced.

Where desired, the compounds of Formula I and/or their pharmaceutically acceptable solvates, stereoisomers, tautomers, racemates, prodrugs, metabolites, polymorphs or N-oxides may be advantageously used in combination with one or more other therapeutic agents. Examples of other therapeutic agents, which may be used in combination with compounds of Formula I of this invention and/or their pharmaceutically acceptable salts, pharmaceutically acceptable solvates, stereoisomers, tautomers, racemates, prodrugs, metabolites, polymorphs or N-oxides include but are not limited to, histamine antagonist, corticosteroids, beta agonist, leukotriene antagonist, EGFR kinase inhibitors, PAF antagonist, 5-lipoxygenase inhibitors, chemokine inhibitors, PDE-4 inhibitors or p38 MAP kinase inhibitors.

The compositions can be administered by inhalation.

Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable, aqueous or organic solvents, or mixtures thereof, and powders. The liquid or solid compositions may contain suitable pharmaceutically acceptable excipients. The compositions can be administered by the nasal respiratory route for local or systemic effect. Compositions can be nebulized by use of inert gases. Nebulized solutions may be breathed directly from the nebulizing device or the nebulizing device can be attached to a face masks tent, or intermittent positive pressure breathing machine. Solution, suspension or powder compositions can be administered nasally from devices, which deliver the formulation in an appropriate manner.

Alternatively, compositions can be administered orally, rectally, parenterally (intravenously, intramuscularly or subcutaneously), intracisternally, intravaginally, intraperitoneally or topically.

Solid dosage forms for oral administration may be presented in discrete units, for example, capsules, cachets, lozenges, tablets, pills, powders, dragees or granules, each containing a predetermined amount of the active compound. In such solid dosage forms, the active compound is admixed with at least one inert customary excipient (or carrier) such as sodium citrate or dicalcium phosphate or (a) fillers or extenders, as for example, starches, lactose, sucrose, glucose, mannitol and silicic acid, (b) binders, as for example, carboxymethylcellulose, alignates, gelatin, polyvinylpyrrolidone, sucrose and acacia, (c) humectants, as for example, glycerol, (d) disintegrating agents, as for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates and sodium carbonate, (e) solution retarders, as for example paraffin, (f) absorption accelerators, as for example, quaternary ammonium compounds, (g) wetting agents, as for example, cetyl alcohol and glycerol monostearate, (h) adsorbents, as for example, kaolin and bentonite and (i) lubricants, as for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols, and the like.

Solid dosage forms can be prepared with coatings and shells, such as enteric coatings and others well known in this art. They may contain opacifying agents, and can also be of such composition that they release the active compound or compounds in a certain part of the intestinal tract in a delayed manner. Examples of embedding compositions which can be used are polymeric substances and waxes.

The active compounds can also be in micro-encapsulated form, if appropriate, with one or more of the above mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, as for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, in particular, cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan or mixtures of these substances, and the like.

Besides such inert diluents, the composition can also include adjuvants, for example, wetting agents, emulsifying and suspending agents, sweetening, flavoring and perfuming agents, colorants or dyes. Suspensions, in addition to the active compounds, may contain suspending agents, as for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminium metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances, and the like.

Dosage forms for topical administration of a compound of this invention include powder, spray, inhalant, ointment, creams, salve, jelly, lotion, paste, gel, aerosol or oil. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservatives, buffers or propellants as may be required. Opthalmic formulations, eye ointments, powders and solutions are also contemplated as being within the scope of this invention.

Compositions suitable for parenteral injection may comprise pharmaceutically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions and sterile powders for reconstitution into sterile injectable solutions or dispersions. These preparations may contain anti-oxidants, buffers, bacteriostats and solutes, which render the compositions isotonic with the blood of the intended recipient. Aqueous and non-aqueous sterile suspensions may include suspending agents and thickening agents. The compositions may be presented in unit-dose or multi-dose containers, for example sealed ampoules and vials, and may be stored in a freeze-dried or lyophilized condition requiring only the addition of the sterile liquid carrier, for example, saline or water-for-injection immediately prior to use. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, and the like), suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions and by the use of surfactants.

These compositions may also contain adjuvants such as preserving, wetting, emulsifying, and dispensing agents. Prevention of the action of microorganisms can be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agents, for example sugars, sodium chloride and the like. Prolonged absorption of the injectable pharmaceutical form can be brought about by the use of agents delaying absorption, for example, aluminum monosterate and gelatin.

Suppositories for rectal administration of the compound of Formula I can be prepared by mixing the drug with a suitable nonirritating excipient such as cocoa butter and polyethylene glycols or a suppository wax, which are solid at ordinary temperatures but liquid at body temperature and which therefore melt in the rectum or vaginal cavity and release the drug.

If desired, and for more effective distribution, the compounds can be incorporated into slow release or targeted delivery systems such as polymer matrices, liposomes, and microspheres. They may be sterilized, for example, by filtration through a bacteria- retaining filter, or by incorporating sterilizing agents in the form of sterile solid

compositions which can be dissolved in sterile water, or some other sterile injectable medium immediately before use.

Actual dosage levels of active ingredient in the compositions of the invention and spacing of individual dosages may be varied so as to obtain an amount of active ingredient that is effective to obtain a desired therapeutic response for a particular composition and method of administration. It will be understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the compound chosen, the body weight, general health, sex, diet, route of administration, the desired duration of treatment, rates of absorption and excretion, combination with other drugs and the severity of the particular disease being treated and is ultimately at the discretion of the physician.

The pharmaceutical compositions described herein can be produced and administered in dosage units, each unit containing a certain amount of at least one compound described herein and/or at least one physiologically acceptable addition salt thereof. The dosage may be varied over extremely wide limits, as the compounds are effective at low dosage levels and relatively free of toxicity. The compounds may be administered in the low micromolar concentration, which is therapeutically effective, and the dosage may be increased as desired up to the maximum dosage tolerated by the patient. While the present invention has been described in terms of its specific embodiments, certain modifications and equivalents will be apparent to those skilled in the art and are included within the scope of the present invention. The examples are provided to illustrate particular aspects of the disclosure and do not limit the scope of the present invention as defined by the claims. Examples

Synthesis of methyl hydroxy(dithiophen-2-yl)acetate

The title compound was prepared following the procedure described in /. Am. Chem. Soc, 73, (5): (1951), 2216.

Synthesis of 9-methyl-3-oxa-9-azatricyclof3.3.1.0 ^{2 4} ]nonan-7-ol

To the suspension of hydrochloride salt of 9-methyl-3-oxa-9- azatricyclo[3.3.1.0 ² ' ⁴ ]nonan-7-ol (500 mg, 0.026 mol) in ethylacetate (30 mL) was added saturated aqueous solution of sodium hydroxide (1OmL) under cold condition until salt dissolved completely in ethyl acetate. The ethyl acetate layer was separated, dried over sodium sulphate (Na ₂ SO ₄ ), filtered, and concentrated under reduced pressure to get desired compound.

Example 1: Synthesis of 9-(4-fluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-meth yl- 3-oxa-9-azoniatricyclo[ 3.3.1.0 ² ' ⁴ lnonane bromide (Compound No. 1 )

Step a: Synthesis oi 9-methyl-3-oxa-9-azoniatricyclo[3.3.1.(f' ⁴ ]non-7-yl hydroxyl(dithiophen-2-yl) acetate

To the solution of methyl 2,2-di-2-thienylpropionate ( 819.35 mg, 0.0032 mol) and 9-methyl-3-oxa-9-azatricyclo[3.3.1.0 ² ' ⁴ ]nonan-7-ol (155 mg, 0.0032 mol) in dry toluene was added sodium metal (26.63 mg, 0.0011 mol) portion wise. The reaction mixture was refluxed at 80-90 ⁰ C and at 200 mbar reduced pressure with downward distillation. The methanol was distilled off and reaction mixture was refluxed for 5 hours. After completion of reaction, the reaction mixture was cooled to room temperature and poured into dilute hydrochloride and wet ice solution. The aqueous layer was separated and basified with aqueous sodium carbonate. The aqueous layer was extracted with dichloromethane. The dichloromethane layer was washed with water and brine, dried over anhydrous sodium sulphate and concentrated under reduced pressure. The residue thus obtained was purified by column chromatography using methanol: dichloromethane: ammonia (2:97:1) to get the desired compound.

Yield = 250 mg.

Mass spectrum (m/z, +ve ion mode): 378.1 (M ⁺ ). Step b: Synthesis oϊ 9-(4-fluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-meth yl-3- oxa-9-azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide

To a solution of compound obtained from step a (30 mg, 0.0001 mol) in the acetonitrile: dichloromethane (0.66ml: 0.33ml) was added 4-fluoro benzyl bromide (15 mg, 0.000 lmol) at room temperature. Reaction mixture was kept on stand for two days. The solid was separated out and filtered. The solid thus obtained was triturated with

diethyl ether and organic layer was decanted off. The solid was dried under high vacuum to get pure compound.

Yield = 43 mg.

Mass spectrum (m/z, +ve ion mode): 486.27 (M ⁺ ). The following compounds were prepared employing procedures as provided in

Example 1 described above:

7-[2-Hydroxy(di-2-thienyl)acetoxy]-9-methyl-9-(4-methylbenzy l)-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 2) Mass spectrum (m/z, +ve ion mode): 482 (M ⁺ ); 9-(4-Bromobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-methy l-3-oxa-9- azoniatricyclo[3.3.1.0 ² ' ⁴ ]nonane bromide (Compound No. 3) Mass spectrum (m/z, +ve ion mode): 547 (M ⁺ );

9-Benzyl-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-methyl-3-ox a-9- azoniatricyclo[3.3.1.02,4]nonane bromide (Compound No. 4) Mass spectrum (m/z, +ve ion mode): 468.24 (M+);

9-(2,5-Difluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy] -9-methyl-3-oxa-9- azoniatricyclo[3.3.1.02,4]nonane bromide (Compound No. 5) Mass spectrum (m/z, +ve ion mode): 503.92 (M+);

9-(3-Bromobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-me thyl-3-oxa-9- azoniatricyclo[3.3.1.02,4]nonane bromide (Compound No. 6)

Mass spectrum (m/z, +ve ion mode): 547.87 (M+);

9-(l,3-Benzodioxol-5-ylmethyl)-7-[2-hydroxy(di-2-thienyl) acetoxy]-9-methyl-3- oxa-9-azoniatricyclo[3.3.1.02,4]nonane bromide (Compound No. 7) Mass spectrum (m/z, +ve ion mode): 511.97 (M+); 9-(3,5-Difluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9- methyl-3-oxa-9- azoniatricyclo[3.3.1.02,4]nonane bromide (Compound No. 8) Mass spectrum (m/z, +ve ion mode): 503.9 (M+);

9-(3-Fluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-m ethyl-3-oxa-9- azoniatricyclo[3.3.1.02,4]nonane bromide (Compound No. 9) Mass spectrum (m/z, +ve ion mode): 485.98 (M+);

9-(2-Fluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy]-9-m ethyl-3-oxa-9- azoniatricyclo[3.3.1.02,4]nonane bromide (Compound No. 10) Mass spectrum (m/z, +ve ion mode): 485.95 (M+);

9-(3,4-Difluorobenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy] -9-methyl-3-oxa-9- azonia-tricyclo[3.3.1.02,4]nonane bromide (Compound No. 11)

Mass spectrum (m/z, +ve ion mode): 503.91(M+);

9-(2,5-Dibrormbenzyl)-7-[2-hydroxy(di-2-thienyl)acetoxy] -9-methyl-3-oxa-9- azonia-tricyclo[3.3.1.02,4] nonane bromide (Compound No. 12) Mass spectrum (m/z, +ve ion mode): 625.98 (M+).

Biological Activity

Radioligand Binding Assays:

The affinity of test compounds for Mi, M ₂ and M ₃ muscarinic receptor subtypes was determined by [ ³ H]-N-methylscopolamine binding studies using rat heart and submandibular gland respectively as described by Moriya et al., (Life ScL, 64 _. (25):

(1999), 2351 with minor modifications. In competition binding studies, specific binding of [ ³ H] νMS was also determined using membranes from Chinese hamster ovary (CHO) cells expressing cloned human Mi, M ₂ , M ₃ , M ₄ and M ₅ receptors. Selectivities were calculated from the K ₁ values obtained on these human cloned membranes. Membrane preparation: Submandibular glands and heart were isolated and placed in ice- cold homogenizing buffer (HEPES 2OmM, 1OmM EDTA, pH 7.4) immediately after sacrifice. The tissues were homogenized in 10 volumes of homogenizing buffer and the homogenate was filtered through two layers of wet gauze and filtrate was centrifuged at 500g for 10 minutes at 4 ⁰ C. The supernatant was subsequently centrifuged at 40,00Og for 20 minutes at 4 ⁰ C. The pellet thus obtained was resuspended in assay buffer (HEPES 20 niM, EDTA 5mM, pH 7.4) and were stored at -70 ⁰ C until the time of assay.

Ligand binding assay: The compounds were dissolved and diluted in DMSO. The membrane homogenates (150-250 μg protein) were incubated in 250 μl of assay volume

(HEPES 20 niM, pH 7.4) at 24-25 °C for 3 hours. Non-specific binding was determined in the presence of 1 μM atropine. The incubation was terminated by vacuum filtration over GF/B fiber filters (Wallac). The filters were then washed with ice-cold 5OmM Tris HCl buffer (pH 7.4). The filter mats were dried and bound radioactivity retained on filters was counted. The IC50 & K _d were estimated by using the non-linear curve fitting program using G Pad Prism software. The value of inhibition constant K ₁ was calculated from competitive binding studies by using Cheng & Prusoff equation (Biochem.

Pharmacol, 22, (1973), 3099-3109 K ₁ = IC ₅₀ /(1+17Kd), where L is the concentration of

[3H]NMS used in the particular experiment. pK, is -log [K ₁ ].

The tested compounds exhibited K ₁ at m3 receptor in the range of 0.5 nM to 30 nM.

Functional Experiments using isolated rat bladder: Methodology

Animals were euthanized by overdose of thiopentone and whole bladder was isolated and removed rapidly and placed in ice cold Tyrode buffer with the following composition (mMol/L) NaCl 137; KCl 2.7; CaCl ₂ 1.8; MgCl ₂ 0.1; NaHCO ₃ 11.9; NaH ₂ PO ₄ 0.4; Glucose 5.55 and continuously gassed with 95% O ₂ and 5 % CO ₂ .

The bladder was cut into longitudinal strips (3mm wide and 5-6 mm long) and mounted in 10 ml organ baths at 30 ⁰ C, with one end connected to the base of the tissue holder and the other end connected through a force displacement transducer. Each tissue was maintained at a constant basal tension of 1 g and allowed to equilibrate for 11/2 hours during which the Tyrode buffer was changed every 15-20 minutes. At the end of equilibration period the stabilization of the tissue contractile response was assessed with lμmol/L of carbachol till a reproducible response was obtained. Subsequently a cumulative concentration response curve to carbachol (10 ^~9 mol/L to 3 X 10 ^"4 mol/L) was obtained. After several washes, once the baseline was achieved, cumulative concentration response curve was obtained in presence of NCE (NCE added 20 minutes prior to the second cumulative response curve). The contractile results were expressed as % of control E max. ED ₅₀ values are calculated by fitting a non-linear regression curve (Graph Pad Prism). pKb values were calculated by the formula pKb = - log [ (molar concentration of antagonist/ (dose ratio-1))] where, dose ratio = ED50 in the presence of antagonist/ED50 in the absence of antagonist.

In-vitro functional assay

Animals and anaesthesia: The Guinea Pig (400-600gm) was procured and trachea was removed under anesthesia (sodium pentobarbital, 300 mg/kg i.p) and was immediately kept in an ice-cold Krebs Henseleit buffer. Indomethacin (lOμM) was present throughout the KH buffer to prevent the formation of bronchoactive prostanoids.

Trachea experiments:

The tissue of adherent fascia was cleaned and was cut into strips of equal size (with approximately 4-5 tracheal rings in each strip). The epithelium was removed by careful rubbing, minimizing damage to the smooth muscle. Trachea was opened along the mid- dorsal surface with the smooth muscle band intact and made a series of transverse cuts from alternate sides so that they do not transect the preparation completely. Opposite ends of the cut rings were tied with the help of a thread. The tissue was mounted in isolated tissue baths containing 10ml Krebs Henseleit buffer maintained at 37 ⁰ C and bubbled with carbogen, at a basal tension of 1 gm. The buffer was changed 4-5 times for about an hour. The tissue was equilibrated for 1 hour for stabilization. After 1 hour, the tissue was challenged with lμM carbachol. This was repeated after every 2-3 washes till two similar consecutive responses were obtained. At the end of stabilization, the tissue was washed for 30 minutes followed by incubation with suboptimal dose of MRA/Vehicle for 20 minutes prior to contraction of the tissues with lμM carbachol. The contractile response of tissues was recorded either on Powerlab data acquisition system or on Grass polygraph (Model 7). The relaxation was expressed as percentage of maximum carbachol response. The data was expressed as mean ± s.e. mean for n observations. The EC ₅₀ was calculated as the concentration producing 50% of the maximum relaxation to lμM carbachol. The percent relaxation was compared between the treated and control tissues using non- parametric unpaired t-test. A p value of < 0.05 was considered to be statistically significant.

The tested compounds exhibited a pKb in the range of 8.2 to 9.2.

In-vitro functional assay to evaluate efficacy of "MRA" in combination with "PDE-IV inhibitors" Animals and anaesthesia:

Trachea tissue is obtained from a guinea pig (400-600gm) under anesthesia (sodium pentobarbital, 300 mg/kg i.p) and is immediately kept in an ice-cold Krebs Henseleit buffer. Indomethacin (lOμM) is present throughout the KH buffer to prevent the formation of bronchoactive prostanoids.

Trachea experiments:

Trachea tissue is cleaned off adherent fascia and cut it into strips of equal size (with approximately 4-5 tracheal rings in each strip). The epithelium is removed by careful rubbing, minimizing damage to the smooth muscle. The trachea is opened along the mid-dorsal surface with the smooth muscle band intact and a series of transverse cuts is made from alternate sides so that they do not transect the preparation completely. Opposite ends of the cut rings are tied with the help of a thread. The tissue is mounted in isolated tissue baths containing 10 mL Krebs Henseleit buffer maintained at 37 ⁰ C and is bubbled with carbogen, at a basal tension of 1 gm. The buffer is changed 4-5 times for about an hour and the tissue is equilibrated for 1 hour for stabilization. After 1 hour, the tissue is contacted with lμM carbachol. Repeat this after every 2-3 washes until two similar consecutive responses are obtained. At the end of stabilization, the tissue is washed for 30 minutes followed by incubation with suboptimal dose of MRA/Vehicle for 20 minutes prior to contraction of the tissues with lμM carbachol. The relaxant activity of the PDE-IV inhibitor [10 ^{^9} M to 10 ^{^4} M ] on the stabilized developed tension/response is assessed. The contractile response of tissues is recorded either on a Powerlab data acquisition system or on a Grass polygraph (Model 7). The relaxation is expressed as a percentage of maximum carbachol response. The data is expressed as mean ± s.e. mean for n observations. The EC ₅₀ is calculated as the concentration producing 50% of the maximum relaxation to lμM carbachol. The percent relaxation between the treated and control tissues is compared using non-parametric unpaired t-test. A p value of < 0.05 is considered to be statistically significant.

In-vivo assay to evaluate efficacy of MRA inhibitors

Male Guinea pigs were anesthetized with urethane (1.5 g/kg, Lp.). Trachea was cannulated along with jugular vein (for carbachol challenge) and animals were placed in the Plethysmograph-Box (PLY 3114 model; Buxco Electronics, Sharon, USA.). Respiratory parameters were recorded using Pulmonary Mechanics Analyzer, Biosystems XA software (Buxco Electronics, USA), which calculated lung resistance (R _L ) on a breath- by-breath basis. Bronchoconstriction was induced by injections of carbachol (10 μg/kg) delivered into the jugular vein. Increase in R _L over a period of 5 minutes post carbachol

challenge was recorded in presence or absence of MRA or vehicle at 2 hours and 12 hours post treatment and expressed as % increase in R _L from basal.

In-vivo assay to evaluate efficacy of MRA in combination with PDE-IV inhibitors

Drug treatment: MRA (lμg/kg to lmg/kg) and PDE-IV inhibitor (lμg/kg to lmg/kg) are instilled intratracheally under anesthesia either alone or in combination.

Method:

Male wistar rats weighing 200±20gm are used in the study. Rats have free access to food and water. On the day of experiment, animals are exposed to lipopoly saccharide (LPS, lOOμg/ml) for 40 minutes. One group of vehicle treated rats is exposed to phosphate buffered saline (PBS) for 40 minutes. Two hours after LPS/PBS exposure, animals are placed inside a whole body plethysmograph (Buxco Electronics, USA) and exposed to PBS or increasing acetylcholine (1, 6, 12, 24, 48 and 96 mg/ml) aerosol until Penh values (index of airway resistance) of rats attained 2 times the value (PC-100) seen with PBS alone. The respiratory parameters are recorded online using Biosystem XA software, (Buxco Electronics, USA). Penh, at any chosen dose of acetylcholine is, expressed as percent of PBS response and the using a nonlinear regression analysis PClOO (2 folds of PBS value) values are computed. Percent inhibition is computed using the following formula. PCIOOLPS - PCIOOTEST

% Inhibition = X 100

PCIOOLPS - PC100 _PBS Where,

PC 1 OO _LPS = PC 100 in untreated LPS challenged group PCIOO _TEST = PClOO in group treated with a given dose of test compound

PClOOpBs = PClOO in group challenged with PBS

Immediately after the airway hyper reactivity response is recorded, animals are sacrificed and bronchoalveolar lavage (BAL) is performed. Collected lavage fluid is centrifuged at 3000 rpm for 5 minutes at 4 ⁰ C. Total leukocyte count is performed in the

resuspended sample. A portion of suspension is cytocentrifuged and stained with Leishmann's stain for differential leukocyte count. Total leukocyte and neutrophil counts are expressed as cell count (millions cells ml ^"1 of BAL). Percent inhibition is computed using the following formula. NCLPS - NCTEST

% Inhibition = X 100

NCLPS - NCCON Where,

NC _LPS = Percentage of neutrophil in untreated LPS challenged group NC _TEST = Percentage of neutrophil in group treated with a given dose of test compound

NC _CON = Percentage of neutrophil in group not challenged with LPS

The percent inhibition data is used to compute ED ₅₀ vales using Graph Pad Prism software (Graphpad Software Inc., USA). In-vivo assay to evaluate efficacy of MRA in combination with Corticosteroids

Ovalbumin induced airway inflammation:

Guinea pigs are sensitized on days 0, 7 and 14 with 50-μg ovalbumin and 10 mg aluminum hydroxide injected intraperitoneally. On days 19 and 20 guinea pigs are exposed to 0.1% w v ^"1 ovalbumin or PBS for 10 minutes, and with 1% ovalbumin for 30 minutes on day 21. Guinea pigs are treated with test compound (0.1, 0.3 and 1 mg kg ^"1 ) or standard 1 mg kg ^"1 or vehicle once daily from day 19 and continued for 4 days. Ovalbumin/PBS challenge is performed 2 hours after different drug treatment.

Twenty four hours after the final ovalbumin challenge BAL is performed using Hank's balanced salt solution (HBSS). Collected lavage fluid is centrifuged at 3000 rpm for 5 minutes at 4 ⁰ C. Pellet is collected and resuspended in ImI HBSS. Total leukocyte count is performed in the resuspended sample. A portion of suspension is cytocentrifuged and stained with Leishmann's stain for differential leukocyte count. Total leukocyte and eosinophil count are expressed as cell count (millions cells ml ^"1 of BAL). Eosinophil is

also expressed as percent of total leukocyte count. % inhibition is computed using the following formula.

EOSOVA - EOSTEST % Inhibition = X 100 EOSOVA - EoscoN

Where,

EOS _OVA = Percentage of eosinophil in untreated ovalbumin challenged group

EOS _TEST = Percentage of eosinophil in group treated with a given dose of test compound EoscoN = Percentage of eosinophil in group not challenged with ovalbumin.

In-vivo assay to evaluate efficacy of "MRA" in combination with p38 MAP Kinase inhibitors

Lipopoly saccharide (LPS) induced airway hyper reactivity (AHR) and neutrophilia: Drug treatment:

MRA (lμg/kg to lmg/kg) and p38 MAP kinase inhibitor (lμg/kg to lmg/kg) are instilled intratracheally under anesthesia either alone or in combination.

Method:

Male wistar rats weighing 200±20gm are used in the study. Rats have free access to food and water. On the day of experiment, animals are exposed to lipopoly saccharide (LPS, lOOμg/ml) for 40 minutes. One group of vehicle treated rats is exposed to phosphate buffered saline (PBS) for 40 minutes. Two hours after LPS/PBS exposure, animals are placed inside a whole body plethysmograph (Buxco Electronics, USA) and exposed to PBS or increasing acetylcholine (1, 6, 12, 24, 48 and 96 mg/ml) aerosol until Penh values (index of airway resistance) of rats attained 2 times the value (PC-100) seen with PBS alone. The respiratory parameters are recorded online using Biosystem XA software, (Buxco Electronics, USA). Penh, at any chosen dose of acetylcholine is, expressed as percent of PBS response and the using a nonlinear regression analysis PClOO

(2 folds of PBS value) values are computed. Percent inhibition is computed using the following formula.

PCIOOLPS - PCIOOTEST % Inhibition = X 100 PCIOOLPS - PClOOpBs

Where,

PC 1 OO _LPS = PC 100 in untreated LPS challenged group

PCIOO _TEST = PClOO in group treated with a given dose of test compound

PClOOpBs = PClOO in group challenged with PBS Immediately after the airway hyper reactivity response is recorded, animals are sacrificed and bronchoalveolar lavage (BAL) is performed. Collected lavage fluid is centrifuged at 3000 rpm for 5 minutes at 4 ⁰ C. Pellet is collected and resuspended in ImI HBSS. Total leukocyte count is performed in the resuspended sample. A portion of suspension is cytocentrifuged and stained with Leishmann' s stain for differential leukocyte count. Total leukocyte and neutrophil counts are expressed as cell count

(millions cells ml ^"1 of BAL). Percent inhibition is computed using the following formula.

NCLPS — NCTEST % Inhibition = X 100

NCLPS - NC _CO N Where,

NC _LPS = Percentage of neutrophil in untreated LPS challenged group

NC _TEST = Percentage of neutrophil in group treated with a given dose of test compound

NCcoN = Percentage of neutrophil in group not challenged with LPS The percent inhibition data is used to compute ED ₅₀ values using Graph Pad Prism software (Graphpad Software Inc., USA).

In-vivo assay to evaluate efficacy of "MRA" in combination with β2-agonists

Drug treatment:

MRA (lμg/kg to lmg/kg) and long acting β ₂ agonist is instilled intratracheally under anesthesia either alone or in combination. Method:

Wistar rats (250-350gm) or balb/C mice (20-30gm) is placed in body box of a whole body plethysmograph (Buxco Electronics, USA) to induce bronchoconstriction. Animals are allowed to acclimatize in the body box and are given successive challenges, each of 2 minute duration, with PBS (vehicle for acetylcholine) or acetylcholine (i.e. 24, 48, 96, 144, 384, and 768 mg/ml). The respiratory parameters are recorded online using Biosystem XA software, (Buxco Electronics, USA) for 3 minutes. A gap of 2 minutes is allowed for the animals to recover and then challenged with the next higher dose of acetylcholine (ACh). This step is repeated until Penh of rats attained 2 times the value (PC-100) seen with PBS challenge. Following PBS/ACh challenge, Penh values (index of airway resistance) in each rat/mice is obtained in the presence of PBS and different doses of ACh. Penh, at any chosen dose of ACh is, expressed as percent of PBS response. The Penh values thus calculated are fed into Graph Pad Prism software (Graphpad Software Inc., USA) and using a nonlinear regression analysis PClOO (2 folds of PBS value) values are computed. % inhibition is computed using the following formula. PCIOOTEST - PCIOOCON

% Inhibition = X 100

768 - PClOOcoN Where,

PClOOcoN = PClOO in vehicle treated group PCIOO _TEST = PClOO in group treated with a given dose of test compound

768 = is the maximum amount of acetylcholine used.