DERIVATIVES OF ITACONIC ACID AND THEIR USE AS ANTI-INFLAMMATORY AGENTS

Abstract

The invention relates to compounds of formula (I) and to their use in treating or preventing an inflammatory disease or a disease associated with an undesirable immune response:

##STR00001## wherein R.sup.A and R.sup.B are as defined herein.

Claims

1: A compound of formula (I): ##STR00060## wherein the moiety: ##STR00061## represents ##STR00062## R.sup.A is: ##STR00063## wherein: R.sup.A1 is selected from the group consisting of H, halo, C.sub.1-4 alkyl, C.sub.1-4 haloalkyl, C.sub.1-4 alkoxy, C.sub.1-4 haloalkoxy, SC.sub.1-4 alkyl, SC.sub.1-4 haloalkyl and SF.sub.5; R.sup.A2 is selected from the group consisting of halo, C.sub.1-4 alkyl, C.sub.1-4 haloalkyl, C.sub.1-4 alkoxy and C.sub.1-4 haloalkoxy; R.sup.A3 is C.sub.1-2 alkyl; m is 0, 1 or 2; n is 1 or 2; and p is 0, 1, 2, 3 or 4; or R.sup.A is: ##STR00064## wherein: R.sup.A4 is H, C.sub.1-4 alkyl, C.sub.1-4haloalkyl or CCH; and R.sup.A5 is H, C.sub.1-4 alkyl or C.sub.1-4 haloalkyl; or R.sup.A4 and R.sup.A5 join to form a C.sub.3-6 cycloalkyl ring or a 4-6 membered heterocyclic ring wherein the C.sub.3-6 cycloalkyl ring is optionally substituted by one or more R.sup.A6, wherein R.sup.A6 is independently selected from the group consisting of fluoro, methyl and cyano, or two R.sup.A6 groups which are attached to the same carbon atom join to form a C.sub.3-4 cycloalkyl ring; A is phenyl or 5-6-membered heteroaryl wherein when A is phenyl or 6-membered heteroaryl A is optionally substituted by one or more R.sup.A7, and when A is 5-membered heteroaryl A is substituted by one or more R.sup.A7, wherein when A is phenyl substituted by one or more R.sup.A7, one R.sup.A7 is in the 4-position with respect to C(R.sup.A4)(R.sup.A5); R.sup.A7 is independently selected from the group consisting of halo, C.sub.1-4 alkyl, C.sub.1-4 haloalkyl, C.sub.1-4 haloalkoxy, SF.sub.5, SC.sub.1-4 alkyl and SC.sub.1-4 haloalkyl; and R.sup.B is NR.sup.B1R.sup.B2 or OR.sup.B3; wherein: R.sup.B1 and R.sup.B2 are independently selected from the group consisting of H, C.sub.1-4 alkyl and C.sub.1-4 haloalkyl and 4-7-membered heterocyclyl wherein R.sup.B1 and/or R.sup.B2 are optionally substituted by one or more R.sup.B4 wherein R.sup.B4 is selected from the group consisting of C.sub.1-2 alkyl, NH.sub.2, N(C.sub.1-2 alkyl).sub.2, hydroxy, oxo, 5-7-membered heterocyclyl and 5-6-membered heteroaryl optionally substituted by C.sub.1-2 alkyl; and R.sup.B3 is selected from the group consisting of H, C.sub.1-4 alkyl, C.sub.1-4 haloalkyl, CH.sub.2COOH, CH.sub.2CH.sub.2COOH, CH.sub.2tetrazolyl and CH.sub.2CH.sub.2tetrazolyl, wherein R.sup.B3 is optionally substituted on an available carbon atom by one or more R.sup.B3 wherein R.sup.B3 is selected from the group consisting of difluoromethyl, trifluoromethyl and methyl; and/or wherein R.sup.B3 is optionally substituted by two R.sup.B3 groups which are attached to the same carbon atom and join to form a C.sub.3-6 cycloalkyl or a 4-6-membered heterocyclyl ring; or a pharmaceutically acceptable salt and/or solvate thereof.

2: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 1 wherein the moiety: ##STR00065## represents ##STR00066##

3. (canceled)

4. The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 1 wherein R.sup.A is: ##STR00067## wherein R.sup.A1, R.sup.A2, R.sup.A3, m, n and p are defined in claim 1.

5: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 4 wherein R.sup.A1 is C.sub.1-4 haloalkyl.

6: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 4 wherein m is 0.

7: The compound, pharmaceutically acceptable salt and/or solvate thereof according claim 4 wherein n is 1.

8: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 4 wherein p is 0.

9: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 1 wherein R.sup.A is: ##STR00068## wherein A, R.sup.A4 and R.sup.A5 are defined in claim 1.

10: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 9 wherein R.sup.A4 is C.sub.1-4 alkyl such as methyl.

11: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 9 wherein R.sup.A5 is H.

12: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 9 wherein when R.sup.A4 and R.sup.A5 are different, the carbon atom to which R.sup.A4 and R.sup.A5 are attached has the following stereochemistry: ##STR00069##

13: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 9 wherein R.sup.A4 and R.sup.A5 join to form a C.sub.3-6 cycloalkyl ring or a 4-6 membered heterocyclic.

14: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 13 wherein the C.sub.3-6 cycloalkyl ring is not substituted; or wherein the C.sub.3-6 cycloalkyl ring is substituted by one or more R.sup.A6, wherein R.sup.A6 is independently selected from the group consisting of fluoro, methyl and cyano, or two R.sup.A6 groups which are attached to the same carbon atom join to form a C.sub.3-4 cycloalkyl ring.

15-16. (canceled)

17: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 9 wherein A is phenyl, thienyl or pyridinyl.

18. (canceled)

19: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 9 wherein A is unsubstituted phenyl or unsubstituted 6-membered heteroaryl.

20: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 9 wherein A is substituted by one or more, wherein R.sup.A7 is defined in claim 1, and wherein when A is phenyl substituted by one or more R.sup.A7, one R.sup.A7 is in the 4-position with respect to C(R.sup.A4)(R.sup.A5).

21: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 20 wherein R.sup.A7 is halo or C.sub.1-4 haloalkyl.

22. (canceled)

23: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 1 wherein R.sup.B is NR.sup.B1R.sup.B2.

24-36. (canceled)

37: The compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 1 wherein R.sup.B is OR.sup.B3.

38-45. (canceled)

46: The compound according to claim 1, which is selected from the list consisting of: (S)-3-((1-(4-(trifluoromethyl)phenyl)ethoxy)carbonyl)but-3-enoic acid; 3-((1-(4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoic acid; 3-((1-(3-fluoro-4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoic acid; 3-(((3-(4-(trifluoromethyl)phenyl)oxetane-3-yl)oxy)carbonyl)but-3-enoic acid; 3-((1-(5-(trifluoromethyl)pyridin-2-yl)cyclobutoxy)carbonyl)but-3-enoic acid; 3-((1-(5-(trifluoromethyl)thiophen-2-yl)cyclobutoxy)carbonyl)but-3-enoic acid; 3-(((6-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-yl)oxy)carbonyl)but-3-enoic acid; 4-methyl 1-(1-(4-(trifluoromethyl)phenyl)cyclobutyl) 2-methylenesuccinate; 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(methylamino)-2-methylene-4-oxobutanoate; 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(dimethylamino)-2-methylene-4-oxobutanoate; 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-amino-2-methylene-4-oxobutanoate; 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((2-(dimethylamino)ethyl)amino)-2-methylene-4-oxobutanoate; 1-(4-(trifluoromethyl)phenyl)cyclobutyl 2-methylene-4-((2-morpholinoethyl)amino)-4-oxobutanoate; 1-(4-(trifluoromethyl)phenyl)cyclobutyl 2-methylene-4-((1-methylpiperidin-4-yl)amino)-4-oxobutanoate; 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(((1H-imidazol-2-yl)methyl)amino)-2-methylene-4-oxobutanoate; 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(((1-methyl-1H-imidazol-2-yl)methyl)amino)-2-methylene-4-oxobutanoate; 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((1-amino-3-hydroxy-1-oxopropan-2-yl)amino)-2-methylene-4-oxobutanoate; 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((1,1-dioxidothietan-3-yl)amino)-2-methylene-4-oxobutanoate; 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((2-hydroxyethyl)amino)-2-methylene-4-oxobutanoate; 1-(4-(trifluoromethyl)phenyl)cyclobutyl 2-methylene-4-oxo-4-((tetrahydro-2H-pyran-4-yl)amino)butanoate; 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((2-amino-2-oxoethyl)amino)-2-methylene-4-oxobutanoate; 2-((3-((1-(4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoyl)oxy)acetic acid; and 3-methyl-4-oxo-4-(1-(4-(trifluoromethyl)phenyl)cyclobutoxy)but-2-enoic acid; or a pharmaceutically acceptable salt and/or solvate of any one thereof.

47: A pharmaceutical composition comprising a compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 1.

48-50. (canceled)

51: A method of treating or preventing an inflammatory disease or a disease associated with an undesirable immune response, which comprises administering a compound, pharmaceutically acceptable salt and/or solvate thereof according to claim 1.

52-56. (canceled)

57: The method according to claim 51, wherein the inflammatory disease or disease associated with an undesirable immune response is selected from the group consisting of rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, systemic lupus erythematosus, multiple sclerosis, psoriasis, Crohn's disease, ulcerative colitis, uveitis, cryopyrin-associated periodic syndromes, Muckle-Wells syndrome, juvenile idiopathic arthritis, chronic obstructive pulmonary disease and asthma.

58-68. (canceled)

Description

DETAILED DESCRIPTION OF THE INVENTION

Compounds of Formula (I)

[0034] Embodiments and preferences set out herein with respect to the compound of formula (I) apply equally to the pharmaceutical composition, compound for use, use and method aspects of the invention.

[0035] As used herein, the term C.sub.1-4 alkyl refers to a straight or branched fully saturated hydrocarbon group having the specified number of carbon atoms. The term encompasses methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl and tert-butyl. The term alkyl also encompasses alkylene which is a bifunctional straight or branched fully saturated hydrocarbon group having the stated number of carbon atoms. Example alkylene groups include methylene, ethylene, n-propylene and n-butylene.

[0036] The term C.sub.1-4 alkoxy refers to an alkyl group, such as those defined above, singularly bonded via an oxygen atom. Examples of alkoxy groups include OCH.sub.3.

[0037] The term halo refers to fluorine, chlorine, bromine or iodine. Particular examples of halo are fluorine, chlorine and bromine, especially fluorine.

[0038] The term C.sub.1-4 haloalkyl refers to a straight or a branched fully saturated hydrocarbon chain containing the specified number of carbon atoms and at least one halogen atom, such as fluoro or chloro, especially fluoro. An example of haloalkyl is CF.sub.3. Further examples of haloalkyl are CHF.sub.2, CF.sub.2CH.sub.3 and CH.sub.2CF.sub.3.

[0039] The term C.sub.1-4 haloalkoxy refers to a haloalkyl group as defined above, singularly bonded via an oxygen atom. Examples of haloalkoxy groups include OCF.sub.3, OCHF.sub.2 and OCH.sub.2CF.sub.3.

[0040] The term C.sub.3-6 cycloalkyl such as C.sub.3-4 cycloalkyl refers to a fully saturated cyclic hydrocarbon group having the specified number of carbon atoms. The term encompasses cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0041] The term 4-7-membered heterocyclic ring (such as 4-6-membered heterocyclic ring or 5-7-membered heterocyclic ring) refers to a non-aromatic cyclic group having the stated number of ring atoms and wherein at least one of the ring atoms is a heteroatom selected from N, O, S and B. The term heterocyclic ring is interchangeable with heterocyclyl. The term encompasses azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, oxepanyl, thiepanyl and azepanyl. 4-7-membered heterocyclyl groups can typically be substituted by one or more (e.g. one or two) oxo groups. Suitably, a sulphur atom is substituted by one or two oxo groups thus forming SO or SO.sub.2.

[0042] The term 5-6-membered heteroaryl refers to a cyclic group with aromatic character wherein at least one of the atoms in the cyclic group is a heteroatom independently selected from N, O and S. The term encompasses pyrrolyl, furanyl, thienyl, imidazolyl, pyrazolyl, thiazolyl, oxadiazolyl, thiadiazolyl, triazolyl, oxazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyradizinyl and pyrazinyl.

[0043] The term tetrazolyl refers to a 5-(1H-tetrazolyl) substituent where the tetrazole is linked to the rest of the molecule via a carbon atom:

##STR00007##

wherein the dashed line indicates the point of attachment to the rest of the molecule.

[0044] Where substituents are indicated as being optionally substituted in formula (I) in the embodiments and preferences set out below, the optional substituent may be attached to an available carbon atom, which means a carbon atom which is attached to a hydrogen atom i.e. a CH group. The optional substituent replaces the hydrogen atom attached to the carbon atom.

[0045] In one embodiment, the moiety:

##STR00008##

represents

##STR00009##

[0046] The carbon-carbon double bond in this structure is referred to as exo.

[0047] In another embodiment, the moiety:

##STR00010##

represents

##STR00011##

[0048] The carbon-carbon double bond in this structure is referred to as endo.

[0049] In the endo embodiment, the double bond may be cis or trans such that both of the following moieties are covered:

##STR00012##

[0050] Suitably, the endo double bond in the compound of formula (I) is trans.

[0051] The compounds of formula (I) in which the carbon-carbon double bond is endo can generally be obtained by isomerisation from compounds of formula (I) in which the carbon-carbon double bond is exo and such isomerisation may occur in in vitro assays or in vivo following administration of the exo compound. In some cases, isomerisation in in vitro assays, such as in vitro hepatocyte stability assays, or in vivo following administration of the exo compound may be partial and thus lead to a mixture of the endo and exo compounds resulting. In some cases, the mixture of endo and exo isomers may contribute to the activity observed in a particular assay. Suitably, compounds of formula (I), such as those in which the carbon-carbon double bond is exo, are stable to isomerisation.

[0052] In one embodiment, R.sup.A is:

##STR00013##

wherein R.sup.A1, R.sup.A2, R.sup.A3, m, n and p are defined above.

[0053] In one embodiment, R.sup.A1 is C.sub.1-4 haloalkyl such as CF.sub.3.

[0054] In one embodiment, m is 0.

[0055] In one embodiment, n is 1.

[0056] In one embodiment, p is 0.

[0057] In another embodiment, R.sup.A is:

##STR00014##

wherein A, R.sup.A4 and R.sup.A5 are defined above.

[0058] In one embodiment, R.sup.A4 is C.sub.1-4 alkyl such as methyl.

[0059] In one embodiment, R.sup.A5 is H.

[0060] Suitably, when R.sup.A4 and R.sup.A5 are different (e.g. when R.sup.A4 is H and R.sup.A5 is methyl), the carbon atom to which R.sup.A4 and R.sup.A5 are attached has the following stereochemistry:

##STR00015##

[0061] In one embodiment, R.sup.A4 and R.sup.A5 join to form a C.sub.3-6 cycloalkyl ring such as a C.sub.4 cycloalkyl ring.

[0062] In one embodiment, the C.sub.3-6 cycloalkyl ring is not substituted. In one embodiment, the C.sub.3-6 cycloalkyl ring is substituted by one or more (such as one, two or three e.g. one) R.sup.A6, wherein R.sup.A6 is independently selected from the group consisting of fluoro, methyl and cyano, or two R.sup.A6 groups which are attached to the same carbon atom join to form a C.sub.3-4 cycloalkyl ring.

[0063] In one embodiment, R.sup.A4 and R.sup.A5 join to form a 4-6 membered heterocyclic ring such as a 4-membered heterocyclic ring e.g., oxetanyl.

[0064] In one embodiment, A is phenyl. In a second embodiment, A is 5-6-membered heteroaryl, such as thienyl or pyridinyl. Suitably, A is 5-membered heteroaryl. Alternatively, A is 6-membered heteroaryl.

[0065] In one embodiment, and when A is phenyl or 6-membered heteroaryl, A is not substituted. In another embodiment, A is phenyl or 5-6-membered heteroaryl and is substituted by one or more (such as one, two or three e.g. one) R.sup.A7, wherein R.sup.A7 is defined above.

[0066] In one embodiment, R.sup.A7 is halo such as F. In a second embodiment, R.sup.A7 is C.sub.1-4 haloalkyl such as CF.sub.3.

[0067] When A is phenyl substituted with one or more R.sup.A7, one R.sup.A7 is in the 4-position with respect to C(R.sup.A4)(R.sup.A5):

##STR00016##

[0068] In this embodiment, suitably, R.sup.A7 is C.sub.1-4 haloalkyl such as CF.sub.3.

[0069] In one embodiment, R.sup.B is NR.sup.B1R.sup.B2. In a second embodiment, R.sup.B is OR.sup.B3.

[0070] In one embodiment, R.sup.B1 is H. In a second embodiment, R.sup.B1 is C.sub.1-4 alkyl such as methyl. In a third embodiment, R.sup.B1 is C.sub.1-4 haloalkyl. In a fourth embodiment, R.sup.B1 is 4-7-membered heterocyclyl such as tetrahydropyranyl or optionally substituted thietanyl, for example dioxidothietanyl. In some cases, R.sup.B1 is 5-7-membered heterocyclyl such as tetrahydropyranyl.

[0071] In one embodiment, R.sup.B2 is H. In a second embodiment, R.sup.B2 is C.sub.1-4 alkyl such as methyl. In a third embodiment, R.sup.B2 is C.sub.1-4 haloalkyl. In a fourth embodiment, R.sup.B2 is 4-7-membered heterocyclyl such as tetrahydropyranyl or optionally substituted thietanyl, for example dioxidothietanyl. In some cases, R.sup.B2 is 5-7-membered heterocyclyl such as tetrahydropyranyl.

[0072] Suitably, both R.sup.B1 and R.sup.B2 are H, or both R.sup.B1 and R.sup.B2 are methyl. Alternatively, R.sup.B1 is H and R.sup.B2 is methyl.

[0073] Suitably, at least one of R.sup.B1 and R.sup.B2 is H.

[0074] In one embodiment, R.sup.B1 and/or R.sup.B2 are not substituted. In another embodiment, R.sup.B1 and/or R.sup.B2 are substituted by one or more (such as one, two or three, e.g., one) R.sup.B4 wherein R.sup.B4 is selected from the group consisting of C.sub.1-2 alkyl, NH.sub.2, N(C.sub.1-2 alkyl).sub.2, hydroxy, oxo, 5-7-membered heterocyclyl and 5-6-membered heteroaryl optionally substituted by C.sub.1-2 alkyl.

[0075] In one embodiment, R.sup.B4 is C.sub.1-2 alkyl. In a second embodiment, R.sup.B4 is NH.sub.2. In a third embodiment, R.sup.B4 is N(C.sub.1-2 alkyl).sub.2. In a fourth embodiment, R.sup.B4 is hydroxy. In a fifth embodiment, R.sup.B4 is oxo (O). In a sixth embodiment, R.sup.B4 is 5-7-membered heterocyclyl. In a seventh embodiment, R.sup.B4 is 5-6-membered heteroaryl optionally substituted by C.sub.1-2 alkyl.

[0076] In one embodiment R.sup.B is OR.sup.B3.

[0077] In one embodiment, R.sup.B3 is H. In a second embodiment, R.sup.B3 is C.sub.1-4 alkyl such as methyl. In a third embodiment, R.sup.B3 is CH.sub.2COOH.

[0078] In one embodiment, R.sup.B3 is not substituted. In another embodiment, R.sup.B3 is optionally substituted on an available carbon atom by one or more (such as one, two or three e.g. one) R.sup.B3 wherein R.sup.B3 is selected from the group consisting of difluoromethyl, trifluoromethyl and methyl; and/or wherein R.sup.B3 is optionally substituted by two R.sup.B3 groups which are attached to the same carbon atom and join to form a C.sub.3-6 cycloalkyl or a 4-6-membered heterocyclyl ring.

[0079] The term available carbon atom means any carbon atom which forms a CH bond. The substituent replaces the hydrogen atom attached to the carbon atom. The skilled person will appreciate that when R.sup.B3 is H, there is no available carbon atom for substitution.

[0080] In one embodiment, the molecular weight of the compound of formula (I) is 150 Da-450 Da, suitably 200 Da-400 Da.

[0081] In one embodiment there is provided a compound of formula (I), selected from the group consisting of: [0082] (S)-3-((1-(4-(trifluoromethyl)phenyl)ethoxy)carbonyl)but-3-enoic acid; [0083] 3-((1-(4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoic acid; [0084] 3-((1-(3-fluoro-4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoic acid; [0085] 3-(((3-(4-(trifluoromethyl)phenyl)oxetane-3-yl)oxy)carbonyl)but-3-enoic acid; [0086] 3-((1-(5-(trifluoromethyl)pyridin-2-yl)cyclobutoxy)carbonyl)but-3-enoic acid; [0087] 3-((1-(5-(trifluoromethyl)thiophen-2-yl)cyclobutoxy)carbonyl)but-3-enoic acid; [0088] 3-(((6-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-yl)oxy)carbonyl)but-3-enoic acid; [0089] 4-methyl 1-(1-(4-(trifluoromethyl)phenyl)cyclobutyl) 2-methylenesuccinate; [0090] 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(methylamino)-2-methylene-4-oxobutanoate; [0091] 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(dimethylamino)-2-methylene-4-oxobutanoate; [0092] 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-amino-2-methylene-4-oxobutanoate; [0093] 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((2-(dimethylamino)ethyl)amino)-2-methylene-4-oxobutanoate; [0094] 1-(4-(trifluoromethyl)phenyl)cyclobutyl 2-methylene-4-((2-morpholinoethyl)amino)-4-oxobutanoate; [0095] 1-(4-(trifluoromethyl)phenyl)cyclobutyl 2-methylene-4-((1-methylpiperidin-4-yl)amino)-4-oxobutanoate; [0096] 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(((1H-imidazol-2-yl)methyl)amino)-2-methylene-4-oxobutanoate; [0097] 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(((1-methyl-1H-imidazol-2-yl)methyl)amino)-2-methylene-4-oxobutanoate; [0098] 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((1-amino-3-hydroxy-1-oxopropan-2-yl)amino)-2-methylene-4-oxobutanoate; [0099] 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((1,1-dioxidothietan-3-yl)amino)-2-methylene-4-oxobutanoate; [0100] 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((2-hydroxyethyl)amino)-2-methylene-4-oxobutanoate; [0101] 1-(4-(trifluoromethyl)phenyl)cyclobutyl 2-methylene-4-oxo-4-((tetrahydro-2H-pyran-4-yl)amino)butanoate; [0102] 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((2-amino-2-oxoethyl)amino)-2-methylene-4-oxobutanoate; [0103] 2-((3-((1-(4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoyl)oxy)acetic acid; and [0104] 3-methyl-4-oxo-4-(1-(4-(trifluoromethyl)phenyl)cyclobutoxy)but-2-enoic acid;
or a pharmaceutically acceptable salt and/or solvate of any one thereof.

[0105] Compounds of formula (I) may be prepared as set out in the Examples and as set out in the following schemes.

##STR00017##

wherein R.sup.A, R.sup.B1, R.sup.B2 and R.sup.B3 are defined elsewhere herein, P is a carboxylic acid protecting group such as CH.sub.2CCl.sub.3, and X is a leaving group such as halo, e.g., bromo.

[0106] Step 1: Lewis acid catalysed ring opening of itaconic anhydride (VI) using HOP provides ,-unsaturated carboxylic acids (V).

[0107] Step 2: Esterification of carboxylic acids (V) with alcohols (IV) under standard conditions (such as DCC, DMAP in DCM) provides ,-unsaturated esters of formula (III). Alcohols (IV) are commercially available or can be made according to the Examples.

[0108] Step 3: Protecting group P is removed under conditions known to the person skilled in the art to give compounds of formula (I) wherein R.sup.B3 is H. For example, when P is CH.sub.2CCl.sub.3, Zn/NH.sub.4OAc may be used to remove this protecting group.

[0109] Step 4: Carboxylic acids of formula (I), wherein R.sup.B3 is H, may be converted to compounds of formula (I), wherein R.sup.B is NR.sup.B1R.sup.B2, using standard amide coupling conditions, such as using T3P, a base, such as Et.sub.3N, in a solvent, such as EtOAc, or using HATU in the presence of a base, such as TEA, in a solvent such as dimethylformamide.

[0110] Step 5: Compounds of formula (II) may be obtained by reacting compounds of formula (I), wherein R.sup.B3 is H, with a protected carboxylic acid derivative (VII).

[0111] Step 6: Removal of protecting group P under conditions known to the person skilled in the art provides compounds of formula (I) wherein R.sup.B3 is CH.sub.2COOH. For example, when P is CH.sub.2CCl.sub.3, Zn/NH.sub.4OAc may be used to remove this protecting group.

[0112] Step 7: Carboxylic acids of formula (I), wherein R.sup.B3 is H, may be converted to compounds of formula (I), wherein R.sup.B is OR.sup.B3, by esterification of carboxylic acids of formula (I), for example, by using HOR.sup.B3 of formula (VIII). In this step, the protected derivative of compounds of formula (VIII), HOR.sup.B3P of formula (IX), may also be used. For example, when R.sup.B3 is CH.sub.2CH.sub.2COOH, the compound of formula (I) wherein R.sup.B is OH may be coupled with an ester of the form HOCH.sub.2CH.sub.2CO.sub.2P, wherein P is a carboxylic acid protecting group as defined herein, followed by removal of the protecting group.

[0113] Compounds of formula (I) wherein

##STR00018##

represents

##STR00019##

may be obtained by isomerisation of compounds of formula (I) wherein

##STR00020##

represents

##STR00021##

under basic conditions, such as those described in the Examples.

##STR00022##

wherein R.sup.A and R.sup.B3 are defined elsewhere herein.

[0114] Step 1: Esterification of commercially available carboxylic acids (VIII) with alcohols of formula (IV) using standard coupling conditions such as DCC and DMAP in a solvent such as DCM, provides compounds of formula (I).

##STR00023##

wherein R.sup.A and R.sup.B3 are defined elsewhere herein, and R.sup.B3P is a protected derivative of R.sup.B3. For example, when R.sup.B3 comprises a carboxylic acid, the acid group may be protected with a carboxylic acid protecting group such as C.sub.1-6 alkyl, e.g., tert-butyl, or para-methoxybenzyl, and when R.sup.B3 comprises a tetrazolyl protecting group, the tetrazolyl group may be protected using a tetrazolyl protecting group such as para-methoxybenzyl or trityl.

[0115] Step 1: Lewis acid catalysed ring opening of itaconic anhydride using alcohols of formula (IX) provides ,-unsaturated carboxylic acids (X).

[0116] Step 2: Esterification of carboxylic acids (X) with alcohols (IV) under standard conditions (such as DCC, DMAP in DCM) provides compounds of formula (I).

[0117] When R.sup.B comprises CH.sub.2COOH or CH.sub.2CH.sub.2COOH, suitably the carboxylic acid protecting group replaces the hydrogen atom attached to an oxygen atom, i.e., R.sup.B3P is CH.sub.2COOP or CH.sub.2CH.sub.2COOP, wherein P is the protecting group

[0118] When R.sup.B comprises CH.sub.2tetrazolyl or CH.sub.2CH.sub.2tetrazolyl, suitably the tetrazolyl protecting group replaces the hydrogen atom attached to a nitrogen atom:

##STR00024##

i.e., R.SUP.B3P. is

[0119] ##STR00025##

wherein the dashed line indicates attachment to the remainder of the compound of formula (I), and P is the protecting group.

[0120] If no R.sup.B3 protecting group is required, for example, when R.sup.B3 is C.sub.1-4 alkyl or C.sub.1-4 haloalkyl, the synthetic route in Scheme 3 may be followed, using a compound of formula HOR.sup.B3 (VIII) in step 1, instead of HO-R.sup.B3P

[0121] Thus, in one embodiment, there is provided a compound of formula (II):

##STR00026## [0122] wherein R.sup.A is defined elsewhere herein and P is a carboxylic acid protecting group such as CH.sub.2CCl.sub.3; [0123] or salts thereof.

[0124] In one embodiment, there is provided a compound of formula (III):

##STR00027## [0125] wherein R.sup.A is defined elsewhere herein and P is a carboxylic acid protecting group such as CH.sub.2CCl.sub.3; [0126] or salts thereof.

[0127] In one embodiment, there is provided a compound of formula (X):

##STR00028## [0128] wherein R.sup.B3P is defined elsewhere herein; [0129] or salts thereof.

[0130] In one embodiment, there is provided a process for the preparation of compounds of formula (I), or salts, such as pharmaceutically acceptable salts, thereof, which comprises the step of deprotecting compounds of formula (III):

##STR00029## [0131] or salts thereof; [0132] wherein R.sup.A is defined elsewhere herein and P is a carboxylic acid protecting group such as CH.sub.2CCl.sub.3.

[0133] It will be appreciated that for use in therapy the salts of the compounds of formula (I) should be pharmaceutically acceptable. Suitable pharmaceutically acceptable salts will be apparent to those skilled in the art. Pharmaceutically acceptable salts include acid addition salts, suitably salts of compounds of the invention comprising a basic group such as an amino group, formed with inorganic acids, e.g., hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid or phosphoric acid. Also included are salts formed with organic acids e.g. succinic acid, maleic acid, acetic acid, fumaric acid, citric acid, tartaric acid, benzoic acid, p-toluenesulfonic acid, methanesulfonic acid, naphthalenesulfonic acid and 1,5-naphthalenedisulfonic acid. Other salts, e.g., oxalates or formates, may be used, for example in the isolation of compounds of formula (I) and are included within the scope of this invention, as are basic addition salts such as sodium, potassium, calcium, aluminium, zinc, magnesium and other metal salts.

[0134] Pharmaceutically acceptable salts may also be formed with organic bases such as basic amines e.g. with ammonia, meglumine, tromethamine, piperazine, arginine, choline, diethylamine, benzathine or lysine. Thus, in one embodiment there is provided a compound of formula (I) in the form of a pharmaceutically acceptable salt. Alternatively, there is provided a compound of formula (I) in the form of a free acid. When the compound contains a basic group as well as the free acid it may be Zwitterionic.

[0135] Suitably, the compound of formula (I) is not a salt e.g. is not a pharmaceutically acceptable salt.

[0136] Suitably, where the compound of formula (I) is in the form of a salt, the pharmaceutically acceptable salt is a basic addition salt such as a carboxylate salt formed with a group 1 metal (e.g. a sodium or potassium salt), a group 2 metal (e.g. a magnesium or calcium salt) or an ammonium salt of a basic amine (e.g. an NH.sub.4 salt), such as a sodium salt.

[0137] The compounds of formula (I) may be prepared in crystalline or non-crystalline form and, if crystalline, may optionally be solvated, e.g. as the hydrate. This invention includes within its scope stoichiometric solvates (e.g. hydrates) as well as compounds containing variable amounts of solvent (e.g. water). Suitably, the compound of formula (I) is not a solvate.

[0138] The invention extends to a pharmaceutically acceptable derivative thereof, such as a pharmaceutically acceptable prodrug of compounds of formula (I). Typical prodrugs of compounds of formula (I) which comprise a carboxylic acid include ester (e.g. C.sub.1-6 alkyl e.g. C.sub.1-4 alkyl ester) derivatives thereof. Thus, in one embodiment, the compound of formula (I) is provided as a pharmaceutically acceptable prodrug. In another embodiment, the compound of formula (I) is not provided as a pharmaceutically acceptable prodrug.

[0139] It is to be understood that the present invention encompasses all isomers of compounds of formula (I) including all geometric, tautomeric and optical forms, and mixtures thereof (e.g. racemic mixtures). Where additional chiral centres are present in compounds of formula (I), the present invention includes within its scope all possible diastereoisomers, including mixtures thereof. The different isomeric forms may be separated or resolved one from the other by conventional methods, or any given isomer may be obtained by conventional synthetic methods or by stereospecific or asymmetric syntheses.

[0140] The present invention also includes all isotopic forms of the compounds provided herein, whether in a form (i) wherein all atoms of a given atomic number have a mass number (or mixture of mass numbers) which predominates in nature (referred to herein as the natural isotopic form) or (ii) wherein one or more atoms are replaced by atoms having the same atomic number, but a mass number different from the mass number of atoms which predominates in nature (referred to herein as an unnatural variant isotopic form). It is understood that an atom may naturally exist as a mixture of mass numbers. The term unnatural variant isotopic form also includes embodiments in which the proportion of an atom of given atomic number having a mass number found less commonly in nature (referred to herein as an uncommon isotope) has been increased relative to that which is naturally occurring e.g. to the level of >20%, >50%, >75%, >90%, >95% or >99% by number of the atoms of that atomic number (the latter embodiment referred to as an isotopically enriched variant form). The term unnatural variant isotopic form also includes embodiments in which the proportion of an uncommon isotope has been reduced relative to that which is naturally occurring. Isotopic forms may include radioactive forms (i.e. they incorporate radioisotopes) and non-radioactive forms. Radioactive forms will typically be isotopically enriched variant forms.

[0141] An unnatural variant isotopic form of a compound may thus contain one or more artificial or uncommon isotopes such as deuterium (.sup.2H or D), carbon-11 (.sup.11C), carbon-13 (.sup.13C), carbon-14 (.sup.14C), nitrogen-13 (.sup.13N), nitrogen-15 (.sup.15N), oxygen-15 (.sup.15O), oxygen-17 (.sup.17O), oxygen-18 (.sup.18O), phosphorus-32 (.sup.32P), sulphur-35 (.sup.35S), chlorine-36 (.sup.36Cl), chlorine-37 (.sup.37Cl), fluorine-18 (.sup.18F) iodine-123 (.sup.123I), iodine-125 (.sup.125I) in one or more atoms or may contain an increased proportion of said isotopes as compared with the proportion that predominates in nature in one or more atoms.

[0142] Unnatural variant isotopic forms comprising radioisotopes may, for example, be used for drug and/or substrate tissue distribution studies. The radioactive isotopes tritium, i.e. .sup.3H, and carbon-14, i.e. .sup.14C, are particularly useful for this purpose in view of their ease of incorporation and ready means of detection. Unnatural variant isotopic forms which incorporate deuterium i.e. .sup.2H or D may afford certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and hence may be preferred in some circumstances. Further, unnatural variant isotopic forms may be prepared which incorporate positron emitting isotopes, such as .sup.11C, .sup.18F, .sup.15O and .sup.13N, and would be useful in positron emission topography (PET) studies for examining substrate receptor occupancy.

[0143] In one embodiment, the compounds of formula (I) are provided in a natural isotopic form. In one embodiment, the compounds of formula (I) are provided in an unnatural variant isotopic form. In a specific embodiment, the unnatural variant isotopic form is a form in which deuterium (i.e. .sup.2H or D) is incorporated where hydrogen is specified in the chemical structure in one or more atoms of a compound of formula (I). In one embodiment, the atoms of the compounds of formula (I) are in an isotopic form which is not radioactive. In one embodiment, one or more atoms of the compounds of formula (I) are in an isotopic form which is radioactive. Suitably radioactive isotopes are stable isotopes. Suitably the unnatural variant isotopic form is a pharmaceutically acceptable form.

[0144] In one embodiment, a compound of formula (I) is provided whereby a single atom of the compound exists in an unnatural variant isotopic form. In another embodiment, a compound of formula (I) is provided whereby two or more atoms exist in an unnatural variant isotopic form.

[0145] Unnatural isotopic variant forms can generally be prepared by conventional techniques known to those skilled in the art or by processes described herein, e.g., processes analogous to those described in the accompanying Examples for preparing natural isotopic forms. Thus, unnatural isotopic variant forms could be prepared by using appropriate isotopically variant (or labelled) reagents in place of the normal reagents employed in the Examples. Since the compounds of formula (I) are intended for use in pharmaceutical compositions it will readily be understood that they are each preferably provided in substantially pure form, for example at least 60% pure, more suitably at least 75% pure and preferably at least 85%, especially at least 98% pure (% are on a weight for weight basis). Impure preparations of the compounds may be used for preparing the more pure forms used in the pharmaceutical compositions.

Therapeutic Indications

[0146] Compounds of formula (I) are of use in therapy, particularly for treating or preventing an inflammatory disease or a disease associated with an undesirable immune response. As shown in Biological Example 1 below, example compounds of formula (I) reduced cytokine release more effectively than dimethyl itaconate and 1-monomethyl itaconate as demonstrated by lower IC.sub.50 values. Cytokines are important mediators of inflammation and immune-mediated disease as evidenced by the therapeutic benefit delivered by antibodies targeting them.

[0147] Thus, in a first aspect, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein, for use as a medicament. Also provided is a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein. Such a pharmaceutical composition contains the compound of formula (I) and a pharmaceutically acceptable carrier or excipient.

[0148] In a further aspect, the present invention provides a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein, for use in treating or preventing an inflammatory disease or a disease associated with an undesirable immune response. In a further aspect, the present invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein, in the manufacture of a medicament for treating or preventing an inflammatory disease or a disease associated with an undesirable immune response. In a further aspect, the present invention provides a method of treating or preventing an inflammatory disease or a disease associated with an undesirable immune response, which comprises administering a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein.

[0149] For all aspects of the invention, suitably the compound is administered to a subject in need thereof, wherein the subject is suitably a human subject.

[0150] In one embodiment is provided a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein, for use in treating an inflammatory disease or disease associated with an undesirable immune response. In one embodiment of the invention is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein, in the manufacture of a medicament for treating an inflammatory disease or a disease associated with an undesirable immune response. In one embodiment of the invention is provided a method of treating an inflammatory disease or a disease associated with an undesirable immune response, which comprises administering a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein.

[0151] In one embodiment is provided a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein, for use in preventing an inflammatory disease or a disease associated with an undesirable immune response. In one embodiment of the invention is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein, in the manufacture of a medicament for preventing an inflammatory disease or a disease associated with an undesirable immune response. In one embodiment of the invention is provided a method of preventing an inflammatory disease or a disease associated with an undesirable immune response, which comprises administering a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein.

[0152] In one embodiment is provided a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein, for use in treating or preventing an inflammatory disease. In one embodiment of the invention is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein, in the manufacture of a medicament for treating or preventing an inflammatory disease. In one embodiment of the invention is provided a method of treating or preventing an inflammatory disease, which comprises administering a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein.

[0153] In one embodiment is provided a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein, for use in treating or preventing a disease associated with an undesirable immune response. In one embodiment of the invention is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein, in the manufacture of a medicament for treating or preventing a disease associated with an undesirable immune response. In one embodiment of the invention is provided a method of treating or preventing a disease associated with an undesirable immune response, which comprises administering a compound of formula (I) or a pharmaceutically acceptable salt and/or solvate thereof as defined herein.

[0154] An undesirable immune response will typically be an immune response which gives rise to a pathology i.e. is a pathological immune response or reaction.

[0155] In one embodiment, the inflammatory disease or disease associated with an undesirable immune response is an auto-immune disease.

[0156] In one embodiment, the inflammatory disease or disease associated with an undesirable immune response is, or is associated with, a disease selected from the group consisting of: psoriasis (including chronic plaque, erythrodermic, pustular, guttate, inverse and nail variants), asthma, chronic obstructive pulmonary disease (COPD, including chronic bronchitis and emphysema), heart failure (including left ventricular failure), myocardial infarction, angina pectoris, other atherosclerosis and/or atherothrombosis-related disorders (including peripheral vascular disease and ischaemic stroke), a mitochondrial and neurodegenerative disease (such as Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, retinitis pigmentosa or mitochondrial encephalomyopathy), autoimmune paraneoplastic retinopathy, transplantation rejection (including antibody-mediated and T cell-mediated forms), multiple sclerosis, transverse myelitis, ischaemia-reperfusion injury (e.g. during elective surgery such as cardiopulmonary bypass for coronary artery bypass grafting or other cardiac surgery, following percutaneous coronary intervention, following treatment of acute ST-elevation myocardial infarction or ischaemic stroke, organ transplantation, or acute compartment syndrome), AGE-induced genome damage, an inflammatory bowel disease (e.g. Crohn's disease or ulcerative colitis), primary sclerosing cholangitis (PSC), PSC-autoimmune hepatitis overlap syndrome, non-alcoholic fatty liver disease (non-alcoholic steatohepatitis), rheumatica, granuloma annulare, cutaneous lupus erythematosus (CLE), systemic lupus erythematosus (SLE), lupus nephritis, drug-induced lupus, autoimmune myocarditis or myopericarditis, Dressler's syndrome, giant cell myocarditis, post-pericardiotomy syndrome, drug-induced hypersensitivity syndromes (including hypersensitivity myocarditis), eczema, sarcoidosis, erythema nodosum, acute disseminated encephalomyelitis (ADEM), neuromyelitis optica spectrum disorders, MOG (myelin oligodendrocyte glycoprotein) antibody-associated disorders (including MOG-EM), optic neuritis, CLIPPERS (chronic lymphocytic inflammation with pontine perivascular enhancement responsive to steroids), diffuse myelinoclastic sclerosis, Addison's disease, alopecia areata, ankylosing spondylitis, other spondyloarthritides (including peripheral spondyloarthritis, that is associated with psoriasis, inflammatory bowel disease, reactive arthritis or juvenile onset forms), antiphospholipid antibody syndrome, autoimmune hemolytic anaemia, autoimmune hepatitis, autoimmune inner ear disease, pemphigoid (including bullous pemphigoid, mucous membrane pemphigoid, cicatricial pemphigoid, herpes gestationis or pemphigoid gestationis, ocular cicatricial pemphigoid), linear IgA disease, Behget's disease, celiac disease, Chagas disease, dermatomyositis, diabetes mellitus type I, endometriosis, Goodpasture's syndrome, Graves' disease, Guillain-Barre syndrome and its subtypes (including acute inflammatory demyelinating polyneuropathy, AIDP, acute motor axonal neuropathy (AMAN), acute motor and sensory axonal neuropathy (AMSAN), pharyngeal-cervical-brachial variant, Miller-Fisher variant and Bickerstaff's brainstem encephalitis), progressive inflammatory neuropathy, Hashimoto's disease, hidradenitis suppurativa, inclusion body myositis, necrotising myopathy, Kawasaki disease, IgA nephropathy, Henoch-Schonlein purpura, idiopathic thrombocytopenic purpura, thrombotic thrombocytopenic purpura (TTP), Evans' syndrome, interstitial cystitis, mixed connective tissue disease, undifferentiated connective tissue disease, morphea, myasthenia gravis (including MuSK antibody positive and seronegative variants), narcolepsy, neuromyotonia, pemphigus vulgaris, pernicious anaemia, psoriatic arthritis, polymyositis, primary biliary cholangitis (also known as primary biliary cirrhosis), rheumatoid arthritis, palindromic rheumatism, schizophrenia, autoimmune (meningo-)encephalitis syndromes, scleroderma, Sjogren's syndrome, stiff person syndrome, polymylagia rheumatica, giant cell arteritis (temporal arteritis), Takayasu arteritis, polyarteritis nodosa, Kawasaki disease, granulomatosis with polyangitis (GPA; formerly known as Wegener's granulomatosis), eosinophilic granulomatosis with polyangiitis (EGPA; formerly known as Churg-Strauss syndrome), microscopic polyarteritis/polyangiitis, hypocomplementaemic urticarial vasculitis, hypersensitivity vasculitis, cryoglobulinemia, thromboangiitis obliterans (Buerger's disease), vasculitis, leukocytoclastic vasculitis, vitiligo, acute disseminated encephalomyelitis, adrenoleukodystrophy, Alexander's disease, Alper's disease, balo concentric sclerosis or Marburg disease, cryptogenic organising pneumonia (formerly known as bronchiolitis obliterans organizing pneumonia), Canavan disease, central nervous system vasculitic syndrome, Charcot-Marie-Tooth disease, childhood ataxia with central nervous system hypomyelination, chronic inflammatory demyelinating polyneuropathy (CIDP), diabetic retinopathy, globoid cell leukodystrophy (Krabbe disease), graft-versus-host disease (GVHD) (including acute and chronic forms, as well as intestinal GVHD), hepatitis C (HCV) infection or complication, herpes simplex viral infection or complication, human immunodeficiency virus (HIV) infection or complication, lichen planus, monomelic amyotrophy, cystic fibrosis, pulmonary arterial hypertension (PAH, including idiopathic PAH), lung sarcoidosis, idiopathic pulmonary fibrosis, paediatric asthma, atopic dermatitis, allergic dermatitis, contact dermatitis, allergic rhinitis, rhinitis, sinusitis, conjunctivitis, allergic conjunctivitis, keratoconjunctivitis sicca, dry eye, xerophthalmia, glaucoma, macular oedema, diabetic macular oedema, central retinal vein occlusion (CRVO), macular degeneration (including dry and/or wet age related macular degeneration, AMD), post-operative cataract inflammation, uveitis (including posterior, anterior, intermediate and pan uveitis), iridocyclitis, scleritis, corneal graft and limbal cell transplant rejection, gluten sensitive enteropathy (coeliac disease), dermatitis herpetiformis, eosinophilic esophagitis, achalasia, autoimmune dysautonomia, autoimmune encephalomyelitis, autoimmune oophoritis, autoimmune orchitis, autoimmune pancreatitis, aortitis and periaortitis, autoimmune retinopathy, autoimmune urticaria, (idiopathic) Castleman's disease, Cogan's syndrome, IgG4-related disease, retroperitoneal fibrosis, juvenile idiopathic arthritis including systemic juvenile idiopathic arthritis (Still's disease), adult-onset Still's disease, ligneous conjunctivitis, Mooren's ulcer, pityriasis lichenoides et varioliformis acuta (PLEVA, also known as Mucha-Habermann disease), multifocal motor neuropathy (MMN), paediatric acute-onset neuropsychiatric syndrome (PANS) (including paediatric autoimmune neuropsychiatric disorders associated with streptococcal infections (PANDAS)), paraneoplastic syndromes (including paraneoplastic cerebellar degeneration, Lambert-Eaton myaesthenic syndrome, limbic encephalitis, brainstem encephalitis, opsoclonus myoclonus ataxia syndrome, anti-NMDA receptor encephalitis, thymoma-associated multiorgan autoimmunity), perivenous encephalomyelitis, reflex sympathetic dystrophy, relapsing polychondritis, sperm & testicular autoimmunity, Susac's syndrome, Tolosa-Hunt syndrome, Vogt-Koyanagi-Harada Disease, anti-synthetase syndrome, autoimmune enteropathy, immune dysregulation polyendocrinopathy enteropathy X-linked (IPEX), microscopic colitis, autoimmune lymphoproliferative syndrome (ALPS), autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy syndrome (APEX), gout, pseudogout, amyloid (including AA or secondary amyloidosis), eosinophilic fasciitis (Shulman syndrome) progesterone hypersensitivity (including progesterone dermatitis), familial Mediterranean fever (FMF), tumour necrosis factor (TNF) receptor-associated periodic fever syndrome (TRAPS), hyperimmunoglobulinaemia D with periodic fever syndrome (HIDS), PAPA (pyogenic arthritis, pyoderma gangrenosum, severe cystic acne) syndrome, deficiency of interleukin-1 receptor antagonist (DIRA), deficiency of the interleukin-36-receptor antagonist (DITRA), cryopyrin-associated periodic syndromes (CAPS) (including familial cold autoinflammatory syndrome [FCAS], Muckle-Wells syndrome, neonatal onset multisystem inflammatory disease [NOMID]), NLRP12-associated autoinflammatory disorders (NLRP12AD), periodic fever aphthous stomatitis (PFAPA), chronic atypical neutrophilic dermatosis with lipodystrophy and elevated temperature (CANDLE), Majeed syndrome, Blau syndrome (also known as juvenile systemic granulomatosis), macrophage activation syndrome, chronic recurrent multifocal osteomyelitis (CRMO), familial cold autoinflammatory syndrome, mutant adenosine deaminase 2 and monogenic interferonopathies (including Aicardi-Goutieres syndrome, retinal vasculopathy with cerebral leukodystrophy, spondyloenchondrodysplasia, STING [stimulator of interferon genes]-associated vasculopathy with onset in infancy, proteasome associated autoinflammatory syndromes, familial chilblain lupus, dyschromatosis symmetrica hereditaria), Schnitzler syndrome; familial cylindromatosis, congenital B cell lymphocytosis, OTULIN-related autoinflammatory syndrome, type 2 diabetes mellitus, insulin resistance and the metabolic syndrome (including obesity-associated inflammation), atherosclerotic disorders (e.g. myocardial infarction, angina, ischaemic heart failure, ischaemic nephropathy, ischaemic stroke, peripheral vascular disease, aortic aneurysm), and renal inflammatory disorders (e.g. diabetic nephropathy, membranous nephropathy, minimal change disease, crescentic glomerulonephritis, acute kidney injury, renal transplantation).

[0157] In one embodiment, the inflammatory disease or disease associated with an undesirable immune response is, or is associated with, a disease selected from the following autoinflammatory diseases: familial Mediterranean fever (FMF), tumour necrosis factor (TNF) receptor-associated periodic fever syndrome (TRAPS), hyperimmunoglobulinaemia D with periodic fever syndrome (HIDS), PAPA (pyogenic arthritis, pyoderma gangrenosum, and severe cystic acne) syndrome, deficiency of interleukin-1 receptor antagonist (DIRA), deficiency of the interleukin-36-receptor antagonist (DITRA), cryopyrin-associated periodic syndromes (CAPS) (including familial cold autoinflammatory syndrome [FCAS], Muckle-Wells syndrome, and neonatal onset multisystem inflammatory disease [NOMID]), NLRP12-associated autoinflammatory disorders (NLRP12AD), periodic fever aphthous stomatitis (PFAPA), chronic atypical neutrophilic dermatosis with lipodystrophy and elevated temperature (CANDLE), Majeed syndrome, Blau syndrome (also known as juvenile systemic granulomatosis), macrophage activation syndrome, chronic recurrent multifocal osteomyelitis (CRMO), familial cold autoinflammatory syndrome, mutant adenosine deaminase 2 and monogenic interferonopathies (including Aicardi-Goutieres syndrome, retinal vasculopathy with cerebral leukodystrophy, spondyloenchondrodysplasia, STING [stimulator of interferon genes]-associated vasculopathy with onset in infancy, proteasome associated autoinflammatory syndromes, familial chilblain lupus, dyschromatosis symmetrica hereditaria) and Schnitzler syndrome.

[0158] In one embodiment, the inflammatory disease or disease associated with an undesirable immune response is, or is associated with, a disease selected from the following diseases mediated by excess NF-B or gain of function in the NF-B signalling pathway or in which there is a major contribution to the abnormal pathogenesis therefrom (including non-canonical NF-B signalling): familial cylindromatosis, congenital B cell lymphocytosis, OTULIN-related autoinflammatory syndrome, type 2 diabetes mellitus, insulin resistance and the metabolic syndrome (including obesity-associated inflammation), atherosclerotic disorders (e.g. myocardial infarction, angina, ischaemic heart failure, ischaemic nephropathy, ischaemic stroke, peripheral vascular disease, aortic aneurysm), renal inflammatory disorders (e.g. diabetic nephropathy, membranous nephropathy, minimal change disease, crescentic glomerulonephritis, acute kidney injury, renal transplantation), asthma, COPD, type 1 diabetes mellitus, rheumatoid arthritis, multiple sclerosis, inflammatory bowel disease (including ulcerative colitis and Crohn's disease), and SLE.

[0159] In one embodiment, the disease is selected from the group consisting of rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, systemic lupus erythematosus, multiple sclerosis, psoriasis, Crohn's disease, ulcerative colitis, uveitis, cryopyrin-associated periodic syndromes, Muckle-Wells syndrome, juvenile idiopathic arthritis, chronic obstructive pulmonary disease and asthma.

[0160] In one embodiment, the disease is multiple sclerosis.

[0161] In one embodiment, the disease is psoriasis.

[0162] In one embodiment, the disease is asthma.

[0163] In one embodiment, the disease is chronic obstructive pulmonary disease.

[0164] In one embodiment, the disease is systemic lupus erythematosus.

Administration

[0165] The compound of formula (I) is usually administered as a pharmaceutical composition. Thus, in one embodiment, is provided a pharmaceutical composition comprising a compound of formula (I) and one or more pharmaceutically acceptable diluents or carriers.

[0166] The compound of formula (I) may be administered by any convenient method, e.g. by oral, parenteral, buccal, sublingual, nasal, rectal, intrathecal or transdermal administration, and the pharmaceutical compositions adapted accordingly.

[0167] The compound of formula (I) may be administered topically to the target organ e.g. topically to the eye, lung, nose or skin. Hence the invention provides a pharmaceutical composition comprising a compound of formula (I) optionally in combination with one or more topically acceptable diluents or carriers.

[0168] A compound of formula (I) which is active when given orally can be formulated as a liquid or solid, e.g. as a syrup, suspension, emulsion, tablet, capsule or lozenge.

[0169] A liquid formulation will generally consist of a suspension or solution of the compound of formula (I) in a suitable liquid carrier(s). Suitably the carrier is non-aqueous e.g. polyethylene glycol or an oil. The formulation may also contain a suspending agent, preservative, flavouring and/or colouring agent.

[0170] A composition in the form of a tablet can be prepared using any suitable pharmaceutical carrier(s) routinely used for preparing solid formulations, such as magnesium stearate, starch, lactose, sucrose and cellulose.

[0171] A composition in the form of a capsule can be prepared using routine encapsulation procedures, e.g. pellets containing the active ingredient can be prepared using standard carriers and then filled into a hard gelatine capsule; alternatively, a dispersion or suspension can be prepared using any suitable pharmaceutical carrier(s), e.g. aqueous gums, celluloses, silicates or oils and the dispersion or suspension then filled into a soft gelatine capsule.

[0172] Typical parenteral compositions consist of a solution or suspension of the compound of formula (I) in a sterile aqueous carrier or parenterally acceptable oil, e.g. polyethylene glycol, polyvinyl pyrrolidone, lecithin, arachis oil or sesame oil. Alternatively, the solution can be lyophilised and then reconstituted with a suitable solvent just prior to administration.

[0173] Compositions for nasal administration may conveniently be formulated as aerosols, drops, gels and powders. Aerosol formulations typically comprise a solution or fine suspension of the compound of formula (I) in a pharmaceutically acceptable aqueous or non-aqueous solvent and are usually presented in single or multidose quantities in sterile form in a sealed container which can take the form of a cartridge or refill for use with an atomising device. Alternatively, the sealed container may be a disposable dispensing device such as a single dose nasal inhaler or an aerosol dispenser fitted with a metering valve. Where the dosage form comprises an aerosol dispenser, it will contain a propellant which can be a compressed gas e.g. air, or an organic propellant such as a chlorofluorocarbon (CFC) or a hydrofluorocarbon (HFC). Aerosol dosage forms can also take the form of pump-atomisers.

[0174] Topical administration to the lung may be achieved by use of an aerosol formulation. Aerosol formulations typically comprise the active ingredient suspended or dissolved in a suitable aerosol propellant, such as a chlorofluorocarbon (CFC) or a hydrofluorocarbon (HFC).

[0175] Topical administration to the lung may also be achieved by use of a non-pressurised formulation such as an aqueous solution or suspension. These may be administered by means of a nebuliser e.g. one that can be hand-held and portable or for home or hospital use (i.e. non-portable). The formulation may comprise excipients such as water, buffers, tonicity adjusting agents, pH adjusting agents, surfactants and co-solvents.

[0176] Topical administration to the lung may also be achieved by use of a dry-powder formulation. The formulation will typically contain a topically acceptable diluent such as lactose, glucose or mannitol (preferably lactose).

[0177] The compound of the invention may also be administered rectally, for example in the form of suppositories or enemas, which include aqueous or oily solutions as well as suspensions and emulsions and foams. Such compositions are prepared following standard procedures, well known by those skilled in the art. For example, suppositories can be prepared by mixing the active ingredient with a conventional suppository base such as cocoa butter or other glycerides.

[0178] In this case, the drug is mixed with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols.

[0179] Generally, for compositions intended to be administered topically to the eye in the form of eye drops or eye ointments, the total amount of the compound of the present invention will be about 0.0001 to less than 4.0% (w/w).

[0180] Preferably, for topical ocular administration, the compositions administered according to the present invention will be formulated as solutions, suspensions, emulsions and other dosage forms.

[0181] The compositions administered according to the present invention may also include various other ingredients, including, but not limited to, tonicity agents, buffers, surfactants, stabilizing polymer, preservatives, co-solvents and viscosity building agents. Suitable pharmaceutical compositions of the present invention include a compound of the invention formulated with a tonicity agent and a buffer. The pharmaceutical compositions of the present invention may further optionally include a surfactant and/or a palliative agent and/or a stabilizing polymer.

[0182] Various tonicity agents may be employed to adjust the tonicity of the composition, preferably to that of natural tears for ophthalmic compositions. For example, sodium chloride, potassium chloride, magnesium chloride, calcium chloride, simple sugars such as dextrose, fructose, galactose, and/or simply polyols such as the sugar alcohols mannitol, sorbitol, xylitol, lactitol, isomaltitol, maltitol, and hydrogenated starch hydrolysates may be added to the composition to approximate physiological tonicity. Such an amount of tonicity agent will vary, depending on the particular agent to be added. In general, however, the compositions will have a tonicity agent in an amount sufficient to cause the final composition to have an ophthalmically acceptable osmolality (generally about 150-450 mOsm, preferably 250-350 mOsm and most preferably at approximately 290 mOsm). In general, the tonicity agents of the invention will be present in the range of 2 to 4% w/w. Preferred tonicity agents of the invention include the simple sugars or the sugar alcohols, such as D-mannitol.

[0183] An appropriate buffer system (e.g. sodium phosphate, sodium acetate, sodium citrate, sodium borate or boric acid) may be added to the compositions to prevent pH drift under storage conditions. The particular concentration will vary, depending on the agent employed. Preferably however, the buffer will be chosen to maintain a target pH within the range of pH 5 to 8, and more preferably to a target pH of pH 5 to 7.

[0184] Surfactants may optionally be employed to deliver higher concentrations of compound of the present invention. The surfactants function to solubilise the compound and stabilise colloid dispersion, such as micellar solution, microemulsion, emulsion and suspension. Examples of surfactants which may optionally be used include polysorbate, poloxamer, polyosyl 40 stearate, polyoxyl castor oil, tyloxapol, Triton, and sorbitan monolaurate. Preferred surfactants to be employed in the invention have a hydrophile/lipophile/balance HLB in the range of 12.4 to 13.2 and are acceptable for ophthalmic use, such as TritonX114 and tyloxapol.

[0185] Additional agents that may be added to the ophthalmic compositions of compounds of the present invention are demulcents which function as a stabilising polymer. The stabilizing polymer should be an ionic/charged example with precedence for topical ocular use, more specifically, a polymer that carries negative charge on its surface that can exhibit a zeta-potential of ()10-50 mV for physical stability and capable of making a dispersion in water (i.e. water soluble). A preferred stabilising polymer of the invention would be polyelectrolyte, or polyelectrolytes if more than one, from the family of cross-linked polyacrylates, such as carbomers and Pemulen, specifically Carbomer 974p (polyacrylic acid), at 0.1-0.5% w/w.

[0186] Other compounds may also be added to the ophthalmic compositions of the compound of the present invention to increase the viscosity of the carrier. Examples of viscosity enhancing agents include, but are not limited to: polysaccharides, such as hyaluronic acid and its salts, chondroitin sulfate and its salts, dextrans, various polymers of the cellulose family; vinyl polymers; and acrylic acid polymers.

[0187] Topical ophthalmic products are typically packaged in multidose form. Preservatives are thus required to prevent microbial contamination during use. Suitable preservatives include: benzalkonium chloride, chlorobutanol, benzododecinium bromide, methyl paraben, propyl paraben, phenylethyl alcohol, edentate disodium, sorbic acid, polyquaternium-1, or other agents known to those skilled in the art. Such preservatives are typically employed at a level of from 0.001 to 1.0% w/v. Unit dose compositions of the present invention will be sterile, but typically unpreserved. Such compositions, therefore, generally will not contain preservatives.

[0188] Compositions suitable for buccal or sublingual administration include tablets, lozenges and pastilles where the compound of formula (I) is formulated with a carrier such as sugar and acacia, tragacanth, or gelatine and glycerine.

[0189] Compositions suitable for transdermal administration include ointments, gels and patches.

[0190] The composition may contain from 0.1% to 100% by weight, for example from 10% to 60% by weight, of the compound of formula (I), depending on the method of administration. The composition may contain from 0% to 99% by weight, for example 40% to 90% by weight, of the carrier, depending on the method of administration. The composition may contain from 0.05 mg to 1000 mg, for example from 1.0 mg to 500 mg, such as from 1.0 mg to 50 mg, e.g. about 10 mg of the compound of formula (I), depending on the method of administration. The composition may contain from 50 mg to 1000 mg, for example from 100 mg to 400 mg of the carrier, depending on the method of administration. The dose of the compound used in the treatment of the aforementioned disorders will vary in the usual way with the seriousness of the disorders, the weight of the sufferer, and other similar factors. However, as a general guide suitable unit doses may be 0.05 mg to 1000 mg, more suitably 1.0 mg to 500 mg, such as from 1.0 mg to 50 mg, e.g. about 10 mg and such unit doses may be administered more than once a day, for example two or three times a day. Such therapy may extend for a number of weeks or months.

[0191] In one embodiment of the invention, the compound of formula (I) is used in combination with a further therapeutic agent or agents. When the compound of formula (I) is used in combination with other therapeutic agents, the compounds may be administered either sequentially or simultaneously by any convenient route. Alternatively, the compounds may be administered separately.

[0192] Therapeutic agents which may be used in combination with the present invention include: corticosteroids (glucocorticoids), retinoids (e.g. acitretin, isotretinoin, tazarotene), anthralin, vitamin D analogues (e.g. cacitriol, calcipotriol), calcineurin inhibitors (e.g. tacrolimus, pimecrolimus), phototherapy or photochemotherapy (e.g. psoralen ultraviolet irradiation, PUVA) or other form of ultraviolet light irradiation therapy, ciclosporine, thiopurines (e.g. azathioprine, 6-mercaptopurine), methotrexate, anti-TNF agents (e.g. infliximab, etanercept, adalimumab, certolizumab, golimumab and biosimilars), phosphodiesterase-4 (PDE4) inhibition (e.g. apremilast, crisaborole), anti-IL-17 agents (e.g. brodalumab, ixekizumab, secukinumab), anti-IL12/IL-23 agents (e.g. ustekinumab, briakinumab), anti-IL-23 agents (e.g. guselkumab, tildrakizumab), JAK (Janus Kinase) inhibitors (e.g. tofacitinib, ruxolitinib, baricitinib, filgotinib, upadacitinib), plasma exchange, intravenous immune globulin (IVIG), cyclophosphamide, anti-CD20 B cell depleting agents (e.g. rituximab, ocrelizumab, ofatumumab, obinutuzumab), anthracycline analogues (e.g. mitoxantrone), cladribine, sphingosine 1-phosphate receptor modulators or sphingosine analogues (e.g. fingolimod, siponimod, ozanimod, etrasimod), interferon beta preparations (including interferon beta 1b/1a), glatiramer, anti-CD3 therapy (e.g. OKT3), anti-CD52 targeting agents (e.g. alemtuzumab), leflunomide, teriflunomide, gold compounds, laquinimod, potassium channel blockers (e.g. dalfampridine/4-aminopyridine), mycophenolic acid, mycophenolate mofetil, purine analogues (e.g. pentostatin), mTOR (mechanistic target of rapamycin) pathway inhibitors (e.g. sirolimus, everolimus), anti-thymocyte globulin (ATG), IL-2 receptor (CD25) inhibitors (e.g. basiliximab, daclizumab), anti-IL-6 receptor or anti-IL-6 agents (e.g. tocilizumab, siltuximab), Bruton's tyrosine kinase (BTK) inhibitors (e.g. ibrutinib), tyrosine kinase inhibitors (e.g. imatinib), ursodeoxycholic acid, hydroxychloroquine, chloroquine, B cell activating factor (BAFF, also known as BLyS, B lymphocyte stimulator) inhibitors (e.g. belimumab, blisibimod), other B cell targeted therapy including fusion proteins targeting both APRIL (A PRoliferation-Inducing Ligand) and BLyS (e.g. atacicept), PI3K inhibitors including pan-inhibitors or those targeting the p110 and/or p110 containing isoforms (e.g. idelalisib, copanlisib, duvelisib), interferon receptor inhibitors (e.g. anifrolumab, sifalimumab), T cell co-stimulation blockers (e.g. abatacept, belatacept), thalidomide and its derivatives (e.g. lenalidomide), dapsone, clofazimine, leukotriene antagonists (e.g. montelukast), theophylline, anti-IgE therapy (e.g. omalizumab), anti-IL-5 agents (e.g. mepolizumab, reslizumab), long-acting muscarinic agents (e.g. tiotropium, aclidinium, umeclidinium), PDE4 inhibitors (e.g. roflumilast), riluzole, free radical scavengers (e.g. edaravone), proteasome inhibitors (e.g. bortezomib), complement cascade inhibitors including those directed against C5 (e.g. eculizumab), immunoadsor, antithymocyte globulin, 5-aminosalicylates and their derivatives (e.g. sulfasalazine, balsalazide, mesalamine), anti-integrin agents including those targeting 41 and/or 47 integrins (e.g. natalizumab, vedolizumab), anti-CD11- agents (e.g. efalizumab), non-steroidal anti-inflammatory drugs (NSAIDs) including the salicylates (e.g. aspirin), propionic acids (e.g. ibuprofen, naproxen), acetic acids (e.g. indomethacin, diclofenac, etodolac), oxicams (e.g. meloxicam) and fenamates (e.g. mefenamic acid), selective or relatively selective COX-2 inhibitors (e.g. celecoxib, etroxicoxib, valdecoxib and etodolac, meloxicam, nabumetone), colchicine, IL-4 receptor inhibitors (e.g. dupilumab), topical/contact immunotherapy (e.g. diphenylcyclopropenone, squaric acid dibutyl ester), anti-IL-1 receptor therapy (e.g. anakinra), IL-1 inhibitor (e.g. canakinumab), IL-1 neutralising therapy (e.g. rilonacept), chlorambucil, specific antibiotics with immunomodulatory properties and/or ability to modulate NRF2 (e.g. tetracyclines including minocycline, clindamycin, macrolide antibiotics), anti-androgenic therapy (e.g. cyproterone, spironolactone, finasteride), pentoxifylline, ursodeoxycholic acid, obeticholic acid, fibrate, cystic fibrosis transmembrane conductance regulator (CFTR) modulators, VEGF (vascular endothelial growth factor) inhibitors (e.g. bevacizumab, ranibizumab, pegaptanib, aflibercept), pirfenidone, and mizoribine.

[0193] Compounds of formula (I) may display one or more of the following desirable properties: [0194] low IC.sub.50 values for inhibiting release of cytokines, e.g., IL-1p and/or IL-6, from cells; [0195] low EC.sub.50 and/or high E.sub.max values for activating the NRF2 pathway; [0196] high EC.sub.50 and/or low E.sub.max values for activating the NRF2 pathway; [0197] enhanced efficacy through improved hydrolytic stability and/or augmented maximum response; [0198] reduced dose and dosing frequency through improved pharmacokinetics; [0199] improved oral systemic bioavailability; [0200] reduced plasma clearance following intravenous dosing; [0201] augmented cell permeability; [0202] enhanced aqueous solubility; [0203] good tolerability, for example, by limiting the flushing and/or gastrointestinal side effects provoked by oral DMF (Hunt T. et al., 2015; WO2014/152494A1, incorporated herein by reference), possibly by reducing or eliminating HCA2 activity; [0204] low toxicity at the relevant therapeutic dose; [0205] distinct anti-inflammatory profiles resulting from varied electrophilicities, leading to differential targeting of the cysteine proteome (van der Reest J. et al., 2018) and, therefore, modified effects on gene activation).

Abbreviations

[0206] 4OI 4-octyl itaconic acid [0207] Ac acetyl [0208] aq. aqueous [0209] BBFO broadband fluorine observe [0210] BEH ethylene bridged hybrid [0211] BTEAC benzyl(triethyl)ammonium chloride [0212] Bu butyl [0213] ca. circa [0214] CSH charged surface hybrid [0215] d doublet [0216] DAD diode array detector [0217] DBU 1,8-diazabicyclo(5.4.0)undec-7-ene [0218] DCC N,N-dicyclohexylcarbodiimide [0219] DCM dichloromethane [0220] DIPEA N,N-diisopropylethylamine [0221] DMAP 4-dimethylaminopyridine [0222] DMF dimethyl fumarate [0223] DMSO dimethyl sulfoxide [0224] EDCI 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide [0225] Et ethyl [0226] ES.sup.+ electrospray [0227] FBS fetal bovine serum [0228] g gram(s) [0229] h hour(s) [0230] HATU 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid [0231] hexafluorophosphate [0232] HPLC high-performance liquid chromatography [0233] LCMS liquid chromatography-mass spectrometry [0234] LPS lipopolysaccharide [0235] m multiplet [0236] M molar concentration/molar mass [0237] Me methyl [0238] m/z mass to charge ratio [0239] (M)Hz (mega)hertz [0240] min(s) minute(s) [0241] mL millilitres [0242] mm millimeter [0243] mmol millimole [0244] MS mass spectrometry [0245] MSD mass selective detector [0246] MTBE methyl tertiary-butyl ether [0247] nm nanometre [0248] NMP N-methyl-2-pyrrolidone [0249] NMR nuclear magnetic resonance [0250] NRF2 nuclear factor (erythroid-derived 2)-like 2 [0251] O/N overnight [0252] PDA photodiode array [0253] ppm parts per million [0254] rpm revolutions per minute [0255] RT room temperature [0256] s singlet [0257] sat. saturated [0258] t triplet [0259] T3P propanephosphonic acid anhydride [0260] TEA triethylamine [0261] TFA trifluoroacetic acid [0262] THF tetrahydrofuran [0263] L microlitre [0264] M micromolar [0265] UPLC ultra performance liquid chromatography [0266] VWD variable wavelength detector [0267] wt. weight [0268] C. degrees centigrade

EXAMPLES

Analytical Equipment

[0269] NMR spectra were recorded using a Bruker 400 MHz Avance Ill spectrometer fitted with a BBFO 5 mm probe, or a Bruker 500 MHz Avance Ill HD spectrometer equipped with a Bruker 5 mm SmartProbe. Spectra were measured at 298 K, unless indicated otherwise, and were referenced relative to the solvent resonance. The chemical shifts are reported in parts per million. Data were acquired using Bruker TopSpin software.

[0270] UPLC/MS analysis was carried out on a Waters Acquity UPLC system using either a Waters Acquity CSH C18 or BEH C18 column (2.130 mm) maintained at a temperature of 40 C. and eluted with a linear acetonitrile gradient appropriate for the lipophilicity of the compound over 3 or 10 minutes at a constant flow rate of 0.77 mL/min. The aqueous portion of the mobile phase was either 0.1% Formic Acid (CSH C18 column) or 10 mM Ammonium Bicarbonate (BEH C18 column). LC-UV chromatograms were recorded using a Waters Acquity PDA detector between 210 and 400 nm. Mass spectra were recorded using a Waters Acquity Qda detector with electrospray ionisation switching between positive and negative ion mode. Sample concentration was adjusted to give adequate UV response.

[0271] LCMS analysis was carried out on an Agilent LCMS system using either a Waters Acquity CSH C18 or BEH C18 column (4.630 mm) maintained at a temperature of 40 C. and eluted with a linear acetonitrile gradient appropriate for the lipophilicity of the compound over 4 or 15 minutes at a constant flow rate of 2.5 mL/min. The aqueous portion of the mobile phase was either 0.1% Formic Acid (CSH C18 column) or 10 mM Ammonium Bicarbonate (BEH C18 column). LC-UV chromatograms were recorded using an Agilent VWD or DAD detector at 254 nm. Mass spectra were recorded using an Agilent MSD detector with electrospray ionisation switching between positive and negative ion mode. Sample concentration was adjusted to give adequate UV response.

Commercial Materials

[0272] All starting materials disclosed herein are commercially available, unless otherwise stated. Dimethyl itaconate was purchased from Sigma-Aldrich (product number: 109533). 1-Monomethyl itaconate was purchased from Enamine. 4-Octyl itaconate was purchased from BOC biosciences (product number: B0001-007866).

[0273] Unless otherwise stated all reactions were stirred. Organic solutions were routinely dried over anhydrous sodium or magnesium sulfate.

Intermediate 12-methylene-4-oxo-4-(2,2,2-trichloroethoxy)butanoic acid

[0274] ##STR00030##

[0275] To a mixture of 3-methylenedihydrofuran-2,5-dione (5.0 g, 44.6 mmol) and 2,2,2-trichloroethanol (10.0 g, 66.1 mmol) was added boron trifluoride diethyl etherate (634 mg, 4.46 mmol), and the mixture was allowed to stir at 75 C. for 40 minutes. The reaction mixture was cooled to room temperature, quenched with methanol (2 mL), diluted with EtOAc (50 mL) and water (20 mL), separated and extracted with EtOAc (220 mL). The organic layers were combined and washed with brine and dried. The filtrate was concentrated under reduced pressure, the residue was purified by reversed column chromatography (Column: Boston ODS 120 g Flash; Flow Rate: 40 mL/min; solvent system: MeCN/(10 mmol/L HCl/water); MeCN gradient: 60-80%; collection wavelength: 214 nm). The fractions were concentrated under reduced pressure to remove MeCN, and the residue was lyophilized to give 2-methylene-4-oxo-4-(2,2,2-trichloroethoxy)butanoic acid (9.0 g, 77% yield, purity 80%) as white solid. The solid was triturated in n-hexane (80 mL)/MTBE (8 mL) overnight, recovered by filtration, and dried at 40 C. under reduced pressure to give 2-methylene-4-oxo-4-(2,2,2-trichloroethoxy)butanoic acid (8.0 g, 68% yield). LCMS m/z 283.2 (M+Na)+ (ES+).

Intermediate 21-(4-(trifluoromethyl)phenyl)cyclobutan-1-ol

[0276] ##STR00031##

[0277] n-BuLi solution in hexane (2.5 M, 43.7 mL, 109.2 mmol) was added to a solution of 1-bromo-4-(trifluoromethyl)benzene (22.3 g, 99.5 mmol) in THF (180 mL) at 78 C. and the mixture was stirred at 78 C. for 1 hr. Cyclobutanone (7.6 g, 109.2 mmol) was added, and the mixture was stirred at 78 C. for 5 hrs, then quenched with saturated aqueous NH.sub.4Cl solution (200 mL). The phases were separated and the aqueous layer was extracted with MTBE (280 mL). The combined organic layers were washed with brine, dried and concentrated under reduced pressure. The residue was purified by flash column chromatography on silica (0-14% MTBE/petroleum ether) to give 1-(4-(trifluoromethyl)phenyl)cyclobutan-1-ol (16.5 g, 76.3 mmol, 77% yield) as a yellow oil. .sup.1H NMR (400 MHz, CDC.sub.3) : 7.61 (s, 4H), 2.59-2.48 (m, 2H), 2.43-2.32 (m, 2H), 2.12-1.98 (m, 1H), 1.81-1.66 (m, 1H). One exchangeable proton not observed.

Intermediate 31-(3-fluoro-4-(trifluoromethyl)phenyl)cyclobutan-1-ol

[0278] ##STR00032##

[0279] Prepared from 4-bromo-2-fluoro-1-(trifluoromethyl)benzene and cyclobutanone using a similar procedure to Intermediate 2. Obtained as yellow oil (3.65 g, 54% yield). .sup.1H NMR (400 MHz, CDCl.sub.3) : 7.60 (t, J=8.0 Hz, 1H), 7.40-7.35 (m, 2H), 2.57-2.50 (m, 2H), 2.44-2.37 (m, 2H), 2.11-2.09 (m, 1H), 1.80-1.77 (m, 1H).

Intermediate 43-(4-(trifluoromethyl)phenyl)oxetan-3-ol

[0280] ##STR00033##

[0281] Prepared from 1-bromo-4-(trifluoromethyl)benzene and oxetan-3-one using a similar procedure to Intermediate 2. Obtained as yellow oil (700 mg, 58% yield). .sup.1H NMR (400 MHz, CDCl.sub.3) : 7.78 (d, J=8.4 Hz, 2H), 7.69 (d, J=8.4 Hz, 2H), 4.95 (d, J=7.6 Hz, 2H), 4.89 (d, J=7.6 Hz, 2H).

Intermediate 51-(5-(trifluoromethyl)pyridin-2-yl)cyclobutan-1-ol

[0282] ##STR00034##

[0283] Prepared from 2-bromo-5-(trifluoromethyl)pyridine and cyclobutanone using a similar procedure to Intermediate 2. Obtained as yellow oil (250 mg, 65% yield). .sup.1H NMR (400 MHz, CDCl.sub.3) : 8.81 (s, 1H), 7.99 (dd, J=8.3, 2.3 Hz, 1H), 7.72 (d, J=8.3 Hz, 1H), 4.73 (s, 1H), 2.61-2.49 (m, 4H), 2.17-2.07 (m, 1H), 1.98-1.86 (m, 1H).

Intermediate 61-(5-(trifluoromethyl)thiophen-2-yl)cyclobutan-1-ol

[0284] ##STR00035##

[0285] Prepared from 2-bromo-5-(trifluoromethyl)thiophene and cyclobutanone using a similar procedure to Intermediate 2. Obtained as yellow oil (200 mg, 67% yield).sup.1H NMR (400 MHz, CDCl.sub.3) : 7.30-7.29 (m, 1H), 7.00-6.99 (m, 1H), 2.56-2.41 (m, 4H), 2.03-1.95 (m, 1H), 1.82-1.75 (m, 1H).

Intermediate 76-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-ol

[0286] ##STR00036##

Step 1

[0287] Sodium borohydride (710 mg, 18.68 mmol) was added at 0 C. to a solution of 6-(trifluoromethyl)-3,4-dihydronaphthalen-1(2H)-one (2.0 g, 9.3 mmol) in MeOH (50 mL), and the mixture was allowed to stir at room temperature for 0.5 hr. The reaction mixture was concentrated under reduced pressure, quenched with water (50 mL) and extracted with EtOAc (260 mL). The combined organic layers were washed with brine and dried. The filtrate was concentrated under reduced pressure and the residue was purified by flash column chromatography on silica (25 g, 0-20% MTBE/petroleum ether) to give 6-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-ol (2.0 g, 99% yield) as white solid. LCMS m/z 199.0 (M-OH)+(ES+).

Step 2

[0288] To the solution of 6-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-ol (1.0 g, 4.6 mmol), (S)-1-(benzyloxycarbonyl)pyrrolidine-2-carboxylic acid (1.7 g, 6.9 mmol), and DMAP (847 mg, 6.9 mmol) in DCM (30 mL) was added EDCI (1.78 g, 9.3 mmol) and DIPEA (2.4 g, 18.5 mmol) at 0 C., and the resulting mixture was stirred at room temperature overnight. The reaction mixture was quenched with aq HCl (0.5 N) and the pH adjusted to 5. The organic layer was separated and the aqueous layer extracted with DCM (240 mL). The combined organic layers were washed with brine, dried and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (0-10% MTBE/petroleum ether) to give (S)-1-benzyl 2-(6-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-yl) pyrrolidine-1,2-dicarboxylate (1.9 g, 91% yield) as yellow oil. LCMS m/z 470.0 (M+Na)+ (ES+).

[0289] (S)-1-benzyl 2-(6-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-yl) pyrrolidine-1,2-dicarboxylate (1.90 g, 3.00 mmol) was separated by SFC (Column: CHIRALPAK AD-5 (30*250 mm 5 m) (Daicel); Column temperature: 35 C. CO.sub.2 flow Rate: 36 mL/min; co solvent flow rate: 9 mL/min; total flow rate: 45 mL/min. Co solvent: MeOH. Collection wavelength: 215 nm)). The SFC fractions were concentrated under reduced pressure to give (S)-1-benzyl 2-(6-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-yl) pyrrolidine-1,2-dicarboxylate ISOMER 1 (890 mg, 99 ee %, 46% yield) and (S)-1-benzyl 2-(6-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-yl) pyrrolidine-1,2-dicarboxylate ISOMER 2 (800 mg, 100 ee %, 42% yield). The absolute configuration was arbitrarily assigned in the synthetic scheme.

[0290] Chiral HPLC: (Column: CHIRALPAKAD-3(4.6*100 mm)); Flow Rate: 2 mL/min; Co_solvent: 15% MeOH; collection wavelength: 200-400 nm) ISOMER 1: Rt=1.764 min; ISOMER 2: Rt=2.137 min.

Step 3

[0291] A solution of (S)-1-benzyl 2-(6-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-yl) pyrrolidine-1,2-dicarboxylate (ISOMER 1, 890 mg, 2.0 mmol) and NaOH (159 mg, 4.0 mmol) in MeOH/H.sub.2O (5 mL/2.5 mL) was stirred at room temperature overnight. The mixture was concentrated under reduced pressure and pH was adjusted to 5 with addition of aq HCl (0.5 N). The organic layer was separated and the aqueous layer extracted with EtOAc (25 mL). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and filtered. The filtrate was concentrated under reduced pressure to give 6-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-ol ISOMER 1 (400 mg, 93% yield) as yellow solid. LCMS m/z 199.2 (MOH)+ (ES+).

Example 1(S)-3-((1-(4-(trifluoromethyl)phenyl)ethoxy)carbonyl)but-3-enoic acid

[0292] ##STR00037##

Step 1

[0293] A mixture of (S)-1-(4-(trifluoromethyl)phenyl)ethanol (190 mg, 1.0 mmol), 2-methylene-4-oxo-4-(2,2,2-trichloroethoxy)butanoic acid (260 mg, 1.0 mmol), DCC (309 mg, 1.5 mmol) and DMAP (24 mg, 0.2 mmol) in DCM (7 mL) was stirred at room temperature for 1 hr. The mixture was filtered, the filtrate concentrated under reduced pressure, and the residue purified by flash column chromatography on silica (0-10% MTBE/petroleum ether) to give (S)-4-(2,2,2-trichloroethyl) 1-(1-(4-(trifluoromethyl)phenyl)ethyl) 2-methylenesuccinate (270 mg, 62% yield) as a light yellow oil. LCMS m/z 454.8 (M+Na)+ (ES+).

Step 2

[0294] A mixture of (S)-4-(2,2,2-trichloroethyl) 1-(1-(4-(trifluoromethyl)phenyl)ethyl) 2-methylenesuccinate (270 mg, 0.6 mmol), zinc powder (201 mg, 3.1 mmol) and NH.sub.4OAc (477 mg, 6.2 mmol) in THF (2.5 mL) and H.sub.2O (0.8 mL) was stirred at room temperature for 4 hrs. The reaction mixture was filtered and the filtrate was acidified to pH 4-5 with aq HCl (0.5 N) and extracted with tert-butyl methyl ether (25 mL). The combined organic phases were washed with brine, dried (magnesium sulfate) and concentrated under reduced pressure. The residue was purified by prep-HPLC (Column: Waters SUNFIRE Prep C18 OBD 10 m 19250 mm; Flow Rate: 20 mL/min; solvent system: MeCN/(0.2% trifluoroacetic acid/water) MeCN gradient: 50-95%; collection wavelength: 214 nm). The fractions were concentrated under reduced pressure to remove MeCN, and the residue was lyophilized to give (S)-3-((1-(4-(trifluoromethyl)phenyl)ethoxy)carbonyl)but-3-enoic acid (58.1 mg, 30% yield) as yellow oil. LCMS m/z 324.9 (M+Na)+ (ES+).

[0295] .sup.1H NMR (400 MHz, DMSO-d.sub.6) : 12.40 (br, 1H), 7.73 (d, J=8.4 Hz, 2H), 7.61 (d, J=8.4 Hz, 2H), 6.26 (d, J=0.8 Hz, 1H), 5.93 (q, J=6.8 Hz, 1H), 5.83 (s, 1H), 3.31 (s, 2H), 1.50 (d, J=6.4 Hz, 3H).

Example 23-((1-(4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoic acid

[0296] ##STR00038##

[0297] Prepared from Intermediate 1 and Intermediate 2 using a similar procedure to (S)-3-((1-(4-(trifluoromethyl)phenyl)ethoxy)carbonyl)but-3-enoic acid.

Step 1

[0298] 4-(2,2,2-trichloroethyl) 1-(1-(4-(trifluoromethyl)phenyl)cyclobutyl) 2-methylenesuccinate (260 mg, 61% yield) was obtained as a colorless oil. LCMS m/z 480.8 (M+Na)+ (ES+).

Step 2

[0299] 3-((1-(4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoic acid (82.5 mg, 45% yield) was obtained as white solid. LCMS m/z 351.0 (M+Na)+ (ES+). .sup.1H NMR (400 MHz, DMSO-d6) : 12.39 (s, 1H), 7.69 (d, J=8.4 Hz, 2H), 7.63 (d, J=8.4 Hz, 2H), 6.15 (s, 1H), 5.75 (s, 1H), 3.25 (s, 2H), 2.61-2.51 (m, 4H), 1.99-1.92 (m, 1H), 1.79-1.72 (m, 1H).

Example 33-((1-(3-fluoro-4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoic acid

[0300] ##STR00039##

[0301] Prepared from Intermediate 1 and Intermediate 3 using a similar procedure to (S)-3-((1-(4-(trifluoromethyl)phenyl)ethoxy)carbonyl)but-3-enoic acid.

Step 1

[0302] 1-(1-(3-fluoro-4-(trifluoromethyl)phenyl)cyclobutyl) 4-(2,2,2-trichloroethyl) 2-methylenesuccinate (490 mg, 52% yield) was obtained as a colorless oil. LCMS m/z 498.8 (M+Na)+ (ES+).

Step 2

[0303] 3-((1-(3-fluoro-4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoic acid (134.2 mg, 38% yield) was obtained as white solid. LCMS m/z 368.8 (M+Na)+ (ES+).

[0304] .sup.1H NMR (400 MHz, DMSO-d6) : 12.42 (s, 1H), 7.76 (t, J=8.0 Hz, 1H), 7.53 (d, J=12.0 Hz, 1H), 7.48 (d, J=8.0 Hz, 1H), 6.20 (s, 1H), 5.79 (s, 1H), 3.30 (s, 2H), 2.64-2.53 (m, 4H), 2.00-1.94 (m, 1H), 1.85-1.78 (m, 1H).

Example 43-(((3-(4-(trifluoromethyl)phenyl)oxetan-3-yl)oxy)carbonyl)but-3-enoic acid

[0305] ##STR00040##

[0306] Prepared from Intermediate 1 and Intermediate 4 using a similar procedure to (S)-3-((1-(4-(trifluoromethyl)phenyl)ethoxy)carbonyl)but-3-enoic acid.

Step 1

[0307] 4-(2,2,2-trichloroethyl) 1-(3-(4-(trifluoromethyl)phenyl)oxetan-3-yl) 2-methylenesuccinate (440 mg, 52% yield) was obtained as a colorless oil. LCMS m/z 460.8 (M+H)+(ES+).

Step 2

[0308] 3-(((3-(4-(trifluoromethyl)phenyl)oxetan-3-yl)oxy)carbonyl)but-3-enoic acid (86.2 mg, 30% yield) was obtained as white solid. LCMS m/z 331.0 (M+H)+(ES+). .sup.1H NMR (400 MHz, DMSO-d6) : 12.50 (br, 1H), 7.79 (d, J=8.4 Hz, 2H), 7.74 (d, J=8.4 Hz, 2H), 6.34 (s, 1H), 5.92 (s, 1H), 4.96 (d, J=8.0 Hz, 2H), 4.87 (d, J=8.0 Hz, 2H), 3.36 (s, 2H).

Example 53-((1-(5-(trifluoromethyl)pyridin-2-yl)cyclobutoxy)carbonyl)but-3-enoic acid

[0309] ##STR00041##

[0310] Prepared from Intermediate 1 and Intermediate 5 using a similar procedure to (S)-3-((1-(4-(trifluoromethyl)phenyl)ethoxy)carbonyl)but-3-enoic acid.

Step 1

[0311] 4-(2,2,2-trichloroethyl) 1-(1-(5-(trifluoromethyl)pyridin-2-yl)cyclobutyl) 2-methylenesuccinate (360 mg, 42% yield) was obtained as a colorless oil. LCMS m/z 460.0 (M+H)+ (ES+).

Step 2

[0312] 3-((1-(5-(trifluoromethyl)pyridin-2-yl)cyclobutoxy)carbonyl)but-3-enoic acid (72.2 mg, 28% yield) was obtained as white solid. LCMS m/z 330.0 (M+H)+ (ES+). .sup.1H NMR (400 MHz, DMSO-d6) : 12.43 (s, 1H), 8.99 (t, J=1.2 Hz, 1H), 8.16 (dd, J=8.4, 2.0 Hz, 1H), 7.55 (d, J=8.4 Hz, 1H), 6.25 d, J=0.8 Hz, 1H), 5.84 (d, J=0.8 Hz, 1H), 3.33 (s, 2H), 2.75-2.69 (m, 2H), 2.55-2.47 (m, 2H), 2.01-1.92 (m, 2H).

Example 63-((1-(5-(trifluoromethyl)thiophen-2-yl)cyclobutoxy)carbonyl)but-3-enoic acid

[0313] ##STR00042##

[0314] Prepared from Intermediate 1 and Intermediate 6 using a similar procedure to (S)-3-((1-(4-(trifluoromethyl)phenyl)ethoxy)carbonyl)but-3-enoic acid.

Step 1

[0315] 1-(1-(5-(trifluoromethyl)thiophen-2-yl)cyclobutyl) 2-methylenesuccinate (280 mg, 67% yield) was obtained as a colorless oil. LCMS m/z 486.7 (M+H)+ (ES+).

Step 2

[0316] 3-((1-(5-(trifluoromethyl)thiophen-2-yl)cyclobutoxy)carbonyl)but-3-enoic acid (86.0 mg, 43% yield) was obtained as white solid. LCMS m/z 357.1 (M+Na)+ (ES+). .sup.1H NMR (400 MHz, DMSO-d6) : 12.39 (s, 1H), 7.60 (dd, J=4.0, 1.2 Hz, 1H), 7.32 (d, J=4.0 Hz, 1H), 6.17 (d, J=1.2 Hz, 1H), 5.80 (d, J=0.8 Hz, 1H), 3.27 (s, 2H), 2.67-2.57 (m, 4H), 1.97-1.97 (m, 1H), 1.84-1.77 (m, 3H).

Example 73-(((6-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-yl)oxy)carbonyl)but-3-enoic acid

[0317] ##STR00043##

[0318] Prepared from Intermediate 1 and Intermediate 7 using a similar procedure to (S)-3-((1-(4-(trifluoromethyl)phenyl)ethoxy)carbonyl)but-3-enoic acid.

Step 1

[0319] 4-(2,2,2-trichloroethyl) 1-(6-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-yl) 2-methylenesuccinate (300 mg, 71% yield) was obtained as a colorless oil. LCMS m/z 481.0 (M+Na)+ (ES+).

Step 2

[0320] 3-(((6-(trifluoromethyl)-1,2,3,4-tetrahydronaphthalen-1-yl)oxy)carbonyl)but-3-enoic acid (102.4 mg, 48% yield) was obtained as white solid. The absolute configuration was arbitrarily assigned. LCMS m/z 351.1 (M+Na)+ (ES+). .sup.1H NMR (400 MHz, DMSO-d6) : 12.39 (br, 1H), 7.54-7.50 (m, 2H), 7.43 (d, J=8.0 Hz, 1H), 6.16 (d, J=1.2 Hz, 1H), 5.99 (t, J=4.4 Hz, 1H), 5.78 (d, J=0.8 Hz, 1H), 3.28 (s, 2H), 2.95-2.88 (m, 1H), 2.83-2.75 (m, 1H), 2.05-1.98 (m, 1H), 1.92-1.78 (m, 1H).

Example 84-methyl 1-(1-(4-(trifluoromethyl)phenyl)cyclobutyl) 2-methylenesuccinate

[0321] ##STR00044##

[0322] DCC (408 mg, 2.0 mmol) was added to a mixture of 1-(4-(trifluoromethyl)phenyl)cyclobutan-1-ol (Intermediate 2, 0.3 g, 1.3 mmol), 4-methoxy-2-methylene-4-oxobutanoic acid (200 mg, 1.3 mmol) and DMAP (16 mg, 0.1 mmol) in DCM (4 mL) at 0 C. The mixture was stirred at RT for 2 hrs, filtered, and the filtrate concentrated under reduced pressure. The crude product was purified by chromatography on silica (0-25% MTBE/heptane) to afford 4-methyl 1-(1-(4-(trifluoromethyl)phenyl)cyclobutyl) 2-methylenesuccinate (241 mg, 0.7 mmol) as a colourless oil. .sup.1H NMR (400 MHz, DMSO-d6) 7.74 (d, J=8.3 Hz, 2H), 7.64 (d, J=8.2 Hz, 2H), 6.25-6.20 (m, 1H), 5.86-5.81 (m, 1H), 3.59 (s, 3H), 3.38 (s, 2H), 2.66-2.52 (m, 4H), 2.05-1.91 (m, 1H), 1.85-1.71 (m, 1H).

Example 91-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(methylamino)-2-methylene-4-oxobutanoate

[0323] ##STR00045##

[0324] A solution of T3P (50 wt % in EtOAc, 0.8 mL, 1.4 mmol) was added dropwise to a stirred solution of 3-((1-(4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoic acid (Example 2, 250 mg, 0.7 mmol), methylamine (2 M in THF, 0.5 mL, 1.0 mmol) and triethylamine (0.2 mL, 1.4 mmol) in EtOAc (3 mL) at RT. The mixture was stirred at RT for 1 hr then diluted with brine (20 mL) and extracted with EtOAc (220 mL). The combined organic phases were dried (magnesium sulfate) and concentrated under reduced pressure. The residue was purified by chromatography on Reverse Phase Flash C18 (5-75% MeCN/(0.1% Formic acid in Water)) to afford 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(methylamino)-2-methylene-4-oxobutanoate (183 mg, 0.5 mmol) as a white solid. LCMS m/z 364.3 (M+Na).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, DMSO-d6) 7.84-7.77 (m, 1H), 7.72 (d, J=8.3 Hz, 2H), 7.64 (d, J=8.3 Hz, 2H), 6.20-6.10 (m, 1H), 5.75-5.66 (m, 1H), 3.11 (s, 2H), 2.63-2.53 (m, 7H), 2.06-1.90 (m, 1H), 1.86-1.69 (m, 1H).

Example 101-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(dimethylamino)-2-methylene-4-oxobutanoate

[0325] ##STR00046##

[0326] Prepared by an analogous method to Example 10 using dimethylamine (2 M in THF). Obtained as a colourless oil (202 mg, 54%). LCMS m/z 378.3 (M+Na).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, DMSO) 7.72 (d, J=8.4 Hz, 2H), 7.65 (d, J=8.3 Hz, 2H), 6.15-6.09 (m, 1H), 5.65-5.59 (m, 1H), 3.36 (s, 2H), 2.98 (s, 3H), 2.82 (s, 3H), 2.62-2.52 (m, 4H), 2.04-1.90 (m, 1H), 1.86-1.71 (m, 1H).

Example 111-(4-(trifluoromethyl)phenyl)cyclobutyl 4-amino-2-methylene-4-oxobutanoate

[0327] ##STR00047##

[0328] Prepared by an analogous method to Example 10 starting from ammonium chloride Obtained as a pale yellow solid (21 mg, 59% yield) LCMS m/z 350.3 (M+Na).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, DMSO) 7.71 (d, J=8.4 Hz, 2H), 7.66 (d, J=8.4 Hz, 2H), 7.35 (s, 1H), 6.88 (s, 1H), 6.17-6.10 (m, 1H), 5.72-5.66 (m, 1H), 3.12 (s, 2H), 2.58 (t, J=7.9 Hz, 4H), 2.06-1.89 (m, 1H), 1.87-1.69 (m, 1H).

Example 121-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((2-(dimethylamino)ethyl)amino)-2-methylene-4-oxobutanoate

[0329] ##STR00048##

[0330] A mixture of 3-((1-(4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoic acid (200 mg, 0.61 mmol), N.sup.1,N.sup.1-dimethylethane-1,2-diamine (54 mg, 0.61 mmol), HATU (348 mg, 0.915 mmol) and triethylamine (132 mg, 132 mmol) in DMF (3 mL) was stirred at 0 C. for 2 hrs. The reaction mixture was quenched with water (4 mL), and extracted with ethyl acetate (35 mL). The organic layer was washed by brine, dried over Na.sub.2SO.sub.4, and concentrated under reduced pressure. The residue was purified by prep-HPLC (Column: Waters X-bridge Prep C18 OBD 10 m 19250 mm; Flow Rate: 20 mL/min; solvent system: MeCN/(10 mmol/L NH.sub.4HCO.sub.3/water) gradient: MeCN: 40-95%; collection wavelength: 214 nm). The fractions were concentrated under reduced pressure to remove MeCN, and lyophilized to give 1-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(2-(dimethylamino)ethylamino)-2-methylene-4-oxobutanoate (78 mg, 32% yield) as white solid. LCMS m/z 399.3 (M+H).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, CDCl3) : 7.63-7.54 (m, 4H), 6.41 (br s, 1H), 6.37 (d, J=0.8 Hz, 1H), 5.82 (d, J=1.2 Hz, 1H), 3.30 (q, J=5.6 Hz, 2H), 3.18 (s, 2H), 2.68 (dd, J=8.6, 6.9 Hz, 4H), 2.38 (t, J=6 Hz, 2H), 2.21 (s, 6H), 2.07-1.99 (m, 1H), 1.86-1.77 (m, 1H).

Example 131-(4-(trifluoromethyl)phenyl)cyclobutyl 2-methylene-4-((2-morpholinoethyl)amino)-4-oxobutanoate

[0331] ##STR00049##

[0332] Prepared by an analogous method to Example 12 starting from 2-morpholinoethanamine. Obtained as a white solid (143 mg, 53% yield). LCMS m/z 441.3 (M+H).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, DMSO-d6) : 7.85 (t, J=5.6 Hz, 1H), 7.72 (d, J=8.4 Hz, 2H), 7.65 (d, J=8.4 Hz, 2H), 6.16 (d, J=1.2 Hz, 1H), 5.73 (d, J=0.8 Hz, 1H), 3.54 (t, J=4.8 Hz, 4H), 3.17 (q, J=6 Hz, 2H), 3.13 (s, 2H), 2.62-2.55 (m, 4H), 2.33-2.29 (m, 6H), 2.00-1.94 (m, 1H), 1.83-1.75 (m, 1H).

Example 141-(4-(trifluoromethyl)phenyl)cyclobutyl 2-methylene-4-((1-methylpiperidin-4-yl)amino)-4-oxobutanoate

[0333] ##STR00050##

[0334] Prepared by an analogous method to Example 12 starting from 1-methylpiperidin-4-amine. Obtained as a white solid (94 mg, 36% yield). LCMS m/z 425.4 (M+H).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, DMSO-d6) : 7.83 (d, J=7.6 Hz, 1H), 7.72 (d, J=8 Hz, 2H), 7.64 (d, J=8.4 Hz, 2H), 6.13 (d, J=1.2 Hz, 1H), 5.68 (d, J=1.2 Hz, 1H), 3.52-3.43 (m, 1H), 3.12 (s, 2H), 2.69-2.60 (m, 2H), 2.60-2.57 (m, 4H), 2.12 (s, 3H), 2.00-1.87 (m, 3H), 1.82-1.75 (m, 1H), 1.69-1.60 (m, 2H), 1.42-1.32 (m, 2H).

Example 151-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(((1H-imidazol-2-yl)methyl)amino)-2-methylene-4-oxobutanoate

[0335] ##STR00051##

[0336] Prepared by an analogous method to Example 12 starting from 1H-imidazol-2-yl)methanamine. Obtained as a white solid (40 mg, 16% yield). LCMS m/z 408.2 (M+H).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, DMSO-d6) : 11.74 (br s, 1H), 8.40 (t, J=5.2 Hz, 1H), 7.71 (d, J=8.4 Hz, 2H), 7.63 (d, J=8.4 Hz, 2H), 6.99 (s, 1H), 6.80 (s, 1H), 6.16 (d, J=1.2 Hz, 1H), 5.73 (d, J=1.2 Hz, 1H), 4.27 (d, J=5.6 Hz, 2H), 3.20 (s, 2H), 2.56-2.51 (m, 4H), 1.97-1.92 (m, 1H), 1.79-1.74 (m, 1H).

Example 161-(4-(trifluoromethyl)phenyl)cyclobutyl 4-(((1-methyl-1H-imidazol-2-yl)methyl)amino)-2-methylene-4-oxobutanoate

[0337] ##STR00052##

[0338] Prepared by an analogous method to Example 12 starting from 1-methyl-1H-imidazol-2-yl)methanamine. Obtained as a white solid (174 mg, 68% yield). LCMS m/z 422.2 (M+H).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, DMSO-d6) : 8.40 (t, J=5.2 Hz, 1H), 7.72 (d, J=8.4 Hz, 2H), 7.63 (d, J=8.4 Hz, 2H), 7.05 (d, J=1.2 Hz, 1H), 6.77 (d, J=1.2 Hz s, 1H), 6.16 (d, J=1.2 Hz, 1H), 5.73 (d, J=0.8 Hz, 1H), 4.32 (d, J=5.2 Hz, 2H), 3.53 (s, 3H), 3.18 (s, 2H), 2.60-2.54 (m, 4H), 2.01-1.91 (m, 1H), 1.82-1.71 (m, 1H).

Example 171-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((1-amino-3-hydroxy-1-oxopropan-2-yl)amino)-2-methylene-4-oxobutanoate

[0339] ##STR00053##

[0340] Prepared by an analogous method to Example 12 starting from 2-amino-3-hydroxypropanamide. Obtained as a white solid (127 mg, 40% yield). LCMS m/z 437.6 (M+Na).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, DMSO-d6) : 7.91 (d, J=8.1 Hz, 1H), 7.72 (d, J=8.3 Hz, 2H), 7.64 (d, J=8.2 Hz, 2H), 7.26 (s, 1H), 7.09 (s, 1H), 6.17-6.12 (m, 1H), 5.74-5.70 (m, 1H), 4.86 (t, J=5.5 Hz, 1H), 4.28-4.19 (m, 1H), 3.63-3.48 (m, 2H), 3.30-3.18 (m, 2H), 2.58 (t, J=8.7 Hz, 4H), 2.06-1.89 (m, 1H), 1.84-1.70 (m, 1H).

Example 181-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((1,1-dioxidothietan-3-yl)amino)-2-methylene-4-oxobutanoate

[0341] ##STR00054##

[0342] Prepared by an analogous method to Example 9 starting from 3-aminoheptane 1,1-dioxide. Obtained as a white solid (193 mg, 65% yield). LCMS m/z 454.2 (M+Na).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, DMSO-d6) : 8.75 (d, J=5.3 Hz, 1H), 7.73 (d, J=8.3 Hz, 2H), 7.64 (d, J=8.2 Hz, 2H), 6.19 (d, J=1.4 Hz, 1H), 5.75 (d, J=1.4 Hz, 1H), 4.57-4.48 (m, 2H), 4.38-4.30 (m, 1H), 4.03-3.95 (m, 2H), 3.19 (s, 2H), 2.64-2.52 (m, 4H), 2.02-1.92 (m, 1H), 1.85-1.73 (m, 1H).

Example 191-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((2-hydroxyethyl)amino)-2-methylene-4-oxobutanoate

[0343] ##STR00055##

[0344] Prepared by an analogous method to Example 9 starting from 2-aminoethane-1-ol. Obtained as a white solid (145 mg, 54% yield). LCMS m/z 394.2 (M+Na).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, DMSO-d6) : 7.91 (t, J=5.7 Hz, 1H), 7.72 (d, J=8.3 Hz, 2H), 7.65 (d, J=8.2 Hz, 2H), 6.17-6.10 (m, 1H), 5.73-5.66 (m, 1H), 4.65 (t, J=5.4 Hz, 1H), 3.39 (q, J=6.0 Hz, 2H), 3.18-3.09 (m, 4H), 2.62-2.53 (m, 4H), 2.04-1.90 (m, 1H), 1.85-1.71 (m, 1H).

Example 201-(4-(trifluoromethyl)phenyl)cyclobutyl 2-methylene-4-oxo-4-((tetrahydro-2H-pyran-4-yl)amino)butanoate

[0345] ##STR00056##

[0346] Prepared by an analogous method to Example 9 starting from tetrahydro-2H-pyran-4-amine. Obtained as a white solid (219 mg, 74% yield). LCMS m/z 434.0 (M+Na).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, DMSO-d6) : 7.91 (d, J=7.6 Hz, 1H), 7.72 (d, J=8.3 Hz, 2H), 7.64 (d, J=8.3 Hz, 2H), 6.19-6.08 (m, 1H), 5.74-5.62 (m, 1H), 3.86-3.67 (m, 3H), 3.31-3.29 (m, 2H), 3.13 (s, 2H), 2.64-2.53 (m, 4H), 2.06-1.90 (m, 1H), 1.87-1.72 (m, 1H), 1.70-1.59 (m, 2H), 1.45-1.28 (m, 2H).

Example 211-(4-(trifluoromethyl)phenyl)cyclobutyl 4-((2-amino-2-oxoethyl)amino)-2-methylene-4-oxobutanoate

[0347] ##STR00057##

[0348] Prepared by an analogous method to Example 9 starting from 2-aminoacetamide. Obtained as a white solid (35 mg, 13% yield). LCMS m/z 385.0 (M+H).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, DMSO-d6) : 8.10 (t, J=5.8 Hz, 1H), 7.72 (d, J=8.3 Hz, 2H), 7.65 (d, J=8.3 Hz, 2H), 7.24 (s, 1H), 7.04 (s, 1H), 6.20-6.14 (m, 1H), 5.78-5.69 (m, 1H), 3.64 (d, J=5.7 Hz, 2H), 3.21 (s, 2H), 2.58 (t, J=7.9 Hz, 4H), 2.04-1.89 (m, 1H), 1.83-1.69 (m, 1H).

Example 222-((3-((1-(4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoyl)oxy)acetic acid

[0349] ##STR00058##

Step 1

[0350] To a solution of ethyl 2,2,2-trichloroethan-1-ol (8.9 g, 60 mmol) and pyridine (6 g, 75 mmol) in THF (150 mL) was added 2-bromoacetyl bromide (10 g, 50 mmol) at 0 C., and the reaction mixture was stirred at room temperature for 4 hrs. The reaction mixture was quenched with water (400 mL) and extracted with MTBE (3100 mL). The combined organic layers were washed with aq HCl aq. (0.5N, 250 mL) and brine, dried (magnesium sulfate) and concentrated under reduced pressure. The residue was purified by column chromatography on silica (eluted by 0-5% MTBE/petroleum ether) to give 2,2,2-trichloroethyl 2-bromoacetate (9.5 g, 71% yield) as light yellow oil. .sup.1H NMR (400 MHz, CDC.sub.3) : 4.82 (s, 2H), 3.98 (s, 2H).

Step 2

[0351] A mixture of 3-((1-(4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoic acid (300 mg, 0.9 mmol), 2,2,2-trichloroethyl 2-bromoacetate (245 mg, 0.915 mmol) and K.sub.2CO.sub.3 (152 mg, 1.1 mmol) in acetone (3 mL) was stirred at room temperature overnight. The reaction mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by flash column chromatography on silica (0-10% MTBE/petroleum ether) to give 4-(2-oxo-2-(2,2,2-trichloroethoxy)ethyl) 1-(1-(4-(trifluoromethyl)phenyl)cyclobutyl) 2-methylenesuccinate (200 mg, 42% yield) as a yellow oil. LCMS m/z 539.7 (M+Na).sup.+ (ES.sup.+).

Step 3

[0352] A mixture of 4-(2-oxo-2-(2,2,2-trichloroethoxy)ethyl) 1-(1-(4-(trifluoromethyl)phenyl)cyclobutyl) 2-methylenesuccinate (200 mg, 0.4 mmol), zinc powder (127 mg, 2.0 mmol) and NH.sub.4OAc (300 mg, 3.9 mmol) in THF (1.5 mL) and H.sub.2O (0.5 mL) was stirred at room temperature for 2 hrs. The reaction mixture was filtered and the filtrate was acidified with aq HCl (0.5 N). until pH 5, and extracted with MTBE (25 mL). The combined organic phases were washed with brine, dried (magnesium sulfate) and concentrated under reduced pressure. The residue was purified by prep-HPLC (Column: Waters SUNFIRE Prep C18 OBD 10 m 19250 mm; Flow Rate: 20 mL/min; solvent system: MeCN/(0.2% formic acid/water) MeCN gradient: 50-95%; collection wavelength: 214 nm). The fractions were concentrated under reduced pressure to remove MeCN, and the residue was lyophilized to give 2-(3-((1-(4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoyloxy)acetic acid (55.43 mg, 36% yield) as light yellow oil. LCMS m/z 408.9 (M+Na).sup.+ (ES.sup.+). .sup.1H NMR (400 MHz, DMSO-d6) : 13.099 (br, 1H), 7.73 (d, J=8.4 Hz, 2H), 7.65 (d, J=8.4 Hz, 2H), 6.24 (d, J=1.2 Hz, 1H), 5.89 (d, J=0.8 Hz, 1H), 4.58 (s, 2H), 3.48 (s, 2H), 2.58 (t, J=8.8 Hz, 4H), 1.98-1.95 (m, 1H), 1.80-1.75 (m, 1H).

Example 233-methyl-4-oxo-4-(1-(4-(trifluoromethyl)phenyl)cyclobutoxy)but-2-enoic acid

[0353] ##STR00059##

[0354] A mixture of 3-((1-(4-(trifluoromethyl)phenyl)cyclobutoxy)carbonyl)but-3-enoic acid (Example 2, 150 mg, 0.46 mmol) in THF (2 mL) and triethylamine (1 mL) was stirred at 65 C. for 2 days. The solvent was removed under reduced pressure; the residue was diluted with MTBE (3 mL) and acidified with aq HCl (0.5 N) until pH 4-5. separated and extracted with tert-butyl methyl ether (2 203 mL). The combined organic phases were washed with brine, dried (magnesium sulfate) and concentrated under reduced pressure. The residue was purified by prep-HPLC (Column: Waters X-Bridge C18 OBD 10 m 19250 mm; Flow Rate: 20 mL/min; solvent system: MeCN/(0.2% formic acid/water) MeCN gradient:: 50-95%; collection wavelength: 214 nm). The fractions were concentrated under reduced pressure to remove MeCN, and the residue was lyophilized to give 3-methyl-4-oxo-4-(1-(4-(trifluoromethyl)phenyl)cyclobutoxy)but-2-enoic acid (45.2 mg, 31% yield) as white solid. .sup.1H NMR (400 MHz, DMSO-d6) : 13.02 (br, 1H), 7.75 (d, J=8.4 Hz, 2H), 7.69 (d, J=8.4 Hz, 2H), 6.70 (m, 1H), 2.65-2.62 (m, 4H), 2.12 (d, J=1.2 Hz, 3H), 2.02-1.91 (m, 1H), 1.82-1.70 (m, 1H).

Biological Example 1THP-1 AlphaLISA IL-1 and IL-6 Cytokine Assay

Measuring Inhibitory Effects on IL-1, and IL-6 Cytokine Output from THP-1s

[0355] The cytokine inhibition profiles of compounds of formula (I) were determined in a differentiated THP-1 cell assay. All assays were performed in RPMI-1640 growth medium (Gibco), supplemented with 10% fetal bovine serum (FBS; Gibco), 1% penicillin-streptomycin and 1% sodium pyruvate unless specified otherwise. The IL-1 and IL-6 cytokine inhibition assays were each run in a background of differentiated THP-1 cells as described below. All reagents described were from Sigma-Aldrich unless specified otherwise. Compounds were prepared as 10 mM DMSO stocks.

Assay Procedure

[0356] THP-1 cells were expanded as a suspension up to 80% confluence in appropriate growth medium. Cells were harvested, suspended, and treated with an appropriate concentration of phorbol 12-myristate 13-acetate (PMA) over a 72 hr period (37 C./5% CO.sub.2).

[0357] Following 72 hrs of THP-1 cell incubation, cellular medium was removed and replaced with fresh growth media containing 1% of FBS. Working concentrations of compounds were prepared separately in 10% FBS treated growth medium and pre-incubated with the cells for 30 minutes (37 C./5% CO.sub.2). Following the 30 minute compound pre-incubation, THP-1s were treated with an appropriate concentration of LPS and the THP-1s were subsequently incubated for a 24 hr period (37 C./5% CO.sub.2). An appropriate final concentration of Nigericin was then dispensed into the THP-1 plates and incubated for 1 hour (37 C./5% CO.sub.2) before THP-1 supernatants were harvested and collected in separate polypropylene 96-well holding plates.

[0358] Reagents from each of the IL-1 and IL-6 commercial kits (Perkin Elmer) were prepared and run according to the manufacturer's instructions. Subsequently, fluorescence signal detection in a microplate reader was measured (EnVision Multilabel Reader, Perkin Elmer).

[0359] Percentage inhibition was calculated per cytokine by normalizing the sample data to the high and low controls used within each plate (+/LPS respectively). Percentage inhibition was then plotted against compound concentration and the 50% inhibitory concentration (IC.sub.50) was determined from the resultant concentration-response curve.

[0360] The data for all compounds of formula (I) tested in this assay are presented in Table 1 below. Dimethyl itaconate and 1-monomethyl itaconate were included as comparator compounds.

TABLE-US-00001 TABLE 1 THP-1 cell IL-1 and IL-6 IC.sub.50 values (M) Compound IL-1 (IC.sub.50) IL-6 (IC.sub.50) dimethyl itaconate >100 >100 1-monomethyl itaconate >100 >100 Example 1 5.4 54.8 Example 2 4.4 >100 Example 3 4.8 53.7 Example 4 9.2 >100 Example 5 9.0 >31.6 Example 6 9.5 >100 Example 7 7.9 65.6 Example 8 7.6 10.8 Example 9 12.6 11.1 Example 10 60.0 13.8 Example 11 9.9 13.2 Example 12 3.1 4.0 Example 13 6.5 8.1 8.5* Example 14 5.2 3.9 Example 15 5.3 7.3 Example 16 5.9 5.3 Example 17 15.3 12.4 Example 18 5.2 3.8 Example 19 2.5 3.8 Example 20 2.9 2.6 Example 21 7.2 3.9 Example 22 15.2 >31.6 Example 23 4.0 >100 *repeated value

[0361] These results reveal that compounds of formula (I) are expected to have anti-inflammatory activity as shown by their IC.sub.50 values for inhibition of IL-1 and IL-6 release in this assay. All compounds of the invention tested exhibited improved IL-1 lowering properties (IC.sub.50 values) compared with dimethyl itaconate and 1-monomethyl itaconate. Preferred compounds of the invention tested exhibited improved IL-1 and IL-6 lowering properties (IC.sub.50 values) compared with dimethyl itaconate and 1-monomethyl itaconate

Biological Example 2NRF2 Activation Assay

Measuring Compound Activation Effects on the Anti-Inflammatory Transcription Factor NRF2 in DiscoverX PathHunter NRF2 Translocation Kit

[0362] Potency and efficacy of compounds of formula (I) against the target of interest to activate NRF2 (nuclear factor erythroid 2-related factor 2) were determined using the PathHunter NRF2 translocation kit (DiscoverX). The NRF2 translocation assay was run using an engineered recombinant cell line, utilising enzyme fragment complementation to determine activation of the Keap1-NRF2 protein complex and subsequent translocation of NRF2 into the nucleus. Enzyme activity was quantified using a chemiluminescent substrate consumed following the formation of a functional enzyme upon PK-tagged NRF2 translocation into the nucleus.

[0363] Additionally, a defined concentration of dimethyl fumarate was used as the High control to normalise test compound activation responses.

Assay Procedure

[0364] U2OS PathHunter eXpress cells were thawed from frozen prior to plating. Following plating, U2OS cells were incubated for 24 hrs (37 C./5% CO.sub.2) in commercial kit provided cell medium.

[0365] Following 24 hrs of U2OS incubation, cells were directly treated with an appropriate final concentration of compound.

[0366] Following compound treatment, the U2OS plates were incubated for a further 6 hours (37 C./5% CO.sub.2) before detection reagent from the PathHunter NRF2 commercial kit was prepared and added to test plates according to the manufacturer's instructions. Subsequently, the luminescence signal detection in a microplate reader was measured (PHERAstar, BMG Labtech).

[0367] Percentage activation was calculated by normalising the sample data to the high and low controls used within each plate (+/DMF). Percentage activation/response was then plotted against compound concentration and the 50% activation concentration (EC.sub.50) was determined from the plotted concentration-response curve.

[0368] The data for all compounds of formula (I) tested in this assay are presented in Table 2 below.

[0369] Dimethyl itaconate and 1-monomethyl itaconate were included as comparator compounds.

TABLE-US-00002 TABLE 2 NRF2 activation GSH +GSH Compound EC.sub.50 (M) E.sub.max (%) EC.sub.50 (M) E.sub.max (%) dimethyl itaconate 21.7 126 56.4 77 1-monomethyl 30.1 44 NT* NT itaconate Example 2 >100 26 >100 14 Example 8 51 51 >100 38 Example 11 33.1 117 45.7 100 Example 13 38.8 174 48.0 84 Example 14 14.6 80 49.4 95 Example 15 26.3 131 51.3 86 Example 18 14.7 90 39.4 70 Example 19 40.2 210 50.7 102 Example 20 14.4 101 31.4 63 Example 21 38.0 184 55.0 86 Example 22 40.0 100 41.1 45 Example 23 2.8 39 >100 4 NT = not tested in this assay

[0370] For the most part, the compounds in Table 2 display relatively low NRF2 activating effects compared with the controls, as demonstrated by their EC.sub.50 and/or E.sub.max values for NRF2 activation, indicating that the ULP-lowering effect may not be solely a consequence of NRF2 activation. Thus, these compounds are expected to display reduced effects resulting from NRF2 activation.

Biological Example 3Hepatocyte Stability Assay

[0371] Defrosted cryo-preserved hepatocytes (viability >70%) were used to determine the metabolic stability of a compound via calculation of intrinsic clearance (CI.sub.int; a measure of the removal of a compound from the liver in the absence of blood flow and cell binding). Clearance data are particularly important for in vitro work as they can be used in combination with in vivo data to predict the half-life and oral bioavailability of a drug.

[0372] The metabolic stability in hepatocytes assay involved a time-dependent reaction using both positive and negative controls. The cells must be pre-incubated at 37 C. then spiked with test compound (and positive control); samples taken at pre-determined time intervals were analysed to monitor the change in concentration of the initial drug compound over 60 minutes. A buffer incubation reaction (with no hepatocytes present) acted as a negative control and two cocktail solutions, containing compounds with known high and low clearance values (verapamil/7-hydroxycoumarin and propranolol/diltiazem), acted as positive controls. [0373] 1. The assay was run with a cell concentration of 0.510.sup.6 cells/mL in Leibovitz buffer. [0374] 2. All compounds and controls were run in duplicate. [0375] 3. Compound concentration was 10 M. [0376] 4. All compounds and controls were incubated with both cells and buffer to show turnover is due to hepatic metabolism. [0377] 5. All wells on the incubation plate had 326.7 L of either cells or buffer added. [0378] 6. Prior to assay, cell and buffer-only incubation plates were preincubated for 10 mins at 37 C. [0379] 7. The assay was initiated by adding compounds, 3.3 L of 1 mM in 10% DMSO-90% Buffer; final DMSO concentration is 0.1%. [0380] 8. Samples were taken at regular timepoints (0, 5, 10, 20, 40, 60 min) until 60 mins. [0381] 9. Sample volume was 40 L and added to 160 L of crash solvent (acetonitrile with internal standard) and stored on ice. [0382] 10. At the end of the assay, the crash plates were centrifuged at 3500 rpm for 20 mins at 4 C. [0383] 11. 80 L of clear supernatant was removed and mixed with 80 L of deionised water before being analysed by LC-MS/MS.

[0384] Raw LC-MS/MS data were exported to, and analysed in, Microsoft Excel for determination of intrinsic clearance. The percentage remaining of a compound was monitored using the peak area of the initial concentration as 100%. Intrinsic clearance and half-life values were calculated using a graph of the natural log of percentage remaining versus the time of reaction in minutes. Half-life (min) and intrinsic clearance (CI.sub.int in L min.sup.1 10.sup.6 cells) values were calculated using the gradient of the graph (the elimination rate constant, k) and Equations 1 and 2.

[00001]$\begin{array}{cc}{t}_{\frac{1}{2}}=\frac{\ln 2}{k}& \left\{\mathrm{Equation}1\right\}\end{array}$$\begin{array}{cc}{\mathrm{CI}}_{int}=\left(\frac{\ln 2}{t_{\frac{1}{2}}}\right)\left(\frac{350}{0.175}\right)& \left\{\mathrm{Equation}2\right\}\end{array}$

[0385] A number of compounds of formula (I) were tested in this assay, and the results are shown in Table 3 below. 4-Octyl itaconate was included as a comparator compound.

TABLE-US-00003 TABLE 3 Hepatocyte stability Compound Species CI.sub.int (L min.sup.1 10.sup.6 cells) T (min) 4-octyl itaconate Human 401 4 Mouse 351 4 Example 1 Human <3 >460 Mouse <3 >460 Example 2 Human 9 126 Mouse <3 >460 Example 3 Human <3 >460 Mouse <3 >460 Example 4 Human 6 314 Mouse 8 219 Example 5 Human <3 >460 Mouse <3 >460 Example 6 Human 13 143 Mouse <3 >460 Example 7 Human 8 219 Mouse 11 118 Example 8 Human >460 <3 Mouse >460 <3 Example 9 Human 27 44 Mouse 449 3 Example 10 Human 98 12 Mouse >460 <3 Example 11 Human 9 139 Mouse 162 9 Example 12 Human 59 25 Mouse 138 9 Example 13 Human 126 12 Mouse 516 4 Example 14 Human 40 38 Mouse 108 12 Example 15 Human 92 14 Mouse 508 3 Example 16 Human 114 11 Mouse >460 <3 4-octyl itaconate Human 401 4 Mouse 351 4 Example 18 Human 31 52 Mouse 92 14 Example 19 Human 32 48 Mouse 161 13 Example 20 Human 85 18 Mouse 668 3 Example 21 Human 37 42 Mouse 100 21 Example 22 Human 7 204 Mouse 392 4 Example 23 Human <3 >460 Mouse <3 >460

[0386] The results indicate that the compounds of the invention, at least those of Table 3, are expected to have acceptable or improved metabolic stabilities, as shown by their intrinsic clearance (CI.sub.int) and half-life (T.sub.1/2) values, in this assay. All compounds in Table 3, except Example 8, were more stable, i.e., they exhibited lower intrinsic clearance (CI.sub.int) and longer half-life (T.sub.1/2) values compared with 4-octyl itaconate in at least human or mouse species. Preferred compounds exhibited lower intrinsic clearance (CI.sub.int) and longer half-life (T.sub.1/2) values compared with 4-octyl itaconate in both human and mouse species.

REFERENCES

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MISCELLANEOUS

[0440] All references referred to in this application, including patent and patent applications, are incorporated herein by reference to the fullest extent possible.

[0441] Throughout the specification and the claims which follow, unless the context requires otherwise, the word comprise, and variations such as comprises and comprising, will be understood to imply the inclusion of a stated integer, step, group of integers or group of steps but not to the exclusion of any other integer, step, group of integers or group of steps.

[0442] The application of which this description and claims forms part may be used as a basis for priority in respect of any subsequent application. The claims of such subsequent application may be directed to any feature or combination of features described herein. They may take the form of product, composition, process, or use claims and may include, by way of example and without limitation, the following claims.