SULFONAMIDE-SUBSTITUTED CYANOPYRROLIDINES WITH ACTIVITY AS DUB INHIBITORS

Abstract

The present invention relates to a class of sulfonamide-substituted cyanopyrrolidines of Formula (Ia) and (Ib) with activity as inhibitors of deubiquitilating enzymes, in particular, ubiquitin C-terminal hydrolase L1 (UCHL1) and ubiquitin C-terminal hydrolase 30 or ubiquitin specific peptidase 30 (USP30), having utility in a variety of therapeutic areas including cancer and conditions involving mitochondrial dysfunction: (Formulae (Ia), (Ib)).

##STR00001##

Claims

1. A compound of formula (I), which is selected from (Ia) and (lb): ##STR00063## a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer, wherein: m is 0 to 4; n is 0 or 1; each R.sup.1 is independently selected from halo, cyano, hydroxy, (C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy, halo(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkoxy, and (C.sub.1-C.sub.6)alkoxy(C.sub.1-C.sub.6)alkyl; R.sup.2 and R.sup.3 are selected from hydrogen, (C.sub.1-C.sub.6)alkyl, and (C.sub.1-C.sub.6)alkoxy(C.sub.1-C.sub.6)alkyl; or where R.sup.1 and NR.sup.2 are situated on adjacent ring atoms, R.sup.1 together with R.sup.2 may form a 5 to 6-membered heterocyclic ring containing 1 to 2 heteroatoms independently selected from N, O and S, at least one of which is N; L.sup.1 is selected from a covalent bond, (C.sub.1-C.sub.4)alkylene, and (C.sub.2-C.sub.4)alkenylene; L.sup.2 is selected from a covalent bond, (C.sub.1-C.sub.4)alkylene, (C.sub.2-C.sub.4)alkenylene, and (C.sub.0-C.sub.3)alkylene-X(C.sub.0-C.sub.3)alkylene; X is selected from O, S, SO, SO.sub.2, NR.sup.4, NR.sup.4C(O), C(O)NR.sup.4, NR.sup.4C(O)NR.sup.5, C(O), C(O)O, OC(O), OC(O)O, SO.sub.2NR.sup.4, NR.sup.4SO.sub.2, and NR.sup.4SO.sub.2NR.sup.5; R.sup.4 and R.sup.5 are each independently selected from hydrogen, (C.sub.1-C.sub.6)alkyl, and (C.sub.1-C.sub.6)alkoxy(C.sub.1-C.sub.6)alkyl; group A is selected from a 3 to 10-membered carbocyclic ring, and a 3 to 10-membered heterocyclic ring comprising 1 to 4 heteroatoms independently selected from N, O, and S; with the proviso that for the compound of formula (Ia) when L.sup.1 is a covalent bond, A is linked to the sulfonamide via a ring C-atom; or A-L.sup.1-NR.sup.3 may optionally form a 3 to 10-membered heterocyclic ring comprising 1 to 4 heteroatoms independently selected from N, O, and S, at least one of which is N; group B is selected from a 3 to 10-membered carbocyclic ring, and a 3 to 10-membered heterocyclic ring comprising 1 to 4 heteroatoms independently selected from N, O, and S; and each carbocyclic and heterocyclic ring may be optionally substituted with 1 to 4 substituents independently selected from halo, cyano, hydroxy, oxo, (C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy, (C.sub.1-C.sub.6)alkoxy(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkyl, halo(C.sub.1-C.sub.6)alkoxy, NH(C.sub.1-C.sub.6)alkyl, N((C.sub.1-C.sub.6)alkyl).sub.2, C(O)NH(C.sub.1-C.sub.6)alkyl, C(O)N((C.sub.1-C.sub.6)alkyl).sub.2, NHC(O)(C.sub.1-C.sub.6)alkyl, N(C.sub.1-C.sub.6)alkyl)C(O)(C.sub.1-C.sub.6)alkyl), C(O)(C.sub.1-C.sub.6)alkyl, C(O)O(C.sub.1-C.sub.6)alkyl, CO.sub.2H, CONH.sub.2, SO.sub.2NH(C.sub.1-C.sub.6)alkyl, and SO.sub.2N((C.sub.1-C.sub.6)alkyl).sub.2.

2. The compound according claim 1, wherein m is 0, 1 or 2.

3. The compound according to claim 1, wherein each R.sup.1 is independently selected from fluoro, cyano, methyl, methoxy, and methoxymethyl; R.sup.2 and R.sup.3 are selected from hydrogen and methyl; or where R.sup.1 and NR.sup.2 are situated on adjacent ring atoms, R.sup.1 together with R.sup.2 may form a morpholine, piperidine, or pyrrolidine ring.

4. The compound compound according to claim 1, wherein L.sup.1 is selected from a covalent bond, methylene, and ethylene.

5. The compound according to claim 1, wherein L.sup.2 is selected from a covalent bond, an oxygen atom, methylene, OCH.sub.2, and NHC(O).

6. The compound according to claim 1, wherein group A is selected from indanyl, phenyl, tetrahydrofuranyl, tetrahydropyranyl, tetralinyl, benzothiazolyl, imidazolyl, isoxazolyl, piperidinyl, pyrazolyl, pyridyl, pyrimidinyl, thiazolyl, 1,2,4-triazolyl, and quinolinyl; or A-L.sup.1-NR.sup.3 may form a ring selected from azetidinyl, isoindolinyl, piperazinyl, piperidinyl, tetrahydroisoquinolinyl, and 4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinyl.

7. The compound according to claim 1, wherein group B is selected from phenyl, oxazolyl, piperazinyl, piperidinyl, pyrazolyl, pyridyl, pyrimidinyl, pyrrolidinyl, 1,2-thiazolidinyl, and thiazolyl.

8. The compound according to claim 1, each carbocyclic and heterocyclic ring may be optionally substituted with 1 to 2 substituents independently selected from halo, cyano, hydroxy, oxo, (C.sub.1-C.sub.6)alkyl, (C.sub.1-C.sub.6)alkoxy, CF.sub.3, OCF.sub.3, and NHC(O)(C.sub.1-C.sub.6)alkyl.

9. The compound according to claim 8, wherein each carbocyclic and heterocyclic ring may be optionally substituted with 1 to 2 substituents independently selected from chloro, fluoro, cyano, hydroxy, oxo, methyl, isopropyl, methoxy, CF.sub.3, OCF.sub.3, NHC(O)isobutyl.

10. The compound according to claim 1, selected from the group consisting of: N-(1-cyanopyrrolidin-3-yl)-[1,1-biphenyl]-2-sulfonamide; N-(1-cyanopyrrolidin-3-yl)-[1,1-biphenyl]-4-sulfonamide; (R)N-(1-cyanopyrrolidin-3-yl)-[1,1-biphenyl]-4-sulfonamide; (S)N-(1-cyanopyrrolidin-3-yl)-[1,1-biphenyl]-4-sulfonamide; N-(1-cyanopyrrolidin-3-yl)benzenesulfonamide; (S)N-(1-cyanopyrrolidin-3-yl)benzenesulfonamide; (R)N-(1-cyanopyrrolidin-3-yl)benzenesulfonamide; (4-(benzyloxy)-N-(1-cyanopyrrolidin-3-yl)benzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-(pyrimidin-5-yl)benzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-(4-methylpiperazin-1-yl)benzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-5-phenylpyridine-2-sulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-fluoro-[1,1-biphenyl]-4-sulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-(pyridin-4-yl)benzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-(piperidin-1-yl)benzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-(1-methyl-1H-pyrazol-4-yl)benzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-6-phenoxypyridine-3-sulfonamide; N-(1-cyanopyrrolidin-3-yl)-5-fluoro-2-methylbenzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-isopropylbenzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-((5-(trifluoromethyl)pyridin-2-yl)oxy)benzenesulfonamide; (R)N-(1-cyanopyrrolidin-3-yl)-N-methyl-4-(pyridin-3-yl)benzenesulfonamide; (3aR,6aR)-1-((4-(pyridin-3-yl)phenyl)sulfonyl)hexahydropyrrolo[3,4-b]pyrrole-5(1H)-carbonitrile; rac-(4aR,7aR)-4-tosylhexahydropyrrolo[3,4-b][1,4]oxazine-6(2H)-carbonitrile; rac-(4aR,7aS)-4-((4-methylbenzyl)sulfonyl)hexahydropyrrolo[3,4-b][1,4]oxazine-6(2H)-carbonitrile; N-(5-(N-(1-cyanopyrrolidin-3-yl)sulfamoyl)pyridin-2-yl)-1-methyl-1H-pyrazole-5-carb oxamide; N-(1-cyanopyrrolidin-3-yl)-N-methyl-4-((5-(trifluoromethyl)pyridin-2-yl)oxy)benzenesulfonamide; N-([1,1-biphenyl]-4-yl)-1-cyanopyrrolidine-3-sulfonamide; 3-((4-(4-fluorophenyl)piperazin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; (3-((4-(pyridin-2-yl)piperazin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; 3-((4-(pyrimidin-2-yl)piperazin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; 3-((4-(4-chlorophenyl)piperidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; (S)-3-((4-(4-chlorophenyl)piperidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; (R)-3-((4-(4-chlorophenyl)piperidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; 3-((4-benzylpiperidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; 3-((3-(4-chlorophenyl)azetidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; (S)-3-((3-(4-chlorophenyl)azetidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; (R)-3-((3-(4-chlorophenyl)azetidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; and 3-((3-phenoxyazetidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer.

11. A method of treatment of a disorder or condition where inhibition of UCHL1 or USP30 is known, or can be shown, to produce a beneficial effect, in a mammal, comprising administering to said mammal a therapeutically effective amount of a compound according to claim 1, a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer.

12. (canceled)

13. The method according to claim 11, wherein the disorder or condition is selected from cancer, neurodegenerative disorders, inflammation, viral infection, bacterial infection, metabolic disorders, mitochondrial dysfunction, and fibrosis; and preferably: cancer (for example, breast, ovarian, prostate, lung, kidney, gastric, colon, testicular, head and neck, pancreas, brain, melanoma, bone or other cancers of tissue organs and cancers of the blood cells, such as lymphoma and leukaemia, multiple myeloma, colorectal cancer, and non-small cell lung carcinoma), Parkinson's disease (PD), Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Huntington's disease, ischemia, stroke, dementia with Lewy bodies, and frontotemporal dementia; PD related to mutations in -synuclein, parkin and PINK1, autosomal recessive juvenile Parkinson's disease (AR-JP) where parkin is mutated; chronic obstructive pulmonary disease (COPD), inflammation, viral infections, MERS, SARS, bacterial infections, TB, metabolic disorders; multiple sclerosis (MS), mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS) syndrome; Leber's hereditary optic neuropathy (LHON); neuropathy, ataxia, retinitis pigmentosa-maternally inherited Leigh syndrome (NARP-MILS); Danon disease; diabetes; diabetic nephropathy; metabolic disorders; heart failure; ischemic heart disease leading to myocardial infarction; psychiatric diseases, schizophrenia; multiple sulfatase deficiency (MSD); mucolipidosis II (ML II); mucolipidosis III (ML III); mucolipidosis IV (ML IV); GM1-gangliosidosis (GM1); neuronal ceroid-lipofuscinoses (NCL1); Alpers disease; Barth syndrome; Beta-oxidation defects; carnitine-acyl-carnitine deficiency; carnitine deficiency; creatine deficiency syndromes; co-enzyme Q10 deficiency; complex I deficiency; complex II deficiency; complex III deficiency; complex IV deficiency; complex V deficiency; COX deficiency; chronic progressive external ophthalmoplegia syndrome (CPEO); CPT I deficiency; CPT II deficiency; glutaric aciduria type II; Kearns-Sayre syndrome; lactic acidosis; long-chain acyl-CoA dehydrogenase deficiency (LCHAD); Leigh disease or syndrome; lethal infantile cardiomyopathy (LIC); Luft disease; glutaric aciduria type II; medium-chain acyl-CoA dehydrogenase deficiency (MCAD); myoclonic epilepsy and ragged-red fiber (MERRF) syndrome; mitochondrial cytopathy; mitochondrial recessive ataxia syndrome; mitochondrial DNA depletion syndrome; myoneurogastointestinal disorder and encephalopathy; Pearson syndrome; pyruvate dehydrogenase deficiency; pyruvate carboxylase deficiency; POLG mutations; medium/short-chain 3-hydroxyacyl-CoA dehydrogenase (M/SCHAD) deficiency; and very long-chain acyl-CoA dehydrogenase (VLCAD) deficiency; fibrosis or a fibrotic disorder associated with the accumulation of extracellular matrix constituents that occurs following trauma, inflammation, tissue repair, immunological reactions, cellular hyperplasia, and neoplasia; fibrosis or a fibrotic disorder associated with major organ diseases, fibroproliferative disorders, and scarring associated with trauma; fibrosis or a fibrotic disorder associated with interstitial lung disease (ILD), liver cirrhosis, non-alcoholic fatty liver disease (NAFLD), non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), kidney disease, heart or vascular disease, diseases of the eye, systemic and local scleroderma, keloids, hypertrophic scars, atherosclerosis, restenosis, Dupuytren's contracture, surgical complications, chemotherapeutics drug-induced fibrosis, radiation-induced fibrosis, accidental injury and burns, retroperitoneal fibrosis, and peritoneal fibrosis/peritoneal scarring; fibrosis associated with interstitial lung disease is selected from sarcoidosis, silicosis, drug reactions, infections, collagen vascular diseases, rheumatoid arthritis, systemic sclerosis, scleroderma, pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), usual interstitial pneumonitis (UIP), interstitial lung disease, cryptogenic fibrosing alveolitis (CFA), bronchiolitis obliterans, and bronchiectasis; and fibrosis associated with liver cirrhosis is selected from cirrhosis associated with viral hepatitis, schistosomiasis and chronic alcoholism.

14. The compound of formula (Ia) according to claim 1, a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer, wherein: m is 1 to 4; one of the R.sup.1 groups and NR.sup.2 are situated on adjacent ring atoms, and said R.sup.1 together with R.sup.2 forms a 5 to 6-membered heterocyclic ring containing 1 to 2 heteroatoms independently selected from N, O and S, at least one of which is N.

15. The compound according to claim 14, a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer, wherein: m is 1; L.sup.1 is selected from a covalent bond, methylene, and ethylene; L.sup.2 is a covalent bond; group A is phenyl; group B is pyridyl; and each phenyl and pyridyl ring may be optionally substituted with 1 to 2 substituents independently selected from chloro, fluoro, cyano, hydroxy, methyl, isopropyl, methoxy, CF.sub.3, OCF.sub.3, NHC(O)isobutyl.

16. The compound of formula (Ia) according to claim 1, a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer, wherein: n is 0; group A is a 3 to 10-membered heterocyclic ring comprising 1 to 4 heteroatoms independently selected from N, O, and S; with the proviso that for the compound of formula (Ia) when L.sup.1 is a covalent bond, A is linked to the sulfonamide via a ring C-atom; with the proviso: (a) when m is 0; R.sup.2 is selected from (C.sub.1-C.sub.6)alkyl, and (C.sub.1-C.sub.6)alkoxy(C.sub.1-C.sub.6)alkyl.

17. The compound of formula (Ia) according to claim 1, a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer, wherein: m is 0; n is 0; R.sup.2 is hydrogen; and each carbocyclic and heterocyclic ring is substituted with, at least, 1 to 2 substituents independently selected from cyano, hydroxy, oxo, CF.sub.3, OCF.sub.3, and NHC(O)isobutyl.

18. The compound of formula (Ia) according to claim 1, a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer, wherein: n is 1; and group B is a substituted 3 to 10-membered heterocyclic ring comprising 1 to 4 heteroatoms independently selected from N, O, and S.

19. The compound of formula (Ib) according to claim 1, a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer.

20. The compound according to claim 19, wherein m is 0 or 1; n is 0 or 1; each R.sup.1 is independently selected from fluoro, cyano, methyl, methoxy, and methoxymethyl; R.sup.3 is selected from hydrogen and methyl; L.sub.1 is selected from a covalent bond, methylene, and ethylene; L.sup.2 is selected from a covalent bond, an oxygen atom, and methylene; group A is selected from indanyl, phenyl, tetralinyl, benzothiazolyl, piperidinyl, pyrazolyl, pyridyl, pyrimidinyl, thiazolyl, 1,2,4-triazolyl, and quinolinyl; or A-L.sup.1-NR.sup.3 may form a ring selected from azetidinyl, isoindolinyl, piperazinyl, piperidinyl, tetrahydroisoquinolinyl, and 4,5,6,7-tetrahydropyrazolo[1,5-a]pyrazinyl; group B is selected from phenyl and pyridyl; and each carbocyclic and heterocyclic ring may be optionally substituted with 1 to 2 substituents independently selected from chloro, fluoro, hydroxy, and methyl.

21. A compound selected from the group consisting of: N-(1-cyanopyrrolidin-3-yl)-[1,1-biphenyl]-2-sulfonamide; N-(1-cyanopyrrolidin-3-yl)-[1,1-biphenyl]-4-sulfonamide; (R)N-(1-cyanopyrrolidin-3-yl)-[1,1-biphenyl]-4-sulfonamide; (S)N-(1-cyanopyrrolidin-3-yl)-[1,1-biphenyl]-4-sulfonamide; (4-(benzyl oxy)-N-(1-cyanopyrrolidin-3-yl)benzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-(pyrimidin-5-yl)benzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-(4-methylpiperazin-1-yl)benzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-5-phenylpyridine-2-sulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-(pyridin-4-yl)benzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-(piperidin-1-yl)benzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-(1-methyl-1H-pyrazol-4-yl)benzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-6-phenoxypyridine-3-sulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-isopropylbenzenesulfonamide; N-(1-cyanopyrrolidin-3-yl)-4-((5-(trifluoromethyl)pyridin-2-yl)oxy)benzenesulfonamide; (R)N-(1-cyanopyrrolidin-3-yl)-N-methyl-4-(pyridin-3-yl)benzenesulfonamide; (3aR,6aR)-1-((4-(pyridin-3-yl)phenyl)sulfonyl)hexahydropyrrolo[3,4-b]pyrrole-5(1H)-carbonitrile; rac-(4aR,7aR)-4-tosylhexahydropyrrolo[3,4-b][1,4]oxazine-6(2H)-carbonitrile; rac-(4aR,7aS)-4-((4-methylbenzyl)sulfonyl)hexahydropyrrolo[3,4-b][1,4]oxazine-6(2H)-carbonitrile; N-(5-(N-(1-cyanopyrrolidin-3-yl)sulfamoyl)pyridin-2-yl)-1-methyl-1H-pyrazole-5-carboxamide; N-(1-cyanopyrrolidin-3-yl)-N-methyl-4-((5-(trifluoromethyl)pyridin-2-yl)oxy)benzenesulfonamide; N-([1,1-biphenyl]-4-yl)-1-cyanopyrrolidine-3-sulfonamide; 3-((4-(4-fluorophenyl)piperazin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; (3-((4-(pyridin-2-yl)piperazin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; 3-((4-(pyrimidin-2-yl)piperazin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; 3-((4-(4-chlorophenyl)piperidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; (S)-3-((4-(4-chlorophenyl)piperidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; (R)-3-((4-(4-chlorophenyl)piperidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; 3-((4-benzylpiperidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; 3-((3-(4-chlorophenyl)azetidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; (S)-3-((3-(4-chlorophenyl)azetidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; (R)-3-((3-(4-chlorophenyl)azetidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; and 3-((3-phenoxyazetidin-1-yl)sulfonyl)pyrrolidine-1-carbonitrile; a tautomer thereof, or a pharmaceutically acceptable salt of said compound or tautomer.

22. A pharmaceutical composition, comprising a compound according to claim 1, or a pharmaceutically acceptable salt of said compound or tautomer, together with a pharmaceutically acceptable diluent or carrier.

23. (canceled)

24. A compound of formula: (a) (IIa) or (IIa): ##STR00064## wherein PG is a protecting group, preferably BOC or CBZ, and m, n, R.sup.1, R.sup.2, L.sup.1, L.sup.2, group A and group B are as defined in any one of claims 14 to 18, a tautomer thereof, or a salt of said compound or tautomer; or (b) (IIb) or (IIIb): ##STR00065## wherein PG is a protecting group, preferably BOC or CBZ, and m, n, R.sup.1, R.sup.3, L.sup.1, L.sup.2, group A and group B are as defined in either claim 19 or claim 20, a tautomer thereof, or a salt of said compound or tautomer.

25. A pharmaceutical composition, comprising a compound according to claim 21, or a pharmaceutically acceptable salt of said compound or tautomer, together with a pharmaceutically acceptable diluent or carrier.

Description

EXAMPLE 20

(R)N-(1-cyanopyrrolidin-3-yl)-N-methyl-4-(pyridin-3-yl)benzenesulfonamide

[0306] ##STR00037##

Step 1. tert-Butyl (R)-3-((4-bromo-N-methylphenyl)sulfonamido) pyrrolidine-1-carboxylate)

[0307] ##STR00038##

[0308] To a stirred solution of tert-butyl (R)-3-(methylamino)pyrrolidine-1-carboxylate (CAS no. 199336-83-9, Available from Combi Blocks) (0.500 g, 2.50 mmol) in THF was added TEA (0.756 g, 7.49 mmol) at 0 C. under nitrogen and stirred for 10 min at the same temperature. A solution of 4-bromobenzenesulfonyl chloride (CAS No. 98-58-8, available from Combi-blocks) (0.636 g, 2.50 mmol) in THF (1 mL) was slowly added to the reaction mixture at 0 C. The reaction mixture was stirred for 1 h at room temperature. The resulting reaction mixture was diluted with water (30 mL) and was extracted with EtOAc (320 mL). The combined organic phases were dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to yield tert-butyl (R)-3-((4-bromo-N-methylphenyl)sulfonamido)-pyrrolidine-1-carboxylate [1.00 g, 95.5% (crude)]. LCMS: Method C, 2.427 min, MS: ES+363.1, 365.1 (M56).

Step 2. tert-Butyl (R)-3-((N-methyl-4-(pyridin-3-yl)phenyl)sulfonamido)pyrrolidine-1-carboxylate

[0309] ##STR00039##

[0310] To a mixture of tert-butyl (R)-3-((4-bromo-N-methylphenyl)sulfonamido)pyrrolidine-1-carboxylate (1.0 g, 2.39 mmol) and pyridine-3-boronic acid (CAS No. 1692-25-7, available from Combi-blocks) (0.293 g, 2.39 mmol) in DMF-water (3:2; 5 mL) was added Na.sub.2CO.sub.3 (0.505 g, 4.77 mmol). Resulting mixture was degassed (by purging nitrogen through the reaction solution) for 15 to 20 min. Tetrakis(triphenylphosphine)palladium (O) (0.275 g, 0.24 mmol) was added into the reaction solution and the resulting mixture was stirred at 110 C. for 16 h. The reaction mixture was cooled to room temperature and diluted with water (110 mL) and was extracted with EtOAc (350 mL). The combined organic extracts were dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to yield tert-butyl (R)-3-((N-methyl-4-(pyridin-3-yl)phenyl)sulfonamido)pyrrolidine-1-carboxylate (0.5 g, 1.199 mmol) as crude mass. LCMS: Method C, 2.035 min, MS: ES+418.3.

Step 3. (R)N-methyl-4-(pyridin-3-yl)-N-(pyrrolidin-3-yl)benzenesulfonamide TFA salt

[0311] ##STR00040##

[0312] To a solution of (R)-3-((N-methyl-4-(pyridin-3-yl)phenyl)sulfonamido)pyrrolidine-1-carboxylate (0.4 g, 0.96 mmol) in DCM (5 mL) was added TFA (1 mL) at 0 C. and the resulting solution was stirred at room temperature for 4 h. Reaction mixture was concentrated under reduced pressure. The crude was azeotropically distilled with diethyl ether (210 mL) and dried under reduced pressure to afford (R)N-methyl-4-(pyridin-3-yl)-N-(pyrrolidin-3-yl)benzenesulfonamide TFA salt (0.4 g, quantitative). LCMS: Method C, 1.377 min, MS: ES+318.3.

Step 4. (R)N-(1-cyanopyrrolidin-3-yl)-N-methyl-4-(pyridin-3-yl)-benzenesulfonamide

[0313] To a stirred solution (R)N-methyl-4-(pyridin-3-yl)-N-(pyrrolidin-3-yl)-benzenesulfonamide TFA salt (0.400 g, 0.93 mmol) in THF (5 mL) was added K.sub.2CO.sub.3 (0.384 g, 2.78 mmol) at 0 C. CNBr (0.118 g, 1.11 mmol) was added into the reaction mixture at 0 C. The reaction mixture was stirred at room temperature for 1 h. The resulting reaction mixture was poured in to water (50 mL) and was extracted with EtOAc (350 mL). Combined organic extracts were dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The resulting residue was purified by Combi-flash Column chromatography (silica; eluted with 2.5% MeOH in DCM) yielding title compound (0.080 g, 0.23 mmol). LCMS: Method B, 3.096 min, MS: ES+343.1 [M+1]; .sup.1H NMR (400 MHz, CDCl3) ppm: 9.00 (d, J=2 Hz, 1H), 8.66 (dd, J=4.8, 1.2 Hz, 1H), 8.19 (dt, J=8.0, 1.6 Hz, 1H), 8.01 (d, J=8.8 Hz, 2H), 7.93 (d, J=8.4 Hz, 2H), 7.57-7.54 (m, 1H), 4.64-4.60 (m, 1H), 3.45-3.40 (m, 2H), 3.33-3.29 (m, 1H), 3.20-3.15 (m, 1H), 2.72 (s, 3H), 1.88-1.83 (m, 1H), 1.79-1.73 (m, 1H).

EXAMPLE 21

(3aR,6aR)-1-((4-(pyridin-3-yl)phenyl)sulfonyl)hexahydropyrrolo[3,4-b]pyrrole-5(1H)-carbonitrile

[0314] ##STR00041##

Step 1. tert-butyl rac-(3aR,6aR)-1-((4-bromophenyl)sulfonyl)hexahydropyrrolo-[3,4-b]pyrrole-5(1H)-carboxylate

[0315] ##STR00042##

[0316] To a stirred solution of 4-bromobenzene sulfonyl chloride (CAS No. 98-58-8, available from Alfa Aesar) (0.2 g, 0.78 mmol) and tert-butyl rac-(3aS,6aS)-hexahydropyrrolo[3,4-b]pyrrole-5(1H)-carboxylate (CAS No. 180975-51-3, available from Enamine) (0.166 g, 0.78 mmol) in THF (10 mL) was added K.sub.2CO.sub.3 (0.324 g, 2.35 mmol) at ambient temperature. The reaction mixture was stirred at room temperature for 8 h. The resulting reaction mixture was diluted with water (200 mL) and was extracted with diethyl ether (3100 mL).

[0317] Combined organic extracts were dried over anhydrous Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure to yield tert-butyl rac-(3aR,6aR)-1-((4-bromophenyl)sulfonyl)hexahydropyrrolo[3,4-b]pyrrole-5(1H)-carboxylate (0.350 g, Quantitative) as crude mass. LCMS: Method C, 2.667 min, MS: ES+448.3, 450.3 (M+18).

Step 2. tert-butyl (3aR,6aR)-1-((4-(pyridin-3-yl)phenyl)sulfonyl)hexahydropyrrolo-[3,4-b]pyrrole-5(1H)-carboxylate

[0318] ##STR00043##

[0319] To a mixture of tert-butyl rac-(3aR,6aR)-1-((4-bromophenyl)sulfonyl)-hexahydropyrrolo[3,4-b]pyrrole-5(1H)-carboxylate (0.350 g, 0.81 mmol), pyridine-3-boronic acid (CAS No. 1692-25-7, available from Combiblocks) (0.120 g, 0.98 mmol) and in DMF-water (4:1; 17.5 mL) was added K.sub.2CO.sub.3 (0.224 g, 1.62 mmol). The reaction mixture was degassed (by purging nitrogen through the solution) for 30 min. PdCl.sub.2(dppf) (0.060 g, 0.08 mmol) was added into the reaction mixture and the resulting mixture was stirred at 80 C. for 2 h. Reaction mixture was cooled to room temperature, diluted with saturated aqueous NaHCO.sub.3 (350 mL) and was extracted with EtOAc (3200 mL). The combined organic phase was dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The resulting residue was purified by Flash column chromatography (eluting with 3% MeOH in DCM) to yield tert-butyl rac-(3aR,6aR)-1-((4-(pyridin-3-yl)phenyl)sulfonyl)hexa-hydropyrrolo[3,4-b]pyrrole-5(1H)-carboxylate (0.350 g, 0.82 mmol). LCMS: Method C, 2.160 min, MS: ES+430.5

[0320] Step 3. rac-(3aR,6aR)-1-((4-(pyridin-3-yl)phenyl)sulfonyl)octahydropyrrolo[3,4-b]pyrrole TFA salt

##STR00044##

[0321] To a stirred solution of tert-butyl rac-(3aR,6aR)-1-((4-(pyridin-3-yl)phenyl)sulfonyl)hexahydropyrrolo[3,4-b]pyrrole-5(1H)-carboxylate (0.350 g, 0.82 mmol) in DCM (20 mL) was added TFA (2 mL) at 0 C. The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated under reduced pressure. The crude residue was further azetropically distilled with diethyl ether (310 mL) and dried under reduced pressure to afford rac-(3aR,6aR)-1-((4-(pyridin-3-yl)phenyl)sulfonyl) octahydropyrrolo[3,4-b]pyrrole TFA salt (0.210 g, quantitative) as crude mass. LCMS: Method C, 1.472 min, MS: ES+330.29

Step 4. (3aR,6aR)-1-((4-(pyridin-3-yl)phenyl)sulfonyl)hexahydropyrrolo[3,4-b]pyrrole-5(1H)-carbonitrile

[0322] ##STR00045##

[0323] To a stirred solution rac-(3aR,6aR)-1-((4-(pyridin-3-yl)phenyl)sulfonyl) octahydropyrrolo[3,4-b]pyrrole TFA salt (0.200 g, 0.45 mmol) in THF (20 mL) was added K.sub.2CO.sub.3 (0.333 g, 2.41 mmol) at 0 C. followed by the addition of CNBr (0.051 g, 0.48 mmol) at the same temperature. The reaction mixture was stirred at room temperature for 1 h. The resulting reaction mixture was concentrated under reduced pressure and resulting residue was purified by flash column chromatography (eluting with 3.0% MeOH in DCM) yielding the title compound (0.150 g, 0.423 mmol). LCMS: Method A, 3.698 min, MS: ES+354.9 [M+1]; .sup.1H NMR (400 MHz, DMSO-D6): 8.96 (d, J=2 Hz, 1H), 8.65-8.64 (m, 1H), 8.19-8.17 (m, 1H), 8.02 (d, J, 8.4 Hz, 2H), 7.95 (d, J=8.8 Hz, 2H), 7.54-7.57 (m, 1H), 4.05-4.01 (m, 1H), 3.58 (d, J=7.2 Hz, 2H), 3.56-3.48 (m, 1H), 3.41-3.28 (m, 1H), 3.35 (dd, J=10.0 & 6.0 Hz, 1H), 3.18-3.14 (m, 1H), 1.74-1.62 (m, 1H), 1.65-1.71 (m, 2H).

EXAMPLE 22

rac-(4aR,7aR)-4-tosylhexahydropyrrolo[3,4-b][1,4]oxazine-6(2H)-carbonitrile

[0324] ##STR00046##

[0325] The title compound was synthesised via general method A using pyrrolo[3,4-b]-1,4-oxazine-6(2H)-carboxylic acid, hexahydro-1,1-dimethylethyl ester, (4aR,7aS) in step a. LCMS: Method D, 2.591 min, MS: ES+345.1 [M+1].

EXAMPLE 23

rac-(4aR,7aS)-4-(4-methylbenzyl)sulfonyl)hexahydropyrrolo[3,4-b][1,4]oxazine-6(2H)-carbonitrile

[0326] ##STR00047##

[0327] The title compound was synthesised via general method A using pyrrolo[3,4-b]-1,4-oxazine-6(2H)-carboxylic acid, hexahydro-1,1-dimethylethyl ester, (4aR,7aS) in step a. LCMS: Method D, 2.591 min, MS: ES+322.4 [M+1].

EXAMPLE 24

N-(5-(N-(1-cyanopyrrolidin-3-yl)sulfamoyl)pyridin-2-yl)-1-methyl-1H-pyrazole-5-carboxamide

[0328] ##STR00048##

[0329] Step 1. pyridin-2-amine was heated to 150 C. in chlorosulfonic acid to afford 6-aminopyridine-3-sulfonyl chloride which was used crude in the next reaction.

[0330] Step 2. 6-aminopyridine-3-sulfonyl chloride was added to tert-butyl 3-aminopyrrolidine-1-carboxylate using general method A, step a to afford t-butyl 3-((6-aminopyridine)-3-sulfonamido)pyrrolidine-1-carboxylate.

[0331] Step 3. t-butyl 3-((6-aminopyridine)-3-sulfonamido)pyrrolidine-1-carboxylate and 1-methyl-1H-pyrazole-5-carboxylic acid were heated to 70 C. with 50% T3P in ethyl acetate, triethylamine and THF for 30 h to afford tert-butyl 3-((6-(1-methyl-1H-pyrazole-5-carboxamido)pyridine)-3-sulfonamido)pyrrolidine-1-carboxylate.

[0332] Step 4. The title compound was synthesised following the procedure of general method A, steps b-c. LCMS: Method B, 2.998 min, MS: ES+376.13 [M+1]; .sup.1H NMR (400 MHz, DMSO-d6) 11.32 (s, 1H), 8.77 (d, J=2.4 Hz, 1H), 8.37 (d, J=8.4 Hz, 1H), 8.24-8.26 (m, 2H), 7.55 (d, J=2 Hz, 1H), 7.34 (d, J=2 Hz, 1H), 4.1 (s, 3H), 3.78-3.82 (m, 1H), 3.31-3.46 (m, 3H), 3.09-3.12 (m, 1H), 1.88-1.99 (m, 1H), 1.66-1.74 (m, 1H).

EXAMPLE 25

N-(1-cyanopyrrolidin-3-yl)-N-methyl-4-((5-(trifluoromethyl)pyridin-2-yl)oxy)benzenesulfonamide

[0333] ##STR00049##

[0334] The title compound was synthesised via general method C using tert-butyl 3-(methylamino)pyrrolidine-1-carboxylate in step a. LCMS: Method D, 3.046 min, MS: ES+427.2 [M+1].

Compounds in Table 4 were synthesised according to General Method D.

##STR00050##

TABLE-US-00010 TABLE 4 LCMS LCMS RT MS Ex RRN Name Method (min) (ES+) 26 [00051] N-([1,1-biphenyl]-4-yl)-1-cyanopyrrolidine- 3-sulfonamide B 4.193 326.2 (ES) 27 [00052] 3-((4-(4-fluorophenyl)piperazin-1- yl)sulfonyl)pyrrolidine-1-carbonitrile D 2.603 339.1 28 [00053] (3-((4-(pyridin-2-yl)piperazin-1- yl)sulfonyl)pyrrolidine-1-carbonitrile E 2.162 322.1 29 [00054] 3-((4-(pyrimidin-2-yl)piperazin-1- yl)sulfonyl)pyrrolidine-1-carbonitrile E 1.889 323.1 30 [00055] 3-((4-(4-chlorophenyl)piperidin-1- yl)sulfonyl)pyrrolidine-1-carbonitrile D 2.824 354.1 31 [00056] (S)-3-((4-(4-chlorophenyl)piperidin-1- yl)sulfonyl)pyrrolidine-1-carbonitrile B 4.586 370.5 32 [00057] (R)-3-((4-(4-chlorophenyl)piperidin-1- yl)sulfonyl)pyrrolidine-1-carbonitrile B 4.561 370.5 33 [00058] 3-((4-benzylpiperidin-1- yl)sulfonyl)pyrrolidine-1-carbonitrile D 3.107 334.2 34 [00059] 3-((3-(4-chlorophenyl)azetidin-1- yl)sulfonyl)pyrrolidine-1-carbonitrile B 4.129 326.3 35 [00060] (S)-3-((3-(4-chlorophenyl)azetidin-1- yl)sulfonyl)pyrrolidine-1-carbonitrile B 4.211 326.3 36 [00061] (R)-3-((3-(4-chlorophenyl)azetidin-1- yl)sulfonyl)pyrrolidine-1-carbonitrile B 4.220 326.3 37 [00062] 3-((3-phenoxyazetidin-1- yl)sulfonyl)pyrrolidine-1-carbonitrile A 4.239 308.0

Biological Activity of Compounds of the Invention

Abbreviations

[0335] TAMRA carboxytetramethylrhodamine [0336] PCR polymerase chain reaction [0337] PBS phosphate buffered saline [0338] EDTA ethylenediaminetetraacetic acid [0339] Tris 2-amino-2-(hydroxymethyl)-1,3-propanediol [0340] NP-40 Nonidet P-40, octylphenoxypolyethoxyethanol [0341] BSA bovine serum albumin [0342] PNS peripheral nervous system [0343] BH3 Bcl-2 homology domain 3 [0344] PTEN phosphatase and tensin homologue

In Vitro UCHL1 Inhibition Assay

Expression and Purification of UCHL1

[0345] The UCHL1 construct was PCR amplified and cloned into a pFLAG-CMV-6a vector (Sigma-Aldrich) with an N-terminal FLAG tag. HEK293T cells were transfected with FLAG-UCHL1 using TransIT-LT1 transfection reagent (Mirus) according to the manufacturer's instructions. Cells were harvested 40 hours after transfection. Cells were washed once with PBS and scraped in lysis buffer (50 mM Tris, pH 7.5, 150 mM NaCl, 3 mM EDTA, 0.5% NP40, 10% glycerol, 5 mM beta-mercaptoethanol, protease inhibitors (complete mini, Roche) and phosphatase inhibitors (PhosSTOP mini, Roche). Lysates were incubated for 30 min on ice and centrifuged at 1200 rpm for 10 min at 4 C. Soluble supernatant was added to FLAG affinity resin (EZview Rad ANTI-FLAG M2 affinity gel, Sigma-Aldrich) equilibrated in low salt buffer (20 mM Tris, pH 7.5, 150 mM NaCl, 0.5 mM EDTA, 5 mM beta-mercaptoethanol) and incubated at 4 C. for 3 hours rotating. The resin was spun at 2000 rpm for 2 min and the supernatant was removed. The resin was washed two times with low salt buffer and one time with high salt buffer (20 mM Tris, pH 7.5, 500 mM NaCl, 0.5 mM EDTA, 5 mM beta-mercaptoethanol, protease inhibitors (complete mini, Roche) and phosphatase inhibitors (PhosSTOP mini, Roche). To elute the bound UCHL1, elution buffer (10 mM Tris, pH 7.5, 150 mM NaCl, 0.5 mM EDTA, 10% glycerol, 0.5% NP40, 5 mM beta-mercaptoethanol, 0.15 mg/ml 3 FLAG peptide (Sigma-Aldrich)) was added to the resin and incubated at 4 C. for 2.5 hours rotating. The resin was centrifuged at 4000 rpm for 30 seconds, and the supernatant containing purified FLAG-UCHL1 was removed and stored at 80 C.

UCHL1 Biochemical Kinetic Assay

[0346] Reactions were performed in duplicate in black 384 well plates (small volume, Greiner 784076) in a final reaction volume of 21l. UCHL1 was diluted in reaction buffer (20 mM Tris, pH 7.5, 100 mM NaCl, 0.05% Tween 20, 0.5 mg/ml BSA, 5 mM beta-mercaptoethanol) to the equivalent of 0, 0.01, 0.05, 0.1, 0.5, and 1 l/well. Buffer was optimised for optimal temperature, pH, reducing agent, salts, time of incubation, and detergent. Reactions were initiated by the addition of 50 nM of TAMRA labelled peptide linked to ubiquitin via an iso-peptide bond as fluorescence polarisation substrate. Reactions were incubated at room temperature and read every 2 min for 120 min. Readings were performed on a Pherastar Plus (BMG Labtech). Excitation 540 nm; Emission 590 nm.

UCHL1 Biochemical IC50 Assay

[0347] Dilution plates were prepared at 21 times the final concentration (2100 M for a final concentration of 100 M) in 50% DMSO in a 96-well polypropylene V-bottom plate (Greiner #651201). A typical 8-point dilution series to be 30, 10, 3, 1, 0.3, 0.1, 0.03, 0.01 M final. Reactions were performed in duplicate in black 384 well plates (small volume, Greiner 784076) in a final reaction volume of 21 l. Either 1 l of 50% DMSO or diluted compound was added to the plate. UCHL1 was diluted in reaction buffer (20 mM Tris, pH 7.5, 100 mM NaCl, 0.05% Tween 20, 0.5 mg/ml BSA, 5 mM beta-mercaptoethanol) to the equivalent of 0.05 l/well and 10 l of diluted UCHL1 was added to the compound. Enzyme and compound were incubated for 30 min at room temp. Reactions were initiated by the addition of 50 nM of TAMRA labelled peptide linked to ubiquitin via an iso-peptide bond as fluorescence polarisation substrate. Reactions were read immediately after addition of substrate and following a 2-hour incubation at room temperature. Readings were performed on a Pherastar Plus (BMG Labtech). Excitation 540 nm; Emission 590 nm.

Activity of Exemplary Compounds in UCHL1 Biochemical 1050 Assay

Ranges:

[0348] A<0.1 M; [0349] 0.1<B<1 M; [0350] 1<C<10 M; [0351] 10 M<D<100 M

TABLE-US-00011 Example IC50 range 1 D 2 C 3 D 4 C 5 D 6 D 8 C 9 C 11 D 12 C 13 C 14 D 15 C 16 D 18 D 19 B 20 C 24 D 25 C 26 D

In Vitro USP30 Inhibition Assay

[0352] USP30 biochemical kinetic assay. Reactions were performed in duplicate in black 384 well plates (small volume, Greiner 784076) in a final reaction volume of 21l. USP30 CD (57-517, #64-0057-050 Ubiquigent) was diluted in reaction buffer (40 mM Tris, pH 7.5, 0.005% Tween 20, 0.5 mg/ml BSA, 5 mM beta-mercaptoethanol) to the equivalent of 0, 0.005, 0.01, 0.05, 0.1 and 0.5 l/well. Buffer was optimised for optimal temperature, pH, reducing agent, salts, time of incubation, and detergent. Reactions were initiated by the addition of 50 nM of TAMRA labelled peptide linked to ubiquitin via an iso-peptide bond as fluorescence polarisation substrate. Reactions were incubated at room temperature and read every 2 min for 120 min. Readings were performed on a Pherastar Plus (BMG Labtech). Excitation 540 nm; Emission 590 nm.

USP30 Biochemical 1050 Assay

[0353] Dilution plates were prepared at 21 times the final concentration (2100 M for a final concentration of 100 M) in 50% DMSO in a 96-well polypropylene V-bottom plate (Greiner #651201). A typical 8-point dilution series would be 100, 30, 10, 3, 1, 0.3, 0.1, 0.03 M final. Reactions were performed in duplicate in black 384 well plates (small volume, Greiner 784076) in a final reaction volume of 21 pI. Either 1 l of 50% DMSO or diluted compound was added to the plate. USP30 was diluted in reaction buffer (40 mM Tris, pH 7.5, 0.005% Tween 20, 0.5 mg/ml BSA, 5 mM beta-mercaptoethanol) to the equivalent of 0.05 l/well and 10 l of diluted USP30 was added to the compound. Enzyme and compound were incubated for 30 min at room temp. Reactions were initiated by the addition of 50 nM of TAMRA labelled peptide linked to ubiquitin via an iso-peptide bond as fluorescence polarisation substrate. Reactions were read immediately after addition of substrate and following a 2-hour incubation at room temperature. Readings were performed on a Pherastar Plus (BMG Labtech). Excitation 540 nm; Emission 590 nm.

Activity of Exemplary Compounds in USP30 Biochemical IC50 Assay

Ranges:

[0354] A<0.1 M; [0355] 0.1<B<1 M; [0356] 1<C<10 M; [0357] 10 M<D<100 M

TABLE-US-00012 Example IC50 range 1 B 2 B 3 B 4 B 6 C 7 C 8 B 9 B 10 C 11 B 13 B 14 B 15 B 16 B 17 C 18 C 19 B 20 B 21 B 22 D 23 B 24 B 25 B 26 B 27 D 28 B 29 C 30 A 31 A 32 A 33 B 34 B 35 B 36 B 37 D