GLYCINE DERIVATIVES AND THEIR USE AS MUSCARINIC RECEPTOR ANTAGONISTS

FIELD OF THE INVENTION

The present invention relates to glycine derivatives acting as muscarinic receptor antagonists, processes for their preparation, compositions comprising them and therapeutic uses thereof.

BACKGROUND OF THE INVENTION

Quaternary ammonium salts acting as muscarinic (M) receptor antagonist drugs are currently used in therapy to induce bronchodilation for the treatment of respiratory diseases. Examples of well known M receptor antagonists are for instance represented by ipratropium bromide and tiotropium bromide.

Several chemical classes acting as selective M3 receptor antagonist drugs have been developed for the treatment of inflammatory or obstructive airway diseases such as asthma and chronic obstructive pulmonary disease (COPD).

Quinuclidine carbamate derivatives and their use as M3 antagonists are disclosed e.g. in WO 02/051841, WO 03/053966 and WO 2008/012290.

Said M and M3 receptor antagonists are currently administered through inhalation route in order to deliver the drug directly at the site of action, thus limiting the systemic exposure and any undesirable side effect due to systemic absorption. However, even though the systemic exposure may be reduced through the inhalatory route, the compounds of the prior art may still, at least potentially, exhibit undesired side effects due to systemic absorption.

Therefore, it is highly desirable to provide M3 receptor antagonists able to act locally, while having high selectivity and plasmatic instability. Said drugs, once adsorbed, are degraded to inactive compounds which are deprived of any systemic side effects typical of muscarinic antagonists.

WO 2010/072338, describes azonia-bicyclo[2.2.2]octane compounds acting as muscarinic receptor antagonists, further possessing the above therapeutically desirable characteristics.

Surprisingly, it has been now found that the glycine derivatives, as per the details below, improve even further the selectivity and plasmatic instability with respect to these latter compounds, with the consequent minimization of side effects.

The compounds of the present invention thus behave as soft-drugs, since they are able to produce a more persistent bronchodilating effect in the lungs but are more consistently and rapidly transformed into inactive metabolites after passing into human plasma.

This behaviour gives great advantages in terms of safety.

SUMMARY OF THE INVENTION

The present invention thus relates to glycine derivatives of general formula (I) or (VI), acting as muscarinic receptor antagonists, processes for their preparation, compositions comprising them, therapeutic uses and combinations with other pharmaceutical active ingredients among which are, for instance, those currently used in the treatment of respiratory disorders, e.g. beta2-agonists, corticosteroids, P38 MAP kinase inhibitors, IKK2 inhibitors, HNE inhibitors, PDE4 inhibitors, leukotriene modulators, NSAIDs and mucus regulators.

DETAILED DESCRIPTION OF THE INVENTION

In particular, the invention is directed to glycine derivatives of general formula (I)

(I) wherein:

Rl is selected from the group consisting of -H, linear or branched (Ci-Cio)alkyl, (C ₂ -C ₆ )alkenyl, aryl, (C ₃ -C ₈ )cycloalkyl,

(C ₅ -C ₁₀ )heterocycloalkyl, aryl(C ₁ -C ₆ )alkyl and heteroaryl, optionally substituted by one or more substituents selected from the group consisting of halogen atoms, -OH, oxo (=O), -SH, -NO ₂ , -N(R5)(R6), -CN, -CON(R5) ₂ , -NHCO(R5), -COR5, -CO ₂ R5, (C ₁ -C ₁₀ )alkylsulfanyl, (C ₁ -C ₁₀ )alkylsulfinyl, (C ₁ -C ₁₀ )alkylsulfonyl, (C ₁ -C ₁₀ )alkyl, (C ₁ -C ₁₀ )alkylcarboxyl, (C ₁ -C ₁₀ )alkoxy, aryl, aryloxy and heteroaryl;

G is selected from the group consisting of -OC(O)-, -SO ₂ - and -C(O)-;

R2 is selected from the group consisting of -H, (C ₁ -C ₁₀ )alkyl and aryl(C ₁ -C ₆ )alkyl, optionally substituted by one or more substituents selected from the group consisting of halogen atoms, -OH, oxo (=O), -SH, -NO ₂ , -N(R5) ₂ , -CN, -CON(R5) ₂ , -NHCO(R5), -CO(R5), -CO ₂ (R5), (C ₁ -C ₁₀ )alkylsulfanyl, (C ₁ -C ₁₀ )alkylsulfinyl, (C ₁ -C ₁₀ )alkylsulfonyl,

(C ₁ -C ₁₀ )alkyl, (C ₁ -C ₁₀ )alkylcarboxyl, (C ₁ -C ₁₀ )alkoxy, aryl, aryloxy and heteroaryl;

R3 is selected from the group consisting of -H, (C ₁ -C ₁₀ )alkyl, aryl, (C ₃ -C ₈ )cycloalkyl, heteroaryl, aryl(C ₁ -C ₆ )alkyl and heteroaryl(C ₁ -C ₆ )alkyl, optionally substituted by one or more substituents selected from the group consisting of halogen atoms, -OH, oxo (=O), -SH, -NO ₂ , -N(R5) ₂ , -CN, -CON(R5) ₂ , -NHCO(R5), -CO(R5), -CO ₂ (R5), (C ₁ -C ₁₀ )alkylsulfanyl, (C ₁ -C ₁₀ )alkylsulfinyl, (C ₁ -C ₁₀ )alkylsulfonyl, (C ₁ -C ₁₀ )alkyl,

(C ₁ -C ₁₀ )alkylcarboxyl, (C ₁ -C ₁₀ )alkoxy, aryl, aryloxy, aryl(C ₁ -C ₁₀ )alkylenoxy and heteroaryl;

R6 is selected from the group consisting of residues of formula (i), (ii), (iii) and (iv)

(i) (ϋ)

(iii) (iv)

wherein

m = 1, 2 or 3 ;

n = 1, 2 or 3 ;

A ^" is a physiologically acceptable anion;

R4 is a group of formula (Y)

— (CH ₂ )p— P— (CH ₂ )q— W

(Y)

wherein

p is 0 or an integer from 1 to 4;

q is 0 or an integer from 1 to 4;

P is absent or is selected from the group consisting of -O-, -S-, -SO-, -SO ₂ -, -C(O)-, -N(R5)- -CH=CH-, -N(R5)(SO ₂ )-, -N(R5)(COO)-, -N(R5)(C(O))-, -S(O ₂ )N(R5)-, -CO(O)N(R5)- and -C(O)N(R5)-;

W is selected from the group consisting of H, linear or branched (C ₁ -C ₁₀ )alkyl, (C ₂ -C ₆ )alkenyl, aryl, (C ₃ -C ₈ )cycloalkyl,

(C ₅ -C ₁₀ )heterocycloalkyl, aryl(C ₁ -C ₆ )alkyl and heteroaryl, optionally substituted by one or more substituents selected from the group consisting of halogen atoms, -OH, oxo (=O), -SH, -NO ₂ , -N(R5) ₂ , -CN, -CON(R5) ₂ , -NH(COR5), -CO(R5), -CO ₂ (R5), (C ₁ -C ₁₀ )alkylsulfanyl, (C ₁ -C ₁₀ )alkylsulfinyl, (C ₁ -C ₁₀ )alkylsulfonyl, (C ₁ -C ₁₀ )alkyl, (C ₁ -C ₁₀ )alkylcarboxyl, (C ₁ -C ₁₀ )alkoxy, aryl, aryloxy and heteroaryl;

R5 and R6 are independently selected from the group consisting of -H, (C ₁ -C ₁₀ )alkyl, (C ₁ -C ₁₀ )alkoxy, (C ₂ -C ₆ )alkynyl, (C ₂ -C ₆ )alkenyl, (C ₃ -C ₇ )cycloalkyl, (C ₃ -C ₇ )cycloalkyl-(C ₁ -C ₁₀ )alkyl, heteroaryl, (C ₁ -C ₁₀ )alkyl- heteroaryl and aryl optionally substituted by one or more substituents selected from the group consisting of halogen atoms, -OH, oxo (=O), -SH, -NO ₂ , -CN, -CONH ₂ , (C ₁ -C ₁₀ )alkylsulfanyl, (C ₁ -C ₁₀ )alkylsulfinyl, (C ₁ -C ₁₀ )alkylsulfonyl, (C ₁ -C ₁₀ )alkyl, (C ₁ -C ₁₀ )alkylcarboxyl, (C ₁ -C ₁₀ )alkoxy, aryl, aryloxy and heteroaryl and

pharmaceutically acceptable salts thereof.

The present invention is also directed to compounds of general formula (VI):

(VI)

wherein:

Rl, R2, R3 and G are as defined above;

selected from the group consisting of residues of formula (v),

(vi), (vii) and (viii)

m and n are as defined above.

The asterisk within groups of formula (i) to (viii), denotes the point of attachment with the rest of the molecule.

In the present disclosure, unless otherwise specified, the term "halogen" includes fluorine, chlorine, bromine and iodine atoms.

The expression "(C ₁ -C ₁₀ )alkyl", refers to straight or branched chain alkyl groups wherein the number of carbon atoms is from 1 to 10. Examples of said groups are methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl and the like.

The expression "(C ₂ -C ₆ )alkenyl" refers to straight or branched carbon chains with one or more double bonds. Examples of said groups are ethenyl, propenyl, butenyl, pentenyl, hexenyl and the like.

The expression "(C ₁ -C ₁₀ )alkoxy" refers to the above alkyl-oxy (e.g. alkoxy) groups. Examples of said groups are methoxy, ethoxy, n-propoxy, isopropoxy, n- butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentoxy, hexoxy and the like.

Likewise, the expressions "(C ₁ -C ₁₀ )alkylsulfanyl",

"(Ci-Cio)alkylsulfinyl", "(C ₁ -C ₁₀ )alkylsulfonyl" or "(C ₁ -C ₁₀ )alkylcarboxyl refer, respectively, to alkyl-S-, alkyl-SO-, alkyl-SO ₂ - or alkyl-COO groups. The expression "(C ₃ -C ₈ )cycloalkyr' refers to cyclic non-aromatic hydrocarbon groups with 3 to 8 carbon atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.

The expression "(C ₅ -C ₁₀ )heterocycloalkyl" refers to (C ₅ -C ₁₀ )cycloalkyl wherein one or more hydrogen atoms are replaced by one or more halogen atoms, which can be the same or different.

The expression "aryl" refers to mono or bi- or tricyclic ring systems which have 6 to 20 ring atoms, preferably 6 to 15 and wherein at least one ring is aromatic.

The expressions "aryl(C ₁ -C ₆ )alkyl", "heteroaryl(C ₁ -C ₆ )alkyl and (C ₃ -C ₈ )cycloalkyl(C ₁ -C ₆ )alkyl refer to (C ₁ -C ₆ )alkyl groups further substituted by aryl, heteroaryl or cycloalkyl rings.

The expression "aryloxy" refers to -O-aryl group. An example may be phenyloxy.

The expression "(C ₁ -C ₁₀ )alkylene" refers to a chain with 1 to 10 -CH ₂ - groups. An example may be methylene.

The expression "(C ₁ -C ₁₀ )alkylenoxy" refers to -O(C ₁ -C ₁₀ )alkylene.

The expression "aryl(C ₁ -C ₁₀ )alkylenoxy" refers to (C ₁ -C ₁₀ )alkylenoxy further substituted by aryl. An example may be benzyloxy.

The expression "heteroaryl" refers to mono, bi- or tricyclic ring systems which have 5 to 20 ring atoms, preferably 5 to 15, in which at least one ring is aromatic and in which at least one ring atom is a heteroatom or hetero aromatic group (e.g. N, NH, S or O).

Examples of suitable aryl or heteroaryl monocyclic systems include for instance thiophene, benzene, pyrrole, pyrazole, imidazole, isoxazole, oxazole, oxadiazole, isothiazole, thiazole, pyridine, imidazolidine and furan residues and the like.

Examples of suitable aryl or heteroaryl bicyclic systems include benzodioxole, naphthalene, biphenylene, purine, pteridine, benzotriazole, quinoline, isoquinoline, indole, isoindole, benzothiophene, dihydrobenzo dioxin, dihydrobenzo dioxepin and benzo oxazine residues and the like.

Advantageously, the physiologically acceptable anions A ^" include those selected from chloride, bromide, iodide, trifluoroacetate, formate, sulfate, phosphate, methane sulfonate, nitrate, maleate, acetate, citrate, fumarate, tartrate, oxalate, succinate, benzoate and p-toluenesulfonate.

Besides the presence of A ^" anion, whenever further basic amino groups are present in the compounds of formula (I), additional physiological acceptable anions, among those formerly indicated, may be present. Likewise, in the presence of acidic groups such as COOH groups, corresponding physiological cation salts may be present as well, for instance including alkali or earth-alkali metal ions.

A first preferred group of compounds of general formula (I) or (VI) is that wherein G is selected from the group consisting of -OC(O)-, -SO ₂ - and -C(O)-, Rl is selected from the group consisting of linear or branched (Ci-Cio)alkyl, aryl, (C ₃ -C ₈ )cycloalkyl, aryl(C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₅ -C ₁₀ )heterocycloalkyl and heteroaryl, optionally substituted by one or more substituents selected from halogen atoms, -N(R5)(R6), (C ₁ -C ₁₀ )alkyl, (C ₁ -C ₁₀ )alkylcarboxyl, (C ₁ -C ₁₀ )alkoxy, aryloxy and heteroaryl; R2 is H; and R3 and R6 are as defined above.

Still more preferred, within this class, are the compounds of general formula (I) or (VI) wherein Rl is selected from the group consisting of methyl, ethyl, methoxyethoxyl, tert-butyl, ethenyl, cyclohexyl, phenyl, methoxyphenyl, chlorophenyl, difluorophenyl, dimethylthiazole, trifluoroethyl, phenylethyl, cyclopentyl, methylethoxyl, oxo-phenylethyl, thiophenyl, thiazolyl, fluorophenyl, amino-phenyl, tert-butoxycarbonylamino- phenyl and methylphenyl. Another preferred group of compounds of general formula (I) or (VI) within this class, is that wherein G, Rl and R2 are as defined above; R3 is selected from the group consisting of (C ₁ -C ₁₀ )alkyl, aryl and heteroaryl, optionally substituted by one or more groups selected from halogen atoms, (C ₁ -C ₁₀ )alkyl, (C ₁ -C ₁₀ )alkoxy and aryl(C ₁ -C ₁₀ )alkylenoxy; and R6 is as defined above.

An even more preferred group of compounds of general formula (I) or (VI) is that wherein G, Rl, R2 and R3 are as defined above; R6 is selected from the group consisting of residues of formula (i), (ii) and (iii), wherein A ^" is as defined above, R4 is a group of formula (Y) wherein p is 0, 1 and 3, P is -C(O)-, q is 0, W is selected from the group consisting of (C ₁ -C ₁₀ )alkyl, aryl, heteroaryl, (C ₅ -C ₁₀ )heterocycloalkyl, optionally substituted by one or more substituents selected from the group consisting of halogen atoms, (C ₁ -C ₁₀ )alkyl, (C ₁ -C ₁₀ )alkoxy, -OH and (C ₁ -C ₁₀ )alkylcarboxyl.

Still more preferred, within this class, are the compounds of general formula (I) or (VI) wherein W is selected from the group consisting of phenyl, benzothioxol, thiophenyl and thiazolyl, optionally substituted by one or more halogen atoms, OH, methyl and methylcarboxyl.

According to specific embodiments, the present invention provides the following compounds:

(continued) (3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l-(2-( 2-

C6

fluorophenyl)-2-oxoethyl)-l-azoniabicyclo[2.2.2]octane bromide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l-( 2-(2,4-

C7

difluorophenyl)-2-oxoethyl)- l-azoniabicyclo[2.2.2]octane bromide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l-( 2-(4-

C8

chlorophenyl)-2-oxoethyl)-l-azoniabicyclo[2.2.2]octane bromide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l-( 2-oxo-2-p-

C9

tolylethyl)-l-azoniabicyclo[2.2.2]octane bromide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l-(2-( 4-

CIO

methoxyphenyl)-2-oxoethyl)-l-azoniabicyclo[2.2.2]octane bromide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l-( 2-(4-

Cll

hydroxyphenyl)-2-oxoethyl)-l-azoniabicyclo[2.2.2]octane bromide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l-( 2-(4-

C12 (methoxycarbonyl)phenyl)-2-oxoethyl)-l-azoniabicyclo[2.2.2]o ctane bromide

(3R)- 1 -(2-(benzo[d] [ 1 ,3 ]dioxol-5-yl)-2-oxoethyl)-3-(2-(tert-

C13 butoxycarbonylamino)-2-phenylacetoxy)-l-azoniabicyclo[2.2.2] octane bromide

(3R)-l-(2-(benzo[b]thiophen-5-yl)-2-oxoethyl)-3-(2-(tert-

C14 butoxycarbonylamino)-2-phenylacetoxy)-l-azoniabicyclo[2.2.2] octane bromide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l-(2-( 5-

C15

chlorothiophen-2-yl)-2-oxoethyl)- l-azoniabicyclo[2.2.2]octane bromide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l -(2-0X0-2-

C16

(thiazol-2-yl)ethyl)-l-azoniabicyclo[2.2.2]octane bromide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l -(2-0X0-2-

C17

(thiophen-3-yl)ethyl)-l-azoniabicyclo[2.2.2]octane bromide

(3R)-3 -(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)- 1 -methyl- 1 -

C18

azoniabicyclo[2.2.2]octane iodide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l-(3-

C19

phenoxypropyl)-l-azoniabicyclo[2.2.2]octane bromide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l-(2-( 2-

C20

methoxyphenyl)-2-oxoethyl)-l-azoniabicyclo[2.2.2]octane bromide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l-( (5-phenyl-

C21

l,2,4-oxadiazol-3-yl)methyl)-l-azoniabicyclo[2.2.2]octane chloride

(3R)-l-(2-tert-butoxy-2-oxoethyl)-3-(2-(tert-butoxycarbon ylamino)-2-

C22

phenylacetoxy)-l-azoniabicyclo[2.2.2]octane bromide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l-(2-( 2-

C23

nitrophenyl)-2-oxoethyl)-l-azonia-bicyclo[2.2.2]octane bromide

(3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-l-( 2-(2-

C24

hydroxyphenyl)-2-oxoethyl)-l-azoniabicyclo[2.2.2]octane bromide

C26 (R)-quinuclidin-3-yl 2-(methoxycarbonylamino)-2-phenylacetate

(3R)-3-(2-(methoxycarbonylamino)-2-phenylacetoxy)-l -(2-0X0-2-

C27

phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide

C29 (R)-quinuclidin-3-yl 2-(benzyloxycarbonylamino)-2-phenylacetate

(continued) (3R)-3-(2-(benzyloxycarbonylamino)-2-phenylacetoxy)-l -(2-0X0-2-

C31

phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide

C32 (R)-quinuclidin-3-yl 2-phenyl-2-(vinyloxycarbonylamino)acetate

(3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2-

C33

(vinyloxycarbonylamino)acetoxy)-l-azoniabicyclo[2.2.2]octane bromide

C34 (R)-quinuclidin-3-yl 2-(ethoxycarbonylamino)-2-phenylacetate

(3R)-3-(2-(ethoxycarbonylamino)-2-phenylacetoxy)-l -(2-0X0-2-

C35

phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide

(R)-quinuclidin-3-yl 2-((2-methoxyethoxy)carbonylamino)-2-

C36

phenylacetate

(3R)-3-(2-((2-methoxyethoxy)carbonylamino)-2-phenylacetoxy)- l-(2-

C37

oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane bromide

C38 (R)-quinuclidin-3-yl 2-(cyclohexyloxycarbonylamino)-2-phenylacetate

(3R)-3-(2-(cyclohexyloxycarbonylamino)-2-phenylacetoxy)-l -(2-oxo-

C39

2-phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide

C41 (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-p-tolylacetate

(3R)-3-(2-(tert-butoxycarbonylamino)-2-p-tolylacetoxy)-l -(2-0X0-2-

C42

phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide

(R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(4-

C44

methoxyphenyl)acetate

(3R)-3-(2-(tert-butoxycarbonylamino)-2-(4-methoxyphenyl)acet oxy)-l-

C45

(2-oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane bromide

(R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(4-

C47

chlorophenyl)acetate

(3R)-3-(2-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)aceto xy)-l-

C48

(2-oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane bromide

(R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(4-

C50

fluorophenyl)acetate

(3R)-3-(2-(tert-butoxycarbonylamino)-2-(4-fluorophenyl)aceto xy)-l-(2-

C51

oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane bromide

(R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(3-

C53

fluorophenyl)acetate

(3R)-3-(2-(tert-butoxycarbonylamino)-2-(3-fluorophenyl)aceto xy)-l-(2-

C54

oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane bromide

(R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(2-

C56

fluorophenyl)acetate

(3R)-3-(2-(tert-butoxycarbonylamino)-2-(2-fluorophenyl)aceto xy)-l-(2-

C57

oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane bromide

(lR,3r,5S)-8-methyl-8-azabicyclo[3.2.1 ]octan-3-yl 2-(tert-

C58

butoxycarbonylamino)-2-phenylacetate

(lR,3r,5S)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)- 8-

C59

methyl-8-(2-oxo-2-phenylethyl)-8-azoniabicyclo[3.2.1]octane bromide

3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)-8,8-dime thyl-8-

C60

azoniabicyclo[3.2.1]octane iodide

(continued) (R)-l-methylpyrrolidin-3-yl 2-(tert-butoxycarbonylamino)-2-

C61

phenylacetate

(3R)-3 -(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)- 1 -methyl- 1 -(2-

C62

oxo-2-phenylethyl)pyrrolidinium bromide

(3R)-3 -(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)- 1 -methyl- 1 -(2-

C63

oxo-2-(thiophen-2-yl)ethyl)pyrrolidinium bromide

C64 (R)-quinuclidin-3-yl 2-phenyl-2-(phenylsulfonamido)acetate

(3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2-

C65

(phenylsulfonamido)acetoxy)-l-azoniabicyclo[2.2.2]octane chloride

(3R)-l-(2-oxo-2-(thiazol-2-yl)ethyl)-3-(2-phenyl-2-

C66 (phenylsulfonamido)acetoxy)-l-azoniabicyclo[2.2.2]octane 2,2,2- trifluoroacetate

(3R)-l-(2-oxo-2-(thiophen-2-yl)ethyl)-3-(2-phenyl-2-

C67 (phenylsulfonamido)acetoxy)-l-azoniabicyclo[2.2.2]octane 2,2,2- trifluoroacetate

(R)-quinuclidin-3-yl 2-(4-methoxyphenylsulfonamido)-2-phenylacetate

C68

hydrochloride

(3R)-3-(2-(4-methoxyphenylsulfonamido)-2-phenylacetoxy)-l-(2 -oxo-

C69

2-phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide

C70 (R)-quinuclidin-3-yl 2-(4-chlorophenylsulfonamido)-2-phenylacetate

(3R)-3-(2-(4-chlorophenylsulfonamido)-2-phenylacetoxy)- l -(2-0X0-2-

C71

phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide

(R)-quinuclidin-3-yl 2-(3,4-difluorophenylsulfonamido)-2-

C72

phenylacetate

(3R)-3-(2-(3,4-difluorophenylsulfonamido)-2-phenylacetoxy)- l-(2-oxo-

C73

2-phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(R)-quinuclidin-3-yl 2-(2,4-dimethylthiazole-5-sulfonamido)-2-

C74

phenylacetate

(3R)-3-(2-(2,4-dimethylthiazole-5-sulfonamido)-2-phenylaceto xy)-l-(2-

C75

oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

C76 (R)-quinuclidin-3-yl 2-(methylsulfonamido)-2-phenylacetate

(3R)-3-(2-(methylsulfonamido)-2-phenylacetoxy)-l -(2-0X0-2-

C77

phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide

(R)-quinuclidin-3-yl 2-phenyl-2-(2,2,2-

C78

trifluoroethylsulfonamido)acetate

(3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2-(2,2,2-

C79

trifluoroethyl sulfonamido)acetoxy)- 1 -azoni abi cy clo [2.2.2] octane bromi de

C80 (R)-quinuclidin-3-yl 2-phenyl-2-(phenylmethylsulfonamido)acetate

(3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2-

C81 (phenylmethylsulfonamido)acetoxy)- l-azoniabicyclo[2.2.2]octane

bromide

C83 (R)-quinuclidin-3-yl 2-(4-fluorophenyl)-2-(phenylsulfonamido)acetate

(3R)-3-(2-(4-fluorophenyl)-2-(phenylsulfonamido)acetoxy)- l -(2-0X0-2-

C84

phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(continued) C85 (R)-quinuclidin-3-yl 2-benzamido-2-phenylacetate

(3R)-3 -(2-benzamido-2-phenylacetoxy)- 1 -(2-oxo-2-phenylethyl)- 1 -

C86

azoniabicyclo[2.2.2]octane chloride

(3R)-3-(2-benzamido-2-phenylacetoxy)-l-(2-(4-fluorophenyl)-2 -

C87

oxoethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(3R)-3-(2-benzamido-2-phenylacetoxy)-l-(2-(4-hydroxyphenyl)- 2-

C88

oxoethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(3R)-3-(2-benzamido-2-phenylacetoxy)-l-(2-oxo-2-(thiazol-2-y l)ethyl)-

C89

l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(3R)-3-(2-benzamido-2-phenylacetoxy)-l-(2-oxo-2-(thiophen-3-

C90

yl)ethyl)- l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(3R)-3-(2-benzamido-2-phenylacetoxy)-l-(2-oxo-2-(thiophen-2-

C91

yl)ethyl)- l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

C92 (R)-quinuclidin-3-yl 2-acetamido-2-phenylacetate

(3R)-3 -(2-acetamido-2-phenylacetoxy)- 1 -(2-oxo-2-phenylethyl)- 1 -

C93

azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

C94 (R)-quinuclidin-3-yl 2-phenyl-2-pivalamidoacetate

(3R)- 1 -(2-oxo-2-phenylethyl)-3 -(2-phenyl-2-pivalamidoacetoxy)- 1 -

C95

azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(3R)-3-(2-(cyclopentanecarboxamido)-2-phenylacetoxy)-l -(2-0X0-2-

C96

phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2-(2-

C97 phenylacetamido)acetoxy)-l-azoniabicyclo[2.2.2]octane 2,2,2- trifluoroacetate

(3R)-3-(2-(3-ethoxy-3-oxopropanamido)-2-phenylacetoxy)-l -(2-0X0-2-

C98

phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(3R)-l-(2-oxo-2-phenylethyl)-3-(2-(2-phenoxyacetamido)-2-

C99

phenylacetoxy)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2-(thiophene-2-

CIOO carboxamido)acetoxy)-l-azoniabicyclo[2.2.2]octane 2,2,2- trifluoroacetate

(3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2-(thiazole-2-

ClOl carboxamido)acetoxy)-l-azoniabicyclo[2.2.2]octane 2,2,2- trifluoroacetate

(3R)-3-(2-(4-fluorobenzamido)-2-phenylacetoxy)-l -(2-0X0-2-

C102

phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(3R)-3-(2-(3-fluorobenzamido)-2-phenylacetoxy)-l -(2-0X0-2-

C103

phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(3R)-3-(2-(4-methylbenzamido)-2-phenylacetoxy)-l -(2-0X0-2-

C104

phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(3R)-3-(2-(4-methoxybenzamido)-2-phenylacetoxy)-l -(2-0X0-2-

C105

phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

C107 (R)-quinuclidin-3-yl 2-benzamido-2-(4-methoxyphenyl)acetate

(3R)-3-(2-benzamido-2-(4-methoxyphenyl)acetoxy)-l -(2-0X0-2-

C108

phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(continued) CllO (R)-quinuclidin-3-yl 2-benzamido-2-p-tolylacetate

(3R)-3 -(2-benzamido-2-p-tolylacetoxy)- 1 -(2-oxo-2-phenylethyl)- 1 - cm

azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

C113 (R)-quinuclidin-3-yl 2-benzamido-2-(4-chlorophenyl)acetate

(3R)-3-(2-benzamido-2-(4-chlorophenyl)acetoxy)-l -(2-0X0-2-

C114

phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(3R)-3-(2-(tert-butoxycarbonylamino)-2-(thiophen-2-yl)acetox y)-l-(2-

C116

oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane chloride

(3R)-3-(2-(tert-butoxycarbonylamino)-3-(lH-indol-3-yl)propan oyloxy)-

C118

l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide

((R)-3-((S)-3-(4-(benzyloxy)phenyl)-2-(tert-butoxycarbonylam ino)-

C120 propanoyloxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2] octane

bromide

(R)-3-((S)-2-(tert-butoxycarbonylamino)-3-(4-hydroxyphenyl)-

C122 propanoyloxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2] octane

bromide

(R)-3-((S)-2-(tert-butoxycarbonylamino)-3-phenylpropanoyloxy )-l -(2-oxo-

C124

2-phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide

(3R)-3-(2-(2-amino-2-phenylacetamido)-2-phenylacetoxy)-l -(2-0X0-2-

C127 phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate 2,2,2- trifluoroacetic acid

(3R)-3 -(2-benzamido-3 -methylbutanoyloxy)- 1 -(2-oxo-2-phenylethyl)- 1 -

C130

azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

(3R)-3-(2-benzamido-3 -methylbutanoyloxy)- 1 -(2-oxo-2-(thiophen-2-

C131

yl)ethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate

The compounds of formula (I) and (VI) may show at least one chiral center, when R ₃ is not H.

Further, depending on the meanings of Ri, R ₂ , R ₆ and R ₇ , it will be clear that additional asymmetric centers may be present in the compounds of formula (I) and (VI). Therefore, the invention also includes any of the optical stereoisomers, diastereoisomers and mixtures thereof, in any proportion.

In one of the preferred embodiment, the chiral center on rings (i), (iii), (iv), (v), (vii), (viii) and (ii), (vi) when m and n are different, shows a R configuration.

In the present invention, since the absolute configuration of the diastereomers is not always defined, they are indicated in the examples as diastereomer 1, 2 or mixtures of them.

The invention also provides pharmaceutical compositions of compounds of formula (I) or (VI) alone or in combination or in admixture with one or more pharmaceutically acceptable carriers and/or excipients.

The invention also provides pharmaceutical compositions suitable for administration by inhalation such as, for instance, inhalable powders, propellant-containing metering aerosols or propellant-free inhalable formulations.

The invention also provides compounds of formula (I) or (VI) for use as a medicament.

The invention also provides compounds of formula (I) or (VI) for use in the treatment of broncho-obstructive or inflammatory diseases, preferably asthma or chronic bronchitis or chronic obstructive pulmonary disease (COPD).

In a further aspect, the invention provides the use of the compounds of formula (I) or (VI) for the manufacture of a medicament for the prevention and/or treatment of broncho-obstructive or inflammatory diseases, preferably asthma or chronic bronchitis or chronic obstructive pulmonary disease (COPD).

The invention also provides a method for the prevention and/or treatment of broncho-obstructive or inflammatory diseases, preferably asthma or chronic bronchitis or chronic obstructive pulmonary disease (COPD), which comprises administering to a subject in need thereof a therapeutically effective amount of a compound of general formula (I) or (VI).

The invention also provides pharmaceutical compositions suitable for administration by inhalation, such as inhalable powders, propellant-containing metering aerosols or propellant-free inhalable formulations.

The invention also refers to a device which may be a single- or multi-dose dry powder inhaler, a metered dose inhaler and a soft mist nebulizer comprising the compounds of formula (I) or (VI). The invention also refers to a kit comprising the above pharmaceutical compositions in a suitable vial or container and a device which may be a single- or multi-dose dry powder inhaler, a metered dose inhaler and a soft mist nebulizer, adapted to hold the above vial or container.

The present invention is also directed to a process for the preparation of a compound of formula (I) or (VI) which comprises:

(a) the reaction of compounds of general formula (III)

with compounds of general formula (II)

to provide the corresponding compounds of general formula (IV)

(c) the coupling between compounds of general formula (IV) and (V)

(V)

HO.

R7

to obtain a compound of general formula (VI)

(d) the alkylation of compounds of general formula (VI) by alkylating agents of general formula (Y)

— (CH ₂ )p— P— (CH ₂ )q— W

(Y)

linked to a group A, which is a suitable leaving group, to obtain a compound of general formula (I), wherein p, P, q, W, Rl, R2, R3, R6 and R7 have the above reported meanings.

The present invention is also directed to a process for the preparation of a compound of formula (VI) which comprises:

(a) the treatment of the acid of formula (IV) with one or more equivalents of a condensing agent to obtain an activated intermediate;

(b) the reaction of the activated intermediate with alcohol of general formula (V).

The present invention is also directed to a process for the preparation of a compound of formula (VI) which comprises:

(a) the conversion of a compound of formula (IV) wherein K=OH into the corresponding acyl halide of formula (IV) wherein K=halide

(b) the reaction of the acyl halide of formula (IV) with a compound of formula (V).

The compounds of formula (I) or (VI) may be prepared according to known methods.

The starting materials for the preparation of the compounds of formula (I) or (VI), as well as any reactant of the process, are known or easily preparable according to known procedures.

The operative conditions that may be used in the process of the invention are described in more details below and are further reported in the following Scheme 1.

Coupling

Scheme 1

Procedure for the preparation of compounds of formula (VI) and (I) Compounds reported in the present invention can be most conveniently prepared starting from compounds of general formula (II), in which K may be either a hydroxyl group or a suitable protecting hydroxyl group (e.g. K= (C ₁ -C ₁₀ )alkoxy such as OMe). The compounds of general formula (II) can be reacted with compounds of general formula (III), in which z is a suitable leaving group such as an halide (i.e. chlorine, bromine, fluorine) or a oxygen substituted with another Rl-G group (e.g. when Rl is a tert-butyl group, G is a COO group and z is a -O-G-Rl group, compound (III) is di-tert-butyl dicarbonate or Boc anhydride; when Rl is methyl, G is CO and z is a -O-G-Rl group, compound (III) is acetic anhydride; when Rl is CF ₃ , G is SO ₂ and z is a -O-G-Rl group, compound (III) is trifluoromethanesulphonic anhydride). This reaction can be effected according to standard procedures reported in the literature. In a typical procedure, compounds of formula (III) are added to a solution of compounds of formula (II) in a suitable solvent (e.g. dichloromethane, ethyl acetate, tetrahydrofuran and water) to provide the corresponding compounds of general formula (IV). The reaction is conveniently promoted by a base such as triethylamine, pyridine, 4-dimethylaminopyridine and sodium hydroxide. This reaction is usually performed in a temperature range from 0°C to 130°C over a period of 30 minutes up to 74 hours. The reaction may be carried out under conventional heating (using an oil bath) or microwave heating. The reaction may be carried out in an open vessel or in a sealed tube.

Reagents of general formula (III) are commercially available or may be prepared according to standard procedures reported in literature. When z in compounds (III) is a hydroxyl group (z=OH), it may be either converted into a suitable leaving group (such as a halide or a oxygen substituted with another Rl-G group to form an anhydride) or condensed to compounds of formula (II) under standard amidation and peptide coupling conditions.

The coupling between compounds of general formula (IV) and (V) may be conducted in several ways (a survey of the suitable reactions is given by Carey, F.A. and Sundeberg, R.J. Advanced Organic Chemistry, Third Edition (1990), Plenum Press, New York and London, pg 145).

In particular, in case K is a protecting hydroxyl group, the protecting group has to be removed before the coupling with (V). For instance, if K=OMe, hydrolysis of ester moiety may be performed treating the compound (IV) wherein K=OMe with a suitable aqueous base selected from sodium, lithium and potassium hydroxide in the suitable solvents (e.g. tetrahydrofuran, dioxane etc). The reaction proceeds at room temperature (RT), over a period of 1 hour up to 36 hours.

Alternative one - In a typical procedure, compounds (VI) may be prepared by condensation between compounds of formula (V) and (IV) wherein K=OH, under standard amidation and peptide coupling conditions. For instance, treatment of compound (IV) with one or more equivalents of a commercially available condensing agent such as a carbodiimide (e.g. Ν,Ν'-Dicyclohexylcarbodiimide (DCC) and the like) for example in the presence of N-hydroxybenzotriazole (HOBt) followed by reaction of the activated intermediate with alcohol (V), results in the formation of compounds of formula (VI). An organic base such as triethylamine or 4-dimethylaminopyridine may be also present in the reaction mixture. The activated intermediate may be either isolated, or pre-formed or generated in situ. Suitable solvents for the coupling include, but are not limited to, halocarbon solvents (e.g. dichloromethane), tetrahydrofuran, dioxane and acetonitrile. The reaction proceeds at temperature range from 0°C up to 170°C, for a time in the range of about 1 hour up to 72 hours. The reaction may be carried out under conventional heating (using an oil bath) or under microwave irradiation. The reaction may be conducted either in an open vessel or in a sealed tube.

Alternative two - In some embodiments of the present invention, a compound of formula (IV) wherein K=OH, is first converted into the corresponding acyl halide (IV) wherein K=halide. This activation may be effected according to one of the standard procedures reported in the literature. For instance, treatment of acid (IV) wherein K=OH, with one or more equivalents of oxalyl chloride in the presence of a catalytic amount of dimethylformamide (DMF) in a halocarbon solvent, such as dichloromethane, at temperature ranging form 0°C to 35°C, affords the required acyl chloride (IV) wherein K=C1.

Alcohol (V) is then reacted with the acyl halide (IV), using known methods. The reaction may be promoted by a base such as triethylamine, pyridine and 4-dimethylaminopyridine, in a suitable solvent (e.g. dichloromethane). This reaction is performed in a temperature range from 0°C to 130°C over a period of 1 hour up to 74 hours. The reaction may be conducted under conventional heating (using an oil bath) or under microwave heating. The reaction may be carried out in an open vessel or in a sealed tube.

Alternative three - Alternatively, acylation of alcohol (V) to give compounds of general formula (VI) may be accomplished using procedures which convert in situ the acid (IV) wherein K=OH, into the corresponding acyl halides. For example, alcohols (V) are reacted with acids (IV) wherein K=OH, in presence of triphenylphosphine and a halocarbon solvent such as carbon tetrachloride or dichloromethane, at about RT, in a maximum period of time of 16 hours (Lee, J.B. J.Am.Chem.Soc, 1966, 88, 3440).

Alternative four - In another process for the preparation of the compounds of the present invention, acid (IV) wherein K=OH, may be activated with other commercially available activating agents such as bromotripyrrolidinophosphonium hexafluorophosphate (PyBrOP) or carbonylimidazole, in the suitable solvent (e.g. dichloromethane, tetrahydrofuran and DMF), at about RT. Subsequent reaction of the activated intermediate with alcohol (V) provides the desired compound of formula (VI). The reaction may also require the use of an organic base such as diisopropylethylamine and the like and usually proceeds at about RT.

Alternative five -In another process for the preparation of the compounds of the present invention, compounds (VI) can be efficiently prepared by the condensation between acids (IV) wherein K=OH and alcohol

(V) under typical Mitsunobu conditions (Kumara Swamy, K.C., Chem. Rev. 2009, 109, 2551-2651). For example, acids (IV) and alcohol (V) are reacted in presence of a phosphine (e.g. triphenylphosphine) and an azadicarboxylate ester (e.g. diethyl azodicarboxylate or diisopropyl azodicarboxylate) in an aprotic solvent such as tetrahydrofuran. The reaction typically proceeds at temperature range from 0°C up to 100°C, for a time in the range of about 30 minutes up to 72 hours.

In some embodiments of the present invention, group Rl-G- in compounds of general formula (VI) can be most conveniently used as a protecting group and can be removed to obtain compounds of general formula (VIII). These protecting groups are selected, manipulated and removed according to standard methods of organic synthesis (Green T.W. and Wuts P.G.M. (1991) Protecting Groups in Organic Synthesis, John Wiley et Sons). For instance, if Rl in compounds (VI) is tert-butyl and G is O(CO), the tert- butyloxycarbonyl (boc) protecting group can be cleaved by treating compounds of general formula (VI) with a protic acid such as hydrochloric acid, trifluoroacetic acid and the like. Suitable solvents for deprotection include, but are not limited to, 1,4-dioxane, tetrahydrofuran, dichloromethane and methanol. Water may be also present in the reaction mixture. The reaction proceeds at temperature range from 0°C up to 80°C, for a time in the range of few minutes up to 72 hours.

Compounds of general formula (VIII) may be then coupled to compounds of general formula (III) to yield compounds (VI), applying known procedures. For instance, the conditions used to perform the coupling may be selected among those described to produce the coupling between compound (II) and (III) in Scheme 1.

Compound of general formula (VI) can be obtained either as single diastereoisomer or as a mixture of diastereoisomers. For instance, in the case R7 is a group of formula (V), the alcohol features either a R or a S configuration. If the R-enantiomer is used, the compound of formula (VI) can be obtained in S-R configuration, in R-R configuration or as a mixture of diastereoisomers (R-R and S-R configuration).

The mixture of diastereoisomers may be converted to compounds of formula (I) of Scheme 1 or resolved to give the two single diastereoisomers, which in turn may be converted to compounds of formula (I) of Scheme 1. This separation can be accomplished using known procedures. These procedures include, but are not limited to, chromatography purification, preparative HPLC purification and crystallization. For example, the two diastereoisomers can be separated by flash chromatography on silica gel eluting with suitable solvents or mixture of solvents such as DCM and methanol and the like. In another process of the present invention separation of diastereoisomers may be obtained using a column filled with a chiral stationary phase, for example Chiralpack® AY or Chiralcel OD® or Chiralcel OZ®, and eluting, for example, with acetonitrile and/or with mixtures of acetonitrile and an alcohol. Alternatively, the separation of diastereoisomers may be most conveniently achieved by crystallization from a suitable solvent (e.g. ethyl ether and acetone), as a free base or after the formation of a suitable salt (e.g. D-tartaric acid)).

The alkylation of compounds of general formula (VI) by alkylating agents of general formula (Y), linked to a suitable leaving group A selected from the group consisting of halide (i.e. bromine, iodine, chlorine) and sulfonate ester (i.e. tosylate, triflate, mesylate), gives the compounds of general formula (I). This kind of reaction is widely described in literature under several different conditions, for instance, the reaction may be performed neat or in a suitable solvent selected from acetonitrile, ethyl acetate, DMF, DMSO and tetrahydrofuran. The reaction typically proceeds at temperatures from 0°C up to 170°C, for a time in the range of few minutes up to 72 hours. The reaction may be carried out under conventional heating (using an oil bath) or under microwave irradiation. The reaction may be conducted either in an open vessel or in a sealed tube.

Compounds of general formula (I) in Scheme 1 can be either considered as final products or can be further reacted to prepare other compounds of general formula (I). Thus, a moiety of Rl, R2, R3 or R6 group in general formula (I) can undergo oxidation, reduction or cleavage reactions (e.g. to remove a needed protecting group) to afford other final compounds of general formula (I).

The present invention also provides pharmaceutical compositions of compounds of formula (I) or (VI) in admixture with one or more pharmaceutically acceptable carriers, for example those described in Remington's Pharmaceutical Sciences Handbook, XVII Ed., Mack Pub., N.Y., U.S.A.

Administration of the compounds of the invention may be accomplished according to patient needs, for example, orally, nasally, parenterally (subcutaneously, intravenously, intramuscularly, intrasternally and by infusion), by inhalation, rectally, vaginally, topically, locally, transdermally, and by ocular administration.

Various solid oral dosage forms can be used for administering compounds of the invention including such solid forms as tablets, gelcaps, capsules, caplets, granules, lozenges and bulk powders. The compounds of the present invention can be administered alone or combined with various known pharmaceutically acceptable carriers, diluents (such as sucrose, mannitol, lactose, starches) and excipients, including but not limited to suspending agents, solubilizers, buffering agents, binders, disintegrants, preservatives, colorants, flavors, lubricants and the like. Time release capsules, tablets and gels are also advantageous. Various liquid oral dosage forms can also be used for administering the compounds of the invention, including aqueous and non-aqueous solutions, emulsions, suspensions, syrups, and elixirs. Such dosage forms can also contain suitable known inert diluents such as water, excipients such as preservatives, wetting agents, sweeteners, flavors, as well as agents for emulsifying and/or suspending the compounds of the invention. The compounds of the invention may be injected, for example, intravenously, in the form of an isotonic sterile solution. Other preparations are also possible.

Suppositories for rectal administration of the compounds of the present invention can be prepared by mixing the compound with a suitable excipient such as cocoa butter, salicylates and polyethylene glycols.

Formulations for vaginal administration can be in the form of cream, gel, paste, foam, or spray formula containing, in addition to the active ingredient, such suitable carriers as are known in the art.

For topical administration the pharmaceutical composition can be in the form of creams, ointments, liniments, lotions, emulsions, suspensions, gels, solutions, pastes, powders, sprays, and drops suitable for administration to the skin, eye, ear or nose. Topical administration may also involve transdermal administration via means such as transdermal patches.

For the treatment of the diseases of the respiratory tract, the compounds according to the invention are preferably administered by inhalation.

Inhalable compositions include inhalable powders, propellant-containing metering aerosols or propellant-free inhalable formulations.

For administration as a dry powder, known single- or multi-dose inhalers may be utilized. In that case the powder may be filled in gelatine, plastic or other capsules, cartridges or blister packs or in a reservoir.

A diluent or carrier, generally non-toxic and chemically inert to the compounds of the invention, e.g. lactose or any other additive suitable for improving the respirable fraction, may be added to the powdered compounds of the invention.

Inhalation aerosols containing propellant gas such as hydrofluoroalkanes may contain the compounds of the invention either in solution or in dispersed form. The propellant-driven formulations may also contain other ingredients such as co-solvents, stabilizers and optionally other excipients.

The propellant-free inhalable formulations comprising the compounds of the invention may be in form of solutions or suspensions in an aqueous, alcoholic or hydroalcoholic medium and they may be delivered by jet or ultrasonic nebulizers or by soft-mist nebulizers.

The compounds of the invention may be administered as the sole active agent or in combination with other pharmaceutical active ingredients including those currently used in the treatment of respiratory disorders, e.g. beta2-agonists, corticosteroids, P38 MAP kinase inhibitors, IKK2 inhibitors,

HNE inhibitors, PDE4 inhibitor, leukotriene modulators, NSAIDs and mucus regulators.

The invention also provides combinations of a compound of formula (I) or (VI) with a 2-agonist selected from the group consisting of GSK-642444, indacaterol, milveterol, arformoterol, salbutamol, levalbuterol, terbutaline, AZD-3199, BI-1744-CL, LAS- 100977, bambuterol, isoproterenol, procaterol, clenbuterol, reproterol, fenoterol and ASF- 1020.

The invention also provides combinations of a compound of formula (I) or (VI) with a corticosteroid selected from the group consisting of propionate, ciclesonide, mometasone furoate and budesonide.

The invention also provides combinations of a compound of formula (I) or (VI) with a P38 inhibitor selected from the group consisting of semapimod, talmapimod, pirfenidone, PH-797804, GSK-725, minokine and losmapimod.

The invention also provides combinations of a compound of formula (I) or (VI) with an IKK2 inhibitor.

The invention also provides combinations of a compound of formula (I) or (VI) with a HNE inhibitor selected from the group consisting of AAT, ADC-7828, Aeriva, TAPI, AE-3763, KRP- 109, AX-9657, POL-6014, AEPv-002, AGTC-0106, respriva, AZD-9668, zemaira, AAT IV, PGX-100, elafin, SPHD-400, prolastin C and prolastin inhaled.

The invention also provides combinations of a compound of formula (I) or (VI) with a PDE4 inhibitor selected from the group consisting of AN-2728, AN-2898, CBS-3595, apremilast, ELB-353, KF-66490, K-34, LAS-37779, IBFB-21 1913, AWD- 12-281, cipamfylline, cilomilast, roflumilast, BAY19-8004 and SCH-351591, AN-6415, indus-82010, TPI-PD3, ELB-353, CC-11050, GSK-256066, oglemilast, OX-914, tetomilast, MEM- 1414 and

The invention also provides combinations of a compound of consisting of montelukast, zafirlukast and pranlukast.

The invention also provides combinations of a compound of formula (I) or (VI) with a NSAID selected from the group consisting of ibuprofen and ketoprofen.

The invention also provides combinations of a compound of formula (I) (VI) with a mucus regulator selected from the group consisting of INS-37217, diquafosol, sibenadet, CS-003, talnetant, DNK-333, MSI-1956 and gefitinib.

The dosages of the compounds of the present invention depend upon a variety of factors including the particular disease to be treated, the severity of the symptoms, the route of administration, the frequency of the dosage interval, the particular compound utilized, the efficacy, toxicology profile, and pharmacokinetic profile of the compound. Advantageously, the compounds of formula (I) or (VI) can be administered for example, at a dosage comprised between 0.001 and 1000 mg/day, preferably between 0.1 and 500 mg/day.

When the compounds of formula (I) or (VI) are administered by inhalation route, they are preferably given at a dosage comprised between 0.001 and 500 mg/day, preferably between 0.1 and 200 mg/day.

The compounds of formula (I) or (VI) may be administered for the prevention and/or treatment of any disease wherein M3 antagonists are active. Said disease include: diseases involving inflammation such as asthma and COPD, acute rhinitis; diseases of the gastrointestinal tract such as peptic ulcer; diseases of the cardiovascular system such as acute myocardial infarction; diseases of the genitourinary tract such as renal colic; anticholinesterase and mushroom poisoning; uses in anesthesia; uses in ophthalmology.

They also include neurological and psychiatric disorders such as Parkinsonism and motion sickness.

Preferably, the compounds of formula (I) or (VI) may be administered for the prevention and/or treatment of respiratory diseases such as from mild to acute severe conditions of asthma and COPD.

Other respiratory diseases include bronchitis, bronchiolitis, bronchiectasis, acute nasopharyngitis, acute and chronic sinusitis, maxillary sinusitis, pharyngitis, tonsillitis, laryngitis, tracheitis, epiglottitis, croup, chronic disease of tonsils and adenoids, hypertrophy of tonsils and adenoids, peritonsillar abscess, rhinitis, abscess or ulcer and nose, pneumonia, viral and bacterial pneumonia, bronchopneumonia, influenza, extrinsic allergic alveolitis, coal workers' pneumoconiosis, asbestosis, pneumoconiosis, pneumonopathy, respiratory conditions due to chemical fumes, vapors and other external agents, emphysema, pleurisy, pneumothorax, abscess of lung and mediastinum, pulmonary congestion and hypostasis, postinflammatory pulmonary fibrosis, other alveolar and parietoalveolar pneumonopathy, idiopathic fibrosing alveolitis, Hamman-Rich syndrome, atelectasis, ARDS, acute respiratory failure, mediastinitis.

The invention will be further described by the following examples.

I = intermediates

C = compounds.

EXAMPLE 1

Preparation of (R)-3-((R)-2-(tert-butoxycarbonylamino)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane bromide (Diastereoisomers 1 of C2)

diasteroisomer 1 of C1 diasteroisomer 1 of C2

Scheme 2

Preparation of (R)-((R)-quinuclidin-3-yl) 2-(tert- butoxycarbonylamino)-2-phenylacetate (Diastereoisomer 1 of CI)

A mixture of (R)-2-(tert-butoxycarbonylamino)-2-phenylacetic acid (1.00 g, 3.98 mmol), (R)-quinuclidin-3-ol (0.51 g, 3.98 mmol), lH-benzo[d][ l,2,3]triazol-l-ol (0.64 g, 4.78 mmol) and DCC (0.98 g, 4.78 mmol) was stirred at RT overnight. Then THF was removed in vacuum and the residue was partitioned between EtOAc and 2M K ₂ CO ₃ . The organic phase was dried over Na ₂ SO ₄ , filtered and evaporated to dryness. The crude was purified by flash chromatography (DCM/MeOH= 98/2 to 95/5) to obtain (R)-((R)-quinuclidin-3-yl) 2-(tert-butoxycarbonylamino)-2-phenylacetate (612 mg; 43% yield). 1H NMR (300 MHz, DMSO-<¾) δ ppm 7.74 (d, 1 H), 7.17 - 7.52 (m, 5 H), 5.19 (d, 1 H), 4.58 - 4.82 (m, 1 H), 2.99 (ddd, 1 H), 2.55 - 2.69 (m, 3 H), 2.32 - 2.46 (m, 1 H), 2.18 (d, 1 H), 1.79 - 1.98 (m, 1 H), 1.44 - 1.71 (m, 3 H), 1.40 (s, 9 H), 1.11 - 1.33 (m, 1 H);

LC-MS (ESI POS): 361.4 (MH+).

Preparation of (R)-3-((R)-2-(tert-butoxycarbonylamino)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane bromide (Diastereoisomers 1 of C2)

2-Bromo-l-phenylethanone (27.6 mg, 0.14 mmol) was added to a solution of (R)-((R)-quinuclidin-3-yl) 2-(tert-butoxycarbonylamino)-2- phenylacetate (Diastereoisomer 1 of CI) (50 mg, 0.14 mmol) in EtOAc (3 ml). The reaction was stirred at RT overnight. The precipitate was collected by suction filtration and washed with Et ₂ O to obtain (R)-3-((R)-2-(tert- butoxycarbonylamino)-2-phenylacetoxy)-l-(2-oxo-2-phenylethyl )-l- azoniabicyclo[2.2.2]octane bromide (65 mg; 84% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 7.92 - 8.03 (m, 2 H), 7.89 (d, 1 H), 7.70 - 7.80 (m, 1 H), 7.56 - 7.68 (m, 2 H), 7.30 - 7.52 (m, 5 H), 5.30 (d, 1 H), 5.17 - 5.24 (m, 1 H), 5.15 (s, 2 H), 4.02 - 4.19 (m, 1 H), 3.42 - 3.83 (m, 5 H), 2.30 - 2.42 (m, 1 H), 1.75 - 2.12 (m, 4 H), 1.42 (s, 9 H);

LC-MS (ESI POS): 479.09 (M+).

EXAMPLE 2

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane chloride (C2)

Scheme 3

Preparation of 2-(tert-butoxycarbonylamino)-2-phenylacetic acid (13):

A mixture of 2-amino-2-phenylacetic acid (2.00 g, 13.2 mmol) and di-tert-butyl dicarbonate (3.47 g, 15.9 mmol) in sodium hydroxide (50 ml, 100 mmol) and acetone (50 ml) was stirred at RT for 1 hr. Acetone was removed under reduced pressure, the aqueous phase was acidified to pH about 5 with HCl and extracted twice with EtOAc. The organic phase was dried over Na ₂ SO ₄ , filtered and evaporated to dryness to provide 2-(tert-butoxycarbonylamino)-2-phenylacetic acid (1.42 g; 43% yield). The compound was used in the next step without any further purification.

Preparation of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)- 2-phenylacetate (CI)

A mixture of 2-(tert-butoxycarbonylamino)-2-phenylacetic acid (13) (2.00 g, 7.96 mmol), (R)-quinuclidin-3-ol (1.21 g, 9.55 mmol), HOBt (1.46 g, 9.55 mmol) and DCC (1.97 g, 9.55 mmol) in dry THF (70 ml) was stirred at RT overnight. Then THF was evaporated and the crude was taken up with DCM and washed twice with 2M K ₂ CO ₃ and brine. The organic phase was dried over Na ₂ SO ₄ , filtered and evaporated to dryness. The crude was purified by flash chromatography (DCM/MeOH=95/5) to provide (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-phenylacetate (1.68 g; 58.5% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm

Diastereoisomer 1 : 7.73 (d, 1 H), 7.12 - 7.54 (m, 5 H), 5.19 (d, 1 H), 4.52 - 4.84 (m, 1 H), 2.99 (ddd, 1 H), 2.54 - 2.70 (m, 3 H), 2.31 - 2.47 (m, 1 H), 2.12 - 2.24 (m, 1 H), 1.84 - 1.92 (m, 1 H), 1.47 - 1.71 (m, 2 H), 1.40 (s, 9 H), 1.06 - 1.36 (m, 2 H);

Diastereoisomer 2: 7.73 (d, 1 H), 7.12 - 7.54 (m, 5 H), 5.19 (d, 1 H), 4.52 - 4.84 (m, 1 H), 2.99 (ddd, 1 H), 2.54 - 2.70 (m, 5 H), 1.69 - 1.79 (m, 1 H), 1.47 - 1.71 (m, 2 H), 1.40 (s, 9 H), 1.06 - 1.36 (m, 2 H);

LC-MS (ESI POS): 361.16 (MH+).

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane chloride (C2):

2-Chloro-l-phenylethanone (30.0 mg, 0.19 mmol) was added to a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-phenylacetate (CI) (70.0 mg, 0.19 mmol) in EtOAc (2 ml). The reaction was stirred at RT for 24h then Et ₂ O (1 ml) was added and the reaction was sonicated. The solid was collected by suction filtration to obtain (3R)-3-(2-(tert- butoxycarbonylamino)-2-phenylacetoxy)-l-(2-oxo-2-phenylethyl )-l- azoniabicyclo[2.2.2]octane chloride (54.6 mg; 55% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 7.94 - 8.05 (m, 2 H), 7.89 (d, 1 H), 7.70 - 7.82 (m, 1 H), 7.54 - 7.70 (m, 2 H), 7.28 - 7.53 (m, 5 H), 5.27 - 5.37 (m, 1 H), 5.08 - 5.27 (m, 2 H), 3.95 - 4.28 (m, 1 H), 3.44 - 3.88 (m, 5 H), 2.19 and 2.37 (br. s., 1 H), 1.52 - 2.12 (m, 4 H), 1.41 (s, 9 H);

LC-MS (ESI POS): 479.21 (M+).

The compounds listed in Table 1 were obtained as previously described for C2, starting from compound CI and the suitable commercially available alkylating agents.

Table 1

(continued)

(continued)

(continued)

EXAMPLE 3

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2- phenylacetoxy)-l-(2-oxo-2-(thiophen-3-yl)ethyl)-l-azoniabicy clo [2.2.2]- octane bromide (C17)

Scheme 4

2-Bromo- l-(thiophen-3-yl)ethanone (39.8 mg, 0. 19 mmol) was added to a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2- phenylacetate (C I) (70.0 mg, 0. 19 mmol) in ethyl acetate (2 ml). The reaction was stirred at RT overnight. Et ₂ O ( 1 ml) was added and the solid was collected by suction filtration. The compound was further purified by flash chromatography (DCM/MeOH=95/5) to obtain (3R)-3-(2-(tert- butoxycarbonylamino)-2-phenylacetoxy)- l-(2-oxo-2-(thiophen-3-yl)ethyl)- l- azoniabicyclo[2.2.2]octane bromide (55.6 mg; 51% yield).

1H NMR (300 MHz, DMSO-< ) δ ppm 8.60 - 8.65 (m, 1 H) 7.88 (d, 1 H) 7.75 and 7.73 (dd, 1 H) 7.5 1 - 7.61 (m, 1 H) 7.27 - 7.50 (m, 5 H) 5.10 - 5.39 (m, 2 H) 4.93 - 5.08 (m, 1 H) 4.02 - 4.22 (m, 1 H) 3.43 - 3.79 (m, 5 H) 2.30 - 2.41 (m, 1 H) 1.85 - 2. 1 1 (m, 5 H) 1.41 (s, 9 H);

LC-MS (ESI POS): 485.06 (M+). EXAMPLE 4

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2- ylacetoxy)-l-methyl-l-azoniabicyclo[2.2.2]octane iodide (C18)

Scheme 5

Methyl iodide (8.6 μΐ, 0.14 mmol) was added to a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-phenylacetate (CI) (50 mg, 0.14 mmol) in ethyl acetate (2 ml) and the reaction was stirred at RT overnight. Et ₂ O (1 ml) was added and the precipitate was collected by suction filtration and dried under vacuum at 40°C. The product was further purified by preparative HPLC to obtain (3R)-3-(2-(tert-butoxycarbonylamino)-2- phenylacetoxy)-l-methyl- l-azoniabicyclo[2.2.2]octane iodide (24.2 mg; 35% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 7.85 (d, 1 H) 7.20 - 7.55 (m, 5 H) 5.26 and 5.31 (d, 1 H) 4.93 - 5.18 (m, 1 H) 3.83 (ddd, 1 H) 3.30 - 3.53 (m, 3 H) 3.09 - 3.27 (m, 1 H) 3.16 (dt, 1 H) 2.93 and 2.95 (s, 3 H) 2.04 - 2.14 and 2.25 - 2.33 (m, 1 H) 1.52 - 2.03 (m, 4 H) 1.40 (s, 9 H);

LC-MS (ESI POS): 375.20 (M+).

The compound listed in Table 2 was obtained as previously described for C18, starting from intermediate CI and (3-bromopropoxy)benzene. Table 2

EXAMPLE 5

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2- phenylacetoxy)-l-(2-(2-methoxyphenyl)-2-oxoethyl)-l- azoniabicyclo[2.2.2]octane bromide (C20)

Scheme 6

2-Bromo-l-(2-methoxyphenyl)ethanone (45.8 mg, 0.20 mmol) was added to a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2- phenylacetate (CI) (72.0 mg, 0.20 mmol) in acetonitrile (3 ml). The reaction was stirred at RT overnight. The product was collected by suction filtration to provide (3R)-3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)- l-(2-(2- methoxyphenyl)-2-oxoethyl)- l-azoniabicyclo[2.2.2]octane bromide (1 12 mg; 95% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 7.88 (d, 1 H), 7.76 - 7.84 (m, 1 H), 7.64 - 7.75 (m, 1 H), 7.30 - 7.54 (m, 5 H), 7.23 - 7.30 (m, 1 H), 7.03 - 7. 19 (m, 1 H), 5.24 - 5.36 (m, 1 H), 5. 1 1 - 5.24 (m, 1 H), 4.89 (s, 2 H), 4.03 - 4.23 (m, 1 H), 3.95 and 3.96 (s, 3 H), 3.45 - 3.84 (m, 5 H), 2.13 - 2.23 and 2.3 1 - 2.42 (m, 1 H), 1.51 - 2. 12 (m, 4 H), 1.41 (s, 9 H);

LC-MS (ESI POS): 509.33 (M+).

The compound listed in Table 3 was obtained as previously described for C20, starting from intermediate C I and 3-(chloromethyl)-5-phenyl- 1,2,4- oxadiazole.

Table 3

EXAMPLE 6

Preparation of (3R)-l-(2-tert-butoxy-2-oxoethyl)-3-(2-(tert- butoxycarbonylamino)-2-phenylacetoxy)-l-azoniabicyclo [2.2.2]octane bromide(C22)

Scheme 7

Tert-butyl 2-bromoacetate (29.5 μΐ, 0.20 mmol) was added to a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-phenylacetate (C I) (72 mg, 0.20 mmol) in acetonitrile (3 ml). The reaction was stirred at RT overnight. The product was collected by suction filtration and then purified by flash chromatography (DCM/MeOH=98/2 to 95/5) to provide (3R)- l-(2-tert- butoxy-2-oxoethyl)-3-(2-(tert-butoxycarbonylamino)-2-phenyla cetoxy)- l- azoniabicyclo[2.2.2]octane bromide (97 mg; 87% yield).

1H NMR (300 MHz, DMSO-< ) δ ppm 7.87 (d, 1 H), 7.20 - 7.54 (m, 5 H), 5.23 - 5.44 (m, 1 H), 5.08 - 5.22 (m, 1 H), 4.21 and 4.23 (s, 2 H), 3.88 - 4.08 (m, 1 H), 3.37 - 3.72 (m, 5 H), 2.09 - 2.22 and 2.30 - 2.40 (m, 1 H), 1.76 - 2.08 (m, 4 H), 1.46 and 1.48 (s, 9 H), 1.41 (s, 9 H);

LC-MS (ESI POS): 475.33 (M+).

The compounds listed in Table 4 were obtained as previously described for C22, starting from intermediate C I and 2-bromo- l-(2- nitrophenyl)ethanone or 2-bromo- l-(2-hydroxyphenyl)ethanone. Table 4

EXAMPLE 7

Preparation of (3R)-3-(2-(methoxycarbonylamino)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane

Scheme 8

Preparation of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125)

To a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2- phenylacetate (CI) (1.25 g, 3.47 mmol) in THF (20 ml), 37% hydrogen chloride (2.00 ml, 24.3 mmol) was added dropwise. The reaction was stirred at RT for 15h. The solvent was evaporated to obtain (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (1.16 g; quantitative yield) as a solid. The compound was used in the next step without any further purification. Preparation of (R)-quinuclidin-3-yl 2-(methoxycarbonylamino)-2- phenylacetate (C26)

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (150 mg, 0.45 mmol) in DCM (5 ml), triethylamine (188 μΐ, 1.35 mmol) and methyl carbonochloridate (41.7 μΐ, 0.54 mmol) were added. The reaction was stirred at RT for 4h and then the solvent was evaporated. The residue was taken up with EtOAc and washed with water and brine. The organic phase was dried over Na ₂ SO ₄ and evaporated to obtain (R)-quinuclidin-3-yl 2-(methoxycarbonylamino)-2-phenylacetate (58.0 mg; 40% yield). The compound was used in the next step without any further purification.

Preparation of (3R)-3-(2-(methoxycarbonylamino)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane bromide (C27)

To a solution of (R)-quinuclidin-3-yl 2-(methoxycarbonylamino)-2- phenylacetate (C26) (58.0 mg, 0.18 mmol) in EtOAc (3 ml) and CH ₃ CN (1 ml), 2-bromo-l-phenylethanone (39.9 mg, 0.20 mmol) was added. The reaction was stirred at RT for 15h and then the solvent was evaporated. The crude was purified by flash chromatography (DCM/MeOH= 95/5 to 92/8) and the resulting product was triturated with z-Pr ₂ O to obtain (3R)-3-(2- (methoxycarbonylamino)-2-phenylacetoxy)- 1 -(2-oxo-2-phenylethyl)- 1 - azoniabicyclo[2.2.2]octane bromide (55.2 mg; 58.6% yield).

1H NMR (300 MHz, DMSO-d6) δ ppm 8.17 - 8.30 (m, 1 H) 7.98 (dd, 2 H) 7.71 - 7.84 (m, 1 H) 7.61 (td, 2 H) 7.30 - 7.52 (m, 5 H) 5.36 (dd, 1 H) 5.20 - 5.30 (m, 1 H) 5.16 (d, 2 H) 4.01 - 4.23 (m, 1 H) 3.61 (s, 3 H) 3.45 - 3.81 (m, 5 H) 2.39 (s, 1 H) 1.90 - 2.11 (m, 3 H) 1.54 - 1.71 (m, 1 H);

LC-MS (ESI POS): 437.12 (M+). EXAMPLE 8

Preparation of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate (130)

Scheme 9

Preparation of 2-(benzyloxycarbonylamino)-2-phenylacetic acid

(128)

To a solution of 2-amino-2-phenylacetic acid (500 mg, 3.31 mmol) in

2N sodium hydroxide (1.65 ml, 3.31 mmol) stirred at 0°C, benzyl carbonochloridate (512 μΐ, 3.64 mmol) and 2N sodium hydroxide (1.82 ml,

3.64 mmol) were simultaneously added dropwise from two different syringes.

The reaction was stirred at RT for 45 minutes and a precipitate appeared.

Water was added and the solution was extracted with Et ₂ O. The aqueous phase was acidified with IN HCl and the desired product was extracted again with Et ₂ O. The combined organic phases were dried over Na ₂ SO ₄ , filtered and evaporated to obtain 2-(benzyloxycarbonylamino)-2-phenylacetic acid

(855 mg; 91% yield).

Preparation of (R)-quinuclidin-3-yl 2-(benzyloxycarbonylamino)-2- phenylacetate (C29)

To a solution of 2-(benzyloxycarbonylamino)-2-phenylacetic acid (128) (855 mg, 3.00 mmol) in THF (20 ml), (R)-quinuclidin-3-ol (457 mg, 3.60 mmol), N,N'-methanediylidenedicyclohexanamine (742 mg, 3.60 mmol) and lH-benzo[d][l,2,3]triazol-l-ol (486 mg, 3.60 mmol) were added. The reaction was stirred at RT for 15h then the solvent was evaporated. The residue was taken up with DCM, the insoluble solid was filtered off and the clear solution was washed twice with Na ₂ CO ₃ and brine, dried over Na ₂ SO ₄ and evaporated. The resulting crude is purified by silica gel flash chromatography (DCM/MeOH=9/l) to obtain (R)-quinuclidin-3-yl 2-(benzyloxy-carbonylamino)-2-phenylacetate (925 mg; 78% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 8.26 (d, 1 H), 7.19 - 7.54 (m, 10 H), 5.28 and 5.29 (d, 1 H), 5.10 (d, 1 H), 5.05 (d, 1 H), 4.59 - 4.80 (m, 1 H), 2.95 - 3.07 (m, 1 H), 2.09 - 2.71 (m, 5 H), 1.70 - 1.80 and 1.81 - 1.99 (m, 1 H), 1.01 - 1.70 (m, 4 H).

Preparation of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (130)

A solution of (R)-quinuclidin-3-yl 2-(benzyloxycarbonylamino)-2- phenylacetate (C29) (100 mg, 0.25 mmol) in MeOH (7 ml) and 37% hydrogen chloride (20.8 μΐ, 0.25 mmol) was stirred at RT under hydrogen atmosphere (25 psi) for 3h in a Parr apparatus, in presence of palladium on activate carbon (10 mg, 9.40 μιηοΐ). The catalyst was filtered off and the solvent was evaporated. The resulting oil was purified by SCX cartridge eluting with MeOH and then MeOH/NH ₄ OH (97/3) to obtain (R)-quinuclidin-3-yl 2- amino-2-phenylacetate dihydrochloride (55.0 mg; 65.1% yield).

EXAMPLE 9

Preparation of (3R)-3-(2-(benzyloxycarbonylamino)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane bromide (C31)

Scheme 10

To a solution of (R)-quinuclidin-3-yl 2-(benzyloxycarbonylamino)-2- phenylacetate (C29) (100 mg, 0.25 mmol) in EtOAc (5 ml), 2-bromo- l-phenylethanone (55.5 mg, 0.28 mmol) was added and the reaction was stirred at RT for 15h. The solvent was evaporated and the resulting crude was purified by silica gel flash chromatography (DCM/MeOH=93/7) to obtain (3R)-3-(2-(benzyloxycarbonylamino)-2-phenylacetoxy)- l-(2-oxo-2- phenylethyl)- l-azoniabicyclo[2.2.2]octane bromide (127.5 mg; 85% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 8.22 - 8.52 (m, 1 H), 7.90 - 8.07 (m, 2 H), 7.69 - 7.83 (m, 1 H), 7.54 - 7.69 (m, 2 H), 7. 19 - 7.54 (m, 10 H), 5.39 and 5.40 (d, 1 H), 4.99 - 5.29 (m, 5 H), 3.93 - 4.23 (m, 1 H), 3.39 - 3.78 (m, 5 H), 2. 10 - 2.23 and 2.30 - 2.44 (m, 1 H), 1.40 - 2. 12 (m, 4 H);

LC-MS (ESI POS): 513.25 (M+).

EXAMPLE 10

Preparation of (R)-((R)-quinuclidin-3-yl)

2-(benzyloxycarbonylamino)-2-phenylacetate (Diastereoisomer 1 of C31)

Diastereoisomer 1 of C29 Diastereoisomer 1 of C31

Scheme 11

Preparation of (R)-((R)-quinuclidin-3-yl)

2-(benzyloxycarbonylamino)-2-phenylacetate (Diastereoisomer 1 of C29):

(R)-quinuclidin-3-ol (214 mg, 1.68 mmol), N,N'-methanediylidenedicyclohexanamine (347 mg, 1.68 mmol) and lH-benzo[d] [ l,2,3]triazol- l-ol (227 mg, 1.68 mmol) were added to a solution of (R)-2-(benzyloxycarbonylamino)-2-phenylacetic acid (400 mg, 1.40 mmol) in THF (15 ml). The reaction was stirred at RT for 15h then the solvent was evaporated. DCM was added and the insoluble solid was filtered off. The organic phase was washed twice with Na ₂ CO ₃ and brine, dried over Na ₂ SO ₄ and evaporated. The crude was purified by silica gel flash chromatography (DCM/MeOH=9/l) to obtain (R)-((R)-quinuclidin-3-yl)

2-(benzyloxycarbonylamino)-2-phenylacetate (63 mg; 1 1.4% yield). ((R)-Quinuclidin-3-yl 2-(benzyloxycarbonylamino)-2-phenylacetate is also collected (300 mg; 54.2% yield)).

1H NMR (300 MHz, DMSO-<¾) δ ppm 8.26 (d, 1 H), 7.14 - 7.58 (m, 10 H), 5.28 (d, 1 H), 5. 10 (d, 1 H), 5.05 (d, 1 H), 4.59 - 4.79 (m, 1 H), 3.01 (dd, 1 H), 2.54 - 2.70 (m, 3 H), 2.32 - 2.45 (m, 1 H), 2.22 (d, 1 H), 1.82 - 1.99 (m, 1 H), 1.35 - 1.68 (m, 3 H), 1.16 - 1.35 (m, 1 H).

Preparation of (R)-3-((R)-2-(benzyloxycarbonylamino)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo [2.2.2]octane bromide (Diastereoisomer 1 of C31)

To a solution of (R)-((R)-quinuclidin-3-yl) 2-(benzyloxy- carbonylamino)-2-phenylacetate (40 mg, 0.10 mmol) in EtOAc (3 ml), 2-bromo- l-phenylethanone (22.2 mg, 0. 1 1 mmol) was added and the reaction was stirred at RT for 15h then the solvent was evaporated. The crude was purified by silica gel flash chromatography (DCM/MeOH =93/7) to obtain (R)-3-((R)-2-(benzyloxycarbonylamino)-2-phenylacetoxy)- 1 -(2-0X0-2- phenylethyl)- l-azoniabicyclo[2.2.2]octane bromide (45.0 mg; 74.8% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 8.37 (d, 1 H), 7.87 - 8.04 (m, 2 H), 7.69 - 7.82 (m, 1 H), 7.56 - 7.69 (m, 2 H), 7.18 - 7.54 (m, 10 H), 5.38 (d, 1 H), 5. 17 - 5.26 (m, 1 H), 5. 15 (s, 2 H), 5.09 (s, 2 H), 3.95 - 4.21 (m, 1 H), 3.42 - 3.75 (m, 5 H), 2.3 1 - 2.44 (m, 1 H), 1.39 - 2. 12 (m, 4 H);

LC-MS (ESI POS): 513. 17 (M+).

EXAMPLE 11

Preparation of (3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2- (vinyloxycarbonylamino)acetoxy)-l-azoniabicyclo [2.2.2] octane bromide (C33)

Scheme 12

Preparation of (R)-quinuclidin-3-yl 2-phenyl-2- (vinyloxycarbonylamino)acetate (C32)

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (110 mg, 0.33 mmol) in DCM (5 ml), triethylamine (138 μΐ, 0.99 mmol) and vinyl carbonochloridate (36.2 μΐ, 0.40 mmol) were added. The reaction was stirred at RT for 4h, then the solvent was evaporated. The residue was taken up with EtOAc and washed with water and brine, dried over Na ₂ SO ₄ and evaporated to dryness to obtain (R)-quinuclidin-3-yl 2-phenyl-2-(vinyloxycarbonylamino)acetate (70 mg; 64% yield). The product was used in the next step without any further purification.

Preparation of (3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2-

(vinyloxycarbonylamino)acetoxy)-l-azoniabicyclo [2.2.2] octane bromide (C33)

To a solution of (R)-quinuclidin-3-yl 2-phenyl-2- (vinyloxycarbonylamino)acetate (C32) (70.0 mg, 0.21 mmol) in EtOAc (2 ml) and acetonitrile (0.5 ml), 2-bromo- l-phenylethanone (46.4 mg, 0.23 mmol) was added. The reaction was stirred at RT for 15h and then the solvent was evaporated. The resulting solid was triturated with z-Pr ₂ O/EtOAc (1/1) to obtain (3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2-(vinyloxycarbonyl amino)- acetoxy)-l-azoniabicyclo[2.2.2]octane bromide (82.4 mg; 73% yield).

1H NMR (300 MHz, DMSO-d6) δ ppm 8.76 and 8.78 (d, 1 H),

7.90 - 8.08 (m, 2 H), 7.71 - 7.80 (m, 1 H), 7.56 - 7.67 (m, 2 H), 7.30 - 7.54 (m, 5 H), 7.07 - 7.20 (m, 1 H), 5.41 and 5.42 (d, 1 H), 5.20 - 5.30 (m, 1 H), 5.15 and 5.17 (s, 2 H), 4.78 (dd, 1 H), 4.53 (dd, 1 H), 3.97 - 4.22 (m, 1 H), 3.43 - 3.83 (m, 5 H), 2.15 - 2.24 and 2.35 - 2.44 (m, 1 H), 1.50 - 2.14 (m, 4 H);

LC-MS (ESI POS): 449.27 (M+). EXAMPLE 12

Preparation of (3R)-3-(2-(ethoxycarbonylamino)-2-phenylacetoxy)- l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide (C35)

Scheme 13

Preparation of (R)-quinuclidin-3-yl 2-(ethoxycarbonylamino)-2- phenylacetate (C34)

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (1 10 mg, 0.33 mmol) in DCM (5 ml), triethylamine (138 μΐ, 0.99 mmol) and ethyl carbonochloridate (38 μΐ, 0.40 mmol) were added. The reaction was stirred at RT for 4h and then solvent was evaporated. EtOAc was added and the organic phase was washed with water and brine, dried over Na ₂ SO ₄ and evaporated to obtain (R)-quinuclidin-3-yl 2-(ethoxycarbonylamino)-2-phenylacetate (60 mg; 55% yield). The product was used as such in the next step.

Preparation of (3R)-3-(2-(ethoxycarbonylamino)-2-phenylacetoxy)- l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide (C35)

To a solution of (R)-quinuclidin-3-yl 2-(ethoxycarbonylamino)-2- phenylacetate (C34) (60.0 mg, 0.18 mmol) in EtOAc (2 ml) and acetonitrile (0.5 ml), 2-bromo- l-phenylethanone (39.5 mg, 0.20 mmol) was added. The reaction was stirred at RT for 15h and the solvent was evaporated. The resulting solid was triturated in z-Pr ₂ O/EtOAc (1/1) to obtain (3R)-3-(2- (ethoxycarbonylamino)-2-phenylacetoxy)-l-(2-oxo-2-phenylethy l)-l- azoniabicyclo[2.2.2]octane bromide (76 mg; 80% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 8.1 1 - 8.29 (m, 1 H), 7.91 - 8.03 (m, 2 H), 7.70 - 7.81 (m, 1 H), 7.56 - 7.67 (m, 2 H), 7.29 - 7.54 (m, 5 H), 5.35 and 5.36 (d, 1 H), 5.20 - 5.26 (m, 1 H), 5.16 and 5.18 (s, 2 H), 4.09 - 4.22 (m, 1 H), 4.06 (q, 2 H), 3.43 - 3.84 (m, 5 H), 2.15 - 2.24 (m, 1 H), 1.52 - 2.14 and 2.32 - 2.42 (m, 4 H), 1.19 (t, 3 H);

LC-MS (ESI POS): 451.27 (M+).

EXAMPLE 13

Preparation of (3R)-3-(2-((2-methoxyethoxy)carbonylamino)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane bromide (C37)

Scheme 14

Preparation of (R)-quinuclidin-3-yl 2-((2-methoxyethoxy)- carbonylamino)-2-phenylacetate (C36):

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (150 mg, 0.45 mmol) in DCM (5 ml), triethylamine (188 μΐ, 1.35 mmol) and 2-methoxyethyl carbonochloridate (63 μΐ, 0.54 mmol) were added. The reaction was stirred at RT for 4h and then the solvent was evaporated. The residue was taken up with EtOAc and the organic phase was washed with water and brine, dried over Na ₂ SO ₄ and evaporated to dryness, obtaining (R)-quinuclidin-3-yl 2-((2-methoxyethoxy)carbonylamino)- 2-phenylacetate (73.0 mg; 45% yield).

Preparation of (3R)-3-(2-((2-methoxyethoxy)carbonylamino)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo [2.2.2]octane bromide (C37)

To a solution of (R)-quinuclidin-3-yl 2-((2-methoxyethoxy)- carbonylamino)-2-phenylacetate (C36) (73.0 mg, 0.20 mmol) in EtOAc (3 ml) and acetonitrile (1 ml), 2-bromo- l-phenylethanone (44. 1 mg, 0.22 mmol) was added. The reaction was stirred at RT for 15h then the solvent was evaporated and the resulting crude was purified by flash chromatography (DCM/MeOH =95/5 to 93/7). The product was triturated with z-Pr ₂ O to obtain (3R)-3-(2-((2- methoxyethoxy)carbonylamino)-2-phenylacetoxy)- l-(2-oxo-2-phenylethyl)- l- azoniabicyclo[2.2.2]octane bromide (61.3 mg; 54% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 8.23 - 8.44 (m, 1 H) 7.89 - 8.08 (m, 2 H) 7.69 - 7.83 (m, 1 H) 7.54 - 7.68 (m, 2 H) 7.29 - 7.52 (m, 5 H) 5.36 (dd, 1 H) 5.08 - 5.28 (m, 3 H) 4.05 - 4.21 (m, 3 H) 3.44 - 3.80 (m, 7 H) 3.26 (s, 3 H) 2.32 - 2.43 (m, 1 H) 1.88 - 2. 12 (m, 3 H) 1.69 - 1.88 (m, 1 H);

LC-MS (ESI POS): 481. 18 (M+).

EXAMPLE 14

Preparation of (3R)-3-(2-(cyclohexyloxycarbonylamino)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane bromide (C39)

Scheme 15 Preparation of (R)-quinuclidin-3-yl 2-

(cyclohexyloxycarbonylamino)-2-phenylacetate (C38)

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (150 mg, 0.45 mmol) in DCM (5 ml), triethylamine (188 μΐ, 1.35 mmol) and cyclohexyl carbonochloridate (78 μΐ, 0.54 mmol) were added. The reaction was stirred at RT for 4h and then the solvent was evaporated.

The residue was taken up with EtOAc and washed with water and brine, dried over Na ₂ SO ₄ and evaporated to dryness to obtain (R)-quinuclidin-3-yl 2-(cyclohexyloxycarbonylamino)-2-phenylacetate (105 mg; 60% yield).

Preparation of (3R)-3-(2-(cyclohexyloxycarbonylamino)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane bromide (C39)

To a solution of (R)-quinuclidin-3-yl 2-(cyclohexyloxycarbonylamino)- 2-phenylacetate (C38) (105 mg, 0.27 mmol) in EtOAc (3 ml), 2-bromo- l- phenylethanone (59.5 mg, 0.30 mmol) was added. The reaction was stirred at RT for 15h then the solvent was evaporated. The crude was purified by flash chromatography (DCM/MeOH = 95/5 to 93/7) to obtain (3R)-3-(2- (cyclohexyloxycarbonylamino)-2-phenylacetoxy)-l-(2-oxo-2-phe nylethyl)- l- azoniabicyclo[2.2.2]octane bromide (83.8 mg; 53% yield).

1H NMR (300 MHz, DMSO-< ) δ ppm 8.13 (m, 1 H) 7.90 - 8.04 (m, 2 H) 7.69 - 7.84 (m, 1 H) 7.61 (td, 2 H) 7.25 - 7.53 (m, 5 H) 5.35 (dd, 1 H) 5.05 - 5.28 (m, 3 H) 4.41 - 4.67 (m, 1 H) 3.91 - 4.23 (m, 1 H) 3.49 - 3.76 (m, 5 H) 2.12 - 2.24 (m, 1 H) 1.89 - 2.12 (m, 3 H) 1.57 - 1.89 (m, 5 H) 1.1 1 - 1.56 (m, 6 H);

LC-MS (ESI POS): 505.19 (M+).

EXAMPLE 15

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2-p- tolylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2] octane bromide (C42)

C42

Scheme 16

Preparation of 2-(tert-butoxycarbonylamino)-2-p-tolylacetic acid

(140):

To a suspension of 2-amino-2-p-tolylacetic acid (1.00 g, 6.05 mmol) in THF (30 ml) and water (30 ml), 2N sodium hydroxide (30.3 ml, 60.5 mmol) and di-tert-butyl dicarbonate (2.64 g, 12.1 mmol) were added. The reaction was stirred at RT for 15h then THF was evaporated. The remaining aqueous phase was cooled and acidified with 37% HC1 until pH 1. The desired compound was extracted with EtOAc and the organic phase was washed with brine, dried over Na ₂ SO ₄ and evaporated to obtain 2-(tert-butoxycarbonylamino)-2-p-tolylacetic acid (1.29 g; 80% yield). Preparation of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)- 2-p-tolylacetate (C41)

To a solution of 2-(tert-butoxycarbonylamino)-2-p-tolylacetic acid (140) (1.29 g, 4.86 mmol) in THF (70 ml), Ν,Ν'- methanediylidenedicyclohexanamine (1.20 g, 5.83 mmol), 1H- benzo[d][l,2,3]triazol-l-ol (0.79 g, 5.83 mmol) and (R)-quinuclidin-3-ol (0.74 g, 5.83 mmol) were added. The reaction was stirred at RT for 15h and then solvent was evaporated. The residue was taken up with DCM, the insoluble solid was filtered off and the organic solution was washed twice with aq. Na ₂ CO ₃ and then brine, dried over Na ₂ SO ₄ and evaporated to dryness. The crude was purified by flash chromatography (EtO Ac/Me OH= 8/2 to 7/3) to obtain (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-p-tolylacetate (1.03 g; 57% yield).

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2-p- tolylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2] octane bromide (C42):

To a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2- p-tolylacetate (C41) (100 mg, 0.27 mmol) in EtOAc (3 ml), 2-bromo-l- phenylethanone (58.5 mg, 0.29 mmol) was added and the reaction was stirred at RT for 36h. The solvent was evaporated and the residue was first triturated with 7-Pr ₂ O and then purified by flash chromatography (DCM/MeOH =95/5 to 9/1) to obtain (3R)-3-(2-(tert-butoxycarbonylamino)-2-p-tolylacetoxy)-l-(2- oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide (94.4 mg; 62% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 7.92 - 8.06 (m, 2 H), 7.70 - 7.86

(m, 2 H), 7.54 - 7.68 (m, 2 H), 7.27 - 7.42 (m, 2 H), 7.12 - 7.25 (m, 2 H), 4.91 - 5.36 (m, 3 H), 3.92 - 4.21 (m, 1 H), 3.44 - 3.85 (m, 5 H), 2.34 - 2.40 (m, 1 H), 2.30 (s, 3 H), 1.53 - 2.22 (m, 4 H), 1.41 (s, 9 H); LC-MS (ESI POS): 493.61 (M+).

EXAMPLE 16

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2-(4- methoxyphenyl)acetoxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane bromide (C45)

C45

Scheme 17

Preparation of 2-(tert-butoxycarbonylamino)-2-(4- methoxyphenyl)acetic acid (143)

To a suspension of 2-amino-2-(4-methoxyphenyl)acetic acid (360 mg, 1.99 mmol) in THF (30 ml) and water (30 ml), 2N sodium hydroxide (20 ml, 40.0 mmol) and di-tert-butyl dicarbonate (867 mg, 3.97 mmol) were added. The reaction was stirred at RT for 15h. THF was evaporated, and the remaining aqueous phase was cooled and acidified with 37% HCl until pH 1. The desired compound was extracted with EtOAc and the organic phase was washed with brine, dried over Na ₂ SO ₄ and evaporated to afford 2-(tert- butoxycarbonylamino)-2-(4-methoxyphenyl)acetic acid (430 mg; 77% yield).

Preparation of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)- 2-(4-methoxyphenyl)acetate (C44)

To a solution of 2-(tert-butoxycarbonylamino)-2-(4- methoxyphenyl)acetic acid (143) (1.39 g, 4.94 mmol) in dry THF (60 ml), Ν,Ν'-methanediylidenedicyclohexanamine (1.22 g, 5.94 mmol), lH-benzo[d][l,2,3]triazol-l-ol (802 mg, 5.94 mmol) and (R)-quinuclidin-3-ol (755 mg, 5.94 mmol) were added. The reaction was stirred at RT for 15h and the solvent was evaporated. The residue was taken up with DCM, the insoluble solid was filtered off and the organic solution was washed twice with aq. Na ₂ CO ₃ and then brine, dried over Na ₂ SO ₄ and evaporated. The crude was purified by flash chromatography (DCM/MeOH= 95/5 to 93/7) to obtain (R)-quinuclidin-3 -yl 2-(tert-butoxycarbonylamino)-2-(4- methoxyphenyl)acetate (630 mg; 33% yield).

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2-(4- methoxyphenyl)acetoxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane bromide (C45)

2-Bromo-l-phenylethanone (39.3 mg, 0.20 mmol) is added to a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(4- methoxyphenyl)acetate (C44) (70.0 mg, 0.18 mmol) in EtOAc (2 ml) and acetonitrile (2 ml). The reaction was stirred at RT for 15h then the solvent was evaporated and the resulting colorless oil was triturated first with 7-Pr ₂ O/EtOAc (10/1) and then with z-Pr ₂ O to obtain (3R)-3-(2-(tert- butoxycarbonylamino)-2-(4-methoxyphenyl)acetoxy)- 1 -(2-oxo-2- phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide (60.9 mg; 58% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 7.88 - 8.05 (m, 2 H), 7.71 - 7.84 (m, 2 H), 7.54 - 7.66 (m, 2 H), 7.27 - 7.44 (m, 2 H), 6.80 - 7.02 (m, 2 H), 5.18 - 5.26 (m, 2 H), 5.16 (s, 2 H), 4.02 - 4.20 (m, 1 H), 3.76 (s, 3 H), 3.47 - 3.72 (m, 5 H), 2.33 - 2.44 (m, 1 H), 1.78 - 2.17 (m, 4 H), 1.41 (s, 9 H);

LC-MS (ESI POS): 509.15 (M+).

EXAMPLE 17

Preparation of (3R)-3-(2-(tert-butoxycarbonylamin chlorophenyl)acetoxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane bromide (C48)

C48

Scheme 18

Preparation of 2-(tert-butoxycarbonylamino)-2-(4- chlorophenyl)acetic acid (146):

To a suspension of 2-amino-2-(4-chlorophenyl)acetic acid (1.50 g, 8.08 mmol) in THF (30 ml) and water (30 ml), 2N sodium hydroxide (40.4 ml, 81.0 mmol) and di-tert-butyl dicarbonate (3.53 g, 16.2 mmol) were added. The reaction was stirred at RT for 15h. THF was evaporated, and the remaining aqueous phase was cooled and acidified with 37% HCl until pH 1. The desired compound was extracted with EtOAc and the organic phase was washed with brine, dried over Na ₂ SO ₄ and evaporated to afford 2-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)acetic acid (2. 17 g; 94% yield).

Preparation of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)- 2-(4-chlorophenyl)acetate (C47):

To a solution of 2-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)acetic acid (146) (2. 17 g, 7.58 mmol) in dry THF (70 ml), Ν,Ν'- methanediylidenedicyclohexanamine (1.88 g, 9.10 mmol), 1H- benzo[d][ l,2,3]triazol- l-ol (1.23 g, 9. 10 mmol) and (R)-quinuclidin-3-ol (1. 16 g, 9. 10 mmol) were added. The reaction was stirred at RT for 15h and the solvent was evaporated. The residue was taken up with DCM, the insoluble solid was filtered off and the organic solution was washed twice with aq. Na ₂ CO ₃ and then brine, dried over Na ₂ SO ₄ and evaporated. The crude was purified by flash chromatography (EtO Ac/Me OH= 8/2 to 85/15) to obtain (R)-quinuclidin-3 -yl 2-(tert-butoxycarbonylamino)-2-(4-chlorophenyl)acetate (1.33 g; 44% yield).

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2-(4- chlorophenyl)acetoxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo [2.2.2]octane bromide (C48):

2-Bromo- l-phenylethanone (55.4 mg, 0.28 mmol) was added to a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(4- chlorophenyl)acetate (C47) (100 mg, 0.25 mmol) in EtOAc (3 ml). The reaction was stirred at RT for 36h then a second portion of 2-bromo- l- phenylethanone (50.4 mg, 0.25 mmol) was added and the reaction was stirred at RT for additional 48h. The organic phase was washed with aq. Na ₂ CO ₃ , dried over Na ₂ SO ₄ and evaporated. The crude was dissolved in acetonitrile (3 ml) and 2-bromo- l-phenylethanone (60.0 mg, 0.30 mmol) was added. The reaction was heated under microwave irradiation at 100°C for 45 minutes. The solvent was evaporated and the crude was purified by flash chromatography (DCM/MeOH=95/5 to 9/1) to obtain (3R)-3-(2-(tert-butoxycarbonylamino)-2- (4-chlorophenyl)acetoxy)-l-(2-oxo-2-phenylethyl)- l- azoniabicyclo[2.2.2]octane bromide (62.5 mg; 42% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 7.86 - 8.03 (m, 3 H), 7.69 - 7.82 (m, 1 H), 7.55 - 7.67 (m, 2 H), 7.37 - 7.55 (m, 4 H), 5.34 (d, 1 H), 5.16 - 5.25 (m, 1 H), 5.05 - 5.16 (m, 1 H), 3.96 - 4.16 (m, 1 H), 3.43 - 3.81 (m, 5 H), 2.32 - 2.42 (m, 1 H), 1.79 - 2.15 (m, 4 H), 1.41 (s, 9 H);

LC-MS (ESI POS): 513.16 (M+).

EXAMPLE 18

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2-(4- fluorophenyl)acetoxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane bromide (C51)

C51

Scheme 19

Preparation of 2-(tert-butoxycarbonylamino)-2-(4- fluorophenyl)acetic acid (149)

To a solution of 2-amino-2-(4-fluorophenyl)acetic acid (1.00 g, 5.91 mmol) in acetone (50 ml) and 2N sodium hydroxide (50 ml, 100 mmol), di-tert-butyl dicarbonate (1.29 g, 5.91 mmol) was added and the reaction was stirred at RT for 3h. Acetone was evaporated, the aqueous phase was acidified with 37% HCl until pH 1. The product was extracted with EtOAc and the organic phase was washed with water and brine, dried over Na ₂ SO ₄ and evaporated. The resulting oil was triturated with petroleum ether to give 2-(tert-butoxycarbonylamino)-2-(4-fluorophenyl)acetic acid (725 mg; 45% yield).

Preparation of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)- 2-(4-fluorophenyl)acetate (C50)

To a solution of 2-(tert-butoxycarbonylamino)-2-(4-fluorophenyl)acetic acid (149) (725 mg, 2.69 mmol) in THF (20 ml), Ν,Ν'- methanediylidenedicyclohexanamine (667 mg, 3.23 mmol), lH-benzo[d][l,2,3]triazol-l-ol (437 mg, 3.23 mmol) and (R)-quinuclidin-3-ol (411 mg, 3.23 mmol) were added. The reaction was stirred at RT for 15h and the solvent was evaporated. The residue was taken up with DCM, the insoluble solid was filtered off and the organic solution was washed twice with aq. Na ₂ CO ₃ and then brine, dried over Na ₂ SO ₄ and evaporated. The crude was purified by flash chromatography (DCM/MeOH= 95/5 to 93/7) to obtain (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(4-fluorophenyl)acetate (410 mg; 40% yield).

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2-(4- fluorophenyl)acetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicycl o [2.2.2]- octane bromide (C51)

2-Bromo-l-phenylethanone (34.7 mg, 0.17 mmol) is added to a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(4-fluorophenyl)- acetate (C50) (60.0 mg, 0. 16 mmol) in EtOAc (2 ml) and acetonitrile (2 ml). The reaction was stirred at RT for 15h then the solvent was evaporated. The resulting residue was triturated with z-Pr ₂ O/EtOAc ( 10/1) and then z-Pr ₂ O to obtain (3R)-3-(2-(tert-butoxycarbonylamino)-2-(4-fluorophenyl)aceto xy)- l- (2-oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane bromide (46.1 mg; 50% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 7.93 - 8.02 (m, 2 H), 7.91 (d, 1 H), 7.69 - 7.80 (m, 1 H), 7.57 - 7.67 (m, 2 H), 7.47 - 7.57 (m, 2 H), 7.17 - 7.3 1 (m, 2 H), 5.33 (d, 1 H), 5.17 - 5.26 (m, 1 H), 5. 15 (s, 2 H), 4.07 (ddd, 1 H), 3.44 - 3.79 (m, 5 H), 2.37 (br. s., 1 H), 1.74 - 2.23 (m, 4 H), 1.41 (s, 9 H);

LC-MS (ESI POS): 497. 14 (M+).

EXAMPLE 19

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2-(3- fluorophenyl)acetoxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane bromide (C54)

C54

Scheme 20 Preparation of 2-(tert-butoxycarbonylamino)-2-(3- fluorophenyl)acetic acid (152):

To a suspension of 2-amino-2-(3-fluorophenyl)acetic acid (1.00 g, 5.91 mmol) in THF (30 ml) and water (30 ml), 2N sodium hydroxide (29.6 ml, 59.1 mmol) and di-tert-butyl dicarbonate (2.58 g, 1 1.8 mmol) were added and the reaction was stirred at RT for 15h. THF was evaporated, the aqueous phase was cooled and acidified with 37% HCl until pH 1. The desired compound was extracted with EtOAc and the organic phase was washed with brine, dried over Na ₂ SO ₄ and evaporated to obtain 2-(tert-butoxycarbonylamino)-2-(3- fluorophenyl)acetic acid (1.10 g; 69% yield).

Preparation of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)- 2-(3-fluorophenyl)acetate (C53):

To a solution of 2-(tert-butoxycarbonylamino)-2-(3-fluorophenyl)acetic acid (152) (1.10 g, 4.09 mmol) in THF (50 ml), Ν,Ν'- methanediylidenedicyclohexanamine (1.01 g, 4.90 mmol), 1H- benzo[d][l,2,3]triazol-l-ol (0.66 g, 4.90 mmol) and (R)-quinuclidin-3-ol (0.62 g, 4.90 mmol) were added. The reaction was stirred at RT for 15h and the solvent was evaporated. The residue was taken up with DCM, the insoluble solid was filtered off and the organic solution was washed twice with aq. Na ₂ CO ₃ and then brine, dried over Na ₂ SO ₄ and evaporated to obtain (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(3-fluorophenyl)acetate (1.55 g; quantitative yield).

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2-(3- fluorophenyl)acetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicycl o [2.2.2]- octane bromide (C54)

2-Bromo-l-phenylethanone (63.1 mg, 0.32 mmol) was added to a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(3- fluorophenyl)acetate (C53) (100 mg, 0.26 mmol) in EtOAc (3 ml) and acetonitrile (3 ml). The reaction was stirred at RT for 15h then the solvent was evaporated. The crude was purified by flash chromatography (DCM/MeOH=

94/6) to obtain (3R)-3-(2-(tert-butoxycarbonylamino)-2-(3- fluorophenyl)acetoxy)- l-(2-oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane bromide (70.7 mg; 46% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 7.87 - 8.07 (m, 3 H), 7.70 - 7.83 (m, 1 H), 7.55 - 7.69 (m, 2 H), 7.38 - 7.50 (m, 1 H), 7.27 - 7.38 (m, 2 H), 7.08 - 7.27 (m, 1 H), 5.32 - 5.49 (m, 1 H), 5. 19 - 5.27 (m, 1 H), 5.09 - 5. 19 (m, 1 H), 3.93 - 4.23 (m, 1 H), 3.42 - 3.90 (m, 5 H), 2. 15 - 2.26 and 2.32 - 2.42 (m, 1 H), 1.59 - 2. 13 (m, 4 H), 1.41 (s, 9 H);

LC-MS (ESI POS): 497.22 (M+).

EXAMPLE 20

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2-(2- fluorophenyl)acetoxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane bromide (C57)

Scheme 21 Preparation of 2-(tert-butoxycarbonylamino)-2-(2- fluorophenyl)acetic acid (155):

To a suspension of 2-amino-2-(2-fluorophenyl)acetic acid (1.00 g, 5.91 mmol) in THF (30 ml) and water (30 ml), 2N sodium hydroxide (29.6 ml, 59.1 mmol) and di-tert-butyl dicarbonate (2.58 g, 11.8 mmol) were added. The reaction was stirred at RT for 15h. THF was evaporated, the aqueous phase was cooled to 0°C and acidified with 37% HC1 until pH 1. The desired compound was extracted with EtOAc and the organic phase was washed with brine, dried over Na ₂ SO ₄ and evaporated to obtain 2-(tert- butoxycarbonylamino)-2-(2-fluorophenyl)acetic acid (1.11 g; 70% yield).

Preparation of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)- 2-(2-fluorophenyl)acetate (C56)

To a solution of 2-(tert-butoxycarbonylamino)-2-(2-fluorophenyl)acetic acid (155) (1.11 g, 4.12 mmol) in THF (50 ml), Ν,Ν'- methanediylidenedicyclohexanamine (1.02 g, 4.95 mmol), 1H- benzo[d][l,2,3]triazol-l-ol (0.67 g, 4.95 mmol) and (R)-quinuclidin-3-ol (0.63 g, 4.95 mmol) were added. The reaction was stirred at RT for 15h and the solvent was evaporated. The residue was taken up with DCM, the insoluble solid was filtered off and the organic solution was washed twice with aq. Na ₂ CO ₃ and then brine, dried over Na ₂ SO ₄ and evaporated to obtain (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(2-fluorophenyl)acetate (1.56 g; quantitative yield).

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2-(2- fluorophenyl)acetoxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane bromide (C57)

2-Bromo-l-phenylethanone (57.9 mg, 0.29 mmol) was added to a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(2- fluorophenyl)acetate (C56) (100 mg, 0.26 mmol) in EtOAc (3 ml) and acetonitrile (3 ml). The reaction was stirred at RT for 15h then the solvent was evaporated. The crude was purified by flash chromatography (DCM/MeOH= 94/6) to obtain (3R)-3-(2-(tert-butoxycarbonylamino)-2-(2- fluorophenyl)acetoxy)- l-(2-oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane bromide (95. 1 mg; 62% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 7.90 - 8. 10 (m, 3 H), 7.70 - 7.81 (m, 1 H), 7.61 (td, 2 H), 7.47 - 7.56 (m, 1 H), 7.37 - 7.47 (m, 1 H), 7. 15 - 7.30 (m, 2 H), 5.49 - 5.68 (m, 1 H), 5.20 - 5.37 (m, 1 H), 5.04 - 5.20 (m, 1 H), 3.99 - 4.28 (m, 1 H), 3.39 - 3.89 (m, 5 H), 2.32 - 2.43 (m, 1 H), 1.48 - 2. 15 and 2. 15 - 2.25 (m, 4 H), 1.41 (s, 9 H);

LC-MS (ESI POS): 497. 18 (M+).

EXAMPLE 21

Preparation of 3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)- 8-methyl-8-(2-oxo-2-phenylethyl)-8-azoniabicyclo[3.2.1]octan e bromide (C59)

13 C58 C59

Scheme 22

Preparation of 8-methyl-8-azabicyclo[3.2.1]octan-3-yl 2-(tert- butoxycarbonylamino)-2-phenylacetate (C58)

To a solution of 2-(tert-butoxycarbonylamino)-2-phenylacetic acid (400 mg, 1.59 mmol) in THF (20 ml), Ν,Ν'- methanediylidenedicyclohexanamine (394 mg, 1.91 mmol), 1H- benzo[d][l,2,3]triazol-l-ol (258 mg, 1.91 mmol) and 8-methyl-8- azabicyclo[3.2.1]octan-3-ol (270 mg, 1.91 mmol) were added. The reaction was stirred at RT for 15h then the solvent was evaporated. The residue was taken up with DCM, the insoluble was filtered off and the clear solution was washed twice with aq. Na ₂ CO ₃ and brine, dried over Na ₂ SO ₄ and evaporated to give 8-methyl-8-azabicyclo[3.2.1]octan-3-yl 2-(tert-butoxycarbonylamino)- 2-phenylacetate (460 mg; 77% yield). The product was used in the next step without any further purification.

Preparation of 3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)- 8-methyl-8-(2-oxo-2-phenylethyl)-8-azoniabicyclo [3.2.1] octane bromide (C59)

To a solution of 8-methyl-8-azabicyclo[3.2.1]octan-3-yl 2-(tert- butoxycarbonylamino)-2-phenylacetate (C58) (230 mg, 0.61 mmol) in DMF (15 ml) and acetonitrile (5 ml), 2-bromo- l-phenylethanone (134 mg, 0.68 mmol) was added. The reaction was stirred at RT for 15h and then the solvent was evaporated. The crude was first triturated with z-Pr ₂ O and then purified by flash chromatography (DCM/MeOH=94/6) to obtain 3-(2-(tert- butoxycarbonylamino)-2-phenylacetoxy)-8-methyl-8-(2-oxo-2-ph enylethyl)-8- azoniabicyclo[3.2.1]octane bromide (196 mg; 56% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 7.96 - 8.05 (m, 2 H) 7.88 (d, 1

H) 7.67 - 7.79 (m, 1 H) 7.52 - 7.66 (m, 2 H) 7.29 - 7.51 (m, 5 H) 5.29 (d, 1 H) 5.09 (s, 2 H) 5.00 - 5.07 (m, 1 H) 4.27 - 4.42 (m, 1 H) 4.15 - 4.27 (m, 1 H) 3.26 (s, 3 H) 2.57 - 2.76 (m, 2 H) 2.04 - 2.42 (m, 3 H) 1.87 - 2.02 (m, 1 H) 1.48 - 1.80 (m, 2 H) 1.40 (s, 9 H);

LC-MS (ESI POS): 493.30 (M+). EXAMPLE 22

Preparation of 3-(2-(tert-butoxycarbonylamino)-2-phenylacetoxy)- 8,8-dimethyl-8-azoniabicyclo[3.2.1]octane iodide (C60)

C58 C60

Scheme 23

To a solution of 8-methyl-8-azabicyclo[3.2.1]octan-3-yl 2-(tert- butoxycarbonylamino)-2-phenylacetate (C58) (230 mg, 0.61 mmol) in DMF (15 ml), iodomethane (42.1 μΐ, 0.68 mmol) was added. The reaction was stirred at RT for 15h and then the solvent was evaporated. The crude was triturated with z-Pr ₂ O and then purified by flash chromatography (DCM/MeOH=95/5 to 9/1) to obtain 3-(2-(tert-butoxycarbonylamino)-2- phenylacetoxy)-8,8-dimethyl-8-azoniabicyclo[3.2.1]octane iodide (88.2 mg: 28% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 7.85 (d, 1 H) 7.22 - 7.62 (m, 5 H) 5.26 (d, 1 H) 4.98 (t, 1 H) 3.76 - 3.87 (m, 1 H) 3.65 - 3.76 (m, 1 H) 3.06 (s, 3 H) 2.96 (s, 3 H) 2.57 - 2.65 (m, 2 H) 1.95 - 2.23 (m, 3 H) 1.87 (d, 1 H) 1.46 - 1.67 (m, 2 H) 1.40 (s, 9 H);

LC-MS (ESI POS): 389.25 (M+). EXAMPLE 23

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2- phenylacetoxy)-l-methyl-l-(2-oxo-2-phenylethyl)pyrrolidinium bromide (C62)

Scheme 24

Preparation of (R)-l-methylpyrrolidin-3-yl 2-(tert- butoxycarbonylamino)-2-phenylacetate (C61)

To a solution of 2-(tert-butoxycarbonylamino)-2-phenylacetic acid (13)

(400 mg, 1.59 mmol) in THF (20 ml), (R)-l-methylpyrrolidin-3-ol (193 mg, 1.91 mmol), N,N'-methanediylidenedicyclohexanamine (394 mg, 1.91 mmol) and lH-benzo[d][ l,2,3]triazol-l-ol (258 mg, 1.91 mmol) were added. The reaction was stirred at RT for 15h then the solvent was evaporated. The residue was taken up with DCM, the insoluble was filtered off and the clear solution was washed twice with aq. Na ₂ CO ₃ and brine, dried over Na ₂ SO ₄ and evaporated to obtain (R)-l-methylpyrrolidin-3-yl

2-(tert-butoxycarbonylamino)-2-phenylacetate (340 mg; 64% yield).

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2- phenylacetoxy)-l-methyl-l-(2-oxo-2-phenylethyl)pyrrolidinium bromide (C62)

To a solution of (R)- l-methylpyrrolidin-3-yl

2-(tert-butoxycarbonylamino)-2-phenylacetate (C61) (170 mg, 0.51 mmol) in DMF (5 ml) and acetonitrile (3 ml), 2-bromo-l-phenylethanone (111 mg, 0.56 mmol) was added. The reaction was stirred at RT for 15h and the solvent was evaporated. The crude was purified by flash chromatography (DCM/MeOH=9/l) to obtain (3R)-3-(2-(tert-butoxycarbonylamino)-2- phenylacetoxy)- 1 -methyl- l-(2-oxo-2-phenylethyl)pyrrolidinium bromide (152 mg; 56% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 7.69 - 8.06 (m, 4 H), 7.54 - 7.69 (m, 2 H), 7.20 - 7.53 (m, 5 H), 5.14 - 5.56 (m, 4 H), 3.63 - 4.26 (m, 4 H), 3.1 1 and 3.30 (s, 3 H), 2.56 - 2.71 (m, 1 H), 1.93 - 2.40 (m, 1 H), 1.32 and 1.42 (s, 9 H);

LC-MS (ESI POS): 453.28 (M+).

EXAMPLE 24

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2- phenylacetoxy)-l-methyl-l-(2-oxo-2-(thiophen-2-yl)ethyl)pyrr olidinium bromide (C63)

Scheme 25

To a solution of (R)- l-methylpyrrolidin-3-yl 2-(tert- butoxycarbonylamino)-2-phenylacetate (C61) (170 mg, 0.51 mmol) in DMF (5 ml) and acetonitrile (3 ml), 2-bromo-l-(thiophen-2-yl)ethanone (115 mg, 0.56 mmol) was added and the reaction was stirred at RT for 15h. The solvent was evaporated and the crude was purified by flash chromatography (DCM/MeOH=9/l) to obtain (3R)-3-(2-(tert-butoxycarbonylamino)-2- phenylacetoxy)-l -methyl- l-(2-oxo-2-(thiophen-2-yl)ethyl)pyrrolidinium bromide (132 mg; 48% yield). 1H NMR (300 MHz, DMSO-<¾) δ ppm 8. 13 - 8.32 (m, 1 H), 7.98 - 8.08 (m, 1 H), 7.83 and 7.90 (d, 1 H), 7.13 - 7.53 (m, 6 H), 5.37 - 5.56 (m, 1 H), 5.03 - 5.37 (m, 3 H), 3.61 - 4. 19 (m, 4 H), 3. 10 and 3.24 (s, 3 H), 2.55 - 2.71 (m, 1 H), 1.91 - 2.38 (m, 1 H), 1.34 and 1.41 (s, 9 H);

LC-MS (ESI POS): 459.23 (M+).

EXAMPLE 25

Preparation of (3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2- (phenylsulfonamido)acetoxy)-l-azoniabicyclo [2.2.2] octane chloride (C65)

Scheme 26

Preparation of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate (130)

(R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-phenylacetate (C I) (5. 17 g, 14.3 mmol) was dissolved in THF (47.8 ml) and, while stirring at RT, 37% HC1 (4.71 ml, 57.4 mmol) was added dropwise. The reaction was stirred at RT for 15h overnight. The solvent was evaporated and the residue was dissolved in DCM/MeOH (9/1 ; 10 ml) and about 3g of SiO2 was added. The solvent was evaporated, the solid was loaded on a silica gel column and eluted with DCM/MeOH/NH ₄ OH (9/1/0.1) to obtain (R)-quinuclidin-3-yl 2-amino-2-phenylacetate (2.23 g; 60% yield).

Preparation of (R)-quinuclidin-3-yl 2-phenyl-2- (phenylsulfonamido)acetate (C64)

(R)-quinuclidin-3-yl 2-amino-2-phenylacetate (130) (100 mg, 0.38 mmol) was dissolved in DCM (4 ml) and TEA (0.11 ml, 0.77 mmol). Benzene sulfonyl chloride (59 μΐ, 0.46 mmol) was added and the solution was stirred at RT for 1 h. The volatiles were evaporated and the residue was purified by flash chromatography (DCM/MeOH/NH ₄ OH = 95/5/0.3) to obtain (R)-quinuclidin-3-yl 2-phenyl-2-(phenylsulfonamido)acetate (101 mg; 66% yield).

Preparation of (3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2- (phenylsulfonamido)acetoxy)-l-azoniabicyclo[2.2.2]octane chloride (C65) To a solution of (R)-quinuclidin-3-yl 2-phenyl-2-

(phenylsulfonamido)acetate (C64) (96 mg, 0.24 mmol) in EtOAc (1.6 ml) and acetonitrile (0.8 ml), 2-chloro- l-phenylethanone (40.8 mg, 0.26 mmol) was added. The solution was stirred at RT overnight. The solution was concentrated under vacuum and the crude was purified by flash chromatography (DCM/MeOH = 95/5 to 9/1) to obtain (3R)-l-(2-oxo-2- phenylethyl)-3-(2-phenyl-2-(phenylsulfonamido)acetoxy)- l- azoniabicyclo[2.2.2]octane chloride (99 mg; 74% yield).

1H NMR (300 MHz, DMSO-< ) δ ppm 9.06 (br. s., 1 H) 7.94 - 8.04 (m, 2 H) 7.71 - 7.83 (m, 3 H) 7.45 - 7.66 (m, 5 H) 7.24 - 7.41 (m, 5 H) 5.26 (d, 1 H) 5.22 (d, 1 H) 5.13 - 5.21 (m, 1 H) 4.95 - 5.11 (m, 1 H) 3.94 - 4.17 (m, 1 H) 3.47 - 3.79 (m, 5 H) 2.10 - 2.26 (m, 1 H) 1.69 - 2.08 (m, 4 H);

LC-MS (ESI POS): 519.27 (M+). EXAMPLE 26

Preparation of (3R)-l-(2-oxo-2-(thiazol-2-yl)ethyl)-3-(2-phenyl-2- (phenylsulfonamido)acetoxy)-l-azoniabicyclo[2.2.2]octane 2,2,2-

Scheme 27

Preparation of (R)-quinuclidin-3-yl 2-phenyl-2-

(phenylsulfonamido)acetate (C64):

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (0.67 g, 2.01 mmol) in DCM (25 ml), triethylamine (0.84 ml, 6.04 mmol) and benzene sulfonyl chloride (0.31 ml, 2.42 mmol) were sequentially added. The reaction was stirred at RT for 4h and then the solvent was evaporated. The residue was taken up with EtOAc and washed with water and brine, dried over Na ₂ SO ₄ and evaporated to obtain (R)-quinuclidin-3-yl 2- phenyl-2-(phenylsulfonamido)acetate (370 mg; 46% yield).

Preparation of (3R)-l-(2-oxo-2-(thiazol-2-yl)ethyl)-3-(2-phenyl-2- (phenylsulfonamido)acetoxy)-l-azoniabicyclo[2.2.2]octane 2,2,2- trifluoroacetate (C66)

To a solution of (R)-quinuclidin-3-yl 2-phenyl-2-

(phenylsulfonamido)acetate (C64) (90.0 mg, 0.22 mmol) in EtOAc (2 ml) and acetonitrile (5 ml), 2-bromo- l-(thiazol-2-yl)ethanone (50.9 mg, 0.25 mmol) was added. The reaction was stirred at RT for 39h then a second portion of 2-bromo-l-(thiazol-2-yl)ethanone (46.3 mg, 0.22 mmol) was added and the reaction is stirred at 60°C for 15h. The solvent was removed under reduced pressure and the resulting solid is first purified by flash chromatography (Hexane/EtOAc = 9/1) and then by preparative HPLC to obtain (3R)- l-(2-oxo- 2-(thiazol-2-yl)ethyl)-3-(2-phenyl-2-(phenylsulfonamido)acet oxy)-l- azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (13.6 mg; 10% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 8.99 (d, 1 H) 8.39 (t, 1 H) 8.24 (d, 1 H) 7.71 - 7.83 (m, 2 H) 7.44 - 7.64 (m, 3 H) 7.22 - 7.40 (m, 5 H) 5.10 - 5.22 (m, 3 H) 4.94 - 5.10 (m, 1 H) 3.97 - 4.14 (m, 1 H) 3.53 - 3.76 (m, 5 H) 2.12 - 2.23 (m, 1 H) 1.47 - 2.11 (m, 4 H);

LC-MS (ESI POS): 526.13 (M+).

EXAMPLE 27

Preparation of (3R)-l-(2-oxo-2-(thiophen-2-yl)ethyl)-3-(2-phenyl-2- (phenylsulfonamido)acetoxy)-l-azoniabicyclo[2.2.2]octane 2,2,2- trifluoroacetate (C67)

Scheme 28

To a solution of (R)-quinuclidin-3-yl 2-phenyl-2- (phenylsulfonamido)acetate (C64) (90.0 mg, 0.22 mmol) in EtOAc (2 ml) and acetonitrile (5 ml), 2-chloro-l-(thiophen-2-yl)ethanone (39.7 mg, 0.25 mmol) was added and the reaction was stirred at RT for 15h. Then 2-chloro-l- (thiophen-2-yl)ethanone (36.1 mg, 0.22 mmol) was added again and the reaction was heated at 60°C for 15h. The solvent was removed and crude was purified by flash chromatography (Hexane/EtOAc = 9/1) and then by preparative HPLC to obtain (3R)-l-(2-oxo-2-(thiophen-2-yl)ethyl)-3-(2- phenyl-2-(phenylsulfonamido)acetoxy)- l-azoniabicyclo[2.2.2]octane 2,2,2- trif uoroacetate (1 1.9 mg; 8% yield).

1H NMR (300 MHz, DMSO-d ₆ ) δ ppm 8.99 (d, 1 H), 8.20 and 8.21 (dd, 1 H), 8.05 and 8.07 (dd, 1 H), 7.70 - 7.85 (m, 2 H), 7.43 - 7.66 (m, 3 H), 7.22 - 7.39 (m, 6 H), 5. 12 - 5.21 (m, 1 H), 5.04 - 5.09 (m, 1 H), 4.99 and 5.02 (s, 2 H), 3.93 - 4. 13 (m, 1 H), 3.29 - 3.56 (m, 5 H), 2.1 1 - 2.23 (m, 1 H), 1.44 - 2. 10 (m, 4 H);

LC-MS (ESI POS): 525. 1 1 (M+).

EXAMPLE 28

Preparation of (3R)-3-(2-(4-methoxyphenylsulfonamido)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo [2.2.2]octane bromide (C69)

Scheme 29

Preparation of (R)-quinuclidin-3-yl 2-(4- methoxyphenylsulfonamido)-2-phenylacetate hydrochloride (C68):

(R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (150 mg, 0.45 mmol) was suspended in DCM (6 ml) and treated with triethylamine (99 μΐ, 1.35 mmol). Then 4-methoxybenzene- l-sulfonyl chloride (1 12 mg, 0.54 mmol) was added and reaction was stirred at RT for 3h. Solvent was evaporated and the crude was dissolved in EtOAc and washed with water and brine. The organic phase was dried over Na ₂ SO ₄ , filtered and evaporated. The crude was triturated with Et ₂ O to obtain (R)-quinuclidin-3-yl 2-(4-methoxyphenylsulfonamido)-2-phenylacetate hydrochloride (105 mg; 50% yield).

Preparation of (3R)-3-(2-(4-methoxyphenylsulfonamido)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane bromide (C69)

(R)-quinuclidin-3-yl 2-(4-methoxyphenylsulfonamido)-2-phenylacetate hydrochloride (C68) (96 mg, 0.22 mmol) was dissolved in DCM and washed with 1M K ₂ CO ₃ . The organic phase was dried over Na ₂ SO ₄ , filtered and evaporated. The resulting solid was dissolved in EtOAc (2 ml) and 2-bromo- l- phenylethanone (49.2 mg, 0.25 mmol) was added. The reaction was stirred at RT for 3h, then it was washed with aq. K ₂ CO ₃ , dried over Na ₂ SO ₄ filtered and evaporated to obtain (3R)-3-(2-(4-methoxyphenylsulfonamido)-2- phenylacetoxy)- l-(2-oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane bromide (75 mg; 53% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 8.84 (d, 1 H), 7.88 - 8.04 (m, 2

H), 7.66 - 7.82 (m, 3 H), 7.53 - 7.66 (m, 2 H), 7.25 - 7.44 (m, 5 H), 6.97 - 7. 1 1 (m, 2 H), 5. 12 - 5. 19 (m, 1 H), 4.95 - 5. 12 (m, 2 H), 3.96 - 4. 15 (m, 1 H), 3.81 (s, 3 H), 3.38 - 3.76 (m, 5 H), 2. 13 - 2.24 (m, 1 H), 1.74 - 2. 13 (m, 4 H);

LC-MS (ESI POS): 549. 19 (M+).

EXAMPLE 29

Preparation of (3R)-3-(2-(4-chlorophenylsulfonamido)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane bromide (C71)

Scheme 30

Preparation of (R)-quinuclidin-3-yl 2-(4-chlorophenylsulfonamido)- 2-phenylacetate (C70)

To a solution of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (150 mg, 0.45 mmol) in DCM (5 ml) and triethylamine (190 ul, 1.35 mmol), 4-chlorobenzene- l-sulfonyl chloride (114 mg, 0.54 mol) was added. The reaction was stirred at RT for 3h then the solvent was evaporated. The crude was taken up with EtOAc and washed with water, brine and then with K ₂ CO ₃ . The organic phase was dried over Na ₂ SO ₄ , filtered and evaporated to dryness to obtain (R)-quinuclidin-3-yl 2-(4-chlorophenylsulfonamido)-2-phenylacetate (62 mg; 32% yield), which was used as such in the next step.

Preparation of (3R)-3-(2-(4-chlorophenylsulfonamido)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane bromide (C71)

To a solution of (R)-quinuclidin-3-yl 2-(4-chlorophenylsulfonamido)-2- phenylacetate (C70) (62 mg, 0.14 mmol) in EtOAc (2.5 ml), 2-bromo-l- phenylethanone (31.2 mg, 0.16 mmol) was added and the reaction was stirred at RT for 3h. Then the solvent was evaporated and the crude was triturated with Et ₂ O to obtain (3R)-3-(2-(4-chlorophenylsulfonamido)-2-phenylacetoxy)- l-(2-oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane bromide (38 mg; 42% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 9.1 1 (d, 1 H), 7.88 - 8.07 (m, 2 H), 7.69 - 7.85 (m, 3 H), 7.51 - 7.69 (m, 4 H), 7.20 - 7.44 (m, 5 H), 5.17 - 5.26 (m, 1 H), 4.92 - 5.17 (m, 2 H), 3.95 - 4.19 (m, 1 H), 3.39 - 3.79 (m, 5 H), 2.15 - 2.25 (m, 1 H), 1.76 - 2.15 (m, 4 H);

LC-MS (ESI POS): 553.13 (M+).

EXAMPLE 30

Preparation of (3R)-3-(2-(3,4-difluorophenylsulfonamido)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane 2,2,2-trifluoroacetate (C73)

Scheme 31

Preparation of (R)-quinuclidin-3-yl 2-(3,4- difluorophenylsulfonamido)-2-phenylacetate (C72)

(R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (110 mg, 0.33 mmol) was suspended in DCM (4 ml) and triethylamine (121 μΐ, 1.65 mmol) was added obtaining a solution. 3,4-Difluorobenzene- l- sulfonyl chloride (53.1 μΐ, 0.40 mmol) was added and reaction was stirred at RT for 3h. The reaction was diluted with DCM and washed with water and brine. The organic phase was dried over Na ₂ SO ₄ , filtered and evaporated to dryness. The residue was triturated with Et ₂ O to obtain (R)-quinuclidin-3-yl 2-(3,4-difluorophenylsulfonamido)-2-phenylacetate (40 mg; 28% yield), which was used as such in the next step.

Preparation of (3R)-3-(2-(3,4-difluorophenylsulfonamido)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane 2,2,2-trifluoroacetate (C73)

To a solution of (R)-quinuclidin-3-yl

2-(3,4-difluorophenylsulfonamido)-2-phenylacetate (C72) (40 mg, 0.09 mmol) in EtOAc (3 ml), 2-bromo-l-phenylethanone (20.1 mg, 0.10 mmol) was added and reaction was stirred at RT for 2h. Then the solvent was evaporated and the crude was first triturated with petroleum ether and then purified by preparative HPLC to obtain (3R)-3-(2-(3,4-difluorophenylsulfonamido)-2-phenylacetoxy)- l-(2-oxo-2-phenylethyl)- l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (13 mg; 21% yield).

1H NMR (300 MHz, Acetone) δ ppm 8.89 and 8.96 (d, 1 H), 8.00 - 8.16

(m, 2 H), 7.18 - 7.91 (m, 11 H), 5.52 and 5.58 (d, 1 H), 5.44 and 5.51 (d, 1 H), 5.28 and 5.35 (d, 1 H), 5.18 - 5.29 (m, 1 H), 4.29 - 4.49 (m, 2 H), 3.75 - 4.28 (m, 4 H), 2.39 - 2.53 (m, 1 H), 2.11 - 2.36 (m, 3 H), 1.69 - 2.03 (m, 1 H);

LC-MS (ESI POS): 555.12 (M+).

EXAMPLE 31

Preparation of (3R)-3-(2-(2,4-dimethylthiazole-5-sulfonamido phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane 2,2,2-trifluoroacetate (C75)

Scheme 32

Preparation of (R)-quinuclidin-3-yl 2-(2,4-dimethylthiazole-5- sulfonamido)-2-phenylacetate (C74)

(R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydro chloride (125)

(116 mg, 0.35 mmol) was suspended in DCM (6 ml) and triethylamine (102 μΐ, 1.39 mmol) was added obtaining a clear solution. 2,4-Dimethylthiazole-5-sulfonyl chloride (88.0 mg, 0.42 mmol) was added and reaction was stirred ad RT for 3h. Reaction was diluted with DCM and washed with water and brine. The organic phase was dried with Na ₂ SO ₄ , filtered and evaporated to dryness. The crude was purified by flash chromatography (DCM/MeOH = 85/15) to obtain (R)-quinuclidin-3-yl 2-(2,4- dimethylthiazole-5-sulfonamido)-2-phenylacetate (67 mg; 44.2% yield).

Preparation of (3R)-3-(2-(2,4-dimethylthiazole-5-sulfonamido)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane 2,2,2-trifluoroacetate (C75):

To a solution of (R)-quinuclidin-3-yl 2-(2,4-dimethylthiazole-5- sulfonamido)-2-phenylacetate (C74) (67.0 mg, 0.15 mmol) in EtOAc (2 ml), 2-bromo-l-phenylethanone (33.7 mg, 0.17 mmol) was added. The reaction was stirred at RT for 2h. The solvent was evaporated and the crude was purified by preparative HPLC obtaining (3R)-3-(2-(2,4-dimethylthiazole-5- sulfonamido)-2-phenylacetoxy)- l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (18 mg; 18% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 9.43 and 9.44 (d, 1 H), 7.89 - 8.05 (m, 2 H), 7.70 - 7.82 (m, 1 H), 7.56 - 7.69 (m, 2 H), 7.24 - 7.46 (m, 5 H), 5.15 - 5.26 (m, 2 H), 5.13 (s, 2 H), 3.94 - 4.21 (m, 1 H), 3.43 - 3.62 (m, 5 H), 2.55 and 2.58 (s, 3 H), 2.39 and 2.42 (s, 3 H), 2.06 - 2.20 and 2.20 - 2.30 (m, 1 H), 1.47 - 2.05 (m, 4 H);

LC-MS (ESI POS): 553.13 (M+).

EXAMPLE 32

Preparation of (3R)-3-(2-(methylsulfonamido)-2-phenylacetoxy)-l- (2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide (C77)

Scheme 33

Preparation of (R)-quinuclidin-3-yl 2-(methylsulfonamido)-2- phenylacetate (C76):

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (120 mg, 0.36 mmol) in DCM (6 ml), triethylamine (79 μΐ, 1.08 mmol) and methane sulfonyl chloride (33.4 μΐ, 0.43 mmol) were sequentially added. The reaction was stirred at RT for 3h then the volatiles were evaporated. The crude was taken up with EtOAc and washed with water and brine. The organic layer was dried over Na ₂ SO ₄ , filtered and evaporated to dryness. The crude was triturated with Et ₂ O to obtain (R)-quinuclidin-3-yl 2- (methylsulfonamido)-2-phenylacetate (38.9 mg; 32% yield).

Preparation of (3R)-3-(2-(methylsulfonamido)-2-phenylacetoxy)-l- (2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide (C77)

To a solution of (R)-quinuclidin-3-yl 2-(methylsulfonamido)-2- phenylacetate (C76) (38.9 mg, 0.11 mmol) in EtOAc (1 ml) and acetonitrile (0.5 ml), 2-bromo- l-phenylethanone (25.2 mg, 0.13 mmol) was added. The mixture was stirred at RT for 2h then the solvent was evaporated. The crude was triturated with Et ₂ O to obtain (3R)-3-(2-(methylsulfonamido)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane bromide (40 mg; 64.7% yield).

1H NMR (300 MHz, DMSO- ₆ ) δ ppm 8.37 (d, 1 H), 7.90 - 8.05 (m, 2 H), 7.69 - 7.83 (m, 1 H), 7.55 - 7.67 (m, 2 H), 7.29 - 7.55 (m, 5 H), 5.27 - 5.34 (m, 1 H), 5.20 - 5.27 (m, 1 H), 5.10 - 5.20 (m, 2 H), 4.00 - 4.23 (m, 1 H), 3.46 - 3.81 (m, 5 H), 2.89 and 2.93 (s, 3 H), 2,22 and 2.39 (m, 1 H), 1.50 - 2.15 (m, 4 H);

LC-MS (ESI POS): 457.17 (M+).

EXAMPLE 33

Preparation of (3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2-(2,2,2- trifluoroethylsulfonamido)acetoxy)-l-azoniabicyclo [2.2.2] octane bromide (C79)

Scheme 34

Preparation of (R)-quinuclidin-3-yl 2-phenyl-2-(2,2,2- trifluoroethylsulfonamido)acetate (C78)

solution of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (120 mg, 0.36 mmol) in DCM (4 ml) and triethylamine (105 μΐ, 1.44 mmol), 2,2,2-trifluoroethanesulfonyl chloride (47.5 μΐ, 0.43 mmol) was added. The reaction was stirred at RT for 3h and then it was diluted with DCM and washed with water, brine and 1M K ₂ CO ₃ . The organic phase was dried with Na ₂ SO ₄ , filtered and evaporated. The crude was purified with flash chromatography (DCM/MeOH = 9/1) to obtain (R)-quinuclidin-3-yl 2-phenyl-2-(2,2,2-trifluoroethylsulfonamido)acetate (60 mg; 41% yield).

Preparation of (3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2-(2,2,2- trifluoroethylsulfonamido)acetoxy)-l-azoniabicyclo[2.2.2]oct ane bromide (C79)

To a solution of (R)-quinuclidin-3-yl 2-phenyl-2-(2,2,2- trifluoroethylsulfonamido)acetate (C78) (60 mg, 0.15 mmol) in EtOAc (2 ml), 2-bromo-l-phenylethanone (35.3 mg, 0.18 mmol) was added. The reaction was stirred at RT for 16h. Then the solvent was evaporated and the crude was purified by flash chromatography (DCM/MeOH=9/l) to obtain (3R)- l-(2-oxo- 2-phenylethyl)-3-(2-phenyl-2-(2,2,2-trifluoroethylsulfonamid o)acetoxy)- l- azoniabicyclo[2.2.2]octane bromide (5 mg; 6% yield).

1H NMR (300 MHz, Acetonitrile- ₃ ) δ ppm 7.91 - 8.06 (m, 2 H), 7.71 - 7.83 (m, 1 H), 7.67 (d, 1 H), 7.56 - 7.64 (m, 2 H), 7.37 - 7.56 (m, 5 H), 5.36 and 5.37 (d, 1 H), 5.16 - 5.31 (m, 1 H), 4.91 and 4.98 (d, 1 H), 4.84 and 4.91 (d, 1 H), 3.94 - 4.33 (m, 3 H), 3.50 - 3.93 (m, 5 H), 2.32 - 2.42 and 2.42 - 2.55 (m, 1 H), 2.00 - 2.23 (m, 3 H), 1.79 - 1.92 (m, 1 H);

LC-MS (ESI POS): 525.10 (M+). EXAMPLE 34

Preparation of ((3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2- (phenylmethylsulfonamido)acetoxy)-l-azoniabicyclo[2.2.2]octa ne bromide (C81)

Scheme 35

Preparation of (R)-quinuclidin-3-yl 2-phenyl-2-

(phenylmethylsulfonamido)acetate (C80)

To a solution of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (100 mg, 0.30 mmol) in DCM (4 ml) and triethylamine (65.8 μΐ, 0.90 mmol), phenylmethanesulfonyl chloride (68.6 mg, 0.36 mmol) was added. The reaction was stirred at RT for 13h. DCM was evaporated and the crude was taken up with EtOAc and washed 1M Na ₂ CO ₃ , water and brine. The organic phase was dried over Na ₂ SO ₄ , filtered and evaporated to obtain (R)-quinuclidin-3-yl 2-phenyl-2-(phenylmethylsulfonamido)acetate (74 mg; 59.5% yield).

Preparation of (3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2- (phenylmethylsulfonamido)acetoxy)-l-azoniabicyclo[2.2.2]octa ne bromide (C81)

To a solution of (R)-quinuclidin-3-yl 2-phenyl-2- (phenylmethylsulfonamido)acetate (C80) (41 mg, 0.10 mmol) in EtOAc (2 ml), 2-bromo- l-phenylethanone (21.7 mg, 0.11 mmol) was added and the reaction was stirred at RT for 1 h. The solvent was evaporated and the crude was triturated with Et ₂ O to obtain (3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl- 2-(phenylmethylsulfonamido)acetoxy)- l-azoniabicyclo[2.2.2]octane bromide (50 mg; 82% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 8.42 (d, 1 H), 7.88 - 8.07 (m, 2 H), 7.69 - 7.81 (m, 1 H), 7.53 - 7.67 (m, 2 H), 7.24 - 7.52 (m, 10 H), 5.18 - 5.33 (m, 1 H), 5.08 - 5.14 (m, 1 H), 5.13 (d, 1 H), 4.44 (d, 1 H), 4.37 (d, 1 H), 4.02 - 4.21 (m, 1 H), 3.45 - 3.84 (m, 5 H), 2.32 - 2.42 (m, 1 H), 1.43 - 2.15 (m, 4 H);

LC-MS (ESI POS): 533.23 (M+).

EXAMPLE 35

Preparation of (3R)-3-(2-(4-fluorophenyl)-2-

(phenylsulfonamido)acetoxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (C84)

C84

Scheme 36 Preparation of (R)-quinuclidin-3-yl 2-amino-2-(4- fluorophenyl)acetate dihydrochloride (182)

To a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2- (4-fluorophenyl)acetate (C50) (350 mg, 0.92 mmol) in THF (20 ml), 37% hydrogen chloride (1.0 ml, 12.2 mmol) was added and the reaction was stirred at RT for 15h. Then a second portion of 37% hydrogen chloride (1.0 ml, 12.2 mmol) was added again and the reaction was further stirred at RT for 24h. Then the volatiles were evaporated under vacuum to obtain (R)-quinuclidin-3 -yl 2-amino-2-(4-fluorophenyl)acetate dihydrochloride (325 mg; quantitative yield).

Preparation of (R)-quinuclidin-3-yl 2-(4-fluorophenyl)-2- (phenylsulfonamido)acetate (C83)

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-(4- fluorophenyl)acetate dihydrochloride (182) (325 mg, 0.92 mmol) in DCM (10 ml), triethylamine (386 μΐ, 2.78 mmol) and benzene sulfonyl chloride (142 μΐ, 1.11 mmol) were added. The reaction was stirred at RT for 4h and then the solvent was evaporated. The residue was taken up with EtOAc and washed with 1M Na ₂ CO ₃ , water and then brine, dried over Na ₂ SO ₄ and evaporated to dryness to obtain (R)-quinuclidin-3-yl 2-(4-fluorophenyl)-2- (phenylsulfonamido)acetate (150 mg; 39% yield).

Preparation of (3R)-3-(2-(4-fluorophenyl)-2-

(phenylsulfonamido)acetoxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (C84)

To a solution of (R)-quinuclidin-3-yl 2-(4-fluorophenyl)-2- (phenylsulfonamido)acetate (C83) (150 mg, 0.36 mmol) in EtOAc (2 ml) and acetonitrile (2 ml), 2-bromo-l-phenylethanone (78 mg, 0.39 mmol) was added. The reaction was stirred at RT for 15h. The solvent was evaporated and the crude was purified by preparative HPLC to obtain (3R)-3-(2-(4-fluorophenyl)- 2-(phenylsulfonamido)acetoxy)- l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (97.9 mg; 42% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 9.02 (d, 1 H), 7.89 - 8.09 (m, 2 H), 7.66 - 7.81 (m, 3 H), 7.32 - 7.68 (m, 7 H), 7.02 - 7.21 (m, 2 H), 5.20 - 5.30 (m, 1 H), 5.11 and 5.14 (s, 2 H), 4.94 - 5.10 (m, 1 H), 3.94 - 4.14 (m, 1 H), 3.38 - 3.66 (m, 5 H), 2.14 - 2.25 (m, 1 H), 1.53 - 2.09 (m, 4 H);

LC-MS (ESI POS): 537.18 (M+).

EXAMPLE 36

Preparation of (3R)-3-(2-benzamido-2-phenylacetoxy)-l-(2-oxo-2- phenylethyl)-l-azoniabicyclo[2.2.2]octane chloride (C86)

Scheme 37

Preparation of (R)-quinuclidin-3-yl 2-benzamido-2-phenylacetate (C85) To a solution of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate (130)

(100 mg, 0.38 mmol) in DCM (4 ml) and triethylamine (0.08 ml, 0.58 mmol), benzoyl chloride (58.0 μΐ, 0.50 mmol) was added. The reaction was stirred at RT for 1.5h and then the volatiles were evaporated. The crude was purified by flash chromatography (DCM/MeOH/NH ₄ OH =95/5/0.3) to obtain (R)-quinuclidin-3-yl 2-benzamido-2-phenylacetate (70 mg; 50% yield).

Preparation of (3R)-3-(2-benzamido-2-phenylacetoxy)-l-(2-oxo-2- phenylethyl)-l-azoniabicyclo[2.2.2]octane chloride (C86)

To a solution of (R)-quinuclidin-3-yl 2-benzamido-2-phenylacetate (C85) (70 mg, 0.19 mmol) in EtOAc (1 ml) and acetonitrile (1 ml), 2-chloro- 1- phenylethanone (32.7 mg, 0.21 mmol) was added. The reaction was stirred at RT for 24h, then the solvents were evaporated and the residue was triturated with EtOAc (8 ml) to obtain (3R)-3-(2-benzamido-2-phenylacetoxy)-l-(2-oxo-

2- phenylethyl)-l-azoniabicyclo[2.2.2]octane chloride (72 mg; 72% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 9.38 (d, J=7.04 Hz, 1 H), 7.88 - 8.11 (m, 4 H), 7.69 - 7.87 (m, 1 H), 7.12 - 7.68 (m, 10 H), 5.78 (d, J=7.04 Hz, 1 H), 5.26 (s, 2 H), 5.15 - 5.24 (m, 1 H), 4.05 - 4.27 (m, 1 H), 3.81 - 3.94 (m, 1 H), 3.54 - 3.81 (m, 4 H), 2.19 - 2.3 1 (m, 1 H), 1.91 - 2.14 (m, 2 H), 1.74 - 1.91 (m, 1 H), 1.61 - 1.74 (m, 1 H);

LC-MS (ESI POS): 483.02 (M+).

EXAMPLE 37

Preparation of (3R)-3-(2-benzamido-2-phenylacetoxy)-l-(2-(4- fluorophenyl)-2-oxoethyl)-l-azoniabicyclo [2.2.2] octane 2,2,2- trifluoroacetate (C87)

Scheme 38

Preparation of (R)-quinuclidin-3-yl 2-benzamido-2-phenylacetate (C85)

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (0.67 g, 2.01 mmol) in DCM (25 ml), triethylamine (0.84 ml, 6.04 mmol) and benzoyl chloride (0.28 ml, 2.42 mmol) were sequentially added. The reaction was stirred at RT for 4h then the solvent was evaporated. The crude was taken up with EtOAc and washed with water and brine, dried over Na ₂ SO ₄ and evaporated to obtain (R)-quinuclidin-3-yl 2-benzamido-2-phenylacetate (310 mg; 42% yield).

Preparation of (3R)-3-(2-benzamido-2-phenylacetoxy)-l-(2-(4- fluorophenyl)-2-oxoethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2- trifluoroacetate (C87)

To a solution of (R)-quinuclidin-3-yl 2-benzamido-2-phenylacetate (C85)

(77.5 mg, 0.21 mmol) in EtOAc (1 ml) and acetonitrile (5 ml), 2-chloro-l-(4- fluorophenyl)ethanone (40.4 mg, 0.23 mmol) was added. The reaction was stirred at RT for 15h and then the solvent was evaporated. The crude was triturated with z-Pr ₂ O/EtOAc (5/1) and then purified by preparative HPLC to obtain (3R)-3-(2- benzamido-2-phenylacetoxy)-l-(2-(4-fluorophenyl)-2-oxoethyl) -l- azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (50.8 mg; 39% yield).

1H NMR (300 MHz, DMSO-< ) δ ppm 9.27 (d, 1 H), 7.99 - 8.16 (m, 2 H), 7.84 - 7.99 (m, 2 H), 7.28 - 7.67 (m, 10 H), 5.66 - 5.84 (m, 1 H), 5.26 (m, 1 H), 5.14 and 5.15 (s, 2 H), 3.97 - 4.30 (m, 1 H), 3.36 - 3.73 (m, 5 H), 2.18 - 2.26 and 2.38 - 2.46 (m, 1 H), 1.57 - 2.17 (m, 4 H);

LC-MS (ESI POS): 501.08 (M+).

The compounds listed in Table 5 were obtained as previously described for C87, starting from compound C85 and the suitable commercially available alkylating agents.

Table 5

(continued)

(continued)

EXAMPLE 38

Preparation of (3R)-3-(2-acetamido-2-phenylacetoxy)-l-(2-oxo-2 ylethyl)-l-azoniabicyclo [2.2.2] octane 2,2,2-trifluoroacetate (C93)

Scheme 39

Preparation of (R)-quinuclidin-3-yl 2-acetamido-2-phenylacetate

(C92)

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (140 mg, 0.42 mmol) in DCM (10 ml), triethylamine (175 μΐ, 1.26 mmol) and acetyl chloride (35.8 μΐ, 0.50 mmol) were sequentially added. The reaction was stirred at RT for 2h then the solvent was evaporated. The residue was taken up with little EtOAc and the insoluble was filtered off. The organic phase was evaporated to dryness achieving (R)- quinuclidin-3-yl 2-acetamido-2-phenylacetate ( 127 mg; quantitative yield).

Preparation of (3R)-3-(2-acetamido-2-phenylacetoxy)-l-(2-oxo-2- phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (C93)

To a solution of (R)-quinuclidin-3-yl 2-acetamido-2-phenylacetate (C92) (127 mg, 0.42 mmol) in EtOAc (2 ml) and acetonitrile (5 ml), 2-bromo-l- phenylethanone (92 mg, 0.46 mmol) was added and the reaction was stirred at RT for 72h. The solvent was evaporated and the crude was purified by preparative HPLC to obtain (3R)-3-(2-acetamido-2-phenylacetoxy)-l-(2-oxo-2-phenylethyl) - l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (20.5 mg; 9% yield).

1H NMR (300 MHz, DMSO-< ) δ ppm 8.77 and 8.79 (d, 1 H), 7.89 - 8.04 (m, 2 H), 7.70 - 7.82 (m, 1 H), 7.55 - 7.69 (m, 2 H), 7.29 - 7.54 (m, 5 H), 5.44 and 5.48 (d, 1 H), 5.18 - 5.27 (m, 1 H), 5.15 and 5. 18 (s, 2 H), 3.98 - 4.21 (m, 1 H), 3.52 - 3.81 (m, 5 H), 1.94 (s, 3 H), 1.58 - 2.45 (m, 5 H);

LC-MS (ESI POS): 421. 16 (M+).

EXAMPLE 39

Preparation of (3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2- pivalamidoacetoxy)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (C95)

Scheme 40 Preparation of (R)-quinuclidin-3-yl 2-phenyl-2-pivalamidoacetate

(C94)

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (125) (140 mg, 0.42 mmol) in DCM (5 ml), triethylamine (175 μΐ, 1.26 mmol) and pivaloyl chloride (62.1 μΐ, 0.50 mmol) were sequentially added. The reaction was stirred at RT for 2h and then the solvent was evaporated. The residue was taken up with EtOAc and washed with Na ₂ CO ₃ , water and brine, dried over Na ₂ SO ₄ , filtered and evaporated to obtain (R)-quinuclidin-3-yl 2-phenyl-2-pivalamidoacetate (95 mg; 66% yield).

Preparation of (3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2- pivalamidoacetoxy)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (C95)

To a solution of (R)-quinuclidin-3-yl 2-phenyl-2-pivalamidoacetate (C94) (95 mg, 0.28 mmol) in EtOAc (2 ml) and acetonitrile (2 ml), 2-bromo- 1-phenylethanone (60.4 mg, 0.30 mmol) was added and the reaction was stirred at RT for 15h. The solvent was evaporated and the crude was purified by preparative HPLC to obtain (3R)-l-(2-oxo-2-phenylethyl)-3-(2-phenyl-2- pivalamidoacetoxy)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (118.6 mg; 75% yield).

1H NMR (300 MHz, DMSO-< ) δ ppm 8.21 (t, 1 H) 7.91 - 8.05 (m, 2 H)

7.69 - 7.83 (m, 1 H) 7.55 - 7.69 (m, 2 H) 7.26 - 7.54 (m, 5 H) 5.40 - 5.54 (m, 1 H) 5.16 (d, 2 H) 5.08 - 5.34 (m, 1 H) 4.01 - 4.34 (m, 3 H) 3.43 - 3.79 (m, 3 H) 2.32 - 2.45 (m, 1 H) 1.87 - 2.16 (m, 3 H) 1.74 (s, 1 H) 1.17 (s, 9 H);

LC-MS (ESI POS): 463.19 (M+).

The compounds listed in Table 6 were obtained as previously described for C95, treating intermediate 125 with the suitable commercially available acyl halides, followed by quaternization with 2 -bromo- 1-phenylethanone and purification by preparative HPLC. Table 6

(continued)

(continued)

(continued) EXAMPLE 40

Preparation of (3R)-3-(2-benzamido-2-(4-methoxyphenyl)acetoxy)- l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2- trifluoroacetate (C108)

Scheme 41

Preparation of (R)-quinuclidin-3-yl 2-amino-2-(4- methoxyphenyl)acetate dihydrochloride (1106)

To a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-

(4-methoxyphenyl)acetate (C44) (560 mg, 1.43 mmol) in THF (20 ml), 37% hydrogen chloride (1.0 ml, 12.2 mmol) was added. The reaction was stirred at RT for 15h. Then a second portion of 37% hydrogen chloride (1.0 ml, 12.2 mmol) was added and the reaction was stirred at RT for additional 48h. The solvent was evaporated to obtain (R)-quinuclidin-3-yl 2-amino-2-(4- methoxyphenyl)acetate dihydrochloride (521 mg; quantitative yield).

Preparation of (R)-quinuclidin-3-yl 2-benzamido-2-(4- methoxyphenyl)acetate (C107)

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-(4- methoxyphenyl)acetate dihydro chloride (1106) (325 mg, 0.89 mmol) in DCM (10 ml), triethylamine (374 μΐ, 2.68 mmol) and benzoyl chloride (125 μΐ, 1.07 mmol) were added. The reaction was stirred at RT for 4h then the solvent was evaporated. The residue was taken up with EtOAc and washed with Na ₂ CO ₃ , water and brine. The organic phase was dried over Na ₂ SO ₄ , filtered and evaporated to obtain (R)-quinuclidin-3-yl 2-benzamido-2-(4- methoxyphenyl)acetate (145 mg; 41% yield). The product was used in the next step without any further purification.

Preparation of (3R)-3-(2-benzamido-2-(4-methoxyphenyl)acetoxy)- l-(2-oxo-2-phenylethyl)-l-azoniabicyclo [2.2.2] octane 2,2,2- trifluoroacetate (C108)

To a solution of (R)-quinuclidin-3-yl 2-benzamido-2-(4- methoxyphenyl)acetate (C 107) (145 mg, 0.37 mmol) in EtOAc (2 ml) and acetonitrile (2 ml), 2-bromo-l-phenylethanone (80 mg, 0.40 mmol) was added and the reaction was stirred at RT for 15h. The solvent was evaporated and the crude was purified by preparative HPLC to obtain (3R)-3-(2-benzamido-2-(4- methoxyphenyl)acetoxy)- l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (52.4 mg; 23% yield).

1H NMR (300 MHz, DMSO-< ) δ ppm 9.16 (d, 1 H), 7.87 - 8.07 (m, 4 H), 7.69 - 7.82 (m, 1 H), 7.53 - 7.69 (m, 3 H), 7.38 - 7.53 (m, 4 H), 6.89 - 7.10 (m, 2 H), 5.50 - 5.69 (m, 1 H), 5.19 - 5.29 (m, 1 H), 5.16 (s, 2 H), 4.02 - 4.26 (m, 1 H), 3.78 (s, 3 H), 3.50 - 3.75 (m, 5 H), 2.36 - 2.47 (m, 1 H), 1.66 - 2.23 (m, 4 H);

LC-MS (ESI POS): 513.12 (M+).

EXAMPLE 41

Preparation of (3R)-3-(2-benzamido-2-p-tolylacetoxy)-l-(2-oxo-2- phenylethyl)-l-azoniabicyclo [2.2.2] octane 2,2,2-trifluoroacetate (CI 11)

EtOAc-CH ₃ CN

Scheme 42

Preparation of (R)-quinuclidin-3-yl 2-amino-2-p-tolylacetate dihydrochloride (1109)

To a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2- p-tolylacetate (C41) (250 mg, 0.67 mmol) in THF (50 ml), 37% hydrogen chloride (1.0 ml, 12.2 mmol) was added and the reaction was stirred at RT for 15h. The solvent was evaporated to obtain (R)-quinuclidin-3-yl 2-amino-2-p- tolylacetate dihydrochloride (232 mg; quantitative yield).

Preparation of (R)-quinuclidin-3-yl 2-benzamido-2-p-tolylacetate (C110)

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-p-tolylacetate dihydrochloride (1109) (1 16 mg, 0.33 mmol) in DCM (5 ml), triethylamine (139 μΐ, 1.00 mmol) and benzoyl chloride (46.5 μΐ, 0.40 mmol) were added. The reaction was stirred at RT for 3h then the solvent was evaporated. The residue was taken up with EtOAc and washed with Na ₂ CO ₃ , water and brine. The organic phase was dried over Na ₂ SO ₄ , filtered and evaporated to obtain (R)-quinuclidin-3-yl 2-benzamido-2-p-tolylacetate (70.0 mg; 55% yield). The product was used in the next step without any further purification.

Preparation of (3R)-3-(2-benzamido-2-p-tolylacetoxy)-l-(2-oxo-2- phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (Oi l)

2-Bromo-l-phenylethanone (40.5 mg, 0.20 mmol) was added to a solution of (R)-quinuclidin-3-yl 2-benzamido-2-p-tolylacetate (C I 10) (70.0 mg, 0.18 mmol) in EtOAc (3 ml). The reaction was stirred at RT for 15h, then 2-bromo- l-phenylethanone (36.8 mg, 0.18 mmol) was added again and the reaction was stirred at RT for 6h. The solvent was evaporated and the crude was purified by preparative HPLC to obtain (3R)-3-(2-benzamido-2-p- tolylacetoxy)- l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2- trifluoroacetate (54.8 mg; 48% yield).

1H NMR (300 MHz, DMSO-<¾) δ ppm 9.19 and 9.20 (d, 1 H), 7.86 - 8.05 (m, 4 H), 7.71 - 7.84 (m, 1 H), 7.52 - 7.71 (m, 3 H), 7.39 - 7.52 (m, 4 H), 7.17 - 7.33 (m, 2 H), 5.65 and 5.69 (d, 1 H), 5.21 - 5.31 (m, 1 H), 5.16 and 5.18 (s, 2 H), 4.00 - 4.26 (m, 1 H), 3.38 - 3.89 (m, 5 H), 2.19 - 2.27 and 2.38 - 2.46 (m, 1 H), 2.33 (s, 3 H), 1.48 - 2.17 (m, 4 H);

LC-MS (ESI POS): 497.12 (M+).

EXAMPLE 42

Preparation of (3R)-3-(2-benzamido-2-(4-chlorophenyl)acetoxy)-l- (2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (C114)

Scheme 43

Preparation of (R)-quinuclidin-3-yl 2-amino-2-(4- chlorophenyl)acetate dihydrochloride (1112)

To a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-

(4-chlorophenyl)acetate (C47) (250 mg, 0.63 mmol) in THF (5 ml), 37% hydrogen chloride (1.0 ml, 12.2 mmol) was added. The reaction was stirred at RT for 15h. The solvent was evaporated to obtain (R)-quinuclidin-3-yl 2-amino-2-(4-chlorophenyl)acetate dihydrochloride (233 mg; quantitative yield). Preparation of (R)-quinuclidin-3-yl 2-benzamido-2-(4- chlorophenyl)acetate (C113)

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-(4- chlorophenyl)acetate dihydrochloride (1112) (0.12 g, 0.32 mmol) in DCM (5 ml), triethylamine (0.13 ml, 0.95 mmol) and benzoyl chloride (44.0 μΐ, 0.38 mmol) were added and the reaction was stirred at RT for 3h. Then the solvent was evaporated and residue was taken up with EtOAc and washed with Na ₂ CO ₃ , water and brine. The organic layer was dried over Na ₂ SO ₄ , filtered and evaporated to obtain (R)-quinuclidin-3-yl 2-benzamido-2-(4- chlorophenyl)acetate (0.126 g; quantitative yield).

Preparation of (3R)-3-(2-benzamido-2-(4-chlorophenyl)acetoxy)-l- (2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (C114)

To a solution of (R)-quinuclidin-3-yl 2-benzamido-2-(4- chlorophenyl)acetate (C I 13) (126 mg, 0.32 mmol) in EtOAc (3 ml), 2-bromo- 1-phenylethanone (69.2 mg, 0.35 mmol) was added. The reaction was stirred at RT for 15h, then a second portion of 2-bromo-l-phenylethanone (62.9 mg, 0.32 mmol) was added and the reaction was stirred for additional 6h. The solvent was evaporated and the crude was purified by preparative HPLC to obtain (3R)-3-(2-benzamido-2-(4-chlorophenyl)acetoxy)-l-(2-oxo-2- phenylethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (46.0 mg; 23% yield).

1H NMR (300 MHz, DMSO-< ) δ ppm 9.29 and 9.31 (d, 1 H), 7.87 - 8.07 (m, 4 H), 7.71 - 7.82 (m, 1 H), 7.54 - 7.68 (m, 5 H), 7.44 - 7.54 (m, 4 H), 5.76 and 5.78 (d, 1 H), 5.21 - 5.31 (m, 1 H), 5.16 and 5.17 (br. s., 2 H), 3.97 - 4.25 (m, 1 H), 3.43 - 3.93 (m, 5 H), 2.20 - 2.31 and 2.33 - 2.46 (m, 1 H), 1.58 - 2.18 (m, 4 H);

LC-MS (ESI POS): 517.12 (M+). EXAMPLE 43

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2-(thiophen- 2-yl)acetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2] octane chloride (C116)

C115 C116

Scheme 44

Preparation of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)- 2-(thiophen-2-yl)acetate (CI 15)

A mixture of 2-(tert-butoxycarbonylamino)-2-(thiophen-2-yl)acetic acid

(710 mg, 2.76 mmol), (R)-quinuclidin-3-ol (421 mg, 3.31 mmol), DCC (683 mg, 3.31 mmol), HOBT (507 mg, 3.31 mmol) in THF (20 ml) was stirred at RT overnight. THF was evaporated, the crude was portioned between EtOAc and 1M K ₂ CO ₃ . The organic phase was dried over Na ₂ SO ₄ , filtered and evaporated to dryness. The crude was purified by flash chromatography (DCM/MeOH=98/2) to obtain (R)-quinuclidin-3-yl 2-(tert- butoxycarbonylamino)-2-(thiophen-2-yl)acetate (509 mg, 50.3% yield).

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-2-(thiophen- 2-yl)acetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2] octane chloride (C116)

2-Chloro-l-phenylethanone (43.0 mg, 0.28 mmol) was added to a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-2-(thiophen-2- yl)acetate (102 mg, 0.28 mmol) in EtOAc (3 ml). The reaction was stirred at RT for 20 hours. The solvent was evaporated and the crude was triturated with Et ₂ O (2 ml) and dried under vacuum to obtain (3R)-3-(2-(tert- butoxycarbonylamino)-2-(thiophen-2-yl)acetoxy)- l-(2-oxo-2-phenylethyl)- l- azoniabicyclo[2.2.2]octane chloride (70 mg, 48.3% yield).

1H NMR (300 MHz, DMSO-d6) δ ppm 8.02 - 8.07 (m, 1 H), 7.87 - 8.02 (m, 2 H), 7.68 - 7.84 (m, 1 H), 7.56 - 7.68 (m, 2 H), 7.52 (dd, 1 H), 7.12 - 7.26 (m, 1 H), 7.03 (dd, 1 H), 5.42 - 5.65 (m, 1 H), 5. 17 - 5.33 (m, 1 H), 5.21 (s, 2 H), 4.07 - 4.29 (m, 1 H), 3.47 - 3.85 (m, 5 H), 2.21 - 2.33 and 2.35 - 2.45 (m, 1 H), 1.97 - 2. 19 (m, 2 H), 1.66 - 1.97 (m, 2 H), 1.42 (s, 9 H);

LC-MS (ESI POS): 485.09 (M+).

EXAMPLE 44

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-3-(lH-indol- 3-yl)propanoyloxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2 .2.2]octane bromide (C118)

C117 C118

Scheme 45

Preparation of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)- 3-(lH-indol-3-yl)propanoate (C117)

A solution of 2-(tert-butoxycarbonylamino)-3-(lH-indol-3-yl)propanoic acid (100 mg, 0.33 mmol), (R)-quinuclidin-3-ol (50.1 mg, 0.39 mmol), HOBT (53.3 mg, 0.39 mmol) and EDC (76 mg, 0.39 mmol) in DMF (2 ml) was heated at 90°C for 1.5h under microwave irradiation. The solvent was evaporated. The residue was taken up with EtOAc and washed with IN Na ₂ CO ₃ , water and brine. The organic phase was dried over Na ₂ SO ₄ , filtered and evaporated to obtain (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)- 3-(lH-indol-3-yl)propanoate (95 mg, 69.9% yield), which was used as such in the next step.

Preparation of (3R)-3-(2-(tert-butoxycarbonylamino)-3-(lH-indol- 3-yl)propanoyloxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo [2.2.2] octane bromide (C118)

To a solution of (R)-quinuclidin-3-yl 2-(tert-butoxycarbonylamino)-3- (lH-indol-3-yl)propanoate (95 mg, 0.23 mmol) in EtOAc (3 ml), 2-bromo-l- phenylethanone (54.9 mg, 0.28 mmol) was added and the reaction was stirred at RT for 15h. The solid was collected by filtration and purified by silica gel flash chromatography (DCM/MeOH =94/6) to obtain (3R)-3-(2-(tert- butoxycarbonylamino)-3-(lH-indol-3-yl)propanoyloxy)- l-(2-oxo-2- phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide (31.0 mg, 22% yield).

1H NMR (300 MHz, DMSO-d6) δ ppm 10.87 (br. s., 1 H), 7.93 - 8.05 (m, 2 H), 7.70 - 7.81 (m, 1 H), 7.58 - 7.69 (m, 2 H), 7.53 (d, 1 H), 7.28 - 7.41 (m, 2 H), 7.17 - 7.25 (m, 1 H), 7.04 - 7.14 (m, 1 H), 6.96 - 7.04 (m, 1 H), 5.10 and 5.16 (s, 2 H), 4.98 - 5.15 (m, 1 H), 4.23 - 4.39 (m, 1 H), 3.98 - 4.23 (m, 1 H), 3.46 - 3.85 (m, 5 H), 3.01 - 3.24 (m, 2 H), 1.66 - 2.16 (m, 5 H), 1.26 and 1.36 (s, 9 H);

LC-MS (ESI POS): 532.32 (M+).

EXAMPLE 45

Preparation of ((R)-3-((S)-3-(4-(benzyloxy)phenyl)-2-(tert- butoxycarbonylamino)propanoyloxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane bromide (Diastereomer 1 of C120)

Diastereomer 1 of C119 Diastereomer 1 of C120

Scheme 46

Bz stands for benzyl group.

Preparation of (S)-((R)-quinuclidin-3-yl) 3-(4-(benzyloxy)phenyl)-2- (tert-butoxycarbonylamino)propanoate (Diastereomer 1 of CI 19)

A mixture of (S)-3-(4-(benzyloxy)phenyl)-2-(tert- butoxycarbonylamino)propanoic acid (500 mg, 1.35 mmol), DCC (333 mg, 1.61 mmol) and HOBT (247 mg, 1.61 mmol) in dry THF (10 ml) was stirred at RT under nitrogen atmosphere for 1 hour. (R)-quinuclidin-3-ol (205 mg, 1.61 mmol) was added and the reaction was stirred for 16 hours. The solvent was evaporated and the residue was portioned between sat. NaHCO ₃ and EtOAc. The organic phase was washed with brine, dried over Na ₂ SO ₄ filtered and evaporated to dryness. The residue was triturated with Et ₂ O (30 ml) and the insoluble material was filtered off. The solution was evaporated to obtain (S)-((R)-quinuclidin-3-yl) 3-(4-(benzyloxy)phenyl)-2-(tert- butoxycarbonylamino)propanoate (590 mg, 91% yield).

Preparation of (R)-3-((S)-3-(4-(benzyloxy)phenyl)-2-(tert- butoxycarbonylamino)propanoyloxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane bromide (Diastereomer 1 of C120)

To a solution of (S)-((R)-quinuclidin-3-yl) 3-(4-(benzyloxy)phenyl)-2- (tert-butoxycarbonylamino)propanoate (590 mg, 1.23 mmol) in EtOAc (15 ml), 2-bromo-l-phenylethanone (232 mg, 1.17 mmol) was added portionwise. The reaction was stirred at RT for 16 hours. The solvent was evaporated and the residue was triturated with Et ₂ O, filtered and dried under vacuum to obtain (R)-3-((S)-3-(4-(benzyloxy)phenyl)-2-(tert- butoxycarbonylamino)propanoyloxy)- l-(2-oxo-2-phenylethyl)- 1- azoniabicyclo[2.2.2]octane bromide (720 mg, 86% yield).

1H NMR (300 MHz, DMSO-d6) δ ppm 7.88 - 8.11 (m, 2 H), 7.70 - 7.84 (m, 1 H), 7.56 - 7.69 (m, 2 H), 7.26 - 7.50 (m, 6 H), 7.18 (m, 2 H), 6.94 (m, 2 H), 5.19 (s, 2 H), 5.15 - 5.26 (m, 1 H), 5.07 (s, 2 H), 4.06 - 4.32 (m, 2 H), 3.50 - 3.83 (m, 5 H), 2.79 - 3.09 (m, 2 H), 2.19 (br. s., 1 H), 1.96 - 2.13 (m, 2 H), 1.74 - 1.96 (m, 2 H), 1.35 (s, 9 H);

LC-MS (ESI POS): 599.37 (M+);

[cc] _D = -22.52 (c= 0.5,MeOH).

EXAMPLE 46

Preparation of (R)-3-((S)-2-(tert-butoxycarbonylamino)-3-(4- hydroxyphenyl)propanoyloxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane bromide (Diastereomer 1 of C122)

Diastereomer 1 of C120 Diastereomer 1 of C121 Diastereomer 1 of C122

Scheme 47

Preparation of (S)-((R)-quinuclidin-3-yl) 2-(tert- butoxycarbonylamino)-3-(4-hydroxyphenyl)propanoate (Diastereomer 1 of C121)

A mixture of (R)-3-((S)-3-(4-(benzyloxy)phenyl)-2-(tert- butoxycarbonylamino)propanoyloxy)- 1 -(2-oxo-2-phenylethyl)- 1 - azoniabicyclo[2.2.2]octane bromide (720 mg, 1.06 mmol) and 10% Pd/C (100 mg, 0.09 mmol) in MeOH (30 ml) was hydrogenated at 25 psi for 2 hours. The catalyst was filtered off and the solvent was evaporated to dryness. The residue was dissolved in DCM and filtered through a SCX cartridge. The solution was evaporated to obtain (S)-((R)-quinuclidin-3-yl) 2-(tert- butoxycarbonylamino)-3-(4-hydroxyphenyl)propanoate (260 mg, 62.9% yield).

Preparation of (R)-3-((S)-2-(tert-butoxycarbonylamino)-3-(4- hydroxyphenyl)propanoyloxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane bromide (Diastereomer 1 of C122)

To a solution of (S)-((R)-quinuclidin-3-yl) 2-(tert- butoxycarbonylamino)-3-(4-hydroxyphenyl)propanoate (260 mg, 0.67 mmol) in dry acetonitrile (10 ml), 2-bromo- l-phenylethanone (126 mg, 0.63 mmol) was added portionwise. The reaction was stirred at RT for 3 hours. The solvent was evaporated and the residue was purified by flash chromatography (DCM/MeOH=85/15) and the recovered product was triturated with z-PrOH to obtain (R)-3-((S)-2-(tert-butoxycarbonylamino)-3-(4-hydroxyphenyl)- propanoyloxy)- l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2]octane bromide (74 mg, 18.8% yield).

1H NMR (300 MHz, DMSO-d6) δ ppm 9.22 (s, 1 H), 7.85 - 8.05 (m, 2 H), 7.76 (t, 1 H), 7.62 (t, 2 H), 7.33 (d, 1 H), 7.05 (m, 2 H), 6.69 (m, 2 H), 5.20 (s, 2 H), 4.31 (d, 1 H), 4.01 - 4.25 (m, 2 H), 3.47 - 3.78 (m, 5 H), 2.75 - 3.02 (m, 2 H), 2.20 (br. s., 1 H), 1.76 - 2.13 (m, 4 H), 1.36 (s, 9 H);

LC-MS (ESI POS): 509.33 (M+);

[cc] _D =-15.48 (c=0.5,MeOH). EXAMPLE 47

Preparation of (R)-3-((S)-2-(tert-butoxycarbonylamino)-3- phenylpropanoyloxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane bromide (Diastereomer 1 of C124)

Diastereomer 1 of C123 Diastereomer 1 of C124

Scheme 48

Preparation of (S)-((R)-quinuclidin-3-yl) 2-(tert- butoxycarbonylamino)-3-phenylpropanoate (Diastereomer 1 of C123)

A mixture of (S)-2-(tert-butoxycarbonylamino)-3-phenylpropanoic acid

(500 mg, 1.88 mmol), DCC (467 mg, 2.26 mmol) and HOBT (346 mg, 2.26 mmol) in dry THF (15 ml) was stirred for 30 minutes at RT. Then (R)- quinuclidin-3-ol (288 mg, 2.26 mmol) was added portionwise and the reaction was stirred at RT for 16 hours. The solvent was evaporated, the residue was dissolved in EtOAc and the insoluble was filtered off. The organic phase was washed with sat. NaHCO ₃ and brine, dried over Na ₂ SO ₄ , filtered and evaporated to obtain (S)-((R)-quinuclidin-3-yl) 2-(tert-butoxycarbonylamino)- 3-phenylpropanoate (600 mg, 85% yield).

Preparation of (R)-3-((S)-2-(tert-butoxycarbonylamino)-3- phenylpropanoyloxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane bromide (Diastereomer 1 of C124)

To a solution of (S)-((R)-quinuclidin-3-yl) 2-(tert- butoxycarbonylamino)-3-phenylpropanoate (600 mg, 1.60 mmol) in EtOAc (10 ml), 2-bromo-l-phenylethanone (303 mg, 1.52 mmol) was added portionwise. The reaction was stirred at RT for three days. The precipitate was collected by filtration and dried under vacuum to obtain (R)-3-((S)-2-(tert- butoxycarbonylamino)-3-phenylpropanoyloxy)- l-(2-oxo-2-phenylethyl)- l- azoniabicyclo[2.2.2]octane bromide (740 mg, 81% yield).

1H NMR (300 MHz, DMSO-d6) δ ppm 7.94 - 8.05 (m, 2 H), 7.69 - 7.81

(m, 1 H), 7.54 - 7.69 (m, 2 H), 7.41 (m, 1 H), 7. 17 - 7.36 (m, 5 H), 5.20 (s, 2 H), 5.07 - 5.3 1 (m, 1 H), 4.21 - 4.38 (m, 1 H), 4. 15 (dd, 1 H), 3.48 - 3.86 (m, 5 H), 2.77 - 3. 13 (m, 2 H), 2.15 - 2.25 (m, 1 H), 1.99 - 2. 15 (m, 2 H), 1.75 - 1.99 (m, 2 H), 1.35 (s, 9 H);

LC-MS (ESI POS): 493.45 (M+);

[cc] _D =- 19.72 (C=0.5, MeOH).

EXAMPLE 48

Preparation of (3R)-3-(2-(2-amino-2-phenylacetamido)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo [2.2.2]octane 2,2,2-trifluoroacetate 2,2,2-trifluoroacetic acid (C127)

Scheme 49 Preparation of (R)-quinuclidin-3-yl 2-(2-(tert- butoxycarbonylamino)-2-phenylacetamido)-2-phenylacetate (C125)

To a suspension of (R)-quinuclidin-3-yl 2-amino-2-phenylacetate dihydrochloride (500 mg, 1.50 mmol) in THF (15 ml) and DCM (5 ml), triethylamine (591 μΐ, 4.25 mmol) was added and the reaction was stirred at RT for 10 minutes. Then DCC (309 mg, 1.50 mmol), HOBT (203 mg, 1.50 mmol) and 2-(tert-butoxycarbonylamino)-2-phenylacetic acid (314 mg, 1.25 mmol) were added and the reaction was stirred at RT for 15h. The solvent was evaporated, the residue was taken up with DCM and the insoluble was filtered off. The organic solution was washed twice with IN Na ₂ CO ₃ and brine, dried over Na ₂ SO ₄ and evaporated. The crude was purified by flash chromatography (DCM/MeOH=95/5) to obtain (R)-quinuclidin-3-yl 2-(2-(tert- butoxycarbonylamino)-2-phenylacetamido)-2-phenylacetate (210 mg, 34% yield).

Preparation of (3R)-3-(2-(2-(tert-butoxycarbonylamino)-2- phenylacetamido)-2-phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane chloride (C126)

To a solution of (R)-quinuclidin-3-yl 2-(2-(tert-butoxycarbonylamino)- 2-phenylacetamido)-2-phenylacetate (210 mg, 0.42 mmol) in EtOAc (3 ml) and acetonitrile (3 ml), 2-chloro-l-phenylethanone (65.8 mg, 0.42 mmol) was added and the reaction was stirred at RT for 15h. More 2-chloro- l- phenylethanone (19.7 mg, 0.13 mmol) was added and the reaction was stirred at RT for 24h. The solvent was evaporated and the crude was purified by flash chromatography (DCM/MeOH=94/6 to 93/7) to obtain (3R)-3-(2-(2-(tert- butoxycarbonylamino)-2-phenylacetamido)-2-phenylacetoxy)- l -(2-0X0-2- phenylethyl)-l-azoniabicyclo[2.2.2]octane chloride (65.3 mg, 23.7% yield).

Preparation of (3R)-3-(2-(2-amino-2-phenylacetamido)-2- phenylacetoxy)-l-(2-oxo-2-phenylethyl)-l-azoniabicyclo[2.2.2 ]octane 2,2,2-trifluoroacetate 2,2,2-trifluoroacetic acid (C127)

To a solution cooled at 0°C of (3R)-3-(2-(2-(tert- butoxycarbonylamino)-2-phenylacetamido)-2-phenylacetoxy)- l-(2-oxo-2- phenylethyl)-l-azoniabicyclo[2.2.2]octane chloride (50 mg, 0.08 mmol) in dioxane (4 ml), hydrogen chloride (4M solution in dioxane, 23 μΐ) was added and the reaction was stirred at RT for 15h. Other hydrogen chloride (4M solution in dioxane, 193 μΐ) was added and the reaction was stirred at RT for 7h. Then 37% hydrogen chloride (0.5 ml, 6.09 mmol) was added and the reaction was stirred at RT for 20h. The solvent was evaporated and the resulting crude was purified by preparative HPLC to obtain (3R)-3-(2-(2- amino-2-phenylacetamido)-2-phenylacetoxy)-l-(2-oxo-2-phenyle thyl)-l- azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate 2,2,2-trifluoroacetic acid (14.8 mg, 25.9% yield).

1H NMR (300 MHz, DMSO-d6) δ 8.95 - 9.30 (m, 1 H), 7.86 - 8.1 1 (m, 2 H), 7.69 - 7.86 (m, 1 H), 7.52 - 7.69 (m, 2 H), 7.22 - 7.50 (m, 11 H), 5.30 - 5.57 (m, 2 H), 5.00 - 5.30 (m, 3 H), 4.03 - 4.17 (m, 1 H), 3.43 - 3.78 (m, 5 H), 2.14 - 2.22 and 2.32 - 2.45 (m, 1 H), 1.88 - 2.13 (m, 2 H), 1.71 - 1.88 (m, 1 H), 1.50 - 1.71 (m, 1 H), 1.36 and 1.38 (s, 9 H);

LC-MS (ESI POS): 612.53 (M+).

EXAMPLE 49

Preparation of (3R)-3-(2-benzamido-3-methylbutanoyloxy)-l-(2- oxo-2-phenylethyl)-l-azoniabicyclo [2.2.2] octane 2,2,2-trifluoroacetate (C130

C130

Scheme 50

Preparation of 2-benzamido-3-methylbutanoic acid (C128)

A solution of 2-amino-3-methylbutanoic acid (500 mg, 4.27 mmol) in 2N NaOH (2.35 ml, 4.69 mmol) was stirred at RT for 30 minutes, then the reaction mixture was cooled at 0°C and benzoyl chloride (471 μΐ, 4.05 mmol) and 2N NaOH (2.35 ml, 4.69 mmol) were simultaneously added dropwise from two different syringes. The reaction was stirred at RT for 3h. Water was added, the aqueous phase was washed with Et ₂ O, then acidified with 1M HCl and back-extracted with Et ₂ O, dried over Na ₂ SO ₄ and evaporated. The residue was triturated with z-Pr ₂ O to obtain 2-benzamido-3-methylbutanoic acid (780 mg, 83% yield). Preparation of (R)-quinuclidin-3-yl 2-benzamido-3- methylbutanoate (C129)

To a solution of 2-benzamido-3-methylbutanoic acid (780 mg, 3.53 mmol) in THF (25 ml), (R)-quinuclidin-3-ol (538 mg, 4.23 mmol), DCC (873 mg, 4.23 mmol) and HOBt (572 mg, 4.23 mmol) were added and the reaction was stirred at RT for 15h. The solvent was evaporated. EtOAc was added and the insoluble was filtered off. The organic solution was washed twice with IN Na ₂ CO ₃ and brine, dried over Na ₂ SO ₄ and evaporated to dryness. The crude was purified by flash chromatography (DCM/MeOH =99/1 to 8/2) to obtain (R)-quinuclidin-3-yl 2-benzamido-3-methylbutanoate (436 mg, 37.4% yield).

Preparation of (3R)-3-(2-benzamido-3-methylbutanoyloxy)-l-(2- oxo-2-phenylethyl)-l-azoniabicyclo [2.2.2] octane 2,2,2-trifluoroacetate (C130)

To a solution of (R)-quinuclidin-3-yl 2-benzamido-3-methylbutanoate

(218 mg, 0.66 mmol) in EtOAc (2 ml) and acetonitrile (5 ml), 2-bromo-l- phenylethanone (158 mg, 0.79 mmol) was added and the reaction was stirred at RT for 15h. The solvent was evaporated and the residue was triturated with acetonitrile. The crude was purified by preparative HPLC to obtain (3R)-3-(2- benzamido-3-methylbutanoyloxy)-l-(2-oxo-2-phenylethyl)-l- azoniabicyclo[2.2.2]octane 2,2,2-trifluoroacetate (15.5 mg, 4.2% yield)

1H NMR (300 MHz, DMSO-d6) δ 8.64 and 8.66 (d, 1 H), 7.94 - 8.05 (m, 2 H), 7.83 - 7.94 (m, 2 H), 7.69 - 7.81 (m, 1 H), 7.41 - 7.66 (m, 5 H), 5.22 - 5.32 (m, 1 H), 5.18 and 5.20 (s, 2 H), 4.26 - 4.45 (m, 1 H), 4.07 - 4.24 (m, 1 H), 3.53 - 3.85 (m, 5 H), 1.79 - 2.44 (m, 6 H), 1.06 (d, 3 H), 1.04 (d, 3 H);

LC-MS (ESI POS): 449.24 (M+).

EXAMPLE 50

Preparation of (3R)-3-(2-benzamido-3-methylbutanoyloxy)-l-(2- oxo-2-(thiophen-2-yl)ethyl)-l-azoniabicyclo [2.2.2] octane 2,2,2- trifluoroacet

Scheme 51

To a solution of (R)-quinuclidin-3-yl 2-benzamido-3-methylbutanoate (218 mg, 0.66 mmol) in EtOAc (2 ml) and CH ₃ CN (5 ml), 2-bromo- l- (thiophen-2-yl)ethanone (162 mg, 0.79 mmol) was added and the reaction was stirred at RT for 15h. The solvent was evaporated and the crude was purified by flash chromatography (DCM/MeOH =98/2 to 85/15).

The product was triturated with /-Pr ₂ O and further purified by preparative HPLC to obtain (3R)-3-(2-benzamido-3-methylbutanoyloxy)-l-(2- oxo-2-(thiophen-2-yl)ethyl)-l-azoniabicyclo[2.2.2]octane 2,2,2- trifluoroacetate (30.6 mg, 8.2% yield).

1H NMR (300 MHz, DMSO-d6) δ 8.66 (d, 1 H), 8.21 (dd, 1 H), 8.07 (dd, 1 H), 7.82 - 7.94 (m, 2 H), 7.53 - 7.63 (m, 1 H), 7.43 - 7.53 (m, 2 H), 7.3 1 - 7.39 (m, 1 H), 5.18 - 5.31 (m, 1 H), 4.93 - 5.17 (m, 2 H), 4.32 and 4.39 (t, 1 H), 4.08 - 4.23 (m, 1 H), 3.58 - 3.86 (m, 5 H), 1.72 - 2.43 (m, 6 H), 0.93 - 1.16 (m, 6 H);

LC-MS (ESI POS): 455.26 (M+).

Biological characterisation

EXAMPLE 51

Examples of Radioligand Binding Assay for Cloned Human Muscarinic Receptors and for Human beta Adrenergic Receptors

CHO-K1 clone cells expressing the human Ml-, M2-, M3- receptors (Euroscreen, Swissprot PI 1229, P08172, P20309, Genbank: J02960 respectively) were harvested in Ca ⁺⁺ /Mg ⁺⁺ free phosphate-buffered saline and collected by centrifugation at 1500 rpm for 10 min, at 4°C min. The pellets were resuspended in ice cold buffer A (15 mM Tris-HCl pH 7.4, 2 mM MgCl ₂ , 0.3 mM EDTA, 1 mM EGTA). Cloned cells expressing Ml-, M2-, and M3- receptors were homogenized by a PBI politron (setting 5 for 15 s). The crude membrane fraction was collected by two consecutive centrifugation steps at 40000 g for 20 min at 4°C, separated by a washing step in buffer A.

The pellets obtained from the three cell lines were finally resuspended in buffer C (75 mM Tris HC1 pH 7.4, 12.5mM MgCl ₂ , 0.3 mM EDTA, 1 mM EGTA, 250 mM sucrose) and aliquots were stored at -80°C.

The day of experiment, Ml-, M2-, and M3-receptor frozen membranes were resuspended in buffer D (50 mM Tris-HCl pH 7.4, 2.5 mM MgCl ₂ , 1 mM EDTA). The non selective muscarinic radioligand [3H]-N-methyl scopolamine (Mol. Pharmacol. 45:899-907) was used to label the Ml, M2, and M3 binding sites. Binding experiments were performed in duplicate (ten point concentrations curves) in 96 well plates at radioligand concentration of 0.1-0.3 nM. The non specific binding was determined in the presence of cold N-methyl scopolamine 10 μΜ. Samples (final volume 0.75 mL) were incubated at RT for 120 min for Ml, 60 min for M2 and 90 min for M3 binding assay.

The reaction was terminated by rapid filtration through GF/B Unifilter plates and two washes (0.75 mL) with cold buffer using a Packard Filtermate Harvester. Radioactivity on the filters was measured by a microplate scintillation counter TopCount NXT (Canberra Packard).

In the present assays, Ki values for the tested compounds were determined from the observed IC50 values according to known methods. A lower Ki value indicates that the tested compound has a higher binding affinity for the receptor.

The Ki M2/Ki M3 ratio for representative compounds of general formula (I) in Scheme 1, are comprised between 30 and 157.

The interaction with M3 muscarinic receptors can be estimated by the results of in vitro studies which evaluated the potency of the test compounds and the offset of the inhibitory activity produced after washout of the antagonists in isolated guinea pig trachea and by the in vivo duration of action against acetylcholine-induced bronchospasm in the guinea pig.

EXAMPLE 52

Plasma stability

In order to demonstrate that the compounds are degraded, stability in human plasma at 1 and 5 hours was tested for the compound of the invention. Briefly 10 μΐ of a stock solution 250 μΜ of the compound in acetonitrile were added to 1 ml of human plasma and samples were incubated at 37°C. Plasma (50 μΐ.) was taken after 0, 1 and 5 hours of incubation and added to 140 μΐ of acetonitrile with addition of verapamil as internal standard (250 ng/ml). Samples were analysed by HPLC-MS/MS analysis.

Plasma stability is calculated as percentage remaining after 1 and 5 hours by dividing the peak area at 1 or 5 hours by the area of the peak at time 0.

After 5 hours of incubation, plasma stability values for representative compounds of general formula (I) in Scheme 1 are comprised between 0.6 and 13.1%, indicating that the compounds of the invention are very unstable in human plasma.