Kinase inhibitors

Abstract

There are provided compounds of formula I, (I): ##STR00001## wherein R.sup.1 to R.sup.5, X.sup.1, X.sup.2, Ar, L, E, A, A1, G and G.sup.1 have meanings given in the description, which compounds have anti-inflammatory activity (e.g., through inhibition of one or more of members of: the family of p38 mitogen-activated protein kinase enzymes; Syk kinase; and members of the Src family of tyrosine kinases) and have use in therapy, including in pharmaceutical combinations, especially in the treatment of inflammatory diseases, including inflammatory diseases of the lung, eye and intestines.

Claims

1. A compound of formula I, ##STR00044## wherein R.sup.1 and R.sup.2 independently represent C.sub.1-4 alkyl or C.sub.3-6 cycloalkyl, or R.sup.1 and R.sup.2 together combine to form C.sub.2-6 alkylene; R.sup.3 represents C.sub.1-2 alkyl; X.sup.1 and X.sup.2 are both N, or X.sup.1 is C and X.sup.2 is either O or S; Ar is phenyl or a 5- or 6-membered heteroaryl group containing one or more heteroatoms selected from the group consisting of N, O and S, which phenyl and heteroaryl groups are optionally substituted by one or more substituents selected from the group consisting of halo, C.sub.1-4 alkyl, C.sub.1-4 haloalkyl, C.sub.1-4 alkoxy, hydroxy, amino and cyano; L is a direct bond or C.sub.1-2 alkylene; E represents: (a) H, halo, hydroxy, NR.sup.6aR.sup.6b, cyano, C(O)OR.sup.6c, C(O)NR.sup.6dR.sup.6e, SH, S(O).sub.nR.sup.8, S(O).sub.2NR.sup.6fR.sup.6g, (b) C.sub.1-8 alkyl, C.sub.2-8 alkenyl, C.sub.2-8 alkynyl, C.sub.1-8 alkoxy, which latter four groups are optionally substituted by one or more substituents selected from the group consisting of halo and NR.sup.7aR.sup.7b, (c) C.sub.3-8 cycloalkyl, Het.sup.1 or Ar.sup.1, which latter three groups are optionally substituted by one or more substituents selected from the group consisting of halo, hydroxy and C.sub.1-3 alkyl; R.sup.6a to R.sup.6g independently represent H or C.sub.1-4 alkyl, or any one or more of the pairs R.sup.6a and R.sup.6b, R.sup.6d and R.sup.6e, and R.sup.6f and R.sup.6g, when taken together with the N-atom to which each pair is attached, form a saturated 4- to 7-membered heterocyclic group, which heterocyclic group contains one N atom (the atom to which the pairs of substituents are attached) and, optionally, one or more further heteroatoms selected from the group consisting of O, S and N, and which heterocyclic group is optionally substituted by one or more C.sub.1-2 alkyl groups; R.sup.7a and R.sup.7b, independently on each occurrence, represent H or C.sub.1-4 alkyl, or, together with the N-atom to which they are attached, form a 5- to 7-membered heterocyclic group that is fully saturated, partially unsaturated or fully aromatic and which heterocyclic group contains one N atom (the atom to which R.sup.7a and R.sup.7b are attached) and, optionally, one or more further heteroatoms selected from the group consisting of O, S and N, and which heterocyclic group is optionally substituted by one or more substituents selected from the group consisting of halo, hydroxy, oxo, C.sub.1-4 alkyl and C.sub.1-4 alkoxy; R.sup.8 represents C.sub.1-4 haloalkyl, C.sub.1-4 alkyl, C.sub.3-8 cycloalkyl or Ar.sup.2, which latter three groups are optionally substituted by one or more substituents selected from the group consisting of halo, hydroxy and C.sub.1-3 alkyl; Ar.sup.1 and Ar.sup.2 independently represent C.sub.6-14 carbocyclic aryl groups, which groups may be monocyclic, bicyclic or tricyclic and which groups contain at least one ring which is fully aromatic, n is 0, 1 or 2; R.sup.4 and R.sup.5 are each independently C.sub.1-3 alkyl, C.sub.1-3 haloalkyl, cyano or halo, or R.sup.4 and R.sup.5 together combine to form C.sub.3-5 alkylene or C.sub.3-5 alkenylene, which latter two groups are optionally substituted by one or more substituents selected from the group consisting of C.sub.1-3 alkyl, C.sub.1-3 haloalkyl, cyano and halo, or R.sup.4 and R.sup.5, together with the C-atoms to which they are attached, form a fused phenyl or Het.sup.2 ring, which latter two rings are optionally substituted by one or more substituents selected from the group consisting of C.sub.1-3 alkyl, C.sub.1-3 haloalkyl, cyano and halo; Het.sup.1 and Het.sup.2 independently represent 5- to 7-membered heterocyclic groups that are fully saturated, partially unsaturated or fully aromatic, which heterocyclic groups contain one or more heteroatoms selected from the group consisting of N, O and S; one of A and A.sup.1 represents N and the other represents CH, or both of A and A.sup.1 represent CH; G represents phenyl optionally substituted by one or more Y.sup.1 or Het.sup.3 optionally substituted by one or more Y.sup.2; G.sup.1 represents H; or G and G.sup.1 together combine to form C.sub.3-6 alkylene optionally substituted by one or more substituents selected from the group consisting of halo, hydroxy and C.sub.1-3 alkyl, which latter group is optionally substituted by one or more halo atoms or by hydroxy; each Y.sup.1 is independently selected from the group consisting of halo, hydroxy, cyano, SF.sub.5, —OC(O)NH.sub.2, P(O)R.sup.9aR.sup.9b, J.sup.1-N(R.sup.9c)R.sup.9d, J.sup.2-S(O).sub.2R.sup.9e, J.sup.3-[CH.sub.2(CH.sub.2).sub.0-1CH.sub.2—O].sub.2-8—R.sup.9f, —C≡C—R.sup.9g, —N═S(O)R.sup.9hR.sup.9i, Het.sup.a, C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl, C.sub.1-6 alkoxy, C.sub.3-6 cycloalkoxy, —S(O).sub.0-1—C.sub.1-6 alkyl and —S(O).sub.0-1—C.sub.3-6 cycloalkyl which latter six groups are optionally substituted by one or more substituents selected from the group consisting of halo, hydroxy, C.sub.1-3 alkyl, C.sub.1-3 alkoxy and C.sub.3-6 cycloalkyl; each Y.sup.2 independently represents oxo or Y.sup.1; J.sup.1 represents a direct bond, —C(O)— —[C(O)].sub.p—C.sub.1-8 alkylene, —C(O)NR.sup.10a—CH.sub.2—[C.sub.1-7 alkylene]-, -Q.sup.1-CH.sub.2—[C.sub.1-5 alkylene]-, the alkylene parts of which latter four groups are optionally substituted by one or more substituents selected from the group consisting of halo, C.sub.1-3 alkyl and hydroxy; J.sup.2 represents a direct bond, —O—, —NH— C.sub.1-6 alkylene or -Q.sup.2-CH.sub.2—[C.sub.1-5 alkylene]-, the alkylene parts of which latter two groups are optionally substituted by one or more substituents selected from the group consisting of halo, C.sub.1-3 alkyl and hydroxy; J.sup.3 represents —O— or S(O).sub.0-2; Q.sup.1 and Q.sup.2 independently represent O or S(O).sub.0-2; p represents 0 or 1; R.sup.9a and R.sup.9b independently represent C.sub.1-3 alkyl or C.sub.1-3 alkoxy, or R.sup.9a and R.sup.9b together combine to form C.sub.4-6 alkylene; R.sup.9c and R.sup.9d independently represent H or C.sub.1-8 alkyl, which latter group is optionally substituted by R.sup.10b and/or one or more substituents selected from the group consisting of halo and hydroxy; or R.sup.9c and R.sup.9d, together with the N-atom to which they are attached, form a 4- to 7-membered heterocyclic group that is fully saturated, partially unsaturated or fully aromatic and which heterocyclic group contains one N atom (the atom to which R.sup.9c and R.sup.9d are attached) and, optionally, one or more further heteroatoms selected from the group consisting of O, S and N, and which heterocyclic group is optionally substituted by one or more substituents selected from the group consisting of halo, hydroxy, oxo, C.sub.1-4 alkyl and C.sub.1-4 alkoxy; R.sup.9e represents C.sub.1-6 alkyl, C.sub.3-6 cycloalkyl or phenyl, which latter three groups are optionally substituted by one or more substituents selected from the group consisting of halo, hydroxy, C.sub.1-3 alkyl, C.sub.1-3 alkoxy and C.sub.3-6 cycloalkyl; R.sup.9f, R.sup.9g, R.sup.9h and R.sup.9i independently represent C.sub.1-4 alkyl optionally substituted by one or more halo atoms, or R.sup.9f and R.sup.9g independently represent H; R.sup.10a represents H or C.sub.1-3 alkyl optionally substituted by one or more halo atoms; R.sup.10b represents C.sub.1-4 alkoxy, S—C.sub.1-4 alkyl, phenyl or Het.sup.4, which latter two groups are optionally substituted by one or more substituents selected from the group consisting of halo, C.sub.1-4 alkyl, C.sub.1-4 haloalkyl, C.sub.1-4 alkoxy, hydroxy, amino and cyano; Het.sup.3 represents a 5- to 10-membered heteroaromatic group, which group is monocyclic or bicyclic and contains at least one carbocyclic or heterocyclic ring that is fully aromatic, and which group contains one or more heteroatoms selected from the group consisting of N, O and S; Het.sup.4 represents a 4- to 10-membered heterocyclic group that is fully saturated, partially unsaturated or fully aromatic, which heterocyclic group contains one or more heteroatoms selected from the group consisting of N, O and S; and Het.sup.a represents a 5- or 6-membered heterocyclic group that is fully saturated, partially unsaturated or fully aromatic, which group contains one or more heteroatoms selected from the group consisting of N, O and S, and which group is optionally substituted by one or more substituents selected from the group consisting of halo, hydroxy, C.sub.1-3 alkyl, C.sub.1-3 alkoxy and C.sub.3-6 cycloalkyl, or a pharmaceutically acceptable salt, solvate or isotopic derivative thereof.

2. A compound as claimed in claim 1 which is a compound of formula Ia, ##STR00045## wherein R.sup.1 represents C.sub.1-4 alkyl; R.sup.2 and R.sup.3 independently represent C.sub.1-2 alkyl; A.sup.2 represents N or, particularly, CH; R.sup.a represents C.sub.1-2 alkyl or C.sub.1-2 alkoxy, which latter two groups are optionally substituted by one or more halo atoms, or R.sup.a represents C.sub.1-2 alkyl or C.sub.2 alkoxy, which latter two groups are substituted by NR.sup.7aR.sup.7b; R.sup.7a and R.sup.7b both represent C.sub.1-2 alkyl or, together with the N-atom to which they are attached, form a 5- or 6-membered heterocyclic group that is fully saturated and which heterocyclic group contains one N atom (the atom to which R.sup.7a and R.sup.7b are attached) and, optionally, one further heteroatom selected from the group consisting of O, S and N, and which heterocyclic group is optionally substituted by one or more substituents selected from the group consisting of halo, hydroxy, oxo, C.sub.1-4 alkyl and C.sub.1-4 alkoxy, R.sup.4 and R.sup.5 are both halo, or, together with the C-atoms to which they are attached, form a fused phenyl ring; A represents CH or N; A.sup.3 and A.sup.4 both represent CH, or one of A.sup.3 and A.sup.4 represents N and the other represents CH; R.sup.b, R.sup.c and R.sup.d independently represent H, halo, hydroxy, C.sub.1-4 alkylene-N(R.sup.9c)R.sup.9d, —C(O)NH—CH.sub.2—[C.sub.1-5 alkylene]-N(R.sup.9c)R.sup.9d, -Q.sup.1-CH.sub.2—[C.sub.1-3 alkylene]-N(R.sup.9c)R.sup.9d, —O—[CH.sub.2CH.sub.2O].sub.2-7—R.sup.9f, C.sub.1-4 alkyl, C.sub.1-4 alkoxy or C(O)NHC.sub.1-6 alkyl, which latter three groups are optionally substituted by one or more substituents selected from the group consisting of halo and hydroxy, or R.sup.b and R.sup.c, together with the C-atoms to which they are attached, form a fused, 5- or 6-membered aromatic, heteroaromatic or heterocyclic ring, which ring: (i) when heteroaromatic or heterocyclic contains one to three heteroatoms selected from the group consisting of N, O and S; and (ii) is optionally substituted by one or more substituents selected from the group consisting of H, halo, hydroxy, oxo, amino, C.sub.1-2 alkyl and C.sub.1-2 alkoxy, which latter two groups are optionally substituted by one or more halo atoms; R.sup.9c and R.sup.9d both represent C.sub.1-2 alkyl or, together with the N-atom to which they are attached, form a 5- or 6-membered heterocyclic group that is fully saturated and which heterocyclic group contains one N atom (the atom to which R.sup.9c and R.sup.9d are attached) and, optionally, one further heteroatom selected from the group consisting of O, S and N, and which heterocyclic group is optionally substituted by one or more substituents selected from the group consisting of halo, hydroxy, oxo, C.sub.1-4 alkyl and C.sub.1-4 alkoxy, R.sup.9f represents H or methyl, or a pharmaceutically acceptable salt, solvate or isotopic derivative thereof.

3. A compound as claimed in claim 1 which is a compound of formula Ib, ##STR00046## or a pharmaceutically acceptable salt, solvate or isotopic derivative thereof, wherein R.sup.1 to R.sup.5 and A.sup.2 are as defined in claim 1, and R.sup.a represents C.sub.1-2 alkyl or C.sub.1-2 alkoxy, which latter two groups are optionally substituted by one or more halo atoms, or R.sup.a represents C.sub.1-2 alkyl or C.sub.2 alkoxy, which latter two groups are substituted by NR.sup.7aR.sup.7b; R.sup.b, R.sup.c and R.sup.d independently represent H, halo, hydroxy, C.sub.1-4 alkylene-N(R.sup.9c)R.sup.9d, —C(O)NH—CH.sub.2—[C.sub.1-5 alkylene]-N(R.sup.9c)R.sup.9d, -Q.sup.1-CH.sub.2—[C.sub.1-3 alkylene]-N(R.sup.9c)R.sup.9d, —O—[CH.sub.2CH.sub.2O].sub.2-7—R.sup.9f, C.sub.1-4 alkyl, C.sub.1-4 alkoxy or C(O)NHC.sub.1-6 alkyl, which latter three groups are optionally substituted by one or more substituents selected from the group consisting of halo and hydroxy, or R.sup.b and R.sup.c, together with the C-atoms to which they are attached, form a fused, 5- or 6-membered aromatic, heteroaromatic or heterocyclic ring, which ring: (i) when heteroaromatic or heterocyclic contains one to three heteroatoms selected from the group consisting of N, O and S; and (ii) is optionally substituted by one or more substituents selected from the group consisting of H, halo, hydroxy, oxo, amino, C.sub.1-2 alkyl and C.sub.1-2 alkoxy, which latter two groups are optionally substituted by one or more halo atoms; and A.sup.3 and A.sup.4 both represent CH, or one of A.sup.3 and A.sup.4 represents N and the other represents CH.

4. A compound as claimed in claim 1, wherein R.sup.1, R.sup.2 and R.sup.3 are all methyl.

5. A compound as claimed in claim 2, wherein A.sup.2 represents CH.

6. A compound as claimed in claim 2, wherein R.sup.a represents methyl, methoxy or ethoxy, which latter group is optionally substituted by NR.sup.7aR.sup.7b.

7. A compound as claimed in claim 2, wherein: (i) R.sup.b, R.sup.c and R.sup.d are all H, (ii) one of R.sup.b, R.sup.c and R.sup.d is C.sub.1-2 alkylene-N(R.sup.9c)R.sup.9d, —O—CH.sub.2CH.sub.2—N(R.sup.9c)R.sup.9d, —C(O)NH—CH.sub.2CH.sub.2—N(R.sup.9c) R.sup.9d or —O—[CH.sub.2CH.sub.2O].sub.2-7—CH.sub.3, and the other two of R.sup.b, R.sup.c and R.sup.d are selected from the group consisting of H, —C≡C—H, C.sub.1-2 alkyl and C.sub.1-2 alkoxy, which latter two groups are optionally substituted by one or more halo atoms, or (iii) R.sup.d is H or methyl and R.sup.b and R.sup.c, together with the C-atoms to which they are attached, form a fused ring selected from the group consisting of pyrrolidinone, pyrazole and isoxazole, which ring is optionally substituted by amino.

8. A compound as claimed in claim 1, wherein N(R.sup.9c)R.sup.9d represents dimethylamino or morpholin-4-yl.

9. A compound as claimed in claim 1, wherein the structural fragment ##STR00047## represents a group selected from the group consisting of: ##STR00048## wherein R.sup.c and R.sup.d are as defined in claim 2 and A.sup.x represents CH or N.

10. A compound as claimed in claim 1 which is a compound of formula Ic or Id, ##STR00049## or a pharmaceutically acceptable salt, solvate or isotopic derivative thereof, wherein: R.sup.1 to R.sup.5 and A.sup.2 are as defined in claim 1; R.sup.a represents C.sub.1-2 alkyl or C.sub.1-2 alkoxy, which latter two groups are optionally substituted by one or more halo atoms, or R.sup.a represents C.sub.1-2 alkyl or C.sub.2 alkoxy, which latter two groups are substituted by NR.sup.7aR.sup.7b, wherein R.sup.7a and R.sup.7b both represent C.sub.1-2 alkyl or, together with the N-atom to which they are attached, form a 5- or 6-membered heterocyclic group that is fully saturated and which heterocyclic group contains one N atom (the atom to which R.sup.7a and R.sup.7b are attached) and, optionally, one further heteroatom selected from the group consisting of O, S and N, and which heterocyclic group is optionally substituted by one or more substituents selected from the group consisting of halo, hydroxy, oxo, C.sub.1-4 alkyl and C.sub.1-4 alkoxy; and R.sup.c, R.sup.d1 and R.sup.d2 independently represent H, halo, hydroxy, C.sub.1-4 alkylene-N(R.sup.9c)R.sup.9d, —C(O)NH—CH.sub.2—[C.sub.1-5 alkylene]-N(R.sup.9c)R.sup.9d, -Q.sup.1-CH.sub.2—[C.sub.1-3 alkylene]-N(R.sup.9c)R.sup.9d, —O—[CH.sub.2CH.sub.2O].sub.2-7—R.sup.9f, C.sub.1-4 alkyl, C.sub.1-4 alkoxy or C(O)NHC.sub.1-6 alkyl, which latter three groups are optionally substituted by one or more substituents selected from the group consisting of halo and hydroxy, wherein R.sup.9c and R.sup.9d both represent C.sub.1-2 alkyl or, together with the N-atom to which they are attached, form a 5- or 6-membered heterocyclic group that is fully saturated and which heterocyclic group contains one N atom (the atom to which R.sup.9c and R.sup.9d are attached) and, optionally, one further heteroatom selected from the group consisting of O, S and N, and which heterocyclic group is optionally substituted by one or more substituents selected from the group consisting of halo, hydroxy, oxo, C.sub.1-4 alkyl and C.sub.1-4 alkoxy, and R.sup.9f represents H or methyl.

11. A compound as claimed in claim 10, wherein: R.sup.1, R.sup.2 and R.sup.3 are all methyl; A.sup.2 represents CH; R.sup.a represents methyl, methoxy or —O—CH.sub.2CH.sub.2—NR.sup.7aR.sup.7b; NR.sup.7aR.sup.7b represents dimethylamino or, particularly, morpholin-4-yl; R.sup.4 and R.sup.5 either both represent chloro or, together with the C-atoms to which they are attached, form a fused phenyl ring; R.sup.c2 represents H, methoxy, ethoxy, —O—[CH.sub.2CH.sub.2O].sub.2-7—CH.sub.3, —C(O)NH—CH.sub.2CH.sub.2—N(R.sup.9c)R.sup.9d or —O—CH.sub.2CH.sub.2—N(R.sup.9c)R.sup.9d; R.sup.d1 represents H or methyl; R.sup.d2 represents H, methyl, —C≡C—H, trifluoromethyl, methoxy or trifluoromethoxy; and N(R.sup.9c)R.sup.9d represents dimethylamino or, particularly, morpholin-4-yl.

12. A compound as claimed in claim 1, which compound is selected from the group consisting of: 1-(4-((2-(Phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea; 1-(1-(6-Methoxypyridin-3-yl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)-3-(4-((2-(phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)urea; 1-(1-(4-(2-Morpholinoethoxy)phenyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)-3-(4-((2-(phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)urea; 1-(4-((2-(Phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(triethylsilyl)-1H-pyrazol-5-yl)urea; 1-(3-(tert-Butyldimethylsilyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-((2-(phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)urea; 1-(4-((2-((3-(2-Morpholinoethoxy)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea; 1-[4-[2-(1H-Indazol-5-ylamino)pyrimidin-4-yl]oxy-1-naphthyl]-3-[2-(p-tolyl)-5-trimethylsilyl-pyrazol-3-yl]urea; 1-(4-((2-((3-Aminobenzo[d]isoxazol-5-yl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea; 1-(4-((2-((7-Methyl-1H-indazol-5-yl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea; 1-(4-((2-((2-Oxoindolin-6-yl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea; 1-(4-((2-((3-(2-(Dimethylamino)ethoxy)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea; 3-Methoxy-N-(2-morpholinoethyl)-5-((4-((4-(3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)benzamide; 1-(4-((2-((3-Methoxy-5-(2-morpholinoethoxy)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea; 1-(4-((2-(Pyridin-2-ylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea; 1-(4-((2-((7-Methyl-1H-indazol-5-yl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(4-(2-morpholinoethoxy)phenyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea; 1-(3-(Ethyldimethylsilyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-((2-(phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)urea; 1-(2,3-Dichloro-4-((2-((7-methyl-1H-indazol-5-yl)amino)pyrimidin-4-yl)oxy)phenyl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea; 1-(2,3-Dichloro-4-((2-((3-methoxy-5-(2-morpholinoethoxy)phenyl)amino)pyrimidin-4-yl)oxy)phenyl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea; 3-Methoxy-N-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-5-((4-((4-(3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-benzamide; 1-(4-((2-((1-Oxoisoindolin-5-yl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea; 1-(4-((2-((3-Methoxy-5-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea; and 3-Ethynyl-N-(2-morpholinoethyl)-5-((4-((4-(3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)benzamide, or a pharmaceutically acceptable salt, solvate or isotopic derivative thereof.

13. A pharmaceutical formulation comprising a compound as defined in claim 1, or pharmaceutically acceptable salt, solvate or isotopic derivative thereof, in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier.

14. A combination product comprising (A) a compound as defined in claim 1, or pharmaceutically acceptable salt, solvate or isotopic derivative thereof, and (B) another therapeutic agent, wherein each of components (A) and (B) is formulated in admixture with a pharmaceutically-acceptable adjuvant, diluent or carrier.

15. A method of treating inflammation, said method comprising administering to a subject an effective amount of a compound as defined in claim 1, or pharmaceutically acceptable salt, solvate or isotopic derivative thereof, or a pharmaceutical formulation comprising a compound as defined in claim 1, or pharmaceutically acceptable salt, solvate or isotopic derivative thereof, in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier, or a combination product comprising (A) a compound as defined in claim 1, or pharmaceutically acceptable salt, solvate or isotopic derivative thereof, and (B) another therapeutic agent, wherein each of components (A) and (B) is formulated in admixture with a pharmaceutically-acceptable adjuvant, diluent or carrier, wherein the inflammation is a component in a disease selected from the group consisting of cystic fibrosis, pulmonary hypertension, lung sarcoidosis, idiopathic pulmonary fibrosis, Chronic Obstructive Pulmonary Disease (COPD), chronic bronchitis, and emphysema, asthma, paediatric asthma, atopic dermatitis, allergic dermatitis, contact dermatitis or psoriasis, allergic rhinitis, rhinitis, sinusitis, conjunctivitis, allergic conjunctivitis, keratoconjunctivitis sicca (dry eye), glaucoma, diabetic retinopathy, macular oedema, diabetic macular oedema, central retinal vein occlusion (CRVO), dry and/or wet age related macular degeneration (AMD), post-operative cataract inflammation, uveitis, posterior uveitis, anterior uveitis, pan uveitis, corneal graft and limbal cell transplant rejection, gluten sensitive enteropathy (coeliac disease), eosinophilic esophagitis, intestinal graft versus host disease, Crohn's disease and ulcerative colitis.

16. A method according to claim 15, wherein the disease is asthma or COPD.

17. A method according to claim 15, wherein the disease is uveitis, Crohn's disease or ulcerative colitis.

18. A process for the preparation of a compound of formula I according to claim 1, which process comprises: (a) reaction of a compound of formula II, ##STR00050## with a compound of formula III, ##STR00051## wherein one of Z.sup.1 and Z.sup.2 is a structural fragment of formula IV ##STR00052## and the other of Z.sup.1 and Z.sup.2 is a structural fragment of formula V ##STR00053## where E, L, Ar, X.sup.1, X.sup.2, R.sup.1 to R.sup.5, A, A.sup.1, G and G.sup.1 are as defined in claim 1; (b) reaction of a compound of formula IIa, ##STR00054## wherein Z.sup.1 is as defined above, with a suitable azide-forming agent, which reaction is followed, without isolation, by thermal rearrangement of the intermediate acyl azide (of formula Z.sup.1—C(O)—N.sub.3) to provide, in situ, a compound of formula II, which compound is then reacted with a compound of formula III as defined above; (c) reaction of a compound of formula IIb, ##STR00055## wherein LG.sup.1 represents a leaving group and Z.sup.1 is as defined above, with a compound of formula III, as defined above; (d) reaction of a compound of formula VI, ##STR00056## wherein LG.sup.2 represents a leaving group and E, L, Ar, X.sup.1, X.sup.2, R.sup.1 to R.sup.5, A and A.sup.1 are as defined in claim 1, with a compound of formula VII, ##STR00057## wherein G and G.sup.1 are as defined in claim 1; or (e) deprotection of an protected derivative of a compound of formula I, wherein the protected derivative bears a protecting group on an O- or N-atom of the compound of formula I.

Description

PREPARATION OF COMPOUNDS OF THE INVENTION

Example 1

1-(4-((2-(Phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(1) ##STR00022##

(i) 4-((4-Aminonaphthalen-1-yl)oxy)-N-phenylpyrimidin-2-amine

(2) p-TSA monohydrate (2.80 g, 14.72 mmol) was added to a stirred mixture of 4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-amine (see, for example, Cirillo, P. F. et al., WO 2002/92576, 21 Nov. 2000; 8 g, 29.4 mmol) and aniline (6.71 mL, 73.6 mmol) in THF (50 mL) at rt under N.sub.2. The mixture was heated under reflux for 2 h (reaction mixture solidified), a further 50 mL of THF was added and the mixture heated for a further 2 h. The mixture was cooled, diluted with THF (200 mL), the solid filtered and washed with THF (150 mL). The solid was suspended in DCM (100 mL) and 2 M NaOH (35 mL) and the mixture stirred vigorously for 1 h, during which time the solid dissolved. The organic layer was separated, the aq. layer extracted with DCM (100 mL) and the organics combined, dried (MgSO.sub.4) and evaporated under reduced pressure. The residue was triturated with ether and filtered to give the sub-title compound (4.18 g).

(3) 1H NMR (400 MHz; DMSO-d6) δ 9.46 (s, 1H), 8.32 (d, 1H), 8.18-8.15 (m, 1H), 7.64-7.62 (m, 1H), 7.45-7.40 (m, 2H), 7.35 (d, 2H), 7.11 (d, 1H), 7.03-6.99 (m, 2H), 6.80 (t, 1H), 6.72 (d, 1H), 6.42 (d, 1H), 5.79 (s, 2H).

(4) LCMS m/z 329 (M+H).sup.+ (ES.sup.+)

(ii) 1-(4-((2-(Phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(5) DPPA (216 μL, 1.0 mmol) was added to a stirred solution of 1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazole-5-carboxylic acid (see, for example, Barnes, M. J. et al. Bioorg. & Med. Chem. Lett., 17(2), 354-357 (2007) and Bastian, J. A. et al. WO 2007/053394, 10 May 2007; 283 mg, 1.0 mmol) and Et.sub.3N (348 μL, 2.5 mmol) in DMF (5 mL) under N.sub.2 at 0-5° C. After 30 min the mixture was warmed to rt and stirred for a further 1 h. The product from step (i) above (328 mg, 1.0 mmol) was added and the mixture heated at 100° C. for 1 h, cooled and partitioned between EtOAc (100 mL) and water (100 mL). The organic layer was separated, washed with water (100 mL), brine (100 mL), dried (MgSO.sub.4) and evaporated under reduced pressure. The crude product was purified by chromatography on silica gel (40 g column, 0-5% MeOH/DCM) to afford crude product at 70% purity. The solid was triturated with ether (20 mL), filtered and washed with ether (3×20 mL) to afford the title compound (224 mg) as a white solid.

(6) 1H NMR (400 MHz; DMSO-d6) δ 9.51 (s, 1H), 9.16 (s, 1H), 8.76 (s, 1H), 8.40 (d, 1H), 8.09 (d, 1H), 7.94 (d, 1H), 7.82 (d, 1H), 7.64-7.54 (m, 2H), 7.50 (d, 2H), 7.42-7.39 (m, 3H), 7.29 (brd, 2H), 6.99-6.95 (m, 2H), 6.79-6.76 (m, 1H), 6.64 (s, 1H), 6.58 (d, 1H), 2.42 (s, 3H), 0.28 (s, 9H).

(7) LCMS m/z 600 (M+H).sup.+ (ES.sup.+)

Example 2

1-(1-(6-Methoxypyridin-3-yl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)-3-(4-((2-(phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)urea

(8) ##STR00023##

(i) Ethyl 1-(6-methoxypyridin-3-yl)-3-(trimethylsilyl)-1H-pyrazole-5-carboxylate

(9) Pyridine (0.762 mL, 9.42 mmol) followed by activated 4A molecular sieves (3 g) were added to a stirred mixture of (6-methoxypyridin-3-yl)boronic acid (1.441 g, 9.42 mmol), ethyl 3-(trimethylsilyl)-1H-pyrazole-5-carboxylate (1 g, 4.71 mmol) and copper (II) acetate (1.283 g, 7.06 mmol) in DCM (60 mL) at rt. The mixture was stirred for 18 h then filtered through Celite®, and the cake washed with DCM (200 mL). The organics were evaporated under reduced pressure and a mixture of ether (50 mL) and isohexane (50 mL) was added and the solid copper salts filtered. The filtrate was evaporated under reduced pressure and the residue purified by chromatography on silica gel (80 g column, 0-10% EtOAc/isohexane) to afford the sub-title compound (870 mg) as a colourless oil.

(10) 1H NMR (400 MHz; CDCl.sub.3) δ 8.23 (s, 1H), 7.66 (d, 1H), 7.13 (s, 1H), 6.81 (d, 1H), 4.25 (q, 2H), 3.98 (s, 3H), 1.28 (t, 3H), 0.33 (s, 9H).

(11) LCMS m/z 320 (M+H).sup.+ (ES.sup.+)

(ii) 1-(6-Methoxypyridin-3-yl)-3-(trimethylsilyl)-1H-pyrazole-5-carboxylic acid

(12) 2 M NaOH (2019 μL, 4.04 mmol) was added to a solution of the product from step (i) above (860 mg, 2.69 mmol) in EtOH (12 mL) and the mixture stirred at rt for 3 h. The solvent was evaporated under reduced pressure and the residue partitioned between EtOAc (50 mL) and 0.2 M HCl (50 mL). The organic layer was separated, washed with water (2×30 mL), dried (MgSO.sub.4) and evaporated under reduced pressure. The residue was triturated with 5% EtOAc/isohexane to give the sub-title compound (690 mg) as a white solid.

(13) 1H NMR (400 MHz; CDCl.sub.3) δ 8.25 (s, 1H), 7.66 (d, 1H), 7.21 (s, 1H), 6.81 (d, 1H), 3.98 (s, 3H), 0.33 (s, 9H).

(14) LCMS m/z 292 (M+H).sup.+ (ES.sup.+); 290 (M−H).sup.− (ES.sup.−)

(iii) 4-((4-Aminonaphthalen-1-yl)oxy)-N-phenylpyrimidin-2-amine

(15) The sub-title compound can be prepared according to or by analogy with procedures known to those skilled in the art and/or described herein. For example, the following procedure can be used.

(16) p-TSA monohydrate (2.80 g, 14.72 mmol) was added to a stirred mixture of 4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-amine (see, for example, Cirillo, P. F. et al., WO 2002/92576, 21 Nov. 2000; 8 g, 29.4 mmol) and aniline (6.71 mL, 73.6 mmol) in THF (50 mL) at rt under N.sub.2. The mixture was heated under reflux for 2 h (reaction mixture solidified), a further 50 mL of THF was added and the mixture heated for a further 2 h. The mixture was cooled, diluted with THF (200 mL), the solid filtered and washed with THF (150 mL). The solid was suspended in DCM (100 mL) and 2M NaOH (35 mL) and the mixture stirred vigorously for 1 h, during which time the solid dissolved. The organic layer was separated, the aq. layer extracted with DCM (100 mL) and the organics combined, dried (MgSO.sub.4) and evaporated under reduced pressure. The residue was triturated with ether and filtered to give the sub-title compound (4.18 g).

(17) 1H NMR (400 MHz; DMSO-d6) δ 9.46 (s, 1H), 8.32 (d, 1H), 8.18-8.15 (m, 1H), 7.64-7.62 (m, 1H), 7.45-7.40 (m, 2H), 7.35 (d, 2H), 7.11 (d, 1H), 7.03-6.99 (m, 2H), 6.80 (t, 1H), 6.72 (d, 1H), 6.42 (d, 1H), 5.79 (s, 2H).

(18) LCMS m/z 329 (M+H).sup.+ (ES.sup.+)

(iv) 1-(1-(6-Methoxypyridin-3-yl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)-3-(4-((2-(phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)urea

(19) DPPA (111 μL, 0.515 mmol) was added to a stirred solution of the product from step (ii) above (150 mg, 0.515 mmol) and Et.sub.3N (179 μL, 1.287 mmol) in DMF (4 mL) under N.sub.2 at 0-5° C. After 30 min the mixture was warmed to rt and stirred for a further 1 h. 4-((4-Aminonaphthalen-1-yl)oxy)-N-phenylpyrimidin-2-amine (see step (iii) above; 169 mg, 0.515 mmol) was added and the mixture heated at 100° C. for 1 h, cooled and partitioned between EtOAc (50 mL) and water (50 mL). The organic layer was separated, washed with water (50 mL), 20% w/w brine (50 mL), then dried (MgSO.sub.4) filtered and evaporated under reduced pressure to a brown solid. This solid was absorbed onto silica (c.a. 4 g) and purified by chromatography on the Companion (12 g column, 20% EtOAc:isohexane to 100% over 20 CVs) to afford the title compound (155 mg) as a pale yellow solid.

(20) 1H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 9.11 (s, 1H), 8.79 (s, 1H), 8.43 (dd, 1H), 8.39 (d, 1H), 8.05 (d, 1H), 7.94 (dd, 1H), 7.91 (d, 1H), 7.81 (m, 1H), 7.62 (m, 1H), 7.55 (m, 1H), 7.39 (d, 1H), 7.27 (d, 2H), 7.05 (dd, 1H), 6.96 (t, 2H), 6.77 (t, 1H), 6.64 (s, 1H), 6.58 (d, 1H), 3.95 (s, 3H), 0.27 (s, 9H).

(21) LCMS m/z 617 (M+H).sup.+ (ES.sup.+); 615 (M−H).sup.− (ES.sup.−)

Example 3

1-(1-(4-(2-Morpholinoethoxy)phenyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)-3-(4-((2-(phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)urea

(22) ##STR00024##

(i) Ethyl 1-(4-((tert-butyldimethylsilyl)oxy)phenyl)-3-(trimethylsilyl)-1H-pyrazole-5-carboxylate

(23) Pyridine (0.762 mL, 9.42 mmol) followed by activated 4A molecular sieves (1.5 g) were added to a stirred mixture of (4-((tert-butyldimethylsilyl)oxy)phenyl)boronic acid (2.376 g, 9.42 mmol), ethyl 3-(trimethylsilyl)-1H-pyrazole-5-carboxylate (see, for example, Bioorg. & Med. Chem. Left., 17(2), 354-357 (2007); 1 g, 4.71 mmol) and copper (II) acetate (1.283 g, 7.06 mmol) in DCM (50 mL) at rt. The mixture was stirred for 72 h then filtered through Celite®, and the cake washed with DCM (100 mL). The filtrate was evaporated under reduced pressure and the crude product was triturated with ether (80 mL) and filtered. The filtrate was evaporated under reduced pressure and the residue purified by chromatography on silica gel (120 g column, 0-20% ether/isohexane) to afford the sub-title compound (1.9 g) as a colourless oil.

(24) 1H NMR (400 MHz; CDCl.sub.3) δ 7.28 (d, 2H), 7.1 (s, 1H), 6.88 (d, 2H), 4.22 (q, 2H), 1.24 (t, 3H), 1.00 (s, 9H), 0.33 (s, 9H), 0.22 (s, 6H).

(ii) Ethyl 1-(4-hydroxyphenyl)-3-(trimethylsilyl)-1H-pyrazole-5-carboxylate

(25) 1M TBAF in THF (5.78 mL, 5.78 mmol) was added to a stirred solution of the product from step (i) above (2.2 g, 5.25 mmol) in THF (25 mL) at 0-5° C. under N.sub.2. The mixture was stirred for 1.5 h then partitioned between EtOAc (200 mL) and water (100 mL). The organic layer was separated, washed with brine (200 mL), dried (MgSO.sub.4) and evaporated under reduced pressure. The crude product was purified by chromatography on silica gel (120 g column, 0-60% ether/isohexane) to afford the sub-title compound (1.58 g) as a white solid.

(26) 1H NMR (400 MHz; CDCl.sub.3) δ 7.17 (d, 2H), 7.10 (s, 1H), 6.81 (s, 1H), 6.68 (d, 2H), 4.23 (q, 2H), 1.25 (t, 3H), 0.34 (s, 9H).

(27) LCMS m/z 305 (M+H).sup.+ (ES.sup.+); 303 (M−H).sup.− (ES.sup.−)

(iii) Ethyl 1-(4-(2-morpholinoethoxy)phenyl)-3-(trimethylsilyl)-1H-pyrazole-5-carboxylate

(28) K.sub.2CO.sub.3 (681 mg, 4.93 mmol) was added to a mixture of the product from step (ii) above (500 mg, 1.642 mmol) and 4-(2-chloroethyl)morpholine hydrochloride (336 mg, 1.807 mmol) in MeCN (15 mL) at rt. The mixture was heated at 60° C. for 8 h, cooled and partitioned between EtOAc (150 mL) and brine (50 mL). The organic layer was separated, dried (MgSO.sub.4) and evaporated under reduced pressure. The crude product was purified by chromatography on silica gel (40 g column, 0-5% MeOH/DCM) to afford the sub-title compound (685 mg) as a colourless oil.

(29) 1H NMR (400 MHz; CDCl.sub.3) δ 7.34 (d, 2H), 7.1 (s, 1H), 6.96 (d, 2H), 4.23 (q, 2H), 4.15 (t, 2H), 3.76-3.73 (m, 4H), 2.82 (t, 2H), 2.60-2.57 (m, 4H), 1.27 (t, 3H), 0.33 (s, 9H).

(30) LCMS m/z 418 (M+H).sup.+ (ES.sup.+)

(iv) 1-(4-(2-Morpholinoethoxy)phenyl)-3-(trimethylsilyl)-1H-pyrazole-5-carboxylic acid

(31) 1 M NaOH (4.1 mL, 4.10 mmol) was added to a stirred solution of the product from step (iii) above (855 mg, 2.048 mmol) in EtOH (16 mL). The mixture was stirred at rt for 4 h then evaporated under reduced pressure. The residue was dissolved in water (8 mL) and acidified to pH˜4 with aq. 1M HCl, then evaporated under reduced pressure. The crude product was loaded onto a column of SCX in MeOH/water (1:1). The column was washed with MeOH/water (1:1) then MeOH and then the product was eluted with 0.7 M ammonia in MeOH. The resultant mixture was concentrated in vacuo and the residue triturated with ether, filtered and dried at 50° C. to give the sub-title compound (793 mg) as a white solid.

(32) 1H NMR (400 MHz; DMSO-d6) δ 7.33 (d, 2H), 6.95 (d, 2H), 6.67 (s, 1H), 4.12 (t, 2H), 3.61-3.58 (m, 4H), 2.72 (t, 2H), 0.25 (s, 9H). (4H under DMSO)

(33) LCMS m/z 390 (M+H).sup.+ (ES.sup.+); 388 (M−H).sup.− (ES.sup.−)

(v) 1-(1-(4-(2-Morpholinoethoxy)phenyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)-3-(4-((2-(phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)urea

(34) DPPA (138 μL, 0.642 mmol) was added to a stirred solution of the product from step (iv) above (250 mg, 0.642 mmol) and Et.sub.3N (224 μL, 1.605 mmol) in DMF (5 mL) under N.sub.2 at 0-5° C. After 30 min the mixture was warmed to rt and stirred for a further 1 h. 4-((4-Aminonaphthalen-1-yl)oxy)-N-phenylpyrimidin-2-amine (see, for example, Example 2(iii) above; 211 mg, 0.642 mmol) was added, the mixture heated at 100° C. for 1 h, then the solvent removed under reduced pressure. The residue was partitioned between DCM (50 mL) and water (50 mL), the organic phase separated, washed with brine (50 mL), dried (MgSO.sub.4) and evaporated under reduced pressure. The residue was purified by chromatography on silica gel (40 g column, 0-5% MeOH/DCM) to afford 180 mg of a brown foam that was purified by preparative HPLC (Waters, Acidic (0.1% Formic acid). The product was loaded onto a column of SCX in MeOH/DCM. The column was washed with MeOH and then the product was eluted with 0.7 M ammonia in MeOH. The resultant mixture was concentrated in vacuo to afford the title compound (79 mg) as a pale tan solid.

(35) 1H NMR (400 MHz; DMSO-d6) δ 9.52 (s, 1H), 9.15 (s, 1H), 8.71 (s, 1H), 8.38 (d, 1H), 8.06 (d, 1H), 7.92 (d, 1H), 7.80 (d, 1H), 7.63-7.48 (m, 4H), 7.39 (d, 1H), 7.27 (brd, 2H), 7.14 (d, 2H), 6.96 (t, 2H), 6.77 (t, 1H), 6.61 (s, 1H), 6.58 (d, 1H), 4.16 (t, 2H), 3.59-3.57 (m, 4H), 2.73 (t, 2H). (4H under DMSO peak)

(36) LCMS m/z 715 (M+H).sup.+ (ES.sup.+)

Example 4

1-(4-((2-(phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(triethylsilyl)-1H-pyrazol-5-yl)urea

(37) ##STR00025##

(i) Ethyl 3-(triethylsilyl)-1H-pyrazole-5-carboxylate

(38) Ethyl 2-diazoacetate (85 wt %) (1.362 mL, 11.16 mmol) and triethyl(ethynyl)silane (2 mL, 11.16 mmol) were combined and warmed to 70° C. in a sealed tube behind a blast screen for 20 h. After this time the reaction was diluted with isohexanes (10 mL) and the solid collected by filtration, dried under vacuum to afford the sub-title compound (2.42 g) as a white powder.

(39) 1H NMR (400 MHz; CDCl.sub.3) δ 10.67 (br. s, 1H), 6.98 (s, 1H), 4.41 (q, 2H), 1.40 (t, 3H), 1.00 (m, 9H), 0.82 (m, 6H).

(40) LCMS m/z 255 (M+H).sup.+ (ES.sup.+); 253 (M−H).sup.− (ES.sup.−)

(ii) Ethyl 1-(p-tolyl)-3-(triethylsilyl)-1H-pyrazole-5-carboxylate

(41) Pyridine (1.526 mL, 18.87 mmol) followed by activated 4A molecular sieves (3.5 g) were added to a stirred mixture of p-tolylboronic acid (2.57 g, 18.87 mmol), the product from step (i) above (2.4 g, 9.43 mmol) and copper (II) acetate (2.57 g, 14.15 mmol) in DCM (140 mL) at ambient temperature. The mixture was stirred for a total of 24 h and then filtered through Celite® and washed with DCM (200 mL). Ether (100 mL) was added, and the resulting solid removed by filtration through Celite®, washed through with ether (200 mL), organics combined and solvent removed under reduced pressure. The crude product was purified by chromatography on the Companion (120 g column, 0-5% ether in isohexanes) to afford the sub-title compound (3.22 g) as a white powder.

(42) 1H NMR (400 MHz; CDCl.sub.3) δ 7.31 (m, 2H), 7.24 (m, 2H), 7.09 (s, 1H), 4.23 (q, 2H), 2.40 (s, 3H), 1.26 (t, 3H), 1.01 (m, 9H), 0.83 (m, 6H).

(43) LCMS m/z 345 (M+H).sup.+ (ES.sup.+)

(iii) 1-(p-Tolyl)-3-(triethylsilyl)-1H-pyrazole-5-carboxylic acid

(44) To a solution of the product from step (ii) above (3.22 g, 9.35 mmol) in ethanol (100 mL) was added 2M NaOH (aq) (9.35 mL, 18.69 mmol) and the resulting solution stirred over 16 h at ambient temperature. Solvent was removed under reduced pressure and the residue partitioned between water (100 mL) and ether (150 mL). The aqueous layer was acidified to pH 1 with 1 M HCl and extracted with ether (50 mL). The ether layers were combined and washed with brine (50 mL), dried (MgSO.sub.4) and evaporated under reduced pressure to give the sub-title compound (2.645 g) as a cream coloured powder.

(45) 1H NMR (400 MHz; CDCl.sub.3) δ 8.66 (br.s, 1H), 7.32-7.28 (m, 2H), 7.24-7.19 (br.d, 2H), 7.19 (s, 1H), 2.40 (s, 3H), 1.03-0.97 (m, 9H), 0.86-0.78 (m, 6H).

(46) LCMS m/z 317 (M+H).sup.+ (ES.sup.+); 315 (M−H).sup.− (ES.sup.−)

(iv) 1-(4-((2-(phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(triethylsilyl)-1H-pyrazol-5-yl)urea

(47) To a nitrogen-purged solution of the product from step (iii) above (200 mg, 0.632 mmol) in anhydrous dioxane (3 mL) was added DPPA (204 μL, 0.948 mmol) and then Et.sub.3N (264 μL, 1.896 mmol) and the reaction stirred at ambient temperature for 30 min. 4-((4-Aminonaphthalen-1-yl)oxy)-N-phenylpyrimidin-2-amine (see, for example, Example 2(iii) above; 311 mg, 0.948 mmol) was then added and the reaction was stirred at 70° C. for 30 min and then at 100° C. for a further 90 min. After this time the reaction was cooled to ambient temperature and solvent removed under reduced pressure, and coevaporated with DCM (15 mL). The resulting brown oil was purified by chromatography on silica gel (40 g column, 0-20% EtOAc in toluene) to afford a dark pink solid. Trituration from ether (5 mL) afforded a pale pink solid which was dried under vacuum at 40° C. to give the title compound (84 mg).

(48) 1H NMR (400 MHz; DMSO-d6) δ 9.52 (s, 1H), 9.18 (s, 1H), 8.79 (s, 1H), 8.39 (d, 1H), 8.09 (d, 1H), 7.94 (d, 1H), 7.81 (dd, 1H), 7.62 (ddd, 1H), 7.55 (ddd, 1H), 7.50 (m, 2H), 7.40 (m, 3H), 7.27 (br.d, 2H), 6.96 (t, 2H), 6.76 (t, 1H), 6.62 (s, 1H), 6.58 (d, 1H), 2.41 (s, 3H), 1.00 (t, 9H), 0.76 (q, 6H).

(49) LCMS m/z 642 (M+H).sup.+ (ES.sup.+); 640 (M−H).sup.− (ES.sup.−)

Example 5

1-(3-(tert-Butyldimethylsilyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-((2-(phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)urea

(50) ##STR00026##

(i) Ethyl 3-(tert-butyldimethylsilyl)-1H-pyrazole-5-carboxylate

(51) Ethyl 2-diazoacetate (85 wt %) (1.740 mL, 14.26 mmol) and tert-butyl(ethynyl)-dimethylsilane (2.66 mL, 14.26 mmol) were combined and warmed to 70° C. in a sealed tube behind a blast screen for 20 h. After this time the reaction was diluted with isohexanes (10 mL) and the solid collected by filtration, dried under vacuum to afford the sub-title compound (2.1 g) as a white powder.

(52) 1H NMR (400 MHz; CDCl.sub.3) δ 10.69 (br.s, 1H), 6.99 (s, 1H), 4.41 (q, 2H), 1.41 (t, 3H), 0.92 (s, 9H), 0.32 (s, 6H).

(53) LCMS m/z 255 (M+H).sup.+ (ES.sup.+); 253 (M−H).sup.− (ES.sup.−)

(ii) Ethyl 3-(tert-butyldimethylsilyl)-1-(p-tolyl)-1H-pyrazole-5-carboxylate

(54) Pyridine (1.335 mL, 16.51 mmol) followed by activated 4A molecular sieves (2.5 g) were added to a stirred mixture of p-tolylboronic acid (2.245 g, 16.51 mmol), the product from step (i) above (2.1 g, 8.25 mmol) and copper (II) acetate (2.249 g, 12.38 mmol) in DCM (120 mL) at ambient temperature. The mixture was stirred for 4 days at rt then filtered through Celite®, and the cake washed with DCM (300 mL). The organics were evaporated under reduced pressure and the crude product was purified by chromatography on silica gel (90 g column, 0-10% ether/isohexane) to afford the sub-title compound (1.02 g) as a clear, colourless oil.

(55) 1H NMR (400 MHz; CDCl.sub.3) δ 7.33-7.28 (m, 2H), 7.26-7.22 (m, 2H), 7.10 (s, 1H), 4.23 (q, 2H), 2.40 (s, 3H), 1.27 (t, 3H), 0.96 (s, 9H), 0.30 (s, 6H).

(56) LCMS m/z 345 (M+H).sup.+ (ES.sup.+)

(iii) 3-(tert-Butyldimethylsilyl)-1-(p-tolyl)-1H-pyrazole-5-carboxylic acid

(57) To a solution of the product from step (ii) above (1.02 g, 2.96 mmol) in ethanol (50 mL) was added a solution of 2 M NaOH (aq) (2.96 mL, 5.92 mmol) and the mixture stirred at ambient temperature over 16 h. Solvent was removed under reduced pressure and the residue partitioned between water (50 mL) and ether (100 mL). The aqueous layer was acidified to pH 1 with 1 M HCl and extracted with ether (40 mL). The ether layers were combined and washed with brine (50 mL), dried (MgSO.sub.4) and evaporated under reduced pressure to give the sub-title compound (783 mg) as a cream coloured powder.

(58) 1H NMR (400 MHz; CDCl.sub.3) δ 7.33-7.29 (m, 2H), 7.25-7.21 (br.d, 2H), 7.20 (s, 1H), 2.41 (s, 3H), 0.94 (s, 9H), 0.29 (m, 6H).

(59) LCMS m/z 317 (M+H).sup.+ (ES.sup.+); 315 (M−H).sup.− (ES.sup.−)

(iv) 1-(3-(tert-Butyldimethylsilyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-((2-(phenylamino)-pyrimidin-4-yl)oxy)naphthalen-1-yl)urea

(60) To a nitrogen-purged solution of the product from step (iii) above (200 mg, 0.632 mmol) in anhydrous dioxane (3 mL) was added DPPA (204 μL, 0.948 mmol) and then Et.sub.3N (264 μL, 1.896 mmol) and the reaction stirred at ambient temperature for 30 min. 4-((4-Aminonaphthalen-1-yl)oxy)-N-phenylpyrimidin-2-amine (see, for example, Example 2(iii) above; 208 mg, 0.632 mmol) was added and the reaction was stirred at 70° C. for 30 min and then at 100° C. for a further 90 min. The solvent was removed under reduced pressure and the crude product was purified by chromatography on the Companion (40 g column, 0-25% EtOAc in toluene) to afford crude product as a tan solid. This product was further purified by chromatography on the Companion (12 g column, 0-1.1% (MeOH/1M NH3) in DCM) to afford the title compound (130 mg) as a beige solid.

(61) 1H NMR (400 MHz; DMSO-d6) δ 9.49 (s, 1H), 9.16 (s, 1H), 8.77 (s, 1H), 8.39 (d, 1H), 8.09 (d, 1H), 7.94 (d, 1H), 7.81 (dd, 1H), 7.62 (ddd, 1H), 7.55 (ddd, 1H), 7.51-7.47 (m, 2H), 7.43-7.37 (m, 3H), 7.27 br.d, 2H), 6.96 (t, 2H), 6.76 (t, 1H), 6.63 (s, 1H), 6.57 (d, 1H), 2.41 (s, 3H), 0.95 (s, 9H) 0.23 (s, 6H).

(62) LCMS m/z 642 (M+H).sup.+ (ES.sup.+); 640 (M−H).sup.− (ES.sup.−)

Example 6

1-(4-((2-((3-(2-Morpholinoethoxy)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(63) ##STR00027##

(i) 1-(4-((2-Chloropyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(64) DPPA (377 μL, 1.749 mmol) was added to a stirred solution of 1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazole-5-carboxylic acid (see, for example, Barnes, M. J. et al. Bioorg. & Med. Chem. Lett., 17(2), 354-357 (2007) and Bastian, J. A. et al. WO 2007/053394, 10 May 2007; 480 mg, 1.749 mmol) and Et.sub.3N (610 μL, 4.37 mmol) in DMF (10 mL) at 0-5° C. After 30 min the mixture was warmed to rt and stirred for a further 1 h. 4-((2-Chloropyrimidin-4-yl)oxy)naphthalen-1-amine (see, for example, Cirillo, P. F. et al., WO 2002/92576, 21 Nov. 2000; 452 mg, 1.662 mmol) was added and the mixture heated at 100° C. for 1 h, cooled and partitioned between EtOAc (100 mL) and water (50 mL). The organic layer was separated, washed with water (50 mL), and 20% w/w brine (50 mL), dried (MgSO.sub.4), filtered and evaporated under reduced pressure to a brown solid. The crude product was purified by chromatography on the Companion (40 g column, 3% MeOH:DCM) to afford the sub-title compound (420 mg) as a pale green glass.

(65) 1H NMR (400 MHz, DMSO-d6) δ 9.18 (s, 1H), 8.82 (s, 1H), 8.66 (d, 1H), 8.09 (d, 1H), 7.97 (d, 1H), 7.79 (m, 1H), 7.60 (s, 2H), 7.45 (m, 4H), 7.27 (d, 1H), 6.63 (s, 1H), 5.76 (s, 1H), 2.42 (s, 3H), 0.27 (s, 9H).

(66) LCMS m/z 543/545(M+H).sup.+ (ES.sup.+)

(ii) 1-(4-((2-((3-(2-Morpholinoethoxy)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(67) To a stirred solution of the product from step (i) above (90 mg, 0.166 mmol) in DMF (2 mL) was added p-TSA monohydrate (63.0 mg, 0.331 mmol) then 3-(2-morpholinoethoxy)aniline (73.7 mg, 0.331 mmol) and heated at 65° C. for 16 h. More p-TSA monohydrate (31.0 mg, 0.165 mmol) was added and heating continued for a further 16 h. The mixture was cooled and partitioned between EtOAc (10 mL) and sat. NaHCO.sub.3 solution, the organic layer separated and washed with 20% w/w brine (10 mL), dried (MgSO.sub.4) filtered and solvents evaporated. The crude product was purified by preparative HPLC (Gilson, Acidic (0.1% Formic acid), Waters X-Select Prep-C18, 5 μm, 19×50 mm column, 40%-75% MeCN in Water) to afford the title compound (65 mg) as a colourless solid.

(68) 1H NMR (400 MHz, DMSO-d6) δ 9.47 (s, 1H), 9.20 (s, 1H), 8.84 (s, 1H), 8.39 (d, 1H), 8.08 (d, 1H), 7.96 (d, 1H), 7.81 (m, 1H), 7.59 (m, 2H), 7.49 (m, 2H), 7.40 (m, 3H), 7.13 (s, 1H), 6.94 (m, 1H), 6.88 (m, 1H), 6.63 (s, 1H), 6.55 (d, 1H), 6.41 (m, 1H), 3.90 (t, 2H), 3.53 (t, 4H), 2.60 (t, 2H), 2.41 (m, 7H), 0.27 (s, 9H).

(69) LCMS m/z 729 (M+H).sup.+ (ES.sup.+)

Example 7

1-[4-[2-(1H-Indazol-5-ylamino)pyrimidin-4-yl]oxy-1-naphthyl]-3-[2-(p-tolyl)-5-trimethylsilyl-pyrazol-3-yl]urea

(70) ##STR00028##

(71) To a stirred solution of 1-(4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea (see Example 6(i) above; 100 mg, 0.184 mmol) in DMF (2 mL) was added p-TSA monohydrate (35.0 mg, 0.184 mmol) then 1H-indazol-5-amine (49.0 mg, 0.368 mmol). The mixture was heated at 100° C. for 20 min in a microwave (200 W), cooled and partitioned between EtOAc (10 mL) and sat. NaHCO.sub.3 solution (20 mL). The organic layer separated and washed with 20% w/w brine (20 mL), dried (MgSO.sub.4), filtered and solvents evaporated to give a dark red gum. The crude product was purified by preparative HPLC (Gilson, Acidic (0.1% Formic acid), Waters X-Select Prep-C18, 5 μm, 19×50 mm column, 40%-75% MeCN in Water) to afford the title compound (56 mg) as a colourless solid.

(72) 1H NMR (400 MHz, DMSO-d6) δ 12.77 (s, 1H), 9.53 (s, 1H), 9.28 (s, 1H), 8.91 (s, 1H), 8.39 (d, 1H), 8.13 (d, 1H), 8.06 (d, 1H), 7.83 (m, 1H), 7.56 (m, 6H), 7.42 (m, 3H), 7.24 (s, 2H), 6.64 (s, 1H), 6.58 (d, 1H), 2.41 (s, 3H), 0.27 (s, 9H).

(73) LCMS m/z 640 (M+H).sup.+ (ES.sup.+)

Example 8

1-(4-((2-((3-Aminobenzo[d]isoxazol-5-yl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(74) ##STR00029##

(75) To a stirred solution of 1-(4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea (see Example 6(i) above; 100 mg, 0.184 mmol) in DMF (2 mL) was added p-TSA monohydrate (35 mg, 0.184 mmol) then tert-butyl (5-aminobenzo[d]isoxazol-3-yl)carbamate (92 mg, 0.368 mmol). The mixture was heated at 60° C. for 4 h in a microwave (200 W), cooled and partitioned between EtOAc (20 mL) and sat. NaHCO.sub.3 solution (20 mL). The organic layer was separated and washed with 20% w/w brine (20 mL), dried (MgSO.sub.4), filtered and solvents evaporated to give a brown glass. The crude product was purified by chromatography on the Companion (12 g column, DCM then 2% MeOH:DCM) to afford the BOC protected compound (100 mg) as a pale tan solid which was dissolved in IPA (2 mL) then HCl 5-6M in IPA (402 μL) added and stirred at 60° C. for 4 h. The crude product was purified by preparative HPLC (Gilson, Basic (0.1% Ammonia), Waters X-Select Prep-C18, 5 μm, 19×50 mm column, 35-80% MeCN in Water) to afford the title compound (36 mg) as a colourless solid.

(76) 1H NMR (400 MHz, DMSO-d6) δ 9.62 (s, 1H), 9.19 (s, 1H), 8.83 (s, 1H), 8.36 (d, 1H), 8.07 (d, 1H), 7.91 (d, 2H), 7.81 (m, 1H), 7.60 (m, 2H), 7.49 (m, 2H), 7.41 (dd, 4H), 7.12 (d, 1H), 6.63 (s, 1H), 6.51 (d, 1H), 6.24 (s, 2H), 2.41 (s, 3H), 0.27 (s, 9H).

(77) LCMS m/z 656 (M+H).sup.+ (ES.sup.+)

Example 9

1-(4-((2-((7-Methyl-1H-indazol-5-yl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(78) ##STR00030##

(79) To a stirred solution of 1-(4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea (see Example 6(i) above; 303 mg, 0.558 mmol) in THF/DMF (6 mL, 1:2) was added p-TSA monohydrate (212 mg, 1.116 mmol) then 7-methyl-1H-indazol-5-amine (164 mg, 1.116 mmol). The resulting solution was heated at 60° C. overnight. The reaction was cooled to rt and partitioned between EtOAc (80 mL) and sodium bicarbonate solution (60 mL). The aqueous phase was extracted with EtOAc (2×80 mL). The combined organic extracts were washed with water (2×60 mL), brine (60 mL), dried (MgSO.sub.4), filtered and concentrated to afford a brown solid (448 mg). The crude product was purified by chromatography on silica gel (40 g column, 0-10% MeOH in DCM) to afford a residue which was triturated with MeOH to afford the title compound (158 mg) as a white solid.

(80) 1H NMR (DMSO-d6) 400 MHz, δ: 12.85 (s, 1H) 9.45 (s, 1H) 9.22 (s, 1H) 8.86 (s, 1H) 8.38 (d, 1H) 8.13-8.06 (m, 2H) 7.83-7.81 (m, 1H) 7.64-7.50 (m, 5H) 7.43-7.30 (m, 4H) 7.02 (s, 1H) 6.65 (s, 1H) 6.58 (d, 1H) 2.41 (s, 3H) 2.29 (s, 3H) 0.27 (s, 9H).

(81) LCMS m/z 654 (M+H).sup.+ (ES.sup.+)

Example 10

1-(4-((2-((2-Oxoindolin-6-yl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(82) ##STR00031##

(83) To a stirred solution of 1-(4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea (see Example 6(i) above; 300 mg, 0.552 mmol) in THF/DMF (6 mL, 1:2) was added p-TSA monohydrate (105 mg, 0.552 mmol) then 6-aminoindolin-2-one (164 mg, 1.105 mmol). The resulting solution was heated at 60° C. overnight. The reaction was cooled to rt and partitioned between EtOAc (80 mL) and sodium bicarbonate solution (60 mL). A yellow solid precipitated and this was filtered and purified by chromatography on the Companion (12 g column, 0-5% MeOH in DCM) to afford the title compound (51 mg) as a pale yellow powder.

(84) 1H NMR (400 MHz; DMSO-d6) δ 10.21 (s, 1H), 9.51 (s, 1H), 9.16 (s, 1H), 8.77 (s, 1H), 8.37 (d, 1H), 8.06 (d, 1H), 7.92 (d, 1H), 7.81 (br,dd, 1H), 7.66-7.59 (m, 1H), 7.59-7.53 (m, 1H), 7.51-7.46 (m, 2H), 7.43-7.38 (m, 3H), 7.15 (br.s, 1H), 6.94 (br.d, 1H), 6.79 (br.d, 1H), 6.64 (s, 1H), 6.49 (d, 1H), 3.30 (s, 2H), 2.41 (s, 3H), 0.26 (s, 9H).

(85) LCMS m/z 655 (M+H).sup.+ (ES.sup.+)

Example 11

1-(4-((2-((3-(2-(Dimethylamino)ethoxy)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(86) ##STR00032##

(87) To a stirred solution of 1-(4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea (see Example 6(i) above; 150 mg, 0.276 mmol) in THF/DMF (3 mL, 1:2) was added p-TSA monohydrate (105 mg, 0.552 mmol) followed by 3-(2-(dimethylamino)ethoxy)aniline (100 mg, 0.552 mmol). The resulting mixture was heated at 60° C. overnight. The reaction was cooled to rt and partitioned between EtOAc (30 mL) and sodium bicarbonate solution (20 mL). The aqueous phase was extracted with EtOAc (2×30 mL). The combined organic extracts were washed with water (2×30 mL), brine (30 mL), dried (MgSO.sub.4), filtered and concentrated in vacuo to afford a residue (250 mg). The crude product was purified by chromatography on silica gel (40 g column, 0-10% MeOH in DCM) to afford an off-white solid (100 mg) at 95% purity. The solid was triturated with Et.sub.2O to afford the title compound (73 mg) as a tan solid.

(88) 1H NMR (DMSO-d6) 400 MHz, δ: 9.47 (s, 1H) 9.15 (s, 1H) 8.78 (s, 1H) 8.40 (d, 1H) 8.07 (d, 1H) 7.94 (d, 1H) 7.81 (d, 1H) 7.64-7.60 (m, 1H) 7.57-7.54 (m, 1H) 7.49-7.47 (m, 2H) 7.41-7.39 (m, 3H) 7.12 (br s, 1H) 6.96-6.94 (br m, 1H) 6.88-6.84 (br m, 1H) 6.62 (s, 1H) 6.55 (d, 1H) 6.42-6.40 (m, 1H) 3.86 (t, 2H) 2.54 (t, 2H) 2.41 (s, 3H) 2.17 (s, 6H) 0.26 (s, 9H).

(89) LCMS m/z 687 (M+H).sup.+ (ES.sup.+); 685 (M−H).sup.− (ES.sup.−)

Example 12

3-Methoxy-N-(2-morpholinoethyl)-5-((4-((4-(3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)benzamide

(90) ##STR00033##

(i) 3-Amino-5-methoxy-N-(2-morpholinoethyl)benzamide

(91) To a stirred mixture of 3-amino-5-methoxybenzoic acid (5.20 g, 31.1 mmol), Et.sub.3N (4.50 mL, 32.3 mmol) and 2-morpholinoethanamine (4.23 mL, 32.3 mmol) in THF (150 mL) and DMF (4 mL) was added HATU (14.72 g, 38.7 mmol) and the reaction stirred at ambient temperature overnight. After this time the mixture was taken up in ethyl acetate (300 mL) and washed with saturated NaHCO.sub.3 (aq) (2×100 mL). The aqueous was back extracted with further ethyl acetate (4×50 mL) and organics combined, dried over MgSO.sub.4, filtered and concentrated under reduced pressure. Trituration with isohexanes (100 mL) afforded a pale orange gum (15 g). The crude product was purified by chromatography on the Companion (220 g column, 0-60% IPA in DCM). Fractions were combined as two separate batches to afford the sub-title compound (5.35 g) as an orange solid.

(92) 1H NMR (400 MHz; CDCl.sub.3) δ: 6.69-6.64 (m, 3H), 6.35 (t, 1H), 3.81 (br.s, 2H), 3.81 (s, 3H), 3.73 (m, 4H), 3.53 (dd, 2H), 2.62-2.57 (m, 2H), 2.53-2.49 (m, 4H).

(93) LCMS m/z 280 (M+H).sup.+ (ES.sup.+)

(ii) 3-Methoxy-N-(2-morpholinoethyl)-5-((4-((4-(3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)benzamide

(94) To a stirred solution of 1-(4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea (see Example 6(i) above; 152 mg, 0.280 mmol) in THF/DMF (3 mL, 1:1) was added p-TSA monohydrate (106 mg, 0.560 mmol) followed by the product from step (i) above (94 mg, 0.336 mmol). The resulting mixture was heated at 60° C. overnight. The reaction was cooled to rt and partitioned between EtOAc (30 mL) and sodium bicarbonate solution (20 mL). The aqueous layer was extracted with EtOAc (2×30 mL). The combined organic extracts were washed with water (2×30 mL), brine (30 mL), dried (MgSO.sub.4), filtered and concentrated in vacuo to afford a light pink solid. The crude product was purified by chromatography on silica gel (40 g column, 0-10% MeOH in DCM) to afford an off-white solid which was purified by preparative HPLC (Varian, Acidic (0.1% Formic acid), Agilent Prep C-18, 5 μm, 21.2×50 mm column, 25-70% MeCN in Water) to afford the title compound (62 mg) as a white solid.

(95) 1H NMR (d4-MeOH) 400 MHz, δ: 8.34 (d, 1H) 7.95-7.86 (m, 2H) 7.75 (d, 1H) 7.56-7.49 (m, 2H) 7.38-7.30 (m, 6H) 7.24-7.23 (m, 1H) 6.86-6.85 (m, 1H) 6.68 (s, 1H) 6.56 (d, 1H) 3.71-3.69 (m, 4H) 3.57 (s, 3H) 3.50 (t, 2H) 2.65-2.58 (m, 6H) 2.42 (s, 3H) 0.31 (s, 9H).

(96) LCMS m/z 786 (M+H).sup.+ (ES.sup.+); 784 (M−H).sup.− (ES.sup.−)

Example 13

1-(4-((2-((3-Methoxy-5-(2-morpholinoethoxy)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(97) ##STR00034##

(i) 3-Methoxy-5-(2-morpholinoethoxy)aniline

(98) To a stirred suspension of 3-amino-5-methoxyphenol (205 mg, 1.473 mmol) and K.sub.2CO.sub.3 (1018 mg, 7.37 mmol) in pyridine/DMF (2 mL, 1:3) was added 4-(2-chloroethyl)morpholine hydrochloride (274 mg, 1.473 mmol). The resulting mixture was heated at 60° C. overnight. The reaction was cooled to rt, filtered and concentrated in vacuo to afford a brown oil (850 mg). The crude product was purified by chromatography on silica gel (40 g column, 0-10% MeOH in DCM) to afford the sub-title compound (201 mg) as a sticky orange oil.

(99) 1H NMR (DMSO-d6) 400 MHz, δ: 5.75-5.73 (m, 2H) 5.67 (t, 1H) 5.05 (s, 2H) 3.94 (t, 2H) 3.61 (s, 3H) 3.58-3.55 (m, 4H) 2.62 (t, 2H) 2.45-2.43 (m, 4H).

(100) LCMS m/z 253 (M+H).sup.+ (ES.sup.+)

(ii) 1-(4-((2-((3-Methoxy-5-(2-morpholinoethoxy)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(101) p-TSA monohydrate (106 mg, 0.556 mmol) was added to a stirred solution of 1-(4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea (see Example 6(i) above; 151 mg, 0.278 mmol) and the product from step (i) above (156 mg, 0.556 mmol) in THF/DMF (3 mL, 2:1). The resulting mixture was heated at 60° C. for 5 h. Stirring continued at 60° C. for 24 h. The reaction mixture was cooled to rt then partitioned between EtOAc (30 mL) and sodium bicarbonate solution (20 mL). The aqueous phase was extracted with EtOAc (2×30 mL). The combined organic extracts were washed with water (2×30 mL), brine (30 mL), dried (MgSO.sub.4), filtered and concentrated in vacuo to afford an oil at 65% purity. The crude product was purified by chromatography on silica gel (40 g column) to afford a solid. The crude product was purified by preparative HPLC (Gilson, Acidic (0.1% Formic acid), Waters X-Select Prep-C18, 5 μm, 19×50 mm column, 10-50% MeCN in Water) to afford an oil (100 mg) which was triturated with ether/isohexane to afford the title compound (50 mg) as an off-white solid.

(102) 1H NMR (DMSO-d6) 400 MHz, δ: 9.40 (s, 1H) 9.11 (s, 1H) 8.78 (s, 1H) 8.40 (d, 1H) 8.06 (d, 1H) 7.96 (d, 1H) 7.83-7.80 (m, 1H) 7.64-7.60 (m, 1H) 7.58-7.54 (m, 1H) 7.49-7.47 (m, 2H) 7.41-7.38 (m, 3H) 6.79-6.77 (m, 2H) 6.62 (s, 1H) 6.53 (d, 1H) 6.03-6.02 (br m, 1H) 3.88 (br t, 2H) 3.55-3.53 (br m, 4H) 3.49 (s, 3H) 2.59 (br t, 2H) 2.45-2.35 (br m, 7H) 0.26 (s, 9H).

(103) LCMS m/z 759 (M+H).sup.+ (ES.sup.+); 757 (M−H).sup.− (ES.sup.−)

Example 14

1-(4-((2-(pyridin-2-ylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(104) ##STR00035##

(i) 4-((4-Aminonaphthalen-1-yl)oxy)-N-(pyridin-2-yl)pyrimidin-2-amine

(105) A sealed tube was charged with BINAP (33.5 mg, 0.054 mmol), Pd.sub.2dba.sub.3 (39.4 mg, 0.043 mmol), caesium carbonate (105 mg, 0.323 mmol), tert-butyl (4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)carbamate (see, for example, Ito, K. et al., WO 2010/067130, 17 Jun. 2010; 80 mg, 0.215 mmol) and pyridin-2-amine (25.3 mg, 0.269 mmol) and anhydrous DMA (1 mL) was added and the mixture purged with nitrogen for 10 mins. The reaction was then heated overnight at 80° C. Reaction performed in duplicate. The crude product was loaded onto a column of SCX (0.5 g) in MeOH-DCM 1:1 (1 mL). The column was washed with MeOH (5 mL) and then the product was eluted with 0.7 M ammonia in MeOH. The resultant mixture was concentrated in vacuo to afford crude product as a brown oil. This was dissolved in TFA (3 mL) and DCM (3 mL) and stirred over 16 h. After this time the crude mixture was loaded on to SCX (1 g). The column was washed with MeOH (10 mL) and then the product was eluted with 0.7 M ammonia in MeOH. The resultant mixture was concentrated in vacuo to afford crude product as a brown oil. The crude product was purified by chromatography on the Companion (12 g column, 0-5% (1 N NH.sub.3 in MeOH) in DCM to afford the sub-title compound (57 mg) as a dark brown oil.

(ii) 1-(4-((2-(pyridin-2-ylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(106) To a nitrogen-purged solution of 1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazole-5-carboxylic acid (see, for example, Barnes, M. J. et al. Bioorg. & Med. Chem. Lett., 17(2), 354-357 (2007) and Bastian, J. A. et al. WO 2007/053394, 10 May 2007; 49.2 mg, 0.179 mmol) in anhydrous dioxane (1 mL) was added DPPA (57.9 μL, 0.269 mmol) and then Et.sub.3N (74.9 μL, 0.537 mmol) and the reaction stirred at ambient temperature for 30 min. The product from step (i) above (59 mg, 0.179 mmol) was added and the reaction was stirred at 100° C. for a further 90 mins. Solvent was removed under reduced pressure and the crude material was purified by chromatography on the Companion (12 g column, 0-5% (1 M NH.sub.3 in MeOH) in DCM) to afford a brown solid (16 mg). Trituration with ether (3 mL) afforded the title compound as a pale brown solid (10 mg).

(107) 1H NMR (400 MHz; DMSO-d6) δ 9.75 (s, 1H), 9.18 (s, 1H), 8.77 (s, 1H), 8.44 (d, 1H), 8.12 (dq, 1H), 8.08 (d, 1H), 7.94 (d, 1H), 7.80 (dd, 1H), 7.62 (ddd, 1H), 7.55 (ddd, 1H), 7.49 (m, 2H), 7.42 (m, 3H), 7.25 (br.d, 1H), 7.17 (m, 1H), 6.80 (ddd, 1H), 6.67 (d, 1H), 6.63 (s, 1H), 2.41 (s, 3H), 0.27 (s, 9H).

(108) LCMS m/z 601 (M+H).sup.+ (ES.sup.+)

Example 15

1-(4-((2-((7-Methyl-1H-indazol-5-yl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(4-(2-morpholinoethoxy)phenyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(109) ##STR00036##

(i) 1-(4-((2-Chloropyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(4-(2-morpholinoethoxy)phenyl)-3-(trimethylsilyl)-1H-yl)pyrazol-5-yl)urea

(110) To a stirred solution of 1-(4-(2-morpholinoethoxy)phenyl)-3-(trimethylsilyl)-1H-pyrazole-5-carboxylic acid (see Example 3(iv) above; 527 mg, 1.353 mmol) and Et.sub.3N (471 μL, 3.38 mmol) in DMF (8 mL) at 0-5° C. was added DPPA (306 μL, 1.421 mmol). After 30 min the ice bath was removed and the reaction allowed to warm to rt. Stirring continued at rt for 1 h. 4-((2-Chloropyrimidin-4-yl)oxy)naphthalen-1-amine (see, for example, Cirillo, P. F. et al., WO 2002/92576, 21 Nov. 2000; 368 mg, 1.353 mmol) was added and the reaction heated at 100° C. for 2 h. The reaction was cooled to rt and partitioned between EtOAc (100 mL) and water (100 mL). The organic phase was washed with water (100 mL), brine (100 mL), dried (MgSO.sub.4), filtered and concentrated in vacuo to afford a brown oil. The crude product was purified by chromatography on silica gel (80 g column, 0-10% MeOH in DCM) to afford a brown oil (100 mg), which was triturated with isohexane/ether to afford the sub-title compound (98 mg) as a brown solid.

(111) 1H NMR (DMSO-d6) 400 MHz, δ: 9.15 (s, 1H) 8.76 (s, 1H) 8.66 (d, 1H) 8.07 (d, 1H) 7.95 (d, 1H) 7.79 (d, 1H) 7.67-7.63 (m, 1H) 7.60-7.56 (m, 1H) 7.50-7.40 (m, 5H) 7.27 (d, 1H) 6.60 (s, 1H) 4.17 (t, 2H) 3.60-3.57 (m, 4H) 2.73 (t, 2H) 0.87-0.82 (m, 4H) 0.26 (s, 9H).

(112) LCMS m/z 658/660 (M+H).sup.+ (ES.sup.+)

(ii) 1-(4-((2-((7-Methyl-1H-indazol-5-yl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(4-(2-morpholinoethoxy)phenyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(113) To a stirred solution of the product from step (i) above (90 mg, 0.103 mmol) in THF/DMF (2 mL, 1:1) was added p-TSA monohydrate (39.0 mg, 0.205 mmol) followed by 7-methyl-1H-indazol-5-amine (30.2 mg, 0.205 mmol). The resulting mixture was heated at 60° C. overnight. The reaction was cooled to rt and partitioned between sodium bicarbonate solution (15 mL) and EtOAc (20 mL). The aqueous phase was extracted with EtOAc (2×20 mL). The combined organic extracts were washed with water (2×20 mL) then brine (20 mL), dried (MgSO.sub.4), filtered and concentrated in vacuo to afford a deep pink residue. The crude product was purified by chromatography on silica gel (40 g column, 0-10% MeOH in DCM) to afford a pink solid (45 mg) then purified by preparative HPLC (Waters, Acidic (0.1% Formic acid), Waters X-Select Prep-C18, 5 μm, 19×50 mm column, 20-70% MeCN in Water) to afford the title compound (14 mg) as a pink solid.

(114) 1H NMR (DMSO-d6) 400 MHz, δ: 12.85 (s, 1H) 9.46 (s, 1H) 9.21 (s, 1H) 8.81 (s, 1H) 8.38 (d, 1H) 8.11-8.04 (m, 2H) 7.82 (d, 1H) 7.63-7.50 (m, 5H) 7.43-7.30 (m, 2H) 7.17-7.13 (m, 2H) 7.02 (br s, 1H) 6.62 (s, 1H) 6.58 (d, 1H) 4.16 (t, 2H) 3.59-3.56 (m, 4H) 2.72 (t, 2H) 2.29 (s, 3H) 0.27 (s, 9H); 4H under d6-DMSO peak at 2.50 ppm.

(115) LCMS m/z 769 (M+H).sup.+ (ES.sup.+); 767 (M−H).sup.− (ES.sup.−)

Example 16

1-(3-(Ethyldimethylsilyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-((2-(phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)urea

(116) ##STR00037##

(i) (E)-2-(2-(p-Tolyl)hydrazono)acetic acid

(117) To a suspension of p-tolylhydrazine, HCl (16.66 g, 105 mmol) in water (300 mL) at rt was added conc. HCl (aq) (11.36 mL, 136 mmol) followed by dropwise addition of 50% aqueous glyoxylic acid (12.69 mL, 115 mmol) over 20 min. The mixture was stirred for a further 30 min and then the product was isolated by filtration, washing with water, then dried over MgSO.sub.4 as a solution in EtOAc (200 mL), concentrated to afford the sub-title compound (17.67 g) as an orange solid.

(118) 1H NMR (400 MHz; DMSO-d6) δ 12.30 (br. s, 1H), 11.10 (s, 1H), 7.10-7.06 (m, 3H), 7.03-6.99 (m, 2H), 2.22 (s, 3H).

(119) LCMS m/z 179 (M+H).sup.+ (ES.sup.+); 177 (M−H).sup.− (ES.sup.−)

(ii) p-Tolylcarbonohydrazonic dibromide

(120) To a stirred solution of the product from step (i) above (17.67 g, 94 mmol) in anhydrous DMF (120 mL) at 0° C. was added NBS (33.5 g, 188 mmol) portionwise over 20 min and the reaction allowed to warm to ambient temperature over 16 h. The reaction mixture was diluted with water (150 mL) and extracted with diethyl ether (3×200 mL). The combined organic extracts were washed with brine (50 mL) and dried (MgSO.sub.4). The crude product was purified by chromatography on the Companion (330 g column, 0-2.5% ether) to afford the sub-title compound (10.2 g) as a pale yellow solid.

(121) 1H NMR (400 MHz; CDCl.sub.3) δ 7.51 (br. s, 1H), 7.10-7.05 (m, 2H), 6.97-6.92 (m, 2H), 2.28 (s, 3H).

(iii) Ethyl 3-bromo-1-(p-tolyl)-1H-pyrazole-5-carboxylate

(122) To a solution of the product from step (ii) above (3.05 g, 10.45 mmol) and ethyl propiolate (5.35 mL, 52.2 mmol) in anhydrous DMF (20 mL) was added triethylamine (1.456 mL, 10.45 mmol) dropwise over 5 min with slight cooling by means of an ice-acetone bath. The ice bath was removed upon full addition and the reaction stirred for 2 h. The reaction was split between water (50 mL) and ether (150 mL). Aqueous extracted with further ether (50 mL) and the organic extracts were combined and washed with water (3×20 mL) and brine (20 mL) then dried over MgSO.sub.4, filtered and concentrated in vacuo to afford an orange solid. The crude product was purified by chromatography on the Companion (120 g column, 0-5% ether in isohexanes) to afford the sub-title compound (1.92 g) as a pale yellow, fluffy solid.

(123) 1H NMR (400 MHz; CDCl.sub.3) δ 7.30-7.22 (m, 4H), 6.99 (s, 1H), 4.24 (q, 2H), 2.41 (s, 3H), 1.26 (t, 3H).

(124) LCMS m/z 309, 311 (M+H).sup.+ (ES.sup.+)

(iv) Ethyl 3-(ethyldimethylsilyl)-1-(p-tolyl)-1H-pyrazole-5-carboxylate

(125) Tripotassium phosphate (1030 mg, 4.85 mmol), tetraethylammonium iodide (2495 mg, 9.70 mmol) and the product from step (iii) above (500 mg, 1.617 mmol) were added to a microwave tube equipped with a magnetic stirrer bar and the tube flushed with nitrogen, bis(tri-t-butylphosphine)palladium (0) (Pd-116) (71 mg, 0.139 mmol) was next added and the tube sealed and flushed with nitrogen prior to the addition of anhydrous, degassed NMP (1 mL) and ethyldimethylsilane (256 μL, 1.941 mmol). The mixture was stirred at ambient temperature for 24 h. Further catalyst was then added (30 mg) and further ethyldimethylsilane (256 μL, 1.941 mmol) and the reaction stirred for a further 24 h. After this time the reaction partitioned between water (20 mL) and ether (100 mL) and the ether layer washed with further water (20 mL), brine (20 mL), dried over MgSO.sub.4, filtered and solvent removed under reduced pressure. The crude product was purified by chromatography on the Companion (40 g column, 0-5% ether in isohexanes) to afford the sub-title compound (60 mg) as a clear, colourless oil.

(126) H NMR (400 MHz; CDCl.sub.3) δ 7.32-7.29 (m, 2H), 7.26-7.22 (m, 2H), 7.10 (s, 1H), 4.23 (q, 2H), 2.40 (s, 3H), 1.26 (t, 3H), 1.00 (t, 3H), 0.79 (q, 2H), 0.30 (s, 6H).

(127) LCMS m/z 317 (M+H).sup.+ (ES.sup.+)

(v) 3-(Ethyldimethylsilyl)-1-(p-tolyl)-1H-pyrazole-5-carboxylic acid

(128) To a solution of the product from step (iv) above (60 mg, 0.190 mmol) in ethanol (2 mL) was added 2 M aqueous NaOH (190 μL, 0.379 mmol) and the mixture stirred overnight at ambient temperature. The solvent was removed under reduced pressure and the resulting solid partitioned between EtOAc (20 mL) and 1N HCl (5 mL). The organic extract was washed with saturated brine (5 mL) and then dried over MgSO.sub.4, filtered and concentrated in vacuo to afford the sub-title compounds as a brown oil (55 mg).

(129) LCMS m/z 289 (M+H).sup.+ (ES.sup.+); 287 (M−H).sup.− (ES.sup.−)

(vi) 1-(3-(Ethyldimethylsilyl)-1-(p-tolyl)-1H-pyrazol-5-yl)-3-(4-((2-(phenylamino)pyrimidin-4-yl)oxy)naphthalen-1-yl)urea

(130) To a nitrogen purged solution of the product from step (v) above (55 mg, 0.191 mmol) in anhydrous dioxane was added DPPA (61.6 μL, 0.286 mmol) and triethylamine (80 μL, 0.572 mmol). The resulting solution was stirred at ambient temperature for 40 min prior to the addition of 4-((4-aminonaphthalen-1-yl)oxy)-N-phenylpyrimidin-2-amine (62.6 mg, 0.191 mmol). The reaction was next heated for 1 h at 100° C. The reaction mixture was concentrated to dryness to afford a brown oil and then this was purified by chromatography on the Companion (12 g column, 0-20% EtOAc in toluene) to afford the title compound (37 mg) as a pale yellow solid.

(131) 1H NMR (400 MHz; DMSO-d6) δ 9.50 (s, 1H), 9.16 (s, 1H), 8.76 (s, 1H), 8.39 (d, 1H), 8.08 (d, 1H), 7.93 (d, 1H), 7.81 (dd, 1H), 7.62 (ddd, 1H), 7.55 (ddd, 1H), 7.51-7.47 (m, 2H), 7.42-7.37 (m, 3H), 7.28 (br.d, 2H), 6.96 (t, 2H), 6.77 (t, 1H), 6.62 (s, 1H), 6.57 (d, 1H), 2.41 (s, 3H), 0.99 (t, 3H), 0.73 (q, 2H), 0.24 (s, 6H).

(132) LCMS m/z 614 (M+H).sup.+ (ES.sup.+)

Example 17

1-(2,3-Dichloro-4-((2-((7-methyl-1H-indazol-5-yl)amino)pyrimidin-4-yl)oxy)phenyl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(133) ##STR00038##

(i) 2,3-Dichloro-4-((2-chloropyrimidin-4-yl)oxy)aniline

(134) DBU (11.85 mL, 79 mmol) was added over 5 min to a stirred mixture of 4-amino-2,3-dichlorophenol (10 g, 56.2 mmol) in MeCN (150 mL) at 0-5° C. After stirring for 5 min, 2,4-dichloropyrimidine (8.95 g, 60.1 mmol) was added portionwise over 5 min then the mixture warmed to rt and stirred for 2 h. The solvent was evaporated under reduced pressure and the residue partitioned between ether (200 mL) and water (200 mL). The aqueous layer was extracted with ether (200 mL) then the combined organic layers washed with brine (200 mL), dried (MgSO.sub.4), filtered through a pad of silica and evaporated under reduced pressure. The residue was triturated with ether-isohexane, filtered and dried to afford the sub-title compound (14.4 g) as a light brown solid.

(135) 1H NMR (CDCl.sub.3) 400 MHz, δ: 8.45 (d, 1H), 6.96 (d, 1H), 6.84 (d, 1H), 6.73 (d, 1H), 4.22 (s, 2H).

(136) LCMS m/z 290/2/4 (M+H).sup.+ (ES.sup.+).

(ii) 1-(2,3-Dichloro-4-((2-chloropyrimidin-4-yl)oxy)phenyl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(137) DPPA (471 μL, 2.187 mmol) was added to a stirred solution of 1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazole-5-carboxylic acid (see, for example, Barnes, M. J. et al. Bioorg. & Med. Chem. Lett., 17(2), 354-357 (2007) and Bastian, J. A. et al. WO 2007/053394, 10 May 2007; 600 mg, 2.187 mmol) and triethylamine (762 μL, 5.47 mmol) in DMF (6 mL) under N.sub.2 at 0-5° C. After 30 min the mixture was warmed to rt and stirred for a further 1 h. The product from step (i) above (635 mg, 2.187 mmol) was added and the mixture heated at 100° C. for 2 h. The mixture was diluted with water (10 mL) and extracted with ethyl acetate (3×30 mL). The organic phases were washed with water (10 mL), brine (10 mL), dried (MgSO.sub.4) and evaporated under reduced pressure. The crude product was purified by chromatography on silica gel (40 g column, 0-40% EtOAc/iso-hexanes) to afford an off white foam. The foam was re-purified by chromatography on the Companion (80 g column, 0-30% EtOAc/toluene) to afford an off white foam. The foam was purified by chromatography on the Companion (40 g column, 30% acetone/iso-hexane) to afford the sub-title compound (512 mg) as a white glass.

(138) LCMS m/z 561, 563, 565 (M+H).sup.+ (ES.sup.+).

(iii) 1-(2,3-Dichloro-4-((2-((7-methyl-1H-indazol-5-yl)amino)pyrimidin-4-yl)oxy)phenyl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(139) A mixture of the product from step (ii) above (250 mg, 0.445 mmol), 7-methyl-1H-indazol-5-amine, HCl (123 mg, 0.667 mmol) and p-TSA monohydrate (42.3 mg, 0.222 mmol) was heated to 70° C. in DMF (2 mL) for 6 h. The mixture was diluted with water (5 mL) and saturated NaHCO.sub.3 solution (5 mL). The precipitate was collected by filtration and washed with water (5 mL) to yield a dark solid. The crude product was purified by chromatography on the Companion (40 g column, 0-100% EtOAc/iso-hexanes) to afford a solid. The solid was triturated in acetonitrile to yield the title compound (35 mg) as a white solid.

(140) 1H NMR (DMSO-d6) 400 MHz, δ: 12.94 (s, 1H), 9.61 (br s, 1H), 9.30 (s, 1H), 8.96 (s, 1H), 8.38 (d, 1H), 8.25 (d, 1H), 7.67 (br s, 1H), 7.53 (br s, 1H), 7.49-7.43 (m, 3H), 7.42-7.37 (m, 2H), 7.11 (s, 1H), 6.62 (s, 1H), 6.55 (d, 1H), 2.40 (s, 3H), 2.36 (s, 3H), 0.26 (s, 9H).

(141) LCMS m/z 672/674 (M+H)+ (ES+).

Example 18

1-(2,3-Dichloro-4-((2-((3-methoxy-5-(2-morpholinoethoxy)phenyl)amino)pyrimidin-4-yl)oxy)phenyl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(142) ##STR00039##

(143) A mixture of 1-(2,3-dichloro-4-((2-chloropyrimidin-4-yl)oxy)phenyl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea (see Example 17(ii) above; 250 mg, 0.445 mmol), 3-methoxy-5-(2-morpholino ethoxy)aniline, HCl (193 mg, 0.667 mmol) and p-TSA monohydrate (169 mg, 0.890 mmol) was heated to 70° C. in DMF (2 mL) for 6 h. The mixture was diluted with water (5 mL) and saturated NaHCO.sub.3 solution (5 mL). The precipitate was collected by filtration and washed with water (5 mL) to yield a dark purple solid. The crude product was purified by chromatography on the Companion (40 g column, 0-10% MeOH:NH.sub.4OH(9:1)/EtOAc) to afford a purple solid. The solid was purified by preparative HPLC (Waters, Acidic (0.1% Formic acid), Waters X-Select Prep-C18, 5 μm, 19×50 mm column, 5-95% MeCN in Water) to afford the title compound (86 mg) as a white solid.

(144) 1H NMR (DMSO-d6) 400 MHz, δ: 9.52 (s, 1H), 9.27 (s, 1H), 8.86 (s, 1H), 8.40 (d, 1H), 8.17 (d, 1H), 7.46-7.34 (m, 5H), 6.77-6.65 (m, 2H), 6.59 (s, 1H), 6.58 (d, 1H), 6.07 (t, 1H), 3.94 (t, 2H), 3.57 (s, 3H), 3.56-3.51 (m, 4H), 2.61 (t, 2H), 2.45-2.36 (m, 4H), 2.40 (s, 3H), 0.25 (s, 9H).

(145) LCMS m/z 778, 780 (M+H)+ (ES+); 776 (M−H)− (ES−).

Example 19

3-Methoxy-N-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-5-((4-((4-(3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-benzamide

(146) ##STR00040##

(i) 3-Methoxy-5-((4-((4-(3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)benzoic acid

(147) To a stirred solution of 1-(4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea (see Example 6(i) above; 135 mg, 0.249 mmol) in THF (6 mL) was added p-TSA monohydrate (70.9 mg, 0.373 mmol) followed by 3-amino-5-methoxybenzoic acid (62.3 mg, 0.373 mmol). The resulting mixture was heated at 60° C. overnight. The reaction was cooled to rt and the solvent removed in vacuo. The crude product was purified by chromatography on silica gel (40 g column, 0-20% MeOH/NH.sub.3 in DCM) to afford a pale pink solid. The solid was dissolved in the minimum of MeOH/DCM (1:1) and loaded on to SAX. The column was eluted with MeOH and the product eluted with 5% AcOH in MeOH. The filtrate was concentrated in vacuo to afford the sub-title product (101 mg) as a pale pink solid. The product was used in the next step without further purification.

(148) LCMS m/z 674 (M+H).sup.+ (ES.sup.+)

(ii) 3-Methoxy-N-((2S,3R,4R,5R)-2,3,4,5,6-pentahydroxyhexyl)-5-((4-((4-(3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-benzamide

(149) To a cooled solution of the product from step (i) (90 mg, 0.094 mmol) in DMF (3 mL) at 0° C. was added HATU (42.7 mg, 0.112 mmol), (2R,3R,4R,5S)-6-aminohexane-1,2,3,4,5-pentaol (20.33 mg, 0.112 mmol) and N,N-diisopropylethylamine (Hünig's Base) (49.0 μL, 0.281 mmol). The reaction mixture was stirred at it overnight. The solvent was removed in vacuo. The resulting residue was triturated with MeOH to afford a light beige solid (56 mg) at 85% purity. The crude product was purified by preparative HPLC (Waters, Acidic (0.1% formic acid), Waters X-Select Prep-C18, 5 μm, 19×50 mm column, 35-65% MeCN in water) to afford the title compound (3 mg) as a white solid.

(150) 1H NMR (DMSO-d6) 400 MHz, δ: 9.23 (s, 1H) 8.89 (s, 1H) 8.55 (s, 1H) 8.39 (d, 1H) 8.09-8.07 (m, 1H) 7.88-7.86 (m, 2H) 7.80-7.73 (m, 1H) 7.61-7.53 (m, 3H) 7.49-7.47 (m, 2H) 7.41-7.35 (m, 3H) 6.91-6.90 (m, 1H) 6.58 (s, 1H) 6.48 (d, 1H) 4.52-4.51 (m, 1H) 4.18-4.15 (m, 2H) 4.06-4.05 (m, 1H) 4.01-3.99 (m, 1H) 3.84-3.76 (m, 2H) 3.68-3.65 (m, 1H) 3.61 (s, 3H) 3.57-3.44 (m, 4H) 3.34-3.27 (m, 2H) 2.41 (s, 3H) 0.29 (s, 9H).

(151) LCMS m/z 837 (M+H).sup.+ (ES.sup.+); 835 (M−H).sup.− (ES.sup.−).

Example 20

1-(4-((2-((1-Oxoisoindolin-5-yl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(152) ##STR00041##

(153) p-TSA monohydrate (23.12 mg, 0.122 mmol) was added to a stirred solution of 1-(4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea (see Example 6(i) above; 33 mg, 0.061 mmol) and 5-aminoisoindolin-1-one (18.01 mg, 0.122 mmol) in THF/DMF (1 mL, 1:1). The resulting mixture was heated at 60° C. overnight. The reaction was cooled to rt then diluted with EtOAc (15 mL). The suspension was filtered, the product was detected in the filtrate which was subsequently concentrated in vacuo. The crude product was purified by chromatography on silica gel (12 g column, 0-10% MeOH in DCM) to afford a white solid. The crude product was purified by preparative HPLC (Waters, Acidic (0.1% Formic acid), Waters X-Select Prep-C18, 5 μm, 19×50 mm column, 35-75% MeCN in Water) to afford the title compound (7 mg) as a white solid.

(154) 1H NMR (DMSO-d6) 400 MHz, δ: 9.93 (s, 1H), 9.24 (s, 1H), 8.84 (s, 1H), 8.47 (d, 1H), 8.22 (s, 1H), 8.11 (d, 1H), 8.00 (d, 1H), 7.82-7.80 (m, 1H), 7.65-7.61 (m, 1H), 7.58-7.54 (m, 1H), 7.53-7.36 (m, 6H), 7.34-7.25 (m, 2H), 6.70 (d, 1H), 6.65 (s, 1H), 3.99 (s, 2H), 2.41 (s, 3H), 0.27 (s, 9H).

(155) LCMS m/z 655 (M+H)+ (ES+).

Example 21

1-(4-((2-((3-Methoxy-5-(2-(2-(2-methoxyethoxyl)ethoxy)ethoxy)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(156) ##STR00042##

(i) 3-Methoxy-5-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)aniline

(157) 1-Bromo-2-(2-(2-methoxyethoxyl)ethoxy)ethane (2.75 mL, 15.86 mmol) was added to a suspension of 3-amino-5-methoxyphenol (2 g, 14.37 mmol), K.sub.2CO.sub.3 (6 g, 43.4 mmol) and NaI (0.215 g, 1.437 mmol) in acetone (50 ml) and heated at reflux for 16 h. The mixture was partitioned between EtOAc (10 ml) and water (10 ml). The organic layer was separated washed with 20% w/w NaCl soln. (10 ml), dried (MgSO.sub.4), filtered and evaporated. The crude product was purified by chromatography on silica gel (80 g column, 50% EtOAc:isohexane to 100%) to afford the sub-title compound (3.2 g) as a thick brown oil.

(158) 1H NMR (400 MHz, DMSO-d6) δ 5.76-5.73 (m, 2H), 5.68 (t, 1H), 5.07 (s, 2H), 3.98-3.89 (m, 2H), 3.72-3.65 (m, 2H), 3.63 (s, 3H), 3.60-3.48 (m, 6H), 3.47-3.40 (m, 2H), 3.24 (s, 3H).

(159) LCMS m/z 286 (M+H)+ (ES+)

(ii) tert-Butyl (4-((2-((3-methoxy-5-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)carbamate

(160) tert-Butyl (4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)carbamate (see, for example, Ito, K. et al., WO 2010/067130, 17 Jun. 2010; 1 g, 2.69 mmol), the product from step (i) above (1.15 g, 4.03 mmol) and p-TSA monohydrate (0.102 g, 0.538 mmol) in DMF (10 ml) was heated at 65° C. (block temperature) for 8 h. The mixture was cooled and partitioned between EtOAc (150 ml) and aq sat NaHCO.sub.3 (50 ml). The organic layer was washed with water (50 ml), dried (MgSO.sub.4), filtered and evaporated under reduced pressure. The crude product was purified by chromatography on silica gel (40 g column, 0-5% MeOH/DCM) then further purified by chromatography on silica gel (80 g column, 0-100% EtOAc/isohexane) to afford the sub-title compound (1.01 g) as a foam.

(161) LCMS m/z 621 (M+H).sup.+ (ES.sup.+)

(iii) 4-((4-Aminonaphthalen-1-yl)oxy)-N-(3-methoxy-5-(2-(2-(2-methoxyethoxy)ethoxy)-ethoxy)phenyl)pyrimidin-2-amine

(162) TFA (3 mL, 38.9 mmol) was added to a stirred solution of the product from step (ii) above (1 g, 1.611 mmol) in DCM (12 ml) at rt. The mixture was stirred for 2 h then evaporated under reduced pressure. The residue was partitioned between DCM (100 ml) and sat aq NaHCO.sub.3 soln (50 ml), the organic layer separated, washed with water (50 ml), dried (MgSO.sub.4), filtered and evaporated under reduced pressure. MeOH (10 ml) was added and the residue evaporated to afford the sub-title compound (840 mg) as a solid.

(163) 1H NMR (400 MHz; DMSO-d6) δ 9.42 (s, 1H), 8.33 (d, 1H), 8.14-8.12 (m, 1H), 7.64-7.62 (m, 1H), 7.46-7.41 (m, 2H), 7.11 (d, 1H), 6.87 (br s, 2H), 6.68 (d, 1H), 6.34 (d, 1H), 6.04 (s, 1H), 5.79 (s, 2H), 3.87-3.85 (m, 2H), 3.68-3.66 (m, 2H), 3.56-3.50 (m, 9H), 3.43-3.41 (m, 2H), 3.22 (s, 3H).

(164) LCMS m/z 521 (M+H).sup.+ (ES.sup.+)

(iv) 1-(4-((2-((3-Methoxy-5-(2-(2-(2-methoxyethoxy)ethoxy)ethoxy)phenyl)amino)-pyrimidin-4-yl)oxy)naphthalen-1-yl)-3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)urea

(165) DPPA (80 μL, 0.360 mmol) was added to a stirred solution of 1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazole-5-carboxylic acid (see, for example, Barnes, M. J. et al. Bioorg. & Med. Chem. Lett., 17(2), 354-357 (2007) and Bastian, J. A. et al. WO 2007/053394, 10 May 2007; 100 mg, 0.364 mmol) and Et.sub.3N (130 μL, 0.933 mmol) in DMF (2 mL) at 0-5° C. After 30 min the mixture was warmed to rt and stirred for a further 1 h. the product from step (iii) above (190 mg, 0.364 mmol) was added and the mixture heated at 100° C. for 1 h then cooled and partitioned between EtOAc (100 ml) and water (50 ml). The organic layer was separated, washed with water (50 ml), 20% w/w brine (50 ml), dried (MgSO.sub.4), filtered and evaporated under reduced pressure to a brown gum. The crude product was purified by chromatography on the Companion (40 g column, 50% EtOAc:isohexane to 100%) then triturated with Et.sub.2O (2 mL) to afford the title compound (88 mg) as a colourless solid.

(166) 1H NMR (400 MHz, DMSO-d6) δ 9.43 (s, 1H), 9.12 (s, 1H), 8.79 (s, 1H), 8.40 (d, 1H), 8.06 (d, 1H), 7.95 (d, 1H), 7.87-7.75 (m, 1H), 7.67-7.52 (m, 2H), 7.51-7.44 (m, 2H), 7.44-7.34 (m, 3H), 6.91-6.70 (m, 2H), 6.62 (s, 1H), 6.53 (d, 1H), 6.03 (t, 1H), 3.85 (t, 2H), 3.70-3.59 (m, 2H), 3.57-3.44 (m, 9H), 3.44-3.37 (m, 2H), 3.21 (s, 3H), 2.41 (s, 3H), 0.26 (s, 9H).

(167) LCMS m/z 792 (M+H)+ (ES+)

Example 22

3-Ethynyl-N-(2-morpholinoethyl)-5-((4-((4-(3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)benzamide

(168) ##STR00043##

(i) 3-Amino-5-bromo-N-(2-morpholinoethyl)benzamide

(169) 2-Morpholinoethanamine (0.911 mL, 6.94 mmol) was added to an ice cold suspension of T3P (1-propanephosphonic acid cyclic anhydride) (2.76 mL, 4.63 mmol), 3-amino-5-bromobenzoic acid (1 g, 4.63 mmol) and TEA (1.936 mL, 13.89 mmol) in DCM (20 ml). Allowed to warm to room temperature and stirred overnight. More T3P (2.76 mL, 4.63 mmol) and 2-morpholinoethanamine (0.911 mL, 6.94 mmol) were added and stirred for a further 1 h. Partitioned with sat. NaHCO.sub.3 soln. (20 ml), the aqueous layer separated and partitioned with fresh DCM (20 ml). The organics separated, bulked and partitioned with 20% w/w NaCl soln. The organic layer was separated, dried (MgSO.sub.4) filtered and solvent evaporated. The crude product was purified by chromatography on the Companion (40 g column, 2% MeOH:DCM to 5%) to afford the sub-title compound (1.4 g) as a yellow crystalline solid.

(170) 1H NMR (400 MHz, DMSO-d6) δ 8.28 (t, 1H), 7.06 (t, 1H), 6.98 (dd, 1H), 6.85 (t, 1H), 5.58 (s, 2H), 3.57 (t, 4H), 3.33 (m, 2H), 2.41 (m, 6H).

(171) LCMS m/z 328/330 (M+H).sup.+ (ES.sup.+)

(ii) 3-Amino-N-(2-morpholinoethyl)-5-((triisopropylsilyl)ethynyl)benzamide

(172) Pd(PPh.sub.3).sub.4 (176 mg, 0.152 mmol) was added to a degassed suspension of the product from step (i) above (500 mg, 1.523 mmol), copper(I) iodide (29.0 mg, 0.152 mmol), and ethynyltriisopropylsilane (0.513 mL, 2.285 mmol) in TEA (3 mL) and DMF (3 mL), heated at 80° C. (block temp.) for 1 h then cooled, filtered on Celite and solvents evaporated. The crude product was purified by chromatography on the Companion (12 g column, 5% MeOH:DCM to 10%) to afford the sub-title compound (600 mg) as a pale yellow gum.

(173) 1H NMR (400 MHz, CDCl.sub.3) δ 11.05 (s, 1H), 7.16 (t, 1H), 7.13 (t, 1H), 6.90 (dd, 1H), 3.83 (s, 2H), 3.77 (t, 4H), 3.56 (q, 2H), 2.65 (s, 2H), 2.57 (s, 4H), 1.13 (s, 21H).

(174) LCMS m/z 430 (M+H).sup.+ (ES.sup.+)

(iii) 3-Amino-5-ethynyl-N-(2-morpholinoethyl)benzamide

(175) The product from step (ii) above (500 mg, 1.164 mmol) was dissolved in THF (5 mL) and TBAF (1M in THF, 1164 μL, 1.164 mmol) added and stirred for 1 h. TBAF (1164 μL, 1.164 mmol) added again and stirred for 30 min. Reaction partitioned between water (10 ml) and ethyl acetate (10 ml), organic layer separated and washed with 20% w/w NaCl soln. Organic layer was separated, dried (MgSO.sub.4) filtered and evaporated. The crude product was purified by chromatography on the Companion (12 g column, 2% MeOH:DCM to 5%) to afford the sub-title compound (260 mg) as a colourless gum.

(176) 1H NMR (400 MHz, CDCl.sub.3) δ 7.15 (m, 2H), 6.91 (dd, 1H), 6.67 (s, 1H), 3.85 (s, 2H), 3.74 (t, 4H), 3.53 (q, 2H), 3.07 (s, 1H), 2.59 (t, 2H), 2.51 (t, 4H).

(177) LCMS m/z 274 (M+H).sup.+ (ES.sup.+)

(iv) tert-Butyl (4-((2-((3-ethynyl-5-(2-morpholinoethyl)carbamoyl)phenyl)amino)pyrimidin-4-yl)oxy)naphthalen-1-yl)carbamate

(178) tert-Butyl (4-((2-chloropyrimidin-4-yl)oxy)naphthalen-1-yl)carbamate (see, for example, Ito, K. et al., WO 2010/067130, 17 Jun. 2010; 6.46 g, 17.37 mmol), the product from step (iii) above (7.12 g, 26.0 mmol) and p-TSA monohydrate (5.62 g, 29.5 mmol) in DMF (60 ml) was heated at 60° C. (block temperature, 55° C. internal temperature) for 7 h. The mixture was cooled and added dropwise to sat. aq NaHCO.sub.3 (1 L). The solid was filtered, washed with water (50 ml) then isohexane (100 ml). The amorphous solid was stirred in MeOH (200 ml) and product crystallised. Slurried overnight, then filtered and solid washed with MeOH (20 ml) and dried to afford the sub-title compound (8 g).

(179) 1H NMR (400 MHz, DMSO-d6) δ 9.76 (s, 1H), 9.32 (s, 1H), 8.45 (d, 1H), 8.41-8.33 (m, 1H), 8.16-8.03 (m, 2H), 7.90 (t, 1H), 7.85-7.78 (m, 1H), 7.67-7.51 (m, 3H), 7.48-7.37 (m, 2H), 6.58 (d, 1H), 4.16 (s, 1H), 3.56 (t, 4H), 3.46-3.27 (m, 2H), 2.49-2.30 (m, 6H), 1.52 (s, 9H). 10% w/w de-BOC compound.

(180) LCMS m/z 609 (M+H).sup.+ (ES.sup.+)

(v) 3-((4-((4-Aminonaphthalen-1-yl)oxy)pyrimidin-2-yl)amino)-5-ethynyl-N-(2-morpholinoethyl)benzamide

(181) TFA (22 mL, 286 mmol) was added dropwise to a stirred solution of the product from step (iv) above (9 g, 14.05 mmol) in DCM (50 mL). The reaction was stirred at rt for 2 h, then added dropwise to stirred water (100 mL) and 1M potassium carbonate solution (280 mL, 280 mmol) and stirring continued until effervescence ceased. The mixture was extracted with DCM (2×250 mL) then the combined organic phases were dried (MgSO.sub.4) and concentrated under reduced pressure. The crude product was purified by chromatography on the Companion (120 g column, 2% MeOH:DCM to 6%) to afford the sub-title compound (6.7 g) as a pale brown foam.

(182) 1H NMR (400 MHz, DMSO-d6) δ 9.77 (s, 1H), 8.39 (t, 1H), 8.36 (d, 1H), 8.17-8.10 (m, 1H), 8.06 (s, 1H), 7.94 (dd, 1H), 7.67-7.59 (m, 1H), 7.49-7.38 (m, 3H), 7.15 (d, 1H), 6.70 (d, 1H), 6.37 (d, 1H), 5.79 (s, 2H), 4.20 (s, 1H), 3.56 (t, 4H), 3.41-3.30 (m, 2H), 2.48-2.34 (m, 6H).

(183) LCMS m/z 509 (M+H).sup.+ (ES.sup.+)

(vi) 3-Ethynyl-N-(2-morpholinoethyl)-5-((4-((4-(3-(1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazol-5-yl)ureido)naphthalen-1-yl)oxy)pyrimidin-2-yl)amino)benzamide

(184) To a stirred solution of 1-(p-tolyl)-3-(trimethylsilyl)-1H-pyrazole-5-carboxylic acid (see, for example, Barnes, M. J. et al. Bioorg. & Med. Chem. Lett., 17(2), 354-357 (2007) and Bastian, J. A. et al. WO 2007/053394, 10 May 2007; 150 mg, 0.547 mmol) and triethylamine (200 μL, 1.435 mmol) in DMF (2 mL) under N.sub.2 at 0° C. was added DPPA (180 μL, 0.835 mmol). The mixture was stirred at rt for 20 minutes before adding the product from step (v) above (278 mg, 0.547 mmol) and heating to 100° C. for 1 h. Upon cooling, water (10 mL) was added dropwise and the suspension left stirring overnight. The resulting solid was collected by filtration washing with water. The crude product was purified by chromatography on the Companion (40 g column, 1-10% MeOH in DCM) to afford the product as a dark yellow glassy solid. The solid was suspended in a mixture of EtOAc and Et.sub.2O (1:4) and sonicated for 20 minutes by which time a gel-like solid was observed. The solid was collected by filtration washing with more EtOAc to afford the title compound (66 mg) as a pale pink solid.

(185) 1H NMR (DMSO-d6) 400 MHz, δ: 9.76 (s, 1H), 9.11 (s, 1H), 8.78 (s, 1H), 8.44 (d, 1H), 8.36 (bs, 1H), 8.05-8.07 (m, 2H), 7.94 (d, 1H), 7.86 (s, 1H), 7.82 (d, 1H), 7.55-7.65 (m, 2H), 7.49 (d, 2H), 7.40-7.44 (m, 4H), 6.62 (s, 1H), 6.57 (d, 1H), 4.12 (s, 1H), 3.56 (t, 4H), 2H under water peak, 2.34-2.45 (m, 6H), 2.42 (s, 3H), 0.27 (s, 9H).

(186) LCMS m/z 391 (M+2H).sup.2+ (ES+)

(187) Biological Testing: Experimental Methods

(188) Enzyme Inhibition Assays

(189) The enzyme inhibitory activities of compounds disclosed herein are determined by FRET using synthetic peptides labelled with both donor and acceptor fluorophores (Z-LYTE, Invitrogen Ltd., Paisley, UK).

(190) p38 MAPKα Enzyme Inhibition

(191) The following two assay variants can be used for determination of p38 MAPKα inhibition.

(192) Method 1

(193) The inhibitory activities of test compounds against the p38 MAPKα isoform (MAPK14: Invitrogen), are evaluated indirectly by determining the level of activation/phosphorylation of the down-stream molecule, MAPKAP-K2. The p38 MAPKα protein (80 ng/mL, 2.5 μL) is mixed with the test compound (2.5 μL of either 4 μg/mL, 0.4 μg/mL, 0.04 μg/mL or 0.004 μg/mL) for 2 hr at RT. The mix solution (2.5 μL) of the p38α inactive target MAPKAP-K2 (Invitrogen, 600 ng/mL) and FRET peptide (8 μM; a phosphorylation target for MAPKAP-K2) is then added and the kinase reaction is initiated by adding ATP (40 μM, 2.5 μL). The mixture is incubated for 1 hr at RT. Development reagent (protease, 5 μL) is added for 1 hr prior to detection in a fluorescence microplate reader (Varioskan® Flash, ThermoFisher Scientific).

(194) Method 2

(195) This method follows the same steps as Method 1 above, but utilises a higher concentration of the p38 MAPKα protein (2.5 μL of 200 ng/mL protein instead of 2.5 μL of 80 ng/mL protein) for mixing with the test compound.

(196) p38 MAPKγ Enzyme Inhibition

(197) The inhibitory activities of compounds of the invention against p38MAPKγ (MAPK12: Invitrogen), are evaluated in a similar fashion to that described hereinabove. The enzyme (800 ng/mL, 2.5 μL) is incubated with the test compound (2.5 μL at either 4 μg/mL, 0.4 μg/mL, 0.04 μg/mL, or 0.004 μg/mL) for 2 hr at RT. The FRET peptides (8 μM, 2.5 μL), and appropriate ATP solution (2.5 μL, 400 μM) is then added to the enzymes/compound mixtures and incubated for 1 hr. Development reagent (protease, 5 μL) is added for 1 hr prior to detection in a fluorescence microplate reader (Varioskan® Flash, Thermo Scientific).

(198) c-Src and Syk Enzyme Inhibition

(199) The inhibitory activities of compounds of the invention against c-Src and Syk enzymes (Invitrogen), are evaluated in a similar fashion to that described hereinabove. The relevant enzyme (3000 ng/mL or 2000 ng/mL respectively, 2.5 μL) is incubated with the test compound (either 4 μg/mL, 0.4 μg/mL, 0.04 μg/mL, or 0.004 μg/mL, 2.5 μL each) for 2 hr at RT. The FRET peptides (8 μM, 2.5 μL), and appropriate ATP solutions (2.5 μL, 800 μM for c-Src, and 60 μM ATP for Syk) are then added to the enzymes/compound mixtures and incubated for 1 hr. Development reagent (protease, 5 μL) is added for 1 hr prior to detection in a fluorescence microplate reader (Varioskan® Flash, ThermoFisher Scientific).

(200) GSK 3α Enzyme Inhibition

(201) The following two assay variants can be used for determination of GSK 3α inhibition.

(202) Method 1

(203) The inhibitory activities of compounds of the invention against the GSK 3α enzyme isoform (Invitrogen), are evaluated by determining the level of activation/phosphorylation of the target peptide. The GSK3-α protein (500 ng/mL, 2.5 μL) is mixed with the test compound (2.5 μL at either 4 μg/mL, 0.4 μg/mL, 0.04 μg/mL, or 0.004 μg/mL) for 2 hr at RT. The FRET peptide (8 μM, 2.5 μL), which is a phosphorylation target for GSK3α, and ATP (40 μM, 2.5 μL) are then added to the enzyme/compound mixture and the resulting mixture incubated for 1 hr. Development reagent (protease, 5 μL) is added for 1 hr prior to detection in a fluorescence microplate reader (Varioskan® Flash, ThermoFisher Scientific).

(204) In all cases, the site-specific protease cleaves non-phosphorylated peptide only and eliminates the FRET signal. Phosphorylation levels of each reaction are calculated using the ratio of coumarin emission (donor) over fluorescein emission (acceptor), for which high ratios indicate high phosphorylation and low ratios indicate low phosphorylation levels. The percentage inhibition of each reaction is calculated relative to non-inhibited control and the 50% inhibitory concentration (IC.sub.50 value) is then calculated from the concentration-response curve.

(205) Method 2

(206) This method follows the same steps as Method 1 above, but utilises a shorter period of mixing of the test compound (105 minutes instead of 2 hours) with the GSK3-α protein.

(207) Cellular Assays

(208) The compounds of the invention were studied using one or more of the following assays.

(209) (a) LPS-Induced TNFα/IL-8 Release in d-U937 Cells

(210) U937 cells, a human monocytic cell line, are differentiated to macrophage-type cells by incubation with phorbol myristate acetate (PMA; 100 ng/mL) for 48 to 72 hr. Cells are pre-incubated with final concentrations of test compound for 2 hr and are then stimulated with 0.1 μg/mL of LPS (from E. Coli: O111:B4, Sigma) for 4 hr. The supernatant is collected for determination of TNFα and IL-8 concentrations by sandwich ELISA (Duo-set, R&D systems). The inhibition of TNFα production is calculated as a percentage of that achieved by 10 μg/mL of BIRB796 at each concentration of test compound by comparison against vehicle control. The relative 50% effective concentration (REC.sub.50) is determined from the resultant concentration-response curve. The inhibition of IL-8 production is calculated at each concentration of test compound by comparison with vehicle control. The 50% inhibitory concentration (IC.sub.50) is determined from the resultant concentration-response curve.

(211) (b) LPS-Induced TNFα/IL-8 Release in PBMC Cells

(212) Peripheral blood mononuclear cells (PBMCs) from healthy subjects are separated from whole blood using a density gradient (Lymphoprep, Axis-Shield Healthcare). The PBMCs are seeded in 96 well plates and treated with compounds at the desired concentration for 2 hours before addition of 1 ng/mL LPS (Escherichia Coli 0111:B4 from Sigma Aldrich) for 24 hours under normal tissue culture conditions (37° C., 5% CO.sub.2). The supernatant is harvested for determination of IL-8 and TNFα concentrations by sandwich ELISA (Duo-set, R&D systems) and read on the fluorescence microplate reader (Varioskan® Flash, ThermoFisher Scientific). The concentration at 50% inhibition (IC.sub.50) of IL-8 and TNFα production is calculated from the dose response curve.

(213) (c) IL-2 and IFN Gamma Release in CD3/CD28 Stimulated PBMC Cells

(214) PBMCs from healthy subjects are separated from whole blood using a density gradient (Lymphoprep, Axis-Shield Healthcare). Cells are added to a 96 well plate pre-coated with a mixture of CD3/CD38 monoclonal antibodies (0.3 μg/mL eBioscience and 3 μg/mL BD Pharmingen respectively). Compound at the desired concentration is then added to the wells and the plate left for 3 days under normal tissue culture conditions. Supernatants are harvested and IL-2 and IFN gamma release determined by Sandwich ELISA (Duo-set, R&D System). The IC.sub.50 is determined from the dose response curve.

(215) (d) IL-1β-Induced IL-8 Release in HT29 Cells

(216) HT29 cells, a human colon adenocarcinoma cell line, are plated in a 96 well plate (24 hrs) and pre-treated with compounds at the desired concentration for 2 hours before addition of 5 ng/mL of IL-1β (Abcam) for 24 hours. Supernatants are harvested for IL-8 quantification by Sandwich ELISA (Duo-set, R&D System). The IC.sub.50 is determined from the dose response curve.

(217) (e) LPS-Induced IL-8 and TNFα Release in Primary Macrophages

(218) PBMCs from healthy subjects are separated from whole blood using a density gradient (Lymphoprep, Axis-Shield Healthcare). Cells are incubated for 2 hrs and non-adherent cells removed by washing. To differentiate the cells to macrophages the cells are incubated with 5 ng/mL of GM-CSF (Peprotech) for 7 days under normal tissue culture conditions. Compounds are then added to the cells at the desired concentration for a 2 hour pre-treatment before stimulation with 10 ng/mL LPS for 24 hours. Supernatants are harvested and IL-8 and TNFα release determined by Sandwich ELISA (Duo-set, R&D System). The IC.sub.50 is determined from the dose response curve.

(219) (f) Poly I:C-Induced ICAM-1 Expression in BEAS2B Cells

(220) Poly I:C is used in these studies as a simple, RNA virus mimic. Poly I:C-Oligofectamine mixture (1 μg/mL Poly I:C, ±2% Oligofectamine, 25 μL; Invivogen Ltd., San Diego, Calif., and Invitrogen, Carlsbad, Calif., respectively) is transfected into BEAS2B cells (human bronchial epithelial cells, ATCC). Cells are pre-incubated with final concentrations of test compounds for 2 hr and the level of ICAM1 expression on the cell surface is determined by cell-based ELISA. At a time point 18 hr after poly I:C transfection, cells are fixed with 4% formaldehyde in PBS and then endogenous peroxidase is quenched by the addition of washing buffer (100 μL, 0.05% Tween in PBS: PBS-Tween) containing 0.1% sodium azide and 1% hydrogen peroxide. Cells are washed with wash-buffer (3×200 μL) and after blocking the wells with 5% milk in PBS-Tween (100 μL) for 1 hr, the cells are incubated with anti-human ICAM-1 antibody (50 μL; Cell Signalling Technology, Danvers, Mass.) in 1% BSA PBS overnight at 4° C.

(221) The cells are washed with PBS-Tween (3×200 μL) and incubated with the secondary antibody (100 μL; HRP-conjugated anti-rabbit IgG, Dako Ltd., Glostrup, Denmark). The cells are then incubated with of substrate (50 μL) for 2-20 min, followed by the addition of stop solution (50 μL, 1N H.sub.2SO.sub.4). The ICAM-1 signal is detected by reading the absorbance at 450 nm against a reference wavelength of 655 nm using a spectrophotometer. The cells are then washed with PBS-Tween (3×200 μL) and total cell numbers in each well are determined by reading absorbance at 595 nm after Crystal Violet staining (50 μL of a 2% solution in PBS) and elution by 1% SDS solution (100 μL) in distilled water. The measured OD 450-655 readings are corrected for cell number by dividing with the OD595 reading in each well. The inhibition of ICAM-1 expression is calculated at each concentration of test compound by comparison with vehicle control. The 50% inhibitory concentration (IC.sub.50) is determined from the resultant concentration-response curve.

(222) (g) Cell Mitosis Assay

(223) Peripheral blood mononucleocytes (PBMCs) from healthy subjects are separated from whole blood (Quintiles, London, UK) using a density gradient (Histopaque®-1077, Sigma-Aldrich, Poole, UK). The PBMCs (3 million cells per sample) are subsequently treated with 2% PHA (phytohaemagglutinin, Sigma-Aldrich, Poole, UK) for 48 hr, followed by a 20 hr exposure to varying concentrations of test compounds. At 2 hr before collection, PBMCs are treated with demecolcine (0.1 μg/mL; Invitrogen, Paisley, UK) to arrest cells in metaphase. To observe mitotic cells, PBMCs are permeabilised and fixed by adding Intraprep (50 μL; Beckman Coulter, France), and stained with anti-phospho-histone 3 (0.26 ng/L; #9701; Cell Signalling, Danvers, Mass.) and propidium iodide (1 mg/mL; Sigma-Aldrich, Poole, UK) as previously described (Muehlbauer P. A. and Schuler M. J., Mutation Research, 2003, 537:117-130). Fluorescence is observed using an ATTUNE flow cytometer (Invitrogen, Paisley, UK), gating for lymphocytes. The percentage inhibition of mitosis is calculated for each treatment relative to vehicle (0.5% DMSO) treatment.

(224) (h) Rhinovirus-Induced IL-8 Release and ICAM-1 Expression

(225) Human rhinovirus RV16 is obtained from the American Type Culture Collection (Manassas, Va.). Viral stocks are generated by infecting Hela cells with HRV until 80% of the cells are cytopathic.

(226) BEAS2B cells are infected with HRV at an MOI of 5 and incubated for 2 hr at 33° C. with gentle shaking for to promote absorption. The cells are then washed with PBS, fresh media added and the cells are incubated for a further 72 hr. The supernatant is collected for assay of IL-8 concentrations using a Duoset ELISA development kit (R&D systems, Minneapolis, Minn.).

(227) The level of ICAM1 expressing cell surface is determined by cell-based ELISA. At 72 hr after infection, cells are fixed with 4% formaldehyde in PBS. After quenching endogenous peroxidase by adding 0.1% sodium azide and 1% hydrogen peroxide, wells are washed with wash-buffer (0.05% Tween in PBS: PBS-Tween). After blocking well with 5% milk in PBS-Tween for 1 hr, the cells are incubated with anti-human ICAM-1 antibody in 5% BSA PBS-Tween (1:500) overnight. Wells are washed with PBS-Tween and incubated with the secondary antibody (HRP-conjugated anti-rabbit IgG, Dako Ltd.). The ICAM-1 signal is detected by adding substrate and reading at 450 nm with a reference wavelength of 655 nm using a spectrophotometer. The wells are then washed with PBS-Tween and total cell numbers in each well are determined by reading absorbance at 595 nm after Crystal Violet staining and elution by 1% SDS solution. The measured OD.sub.450-655 readings are corrected for cell number by dividing with the OD.sub.595 reading in each well. Compounds are added 2 hr before HRV infection and 2 hr after infection when non-infected HRV is washed out.

(228) (i) Assessment of HRV16 Induced CPE in MRC5

(229) MRC-5 cells are infected with HRV16 at an MOI of 1 in DMEM containing 5% FCS and 1.5 mM MgCl.sub.2, followed by incubation for 1 hr at 33° C. to promote adsorption. The supernatants are aspirated, and then fresh media added followed by incubation for 4 days. Where appropriate, cells are pre-incubated with compound or DMSO for 2 hr, and the compounds and DMSO added again after washout of the virus.

(230) Supernatants are aspirated and incubated with methylene blue solution (100 μL, 2% formaldehyde, 10% methanol and 0.175% Methylene Blue) for 2 hr at RT. After washing, 1% SDS in distilled water (100 μL) is added to each well, and the plates are shaken lightly for 1-2 hr prior to reading the absorbance at 660 nm. The percentage inhibition for each well is calculated. The IC.sub.50 value is calculated from the concentration-response curve generated by the serial dilutions of the test compounds.

(231) (j) In Vitro RSV Virus Load in Primary Bronchial Epithelial Cells

(232) Normal human bronchial epithelial cells (NHBEC) grown in 96 well plates are infected with RSV A2 (Strain A2, HPA, Salisbury, UK) at an MOI of 0.001 in the LHC8 Media:RPMI-1640 (50:50) containing 15 mM magnesium chloride and incubated for 1 hr at 37° C. for adsorption. The cells are then washed with PBS (3×200 μL), fresh media (200 μL) is added and incubation continued for 4 days. Where appropriate, cells are pre-incubated with the compound or DMSO for 2 hr, and then added again after washout of the virus.

(233) The cells are fixed with 4% formaldehyde in PBS solution (50 μL) for 20 min, washed with WB (3×200 μL), (washing buffer, PBS including 0.5% BSA and 0.05% Tween-20) and incubated with blocking solution (5% condensed milk in PBS) for 1 hr. Cells are then washed with WB (3×200 μL) and incubated for 1 hr at RT with anti-RSV (2F7) F-fusion protein antibody (40 μL; mouse monoclonal, lot 798760, Cat. No. ab43812, Abcam) in 5% BSA in PBS-tween. After washing, cells are incubated with an HRP-conjugated secondary antibody solution (50 μL) in 5% BSA in PBS-Tween (lot 00053170, Cat. No. P0447, Dako) and then TMB substrate added (50 μL; substrate reagent pack, lot 269472, Cat. No. DY999, R&D Systems, Inc.). This reaction is stopped by the addition of 2N H.sub.2SO.sub.4 (50 μL) and the resultant signal is determined colourimetrically (OD: 450 nm with a reference wavelength of 655 nm) in a microplate reader (Varioskan® Flash, ThermoFisher Scientific).

(234) Cells are then washed and a 2.5% crystal violet solution (50 μL; lot 8656, Cat. No. PL7000, Pro-Lab Diagnostics) is applied for 30 min. After washing with WB, 1% SDS in distilled water (100 μL) is added to each well, and plates are shaken lightly on the shaker for 1 hr prior to reading the absorbance at 595 nm. The measured OD.sub.450-655 readings are corrected to the cell number by dividing the OD.sub.450-655 by the OD.sub.595 readings. The percentage inhibition for each well is calculated and the IC.sub.50 value is calculated from the concentration-response curve generated from the serial dilutions of compound.

(235) (k) Cell Viability Assay: MTT Assay

(236) Differentiated U937 cells are pre-incubated with each test compound (final concentration 1 μg/mL or 10 μg/mL in 200 μL media indicated below) under two protocols: the first for 4 hr in 5% FCS RPMI1640 media and the second in 10% FCS RPMI1640 media for 24 h. The supernatant is replaced with new media (200 μL) and MTT stock solution (10 μL, 5 mg/mL) is added to each well. After incubation for 1 hr the media are removed, DMSO (200 μL) is added to each well and the plates are shaken lightly for 1 hr prior to reading the absorbance at 550 nm. The percentage loss of cell viability is calculated for each well relative to vehicle (0.5% DMSO) treatment. Consequently an apparent increase in cell viability for drug treatment relative to vehicle is tabulated as a negative percentage.

(237) (l) Human Biopsy Assay

(238) Intestinal mucosa biopsies are obtained from the inflamed regions of the colon of IBD patients. The biopsy material is cut into small pieces (2-3 mm) and placed on steel grids in an organ culture chamber at 37° C. in a 5% CO.sub.2/95% O.sub.2 atmosphere in serum-free media. DMSO control or test compounds at the desired concentration are added to the tissue and incubated for 24 hr in the organ culture chamber. The supernatant is harvested for determination of IL-6, IL-8, IL-1β and TNFα levels by R&D ELISA. Percentage inhibition of cytokine release by the test compounds is calculated relative to the cytokine release determined for the DMSO control (100%).

(239) (m) Accumulation of β Catenin in d-U937 Cells

(240) U937 cells, a human monocytic cell line, are differentiated into macrophage-type cells by incubation with PMA; (100 ng/mL) for between 48 to 72 hr. The cells are then incubated with either final concentrations of test compound or vehicle for 18 hr. The induction of β-catenin by the test compounds is stopped by replacing the media with 4% formaldehyde solution. Endogenous peroxide activity is neutralised by incubating with quenching buffer (100 μL, 0.1% sodium azide, 1% H.sub.2O.sub.2 in PBS with 0.05% Tween-20) for 20 min. The cells are washed with washing buffer (200 μL; PBS containing 0.05% Tween-20) and incubated with blocking solution (200 μL; 5% milk in PBS) for 1 hr, re-washed with washing buffer (200 μL) and then incubated overnight with anti-β-catenin antibody solution (50 μL) in 1% BSA/PBS (BD, Oxford, UK).

(241) After washing with washing buffer (3×200 μL; PBS containing 0.05% Tween-20), cells are incubated with an HRP-conjugated secondary antibody solution (100 μL) in 1% BSA/PBS (Dako, Cambridge, UK) and the resultant signal is determined colourimetrically (OD: 450 nm with a reference wavelength of 655 nm) using TMB substrate (50 μL; R&D Systems, Abingdon, UK). This reaction is stopped by addition of 1N H.sub.2SO.sub.4 solution (50 μL). Cells are then washed with washing buffer and 2% crystal violet solution (50 μL) is applied for 30 min. After washing with washing buffer (3×200 μL), 1% SDS (100 μL) is added to each well and the plates are shaken lightly for 1 hr prior to measuring the absorbance at 595 nm (Varioskan® Flash, Thermo-Fisher Scientific).

(242) The measured OD.sub.450-655 readings are corrected for cell number by dividing the OD.sub.450-655 by the OD.sub.595 readings. The percentage induction for each well is calculated relative to vehicle, and the ratio of induction normalised in comparison with the induction produced by a standard control comprising of N-(4-(4-(3-(3-tert-butyl-1-p-tolyl-1H-pyrazol-5-yl)ureido)naphthalen-1-yloxy)pyridin-2-yl)-2-methoxyacetamide (1 μg/m L) which is defined as unity. A signal less than 0.15 of that observed for the standard control is designated as “-ve”.

(243) (n) T Cell Proliferation

(244) PBMCs from healthy subjects are separated from whole blood using a density gradient (Lymphoprep, Axis-Shield Healthcare). The lymphocyte fraction is first enriched for CD4+ T cells by negative magnetic cell sorting as per the manufacturer's instructions (Miltenyi Biotec 130-091-155). Naïve CD4+ T cells are then separated using positive magnetic selection of CD45RA+ cells using microbeads as per the manufacturer's instructions (130-045-901). Cells are plated at 2×10.sup.5 cells per well in 100 μL RPMI/10% FBS on 96 well flat bottomed plate (Corning Costar). 25 μL of test compound are diluted to the appropriate concentration (8× final conc.) in normal medium and added to duplicate wells on the plate to achieve a dose response range of 0.03 ng/mL-250 ng/mL. DMSO is added as a negative control. Plates are allowed to pre-incubate for 2 hours before stimulation with 1 μg/mL anti-CD3 (OKT3; eBioscience). After 72 h, the medium in each well is replaced with 150 μL of fresh medium containing 10 μM BrdU (Roche). After 16 h, the supernatant is removed, the plate is dried and the cells fixed by adding 100 μL of fix/denature solution to each well for 20 min as per the manufacturer's instructions (Roche). Plates are washed once with PBS before addition of the anti-BrdU detection antibody and incubated for 90 mins at room temperature. Plates are then washed gently 3× with the wash buffer supplied and developed by addition of 100 μL of substrate solution. The reaction is stopped by addition of 50 μL of 1 M H.sub.2SO.sub.4, and read for absorbance at 450 nm on a plate reader (Varioskan® Flash, ThermoFisher Scientific). The IC.sub.50 is determined from the dose response curve.

(245) (o) IL-2 and IFNγ Release in CD3/CD28 Stimulated LPMC Cells from IBD Patients

(246) Lamina propria mononuclear cells (LPMCs) are isolated and purified from inflamed IBD mucosa of surgical specimens or from normal mucosa of surgical specimens as follows: The mucosa is removed from the deeper layers of the surgical specimens with a scalpel and cut in fragments 3-4 mm size. The epithelium is removed by washing the tissue fragments three times with 1 mM EDTA (Sigma-Aldrich, Poole, UK) in HBSS (Sigma-Aldrich) with agitation using a magnetic stirrer, discarding the supernatant after each wash. The sample is subsequently treated with type 1A collagenase (1 mg/mL; Sigma-Aldrich) for 1 h with stirring at 37° C. The resulting cell suspension is then filtered using a 100 μm cell strainer, washed twice, resuspended in RPMI-1640 medium (Sigma-Aldrich) containing 10% fetal calf serum, 100 U/mL penicillin and 100 μg/mL streptomycin, and used for cell culture.

(247) Freshly isolated LPMCs (2×10.sup.5 cells/well) are stimulated with 1 μg/mL α-CD3/α-CD28 for 48 h in the presence of either DMSO control or appropriate concentrations of compound. After 48 h, the supernatant is removed and assayed for the presence of TNFα and IFNγ by R&D ELISA. Percentage inhibition of cytokine release by the test compounds is calculated relative to the cytokine release determined for the DMSO control (100%).

(248) (p) Inhibition of Cytokine Release from Myofibroblasts Isolated from IBD Patients

(249) Myofibroblasts from inflamed IBD mucosa are isolated as follows:

(250) The mucosa is dissected and discarded and 1 mm-sized mucosal samples are cultured at 37° C. in a humidified CO.sub.2 incubator in Dulbecco's modified Eagle's medium (DMEM, Sigma-Aldrich) supplemented with 20% FBS, 1% non-essential amino acids (Invitrogen, Paisley, UK), 100 U/mL penicillin, 100 μg/mL streptomycin, 50 μg/mL gentamycin, and 1 μg/mL amphotericin (Sigma-Aldrich). Established colonies of myofibroblasts are seeded into 25-cm.sup.2 culture flasks and cultured in DMEM supplemented with 20% FBS and antibiotics to at least passage 4 to provide a sufficient quantity for use in stimulation experiments.

(251) Subconfluent monolayers of myofibroblasts are then seeded in 12-well plates at 3×10.sup.5 cells per well are starved in serum-free medium for 24 h at 37° C., 5% CO.sub.2 before being cultured for 24 h in the presence of either DMSO control or appropriate concentrations of compound. After 24 h the supernatant is removed and assayed for the presence of IL-8 and IL-6 by R&D ELISA. Percentage inhibition of cytokine release by the test compounds is calculated relative to the cytokine release determined for the DMSO control (100%).

(252) (q) Human Neutrophil Degranulation

(253) Neutrophils are isolated from human peripheral blood as follows: Blood is collected by venepuncture and anti-coagulated by addition of 1:1 EDTA: sterile phosphate buffered saline (PBS, no Ca+/Mg+). Dextran (3% w/v) is added (1 part dextran solution to 4 parts blood) and the blood allowed to stand for approximately 20 minutes at rt. The supernatant is carefully layered on a density gradient (Lymphoprep, Axis-Shield Healthcare) and centrifuged (15 mins, 2000 rpm, no brake). The supernatant is aspirated off and the cell pellet is re-suspended in sterile saline (0.2%) for no longer than 60 seconds (to lyse contaminating red blood cells). 10 times volume of PBS is then added and the cells centrifuged (5 mins, 1200 rpm). Cells are re-suspended in HBSS+ (Hanks buffered salt solution (without phenol red) containing cytochalasin B (5 μg/mL) and 1 mM CaCl.sub.2) to achieve 5×10.sup.6 cells/mL.

(254) 5×10.sup.4 cells are added to each well of a V-bottom 96 well plate and incubated (30 mins, 37° C.) with the appropriate concentration of test compound (0.3-1000 ng/mL) or vehicle (DMSO, 0.5% final conc). Degranulation is stimulated by addition of fMLP (final conc 1 μM) which after a further incubation (30 mins, 37° C.) the cells are removed by centrifugation (5 mins, 1500 rpm) and the supernatants transferred to a flat bottom 96 well plate. An equal volume of tetramethylbenzidine (TMB) is added and after 10 mins the reaction terminated by addition of an equal volume of sulphuric acid (0.5 M) and absorbance read at 450 nm (background at 655 nm subtracted). The 50% inhibitory concentration (IC.sub.50) is determined from the resultant concentration-response curve.

(255) (r) Cell Cytotoxicity Assay

(256) 5×10.sup.4 TK6 cells (lymphoblastic T cell line) are added to the appropriate number of wells of a 96 well plate in 195 μL of media (RPMI supplemented with 10% foetal bovine serum). 5 μL of DMSO control (final concentration 0.5% v/v) or test compound (final concentration either 5 or 1 μg/mL) is added to the wells and incubated at 37° C., 5% CO.sub.2. After 24 hours, the plate is centrifuged at 1300 rpm for 3 minutes and the supernatant discarded. Cells are then resuspended in 7.5 μg/mL propidium iodide (PI) in PBS. After 15 minutes, cells are analysed by flow cytometry (BD accuri). The % viability is calculated as the % of cells that are PI negative in the test wells normalised to the DMSO control.

(257) In Vivo Screening: Pharmacodynamics and Anti-Inflammatory Activity

(258) (i) LPS-Induced Neutrophil Accumulation in Mice

(259) Non-fasted Balb/c mice are dosed by the intra tracheal route with either vehicle, or the test substance at the indicated times (within the range 2-8 hr) before stimulation of the inflammatory response by application of an LPS challenge. At T=0, mice are placed into an exposure chamber and exposed to LPS (7.0 mL, 0.5 mg/mL solution in PBS) for 30 min. After a further 8 hr the animals are anesthetized, their tracheas cannulated and BALF extracted by infusing and then withdrawing from their lungs 1.0 mL of PBS via the tracheal catheter. Total and differential white cell counts in the BALF samples are measured using a Neubaur haemocytometer. Cytospin smears of the BALF samples are prepared by centrifugation at 200 rpm for 5 min at RT and stained using a DiffQuik stain system (Dade Behring). Cells are counted using oil immersion microscopy. Data for neutrophil numbers in BAL are shown as mean±S.E.M. (standard error of the mean). The percentage inhibition of neutrophil accumulation is calculated for each treatment relative to vehicle treatment.

(260) (ii) Cigarette Smoke Model

(261) A/J mice (males, 5 weeks old) are exposed to cigarette smoke (4% cigarette smoke, diluted with air) for 30 min/day for 11 days using a Tobacco Smoke Inhalation Experiment System for small animals (Model SIS-CS; Sibata Scientific Technology, Tokyo, Japan). Test substances are administered intra-nasally (35 μL of solution in 50% DMSO/PBS) once daily for 3 days after the final cigarette smoke exposure. At 12 hr after the last dosing, each of the animals is anesthetized, the trachea cannulated and bronchoalveolar lavage fluid (BALF) is collected. The numbers of alveolar macrophages and neutrophils are determined by FACS analysis (EPICS® ALTRA II, Beckman Coulter, Inc., Fullerton, Calif., USA) using anti-mouse MOMA2 antibody (macrophage) or anti-mouse 7/4 antibody (neutrophil).

(262) (iii) DSS-Induced Colitis in Mice

(263) Non-fasted, 10-12 week old, male BDF1 mice are dosed by oral gavage twice daily with either vehicle, reference item (5-ASA) or test compound one day before (Day −1) stimulation of the inflammatory response by treatment with dextran sodium sulphate (DSS). On Day 0 of the study DSS (5% w/v) is administered in the drinking water followed by BID dosing of the vehicle (5 mL/kg), reference (100 mg/kg) or test compound (5 mg/kg) for 7 days. The drinking water with DSS is replenished every 3 days. During the study animals are weighed every day and stool observations are made and recorded as a score, based on stool consistency. At the time of sacrifice on Day +6 the large intestine is removed and the length and weight are recorded. Sections of the colon are taken for either MPO analysis to determine neutrophil infiltration or for histopathology scoring to determine disease severity.

(264) (iv) TNBS-Induced Colitis in Mice

(265) Non-fasted, 10-12 week old, male BDF1 mice are dosed by oral gavage twice daily with either vehicle (5 mL/kg), reference item (Budesonide 2.5 mg/kg) or test compound (1 or 5 mg/kg) one day before (Day −1) stimulation of the inflammatory response by treatment with 2,4,6-trinitrobenzenesulphonic acid (TNBS) (15 mg/mL in 50% ethanol/50% saline). On Day 0 of the study TNBS (200 μL) is administered intra-colonically via a plastic catheter followed by BID dosing of the vehicle, reference or test compound for 2 or 4 days. During the study animals are weighed every day and stool observations are made and recorded as a score, based on stool consistency. At the time of sacrifice on Day 2 (or Day 4) the large intestine is removed and the length and weight recorded. Sections of the colon are taken for either MPO analysis to determine neutrophil infiltration or for histopathology involving scoring to determine disease severity.

(266) (v) Adoptive Transfer in Mice

(267) On Study day 0, female Balb/C mice are terminated and spleens obtained for CD45RB.sup.high cell isolation (Using SCID IBD cell Separation protocol). Approximately 4×10.sup.5 cells/mL CD45RB.sup.high cells are then injected IP (100 μL/mouse) into female SCID animals. On study day 14, mice are weighed and randomized into treatment groups based on body weight. On Day 21 compounds are administered BID, via oral gavage, in a peanut oil vehicle at the dose levels outlined below and a dose volume of 5 mL/kg. Treatment continues until study day 42, at which point the animals are necropsied 4 hours after am administration. The colon length and weight is recorded and used as a secondary endpoint in the study as a measurement of colon oedema. The colon is then divided into six cross-sections, four of which are used for histopathology scoring (primary endpoint) and two are homogenised for cytokine analysis. Data shown is the % inhibition of the induction window between naïve animals and vehicle animals, where higher inhibition implies closer to the non-diseased, naïve, phenotype.

(268) Summary of In Vitro and In Vivo Screening Results

(269) TABLE-US-00001 TABLE 1 Results from in vitro enzyme inhibition assays Test Compound IC50 Values for Enzyme Inhibition (nM) Example No. p38 MAPKα c-Src Syk GSK3α 1 152 77 >1,000 >10,000 2 329 93 >1,000 3,237 3 347 227 >1,000 4,991 4 — — — >10,000 5 — — — 5,005 6 314 10 7 >2,000 7 380 45 673 >10,000 8 >1,000 37 269 2,153 9 358 135 520 >10,000 10 541 19 990 >10,000 11 — — — 690 12 142 10 17 1,921 13 >1,000 170 106 5,636 14 — 360 >1,000 >10,000 15 253 15 24 3,486 16 — 116 >1,000 >10,000 17 — — — >10,000 18 >1,000 41 59 >10,000 19 — — — 6,733 20 — — — 345 21 204 17 89 11,629 22 108 25 65 12,821

(270) TABLE-US-00002 TABLE 2 Results from cellular assays in d-U937 cells, PBMCs and HT29 cells (the protocols for which are described by assays (a) to (d) above). IC.sub.50 Values for Inhibition of Cytokine Release (nM) Test HT29 Compound dU937 cells PBMCs cells Example No. IL-8 TNFα IL-8 TNFα IL-2 IFNγ IL-8 1 4.6 2.5 14.2 2.8 274.1 — 15.6 2 2.6 1.8 8.4 2.7 128.3 — 12.1 3 18.0  2.0 9.4 — 76.8 — 13.4 4 — — 95.9 — — — — 5 — — 112.7 — — — — 6 1.8 1.5 4.8 0.7 15.7 — 8.7 7 8.6 1.9 12.0 3.3 47.5 — 16.6 8 — — 3.9 — — — — 9 2.6 1.2 3.4 — 41.2 — 4.8 10 2.0 1.6 3.5 — 196.1 — 5.0 11 — — 1.4 — — — — 12 — — 0.9 — 28.2 — — 13 1.4 1.1 2.3 — 3.9 — 4.3 14 — — 36.6 — — — — 15 — — 2.2 — 30.7 — 6.5 16 — — 14.8 — — — — 17 — — 8.0 — — — — 18 — — 2.7 — — — — 19 — — 76.0 — — — — 20 — — 1.8 — — — — 21 — — 2.0 — 137.4 2.0 2.0 22 3.3 0.5 1.7 — 94.8 2.3 —

(271) As illustrated in Table 3 below, the compound of Example 21 was also screened in in vivo assay (iv) above, as conducted over 2 days. Histopathology analysis revealed that the compound of Example 21 displayed significant activity in this in vivo model of colonic inflammation. In particular, these compounds, when dosed orally at 5 mg/kg, demonstrated marked improvements in ulcer grade and epithelial repair compared to the vehicle control. In addition, the compounds of Example 21 produced a marked reduction in inflammatory cell infiltrate in the reticular and laminar propria zone.

(272) TABLE-US-00003 TABLE 3 Summary of results from studies on TNBS-induced colitis in mice. TNBS Experiment LP no. Treatment group n Ulcer grade inflammation 1 Non-diseased 6 0 ± 0 0.3 ± 0.2 1 TNBS + Vehicle 12 3.7 ± 0.3 4.1 ± 0.2 1 TNBS + 5-ASA 12 3.0 ± 0.5 2.3 ± 0.3 1 TNBS + Example 21 (1 12 3.3 ± 0.5 3.8 ± 0.3 mg/kg) 1 TNBS + Example 21 (5 12 3.2 ± 0.4 2.7 ± 0.3 mg/kg)

ABBREVIATIONS

(273) 5-ASA 5-aminosalicylic acid AcOH glacial acetic acid aq aqueous ATP adenosine-5′-triphosphate BALF bronchoalveolar lavage fluid BINAP 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl br broad BSA bovine serum albumin CatCart® catalytic cartridge CDI 1,1-carbonyl-diimidazole COPD chronic obstructive pulmonary disease d doublet dba dibenzylideneacetone DBU 1,8-diazabicyclo[5.4.0]undec-7-ene DCM dichloromethane DMA dimethylacetamide DMF dimethylformamide DMSO dimethyl sulfoxide DPPA diphenylphosphoryl azide d-U937 cells PMA differentiated U-937 cells (ES.sup.+) electrospray ionization, positive mode Et ethyl Et.sub.3N triethylamine EtOAc ethyl acetate EtOH ethanol FCS foetal calf serum FRET fluorescence resonance energy transfer GSK3α glycogen synthase kinase 3α HBEC primary human bronchial epithelial cells HPLC high performance liquid chromatography hr hour(s) HRP horseradish peroxidise HRV human rhinovirus ICAM-1 inter-cellular adhesion molecule 1 JNK c-Jun N-terminal kinase LC liquid chromatography LPS lipopolysaccharide (M+H).sup.+ protonated molecular ion MAPK mitogen-activated protein kinase MAPKAP-K2 mitogen-activated protein kinase-activated protein kinase-2 Me methyl MeCN acetonitrile MeOH methanol MHz megahertz min minute(s) MMAD mass median aerodynamic diameter MOI multiplicity of infection MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MS mass spectrometry m/z: mass-to-charge ratio NBS N-bromosuccinimide NMP N-methylpyrrolidone NMR nuclear magnetic resonance (spectroscopy) PBMC peripheral blood mononuclear cell PBS phosphate buffered saline Ph phenyl PHA phytohaemagglutinin PMA phorbol myristate acetate p-TsOH 4-methylbenzenesulfonic acid (para-toluenesulfonic acid) q quartet rt room temperature RP HPLC reverse phase high performance liquid chromatography RSV respiratory syncytial virus s singlet sat saturated SCX solid supported cation exchange (resin) SDS sodium dodecyl sulphate S.sub.NAr nucleophilic aromatic substitution t triplet TBAF tetrabutylammonium fluoride TBDMS tert-butyldimethylsilyl TCID.sub.50 50% tissue culture infectious dose THF tetrahydrofuran TFA trifluoroacetic acid TNFα tumor necrosis factor alpha

(274) Prefixes n-, s-, i-, t- and tert- have their usual meanings: normal, secondary, iso, and tertiary.