Salts of aza-bicyclic di-aryl ethers and methods to make them or their precursors

Abstract

The present invention relates to salts of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane, to methods for making them or their precursors, to pharmaceutical compositions comprising them, and to their use as medicaments.

Claims

1. A process for the production of a compound of formula I ##STR00013## or a salt thereof, the process comprising: reacting a compound of formula II ##STR00014## or a salt thereof, with a compound of formula V ##STR00015## or a salt thereof, at elevated temperature in the presence of a base and an inert dipolar aprotic solvent, wherein the base is (M.sub.2)OC(R).sub.3, wherein M.sub.2 is sodium or potassium and each R independently is C.sub.1-6alkyl or two R together with the carbon atom they are bound to form C.sub.4-6 cycloalkyl, or the base is a hydroxide base, to form the compound of formula I; and optionally converting the compound of formula Ito a salt thereof.

2. The process according to claim 1, wherein the base is gradually added to the reaction mixture.

3. The process according to claim 1, wherein the base is sodium tert-butanolate or potassium tert-butanolate.

4. The process according to claim 1, wherein the inert dipolar aprotic solvent is dimethylsulfoxide.

5. The process according to claim 4, wherein the reaction mixture further comprises toluene.

6. The process according to claim 1, further comprising producing the compound of formula II or the salt thereof ##STR00016## according to a process comprising: a) reacting a compound of formula III ##STR00017## with a compound of formula IV ##STR00018## and/or with a compound of formula IVA ##STR00019## wherein M.sub.1 is alkali and n is 1, or M.sub.1 is earth alkali and n is 2; in the presence of a palladium catalyst; a base selected from a carbonate base, a phosphate base, a hydroxide base, and an alcoholate base; water and an inert solvent; to form the compound of formula II; and b) optionally converting the compound of formula II to a salt thereof.

7. The process according to claim 6, wherein the reaction of the compound of formula III with the compound of formula IV and/or the compound of formula IVA is carried out at a pH from 10.5 to 13.

8. The process according to claim 6, wherein the inert solvent is a partly water soluble solvent; wherein cysteine is added to the biphasic reaction mixture after formation of the compound of formula II; the phases are separated; and the compound of formula II is isolated from the non-aqueous phase.

Description

REFERENCE EXAMPLE A1: PREPARATION/CHARACTERIZATION OF FREE BASE OF (R)-3-(6-(4-METHYLPHENYL)-PYRIDIN-3-YLOXY)-1-AZA-BICYCLO[2.2.2]OCTANE IN CRYSTALLINE FORM (FORM A)

(1) About 8 mg of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in free base form dissolved in 0.2 ml methanol was dried in vacuum at 40° C. for >5 hours. After drying, acetonitrile was added to the solid residue and the mixture was heated to 40° C. and vortexed for about 2 hours. The mixture was dried and the remaining solid was analyzed by XPRD.

(2) Following this method, a pattern as shown in FIG. 7A (Form A) can be obtained.

(3) Form A of the free base shows low solubility in aqueous media (0.05 mg/ml).

(4) It is hygroscopic: when tested, Loss on drying (LOD) of a sample was 0.1% and moisture gain was 2% at 93% relative humidity (RH).

(5) Its melting point was determined by heating at 2° C./minute to be 106° C. (onset) with subsequent decomposition.

REFERENCE EXAMPLE A2: PREPARATION/CHARACTERIZATION OF FREE BASE OF (R)-3-(6-(4-METHYLPHENYL)-PYRIDIN-3-YLOXY)-1-AZA-BICYCLO[2.2.2]OCTANE IN CRYSTALLINE FORM (FORM B)

(6) About 8 mg of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in free base form dissolved in 0.2 ml methanol was dried in vacuum at 40° C. for >5 hours. After drying, ethanol was added to the solid residue and the mixture was heated to 40° C. and vortexed for about 2 hours. The mixture was dried and the remaining solid was analyzed by XPRD (see FIG. 7B, Form B).

(7) The same experiment was performed using ethanol or isopropanol as solvent. Basically the same XPRD pattern was obtained. As all three solid forms gained an XPRD pattern as described under Reference Example A1 upon further drying, it was concluded that this new form is an alcohol solvate with low association temperature.

EXAMPLE 1: PREPARATION OF MONO-FUMARATE SALT OF (R)-3-(6-(4-METHYLPHENYL)-PYRIDIN-3-YLOXY) 1-AZA-BICYCLO[2.2.2]OCTANE IN CRYSTALLINE FORM

(8) 500 mg of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in free base form were suspended in 20 ml isopropyl alcohol. A stochiometric amount of fumaric acid was added. The resulting solution was stirred at ambient temperature for 14 hours. The precipitate was collected by filtration and analyzed by proton-NMR and XRPD (see FIG. 1). Yield was 85%. Analysis of proton-NMR confirmed salt formation, a base/acid ratio of about 1:1 and the fact that the salt was not a solvate.

Example 1.1: Preparation of mono-fumarate Salt of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in Crystalline Form by Seeded Crystallization

(9) a) Preparation

(10) 7.3 g mono-fumarate of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane (purity>98%; prepared as described e.g. in Example 13.2) was dissolved in ethanol (42.9 g)/isopropanol (8.5 g)/water (7.2 g) at about 50° C., clarified by filtration and added at this temperature gradually over a period of about 8 hours to filtered tertiary-butylmethylether (118.4 g) at a temperature of about 50° C. After about 25% of the filtrate was added, an ultrasonificated suspension of seed crystals of the mono-fumarate of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane (6 mg, prepared e.g. as described in Example 13.2) in isopropanol (0.1 ml) was added to induce crystallization. The product suspension was maintained for another 1 hour at 50° C. and cooled to 0° C. within 8 hours. After another 1 hour at this temperature the solids were isolated by filtration, washed with isopropanol/tertiary-butylmethylether (40 ml, 1:1 mixture) and dried at about 50° C. under reduced pressure to yield the mono-fumarate of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane (5.85 g; 81% of theory; purity>99.5%).

(11) b) Characterization: Particle Size Measurements by Fraunhofer Light Diffraction

(12) Result:

(13) Mono-fumarate salt of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in crystalline form prepared as described according to Example 1.1 was tested. The following values were obtained: x.sub.10=5.6±0.5 μm; x.sub.50=26.8±1.3 μm and x.sub.90=77.3±3.3 μm (N=8).

(14) Procedure:

(15) To about 0.5 g of test substance add some drops of the dispersing aid (1% Octastat 5000 (Octel Corp.) in white spirit (Sangajol, Schweizerhall Chemie)). Mix intensively on a vortex mixer, in order to wet the substance thoroughly and to form a smooth and homogeneous paste. Dilute the paste with white spirit to a final volume of 3-6 ml and mix the dispersion again. Determine the cumulative volume distribution using a laser diffraction instrument, e.g. determine the particle sizes at the undersize values of 10%, 50% and 90% (x.sub.10, x.sub.50, x.sub.90). Measuring device: Sympatec HELOS (Sympatec GmbH; focal length: 500 mm, optical concentration≧5%, duration of measurement: 40 sec).

(16) Dispersion device: Suspension cell (QUIXEL, Sympatec GmbH).

EXAMPLE 2: PREPARATION OF MONO-MALEATE SALT OF (R)-3-(6-(4-METHYLPHENYL)-PYRIDIN-3-YLOXY)-1-AZA-BICYCLO[2.2.2]OCTANE IN CRYSTALLINE FORM

(17) 500 mg of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in free base form were suspended in 5 ml acetonitrile. A stochiometric amount of maleic acid was added. The resulting solution was stirred at ambient temperature for 14 hours. The precipitate was collected by filtration and analyzed by proton-NMR and XRPD (see FIG. 2). Yield was 63%. Analysis of proton-NMR confirmed salt formation, a base/acid ratio of about 1:1 and the fact that the salt was not a solvate.

EXAMPLE 3: PREPARATION OF MONO-HYDROCHLORIDE SALT OF (R)-3-(6-(4-METHYLPHENYL)-PYRIDIN-3-YLOXY)-1-AZA-BICYCLO[2.2.2]OCTANE IN CRYSTALLINE FORM

(18) A 1 L reactor, equipped with a mechanical stirrer, digital thermometer, nitrogen inlet-outlet, reflux condenser and heating mantle was charged with 14.4 g (R)-3-quinuclidinol, 176 g (160 mL) dimethylsulfoxide and 69.5 g (78 mL) 20 wt % potassium tert-butoxide in tetrahydrofuran. The mixture was stirred at 23° C. for 15 minutes and then heated to 95-110° C. over a period of 1 hour to distill off about 40 mL of tetrahydrofuran. Distilling was continued at 110° C. for 30 minutes. The mixture was cooled to 90° C. over a period of 20 minutes. Portionwise, 21 g 5-chloro-2-p-tolylpyridine was added. 11 g (20 mL) dimethylsulfoxide was added. The reaction mixture was heated to 100° C. over a period of 20 minutes and kept at said temperature for 3 hours. The mixture was cooled to 15° C. over a period of 1 hour.

(19) 370 g (500 mL) tert-butyl methyl ether was added. 250 g water was added over a period of 30 minutes, while maintaining the temperature below 25° C. The mixture was stirred for 30 minutes. The layers were separated and a solution of 102 g 20% (v/v) aqueous sodium chloride was added to the organic layer. The mixture was stirred for 15 minutes and the layers were separated. The organic layer was filtered.

(20) A 1 L reactor, equipped with a mechanical stirrer, digital thermometer, addition funnel, nitrogen inlet-outlet, reflux condenser and heating mantle was charged with the above organic layer. 109 g (120 mL) peroxide-free 2-propanol was added. A solution of 16.5 g (17.8 mL) 5.3 N HCl in 2-propanol was added over a period of 40 minutes. The mixture was heated to 53° C. and stirred for 30 minutes. The mixture was cooled to 23° C. over a period of 30 minutes and stirred for 1 hour. The solid was collected by filtration and washed with a solution of 2×37 g (50 mL) of 1% (v/v) peroxide-free 2-propanol/tert-butyl methyl ether. The solid was dried at 55° C. under reduced pressure to afford 20.9 g of the mono-hydrochloride salt of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane. The material was analyzed by XRPD (see FIG. 3).

Example 4.1: Preparation of mono-phosphate Salt of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in Crystalline Form (Form A)

(21) 500 mg of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in free base form were suspended in 20 ml ethanol. A stochiometric amount of phosphoric acid was added. The resulting solution was stirred at ambient temperature for 14 hours. The precipitate was collected by filtration and analyzed by proton-NMR and XRPD (see FIG. 4A, Form A). Yield was 77%. Analysis of proton-NMR confirmed salt formation and the fact that the salt was not a solvate. Elemental analysis was performed confirming a base/acid ratio of about 1:1 (C 57.9% (58.1%), H 6.7% (6.4%), N 7.1% (7.1%) and P 7.7% (7.9%), theoretical values in brackets).

Example 4.2: Preparation of Phosphate Salt of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in Crystalline Form (Form B)

(22) About 2.3 mg of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in free base form was dissolved in 0.15 ml ethanol. A stochiometric amount of phosphoric acid was added. The mixture was dried in vacuum at 40° C. for >5 hours. After drying, 0.1 ml ethanol and 0.05 ml water were added. The mixture was heated to 40° C. and vortexed for about 2 hours. The mixture was dried and the remaining solid was analyzed by XPRD (see FIG. 4B, Form B).

Example 4.3: Preparation of Phosphate Salt of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in Crystalline Form (Form C)

(23) 200 mg of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in free base form was suspended in 2 ml ethanol. One-third of the stochiometric amount of phosphoric acid was added. The slurry obtained was stirred at ambient temperature and held for 14 hours. Solids were separated by filtration and analyzed by XPRD (see FIG. 4C, Form C).

Example 5.1: Preparation of Mono-Succinate Salt of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in Crystalline Form (Form a)

(24) 500 mg of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in free base form were suspended in 5 ml ethanol. A stochiometric amount of succinic acid was added. The resulting solution was stirred at ambient temperature for 14 hours. The precipitate was collected by filtration and analyzed by proton-NMR and XRPD (see FIG. 5, Form A). Yield was 64%. Analysis of proton-NMR confirmed salt formation, a base/acid ratio of about 1:1 and the fact that the salt was not a solvate.

Example 5.2: Preparation of Anhydrous Mono-Succinate Salt of (R)-3-(6-(4-methylphenyl) pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in Crystalline Form

(25) When the Form A of the mono-succinate salt (see Example 5.1) was subjected to heat (under nitrogen) from 25° C. to 115° C., an anhydrous form of mono-succinate salt was observed and analyzed by XRPD (see FIG. 5, Form B).

EXAMPLE 6: PREPARATION OF MONO-MALONATE SALT OF (R)-3-(6-(4-METHYLPHENYL)-PYRIDIN-3-YLOXY) 1-AZA-BICYCLO[2.2.2]OCTANE IN CRYSTALLINE FORM

(26) 500 mg of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane were suspended in 5 ml acetonitrile. A stochiometric amount of malonic acid was added. The resulting solution was stirred at ambient temperature for 14 hours. The precipitate was collected by filtration and analyzed by proton-NMR and XRPD (see FIG. 6). Yield was 77%. Analysis of proton-NMR confirmed salt formation, a base/acid ratio of about 1:1 and the fact that the salt was not a solvate.

EXAMPLE 7: COMPARISON OF PHYSICO-CHEMICAL PARAMETERS OF SALT FORMS

(27) TABLE-US-00011 Aqueous Initial Known solubility water Hygroscopy Decomposition Multiple [mg/ml] content @ 85% RH m.p. temperature forms Free Base 0.05 0.1 ~2 106 224 Yes Fumarate >30 <0.5 0.5 164 207 No Maleate >30 <0.1 0.3 154 207 No Hydrochloride, >30 ~5 ~5 240 262 Yes Form A Phosphate, >30 ~0.5 0.2 222 222 Yes Form A Succinate 2-15 ~4.5 0.3 113 222 Yes Malonate >30 ~0 1.3 140 148 No

EXAMPLE 8: HARD CAPSULES

(28) Hard gelatin capsules, each comprising as active ingredient 0.5, 5 or 25 mg of the mono-fumarate of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane can be prepared as follows:

(29) TABLE-US-00012 % (w/w) % (w/w) % (w/w) for 0.5 mg for 5 mg for 25 mg Ingredient for capsule fill capsules capsules capsules Mono-fumarate of (R)-3-(6-(4- 0.46 4.65 23.23 methylphenyl)-pyridin-3-yloxy)-1- aza-bicyclo[2.2.2]octane Lactose monohydrate 65.24 61.05 42.47 Microcrystalline cellulose 25.00 25.00 25.00 Hypromellose 2.50 2.50 2.50 Sodium croscarmellose 6.00 6.00 6.00 Colloidal silicon dioxide 0.30 0.30 0.30 Magnesium stearate 0.50 0.50 0.50 Purified water* q.s. q.s. q.s. *removed during processing

(30) Preparation process: Mono-fumarate of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane, lactose monohydrate, microcrystalline cellulose, a portion of sodium croscarmellose and hypromellose are dry mixed in a high shear mixer bowl, and granulating fluid (purified water) added. Once the granulation is complete, the wet granules are dried in a fluid bed drier and the dry granules are milled. The remaining sodium croscarmellose and colloidal silicon dioxide are passed through a suitable sieve and added to the dried granular material and blended in a suitable blending shell. This is achieved by co-sieving the sodium croscarmellose and the colloidal silicon dioxide with a portion of the milled granules through a suitable sieve into the blending shell. Similarly, the required amount of sieved magnesium stearate is added to the bulk granule and then mixed in the same blending shell. This final blend is encapsulated into capsules using automated equipment. Weight ratio of capsule fill to empty capsule shells is 2:1.

EXAMPLE 9: TABLETS

Example 9.1: Film-Coated Tablet

(31) Film-coated tablets containing e.g. 0.5 mg of the mono-fumarate of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane may be prepared as follows:

(32) Preparation of Pre-Mix:

(33) Weigh-in mono-fumarate of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane (e.g. approx. 0.7%) and maize starch (e.g. approx. 13%), mix in a tumble blender (approx 100-300 rotations), pass through a sieve of approx. 0.25-1.0 mm mesh-size. Mix in a tumble blender (approx. 100-300 rotations).

(34) Preparation of Final Blend:

(35) To above pre-mix add microcrystalline cellulose (e.g. approx. 25%), sprayed lactose (e.g. approx. 68%), sodium-carboxymethylcellulose XL (e.g. approx. 2%) and Aerosil (e.g. approx. 0.5%) and mix in a tumble blender (approx. 100-300 rotations). Pass this mixture through a sieve of approx. 0.5-1.0 mm mesh-size and mix again (approx. 100-300 rotations). Add the sodium-stearyl-fumarate (e.g. approx. 1.5%) through a handsieve at approx. 0.5-1.0 mm mesh-size and mix in a tumble blender (approx. 30-150 rotations).

(36) Compression:

(37) On a rotary press compress the above final blend to cores of approx. 100 mg, using the dosage specific tooling (e.g. approx. 6 mm, round, curved).

(38) Coating:

(39) Prepare a suspension in water with basic coating premixes black, red, yellow and/or white. Coat the above obtained cores in a perforated coating pan, and dry.

Example 9.2: Bilayer Film-Coated Tablet

(40) Bilayer film-coated tablets containing e.g. 2.5 mg of the mono-fumarate of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane may be prepared as follows:

(41) Final Active Blend:

(42) Weigh-in mono-fumarate of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane coarse (e.g. approx. 15.5%), microcrystalline cellulose (e.g. approx. 25%), sprayed lactose (e.g. approx. 53%), sodium-carboxymethylcellulose XL (e.g. approx. 3%) and Aerosil (e.g. approx. 0.5%) and mix in a tumble blender (approx 100-300 rotations). Pass this mixture through a sieve of approx. 0.5-1.0 mm mesh-size and mix again (approx 100-300 rotations).

(43) Add the Na-stearyl-fumarate (e.g. approx. 3%) through a handsieve at approx. 0.5-10 mm and mix in a tumble blender (approx 30-150 rotations).

(44) Final Placebo Blend:

(45) Weigh-in microcrystalline cellulose (e.g. approx. 26%), sprayed lactose (e.g. approx. 69%), sodium-carboxymethylcellulose XL (e.g. approx. 1.9%) and Aerosil (e.g. approx. 0.5%) and mix in a tumble blender (approx 100-300 rotations). Pass this mixture through a sieve of approx. 0.5-1.0 mm mesh-size and mix again (approx 100-300 rotations).

(46) Add the sodium-stearyl-fumarate (e.g. approx. 3%) through a handsieve at approx. 0.5-1.0 mm and mix in a tumble blender (approx 30-150 rotations).

(47) Compression:

(48) On a rotary press compress the above final blends to a bilayer tablet-core of approx. 100 mg with one placebo layer (approx. 77.5 mg) and one active layer (approx. 22.5 mg), using the dosage specific tooling (e.g. approx. 6 mm, round, curved).

(49) Coating:

(50) Prepare a suspension in water with basic coating premixes black, red, yellow and/or white. Coat the above obtained cores in a perforated coating pan, and dry.

Example 9.3: Film-Coated Tablet

(51) Film-coated tablets containing e.g. 50 mg of the mono-fumarate of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane may be prepared as follows:

(52) Final Blend:

(53) Weigh-in mono-fumarate of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane coarse (e.g. approx. 15.5%), microcrystalline cellulose (e.g. approx. 25%), sprayed lactose (e.g. approx. 53%), sodium-carboxymethylcellulose XL (e.g. approx. 3%) and Aerosil (e.g. approx. 0.5%) and mix in a tumble blender (approx. 100-300 rotations). Pass this mixture through a sieve of approx. 0.5-1.0 mm mesh-size and mix again (approx. 100-300 rotations).

(54) Add the sodium-stearyl-fumarate (e.g. approx. 3%) through a handsieve at approx. 0.5-10 mm and mix in a tumble blender (approx. 30-150 rotations).

(55) Compression:

(56) Compress the above final blend on a rotary press to cores, using the dosage specific tooling (e.g. approx. 15*5.9 mm, round, curved).

(57) Coating:

(58) Prepare a suspension in water with basic coating premixes black, red, yellow and/or white. Coat the above obtained cores in a perforated coating pan, and dry.

EXAMPLE 10: BIOLOGICAL DATA

(59) The usefulness of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in its various forms, e.g. in free base form (Compound A) or in mono-fumarate salt form (Compound B) in the treatment of the above-mentioned disorders can be confirmed in a range of standard tests including those indicated below.

10.1: In-Vitro Tests: Selectivity of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane Against α4β2-Nachr

(60) Based on the activity/selectivity data shown below it is concluded that said compound is a selective agonist at the α7-nAChR.

(61) TABLE-US-00013 α7-nAChR activity Efficacy α4β2-nAChR Potency compared to activity EC.sub.50 epibatidine IC.sub.50 fold Compound (nM) (100%) (nM) EC.sub.50 (nM) selectivity A 35 75 5598 >100′000 164
Assay:

(62) To assess α7-nAChR activity, a functional assay was employed using GH3 cells that recombinantly expressed human α7-nAChR. 40000 cells per well were seeded 48 h prior to the experiment on black 96-well plates (Costar) and incubated at 37° C. in a humidified atmosphere (5% CO.sub.2/95% air). On the day of the experiment, medium was removed by flicking the plates and replaced with 0.1 ml growth medium containing 0.002 mM Fluo-4, (Molecular Probes) in the presence of 2.5 mM probenecid (Sigma). The cells were incubated at 37° C. in a humidified atmosphere (5% CO.sub.2/95% air) for 1 h. Plates were flicked to remove excess of Fluo-4, washed twice with Hepes-buffered salt solution (HBSS, in mM: NaCl 130, KCl 5.4, CaCl.sub.2 2, MgSO.sub.4 0.8, NaH.sub.2PO.sub.4 0.9, glucose 25, Hepes 20, pH 7.4; HBS) and refilled with 0.1 ml of HBS containing antagonist when appropriate. The incubation in the presence of the antagonist lasted 3-5 minutes. Plates were placed in the cell plate stage of a FLIPR device (fluorimetric imaging plate reader, Molecular Devices, Sunnyvale, Calif., USA). After recording of the baseline (laser: excitation 488 nm at 1 W, CCD camera opening of 0.4 seconds) the agonists (0.05 ml) were added to the cell plate using the FLIPR 96-tip pipettor while simultaneously recording the fluorescence. Calcium kinetic data were normalized to the maximal fitted response induced by epibatidine, which is a full agonist at α7-nAChR. Four parameter Hill equations were fitted to the concentration-response. Values of Emax (maximal effect in % compared to the epibatidine response) and EC.sub.50 (concentration producing half the maximal effect in μM) were derived from this fit.

(63) Assay described in: D Feuerbach et al, Neuropharmacology (2005), 48, 215-227. To assess the activity of the compound of the invention on the human neuronal nAChR α4β2, a similar functional assay is carried out using a human epithelial cell line stably expressing the human α4β2 subtype (Michelmore et al., Naunyn-Schmiedeberg's Arch. Pharmacol. (2002) 366, 235).

10.2: IN-VIVO PRECLINICAL TESTS

10.2.1: Oral Bioavailability and Brain Penetration of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in Mice

(64) Based on the pharmacokinetic data shown below it is concluded that the brain concentration of said compound in mice is beyond (or at least equal) to the compound's EC.sub.50 at the α7-nAChR for at least 4 hours following an acute oral dose of 30 μmol/kg of the compound in free base form.

(65) Compound A:

(66) TABLE-US-00014 Brain Ratio Time Plasma (pmoles/ Brain/ Administration (hour) (pmoles/ml ± SD) g ± SD) plasma 30 μmol/kg p.o. 0.25 573 ± 234  4631 ± 1717 8 30 μmol/kg p.o. 0.5 559 ± 143 11430 ± 3441 20 30 μmol/kg p.o. 1 322 ± 135 14948 ± 4716 46 30 μmol/kg p.o. 4 20 ± 16 1272 ± 715 62 30 μmol/kg p.o. 8 3.4 ± 0.8  58 ± 27 17 30 μmol/kg p.o. 24 — — —
Assay:

(67) Compounds were orally (30 μmol/kg) administered. Male mice (30-35 g, OF1/ICstrain) were sacrificed at indicated time points after oral administration. Trunk-blood was collected in EDTA-containing tubes and the brain was removed and immediately frozen on dry ice. To 100 μl plasma 10 μl internal standard (1.0 μmol of a compound with solubility and ionization properties similar to test compounds) was added and extracted three times with 500 μl dichloromethane. The combined extracts were then dried under a stream of nitrogen and re-dissolved in 100 μl acetonitrile/water (70% acetonitrile). Brains were weighed and homogenized in water (1:5 w/v). Two 100 μl aliquots of each homogenate+10 μl of internal standard (same standard as used for the plasma samples) were extracted three times with 500 μl dichloromethane and further processed as the plasma samples. Samples were separated on Beckmann high-performance liquid chromatography equipment system with an autosampler (Gilson 233XL). A 10 min linear gradient (10-70%) of acetonitrile containing 0.5% (v/v) formic acid was used to elute the compounds from Nucleosil CC-125/2 C18 reversed phase (Machery & Nagel) column.

(68) The limit of detection (LOD), defined as the lowest concentration of the extracted standard sample with a signal to noise ratio of ˜3.

10.2.2: Functional Read-Out of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in Mice (Social Recognition Test)

(69) Based on the functional in-vivo data shown below it is concluded that oral dosing of said compound at relevant concentrations leads to a specific effect associated with α7-nAChR (i.e. cognition enhancement in the Social Recognition Test in mouse).

(70) TABLE-US-00015 Reduction in time scrutinizing in % ± Dose Compound SEM at 24 h in mg/kg A 36 ± 6 0.3
Assay:

(71) Social interactions between two experimental animals are influenced by their degree of familiarity: the better they know each other, the less time they spend on mutual scrutiny at each meeting. In agreement with published data in rats (Mondadori et al., 1993) we have observed (i) that an adult mouse shows a shortened scrutiny of a young conspecific if the two mice are brought together again within a short time interval (e.g. 1 hour), (ii) that this curtailment is attributable to memory processes: it does not occur if the familiar young partner is replaced by a strange (unfamiliar) young mouse on the second occasion and (iii) that the adult mouse's recollection of the previously scrutinized juvenile partner fades with the elapsed time, i.e., after 24 h, scrutiny takes just about as long as at the first encounter. Memory enhancing agents (i.e. oxiracetam) facilitate learning to the extent that the previously met (familiar) partner is still remembered after 24 h, whereas in vehicle treated control animals the memory usually fades after less than 1 hour (Thor and Holloway, 1982) or after 2-3 hours.

(72) Baseline-test: Pairs consisting of one adult and one young mouse were assigned at random to the experimental and control groups. In each pair only the adult mouse was orally treated 1 hour before the trial with either vehicle or the test compound. The duration of active contacts of the adult mouse with the young mouse was manually recorded over a period of 3 min, including the following behavioural, approach-related items: sniffing, nosing, grooming, licking, pawing and playing, anogenital exploration and orientation toward the young mouse; orientation, thereby, was defined as tip of nose of the adult mouse less than approximately 1 cm distant from the young mouse's body.

(73) Re-test: Twenty-four hours after the baseline-test, the adults in each treatment group were confronted again with the previously encountered (familiar) partner, whereas the half of the adult animals were put together with the previously encountered (familiar) partner and the other half with another (unfamiliar) young mouse. Again the duration of active approach-behaviours was recorded during a 3-min period. Prior to re-test no oral injection was given. In the table the reduction in time scrutinizing the familiar partner at time 24 compared with the familiar partner at time 0 minutes is given (value of zero would signify no reduction).

10.2.3: Oral Bioavailability of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in Dogs

(74) Based on the pharmacokinetic data shown below it is concluded that the compound reaches significant blood levels in dogs following an acute oral dose of 1.4 μmol/kg of the compound in fumarate salt form.

(75) Compound B:

(76) TABLE-US-00016 Time Plasma Administration (hour) (pmoles/ml ± SD) 1.4 μmol/kg p.o. 0.25 14.7 ± 1.1  1.4 μmol/kg p.o. 0.5 49.4 ± 21.9 1.4 μmol/kg p.o. 1 67.6 ± 22.6 1.4 μmol/kg p.o. 2 75.2 ± 36.8 1.4 μmol/kg p.o. 4 27.5 ± 13.3 1.4 μmol/kg p.o. 8 8.1 ± 3.0 1.4 μmol/kg p.o. 24 0.8 ± 0.7
Assay:

(77) The compound was given to N=3 male beagle dogs in tritiated form:

(78) ##STR00012##

(79) The concentration of the compound in blood was determined by LC-RID. The procedure involved the addition of 5 μg of the compound as internal standard (200 μL of solution containing 25 μg/mL of the compound) to 1 mL of blood. After further addition of 1 mL of water, 0.1 mL of buffer pH 9 and 4 mL of tert-butylmethylether, the samples were shaken for 30 min and centrifuged (4000 g for 10 min at 22° C.). The organic phase was transferred into a tube and evaporated in a Speedvac. The residue was reconstituted in 250 μL of mobile phase—water (80:20 v/v) followed by 75 μL of n-hexane and transferred into an autosampler vial. After centrifugation (13,000 g for 2 min at 22° C.), the hexane layer was pipetted off and discarded, 200 μL of the remainder was injected onto an RP18 column (Waters XTerra, 5 μm, 3.9×150 mm at 40° C.) to separate the compound from potential metabolites and endogenous compounds. The mobile phase of ammonium acetate (10 mM: 0.1% v/v TFA-acetonitrile, 58:42 v/v) was used at a flow rate of 1.0 mL/min. The effluent was monitored by a UV-detector set at 261 nm. The peak corresponding to the unchanged compound was collected in a polyethylene vial by a fraction collector (SuperFrac, Pharmacia LKB) and analyzed for radioactivity. The concentration of the compound in each sample was calculated from the ratio of the amount of radioactivity in the eluate fraction to the area of the ultraviolet absorbance of the non-radiolabeled the compound, that was used as the internal standard.

Example 10.2.4: Pharmacokinetics of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in Rats

(80) Please note that dosing information in this Example 10.2.4 is given relative to the free form, (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane, and is independent from the salt forming agent. If a salt forming agent is used (e.g. fumarate), a corresponding higher amount of the salt will be used to achieve the intended dosing.

(81) Based on the pharmacokinetic data shown below:

(82) After acute oral dosing of 10 mg/kg (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in mono-fumarate salt form, a Cmax of the free form in plasma of 161±53 ng/ml was reached at 0.25-0.5 h. The AUClast amounted to 249±42 h.Math.ng/ml. After acute oral dosing of 10 mg/kg (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in free form, a Cmax of the free form in plasma of 112±40 ng/ml was reached at 0.25-0.5 h. The AUClast amounted to 200±62 h.Math.ng/ml.

(83) Pharmacokinetics of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in Wistar rats after oral administration of 10 mg/kg of the compound in mono-fumarate salt form (Compound B) and in free base form (Compound A) were assessed.

(84) Both compounds were dissolved in Klucel™ (0.5% in water) and were orally administered by gavage (5 ml/kg) to conscious rats (n=8, cross-over design).

(85) K3-EDTA blood (˜0.2 ml) was collected by puncture of a sublingual vein under light isoflurane anesthesia at the following time points: 0.25, 0.5, 1, 2, 3, 4, 6, 8, 24 h post dose. Immediately after sampling, blood samples were put on ice and were processed to plasma by centrifugation at 4° C. within 15 min after sampling (1000 g, 10 min, 4° C.) to obtain ˜100 μl plasma/sample. All plasma samples were stored at −80° C. until analysis. Since the PK study was performed in a cross-over design, a recovery period of approximately 2 weeks took place before the next treatment/blood sampling was performed.

(86) The determination of compounds in plasma was done by LC-MS/MS using electrospray ionization.

a. Concentrations (ng/ml) and derived pharmacokinetic parameters of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in Plasma of Male Wistar Rats Following Oral Administration of 10 mg/kg Free Base

(87) TABLE-US-00017 Phase 1 1 1 1 2 2 2 2 Rat Rat Rat Rat Rat Rat Rat Rat Mean SD CV Time (h) 01 02 03 04 05 06 07 08 All All (%)  0.25 58.8 89.6 83.0 79.4 105    188 49.0 145 100 46.2 46.3  0.5 34.8 51.2 64.5 94.8 119    122 117 65.0 83.5 34.0 40.7  1 20.1 31.4 63.9 36.0 65.8  83.6 87.0 33.8 52.7 25.6 48.5  2 6.79 12.6 51.7 16.2 43.2  46.9 42.2 29.9 31.2 17.3 55.5  3 5.31 8.38 26.3 7.84 23.0  34.4 18.4 6.20 16.2 10.9 67.3  4 3.94 3.99 18.2 4.54 8.63 21.1 13.8 8.55 10.3 6.66 64.4  6 4.87 1.81 6.62 1.27 5.32 4.38 4.59 6.06 4.37 1.90 43.5  8 6.62 0.812 2.46 1.32 3.67 1.54 2.25 9.67 3.54 3.08 86.9 24 1.40 2.44 0.547 1.02 0*   0.632 0.248 0.287 0.822 0.792 96.4 Body weight (kg).sup.b 0.297 0.277 0.288 0.271  0.309 0.318 0.315 0.350 — — — Dose (mg/kg) 10.1 10.2 9.76 10.1 9.88 9.96 10.0 9.65 10.0 0.184 1.85 Tmax (h) 0.25 0.25 0.25 0.50 0.50 0.25 0.50 0.25 0.25 [0.25-0.50].sup.a Tlast (h) 24.0 24.0 24.0 24.0 8.0  24.0 24.0 24.0 24.0  [8.0-24.0].sup.a Cmax (ng/mL) 58.8 89.6 83.0 94.8 119    188 117 145 112 40.4 36.1 Cmax/Dose 5.81 8.79 8.50 9.40 12.0  18.9 11.7 15.0 11.3 4.14 36.7 (ng/mL)/(mg/kg) AUClast (h .Math. ng/mL) 141 123 238 136 214    296 234 236 200 61.9 30.6 AUClast/Dose 14.0 12.0 24.4 13.5 21.6  29.7 23.4 24.5 20.4 6.43 31.5 (h .Math. ng/mL)/(mg/kg) T½ (h) 8.67 nd 5.71 nd 3.24 nd 4.54 3.67 5.17 2.17 42.1 T½ range (h) [6-24] nd [6-24] nd [4-8] nd [6-24] [6-24] .sup.aMedian [range] .sup.bAt time of treatment nd: Not determined due to r.sup.2 <0.75 or AUC % extrapolated >30%. *Values BLOQ (0.150 ng/mL) were set to 0 for Phoenix PK calculations.

b. Concentrations (ng/ml) and derived pharmacokinetic parameters of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in Plasma of Male Wistar Rats Following Oral Administration of 10 mg/kg Mono-Fumarate

(88) TABLE-US-00018 Phase 2 2 2 2 1 1 1 1 Rat Rat Rat Rat Rat Rat Rat Rat Mean SD CV Time (h) 01 02 03 04 05 06 07 08 All All (%)  0.25 167    162 77.8 89.0 242    204 93.3 153 149 58.5 39.4  0.5 104    130 78.4 99.7 143    179 184 138 132 37.4 28.3  1 67.1  88.8 41.3 62.8 83.2  80.8 101 94.2 77.4 19.4 25.1  2 27.4  39.2 37.8 24.6 47.1  37.6 54.8 42.6 38.9 9.81 25.2  3 12.4  17.7 25.3 11.7 19.6  20.6 25.1 23.2 19.5 5.27 27.1  4 9.06 10.6 13.3 5.90 9.71 11.8 13.0 11.1 10.6 2.39 22.6  6 7.49 3.47 7.63 5.79 4.29 4.24 4.05 3.87 5.10 1.66 32.5  8 2.91 2.35 3.62 2.62 2.46 1.94 2.38 2.07 2.54 0.529 20.8 24 0*   0.276 0.210 0.169 0*   1.40 0.162 0.344 0.320 0.453 141 Body weight (kg).sup.b  0.341 0.315 0.330 0.300  0.268 0.266 0.276 0.290 — — — Dose (mg/kg) 9.73 9.86 9.95 9.82 9.61 9.69 10.1 9.77 9.81 0.146 1.49 Tmax (h) 0.25 0.25 0.50 0.50 0.25 0.25 0.50 0.25 0.25 [0.25-0.50].sup.a Tlast (h) 8.0  24.0 24.0 24.0 8.0  24.0 24.0 24.0 24.0  [8.0-24.0].sup.a Cmax (ng/mL) 167    162 78.4 99.7 242    204 184 153 161 52.9 32.8 Cmax/D 17.2  16.4 7.88 10.2 25.2  21.1 18.3 15.7 16.5 5.54 33.6 (ng/mL)/(mg/kg) AUClast (h .Math. ng/mL) 202    259 212 188 269    292 298 272 249 42.2 16.9 AUClast/D 20.8  26.3 21.4 19.2 28.0  30.1 29.6 27.9 25.4 4.30 16.9 (h .Math. ng/mL)/(mg/kg) T½ (h) 2.44 5.02 3.62 3.71 2.02 nd 3.97 5.51 3.76 1.26 40.3 T½ range (h) [4-8] [6-24] [6-24] [6-24] [4-8] nd [6-24] [6-24] .sup.aMedian [range] .sup.bAt time of treatment nd: Not determined due to r.sup.2 <0.75. *Values BLOQ (0.150 ng/mL) were set to 0 for Phoenix PK calculations.

EXAMPLE 11: PREPARATION OF 5-CHLORO-2-(4-METHYLPHENYL)PYRIDINE (PROCESS ACCORDING TO SECTION A

(89) Under nitrogen 2,5-dichloro-pyridine (40 g, 270 mmol), 4-methylphenylboronic acid (39 g, 289 mmol) and bistriphenylphosphin-palladium(II) dichloride (1.14 g; 1.6 mmol) were suspended in water (258 g)/THF (117 g) for approx. 30 min at 35-55° C. A solution of tripotassium phosphate (143.4 g, 676 mmol) in water (143 g) was added at 35-55° C. during approx. 60-120 min and 55° C. was maintained for another approx. 30-45 min. More tripotassium phosphate (22.9 g, 108 mmol) in water (22.9 g) was added over a period of approx. 30 min and the temperature was raised to 55-60° C. to complete the reaction within another approx. 2 h.

(90) For extractive palladium removal a solution of cysteine (ca.16 g) in water (115 g) was added to the reaction mixture at 60-55° C. After approx. 1 h at 55° C. the biphasic reaction mixture was clarified by filtration over a pad of cellflock filter aid (2-5 g) and a THF/water mixture (110 g/75 g) was used for rinsing. The layers of the combined filtrates were separated at 25° C. and the salt containing water layer was extracted with THF (1×57 g). The combined THF layers were diluted with ethanol 94% (195 g) and concentrated by distillation under reduced pressure (300-200 mbar) at a jacket temperature of 45° C. in order to remove the bulk of THF (175-250 g). To the remaining product solution further ethanol (97 g) was added and at 45-55° C. water (565 g) was gradually added over a period of approx. 60 min to induce and maintain crystallization. After 30 min the temperature was lowered to approx. 20° C. in approx. 90-120 min and after another hour at that temperature the solids were collected by filtration, washed with ethanol/water 1:2 and dried under reduced pressure to yield 5-chloro-2-(4-methylphenyl)pyridine (52.5 g; 95% of theory; purity>95%; Pd<25 ppm).

EXAMPLE 12: PREPARATION OF (R)-3-(6-(4-METHYLPHENYL)-PYRIDIN-3-YLOXY)-1-AZA BICYCLO[2.2.2]OCTANE IN FREE FORM AND FUMARATE SALT FORM (PROCESS ACCORDING TO SECTION B)

Example 12.1: Formation of Free Form

(91) Under nitrogen, to 3R-quinuclidinol (43.8 g, 0.34 mol) in DMSO (792 g) an approx. 20% THF solution of potassium tert-butoxide (210 g, 0.375 mol) was added and at approx. 40-45° C. under reduced pressure the THF solvent was distilled off. The temperature of the reaction mixture was raised to 90° C. and the solid 5-chloro-2-(4-methylphenyl)pyridine (61.2 g, 0.30 mol) was gradually added in at least 4 portions. The temperature was raised further to approx. 100-105° C. and after at least another 3 hours at this temperature the reaction to (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane was complete.

(92) Water (150 g) was added to the reaction mixture at 60-25° C. and the temperature was gradually lowered to approx. 20° C. in approx. 60 min and additional water (210 g) was added. After at least another 2 further hours at this temperature the fine solids were collected by filtration, washed successively with DMSO/water (approx. 322 g; 2:1 mixture), water (500 g) and water/ethanol (approx. 500 g; 9:1 mixture) and dried at 60° C. under reduced pressure to yield (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane (56.3 g, 63% of theory).

Example 12.2: Formation of Fumarate Salt Form

(93) To a clear solution of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane (39.6 g; 0.135 mol) and fumaric acid (16.4 g, 0.141 mol) in ethanol (330 g)/water (21 g) at 65° C. tert.-butylmethylether (142.5 g) was added and the reaction mixture was cooled to 23° C. in approx. 60 min. Further tert.-butylmethylether (170.6 g) was added. After at least another 2 hours the solids were collected by filtration, washed with ethanol/tert.butylmethylether (153 g; 1.1 mixture) and dried at 55-60° C. under reduced pressure to yield (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane hydrogenfumarate (43.8 g, 79% of theory).

EXAMPLE 13: PREPARATION OF (R)-3-(6-(4-METHYLPHENYL)-PYRIDIN-3-YLOXY)-1-AZA BICYCLO[2.2.2]OCTANE IN FREE FORM AND FUMARATE SALT FORM (PROCESS ACCORDING TO SECTION B1)

Example 13.1: Formation of Free Form

(94) Under nitrogen to 3R-quinuclidinol (41.4 g, 0.325 mol) in DMSO (320 g) a solution of 5-chloro-2-(4-methylphenyl)pyridine (51 g, 0.250 mol) in toluene (201 g) was added. The temperature was raised gradually to approx. 100-105° C. while residual water, if any, was removed by refluxing under reduced pressure at a water trap for ca. 45 min. Over a period of approx. 90 min an approx. 20% THF solution of potassium tert-butoxide (158.8 g, 0.283 mol) was continuously added while gradually the THF solvent distilled off. After another 2-5 hours at approx. 100-105° C. the reaction to (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane was complete.

(95) Water (293 g) was added to the reaction mixture at 60-25° C. The layers were separated and the toluene layer was washed with water (2×42 g). The toluene solution was dried at ca. 60° C. by refluxing under reduced pressure at a water trap for ca. 45-60 min.

Example 13.2: Formation of Fumarate Salt Form

(96) To the toluene solution of Example 13.1, at ca. 50-55° C., a slurry of fumaric acid (26.1 g, 0.9 eq) in EtOH 94% (22 g) and toluene (97 g) was gradually added. Further toluene (97 g) was added for rinsing and after another ca. 30-60 min at 55° C. the temperature was gradually lowered to approx. 20° C. in approx. 120-180 min. After at least another 1 hour the solids were collected by filtration, washed with water saturated toluene (2×104 g) and dried at 60° C. under reduced pressure to yield (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane hydrogenfumarate (84.8 g; 82% of theory, based on amount of 5-chloro-2-(4-methylphenyl)pyridine used in Example 13.1).

BRIEF DESCRIPTION OF THE DRAWINGS

(97) FIG. 1 shows the XRPD pattern for the mono-fumarate salt of (R)-3-(6-(4-methylphenyl)pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in crystalline form

(98) FIG. 2 shows the XRPD pattern for the mono-maleate salt of (R)-3-(6-(4-methylphenyl)pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in crystalline form

(99) FIG. 3 shows the XRPD pattern for the mono-hydrochloride salt of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in crystalline form

(100) FIGS. 4A, 4B and 4C show the XRPD patterns for Form A, B and C of the phosphate salt of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in crystalline form

(101) FIGS. 5A and 5B show the XRPD patterns for Form A and B of the mono-succinate salt of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in crystalline form

(102) FIG. 6 shows the XRPD pattern for the mono-malonate salt of (R)-3-(6-(4-methylphenyl)pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in crystalline form

(103) FIGS. 7A and 7B show the XRPD patterns for Form A and B of the free base of (R)-3-(6-(4-methylphenyl)-pyridin-3-yloxy)-1-aza-bicyclo[2.2.2]octane in crystalline form