COAD is a term encompassing conditions which exhibit, to differing extents, several major progressively developing clinicopathological features, namely in lammatory swelling of airway walls, hypertrophy of submucosal glands, and hyperplasia of epithelial secretory cells leading to hypersecretion of viscous mucous which cannot be cleared effectively, progressive increase in irreversible bronchospasm and decrease in lung elastic recoil. This complex pathway results in progressive loss of lung function, with respiratory impairment, increasing morbidity and, finally, death.

Thus COAD, and also asthma, are diseases of reduced lung function in which antimuscarinic bronchodilators are known to improve airway patency. However, existing agents are non- selective for smooth muscle muscarinic sites in lung and this reduces their effectiveness as bronchodilators and leads to unwanted side effects. Sub-types of muscarinic receptor are now known to exist in the airways (see P. J. Barnes, P. Minette and J. Maclagan, TIPS, 1988, , 412.); Ml receptors are present on sympathetic nerves and parasympathetic ganglia; M2 receptors on pulmonary cholinergic nerves (pre-junctional inhibitory receptors) and M3 receptors are located on smooth muscle (post- junctional receptors). The compounds of the present invention generally have bronchospasmolytic effects at doses which do not significantly affect receptors in other tissues such as brain, heart, gastro-intestinal tract, eye and salivary gland. Furthermore, they generally show selectivity for the lung post- junctional M3 receptors as opposed to the pulmonary pre- junctional M2 receptors and cardiac M2 receptors. Therapeutic action at some other smooth muscle sites may be envisaged. For

example, the compounds are also likely to be useful in treating urinary incontinence.

Thus the present invention provides a compound of the formula:-

or a pharmaceutically acceptable salt thereof, wherein X is either (a) a phenyl group optionally substituted by 1 or 2 substituents each independently selected from halo, CF_, C--C, alkyl, C..-C, alkoxy and hydroxy or (b) a thienyl group; and "Het" is either (a) a 5-membered nitrogen-containing heterocyclic group attached to the adjacent carbon atom either by a carbon or a ring nitrogen atom and which is selected from imidazolyl, pyrazolyl, triazolyl and tetrazolyl, or (b) an oxadiazolyl or thiadiazolyl group attached to the adjacent carbon atom by a carbon atom, or (c) a 6-membered nitrogen-containing heterocyclic group attached to the adjacent carbon atom by a carbon atom and selected from pyridinyl, pyrazinyl, pyrimidinyl and pyridazinyl, "Hec" being optionally substituted by up to 2 substituents each independently selected from halo, CF_, C. -C, alkyl, C -C, alkoxy, hydroxy, amino and azido; and m is 1 or 2.

"Halo" means F, Cl, Br or I. C, and C, alkyl and alkoxy groups can be straight or branched chain.

X is preferably either (a) a phenyl group optionally substituted by 1 or 2 fluoro atoms or (b) a 3-thienyl group. X is most preferably an unsubstituted phenyl group.

"Het" thus includes, for example, lH-imidazol-1-yl, 2- azido-lH-imidazol-1-yl, 2-amino-lH-imidazol-l-yl, 2-methyl-lH-

imidazol-1-yl, 4-methyl-lH-imidazol-l-yl, imidazol-2-yl, l-methylimidazol-2-yl, lH-l,2,3-triazol-l-yl, l-methyl-1,2,3- triazol-5-yl, lH-l,2,4-triazol-l-yl, 3-bromo-5-methyl-1H-1/2,4- triazol-1-yl, 3-bromo-5-ethyl-lH-l,2,4-triazol-l-yl, 3-bromo-5 ^_ propyl-lH-l,2,4-triazol-l-yl, 3-bromo-5-isopropyl-1H-1,2,4- triazol-1-yl, 3-bromo-1-isobutyl-1H-1,2,4-triazol-1-yl, 5-methyl- lH-l,2,4-triazol-l-yl, 5-ethyl-lH-l,2,4-triazol-l-yl, 5-propyl- lH-l,2,4-triazol-l-yl, 5-isopropyl-lH-l,2,4-triazol-l-yl, 5- isobutyl-lH-l,2,4-triazol-l-yl, 3-chloro-lH-l,2,4-triazol-l-yl, lH-l,2,5-triazol-l-yl, lH-pyrazol-1-yl, l-methyl-ρyrazol-5-yl, lH-tetrazol-1-yl, 1-methyl-tetrazol-5-yl, 2-methyl-tetrazol-5-yl, lH-imidazol-4(5)-yl, lH-pyrazol-4-yl, lH-ρyrazol-3(5)-yl, pyridin-2-, 3- or 4-yl, pyrazin-2-yl, pyrimidin-2-yl, pyrimidin- 4-yl, 3-methyl-l,2,4-oxadiazol-5-yl, pyridazin-3-yl or pyridazin- 4-yl.

"Het" is most preferably an imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxadiazolyl, pyridinyl, pyrimidinyl or pyridazinyl group, all said group being optionally substituted by one or two substituents each selected from C. -C, alkyl and halo (preferably chloro or bromo).

When m is 1, "Het" is most preferably a lH-imidazol-1-yl, lH-l,2,4-triazol-l-yl or 5-methyl-lH-l,2,4-triazol-l-yl group

When m is 2, "Het" is most preferably a l-methylimidazol-2- yl group.

Those skilled in the art will appreciate that there are two asymmetric centres in the compounds (I), namely those at the positions identified as 2- and 3'- in figure (1). All diastereoisomers whether separated or not are within the scope of this invention. The preferred esters are however the 3R- quinuclidinyl esters.

When m is 1, the preferred stereochemistry at position 2 is R. When is 2 , the preferred stereochemistry at position 2 is S. Thus the preferred compounds are either (2R, 3R) 3- quinuclidinyl propanoates or are (2S, 3R) 3-quinuclidinyl butanoates, and can be represented as follows:-

The compounds of the formula (I) are preparable by the following routes:-

Route A

The compounds of the formula (I) can be prepared by the reaction of an ester of the formula (II) with formaldehyde in the presence of a strong base such as lithium or potassium diisopropylamide, potassium t-butoxide or sodium hydride. The strong base reacts with the ester (II) to form the carbanion (IIA), and the carbanion then reacts with the formaldehyde. The formaldehyde is generally provided either as formaldehyde gas, or as paraformaldehyde (which breaks down to formaldehyde in solution).

— (ID

The carbanion has the formula:

The preferred techniques are as follows.

In the preferred technique, sodium hydride or potassium t- butoxide, the ester (II) and paraformaldehyde are reacted together in a suitable organic solvent, e.g. dimethylformamide, at about room temperature. The product (I) can then be isolated and purified conventionally.

In an alternative technique, the ester (II) is reacted for a few hours with lithium diisopropylamide in tetrahydrofuran at about -78°C. The reaction mixture is then slowly allowed to warm to room temperature during which time formaldehyde gas, generated e.g. by heating paraformaldehyde, is intermittently passed into the solution. Alternatively, paraformaldehyde is simply added to the solution.

Compounds (I) having R stereochemistry at position 3' are preferred, and these are best obtained by starting with an ester (II) having R stereochemistry at position 3' in formula (II). Likewise the 3S quinuclidinyl esters can be prepared from esters (II) having S stereochemistry at the 3'-position.

It is usually most convenient to start with the 2RS forms of the esters (II) even if the 2R or 2S, rather than 2RS, end products are required. This will result in a mixture of diastereomers of the compounds (I), and, if desired, these can be separated into the 2R and 2S forms by conventional techniques such as fractional crystallisation or chromatography. As stated above, in general, when m is 1, the (2R, 3R), and when m is 2, the (2S, 3R) forms of the compounds (I) are preferred.

The novel esters (II) also form a part of the invention.

The starting materials (II) in which m is 1 are obtainable by conventional techniques, e.g. as follows:-

(a) This route is generally only suitable for preparing intermediates in which "Het" is a 6-membered heterocycle as previously defined.

(XCH ₂ CO) ₂ 0 3-Quinuclidinol _{> XC} H ₂ COO(3-quinuclidinyl)

e

CH ₂ -Het

\/

CH

(II), m = 1

Q is a suitable leaving group, typically Cl or Br.

(b) This route is generally only suitable for preparing intermediates in which "Het" is either a 6-membered heterocycle or a 5-membered heterocycle linked via carbon to the adjacent carbon atom.

0 0

XC0C0 ₂ H (C0C1) ₂ X-C-C- (3-quinuclidinyl)

3-Quinuclidinol

CH -Het/Strong base (e. g . LDA)

(IV) (III)

Compounds (II), m = 1

It is preferred to use (R)-3-quinuclidinol in the above so as to obtain the preferred (R)-stereochemistry at the 3'- position.

and ( c )

CH.

\ //

C=0

(V)

The compounds (V) can be prepared as described in Route B.

Route B

This reaction can be illustrated as follows:-

(V) (I), = 2

The reaction also proceeds via the carbanion (IIA) (m =

2) - see Route A - but it is not necessary to isolate it.

The reaction can be carried out conventionally. The anion of the heterocycle Het-CH_ can be obtained conventionally by reaction of said heterocycle with a base, preferably a strong base such as n-butyllithium or lithium or potassium diisopropylamide.

The preferred technique is to react the heterocycle Het-CH, with n-butyllithium or lithium diisopropylamide in a suitable organic solvent, e.g. tetrahydrofuran, at about -78 C. After a few hours, the quiuclidine derivative (V) in a suitable organic solvent, e.g. tetrahydrofuran, iss added, and the reaction mixture is stirred at -78 C for a half hour or so and then paraformaldehyde is added and the mixture is allowed to warm slowly to room temperature. After a few hours the desired product (I) is recovered from the reaction mixture by conventional techniques.

The starting materials (V) can be obtained conventionally, e.g. as follows:-

(V)

Again it is preferred to use (R) -3-quinuclidinol .

Route C

This reaction can be illustrated as follows:-

(V) (I). m =- 1

The reaction also proceeds via the carbanion (IIA) (m = 1) - see Route A - but it is not necessary to isolate it.

This route is only suitable for preparing the compounds (I) in which m is 1 and "Het" is (i) a 5-membered heterocycle attached to the adjacent carbon atom by a ring nitrogen atom or (ii) a 5-membered heterocycle attached to the adjacent carbon atom by a carbon atom and containing no hydrogen atoms on any of its ring nitrogen atoms (see e.g. Examples 26-28), or (iii) pyridyl.

The reaction can be carried out conventionally. The anion of the heterocycle Het-H can be obtained conventionally by reaction of said heterocycle with a base, preferably a strong base such as sodium hydride, n-butyllithium, or lithium or potassium diisopropylamide or potassium t-butoxide. Pyridine anions are best prepared by reaction of the appropriate bromo- pyridine with n-butyllithium as is known to those skilled in the art. The formaldehyde can be provided as formaldehyde gas, or as paraformaldehyde which breaks down to formaldehyde in solution.

For preparing compounds in which m is 1 and "Het" is attached via a ring-nitrogen atom to the adjacent carbon atom,

-li¬ the preferred technique is to react the quinuclidine derivative of the formula (V), paraformaldehyde, the heterocycle Het-H and sodium hydride together in a suitable organic solvent, e.g.' dimethylformamide, at about room temperature. The product can then be isolated and purified conventionally.

For preparing compounds in which m is 1 and "Het" contains no hydrogen atoms on its ring nitrogen atoms and is attached by a carbon atom to the adjacent carbon atom, the preferred technique is to react the heterocycle Het-H with n-butyllithium in a suitable organic solvent, e.g. tetrahydrofuran, at about -78 C. After a few hours, the quinuclidine derived (V) in a suitable organic solvent, e.g. tetrahydrofuran, is added, and the reaction mixture is stirred at -78 C for a half hour or so and then allowed to warm slowly to about 0 C when paraformaldehyde is added. After a few hours the desired product (I) is recovered from the reaction mixture by conventional techniques.

In instances where tautomerism occurs in the heterocycle, then more than one anion may be generated by the reaction of the heterocycle with the base, thus producing a mixture of products as in Example 5.

Some of the compounds of the formula (I) (m = 1 or 2) can be prepared from other compounds of the formula (I). For example, an azido-substituent on "Het" can be reduced to amino e.g. by catalytic hydrogenation, and a bromo-substituent on "Het" can be reduced to hydrogen again by catalytic hydrogenation. A typical hydrogenation is carried out in ethanol at about 50 psi (344.7 kPa) hydrogen pressure in the presence of palladium-on- carbon at about room temperature.

The selectivity of the compounds as muscarinic receptor antagonists can be measured as follows.

Male guinea pigs are sacrificed and the ileum, trachea, bladder and right atrium are removed and suspended in Krebs solution under a resting tension of 1 g at 30 C aerated with 95% 0„ and 5% C0_. Contractions of the ileum, bladder and trachea are recorded using an isotonic (ileum) or isometric

transducer (bladder and trachea). The frequency of contraction of the spontaneously beating double atria is derived from isometrically recorded contractions.

Dose-response curves to carbachol are determined using a 1- 5 minute contact time for each dose of agonist until the maximum response is achieved. The organ bath is drained and refilled with Krebs solution containing the lowest dose of the test compound. The test compound is allowed to equilibrate with the tissue for 20 minutes and the agonist dose-response curve is repeated until the maximum response is obtained. The organ bath is drained and refilled with Krebs solution containing the second concentration of test compound and the above procedure is repeated. Typically three concentrations of the test compound are evaluated on each tissue.

The negative log of the molar contration (pA„) of the test compound which causes a doubling of the agonsit concentration to produce the original response is determined by Schild analysis (Arunlakshana and Schild (1959), Brit. J. Pharmacol., 14., 48-58). Using the above pharmacological techniques, tissue selectivity for muscarinic receptor antagonists is determined.

Activity against agonist-induced or nerve-evoked bronchoconstriction, gut or bladder contractility in comparison with changes in heart rate is determined in the anaesthetised dog, cat or guinea pig. Oral activity is assessed in the conscious dog determining compound effects on, lung function, heart rate, pupil diameter and gut motility.

Compound affinity for other cholinergic sites is assessed in the mouse after either intravenous or intraperitoneal administration. Thus, the dose which causes a doubling of pupil size is determined as well as the dose which inhibits the salivation and tremor responses to intravenous oxotremorine by 50%.

The selectivity of the compounds for pulmonary post- injunctional as against pre-junctional muscarinic receptors in anaesthetised guinea pigs and cats can be assessed by the following techniques. Acetylcholine released by nerve

stimulation activates post-junctional M3 muscarinic receptors to cause contraction or airway smooth muscle and, in addition, activates pre-junctional autoreceptors which inhibit further transmitter release. Animal studies indicate that these pulmonary pre-junctional muscarinic autoreceptors are of the M2 subtype (Barnes et al, 1989). Non-selective agents like ipratropium bromide will inhibit both sites, resulting, in the case of nerve-mediated responses, in an increase in transmitter release which can overcome the post-junctional receptor blockade. Published literature has shown that ipratropium bromide can actually potentiate vagally-induced bronchoconstriction in anaesthetised guinea pigs (Fryer and Maclagan, Eur. Jou. Pharmacol., 139. 187-191 (1987)). Thus, the effects of the test compounds on pre- and post- junctional muscarinic sites can be determined in vivo by comparing the effect on nerve mediated responses with the effect on responses to exogenously administered acetylcholine.

For example, the compound of Example 1 has been found to antagonise both acetylcholine-induced, and vagally-induced, bronchoconstriction in anaesthetised guinea pigs over the smae does range. This contrasts with opratropium bromide which is significantly less potent against vagally-induced than against acetylcholine-induced bronchoconstriction. Additionally, at doses below 1 ug kg of ipratropium bromide, vagally-induced bronchoconstriction is actually potentiated, confirming its pre- junctional effects.

Similar results were obtained from the compound of Example 1 in the anaesthetised cat. The animals were pretreated with propranolol because high sympathetic tone under chloralose anaesthesia may oppose potentiation of vagus nerve-induced bronchoconstriction. The test results indicate that, in addition to its high potency, the compound of Example 1, in contrast to ipratropium bromide, does not interrupt negative feedback control of transmitter relase in both guinea-pig and cat. This confirms the demonstrated jji vitro selectivity of this compound for M3 as opposed to M2 muscarinic receptors.

As a result of this selectivity for post- as opposed to pre-junctional muscarinic receptors, the compounds of the invention should be more effective bronchodilators in respiratory disease compared to ipratropium bromide.

The acid addition salts of the compounds of formula (I) can be prepared in a conventional manner by treating a solution or suspension of the free base of (I) with about one chemical equivalent of a pharmaceutically acceptable acid. Conventional concentration and recrystallization techniques are employed in isolating the salts. Illustrative of suitable acids are acetic, lactic, succinic, maleic, tartaric, citric, ascorbic, benzoic, cinnamic, fumaric, sulfuric, phosphoric, hydrochloric, hydrobromic, hydroiodic, sulfamic, sulfonic such as methanesulfonic, benzenesulfonic, and related acids.

For treatment of the various conditions described above the compounds of formula (I) may be administered to a subject in need of treatment by a variety of conventional routes of administration, including oral administration, and in an aerosol or dry powder composition for administration by inhalation. The compounds have potential for absorption through the gastro¬ intestinal tract and thus administration by slow release formulations is also possible.

In general, a therapeutically-effective oral dose for the active compounds of formula (I) is likely to range from 0.01 to 1 mg/kg body weight of the subject to be treated, preferably 0.1 to 0.5 mg kg. In practice the physician will determine the actual dosage which will be most suitable for an individual patient and it will vary with the age, weight and response of the particular patient. The above dosages are exemplary of the average case but there can, of course, be individual instances where higher or lower dosage ranges are merited, and such are within the scope of this invention.

Although the compounds of formula (I) can be administered alone, they will generally be administered in admixture with a pharmaceutical carrier selected with regard to the intended route of administration and standard pharmaceutical practice. For

example, oral administration may be in the form of tablets containing such excipients as starch or lactose, in capsules either alone or in admixture with excipients, in aerosol or dry powder inhaler form, or in the form of elixirs or suspensions containing flavouring or colouring agents.

In a further aspect the invention provides a pharmaceutical composition comprising a compound of the formula (I), or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable diluent or carrier.

The invention also includes a compound of the formula (I) or a pharmaceutically acceptable salt thereof, for use as a medicament.

The invention further includes the use of a compound of the formula (I), or of a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of chronic obstructive airways disease or asthma.

The following Examples illustrate the preparation of the compounds of the formula (I):-

EXAMPLE 1 (R) -3-Ouinuclidinyl (R and S) -3-hydroxy-2- (lH-imidazol-l - y lmethyl ) - 2 -phenylpropanoa t e

Sodium hydride (10 mg of an 80% dispersion in oil) was added to a mixture of (R)-3-qι uclidinyl 2-phenylacrylate (see Preparation 1) (257 mg), paraf- -maldehyde (90 mg) and imidazole (100 mg) in dimethylformaraide (5 ml) at room temperature. After 1 hour the mixture was partitioned between 10% aqu' s potassium carbonate and chloroform. The organic layer was dried over magnesium sulphate and evaporated to leave a residue which was purified by chromatography on silica gel, performing a gradient elution using ethyl acetate/ether/diethylamine (50:50:5) plus methanol (5—-^10%) as eluant. Fractions containing the first eluted diastereoisomer were combined, evaporated and treated with ethereal hydrogen chloride to give the title compound as a dihydrochloride, of (S) stereochemistry at the 2-position, as a white foam (80 mg, 37%, based on single isomer); a portion of this was subsequently crystallised as the (S) free base, m.p. 128-130°C:-

Aπalysis %:-

Found: C,67.49; H,7.07; N.11.69;

^C 20 ^H 25 ^N 3°3 ^ret - ^uires: C,67.58; H,7.09; N.11.82.

Fractions containing the second eluted diastereoisomer were also combined and evaporated to give the title

compound, of (R) stereochemistry at the 2-position, as a white solid (50 mg, 28%, based on single isomer) which was

?s recrystallised from acetone, m.p. 156-158 C, [ ( ] +93.8 (c

1% in ethanol) : -

Analysis %:-

Found: C,67.30; H,7.07; N,11.80;

^C 20 ^H 25 ^N 3°3 ^re 1 ^uires: C,67.58; H,7.09; N,11.82.

EXAMPLE 2 (R) -3-OuinucIidinyl (R and S ) -3-hydroxy-2-phenyl-2- ( lH-pyrazol-l - ylmethyl )propanoate

Sodium hydride (19 mg, as an 80% dispersion in oil) was added to a mixture of (R)-3-quinuclidinyl-2-phenylacrylate (see Preparation 1) (500 mg), paraformaldehyde (176 mg) and pyrazole (265 mg) in dimethylformamide (5 ml). After 1 hour the mixture was partitioned between 10% aqueous potassium carbonate and chloroform. The organic layer was dried over magnesium sulphate and evaporated to leave a residue which was purified by chromatography on silica gel using ethyl acetate/ether/ diethylamine/methanol (50:50:5:5) as the eluant. Appropriate fractions were combined and evaporated to give the two title diastereoisomers as yellow oils: the stereochemistry of the isomers was not determined.

Diastereoisomer 1 (first eluted isomer) (120 mg, 35%)

-H NMR (300 MHz, CDC1 ₃ )^= 1.2-2.1 (m, 5H), 2.4-3.0 (m, 5H), 3.2

(m, 1H), 3.8-4.2 (m, 2H), 4.6-5.1 ( , 3H), 6.2 (s, 1H), 7.2-7.6

(m, 7H) ppm.

Mass spectrum: m/e (MH) = 356

Diastereoisomer 2 (second eluted isomer) (100 mg, 29%)

-H NMR (300 MHz, CDC1 ₃ ) 6= 1.2-1.9 (m, 5H), 2.4-2.8 (m, 5H), 3.2

(m, 1H), 3.8-4.2 (m, 2H), 4.6 (d, 1H), 4.8 (m, 1H), 5.0 (d, 1H),

6.2 (s, 1H) 7.2-7.4 (m, 6H), 7.55 (s, 1H) ppm.

Mass spectrum: m/e (MH) = 356

EXAMPLES 3 to 13 The following tabulated Examples of the general formula:-

were obtained by similar methods to that described in Example 2 using (R)-3-quinuclidinyl 2-phenylacrylate (see Preparation 1) and the appropriate heterocycle. Individual experimental variations, and the absolute stereochemistry at position 2, where identified, is indicated in the Table. "Diastereoisomers 1 and 2" refer simply to the order of elution from the column and not to any stereochemistry.

Example No. H-Het Experimental Variations Analytical Data

Reaction time 24 hours, Diastereoisomer 1, (S)

HN ^' chromatography solvent stereochemistry - white solid,

N CHCl- plus m.p. 184-186°C.

-N/

0 5% MeOH +

0 -5% NH, (aq.) Analysis %•-

Found: C,60.56; H,6.41; N,19.53;

^C 18 ^H 23 ^N 5°3 requires: C,60.49; H,6.49; N,19.60,

Diastereoisomer 2. (R) stereochemistry - white solid, m.p. 171-173°C.

Analysis %•-

Found: C,60.38; H,6.44; N,19.22;

^C 18 ^H 23 ^N 5°3 requires: C,60.49; H,6.49; N,19.60.

Example No. H-Het Experimental Variations Analytical Data

1.2.3-triazoles continued Diastereoisomer 1 - yellow oil.

-H-NMR (300 MHz, 1.2-1.7 (m, 4H),

2.0 (m, IH), 2.4-2.8 (m, 5H), 3.2 (m, IH),

4.1 (m, 2H), 4.85 (m, 3H), 5.25 (d, IH), 7.2-7.4 (m, 5H), 7.6 (s, IH) ppm.

Mass spectrum: m/e (M ) = 356

Diastereoisomer 2 - yellow oil.

-H-NMR (300 MHz, = 1.2-1.7 (m, 4H),

2.0 (m, IH), 2.6-2.8 (m, 5H), 3.15 (m, IH),

4.1 (m, 2H), 4.8 (m, 3H), 5.25 (d, IH), 7.1-7.4 (m, 5H), 7.55 (s, IH) ppm.

Mass spectrum: m/e (M ) = 356

Example No. H-Het Experimental Variations Analytical Data

Chromatography solvent Diastereoisomer 1, a mixture of the 4-and CHC1 + 0~* 5%, MeOH and 5-methyl isomers, as a white foam.

HN N 0- H% NH ₃ (aq.)

^H-NMR (300 MHz, CDC1 ₃ )£ = 1.2-2.2 (m, 8H),

Me 2.4-3.0 (m, 5H), 3.2 (m, IH), 3.8-5.0 (m, 5H), 6.0-7.4 (m, 7H) ppm.

Mass spectrum: m/e (MH ) - 370.

Diastereoisomer 2, a mixture of the 4- and 5-methyl isomers, as a white foam.

-H-NMR (300 MHz, CDC1 ₃ )C= 1.2-2.2 (m, 8H), 2.4-2.8 (m, 5H), 3.2 (m, IH), 3.8-4.8 (m, 5H), 6.0-7.4 (m, 7H) ppm.

Mass spectrum: m/e (MH ) = 370.

Example No. H-Het Experimental Variations Analytical Data

Diastereoisomer 1 - (S) stereochemistry as a white solid, m.p. 180 C.

Analysis %:- Found: C,53.28; H,5.60; N,12.17

C. BrN.O. requires: C,53.46; H,5.60;

^• '''reacting centre. N,12.47. 3(5)-Bromo-5(3)- methyl-1,2,4- Diastereoisomer 2 (R) stereochemistry as a triazole was white solid, m.p. 181 C. prepared as described in Analysis %:- Chem.Ber., 2250, Found: C,52.99; H,5.71; N,12.05; 100. 1967. C ₂₀ H ₂₅ BrN ₄ O ₃ requires: C,53.46; H,5.60; N,12.47.

EXAMPLE 14 (R) -3-Ouinclidinyl (R and S) -2- (2-azido- lH- imidazol- l-ylmethyl ) - 3-hydroxy-2-phenylpropanoate

The title compounds, the stereochemistry of which at the 2- position was not characterised, were prepared by a similar method to that described in Example 2 but using 2-azidoimidazole (prepared as described in Tet. Lett., 1523, JJB, 1975).

Diastereoisomer 1 (higher Rf by tic) as a white solid (60%) m.p. 136" (dec).

¹ H-NMR (300 MHz, CDC1 ₃ ) & = 1.2-2.1 (m, 5H), 2.2-3.0 ( , 5H), 3.2

(m, IH), 4.0 (d, IH), 4.25 (d, IH), 4.3 (d, IH), 4.55 (d, IH),

4.95 (m, IH), 6.2 (s, IH), 6.8 (s, IH), 7.1 (m, 2H), 7.35 ( , 3H) ppm.

Mass spectrum: m/e (M ) = 396.

Diastereoisomer 2 (lower Rf by tic) as a brown oil (67%)

¹ H-NMR (300 MHz, CDC1 ₃ ) 5= 1.2-1.8 (m, 4H), 1.95 (s, IH), 2.7 (m, 5H), 3.2 (m, IH), 4.0 (d, IH), 4.2-4.4 (m, 2H), 4.5 (d, IH), 4.95 (m, IH), 6.2 (s, IH), 6.7 (s, IH), 7.05 (m, IH), 7.35 (m, 3H) ppm.

Mass spectrum: m/e (M-N„ ) = 368

EXAMPLE 15 (R)-3-Ouinuclidinyl (R and S)-2-(2-amino-lH-imidazol-l-ylmethyl)- 3-hvdroxy-2-phenylpropanoate

*(R.S)

Diastereomers 1 and 2 of (R)-3-quinuclidinyl (R and S)-2- (2-azido-lH-imidazol-l-ylmethyl)-3-hydroxy-2-ρhenylpropanoa te (see Example 14) (400 mg) were separately hydrogenated in ethanol (15 ml) containing 10% palladium-on-carbon (40 mg) at room temperature in an atmosphere of hydrogen [344.7 kPa, (50 psi)]. Filtration and evaporation gave the two title compounds as amorphous white solids; the stereochemistry at the 2-positions of these diastereomers was not determined.

Diastereoisomer 1 (higher Rf by tic) (350 mg, 93.6%).

¹ H-NMR (300 MHz, CDC1 ₃ ) S = 1.2-1.8 (m, 4H), 2.0 (m, IH), 2.4-2.8 (m, 5H), 3.2 (m, IH), 4.0 (d, IH), 4.2 (m, 2H), 4.7 (d, IH), 4.9 (m, IH), 5.8 (s, IH), 6.4 (s, IH), 7.0-7.4 (m, 5H) ppm.

Mass spectrum: m/e (M ) = 370

Diastereoisomer 2 (lower Rf by tic) (310 mg, 84%)

¹ H-NMR (300 MHz, CDC1 ₃ ) 8 = 1.2-2.1 (m, 5H), 2.6-2.8 (m, 5H), 3.1 (m, IH), 4.0-5.0 (m, 5H), 5.8 (s, IH), 6.4 (s, IH), 7.1-7.4 (m, 5H) ppm.

Mass spectrum: m/e (M ) = 370

EXAMPLE 16 (R)-3-Ouinuclidinvl (R)-3-hvdroxv-2-(5-methvl-lH-l.2.4-triazol-l- ylmethyl)-2-phenylpropanoate

A solution of (R)-3-quinuclidinyl (R)-2-(3-bromo-5-methyl- lH-l,2,4-triazol-l-ylmethyl)-3~hydroxy-2-phenylproρanoate (see Example 9) (1.2 g) in ethanol (25 ml) containing 10% palladium-

on-carbon (120 mg) was stirred for 16 hours under an atmosphere of hydrogen [344.7 kPa (50 psi)] at room temperature, filtered and evaporated to leave a residue that was partitioned between 10% aqueous potassium carbonate and ethyl acetate. The organic layer was dried over magnesium sulphate and the residue, after evaporation, recrystallised from ethyl acetate to leave the title compound as a white solid (0.72 g, 73%), m.p. 194-196"C.

Analysis %:-

Found: C,64.76; H,7.11; N.14.77;

^C 20 ^H 26 ^N 4°3 ^re< l ^uires: C,64.85; H,7.07; N,15.12.

EXAMPLES 17 TO 20 The following tabulated Examples of the general formula:-

were obtained by similar methods to that described in Example 16 by hydrogenation of diastereoisomer 2 of the appropriate bromo- containing quinuclidinyl starting material. Individual experimental variations are indicated in the Table, as are the details of the starting materials. The products by analogy with Example 16 have (R) stereochemistry.

Example No. Het Experimental Variation Analytical Data

18 Purified by chromatography White solid. on silica using CHCl,/MeOH (95:5) as eluant. -H-NMR (300 MHz, CDC1 ) & = 0.9 (t, 3H), 0.95 (m, 2H), 1.2-2.2 (7H), 2.9 (m, 5H), 3.3 (m, IH), 3.4 (B.S., IH), 4.2 (d, IH), 4.3 (d, IH), 4.4 (d, IH), 4.8 (d, IH), 5.0 (m, IH), 7.0 (m, 2H), 7.3 (m, 3H), 7.8 (s,

(See Example 11) IH) ppm.

Mass spectrum: m/e (MH ) = 399.

Example No. Het Experimental Variations Analytical Data

19 Me, Me Purified by chromatography White solid. m.p. 182-183 C. 1 on silica using CHC1, + H-NMR (300 MHz, CDC1 ₃ )5= 0.9 (t, 3H),

—N s MeOH 2 -> 5% as eluant. 1.2 (t, 3H), 1.25-2.0 (5H), 2.5 (m, IH),

„=/ 2.75 (m, 5H), 3.2 (m, IH), 4.25 (m, 2H), 4.5 (d, IH), 4.9 (m, 2H), 7.1 (m, 2H), 7.3 (m, 3H), 7.8 (s, IH) ppm.

Mass spectrum: m/e (MH ) = 399.

(See Example 12)

EXAMPLE 21

(R)-3-Qnin ^. _1C licjJT ^. v1 (R and S ⁾ -3-h _v H _oxy -2- _π hpn _v 1 _. -2-(nyr _a * ₇ i _n - ₉ - vlmethyl)prnpanr.^t-o

(R,S)

To a mixture of (R)-3-quinuclidinyl (R,S)-2-ρhenyl-3- (pyrazin-2-yl)propanoate (see Preparation 5) (300 mg) and paraformaldehyde (60 mg) in dimethylformamide (4 ml) was added sodium hydride (29 mg as an 80% dispersion in oil). The mixture was stirred for 1 hour at room temperature then partitioned between ethyl acetate and 10% aqueous potassium carbonate. The organic layer was dried over magnesium sulphate then evaporated to give a residue which was purified by chromatography on silica gel by gradient elution using chloroform plus methanol (0 -^10%) and aqueous ammonia (0 - 1%). Appropriate fractions were combined and evaporated to give the two title diastereomers, of undefined stereochemistry at the 2-positions, as yellow oils.

Diastereoisomer 1 (higher Rf by tic) (65 mg, 40%)

^L H-NMR (300 MHz, CDC1 ₃ ) % = 1.2-1.7 (m, 4H), 1.95 (m, IH), 2.4- 2.8 (m, 5H), 3.15 (m, 2H), 3.5 (d, 2H), 3.75 (d, 2H), 4.1 (m, 2H), 4.95 (m, 2H), 7.2-7.4 (m, 5H), 8.4 (s, IH), 8.5 (d, IH) ppm.

Mass spectrum: m/e (MH ) = 368

Diastereoisomer 2 (lower Rf by tic) (40 mg, 24.5%)

¹ H-NMR (300 MHz, CDC1 ₃ ) % = 1.2-1.8 (m, 4H), 1.95 (s, IH), 2.4- 2.8 (m, 5H), 3.2 (m, IH), 3.5 (d, IH), 3.8 (d, IH), 4.1 (m, 2H), 4.8 (m, IH), 7.3 (m, 5H), 8.4 (s, IH), 7.5 (s, IH) .

Mass spectrum: m/e (MH ) = 368

-42-

EXAMPLES 22 TO 25

The following tabulated Examples of general formula:-

were obtained by similar methods to that described in Example 21 by hydroxymethylation of the appropriate quinuclidinyl starting material. Individual experimental variations are indicated in the Table, as are the Preparation numbers of the starting materials. Diastereoisomers 1 and 2 refer simply to the order of elution from the column and not to any stereochemistry.

Example No. H-Het Experimental Variations Analytical Data

24 Diastereoisomer 1 - yellow oil.

-H-NMR (300 MHz, CDC1 ₃ )£ = 1.2-2.4 (m, 5H), 2.8-3.2 (m, 5H), 3.2-3.6 (m, 3H), 4.0 (d, IH), 4.2 (d, IH), 5.1 (m, IH), 7.0-7.4 (m, 7H), 8.2 (s, IH), 8.4 (s, IH) ppm.

(See Preparation 11) Mass spectrum: m/e (MH ) = 367

Diastereoisomer 2 - yellow oil.

-H-NMR (300 MHz, CDC1 ₃ ) £ = 1.2-2.2 (m, 5H), 2.6-2.9 (m, 5H), 3.3 (m, IH), 3.5 (m, 2H), 3.9 (d, IH), 4.2 (d, IH), 4.9 (m, IH), 7.0- 7.4 (m, 7H), 8.2 (s, IH), 8.5 (d, IH) ppm.

Mass spectrum: m/e (MH ) = 367

EXAMPLE 26 (R) -3-0uinucl idinyl (R and S ) -3-hvdroxy-2 - ( lH-methylpyrazol - 5-ylmethyl ) -2 -phenylpropanoate

N-butyllithium (2.1 ml of a 1.6 molar solution in hexane) was added to N-methylpyrazole (284 mg) in tetrahydrofuran (10 ml) at -78"C. After 2 hours (R)-3- quinuclidinyl 2-phenylacrylate (see Preparation 1) (771 mg) in tetrahydrofuran (5 ml) was added and the mixture stirred for % hour at -78°C, then allowed to warm slowly to 0 ^* C, when paraformaldehyde (180 mg) was added. After 154 hours the mixture was partitioned between 10% aqueous potassium carbonate and ethyl acetate, the organic layer dried (magnesium sulphate) and evaporated. The residue was purified by chromatography on silica gel by gradient elution using ethyl acetate/ether/diethylamine/methanol (50:50:2.5:2.5 — -50:50:5:5) as eluant. Appropriate fractions were combined and evaporated to give the two title compounds, of undefined stereochemistry at the 2-positions, as white solids.

Diastereoisomer 1 (higher Rf by tic) (100 mg, 18%) m.p. 130- 131°C.

Analygis .%'•-

Found: C,66.87; H,7.26; N,11.10.

^C 21 ^H 27 ^N 3 ⁰ 3* ^%H 2° ^re< l ^uires: C,66.64; H,7.45; N,11.10.

Diastereoisomer 2 (lower Rf by tic) (120 mg, 21%) m.p. 124- 126"C.

Analysis %:-

Found: 0,68.31; H,7.40; N,11.80;

C- ₁ H- ₇ N-0 ₃ requires: C,68.27; H,7.36; N,11.37.

EXAMPLE 27 (R)-3-0uinuclidinyl (R and S)-3-hydroxy-2-(l-methylimidazol- 2-ylmethyl)-2-phenyl propanoate

The title compounds of undefined stereochemistry, were obtained, as yellow oils, by a similar method to that described in Example 26 using N-methylimidazole in place of N-methylpyrazole.

Diastereoisomer 1 (higher Rf by tic) (21%)

^H-NMR (300 MHz, CDC1 ) S = 1.2-1.8 (m, 4H); 2.0 (s, IH), 2.4-2.8 (m, 5H), 3.2 (m, 2H), 3.4 (s, 3H), 3.6 (d, 2H), 4.2

(d, 2H) , 4.4 (d, 2H) , 4.9 (m, IH) , 6. 75 (s , IH) , 6.95 (s , IH) , 7.2-7.4 (m, 5H) ppm.

Mass spectrum: m/e (MH ) = 370

Diastereoisomer 2 (lower Rf by tic) (43%)

^Tt-NMR (300 MHz, CDC1 ₃ ) <S = 1.2-1.8 (m, 4H), 1.95 (s, 1H),

2.6-2.9 (m, 5H), 3.2 (m, 2H), 3.4 (s, 3H), 3.6 (d, 2H), 4.25

(d, IH), 4.4 (d, IH), 4.85 (m, IH), 6.7 (s, IH), 6.95 (s,

IH), 7.2-7.5 (m, 5H) ppm.

Mass spectrum: m/e (MH ) = 370.

EXAMPLE 28 (R)-3-0uinuclidinvl (R and S)-3-hvdroxv-2-(lH-methvl-l .2.3- triazol-5-ylmethyl)-2-phenvlpropanoate

The title compounds, of undefined stereochemistry, were obtained by a similar method to that described in Example 26 using N-methyl-l,2,3-triazole (prepared as described in Bull. Soc. Chim. France, 2998, 1967) in place of N-methylpyrazole.

Diastereoisomer 1 (higher Rf by tic) as a white solid (27%) m.p. 200-205'C.

¹ H-NMR (300 MHz, CDC1 ) 5 = 1.0-1.8 (m, 4H), 2.05 (m, IH) , 2.4-3.0 (m, 5H), 3.2 (m, IH), 3.4 (d, IH), 3.6 (d, IH) , 3.65 (s, 3H), 3.9 (d, IH), 4.25 (d, IH), 4.95 (m, IH), 7.05 (s, IH), 7.15 (m, 2H), 7.4 (m, 3H) ppm.

Mass spectrum: m/e (M ) = 370

Diastereoisomer 2 (lower Rf by tic) as a yellow foam (29%)

^X H-NMR (300 MHz, CDC1 ₃ ) <5 = 1.0-1.8 (m, 4H), 1.95 (m, IH),

2.6-3.0 (m, 5H), 3.2 (m, IH), 3.4 (d, IH), 3.6 (m, 2H), 3.9

(d, IH), 4.3 (d, IH), 5.0 (m, IH), 6.95 (s, IH), 7.05 (m,

2H), 7.3 (m, 3H) ppm.

Mass spectrum: m/e (M ) = 370

EXAMPLE 29 (R)-3-Ouinuclidinyl (R and S)-2-hvdroxvmethyl-4-(l-methyl- imidazol-2-yl)-2-phenylbutanoate

N-Butyllithium (13.6 ml of a 1.6 Molar solution in hexane) was added to 1,2-dimethylimidazole (2.11 g) in tetrahydrofuran (80 ml) at -78 C. After 1 hour the mixture was warmed to -15 C, stirred for % hour and re-cooled to - 78 C when (R)-3-quinuclidinyl 2-phenylacrylate (see Preparation 1) (4.74 g) in tetrahydrofuran (40 ml) was added. After .4 hour paraformaldehyde (1.2 g) was added and the mixture was slowly allowed to reach room temperatur, stirred for 1 hour and partitioned between 10% aqueous sodium carbonate and ethyl acetate. The organic layer was dried over magnesium sulphate and evaporated to give a residue which was purified by chromatography on silica gel using ethyl acetate/ether/diethylamine/methanol (50:50:5.5) as the eluant. Appropriate fractions were combined and evaporated to give the two title compounds, with the C_ stereochemistry indicated, as white solids.

Diastereoisomer 1 (higher Rf by tic), (R) stereochemistry

(0.3 g, 8.4% based on single isomer), m.p. 186-187 C.

Analysis %:-

Found: C,68.18; H,7.44; N,10.76;

^C 22 ^H 29 ^N 3°3 ^rec - ^uires: C,68.90; H,7.62; N,10.96.

Diastereoisomer 2 (lower Rf by tic), (S) stereochemistry

(250 mg, 7.1% based on single isomer), m.p. 197-199 C.

Analysis %:-

Found: C,69.07; H,7.47; N,10.84;

^C 22 ^H 29 ^N 3°3 ^re( - ^uires: C,68.90; H,7.62; N,10.96.

EXAMPLE 30 TO 34 The following tabulated examples of the general formula:-

were obtained by similar methods to that described in Example 29 using (R)-3-quinuclidiπyl 2-phenylacrylate and an appropriate anion (generated from the methylheterocycle of the formula CH -Het and base indicated). Individual experimental variations are as indicated in the tanle, diastereoisomers 1 and 2 merely refer to their relative positioning on tic.

Example No. Het, base Experimental Variations Analytical Data

31 Anion generation totally at Diastereoisomer 1 - white solid, m.p. 130-

-78°C; 132°C. chromatography solvent: n-but oyllithium CHC1 ₃ plus ^H-NMR (300 MHz, CDC1 ₃ )%= 1.2-1.4 (m, 2H),

0 -> 5% MeOH plus 1.5-1.8 (m, 3H), 2.0 (s, IH), 2.5-3.0 (m,

0 -> 0.5% NH- (aq). 8H), 3.2 (m, IH), 4.1 (d, IH), 4.35 (d, IH), 4.9 (m, IH), 7.1 (m, 2H), 7.2-7.4 (m, 5H), 7.6 (m, IH), 8.5 (d, IH) ppm.

Mass spectrum: m/e (MH ) = 381

Diastereoisomer 2 - white solid, m.p. 163-

165°C.

^-NMR (300 MHz, CDC1 ₃ )£= 1.2-1.4 (m, IH), 1.4-1.8 (m, 3H), 1.95 (s, IH), 2.5-2.9 (m, 9H), 3.2 (m, IH), 4.1 (d, IH), 4.35 (d, IH), 4.85 (m, IH), 7.15 (m, 2H), 7.2-7.4 (m, 5H), 7.6 (m, IH), 8.5 (d, IH) ppm.

-I-

Mass spectrum: m/e (MH ) = 381

Example No. Het, base Experimental Variations Analytical Data

32 Anion generation totally at Diastereoisomer 1 - white solid, m.p. 149- -78 C; chromatography 151°C. solvent; EtOAc/Et ₂ 0/ lithium Et ₂ NH/MeOH Analysis %: diisopropylamide (50:50:2.5:2.5) Found: C,69.22; H,7.25; N,10.95

^C 22 ^H 27 ^N 3°3 requires: C,69.27; H,7.14; N,11.02

Diastereoisomer 2 - white solid, m.p. 128-

129°C.

Analysis %: Found: C,69.22; H,7.30; N,10.76 C ₂₂ H ₂₇ N ₃ 0 ₃ requires: C,69.27; H,7.14; N,11.02

Example No, Het, base Experimental Variations Analytical Data

33 Anion generation totally at Diastereoisomer 1 - white solid, m.p. 206-

- -7788°CC;; cchromatography 208°C. solvent: lithium E EttO0AAcc//EEtt ₂ 0/Et ₂ NH/Me0H Analysis %'■ diisopropylamide (50:50:5:10) Found: C,69.00; H,7.08; N,10.83

^C 22 ^H 27 ^N 3°3 requires: C,69.27; H,7.14; N,11.02

Diastereoisomer 2 - yellow oil,

^H-NMR (300 MHz, CDC1 ₃ )£ = 1.2-1.9 (m, 6H), 2.4-2.9 (m, 8H), 3.2 (m, IH), 4.1 (d, IH), 4.3 (d, IH), 4.9 (m, IH), 7.2-7.5 (m, 6H), 9.0 (m, 2H) ppm.

+ Mass spectrum: m/e (MH ) = 382

EXAMPLE 35 (R) -3-0uinuclidinyl (R and S ) -2-hydroxymethyl-2-phenyl-4- (lH-ρyrazol-l-yl)butanoate

(R)-3-Quinuclidinyl (RS)-2-phenyl-4-(lH-pyrazol-1- yDbutanoate (see Preparation 13) (0.3 g) was added to a mixture of paraformaldehyde (40 mg) and potassium tert- butoxide (30 mg) that had been pre-stirred in dimethylformamide (15 ml) for % hour at room temperature. After 1 hour the mixture was partitioned between 10% aqueous sodium carbonate and ethyl acetate. The organic layer was dried over magnesium sulphate and evaporated to give a residue which was purified by chromatography on silica gel performing a gradient elution using ethyl acetate/ether/ diethulamine (50:50:5) plus methanol (5-10%) as the eluant. Appropriate fractions were combined and evaported to give the two title compounds, of undefined stereochemistry, as white solids.

Diastereoisomer 1 (higher Rf by tic) (29 mg, 18% based on single isomer), m.p. 154-156 C.

Analysis %:-

Found: C,68.16; H,7.39; N,11.20;

C„..H-_N_0, requires: C,68.27; H,7.37; N,11.37.

Diastereoisomer 2 (lower Rf by tic) (23 mg, 14.4% based on single isomer), m.p. 138-140 C.

Analysis %:-

Found: C,68.61; H,7.40; N,11.31;

C„ ₁ H _?7 N_0, requires: C,68.27; H,7.37; N,11.37.

EXAMPLE 36 (R)-3-0uinuclidinyl (R and S)-2-hydroxymethyl-2-phenyl-4- (lH.1.2.4-triazol-l-vl)butanoate

The title compounds, of undefined stereochemistry, were obtained as white solids, by a similar method to that described in Example 35 using (R)-3-quinuclidinyl (RS)-2- phenyl-4-(lH,l,2,4-triazol-l-yl)butanoate (see Preparation 14) in place of (R)-3-quinuclidinyl (R,S)-2-phenyl-4-(lH- pyrazol-l-yl)butanote.

Diastereoisomer 1 (higher Rf by tic) (18% based on single isomer) m.p. 182-184 C.

•Analysis %•' - Found: C,64.08; H,6.82; N,14.52

^C 20 ^H 26 ^N 4 ^O 3 ^,κH 2° ^re< - ^uires: C,64.07; H,7.13; N,14.94

Diastereoisomer 2 (lower Rf by tic) (14.6% based on single isomer) m.p. 140-142 C.

-H-NMR (300 MHz, CDC1 ₃ ) , = 1.2-1 .5 (m, IH) , 1 .5-1 .9 (m, 3H) , 2.05 (m, IH) , 2.6-3.1 (m, 7H) , 3.3 (m, IH) , 4.0 (d, IH) , 4.2 (m, 2H) , 4.4 (d, IH) , 4.9 (m, IH) , 7.2-7.5 (m, 5H) , 8.0 (d, 2H) . Mass spectrum: m/e (MH ) = 371.5

EXAMPLE 37 (R) -3-Ouinuclidinyl (R and S ) -2-hydroxymethyl-2-phenyl-4-(2- mgthyi-tetrasQl-5-yl>fruta*noate

Sodium hydride (32 mg, as an 80% dispersion in oil) and paraformaldehyde (0.1 g) in dimethylformamide (5 ml) were stirred at room temperature for X hour. (R)-3- Quinuclidinyl (RS)-2-phenyl-4-(2-methyl-tetrazol-5- yDbutanoate (see Preparation 15) (0.38 g) in dimethylformamide (5 ml) was added, the mixture stirred for £ hour, evaporated, and the residue partitioned between ethyl acetate and water. The organic layer was dried over magnesium sulphate and evaporated to give a residue which was purified by chromatography on silica gel using ethyl acetate/ether/diethylamine/methanol (50:50:5:10) as the eluant. Appropriate fractions were combined and evaporated to give the two title compounds of undefined stereochemistry, as white solids.

Diastereoisomer 1 (higher Rf by tic) (49 mg, 29% based on single isomer) m.p. 172-174 C. Analysis %:-

Found: C,62.55; H,6.73; N,17.86

C ₂₀ H _2? N 0 ₃ requires: C,62.32; H,7.06; N,18.17

Diastereoisomer 2 (lower Rf by tic) (36 mg, 21% based on single isomer) m.p. 168-170 C.

Analysis %:-

Found: C,62.56; H,7.06; N,18.16

C ₂ _H N 0 requires: C,63.32; H,7.06; N,18.17

EXAMPLE 38 (R)-3-0uinuclidinvl (R and S)-2-hvdroxymethyl-2-phenyl-4-(l- methvl-tetrazol-5-vl)butanoate

The title compounds, as white solids of undefined stereochemistry, were obtained by a similar method to that described in Example 37 using (R)-3-quinuclidinyl (RS)-2- phenyl-4-(l-methyl-tetrazol-5-yl)butanoate (see Preparation 16) in place of R-3-quinuclidinyl (RS)-2-phenyl-4-(2-methyl- tetrazol-5-yl)butanoate.

Diastereoisomer 1 (higher Rf by tic) (30% based on single isomer) m.p. 199-200°C.

Analysis %:- Found: C,62.41; H,7.05; N,18.00

^C 20 ^H 27 ^N 5°3 ^re( l ^uires: C,62.32; H,7.06; N,18.17

Diastereoisomer 2 (lower Rf by tic) (23% based on single isomer) m.p. 186-188°C. Analysis %:- Found: C,62.27; H,7.14; N,17.77

^C 20 ^H 27 ^N 5°3 ^rec l ^uires: C,62.32; H,7.06; N,18.17

The following Preparations illustrate the preparation of novel starting materials used in the previous Examples:-

Preparation..1

(R)-3-0uinuclidinyl 2-ρhenylacrylate

Oxalyl chloride (44.2 ml) was added to a solution of 2-phenylacrylic acid (50 g) (prepared as described in J.Chem. Soc, 2557, 123. 1923) and dimethylformamide (fc ml) in chloroform (500 ml). The mixture was stirred for % hour, dimethylformamide (.4 ml) was added and the mixture was stirred for a further % hour, then evaporated to give a residue to which chloroform (2 x 100 ml) was added and then evaporated. The residue was finally dissolved in chloroform (500 ml) and to this solution at 10-15"C was added (R)-3- quinuclidinol (prepared as described in Acta. Pharm. Suec;

281, 16., 1979) dissolved in chloroform (500 ml). The mixture was stirred for % hour, allowed to slowly reach room temperature, evaporated and the residue partitioned between 25% aqueous potassium carbonate and ether. The organic layer was dried over magnesium sulphate, evaporated and the residue recrystallised from hexane to give the title compound as a white solid (66 g, 76%), m.p. 83-85°C.

Analysis %:-

Found: C,74.39; H,7.47; N,5.45;

C _lfi H N0 _£ requires: C,74.67; H,7.44; N,5.44.

Preparation 2 (R)-3-0uinclidinyl 2-phenylglyoxalate

Oxalyl chloride (13.7 ml) was added to a solution of phenylglyoxylic acid (19.7 g) and dimethylformamide (2 drops) in chloroform (160 ml). After 2 hours the solvent was evaporated, the residue dissolved in chloroform (120 ml) and (R)-3- quinuclidinol (20 g) in chloroform (200 ml) was added to this at 0°C. The mixture was stirred at room temperature for 2 hours, washed with 10% aqueous potassium carbonate, then with water, dried over sodium sulphate and evaporated to leave the title compound, as a yellow oil, (27 g, 64%).

-H-NMR (300 MHz, CDCl- _j ) = 1.4-2.0 (m, 4H) , 2.25 (s , IH) , 2.8-3.6 (m, 6H) , 5.2 (m, IH) , 7.2-7.8 (m, 3H) , 8.0-8.2 (m, 2H) ppm.

Preparation 3 (R) -3-Ouinuclidinyl (RS ) -2-hydroxy-2-phenyl-3- (pyrazin-2- yDpropanoate

2-Methylpyrazine (0.94 g) in tetrahydrofuran (THF) (5 ml) was added dropwise to lithium diisopropylamide (LDA) (7.51 ml of a 1.5 molar solution in THF) in THF (20 ml) at - 78 ^* C. After 0.75 hour a solution of (R)-3-quinuclidinyl 2- phenylglyoxalate (see Preparation 2) (2.59 g) in THF (20 ml) was added, the reaction mixture was allowed to reach room temperature, stirred for 1 hour and partitioned between ethyl acetate and 10% aqueous potassium carbonate. The organic layer was then dried over magnesium sulphate and evaporated to leave a resiάu which was purified by chromatography on silica ge-. . arforming a gradient elution using chloroform plus methanol (0 -> 15%). Appropriate fractions were combined and evaporated to give the title compound, as a yellow oil (1.4 g, 39%).

^L H-NMR (300 MHz, CDC1 ) = 1.2-2.0 (m, 5H), 2.6-2.9 (m, 5H), 3.1 (m, IH), 3.4 (d, IH), 3.9 (d, IH), 4.8 (m, IH), 7.35 (m, 3H), 7.65 (m, 2H), 8.4 (m, 2H), 8.55 (s, IH) ppm.

Preparation 4 (R)-3-0uinuclidinyl 2-phenyl-3-(pyrazin-2-vl)acrvlate

Thionyl chloride (0.534 ml) in chloroform (5 ml) was added at O'C to a solution of (R)-3-quinuclidinyl (RS)-2- hydroxy 2-phenyl-3-(pyrazin-2-yl)propanoate (see Preparation 3) (1.3 g) in chloroform (5 ml). After 10 minutes, pyridine (0.6 ml) in chloroform (5 ml) was added and the mixture was stirred for 24 hours, diluted with chloroform, washed with 10% aqueous potassium carbonate, dried over magnesium sulphate and evaporated. The residue was then purified by chromatography on silica gel performing a gradient elution using chloroform plus methanol (0 -> 10%) and ammonia solution (0 -> 1%). Appropriate fractions were combined and evaporated to give the title compound as an off-white solid (0.4 g, 32%) m.p. 125-126°C.

^L H-NMR (300 MHz, CDC1 ) ^ = 1.2-1.8 (m, 4H), 2.2 (s, IH), 2.8 (m, 4H), 3.15 (d, IH), 3.4 (m, IH), 5.2 (m, IH), 7.0 (s, IH), 7.3-7.6 (m, 5H), 8.2 (s, IH), 8.55 (s, IH), 8.6 (s, IH) ppm.

Pre gra ipn 5

(R) -3-0uinuclidinyl (RS) -2-phenyl-3- (ρyrazin-2-yl)proρanoate

A solution of (R)-3-quinuclidinyl 2-phenyl-3-(ρyrazin- 2- yDacrylate (see Preparation 4) (350 mg) in ethanol (20 ml) containing 10% palladium-on-carbon (30 mg) was stirred for 24 hours under an atmosphere of hydrogen [344.7 kPa (50 psi)] at room temperature. The mixture was filtered and evaporated to leave the title compound as an oil (320 mg, 91%).

¹ H-NMR (300 MHz, CDC _j ) ζ = 1.2-2.1 (m, 5H), 2.5-3.0 (m, 5H), 3.2 (m, 2H), 3.7 (m, IH), 4.3 (m, IH), 4.95 (m, IH), 7.2-7.5 (m, 5H), 8.4 (s, 2H), 8.55 (d, IH) ppm.

Preparation 6 (R)-3-0uinuclidinyl (RS ) -2-hydroxy-2-phenyl-3- (ρyrimidin-4- yl)prςφanpate

The title compound, as an oil, (66%) was prepared by a similar method to that described in Preparation 3 using 4- methylpyrimidine in place of 2-methylpyrazine.

^L H-NMR (300 MHz, CDC1 ) = 1.2-2.4 (m, 5H), 2.5-3.0 (m, 5H), 3.15 (m, IH), 3.4 (m, IH), 3.9 (m, IH), 4.8 (m, IH), 7.2-7.7 (m, 6H), 8.6 (m, IH), 9.1 (s, IH) ppm.

Preparation 7 (R)-3-Ouinclidinvl 2-phenvl-3-(pvrimidin-4-yl)acrylate

The title compound, as a brown solid (97%) was prepared by a similar method to that described in Preparation 4 using (R)-3-quinuclidinyl (RS)-2-hydroxy-2- phenyl-3-(pyrimidin-4- yDpropanoate (see Preparation 6) instead of (R)-3-quinuclidinyl (RS)-2-hydroxy-2-phenyl-3- (pyrazin-2-yDpropanoate.

¹ H-NMR (300 MHz, CDC1 ₃ )$ = (m, IH), 3.35 (m, IH), 5.2 (m, IH), 6.8 (s, IH), 7.3 (s, IH), 7.4-7.6 (m, 5H), 8.7 (d, IH), 9.05 (s, IH) ppm.

Preparation 8 (R) -3-0uinuclidinyl (RS ) -2-phenvl-3- (pvrimidin-4- yDprppgnoate

The title compound, as an oil, (95%) was prepared by a similar method to that described in Preparation 4 using (R)- 3-quinuclidinyl-2-phenyl-3-(pyrimidin-4-yl)acrylate (see Preparation 7) instead of (R)-3-quinuclidinyl-2-phenyl-3- (ρyrazin-2-yl)acrylate.

^-NMR (300 MHz, CDCl.^ = 1.6-2.4 (m, 5H), 2.8-3.7 (m, 6H), 4.1 (m, 2H), 4.7 (m, IH), 5.2 (m, IH), 7.5 (m, IH), 7.7 (m, 5H), 8.9 (d, IH), 9.4 (d, IH) ppm.

Mass spectrum: m/e (M ) = 337

Preparation 9 (R)-3-Quinuclidinyl 2-phenylacetate

(PhCH ₂ C0) ₂ 0

Phenylacetic acid anhydride (prepared as described in J. Org. Chem., 1588, 2 , 1965) (15 g) was added to a suspension of (R)-3-quinuclidinol (5 g) in ethyl acetate (250 ml) at room temperature. After hour the solvent was evaporated and the residue dissolved in hydrochloric acid (2M). This was washed with ethyl acetate, basified with sodium carbonate and extracted with ethyl acetate. The organic extract was dried over sodium sulphate and evaporated to give the title compound as a yellow oil (9 g, 66%).

Analysis %:-

Found: C,72.95; H,7.69; N,5.43;

^C 15 ^H 19 ^N0 2 ^re( l ^uires:C . ⁷³ - 4; H,7.81; N,5.71.

Preparation 10 (R)-3-0uinuclidinyl ⁽ RS)-2-phenyl-3-(pyridin-2-yl)proρanoate

A mixture of (R)-3-quinuclidinyl 2-phenylacetate (see Preparation 9) (1.23 g) and sodium hydride (165 mg of an 80% dispersion in oil) in dimethylformamide (10 ml) was stirred for 1/4 hour, treated with 2-ρicolyl chloride (0.64 g), stirred for 24 hours then partitioned between ethyl acetate and 10% aqueous potassium carbonate. The organic layer was dried over magnesium sulphate and the residue, after evaporation, was purified by chromatography on silica gel by gradient elution using chloroform plus methanol (0—10%) and aqueous ammonia (0—H%). Appropriate fractions were

combined and evaporated to give the title compound as an oil (605 mg, 36%).

^L H-NMR (300 MHz, CDC1 ₃ ) J = 1.1-2.0 ( , 5H), 2.2-3.0 (m, 5H), 3.2 (m, 2H), 3.65 (m, IH), 4.3 (m, IH), 4.7 (m, IH), 7.0-7.6 (m, 8H), 8.5 (m, IH) ppm.

Preparation

(R)-3-0uinuclidinyl (RS)-2-phenyl-3-(pyridin-3-yl)propanoate

The title compound, as an oil, (73%) was prepared by a similar method to that described in Preparation 10 using 3- picolyl chloride in place of 2-picolyl chloride.

^L H-NMR (300 MHz, CDC1 ) ^ = 1.2-1.8 (m, 4H), 1.8 (m, IH), 2.5-2.8 (m, 5H), 3.1 (m, IH), 3.25 (m, IH), 3.85 (m, IH), 4.75 (m, IH), 7.2-7.6 (m, 7H), 8.45 (m, 2H) ppm.

Mass spectrum: m/e (M ) = 336

Preparation 12 (R) -3-0uinuclidinyl (RS ) -2-phenyl -3- (pyridin-4-yl )propanoat.

The title compound, as an oil, (25%) was prepared by a similar method to that described in Preparation 10 using 4- picolyl chloride in place of 2-picolyl chloride followed by purification of the crude product by silica gel chromatography eluting with ethyl acetate/ether/diethylamine/methanol (50:50:254:254).

¹ HtNMR (300 MHz, CDC1 ₃ ) = 1.2-1.8 (m, 4H), 1.9 (s, IH), 2.4-2.8 (m, 5H), 3.1 (m, 2H), 3.45 (m, IH), 3.9 (m, IH), 4.7 (m, IH), 7.1 (d, 2H), 7.2-7.5 (m, 5H), 8.5 (d, 2H) ppm.

Mass Spectrum: m/e (M ) = 336

Preparation 13 (R)-3-0uinuclidinyl (RS)-2-ρhenyl-4-(lH-pyrazol-l- yDbutanoate

A mixture of methyl (RS)-2-ρhnyl-4-(lH-pyrazol-l-yl)- butanoate (see Preparation 17) (0.37 g), (R)-3-quinuclidinol (0.24 g) sodium hydride (15 mg, as an 80% dispersion in oil) in toluene (15 ml) was refluxed with continuous removal of distillate amd, when necessary, replacement with frech toluene, for 154 hours. The cooled mixture was successively washed with water then saturated brine and extracted with 2M hydrochloric acid. The aqueous layer was washed with ethyl acetate, basified with potassium carbonate and extracted with ethyl acetate. The organic layer was dried over magnesium sulphate and evaporated to give the title compound (0.31 g, 61%) as a yellow oil.

-H-NMR (300 MHz, CDC1 ₃ )&= 1.0-2.0 (m, 5H), 2.2-2.8 (m, 7H), 3.1 (m, IH), 3.5 (t, IH), 4.1 (m, 2H), 4.8 (m, IH), 6.25 (s,

IH), 7.2-7.4 (m, 6H), 7.55 (s, IH) ppm. Mass spectrum: m/e (M ) = 339

Preparations 14-16 The following tabulated examples of the general formul :-

were obtained by similar methods to that described in Preparation 13 by ester exchange using the appropriately substituted methyl butanoate and (R)-3-quinuclidinol.

Preparation 17 Methyl (RS ) -2-phenyl-4-(lH-pyrazol-l-yl )butanoate

Methyl (RS)-4-chloro-2-phenylbutanoate (prepared as described in J. Amer. Chem. Soc, 443, 7_2., 1951) (1 g) and pyrazole (1 g) were heated together at 120 C for 5 hour, cooled and the residue partitioned between ether and water. The organic layer was washed with water and extracted with 2M hydrochloric acid. The acid extracts were basified with sodium carbonate and extracted with ether. The organic layer was dried over magnesium sulphate and evaported to give the title compound (0.37 g, 33%) as a colourless oil.

^H-NMR (300 MHz, CDC1 ₃ ), = 2.35 (m, IH), 2.7 (m, IH), 3.5 (t, IH), 3.7 (s, 3H), 4.1 (m, 2H), 6.3 (s, IH), 7.2-7.5 (m, 6H), 7.6 (s, IH) ppm.

Preparation 18 Methyl (RS)-2-ρhenyl-4-(lH-l,2.4-triazol-l-yl)butanoate

The title compound, as a colourless oil, was prepared in 31% yield by a similar method to that described in Preparation 17 using 1,2,4-triazole in place of pyrazole.

-H-NMR (300 MHz, CDC1 ₃ ), = 2.2 (m, IH), 2.7 (m, IH), 3.5 (t, IH), 3.7 (s, 3H), 4.15 (m, 2H), 7.2-7.5 (m, 6H), 8.0 (s, IH) ppm.

Preparation 19 Methyl (RS)-2-ρhenyl-4-(2-methyl-tetrazol-5-yl)butanoate and methyl (RS)-2-phenyl-4-(l-methyl-tetrazol-5-vDbutanoate

Methyl iodide (0.34 ml) was added to a mixture of potassium carbonate (1.13 g) and methyl (RS)-2-phenyl-4-(lH- tetrazol-5-yl)butanoate (see Preparation 20) (1.34 g) in acetonitrile (50 ml). After 18 hours the mixture was filtered and the filtrate evaporated to leave a residue which was partitioned between 10% aqueous potassium carbonate and ether. The organic layer was dried over magnesium sulphate and evaporated to leave a residue which was purified by chromatography on silica gel eluting wth ethyl acetate/hexane (40:60). Appropriate fractions wre combined and evaporated to give to two title compounds, as colourless oils.

2-Methyltetrazol-5-yl isomer (higher Rf on tic) (0.38 g, 27%)

-H-NMR (300 MHz, CDC1 ) , ^= 2.3 (m, IH) , 2.55 (m, IH) , 2.85 (m, 2H) , 3.7 (m, 4H) , 4.3 (s , 3H) , 7.3 (m, 5H) ppm.

l-Methyltetrazol-5-yl isomer (lower Rf on tic) (0.51 g, 36%)

-H-NMR (300 MHz, CDC1 ₃ ), > = 2.3 (m, IH), 2.55 (m, IH), 2.8 (m, 2H), 3.65 (s, 3H), 3.75 (t, IH), 3.9 (s, 3H), 7.2-7.5 (m, 5H) ppm.

Preparation 20 Methyl (RS)-2-phenyl-4-(lH-tetrazol-5-yl)butanoate

Methyl (RS) 4-cyano-2-ρhenylbutanoate (see Preparation 21) (1.8 g) and tri-n-butyltin azide (3.23 g) were mixed and heated at 160 C for 3 hours, dissolved in methanol (100 ml), treated by the addition of hydrogen chloride gas for 10 minutes and left for 18 hours. Evaportion gave a residue which was triturated three times with diisopropyl ether then partitioned between 10% aqueous sodium carbonate and ethyl acetate. The aqueous later was acidifed with 2M hydrochloric acid and extracted with ethyl acetate. The

organic layer was dried over magnesium sulphate and evaporated to give the title compound (1.34 g, 61%) as a brown oil.

-H-NMR (300 MHz, CDC1 ), = 2.35 (m, IH), 2.45 (m, IH), 3.05 (t, 2H), 3.65 (s, 3H), 3.75 (t, IH), 7.2-7.5 (m, 5H) ppm.

Preparation 21 Methyl (RS)-4-cvano-2-Phenvlbutanoate

Lithium diisopropylamide (3.67 ml of a 1.5 molar solution in cyclohexane) was added to acetonitrile (0.26 ml) in tetrahydrofuran (10 ml) at -78 C. After 1 hour, methyl 2-ρhenyl-acrylate (see Preparation 22) (0.81 g) in tetrahydrofuran (10 ml) was added and the mixture was stirred for 1 hour, allowed to warm to room temperature then treated with saturated ammonium chloride solution. The resulting mixture was partitioned between ethyl acetate and water, the organic phase dried over magmesium sulphate and evaported to give a residue which was partitioned between ether and 10% aqueous sodium carbonate. The organic layer was dried over magnesium sulphate and evaporated to leave the title compound (0.5 g, 75%) as an oil.

-H-NMR (300 MHz, CDC1 ),S= 2.0-2.4 (m, 4H), 3.7 (s, 3H), 3.8 (t, IH), 7.2-7.5 (m, 5H) ppm. I.R. (thin film) 2220 cm ^"1 (C = N).

Preparation 22

Methyl 2-phenylacrylate

Oxalyl chloride (24 ml) was added to a solution of 2- phenyl-acryclic acid (37 g) and dimethylformamide (0.5 ml) in dichloromethane (400 ml). The ixture was stirred for 1 hour and then evaporated to give a residue to which dichloromethane (50 ml) was added and evaporated. Methanol (200 ml) was added to the residue, which was then stirred for 1 hour and evaporated to give a residue which was partitioned between 10% aqueous sodium bicarbonate and hexane. The organic layer was dried over magnesium sulphate and evaporated to give the title compound as a colourless oil (39-2 g, 97%).

^H-NMR (300 MHz, CDC1 ₃ ),<5= 3.9 (s, 3H), 5.9 (s, IH), 6.4 (s, IH), 7.2-7.5 (m, 5H).