Piperidine inhibitors of Janus kinase 3

Abstract

The present invention relates to new piperidine inhibitors of Janus kinase 3 activity, pharmaceutical compositions thereof, and methods of use thereof. ##STR00001##

Claims

1. A compound that is ##STR00170## or a pharmaceutically acceptable salt thereof; wherein each position represented as D has deuterium enrichment of no less than about 10%.

2. The compound of claim 1, wherein each position represented as D has deuterium enrichment of no less than about 50%.

3. The compound of claim 1, wherein each position represented as D has deuterium enrichment of no less than about 90%.

4. The compound of claim 1, wherein each position represented as D has deuterium enrichment of no less than about 98%.

5. A pharmaceutical composition comprising a compound of claim 1, or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier.

6. The compound of claim 1 that is ##STR00171## or a pharmaceutically acceptable salt thereof.

7. The compound of claim 6, wherein each position represented as D has deuterium enrichment of no less than about 50%.

8. The compound of claim 6, wherein each position represented as D has deuterium enrichment of no less than about 90%.

9. The compound of claim 6, wherein each position represented as D has deuterium enrichment of no less than about 98%.

10. A pharmaceutical composition comprising a compound of claim 6, or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier.

11. The compound of claim 1 that is ##STR00172## or a pharmaceutically acceptable salt thereof.

12. The compound of claim 11, wherein each position represented as D has deuterium enrichment of no less than about 50%.

13. The compound of claim 11, wherein each position represented as D has deuterium enrichment of no less than about 90%.

14. The compound of claim 11, wherein each position represented as D has deuterium enrichment of no less than about 98%.

15. A pharmaceutical composition comprising a compound of claim 11, or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier.

16. The compound of claim 1 that is ##STR00173## or a pharmaceutically acceptable salt thereof.

17. The compound of claim 16, wherein each position represented as D has deuterium enrichment of no less than about 50%.

18. The compound of claim 16, wherein each position represented as D has deuterium enrichment of no less than about 90%.

19. The compound of claim 16, wherein each position represented as D has deuterium enrichment of no less than about 98%.

20. A pharmaceutical composition comprising a compound of claim 16, or a pharmaceutically acceptable salt thereof, together with a pharmaceutically acceptable carrier.

21. A pharmaceutical composition comprising a compound that is ##STR00174## or a combination thereof; or a pharmaceutically acceptable salt thereof; together with a pharmaceutically acceptable carrier; wherein each position represented as D has deuterium enrichment of no less than about 10%.

22. A method of treating a disorder in a patient wherein the disorder is renal transplant rejection, rheumatoid arthritis, psoriasis, inflammatory bowel disease, dry eye syndrome, asthma, or transplant rejection comprising administering to the patient therapeutically effective amount of a compound of claim 1.

23. The method of claim 22, wherein the disorder is renal transplant rejection.

24. The method of claim 22, wherein the disorder is rheumatoid arthritis.

25. The method of claim 22, wherein the disorder is psoriasis.

26. The method of claim 22, wherein the disorder is inflammatory bowel disease.

27. The method of claim 22, wherein the disorder is dry eye syndrome.

28. The method of claim 22, wherein the disorder is asthma.

29. The method of claim 22, wherein the disorder is transplant rejection.

Description

EXAMPLE 1

3-((3R,4R)-4-Methyl-3-(methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile mono citrate salt (CP-690550 citrate salt)

(1) ##STR00028##
Step 1

(2) ##STR00029##

4-Chloro-7-tosyl-7H-pyrrolo[2,3-d]pyrimidine

(3) At about 0 C., sodium hydroxide (2 mol/L in water, 8 mL, 1.20 equiv.) was added to a solution of 4-methylbenzene-1-sulfonyl chloride (2.7 g, 13.9 mmol, 1.10 equiv.) and 4-chloro-7H-pyrrolo[2,3-d]pyrimidine (2 g, 12.8 mmol, 1.00 equiv.) in acetone (20 mL). The resulting solution was stirred at about 20 C. for about 6 hours. The solids were collected by filtration and washed with acetone/water to give the title product as a white solid (4.0 g; yield=97%). .sup.1H NMR (300 MHz, CDCl.sub.3) : 8.78 (s, 1H), 8.11 (d, J=8.4 Hz, 2H), 7.80 (d, J=4.2 Hz, 1H), 7.34 (d, J=8.4 Hz, 2H), 7.73 (d, J=4.2 Hz, 1H), 2.42 (s, 3H). LC-MS: m/z=308/310 (M+H).sup.+.

(4) Step 2

(5) ##STR00030##

Methyl 4-methylpyridin-3-ylcarbamate

(6) At about 0 C., potassium tert-butoxide (47 g, 420 mmol, 3.00 equiv.) was added in several batches to a solution of 4-methylpyridin-3-amine (15 g, 139 mmol, 1.00 equiv.) in tetrahydrofuran (400 mL). After stirring the solution for about 30 minutes, dimethyl carbonate (18.8 g, 209 mmol, 1.50 equiv.) was then added. The solution was stirred at ambient temperature for about 16 hours and then water (100 mL) was added. Following standard extractive workup with ethyl acetate (3200 mL), the crude product was purified by re-crystallization from ethyl acetate/petroleum ether (1:1) to give the title product as a pale yellow solid (17 g; yield=74%). LC-MS: m/z=167 (M+H).sup.+.

(7) Step 3

(8) ##STR00031##

1-Benzyl-3-methoxycarbonylamino-4-methyl-pyridinum bromide

(9) 1-(Bromomethyl)benzene (19 g, 111 mmol, 1.10 equiv.) was added to a solution of methyl 4-methylpyridin-3-ylcarbamate (17 g, 102 mmol, 1.00 equiv.) in toluene (500 mL). The solution was stirred at about 110 C. for about 16 hours. After cooling to ambient temperature, the solids were collected by filtration and washed with toluene to afford the title product as a light brown solid (35 g; yield=97%).

(10) Step 4

(11) ##STR00032##

Methyl 1-benzyl-4-methyl-1,2,5,6-tetrahydropyridin-3-ylcarbamate

(12) Sodium borohydride (4.4 g, 116 mmol, 1.20 equiv.) was added in several batches to a solution of 1-benzyl-3-methoxycarbonylamino-4-methyl-pyridinum bromide (35 g, 104 mmol, 1.00 equiv.) in methanol (300 mL). The resulting solution was stirred at ambient temperature for about 16 hours, and then water (200 mL) was added. After concentrating the mixture in vacuo, standard extractive workup with ether (3200 mL) gave a crude residue that was then purified by silica gel column chromatography (dichloromethane/methanol (20:1)) to afford the title product as a yellow solid (18 g; yield=66%). LC-MS: m/z=261 (M+H).sup.+.

(13) Step 5

(14) ##STR00033##

Methyl 1-benzyl-4-methylpiperidin-3-yl-carbamate

(15) Platinum oxide (1.0 g, 4.41 mmol, 0.11 equiv.) was added to a solution of methyl 1-benzyl-4-methyl-1,2,5,6-tetrahydropyridin-3-ylcarbamate (10 g, 38.46 mmol, 1.00 equiv.) in methanol (200 mL). After introducing hydrogen gas, the mixture was stirred at about 60 C. for about 16 hours and then was filtered. The resulting filtrate was concentrated to give a crude residue that was then purified by silica gel column chromatography (ethyl acetate/petroleum (1:2)) to afford the title product a yellow solid (7 g; yield=66%). LC-MS: m/z=263 (M+H).sup.+.

(16) Step 6

(17) ##STR00034##

(1-Benzyl-4-methyl-piperidin-3-yl)-methyl-amine

(18) At about 0 C., lithium aluminum hydride (3.6 g, 92.8 mmol, 5.00 equiv.) was added in several batches to a solution of methyl 1-benzyl-4-methylpiperidin-3-yl-carbamate (5.0 g, 18.1 mmol, 1.00 equiv.) in tetrahydrofuran (100 mL). The resulting solution was heated at reflux for about 16 hours, and then water (10 mL) was added. The mixture was filtered, and the resulting filtrate was concentrated in vacuo to give a crude residue that was then purified by silica gel column chromatography (dichloromethane/methanol (20:1)) to afford the title product as a yellow oil (3.0 g; yield=72%). .sup.1H NMR (300 MHz, CDCl.sub.3) : 7.20-7.38 (m, 5H), 3.58 (d, J=13.2 Hz, 1H), 3.48 (d, J=13.2 Hz, 1H), 2.60-2.82 (m, 2H), 2.46 (br s, 1H), 2.34 (s, 3H), 2.02-2.22 (m, 2H), 2.64-2.84 (m, 2H), 1.45-1.58 (m, 2H), 0.97 (d, J=6.9 Hz, 3H). LC-MS: m/z=219 (M+H).sup.+.

(19) Step 7

(20) ##STR00035##

N-(1-Benzyl-4-methylpiperidin-3-yl)-N-methyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

(21) 4-Chloro-7-tosyl-7H-pyrrolo[2,3-d]pyrimidine (2 g, 6.37 mmol, 2.00 equiv.) and potassium carbonate (2.7 g, 19.4 mmol, 6.00 equiv.) were added to a solution of (1-benzyl-4-methyl-piperidin-3-yl)-methyl-amine (700 mg, 2.89 mmol, 1.00 equiv.) in water (30 mL). The solution was stirred at about 100 C. for about 16 hours, and then was cooled to ambient temperature. Following standard extractive workup with ethyl acetate (3100 mL), the crude residue was purified by silica gel column chromatography (ethyl acetate/petroleum (1:1)) to give the title product as a light yellow solid (1.5 g; yield=96%). .sup.1H NMR (300 MHz, CDCl.sub.3) : 8.36 (s, 1H), 8.08 (d, J=8.4 Hz, 2H), 7.45 (d, J=4.2 Hz, 1H), 7.20-7.42 (m, 7H), 6.75 (d, J=4.2 Hz, 1H), 5.05-5.20 (m, 1H), 3.40-3.65 (m, 5H), 2.70-2.92 (m, 2H), 2.50-2.70 (m, 1H), 2.42 (s, 3H), 2.23-2.42 (m, 1H), 2.10-2.23 (m, 1H), 1.55-1.75 (m, 2H), 0.92 (d, J=6.9 Hz, 3H). LC-MS: m/z=490 (M+H).sup.+.

(22) Step 8

(23) ##STR00036##

N-(1-Benzyl-4-methylpiperidin-3-yl)-N-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

(24) A mixture of 50% sodium hydroxide (10 mL) and N-(1-benzyl-4-methylpiperidin-3-yl)-N-methyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (400 mg, 0.80 mmol, 1.00 equiv.) was stirred at about 60 C. for about 16 hours, and then was cooled to ambient temperature. Following standard extractive workup with ethyl acetate (410 mL), the crude residue was then purified by silica gel column chromatography (dichloromethane/methanol (10:1)) to give the title product as a yellow solid (0.25 g; yield=88%). .sup.1H NMR (300 MHz, CDCl.sub.3) : 11.35 (br s, 1H), 8.30 (s, 1H), 7.20-7.40 (m, 5H), 7.06 (d, J=3.6 Hz, 1H), 6.60 (d, J=3.6 Hz, 1H), 5.20-5.30 (m, 1H), 3.66 (s, 3H), 3.48-3.65 (m, 2H), 2.85-2.98 (m, 1H), 2.60-2.85 (m, 2H), 2.30-2.45 (m, 1H), 2.12-2.30 (m, 1H), 1.60-1.92 (m, 2H), 0.98 (d, J=6.0 Hz, 3H). LC-MS: m/z=336 (M+H).sup.+.

(25) Step 9

(26) ##STR00037##

N-((3R,4R)-1-Benzyl-4-methylpiperidin-3-yl)-N-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

(27) The enantiomer N-((3R,4R)-1-benzyl-4-methylpiperidin-3-yl)-N-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (4.5 g) was isolated by chiral resolution using chiral-Prep-HPLC with the following conditions: Column: Chiralpak IA, 0.4625 cm; mobile phase: hexane (in 0.1% triethylamine): isopropanol (90:10); Detector: UV 254 nm. Retention time of desired enantiomer: 11.72 minutes, undesired enantiomer retention time: 7.88 minutes. ee %>99.8%. The title product was isolated a yellow solid (1.8 g; yield=40%). .sup.1H NMR (300 MHz, CDCl.sub.3) : 11.35 (br s, 1H), 8.30 (s, 1H), 7.20-7.40 (m, 5H), 7.06 (d, J=3.6 Hz, 1H), 6.60 (d, J=3.6 Hz, 1H), 5.20-5.30 (m, 1H), 3.66 (s, 3H), 3.48-3.65 (m, 2H), 2.85-2.98 (m, 1H), 2.60-2.85 (m, 2H), 2.30-2.45 (m, 1H), 2.12-2.30 (m, 1H), 1.60-1.92 (m, 2H), 0.98 (d, J=6.0 Hz, 3H). LC-MS: m/z=336 (M+H).sup.+.

(28) Step 10

(29) ##STR00038##

N-Methyl-N-((3R,4R)-4-methylpiperidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine

(30) Palladium hydroxide on carbon (50 mg), and acetic acid (44 mg, 0.72 mmol, 1.00 equiv.) were added to a solution of N-((3R,4R)-1-benzyl-4-methylpiperidin-3-yl)-N-methyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (250 mg, 0.67 mmol, 1.00 equiv.) in isopropanol/water (10 mL/2 mL). After hydrogen gas was introduced, the resulting mixture was stirred at about 50 C. for about 16 hours. After filtering the mixture, the pH value of the filtrate was adjusted to 8 by adding sodium hydroxide. Standard extractive workup with dichloromethane (320 mL) afforded the title product as an off-white solid (140 mg; yield=81%) .sup.1H NMR (300 MHz, CDCl.sub.3) : 10.60 (br s, 1H), 8.35 (s, 1H), 7.07 (d, J=3.6 Hz, 1H), 6.60 (d, J=3.6 Hz, 1H), 4.88-4.98 (m, 1H), 3.45 (s, 3H), 3.25-3.37 (m, 1H), 2.80-3.10 (m, 3H), 2.45-2.58 (m, 1H), 1.82-2.00 (m, 1H), 1.60-1.80 (m, 2H), 1.11 (d, J=7.2 Hz, 3H). LC-MS: m/z=246 (M+H).sup.+.

(31) Step 11

(32) ##STR00039##

3-((3R,4R)-4-Methyl-3-(methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile (CP-690550)

(33) Ethyl 2-cyanoacetate (140 mg, 1.23 mmol, 6.00 equiv.) and triethylamine (40 mg, 0.39 mmol, 2.00 equiv.) were added to a solution of N-methyl-N-((3R,4R)-4-methylpiperidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine 12 (50 mg, 0.19 mmol, 1.00 equiv.) in toluene (10 mL). The resulting solution was stirred at about 110 C. for about 16 hours and then was concentrated in vacuo. The resulting residue was purified by silica gel column chromatography (ethyl acetate/methanol (50:1)) to give the title product as a light yellow solid (33 mg; yield=52%). .sup.1H NMR (300 MHz, CD.sub.3OD) : 8.10 (s, 1H), 7.10 (d, J=4.0 Hz, 1H), 6.65 (d, J=4.0 Hz, 1H), 5.00-5.10 (m, 1H), 3.80-4.00 (m, 2H), 3.55-3.75 (m, 1H), 3.40-3.55 (m, 1H), 3.30-3.40 (m, 5H), 2.40-2.55 (m, 1H), 1.82-2.00 (m, 1H), 1.60-1.80 (m, 1H), 1.05-1.20 (m, 3H). LC-MS: m/z=313 (M+H).sup.+.

(34) Step 12

(35) ##STR00040##

3-((3R,4R)-4-Methyl-3-(methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile mono citrate salt (CP-690550 citrate salt)

(36) Citric acid (20 mg, 0.10 mmol, 1.00 equiv.) was added to a solution of 3-(3R,4R)-4-methyl-3-(methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile (33 mg, 0.10 mmol, 1.00 equiv.) in water/methanol (5/0.5 mL). The resulting solution was stirred at about 40 C. for about 10 minutes, and then was cooled to ambient temperature. The solvent was then removed by using a cryofreeze-dryer to give the title compound as an off-white solid (40 mg; yield=76%). .sup.1H NMR (300 MHz, CD.sub.3OD) : 8.15 (s, 1H), 7.15 (d, J=3.6 Hz, 1H), 6.70 (d, J=3.6 Hz, 1H), 4.95-5.15 (m, 1H), 3.85-4.08 (m, 4H), 3.58-3.80 (m, 1H), 3.40-3.60 (m, 4H), 2.92 (Ab.sub.q, J=15.6 Hz, 2H), 2.80 (Ab.sub.q, J=15.6 Hz, 2H), 2.40-2.60 (m, 1H), 1.85-2.05 (m, 1H), 1.68-1.85 (m, 1H), 1.05-1.20 (m, 3H). LC-MS: m/z=313 (MH-C.sub.6H.sub.8O.sub.7).sup.+.

EXAMPLE 2

3-((3R,4R)-4-Methyl-3-(d3-methyl(2-d1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile mono citrate salt (CP-690550-d4 citrate salt)

(37) ##STR00041##
Step 1

(38) ##STR00042##

4-Chloro-7-tosyl-7H-pyrrolo[2,3-d]pyrimidine

(39) 4-Methylbenzene-1-sulfonyl chloride (3.7 g, 19.32 mmol, 1.20 equiv.) was added to a solution of 2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidine 1 (3 g, 16.1 mmol, 1.00 equiv.) in acetone (20 mL). At about 0 C., an aqueous sodium hydroxide solution (2 mol/L, 12 mL) was added dropwise to the solution. The solution was then stirred at ambient temperature for about 3 hours. The solids were collected by filtration and washed with acetone/water to give the title product as a white solid (5.2 g; yield=95%). LC-MS: m/z=342 (M+H).sup.+.

(40) Step 2

(41) ##STR00043##

(1-Benzyl-4-methyl-piperidin-3-yl)-d3-methyl-amine

(42) The procedure of Example 1, Step 6 was followed but substituting lithium aluminum deuteride for lithium aluminum hydride. The title product was isolated as a yellow oil (3.0 g; yield=72%). LC-MS: m/z=222 (M+H).sup.+.

(43) Step 3

(44) ##STR00044##

N-(1-Benzyl-4-methylpiperidin-3-yl)-2-chloro-N-d3-methyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

(45) A mixture of (1-benzyl-4-methyl-piperidin-3-yl)-d.sub.3-methyl-amine (700 mg, 2.89 mmol, 1.00 equiv.), 2,4-dichloro-7H-pyrrolo [2, 3-d]-pyrimidine (2 g, 5.78 mmol, 2.00 equiv.) and potassium carbonate (2.7 g, 19.4 mmol, 6.00 equiv) in tetrahydrofuran/water (1:1) (60 mL) was heated at about 60 C. for about 16 hours, and then the solvent was removed in vacuo. Following standard extractive workup with ethyl acetate (3200 mL), the crude residue was purified by column chromatography to give the title product as a light yellow solid (1.5 g; yield=96%). LC-MS: m/z=527 (M+H).sup.+.

(46) Step 4

(47) ##STR00045##

tert-Butyl 4-methyl-3-(d3-methyl(2-d1-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate

(48) Under an atmosphere of deuterium gas, a solution of N-(1-benzyl-4-methylpiperidin-3-yl)-2-chloro-N-d.sub.3-methyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine (400 mg, 0.80 mmol, 1.00 equiv.), di-tert-butyl dicarbonate (348 mg, 1.6 mmol) and palladium hydroxide on carbon (1.00 equiv.; pre-treated with deuterium oxide for three cycles) in d.sub.4-methanol/deuterium oxide (1:3) (30 mL) was heated at about 70 C. for about 16 hours. Following standard extractive workup with ethyl acetate, the crude residue was purified by silica gel column chromatography to give the title product as a solid (300 mg; yield=78.5%). LC-MS: m/z=504 (M+H).sup.+.

(49) Step 5

(50) ##STR00046##

4-Methyl-3-[d3-methyl-(2-d1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidine-1-carboxylic acid tert-butyl ester

(51) A solution of tert-butyl 4-methyl-3-(d.sub.3-methyl(2-d.sub.1-7-tosyl-7H-pyrrolo [2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate (300 mg) in 30% d.sub.1-sodium hydroxide (60 mL) was heated at about 100 C. for about 2 hours. Following standard extractive workup with ethyl acetate (3200 mL), the crude residue was purified by silica gel column chromatography to afford the title product as a foamy solid (190 mg; yield=90%). LC-MS: m/z=350 (M+H).sup.+.

(52) Step 6

(53) ##STR00047##

3-((3R,4R)-4-Methyl-3-[d3-methyl-(2-d1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidine)-1-carboxylic acid tert-butyl ester

(54) The enantiomer 3-((3R,4R)-4-methyl-3-[d.sub.3-methyl-(2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidine)-1-carboxylic acid tert-butyl ester was isolated from 4-methyl-3-[d.sub.3-methyl-(2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidine-1-carboxylic acid tert-butyl ester (4.5 g) by chiral resolution using chiral-Prep-HPLC with the following conditions: column, Chiralpak IA, 0.4615 cm; mobile phase: (hexane:isopropyl alcohol (90:10)); detector: UV 254 nm. Retention time of desired enantiomer: 7.19 minutes, undesired enantiomer retention time: 9.11 minutes. ee %>99.8%. The title product was isolated as a yellow solid (1.5 g; yield=35%). LC-MS: m/z=527 (M+H).sup.+.

(55) Step 7

(56) ##STR00048##

N-d3-Methyl-N-((3R,4R)-4-methylpiperidin-3-yl)-2-d1-7H-pyrrolo[2,3-d]pyrimidin-4-amine deuterochloride

(57) A solution of 3-((3R,4R)-4-methyl-3-[d.sub.3-methyl-(2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidine)-1-carboxylic acid tert-butyl ester (190 mg) in 5N deuterium chloride/dioxane (0.5 mL/3 mL) was stirred at 25 C. for about 16 hours. The solution was concentrated in vacuo, and the resulting residue was used in the next step without any further purification. LC-MS: m/z=250 (M+H).sup.+.

(58) Step 8

(59) ##STR00049##

3-((3R,4R)-4-Methyl-3-(d3-methyl(2-d1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)amino)piperidin-1-yl)-3-oxopropanenitrile (CP-690550)

(60) The procedure of Example 1, Step 11 was followed, but substituting N-d.sub.3-methyl-N-((3R,4R)-4-methylpiperidin-3-yl)-2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-amine deuterochloride for N-methyl-N-((3R,4R)-4-methylpiperidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine. The title product was isolated as a light yellow solid (33 mg; yield=52%). LC-MS: m/z=317 (M+H).sup.+.

(61) Step 9

(62) ##STR00050##

3-((3R,4R)-4-Methyl-3-(d3-methyl(2-d1-7H-pyrrolo[2,3-d]pyrimidin-4-1)amino)piperidin-1-yl)-3-oxopropanenitrile mono citrate salt (CP-690550-d4 citrate salt)

(63) The procedure of Example 1, Step 12 was followed, but substituting 3-((3R,4R)-4-methyl-3-(d.sub.3-methyl(2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile for 3-((3R,4R)-4-methyl-3-(methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile. The title product was isolated as a white solid (40 mg; yield=76%). .sup.1H NMR (300 MHz, CD.sub.3OD) : 7.36 (s, 1H), 6.89 (s, 1H), 4.95-5.15 (m, 1H), 3.85-4.08 (m, 4H), 3.48-3.75 (m, 2H), 2.94 (Ab.sub.q, J=15.9 Hz, 2H), 2.81 (Ab.sub.q, J=15.6 Hz, 2H), 2.48-2.61 (m, 1H), 1.89-2.05 (m, 1H), 1.69-1.88 (m, 1H), 1.14 (d, J=6.6 Hz, 3H). LC-MS: m/z=317 (MH-C.sub.6H.sub.8O.sub.7).sup.+.

EXAMPLE 3

3-((3R,4R)-4-d3-methyl-3-(d3-methyl (2-d1-7H-pyrr-olo [2, 3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile mono citrate salt (CP-690550-d7 citrate salt)

(64) ##STR00051##
Step 1

(65) ##STR00052##

Ethyl 3-oxopiperidine-4-carboxylate acetic salt

(66) Under an atmosphere of hydrogen, the mixture of ethyl 1-benzyl-3-oxopiperidine-4-carboxylate (20 g, 16.1 mmol, 1.00 equiv.), 10% palladium on carbon, acetic acid (10 mL), and methanol (100 mL) was heated at about 50 C. for about 4 hours. The mixture was filtered, the filtrate was evaporated to give the title product as an acetic salt (16 g; yield=85%). LC-MS: m/z=172 (M+H).sup.+.

(67) Step 2

(68) ##STR00053##

Methyl 4-methylpyridin-3-ylcarbamate

(69) A solution of di-tert-butyl dicarbonate (5.66 g, 26 mmol), potassium carbonate (12 g, 86.4 mmol) and water (100 mL) was added to a solution of ethyl 3-oxopiperidine-4-carboxylate acetic salt (15 g, 21.6 mmol) in tetrahydrofuran (400 mL). The resulting mixture was stirred at ambient temperature for about 2 hours. After removing the solvent in vacuo, standard extractive workup with ethyl acetate (3200 mL) afforded the title product as a pale white solid (14 g; yield=80%). LC-MS: m/z=172/272 (M+H).sup.+.

(70) Step 3

(71) ##STR00054##

1-tert-Butyl-4-ethyl 4-d3-methyl-3-oxopiperidine-1,4-dicarboxylate

(72) 70% Sodium hydride (3.54 g, 103 mmol) was added in several portions to a solution of 1-tert-butyl 4-ethyl 3-oxopiperidine-1, 4-dicarboxylate (14 g, 51.6 mmol, 1.00 equiv.) in tetrahydrofuran (300 mL). The resulting mixture was heated at about 50 C. for about 2 hours, and then was cooled to ambient temperature. After adding iodomethane (15 g, 103 mmol), the resulting suspension was stirred at ambient temperature for about 3 hours and then poured into ice. Standard extractive workup with ethyl acetate (3100 mL) gave a crude residue that was then purified by column chromatography to give the title product as a solid (7.4 g; yield=50%). LC-MS: m/z=289 (M+H).sup.+.

(73) Step 4

(74) ##STR00055##

4-d3-Methylpiperidin-3-one hydrochloride

(75) 37% Hydrogen chloride (30 mL) was added to 1-tert-butyl 4-ethyl 4-d.sub.3-methyl-3-oxopiperidine-1,4-dicarboxylate (7 g, 25.6 mmol). The resulting mixture was heated at reflux for about 3 hours, and then the solvent was removed by evaporation in vacuo. The resulting residue was used in the next step without further purification. LC-MS: m/z=117/125 (M+H).sup.+.

(76) Step 5

(77) ##STR00056##

1-Benzyl-4-d3-methylpiperidin-3-one

(78) (Bromomethyl)benzene (2.23 g, 10.5 mmol) was added dropwise to a solution of 4-d.sub.3-methylpiperidin-3-one hydrochloride (1.2 g, 10.3 mmol, 1.00 equiv.) and triethylamine (2.1 g, 20.6 mmol) in tetrahydrofuran (30 mL). The resulting mixture was stirred at ambient temperature for about 16 hours, and then solvent was evaporated in vacuo. Following standard extractive workup with ethyl acetate, the crude residue was purified by column chromatography to give the title product as a solid (1.7 g; yield=80%). .sup.1H NMR (300 MHz, CD.sub.3OD) : 7.21-7.39 (m, 5H), 3.5 (s, 2H), 3.23 (d, J=13.8 Hz, 1H), 2.94 (d, J=9.6 Hz, 1H), 2.79 (d, J=13.8 Hz, 1H), 2.45 (t, J=11.4 Hz, 1H), 2.29-2.39 (m, 1H), 1.98-2.01 (m, 1H), 1.59-1.71 (m, 341H). LC-MS: m/z=207/225 (M+H).sup.+.

(79) Step 6

(80) ##STR00057##

(1-Benzyl-4-d3-methyl-piperidin-3-yl)-d3-methyl-amine

(81) At about 0 C., sodium methoxide (3.2 g, 38.2 mmol) was added to a suspension of d.sub.3-methylamine hydrochloride (1.4 g, 19.4 mmol), 1-benzyl-4-d.sub.3-methylpiperidin-3-one (2 g 9.7 mmol) and tetrahydrofuran (60 mL). The mixture was stirred at ambient temperature for about 16 hours, and then sodium triacetoxy borohydride (8.5 g, 40 mmol) was added. The mixture was stirred at ambient temperature for about 5 hours, and then 5% sodium hydroxide (50 mL) was added. Following standard extractive workup with ethyl acetate, the crude residue was purified by silica gel column chromatography (dichloromethane/methanol) to afford the title product (2.2 g; yield=50%). LC-MS: m/z=225 (M+H).sup.+.

(82) Step 7

(83) ##STR00058##

N-(1-Benzyl-4-d3-methylpiperidin-3-yl)-2-chloro-N-d3-methyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

(84) The procedure of Example 2, Step 3 was followed but substituting (1-benzyl-4-d.sub.3-methyl-piperidin-3-yl)-d.sub.3-methyl-amine for (1-benzyl-4-methyl-piperidin-3-yl)-d.sub.3-methyl-amine. The title product was isolated a light yellow solid (1.4 g; yield=90%). LC-MS: m/z=530 (M+H).sup.+.

(85) Step 8

(86) ##STR00059##

tert-Butyl 4-d3-methyl-3-(d3-methyl(2-d1-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate

(87) The procedure of Example 2, Step 4 was followed but substituting N-(1-benzyl-4-d.sub.3-methylpiperidin-3-yl)-2-chloro-N-d.sub.3-methyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine for N-(1-benzyl-4-methylpiperidin-3-yl)-2-chloro-N-d.sub.3-methyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine. The title product was isolated as a solid (270 mg, yield=70%). LC-MS: m/z=507 (M+H).sup.+.

(88) Step 9

(89) ##STR00060##

tert-Butyl 4-((1-(tert-butoxycarbonyl)-4-d3-methylpiperidin-3-yl)-d3-methyl)amino)-2-d1-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate

(90) A mixture of tert-butyl 4-d.sub.3-methyl-3-(d.sub.3-methyl(2-d.sub.1-7-tosyl-7H-pyrrolo [2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate (200 mg, 0.4 mmol) and 30% d.sub.1-sodium hydroxide (60 mL) was heated at about 100 C. for about 16 hours. After cooling the mixture to ambient temperature, di-tert-butyl dicarbonate (170 mg, 0.8 mmol) and tetrahydrofuran (20 ml) were added. The mixture was stirred at ambient temperature for about 16 hours, and then the solvent was removed in vacuo. Following standard extractive workup with ethyl acetate, the resulting crude residue was purified by silica gel column chromatography (ethyl acetate/petroleum (1:5)) to give the title product as a white solid. LC-MS: m/z=453 (M+H).sup.+.

(91) Step 10

(92) ##STR00061##

(3R,4R)-tert-Butyl 4-((1-(tert-butoxycarbonyl)-4-d3-methylpiperidin-3-yl)(d3-methyl)amino)-2-d1-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate

(93) The enantiomer (3R,4R)-tert-butyl 4-((1-(tert-butoxycarbonyl)-4-d.sub.3-methylpiperidin-3-yl)(d.sub.3-methyl)amino)-2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate was isolated from tert-butyl 4-((1-(tert-butoxycarbonyl)-4-d.sub.3-methylpiperidin-3-yl)(d.sub.3-methyl)amino)-2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidine-7-carboxy-late (300 mg) by chiral resolution using chiral-Prep-HPLC with the following conditions: column: Chiralpak IA (Waters 2767-1), 0.4625 cm; mobile phase: hexane/isopropyl alcohol (90:10); detector: UV 254 nm. Retention time of desired enantiomer: 6.08 minutes, undesired enantiomer retention time: 10.16 minutes. ee %>99.8%. The title product was isolated as a white solid (0.1 g; yield=35%). LC-MS: m/z=353 (M+H).sup.+.

(94) Step 11

(95) ##STR00062##

(96) N-d.sub.3-Methyl-N-((3R,4R)-4-d.sub.3-methylpiperidin-3-yl)-2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-amine deuterochloride: The procedure of Example 2, Step 7 was followed, but substituting (3R,4R)-tert-butyl 4-((1-(tert-butoxycarbonyl)-4-d.sub.3-methylpiperidin-3-yl)(d.sub.3-methyl)amino)-2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidine-7-carboxylate for 3-((3R,4R)-4-methyl-3-[d.sub.3-methyl-(2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidine)-1-carboxylic acid tert-butyl ester. The title product was isolated as a crude residue and was used in the next step without any further purification. LC-MS: m/z=253 (M+H).sup.+.

(97) Step 12

(98) ##STR00063##

3-((3R,4R)-4-d3-Methyl-3-(d3-methyl(1-d1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)amino)piperidin-1-yl)-3-oxopropanenitrile (CP-690550-d7)

(99) The procedure of Example 1, Step 11 was followed but substituting N-d.sub.3-methyl-N-((3R,4R)-4-d.sub.3-methylpiperidin-3-yl)-2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-amine deuterochloride for N-methyl-N-((3R,4R)-4-methylpiperidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine. The title product was isolated as a light yellow solid (40 mg; yield=56%). LC-MS: m/z=320 (M+H).sup.+.

(100) Step 13

(101) ##STR00064##

3-((3R,4R)-4-d3-Methyl-3-(d3-methyl(2-d1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile mono citrate salt (CP-690550-d7 citrate salt)

(102) The procedure of Example 1, Step 12 was followed but substituting 3-((3R,4R)-4-d.sub.3-methyl-3-(d.sub.3-methyl(2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile for 3-((3R,4R)-4-methyl-3-(methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile. The title product was isolated as an off-white solid (23 mg; yield=41%). .sup.1H NMR (300 MHz, CD.sub.3OD) : 7.36 (s, 1H), 6.89 (s, 1H), 4.95-5.15 (m, 1H), 3.85-4.08 (m, 4H), 3.48-3.75 (m, 2H), 2.94 (Ab.sub.q, J=15.6 Hz, 2H), 2.81 (Ab.sub.q, J=15.9 Hz, 2H), 2.48-2.61 (m, 1H), 1.89-2.05 (m, 1H), 1.69-1.88 (m, 1H). LC-MS: m/z=320 (MH-C.sub.6H.sub.8O.sub.7).sup.+.

EXAMPLE 4

3-((3R,4R)-4-d3-methyl-3-(d3-methyl (2-d1-7H-pyrrolo [2, 3-d]pyrimidin-4-yl)amino)-2, 2, 3, 4-d1-piperidin-1-yl)-3-oxo-propanenitrile mono citrate salt (CP-690550-d11 citrate salt)

(103) ##STR00065##
Step 1

(104) ##STR00066##

1-Benzyl-4-d3-methyl-(2,2,4-d3)-piperidin-3-one

(105) A mixture of 1-benzyl-4-d.sub.3-methylpiperidin-3-one 6 (2.5 g, 12.1 mmol) in 2N deuterium chloride in deuterium oxide (60 mL) was heated at about 80 C. for about 16 hours. After cooling the mixture to ambient temperature, 2N d.sub.1-sodium hydroxide in deuterium oxide (80 mL) was added. Standard extractive workup with ethyl acetate, gave a crude residue which was used in the next step without further purification. LC-MS: m/z=210/228 (M+H).sup.+.

(106) Step 2

(107) ##STR00067##

(1-Benzyl-4-d3-methyl-2,2,3,4-d4-piperidin-3-yl)-d3-methyl-amine

(108) The procedure of Example 3, Step 6 was followed but substituting 1-benzyl-4-d.sub.3-methyl-(2,2,4-d.sub.3)-piperidin-3-one for 1-benzyl-4-d.sub.3-methyl-piperidin-3-one. The title product was isolated as a solid (3.9 g; yield=90%). LC-MS: m/z=229 (M+H).sup.+.

(109) Step 3

(110) ##STR00068##

N-(1-Benzyl-4-d3-methyl-2,2,3,4-d4-piperidin-3-yl)-2-chloro-N-d3-methyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

(111) The procedure of Example 2, Step 3 was followed, but substituting (1-benzyl-4-d.sub.3-methyl-2,2,3,4-d.sub.4-piperidin-3-yl)-d.sub.3-methyl-amine for (1-Benzyl-4-methyl-piperidin-3-yl)-d.sub.3-methyl-amine. The title product was isolated as a light yellow solid (1.4 g; yield=90%). LC-MS: m/z=534 (M+H).sup.+.

(112) Step 4

(113) ##STR00069##

tert-Butyl 4-d3-methyl-3-(d3-methyl(2-d1-7-tosyl-7H-pyrrolo [2,3-d]pyrimidin-4-yl)amino)-2,2, 3,4-d4-peridine-1-carboxylate

(114) The procedure of Example 2, Step 4 was followed, but substituting N-(1-benzyl-4-d.sub.3-methyl-2,2,3,4-d.sub.4-piperidin-3-yl)-2-chloro-N-d.sub.3-methyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine for N-(1-benzyl-4-methylpiperidin-3-yl)-2-chloro-N-d.sub.3-methyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine. The title product was isolated as a solid (270 mg; yield=70%). LC-MS: m/z=411/511 (M+H).sup.+.

(115) Step 5

(116) ##STR00070##

(117) tert-Butyl 4-d.sub.3-methyl-3-(d.sub.3-methyl(2-d.sub.1-7-tosyl-7H-pyrrolo [2,3-d]pyrimidin-4-yl) amino)-2,2,3,4-d.sub.4-piperidine-1-carboxy-late: The procedure of Example 2, Step 5 was followed, but substituting tert-butyl 4-d.sub.3-methyl-3-(d.sub.3-methyl(2-d.sub.1-7-tosyl-7H-pyrrolo [2,3-d]pyrimidin-4-yl)amino)-2,2,3,4-d.sub.4-piperidine-1-carboxylate for tert-butyl 4-methyl-3-(d.sub.3-methyl(2-d.sub.1-7-tosyl-7H-pyrrolo [2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate. The title product was isolated as a foamy solid (130 mg; yield=90%). .sup.1H NMR (300 MHz, CD.sub.3OD) : 10.41-10.73 (brs, 1H), 7.07 (d, J=3.6 Hz, 1H), 6.57 (d, J=2.4 Hz, 1H), 3.38-3.71 (brs, 2H), 1.76-1.91 (m, 1H), 1.58-1.65 (m, 1H), 1.47 (s, 9H). LC-MS: m/z=257/357 (M+H).sup.+.

(118) Step 6

(119) ##STR00071##

(3R,4R)-tert-Butyl-4-d3-methyl-3-(d3-methyl (2-d1-7H-pyrrolo [2,3-d] pyrimidin-4-yl)amino)-2,2,3,4-d4-piperidine-1-carboxylate

(120) The enantiomer (3R,4R)-tert-butyl-4-d.sub.3-methyl-3-(d.sub.3-methyl (2-d.sub.1-7H-pyrrolo [2,3-d] pyrimidin-4-yl)amino)-2,2,3,4-d.sub.4-piperidine-1-carboxylate was isolated from tert-butyl 4-d.sub.3-methyl-3-(d.sub.3-methyl(2-d.sub.1-7-tosyl-7H-pyrrolo [2,3-d]pyrimidin-4-yl) amino)-2,2,3,4-d.sub.4-piperidine-1-carboxy-late by chiral resolution using chiral-prep-HPLC with the following conditions: column: Chiralpak IC225 cm (Waters 2767-1), Sum Chiral-P(IC)001IC00CJ-LD016; mobile phase: hexane/isopropyl alcohol (85:15); detector: UV 254 nm. Retention time of desired enantiomer: 12.01 minutes, undesired enantiomer retention time: 15.10 minutes. ee %>99.8%. The title product was isolated as a yellow solid (0.1 g; yield=35%). LC-MS: m/z=490 (M+H).sup.+.

(121) Step 7

(122) ##STR00072##

N-d3-Methyl-N-((3R,4R)-4-d3-methyl-2,2,3,4-d4-piperidin-3-yl)-2-d1-7H-pyrrolo[2,3-d]pyrimidin-4-amine deuterochloride

(123) The procedure of Example 2, Step 7 was followed, but substituting (3R,4R)-tert-butyl-4-d.sub.3-methyl-3-(d.sub.3-methyl (2-d.sub.1-7H-pyrrolo [2,3-d]pyrimidin-4-yl)amino)-2,2,3,4-d.sub.4-piperidine-1-carboxylate for 3-((3R,4R)-4-methyl-3-[d.sub.3-methyl-(2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidine)-1-carboxylic acid tert-butyl ester. The title product was isolated and used in the next step without further purification. LC-MS: m/z=257 (M+H).sup.+.

Step 8

(124) ##STR00073##

3-((3R,4R)-4-d3-Methyl-3-(d3-methyl(2-d1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2,2,3,4-d4-piperidin-1-yl)-3-oxopropane-nitrile (CP-690550-d11)

(125) The procedure of Example 1, Step 11 was followed, but substituting N-d.sub.3-methyl-N-((3R,4R)-4-d.sub.3-methyl-2,2,3,4-d.sub.4-piperidin-3-yl)-2-d.sub.1-7H-pyrrolo[2,3-d] pyrimidin-4-amine deuterochloride for N-methyl-N-((3R,4R)-4-methylpiperidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine. The title product was isolated as a light yellow solid (50 mg; yield=63%). LC-MS: m/z=324 (M+H).sup.+.

(126) Step 9

(127) ##STR00074##

3-((3R, 4R)-4-d3-Methyl-3-(d3-methyl (2-d1-7H-pyrrolo [2, 3-d]pyrimidin-4-yl)amino))-2,2,3,4-d4-piperidin-1-yl)-3-oxopropane-nitrile mono citrate salt CP-690550-d11 citrate salt

(128) The procedure of Example 1, Step 12 was followed, but substituting 3-((3R,4R)-4-d.sub.3-methyl-3-(d.sub.3-methyl(2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2,2,3,4-d.sub.4-piperidin-1-yl)-3-oxopropane-nitrile for 3-((3R,4R)-4-methyl-3-(methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile. The title product was isolated as an off-white solid (50 mg; yield=80%). .sup.1H NMR (300 MHz, CD.sub.3OD) : 7.36 (s, 1H), 6.89 (s, 1H), 3.91-4.08 (m, 2H), 3.48-3.75 (m, 2H), 2.94 (Ab.sub.q, J=15.6 Hz, 2H), 2.81 (Ab.sub.q, J=15.9 Hz, 2H), 1.89-2.05 (m, 1H), 1.69-1.88 (m, 1H). LC-MS: m/z=324 (MH-C.sub.6H.sub.8O.sub.7).sup.+

EXAMPLE 5

3-((3R,4R)-4-d3-Methyl-3-(d3-methyl(2-d1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2,2,3,4,5,5,6,6-d8-piperidin-1-yl)-3-oxopro-panenitrile mono citrate salt (CP-690550-d15 citrate salt)

(129) ##STR00075##
Step 1

(130) ##STR00076##

Ethyl 2-(benzylamino)acetate

(131) A solution of diisopropylethylamine (155 g, 1.2 mol) and benzylamine (96 g, 0.9 mol) was added dropwise to a solution of ethyl bromoacetate (100 g, 0.6 mol) in dioxane (1000 mL). The resulting suspension was heated at about 90 C. for about 5 hours, and then was cooled to ambient temperature. Standard extractive workup with ethyl acetate afforded the title product as a yellow oil (90 g; yield=80%). LC-MS: m/z=194 (M+H).sup.+.

(132) Step 2

(133) ##STR00077##

Ethyl 4-(benzyl(2-ethoxy-2-oxoethyl)amino)-4-oxobutanoate

(134) Potassium carbonate (110.4 g, 0.97 mol) was added in one portion to a solution of ethyl 2-(benzylamino)acetate (78 g, 0.4 mol) in tetrahydrofuran (500 mL) and water (200 mL). Ethyl 4-chloro-4-oxobutanoate (72.7 g, 0.485 mol) in anhydrous tetrahydrofuran (200 mL) was then added dropwise over a period of 1 hour to the mixture. The mixture was filtered, and the filtrate was washed with ethyl acetate. After the solvent was removed by evaporation, standard extractive workup with ethyl acetate (100 mL) to afford the title product as a yellow oil (110 g; yield=80%). LC-MS: m/z=322 (M+H).sup.+.

(135) Step 3

(136) ##STR00078##

Ethyl 1-benzyl-5-hydroxy-2-oxo-1,2,3,6-tetrahydropyridine-4-carboxylate

(137) Ethyl 4-(benzyl(2-ethoxy-2-oxoethyl)amino)-4-oxobutanoate (123.2 g, 0.4 mol) in ethanol (37 g, 0.8 mol) and dioxane (200 ml) was added dropwise to a suspension of sodium (18.4 g, 0.8 mol) in dioxane (500 mL). The resulting mixture was heated at reflux until the sodium metal was no longer visible. After cooling the mixture to ambient temperature, acetic acid (48 g, 0.8 mol) was added. Standard extractive workup with ethyl acetate, gave a crude product that was purified by re-crystallization from ether/acetone to afford the title product as a yellow solid (40 g; yield=40%). .sup.1H NMR (300 MHz, CD.sub.3OD) : 11.81 (s, 1H), 7.19-7.41 (m, 5H), 4.65 (s, 2H), 4.25 (q, J=7.2 Hz, 2H), 3.91 (t, J=3 Hz, 2H), 3.27 (t, J=3 Hz, 2H), 1.32 (t, J=7.2 Hz, 3H). LC-MS: m/z=276 (M+H).sup.+.

(138) Step 4

(139) ##STR00079##

(140) Benzyl 1-benzyl-5-hydroxy-2-oxo-1,2,3,6-tetrahydropyridine-4-carboxylate

(141) A solution of 1-benzyl-5-hydroxy-2-oxo-1, 2, 3, 6-tetrahydropyridine-4-carboxylate 4 (14 g, 50.9 mmol) in benzyl alcohol (27.5 g, 255 mmol) was heated at about 170 C. for about 16 hours. The solvent was removed in vacuo, and the resulting residue was re-crystallized from ether to give the title product as a yellow solid (14 g; yield=85%). LC-MS: m/z=338 (M+H).sup.+.

(142) Step 5

(143) ##STR00080##

Benzyl 1-benzyl-4-trideuteromethyl-2,5-dioxopiperidine-4-carboxylate

(144) A mixture of benzyl 1-benzyl-5-hydroxy-2-oxo-1,2,3,6-tetrahydropyridine-4-carboxylate 5 (13.5 g, 40 mmol), d.sub.3-iodomethane (8.7 g, 60 mmol), potassium carbonate (16.6 g, 120 mmol) and acetone (60 mL) was heated at reflux for about 3 hours. The mixture was filtered, and the resulting filtrate was concentrated in vacuo. Standard extractive workup with ethyl acetate gave a crude residue that was then purified by re-crystallization from ether/acetone to afford the title product as a light yellow solid (11.3 g; yield=80%). LC-MS: m/z=355 (M+H).sup.+.

(145) Step 6

(146) ##STR00081##

1-Benzyl-4-d3-methylpiperidine-2, 5-dione

(147) Hydrogen gas was introduced to a suspension of 1-benzyl-4-d.sub.3-methyl-2,5-dioxopiperidine-4-carboxylate (12.5 g, 35.3 mmol), 10% palladium on carbon (2 g), and ethyl acetate (100 mL). The mixture was heated at about 50 C. for about 16 hours. The mixture was then filtered through a Celite pad, and the filtrate was washed with ethyl acetate. The filtrate was heated at reflux for about 3 hours, and then the solvent was removed by evaporation in vacuo. The resulting residue was purified by silica gel column (petroleum ether/ethyl acetate) to give the title product (7 g; yield=90%). LC-MS: m/z=221 (M+H).sup.+.

(148) Step 7

(149) ##STR00082##

1-Benzyl-5,5-dimethoxy-4-d3-methylpiperidin-2-one

(150) A solution of methyl orthoformate (10 mL) and 4-methylbenzenesulfonic acid (0.5 g) in methanol (20 mL) was added dropwise to a solution of 1-benzyl-4-trideuteromethylpiperidine-2, 5-dione (7 g, 31.8 mmol) in methanol (50 mL). The resulting mixture was heated at reflux for about 16 hours and then cooled to ambient temperature. After adding triethylamine (2 ml), standard extractive workup with ethyl acetate afforded a crude residue that was then purified by silica gel column chromatography to give the title product as a yellow oil (7.8 g; yield=90%). LC-MS: m/z=267 (M+H).sup.+.

(151) Step 8

(152) ##STR00083##

1-Benzyl-5,5-dimethoxy-4-d3-methyl-3,3-d2-piperidin-2-one

(153) A mixture of 1-benzyl-5,5-dimethoxy-4-d.sub.3-methylpiperidin-2-one (4 g, 15 mmol), d.sub.4-methanol (10 mL) and 30% d.sub.1-sodium hydroxide (50 mL) was heated at about 50 C. until reaction completion, as measured by LC-MS. The mixture was cooled to ambient temperature, and deuterium oxide (25 mL) was then added. Standard extractive workup with ethyl acetate gave the title product as a yellow oil (3.3 g; yield=80%). LC-MS: m/z=269 (M+H).sup.+.

(154) Step 9

(155) ##STR00084##

1-Benzyl-4-d3-methyl-3,3,4,6,6-d5-piperidine-2,5-dione

(156) A mixture of 1-benzyl-5,5-dimethoxy-4-d.sub.3-methyl-3,3-d.sub.2-piperidin-2-one (8 g, 29.8 mmol) in 1N deuterochloric acid (in deuterium oxide) (200 mL) was heated at about 80 C. for about 16 hours. The mixture was cooled to ambient temperature, and then 2N d.sub.1-sodium hydroxide (in deuterium oxide) (110 mL) was added. The mixture was extracted with ethyl acetate, dried, and evaporated in vacuo. The resulting residue was purified by silica gel column chromatography to give the title product (4.7 g; yield=60%). LC-MS: m/z=226/244 (M+H).sup.+.

(157) Step 10

(158) ##STR00085##

1-Benzyl-4-d3-methyl-5-(d3-methylamino)-3,3,4,5,6,6-d6-piperidin-2-one

(159) At about 0 C., sodium d.sub.3-methoxide (0.9 g, 16 mmol) was added to a suspension of d.sub.5-methylamine deuterium chloride (1.2 g, 16 mmol) in tetrahydrofuran (10 mL). After 30 minutes, d.sub.4-acetic acid (1.1 g, 16 mmol) was injected into the mixture using a syringe. The resulting mixture was then stirred at ambient temperature for about 30 minutes. After replacing the atmosphere with nitrogen, 1-benzyl-4-d.sub.3-methyl-3,3,4,6,6,-d.sub.5-piperidine-2,5-dione (3 g, 13.3 mmol) in tetrahydrofuran (20 mL) was then added dropwise. The mixture was stirred for about 16 hours, and then sodium triacetoxy borodeuteride (7.4 g, 32 mmol) was added. The mixture was stirred at ambient temperature for about 5 hours, and then 5% d.sub.1-sodium hydroxide (50 mL) was added. Following standard extractive workup with ethyl acetate, the crude residue was purified by silica gel column chromatography to give the title product (1.2 g; yield=37%). LC-MS: m/z=245 (M+H).sup.+.

(160) Step 11

(161) ##STR00086##

(1-Benzyl-4-d3-methyl-2,2,3,4,5,5,6,6-d8-piperidin-3-yl)-d3-methyl-amine

(162) 1-Benzyl-4-d.sub.3-methyl-5-(d.sub.3-methylamino)-3,3,4,5,6,6-d.sub.6-piperidin-2-one (1.0 g, 4.1 mmol) in tetrahydrofuran (5 mL) was added dropwise to a suspension of lithium aluminum deuteride (860 mg, 20.5 mmol) in tetrahydrofuran (20 mL). The mixture was stirred at ambient temperature for about 1 hour. After cooling the mixture to about 10 C., the mixture was poured into 10% sodium hydroxide (5 mL) containing ice. After filtering, the filtrate was concentrated in vacuo, and extracted with ethyl acetate. The organic phases were combined, washed with brine, dried, and evaporated in vacuo, to give the title product as a yellow solid (1.0 g; yield=85%). LC-MS: m/z=233 (M+H).sup.+.

(163) Step 12

(164) ##STR00087##

N-(1-Benzyl-4-d3-methyl-2,2,3,4,5,5,6,6-d8-piperidin-3-yl)-2-chloro-N-d3-methyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine

(165) The procedure of Example 2, Step 3 was followed, but substituting (1-benzyl-4-d.sub.3-methyl-2,2,3,4,5,5,6,6-d.sub.8-piperidin-3-yl)-d.sub.3-methyl-amine for (1-benzyl-4-methyl-piperidin-3-yl)-d.sub.3-methyl-amine. The title product was isolated as a light yellow solid (1.4 g, yield=90%). LC-MS: m/z=538 (M+H).sup.+.

(166) Step 13

(167) ##STR00088##

tert-Butyl 3-((2-d1-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)(d3-methyl)amino)-4-d3-methyl-2, 2, 3, 4, 5, 5, 6, 6-d8-piperidine-1-carboxylate

(168) The procedure of Example 2, Step 4 was followed, but substituting N-(1-benzyl-4-d.sub.3-methyl-2,2,3,4,5,5,6,6-d.sub.8-piperidin-3-yl)-2-chloro-N-d.sub.3-methyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine for N-(1-benzyl-4-methylpiperidin-3-yl)-2-chloro-N-d.sub.3-methyl-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-amine. The title product was isolated as a solid. LC-MS: m/z=515 (M+H).sup.+.

(169) Step 14

(170) ##STR00089##

tert-Butyl 4-d3-methyl-3-(d3-methyl(2-d1-7H-pyrrolo [2,3-d]pyrimidin-4-yl)amino)-2, 2, 3, 4, 5, 5, 6, 6-d8-piperidine-1-carboxylate

(171) The procedure of Example 2, Step 5 was followed, but substituting tert-butyl 3-((2-d.sub.1-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)(d.sub.3-methyl)amino)-4-d.sub.3-methyl-2, 2, 3, 4, 5, 5, 6, 6-d.sub.8-piperidine-1-carboxylate for tert-butyl 4-methyl-3-(d.sub.3-methyl(2-d.sub.3-7-tosyl-7H-pyrrolo [2,3-d]pyrimidin-4-yl)amino)piperidine-1-carboxylate. The title product was isolated as a foamy solid (130 mg; yield=90%). LC-MS: m/z=361 (M+H).sup.+.

(172) Step 15

(173) ##STR00090##

(3R,4R)-tert-Butyl 3-((2-d1-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)(d3-methyl)amino-4-d3-methyl-2,2,3, 4, 5, 5,6,6-d8-piperidine-1-carboxylate

(174) The enantiomer was isolated from tert-utbyl 4-d.sub.3-methyl-3-(d.sub.3-methyl(2-d.sub.1-7H-pyrrolo [2,3-d]pyrimidin-4-yl)amino)-2, 2, 3, 4, 5, 5, 6, 6-d.sub.8-piperidine-1-carboxylate by chiral resolution using chrial-prep-HPLC with the following conditions: column: Chiralpak IC225 cm (Waters 2767-1), 5 umChiral-P(IC)001IC00CJ-LD016; mobile phase: hexane/isopropyl alcohol (85:15); detector: UV 254 nm. Retention time of desired enantiomer: 12.13 minutes, undesired enantiomer retention time: 15.15 minutes. ee %>99.8%. The title product was isolated as a yellow solid (0.1 g; yield=35%). LC-MS: m/z=361 (M+H).sup.+.

(175) Step 16

(176) ##STR00091##

N-d3-Methyl-N-((3R,4R)-4-d3-methyl2, 2, 3, 4, 5, 5, 6, 6-d8-piperidin-3-yl)-2-d1-7H-pyrrolo[2,3-d]pyrimidin-4-amine deuterochloride

(177) The procedure of Example 2, Step 7 was followed, but substituting (3R,4R)-tert-butyl 3-((2-d.sub.1-7-tosyl-7H-pyrrolo[2,3-d]pyrimidin-4-yl)(d.sub.3-methyl)amino)-4-d.sub.3-methyl-2, 2, 3, 4, 5, 5, 6, 6-d.sub.8-piperidine-1-carboxylate for 3-((3R,4R)-4-methyl-3-[d.sub.3-methyl-(2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)-amino]-piperidine)-1-carboxylic acid tert-butyl ester. The title product was isolated as a crude residue which was used in the next step without any further purification. LC-MS: m/z=261 (M+H).sup.+.

(178) Step 17

(179) ##STR00092##

3-((3R,4R)-4-d3-Methyl-3-(d3-methyl(2-d1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2, 2, 3, 4, 5, 5, 6, 6-d8-piperidin-1-yl)-3-oxopropanenitrile (CP-690550-d

(180) The procedure of Example 1, Step 11 was followed but substituting N-d.sub.3-methyl-N-((3R,4R)-4-d.sub.3-methyl-2, 2, 3, 4, 5, 5, 6, 6-d.sub.8-piperidin-3-yl)-2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-amine deuterochloride for N-methyl-N-((3R,4R)-4-methylpiperidin-3-yl)-7H-pyrrolo[2,3-d]pyrimidin-4-amine. The title product was isolated as a light yellow solid (50 mg; yield=63%). LC-MS: m/z=328 (M+H).sup.+.

(181) Step 18

(182) ##STR00093##

3-((3R,4R)-4-d3-Methyl-3-(d3-methyl(2-d1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2, 2, 3, 4, 5, 5, 6, 6-d8-piperidin-1-yl)-3-oxopropanenitrile mono citrate (CP-690550-d15 citrate salt)

(183) The procedure of Example 1, Step 12 was followed but substituting 3-((3R,4R)-4-d.sub.3-methyl-3-(d.sub.3-methyl(2-d.sub.1-7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2, 2, 3, 4, 5, 5, 6, 6-d.sub.8-piperidin-1-yl)-3-oxopropanenitrile for 3-((3R,4R)-4-methyl-3-(methyl(7H-pyrrolo[2,3-d]pyrimidin-4-yl)amino)piperidin-1-yl)-3-oxopropanenitrile. The title product was isolated as a white solid (54 mg; yield=90%). .sup.1H NMR (300 MHz, CD.sub.3OD) : 7.35 (s, 1H), 6.89 (d, J=2.7 Hz, 1H), 3.91-4.08 (m, 2H), 2.94 (Ab.sub.q, J=15.6 Hz, 2H), 2.81 (Ab.sub.q, J=15.9 Hz, 2H). LC-MS: m/z=328 (MH-C.sub.6H.sub.8O.sub.7).sup.+.

(184) The following compounds can generally be made using the methods described above. It is expected that these compounds when made will have activity similar to those described in the examples above.

(185) ##STR00094## ##STR00095## ##STR00096## ##STR00097## ##STR00098## ##STR00099## ##STR00100## ##STR00101## ##STR00102## ##STR00103## ##STR00104## ##STR00105## ##STR00106## ##STR00107## ##STR00108## ##STR00109## ##STR00110## ##STR00111## ##STR00112## ##STR00113## ##STR00114## ##STR00115## ##STR00116## ##STR00117## ##STR00118## ##STR00119## ##STR00120## ##STR00121## ##STR00122## ##STR00123## ##STR00124## ##STR00125## ##STR00126## ##STR00127## ##STR00128## ##STR00129## ##STR00130## ##STR00131##

(186) ##STR00132## ##STR00133## ##STR00134## ##STR00135## ##STR00136## ##STR00137## ##STR00138## ##STR00139## ##STR00140## ##STR00141## ##STR00142## ##STR00143## ##STR00144## ##STR00145## ##STR00146## ##STR00147## ##STR00148## ##STR00149## ##STR00150## ##STR00151## ##STR00152## ##STR00153## ##STR00154## ##STR00155## ##STR00156## ##STR00157## ##STR00158## ##STR00159## ##STR00160## ##STR00161## ##STR00162## ##STR00163## ##STR00164## ##STR00165## ##STR00166## ##STR00167## ##STR00168## ##STR00169##

(187) Changes in the metabolic properties of the compounds disclosed herein as compared to their non-isotopically enriched analogs can be shown using the following assays. Compounds listed above which have not yet been made and/or tested are predicted to have changed metabolic properties as shown by one or more of these assays as well.

Biological Activity Assays

(188) In vitro Human Liver Microsomal Stability (HLM) Assay

(189) Liver microsomal stability assays were conducted with 4 mg per mL liver microsome protein with an NADPH-generating system (8.8 mM NADPH, 102.4 mM glucose 6-phosphate, 24 units per mL glucose 6-phosphate dehydrogenase and 13.2 mM magnesium chloride) in 2% sodium bicarbonate. Test compounds were prepared as solutions in 20% acetonitrile-water and added to the assay mixture (final assay concentration 5 microgram per mL) and incubated at 37 C. Final concentration of acetonitrile in the assay should be <1%. Aliquots (50 L) were taken out at times 0, 30, 60, 90, and 120 minutes, and diluted with ice cold acetonitrile (200 L) to stop the reactions. Samples were centrifuged at 12,000 RPM for 10 minutes to precipitate proteins. Supernatants were transferred to microcentrifuge tubes and stored for LC/MS/MS analysis of the degradation half-life of the test compounds. It has thus been found that certain isotopically enriched compounds disclosed herein that have been tested in this assay showed an increased degradation half-life as compared to the non-isotopically enriched drug. In certain embodiments, the increase in degradation half-life is at least 5%; at least 10%; at least 15%; at least 20%; at least 30%; at least 40%; at least 50%; at least 60%; at least 70%; at least 80%; at least 90%; or at least 100%.

(190) In vitro Metabolism Using Human Cytochrome P.sub.450 Enzymes

(191) The cytochrome P.sub.450 enzymes are expressed from the corresponding human cDNA using a baculovirus expression system (BD Biosciences, San Jose, Calif.). A 0.25 milliliter reaction mixture containing 0.8 milligrams per milliliter protein, 1.3 millimolar NADP.sup.+, 3.3 millimolar glucose-6-phosphate, 0.4 U/mL glucose-6-phosphate dehydrogenase, 3.3 millimolar magnesium chloride and 0.2 millimolar of a compound as disclosed herein, the corresponding non-isotopically enriched compound or standard or control in 100 millimolar potassium phosphate (pH 7.4) is incubated at 37 C. for 20 minutes. After incubation, the reaction is stopped by the addition of an appropriate solvent (e.g., acetonitrile, 20% trichloroacetic acid, 94% acetonitrile/6% glacial acetic acid, 70% perchloric acid, 94% acetonitrile/6% glacial acetic acid) and centrifuged (10,000 g) for 3 minutes. The supernatant is analyzed by HPLC/MS/MS.

(192) TABLE-US-00001 Cytochrome P.sub.450 Standard CYP1A2 Phenacetin CYP2A6 Coumarin CYP2B6 [.sup.13C]-(S)-mephenytoin CYP2C8 Paclitaxel CYP2C9 Diclofenac CYP2C19 [.sup.13C]-(S)-mephenytoin CYP2D6 (+/)-Bufuralol CYP2E1 Chlorzoxazone CYP3A4 Testosterone CYP4A [.sup.13C]-Lauric acid
Monoamine Oxidase A Inhibition and Oxidative Turnover

(193) The procedure is carried out using the methods described by Weyler et al., Journal of Biological Chemistry 1985, 260, 13199-13207, which is hereby incorporated by reference in its entirety. Monoamine oxidase A activity is measured spectrophotometrically by monitoring the increase in absorbance at 314 nm on oxidation of kynuramine with formation of 4-hydroxyquinoline. The measurements are carried out, at 30 C., in 50 mM sodium phosphate buffer, pH 7.2, containing 0.2% Triton X-100 (monoamine oxidase assay buffer), plus 1 mM kynuramine, and the desired amount of enzyme in 1 mL total volume.

(194) Monooamine Oxidase B Inhibition and Oxidative Turnover

(195) The procedure is carried out as described in Uebelhack, Pharmacopsychiatry 1998, 31(5), 187-192, which is hereby incorporated by reference in its entirety.

(196) Detecting CP-690550 and its metabolites by LC-MS

(197) The procedure is carried out as described in Lawendy et al., J Clin Pharmacol 2009, 49, 423-429, which is hereby incorporated by reference in its entirety.

(198) Quantifying CP-690550 in whole blood by LC-MS

(199) The procedure is carried out as described in Paniagua et al., Therapeutic Drug Monitoring 2005, 27(5), 608-616, which is hereby incorporated by reference in its entirety.

(200) Janus Kinase 3 Enzymatic Assay

(201) The procedure is carried out as described in U.S. Pat. No. 6,627,754, which is hereby incorporated by reference in its entirety.

(202) Janus Kinase 3 Enzymatic Assay

(203) The procedure is carried out as described in WO 2003/048162, which is hereby incorporated by reference in its entirety.

(204) Inhibition of Human IL-2 Dependent T-Cell Blast Proliferation

(205) The procedure is carried out as described in WO 2003/048162, which is hereby incorporated by reference in its entirety.

(206) From the foregoing description, one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various usages and conditions.