WO2009056886A1

WO2009056886A1 - Pyrimidine derivatives and their use as modulators of fgfr activity

Info

Publication number: WO2009056886A1
Application number: PCT/GB2008/051018
Authority: WO
Inventors: Kevin Michael Foote; Maria-Elena Theoclitou
Original assignee: Astrazeneca Ab; Astrazeneca Uk Limited
Priority date: 2007-11-01
Filing date: 2008-10-30
Publication date: 2009-05-07

Abstract

There is provided pyrimidine compounds of formula 1: or pharmaceutical salts thereof. There is al so provided processes for their preparation, pharmaceutical compositions containing them, a process for preparing the pharmaceutical compositions, and their use in therapy, for exampl e in the treatment of proliferative disease such as cancer and particular ly in disease mediated by a FGFR inhibitory effect.

Description

PYRIMIDINE DERIVATIVES AND THEIR USE AS MODULATORS OF FGFR ACTIVITY

The present invention relates to pyrimidine derivatives, a process for their preparation, pharmaceutical compositions containing them, a process for preparing the pharmaceutical compositions, and their use in therapy.

Protein kinases are a class of proteins (enzymes) that regulate a variety of cellular functions. This is accomplished by the phosphorylation of specific amino acids on protein substrates resulting in conformational alteration of the substrate protein. The conformational change modulates the activity of the substrate or its ability to interact with other binding partners. The enzyme activity of the protein kinase refers to the rate at which the kinase adds phosphate groups to a substrate. It can be measured, for example, by determining the amount of a substrate that is converted to a product as a function of time. Phosphorylation of a substrate occurs at the active-site of a protein kinase.

Tyrosine kinases are a subset of protein kinases that catalyze the transfer of the terminal phosphate of adenosine triphosphate (ATP) to tyrosine residues on protein substrates. These kinases play an important part in the propagation of growth factor signal transduction that leads to cellular proliferation, differentiation and migration.

Fibroblast growth factor (FGF) has been recognized as an important mediator of many physiological processes, such as morphogenesis during development and angiogenesis. There are currently over 25 known members of the FGF family. The fibroblast growth factor receptor (FGFR) family consists of four members with each composed of an extracellular ligand binding domain, a single transmembrane domain and an intracellular cytoplasmic protein tyrosine kinase domain. Upon stimulation with FGF, FGFRs undergo dimerisation and transphosphorylation, which results in receptor activation. Receptor activation is sufficient for the recruitment and activation of specific downstream signalling partners that participate in the regulation of diverse process such as cell growth, cell metabolism and cell survival (Reviewed in Eswarakumar, V. P. et. al., Cytokine & Growth Factor Reviews 2005, 16, pl39-149). Consequently, FGF and FGFRs have the potential to initiate and/ or promote tumorigenesis. There is now considerable evidence directly linking FGF signalling to human cancer. The elevated expression of various FGFs has been reported in a diverse range of tumour types such as bladder, renal cell and prostate (amongst others). FGF has also been described as a powerful angiogenic factor. The expression of FGFRs in endothelial cells has also been reported. Activatiing mutations of various FGFRs have been associated with bladder cancer and multiple myeloma (amongst others) whilst receptor expression has also been documented in prostate and bladder cancer amongst others (Reviewed in Grose, R. et. al, Cytokine & Growth Factor Reviews 2005, 16, pl79-186 and Kwabi-Addo, B. et. al, Endocrine-Related Cancer 2004, 11, p709-724). For these reasons, the FGF signalling system is an attractive therapeutic target, particularly since therapies targeting FGFRs and/ or FGF signalling may affect both the tumour cells directly and tumour angiogenesis.

In accordance with the present invention, there is provided a compound of formula (I):

(I) wherein

D represents CH₂;

E represents CH₂ or O;

R^N1 represents hydrogen or a Ci-C3alkyl group optionally substituted by one or more substituents selected from Ci-C3alkoxy, cyano, hydroxyl, amino (-NH₂), mono-Ci-C₃alkylamino and di-(Ci-C₃alkyl)amino;

R¹ represents -F, -OH, -CN, a Ci-C₃alkoxy group optionally substituted by one or more R⁸ , or -CONHR⁶; R² represents hydrogen or a Ci-C3alkyl group optionally substituted by one or more substituents selected from Ci-C3alkoxy, cyano, hydroxyl, amino (-NH₂), mono-Ci-C₃alkylamino and di-(Ci-C₃alkyl)amino;

R³ represents hydrogen,

-F, or a -CONHR⁷ group; R⁴ represents hydrogen, a Ci-Cόalkyl group optionally substituted with Ci-C3alkoxy, hydroxyl, amino (-NH₂), mono-Ci-C3alkylamino and di-

(Ci-C3alkyl)amino, a C₂-C₆alkenyl group optionally substituted with Ci-C₃alkoxy, a C₂-Cόalkynyl group optionally substituted with Ci-C3alkoxy, a C₃-C₅cycloalkyl group optionally substituted with Ci-C₃alkoxy, a Ci-Cόalkoxy group optionally substituted with Ci-C3alkoxy, hydroxyl, amino (-NH₂), mono-Ci-C₃alkylamino and di-(Ci-C3alkyl)amino,

-C(O)NR⁹R¹⁰,

-NR¹¹R¹²,

-S(O)_yR¹³ where y is O, 1 or 2; R⁵ represents hydrogen or a Ci-C3alkoxy group; R⁶ represents a Ci-Cόalkyl optionally substituted by one or more R⁸; R⁷ represents hydrogen or a Ci-Cόalkyl optionally substituted by one or more R⁸; R⁹ and R¹⁰ each independently represent hydrogen, d-C₄alkyl or C₃-C₆cycloalkyl, or R⁹ and R¹⁰ together with the nitrogen atom to which they are attached form a 4- to 6-membered saturated heterocycle optionally comprising an additional heteratom selected from oxygen, sulphur or nitrogen wherein each R⁹ and R¹⁰ independently may be optionally substituted on carbon by one or more substituents R¹⁴ and wherein if said heterocycle contains an -NH- moiety that nitrogen may be optionally substited by a group selected from R¹⁵ ; R¹¹ and R¹² each independently represent hydrogen, d-C₄alkyl or C₃-C₆cycloalkyl, or R¹¹ and R¹² together with the nitrogen atom to which they are attached form a 4- to 6-membered saturated heterocycle optionally comprising an additional heteratom selected from oxygen, sulphur or nitrogen wherein each R¹¹ and R¹² independently may be optionally substituted on carbon by one or more substituents R¹⁶ and wherein if said heterocycle contains an -NH- moiety that nitrogen may be optionally substited by a group selected from R¹⁷ ; R¹³ represents Ci-Cόalkyl or C₃-C6cycloalkyl;

R⁸, R¹⁴ and R¹⁶ each independently is selected from halogen, Ci-C₃alkyl, Ci-C₃alkoxy, C₃cycloalkyl, Ci-C₃alkylthio, amino (-NH₂), mono- and di- Ci-C₃alkylamino, cyano, hydroxyl and trifluoromethyl; R¹⁵ and R¹⁷ each independently is selected from Ci-C_όalkyl, benzyl, Ci-C_όalkoxycarbonyl, Ci-C_όalkylcarbonyl, phenylcarbonyl, Ci-C_όalkylsulphonyl and phenylsulphonyl; and either when Q represents CR⁵, Q¹ represents N or CH; or when Q represents N, Q¹ represents CH, or a pharmaceutically acceptable salt thereof.

In a further aspect of the present invention, there is provided a compound of formula (I):

(I) wherein

D represents CH₂; E represents CH₂ or O;

R¹ represents -F, -OH, a Ci-C3alkoxy group optionally substituted by one or more R⁸ , or -CONHR⁶; R² represents hydrogen or a Ci-C3alkyl group optionally substituted by one or more substituents selected from Ci-C3alkoxy, cyano, hydroxyl, amino (-NH₂), mono-Ci-C₃alkylamino and di-(Ci-C₃alkyl)amino;

R³ represents hydrogen, -F, or a -CONHR⁷ group;

R⁴ represents hydrogen, a Ci-Cόalkyl group optionally substituted with Ci-C3alkoxy, hydroxyl, amino (-NH₂), mono-Ci-C3alkylamino and di- (Ci-C3alkyl)amino, a C₂-C₆alkenyl group optionally substituted with Ci-C₃alkoxy, a C₂-Cόalkynyl group optionally substituted with Ci-C3alkoxy, a C₃-C₅cycloalkyl group optionally substituted with Ci-C₃alkoxy, a Ci-Cόalkoxy group optionally substituted with Ci-C3alkoxy, hydroxyl, amino (-NH₂), mono-Ci-C₃alkylamino and di-(Ci-C3alkyl)amino, -C(O)NR⁹R¹⁰, -NR¹¹R¹²,

-S(O)_yR¹³ where y is O, 1 or 2; R⁵ represents hydrogen or a Ci-C3alkoxy group;

R⁶ represents a Ci-Cόalkyl optionally substituted by one or more R⁸;

R⁷ represents hydrogen or a Ci-Cόalkyl optionally substituted by one or more R⁸; R⁹ and R¹⁰ each independently represent hydrogen, d-C₄alkyl or C₃-C₆cycloalkyl, or R⁹ and R¹⁰ together with the nitrogen atom to which they are attached form a A- to 6-membered saturated heterocycle optionally comprising an additional heteratom selected from oxygen, sulphur or nitrogen wherein each R⁹ and R¹⁰ independently may be optionally substituted on carbon by one or more substituents R¹⁴ and wherein if said heterocycle contains an -NH- moiety that nitrogen may be optionally substited by a group selected from R¹⁵ ;

R¹¹ and R¹² each independently represent hydrogen, Ci-C₄alkyl or Cs-C_όCycloalkyl, or R¹¹ and R¹² together with the nitrogen atom to which they are attached form a A- to 6-membered saturated heterocycle optionally comprising an additional heteratom selected from oxygen, sulphur or nitrogen wherein each R¹¹ and R¹² independently may be optionally substituted on carbon by one or more substituents R¹⁶ and wherein if said heterocycle contains an -NH- moiety that nitrogen may be optionally substited by a group selected from R¹⁷ ; R¹³ represents Ci-Cβalkyl or Cs-C_όCycloalkyl;

R⁸, R¹⁴ and R¹⁶ each independently is selected from halogen, Ci-C₃alkyl, Ci-C₃alkoxy, Cscycloalkyl, Ci-C₃alkylthio, amino (-NH₂), mono- and di- Ci-C3alkylamino, cyano, hydroxyl and trifluoromethyl; R¹⁵ and R¹⁷ each independently is selected from Ci-C_όalkyl, benzyl, Ci-C_όalkoxycarbonyl, Ci-C_όalkylcarbonyl, phenylcarbonyl, Ci-C_όalkylsulphonyl and phenylsulphonyl; and either when Q represents CR⁵, Q¹ represents N or CH; or when Q represents N, Q¹ represents CH, or a pharmaceutically acceptable salt thereof. In this specification the term "alkyl" includes both straight and branched chain alkyl groups. References to individual straight chain alkyl groups such as "n-propyl" are specific for the straight chain version only and references to individual branched chain alkyl groups such as "isopropyl" or "/-propyl" are specific for the branched chain version only. Examples of "Ci.C_όalkyl" include methyl, ethyl, n-propyl, /-propyl, butyl, /-butyl, t- butyl, n-pentyl, /-pentyl, neopentyl and hexyl. Examples

include methyl, ethyl, n-propyl, /-propyl, n-butyl, /-butyl and t-butyl. A similar convention applies to other radicals, for example, examples of "Ci.Cόalkoxycarbonyl" include methoxycarbonyl, ethoxycarbonyl, n-propoxycarbonyl, z^'-propoxycarbonyl, butoxycarbonyl, i- butoxycarbonyl, t-butoxycarbonyl, n-pentoxycarbonyl, z^'-pentoxycarbonyl, neopentoxycarbonyl and hexoxycarbonyl. Examples of "Ci.Cόalkoxy" include methoxy, ethoxy, n-propoxy, z-propoxy, n-butoxy, z^'-butoxy, t-butoxy, pentoxy, z^'-pentoxy, neopentoxy, hexoxy). Examples of "Ci.Csalkoxy" include methoxy, ethoxy, n-propoxy and z^'-propoxy. Example of "Ci-Cόalkylthio" include methylthio, ethylthio, n-propylthio, i- propylthio, n-butylthio, z^'-butylthio, t-butylthio, pentylthio, z^'-pentylthio, neopentylthio and hexylthio. Examples of "Ci.Cόalkylcarbonylamino" include formamido, acetamido and propionylamino. Examples of "S(O)_mCi-C₆alkyr, "S(O)_xCi.C₆alkyl" and "S(O)_yCi.C₆alkyl" wherein m is 0, 1 or 2 include Ci-C_όalkylthio, Ci-C_όalkylsulphinyl and Ci-C_όalkylsulphonyl, for example methylthio, ethylthio, methylsulphinyl, ethylsulphinyl, mesyl and ethylsulphonyl. Examples of "Ci.Cόalkylcarbonyl" include methylcarbonyl (acetyl), ethylcarbonyl (propionyl), propylcarbonyl, z^'-propylcarbonyl, butylcarbonyl, z^'-butylcarbonyl, t-butylcarbonyl, pentylcarbonyl, z^'-pentylcarbonyl, neopentylcarbonyl and hexylcarbonyl. Examples of "^-C_όalkenyl" include vinyl, allyl, 1-propenyl, butenyl and isobutenyl. Examples of "C₂-C₆alkynyl" include acetylenyl and propargyl. Examples of "mono- and di-Ci.C_όalkylamino" include methylamino, ethylamino, n-propylamino, i- propylamino, n-butylamino, /-butylamino, t-butylamino, n-pentylamino, /-pentylamino, neopentylamino, hexylamino, dime thy lamino, diethylamino and ethylmethylamino. Where optional substituents are chosen from "one or more" groups it is to be understood that this definition includes all substituents being chosen from one of the specified groups or the substituents being chosen from two or more of the specified groups. The term "halo" refers to fiuoro, chloro, bromo and iodo. The term "amino" refers to a -NH₂ group. A "heterocyclyl" is a saturated, partially saturated or unsaturated, mono or bicyclic ring containing 4-12 atoms of which at least one atom is chosen from nitrogen, sulphur or oxygen, which may, unless otherwise specified, be carbon or nitrogen linked, wherein a -CH₂- group can optionally be replaced by a -C(O)-, and a ring sulphur atom may be optionally oxidised to form the S-oxides. Examples and suitable values of the term "heterocyclyl" are morpholino, piperidyl, pyridyl, pyranyl, pyrrolyl, pyrazolyl, isothiazolyl, indolyl, quinolyl, thienyl, 1,3-benzodioxolyl, thiadiazolyl, piperazinyl, thiazolidinyl, pyrrolidinyl, thiomorpholino, pyrrolinyl, homopiperazinyl, 3,5-dioxapiperidinyl, tetrahydropyranyl, imidazolyl, pyrimidyl, pyrazinyl, pyridazinyl, isoxazolyl, 7V-methylpyrrolyl, 4-pyridone, 1-isoquinolone, 2-pyrrolidone, 4-thiazolidone, pyridine-7V-oxide and quinoline-7V-oxide.

In one aspect of the invention a "4- to 6-membered saturated heterocyclic group", is a saturated monocyclic ring containing 4, 5 or 6 atoms of which at least one is a heteroatom selected from nitrogen, oxygen and sulphur, it may, unless otherwise specified, be carbon or nitrogen linked, a -CH₂- group can optionally be replaced by a -C(O)- and a ring sulphur atom may be optionally oxidised to form the S-oxides. Suitable "4- to 6- membered heterocyclic group" which may comprise at least one ring heteroatom selected from nitrogen, oxygen and sulphur" include tetrahydrofuran, tetrahydrofuranone, gamma- butyrolactone, dioxolane, tetrahydropyran, dioxane, thiolan, dithiolan, pyrrolidine, pyrazolidine, imidazolidine, piperidine, piperazine, morpholine, thiomorpholine, thiomorpholine S,S-dioxide, oxetane and azetidine.

In one aspect of the invention a "Cs.CόCycloalkyl" is a saturated monocyclic ring containing 3 or 6 atoms. Examples of "Cs-C_όCycloalkyl" include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

When R³ represents a 3- to 5-membered saturated heterocyclyl group, examples of the 3- to 5-membered saturated heterocyclyl group include oxirane, aziridine, azetidine and pyrrolidine. When R⁹ and R¹⁰, or R¹¹ and R¹² represent a saturated heterocycle, it should be understood that when the heterocycle comprises only one heteroatom, the heteroatom present is the nitrogen atom to which R⁹ and R¹⁰, or R¹¹ and R¹² are attached. Examples of 4- to 6-membered saturated heterocycles include pyrrolidinyl, piperidinyl and morpholinyl.

Particular values of variable groups are as follows. Such values may be used where appropriate with any of the definitions, claims or embodiments defined hereinbefore or hereinafter.

In one embodiment of the invention, E represents CH₂.

In another embodiment of the invention, R^N1 represents hydrogen or a Ci-C₃alkyl group (such as methyl, ethyl, n-propyl, or isopropyl). In a further aspect of the invention, R^N1 represents hydrogen or methyl.

In a further aspect of the invention, R^N1 represents hydrogen.

In one embodiment of the invention, R¹ represents a -F, -CN, -OH, -OMe or a -CONHMe group. In one embodiment of the invention, R¹ represents a -F, -OH, -OMe or a -CONHMe group.

In another embodiment of the invention, R² represents hydrogen or a Ci-C3alkyl group (such as methyl, ethyl, n-propyl, or isopropyl).

In a further aspect of the invention, R² represents hydrogen or methyl. In a further aspect of the invention, R² represents hydrogen. In a further embodiment of the invention, R³ represents hydrogen, -F or a -CONH₂ group.

In a further embodiment of the invention R⁴ hydrogen, a Ci-C_όalkyl group; a C₃-C₅cycloalkyl; a Ci-Cβalkoxy group.

In a further aspect of the invention, R⁴ represents hydrogen, methyl or methoxy.

In a further aspect R⁴ represents hydrogen. In one embodiment of the invention, R⁵ represents hydrogen or methoxy.

In a further aspect of the invention, there is provided a compound of formula (I) (as depicted above) wherein:

D represents CH₂;

E represents CH₂ or O; R^N1 represents hydrogen;

R¹ represents -F, -CN, -OH, -OMe or a -CONHMe group;

R² represents hydrogen;

R³ represents hydrogen, -F or a -CONH₂ group;

R⁴ represents hydrogen; and Q represents N or CR⁵ where R⁵ is hydrogen or -OMe, or a pharmaceutically acceptable salt thereof.

In a further aspect of the invention, there is provided a compound of formula (I) (as depicted above) wherein: D represents CH₂;

E represents CH₂ or O; R^N1 represents hydrogen; R¹ represents -F, -OH, -OMe or a -CONHMe group;

R represents hydrogen;

R³ represents hydrogen, -F or a -CONH₂ group;

D represents CH₂; E represents CH₂;

R^N1 represents hydrogen;

R¹ represents -F, -OH, -OMe or a -CONHMe group;

R represents hydrogen;

R³ represents hydrogen, -F or a -CONH₂ group; R⁴ represents hydrogen; and

Q represents N or CR⁵ where R⁵ is hydrogen or -OMe, or a pharmaceutically acceptable salt thereof.

E represents CH₂;

R^N1 represents hydrogen;

R¹ represents -OMe or a -CONHMe group;

R² represents hydrogen; R³ represents hydrogen, -F or a -CONH₂ group;

R⁴ represents hydrogen; and

Q¹ represents CH;

Q represents CR⁵ wherein when R¹ is -OMe, R⁵ is hydrogen or -OMe, and when R¹ is -CONHMe, R⁵ is hydrogen, or a pharmaceutically acceptable salt thereof. In a further aspect of the invention, there is provided a compound of formula (Ia) wherein:

(Ia) wherein:

D represents CH₂;

E represents CH₂ or O;

R^N1 represents hydrogen;

R¹ represents -F, -CN, -OH, -OMe or a -CONHMe group;

R² represents hydrogen or methyl;

R⁴ represents hydrogen;

Q represents CR⁵ where R⁵ is hydrogen or -OMe; and either when R ,3a^a represents hydrogen, R ,3b represents hydrogen or a -CONH₂ group; or when R^3a represents -F, R^3a represents hydrogen; or a pharmaceutically acceptable salt thereof.

In a further aspect of the invention, there is provided a compound of formula (Ia) wherein:

(Ia) wherein:

D represents CH₂;

E represents CH₂ or O;

R^N1 represents hydrogen;

R¹ represents -F, -OH, -OMe or a -CONHMe group;

R² represents hydrogen;

R⁴ represents hydrogen;

Q represents CR⁵ where R⁵ is hydrogen or -OMe; and either when R ,3a^a represents hydrogen, R ,3b represents hydrogen or a -CONH₂ group; or when R >3a^a _. represents -F, R ,3a^a represents hydrogen; or a pharmaceutically acceptable salt thereof.

(Ia) wherein:

D represents CH₂; E represents CH₂;

R^N1 represents hydrogen;

R¹ represents -F, -OH, -OMe or a -CONHMe group;

R² represents hydrogen;

R⁴ represents hydrogen; Q represents CR⁵ where R⁵ is hydrogen or -OMe; and either when R^3a represents hydrogen, R^3b represents hydrogen or a -CONH₂ group; or when R^3a represents -F, R^3a represents hydrogen; or a pharmaceutically acceptable salt thereof.

Examples of compounds of the invention include: N'-[5-[2-(3,5-dimethoxyphenyl)ethyl]-2H-pyrazol-3-yl]-N-(pyridin-2- ylmethyl)pyrimidine-2,4-diamine;

N'-[5-[2-(3-methoxyphenyl)ethyl]-2H-pyrazol-3-yl]-N-(pyridin-2-ylmethyl)pyrimidine- 2,4-diamine;

N-[l-(5-fluoropyridin-2-yl)ethyl]-N'-[5-[2-(3-methoxyphenyl)ethyl]-2H-pyrazol-3- yl]pyrimidine-2,4-diamine;

N'- [5 -[(3 -fluorophenyl)methoxy] - 1 H-pyrazol-3 -yl] -N-(pyridin-2-ylmethyl)pyrimidine-2,4- diamine;

3-[2-[5-[[2-(pyridin-2-ylmethylamino)pyrimidin-4-yl]amino]-2H-pyrazol-3- yl]ethyl]phenol; N'-[5-[(3,5-dimethoxyphenyl)methoxy]-lH-pyrazol-3-yl]-N-(pyridin-2- ylmethyl)pyrimidine-2,4-diamine;

6-[[[4-[[5-[2-(3-methoxyphenyl)ethyl]-2H-pyrazol-3-yl]amino]pyrimidin-2- yl]amino]methyl]pyridine-2-carboxamide; 6-[[[4-[[5-[2-(3,5-dimethoxyphenyl)ethyl]-2H-pyrazol-3-yl]amino]pyrimidin-2- yl]amino]methyl]pyridine-2-carboxamide;

6-[[[4-[[5-[2-(4-methoxypyridin-2-yl)ethyl]-2H-pyrazol-3-yl]amino]pyrimidin-2- yl]amino]methyl]pyridine-2-carboxamide;

6- [ [[4- [ [5 - [2-[3 -(methylcarbamoyl)phenyl]ethyl] - 1 H-pyrazol-3 -yl] amino]pyrimidin-2- yl]amino]methyl]pyridine-2-carboxamide;

6- [ [[4- [ [5 - [2-(3 -hydroxyphenyl)ethyl]-2H-pyrazol-3 -yl] amino]pyrimidin-2- yl]amino]methyl]pyridine-2-carboxamide;

(S)-3-(2-(5 -(2-(I -(5 -fluoropyridin-2-yl)ethylamino)pyrimidin-4-ylamino)-l H-pyrazol-3 - yl)ethyl)benzonitrile; (S)-N2-(l-(5-fluoropyridin-2-yl)ethyl)-N4-(3-(2-(pyridin-3-yl)ethyl)-lH-pyrazol-5- yl)pyrimidine-2,4-diamine; and

(S)-N2-(l-(5-fluoropyridin-2-yl)ethyl)-N4-(3-(3-methoxyphenethyl)-lH-pyrazol-5- yl)pyrimidine-2,4-diamine, and pharmaceutically acceptable salts of any one thereof. In another aspect of the invention, particular compounds of the invention are any one of the Examples or pharmaceutically acceptable salts of any one thereof.

In a further aspect of the invention, there is provided a compound selected from any one of the Examples.

The present invention further provides a process for the preparation of a compound of formula (I) as defined herinbefore above, or a pharmaceutically acceptable salt or solvate thereof, which comprises:

(I) i) reacting a compound of formula (II)

(H) wherein X represents a leaving group (e.g. halogen or sulfanyl such as methanesulfanyl or sulphonyloxy such as methanesulphonyloxy or toluene-4-sulphonyloxy), Z represents hydrogen or a halogen, and R¹, D, E, Q and R⁴ are as hereinbefore defined for a compound formula (I) with a compound of formula (III)

(HI) wherein R² and R³ are as defined hereinbefore for a compound of formula (I) to give, when Z is hydrogen, a compound of formula(I) or, when Z is halogen, a compound of formula (IV)

(IV) and (ii) when Z is a halogen, optionally reacting a compound of formula (IV) with a de- halogenating reagent to give a compound of formula (I); and optionally after (i) or (ii) carrying out one or more of the following:

• converting the compound obtained to a further compound of the invention • forming a pharmaceutically acceptable salt or solvate of the compound.

Step (i) may conveniently be carried out in a suitable solvent such as 2- methoxyethanol, 1-methylpyrrolidinone, butanol or dimethylacetamide at a temperature in the range from 90-200⁰C, optionally with microwave irradiation. The reaction can be carried out in the presence or absence of a suitable acid or base for example an inorganic acid such as hydrochloric acid or sulphuric acid, or an organic acid such as acetic acid or formic acid (or a suitable Lewis acid) or an inorganic base such as sodium carbonate, or an organic base such as 7V,7V-diisopropylethylamine.

Optional dehalogenation may conveniently be carried out in a suitable solvent such as ethanol in the presence of a suitable catalyst such as 5-20% palladium on carbon under an atmosphere of hydrogen.

Compounds of formula (II) may be prepared by reacting a compound of formula (V)

(V) wherein R¹, Q, D and E are as defined hereinbefore for a compound of formula (I), with a compound of formula (VI),

(VI) wherein X and Y each independently represents a leaving group (e.g. halogen or sulfanyl such as methanesulfanyl or sulphonyloxy such as methanesulphonyloxy or toluene-4-sulphonyloxy), Z represents hydrogen or a halogen, and R⁴ is as defined hereinbefore for a compound of formula (I) to give a compound of formula (II)

(H)

This reaction may conveniently be carried out in the presence of a suitable solvent such as ethanol, butanol, toluene or l-methylpyrrolid-2-one, optionally in the presence of a suitable acid or base for example an inorganic acid such as hydrochloric acid or sulphuric acid, or an organic acid such as acetic acid or formic acid (or a suitable Lewis acid) or an inorganic base such as sodium carbonate, or an organic base such as N,N- diisopropylethylamine and at a temperature in the range from 0⁰C to reflux.

Compounds of formula (V) where E represents CH₂ may be prepared by reacting a compound of formula (VII)

with a hydrazine of formula (VIII)

² NH Q wherein Q³ is hydrogen or a protecting group, such as an alkyl group or Boc group.

The reaction may be conveniently carried out in a solvent, such as ethanol, at temperature range of 60 to 80⁰C.

Alternatively, compounds of formula (V) where E represents O may be prepared by reacting a compound of formula (IX)

with a compound of formula (X)

wherein Q⁴ is hydrogen or a protecting group, such as an alkyl group or Boc group.

The reaction may be conveniently carried out in a solvent, such as dichloromethane, at temperature range of 0⁰C to room temperature. The reaction typically may be carried out in the presence of diisopropylazidocarboxylate and triphenylphosphine. In a further aspect of the present invention there is provide a process for the preparation of a compound of formula (I) as defined hereinbefore above, or a pharmaceutically acceptable salt or solvate thereof, which comprises: reacting a compound of formula (XI),

(XI) wherein Y is a leaving group such as chloro, and R², R³ and R⁴ are as defined hereinbefore for a compound of formula (I), with a compound of formula (V)

(V) wherein R¹, Q, Q¹, D and E are as defined hereinbefore for a compound of formula

(I) and optionally carrying out one or more of the following:

• converting the compound obtained to a further compound of the invention

• forming a pharmaceutically acceptable salt or solvate of the compound.

The process may conveniently be carried out in a suitable solvent such as 1- methylpyrrolidinone or dimethylacetamide in the presence of a suitable acid such as hydrogen chloride in dioxane at a temperature in the range from 90 to 12O⁰C.

Compounds of Formula (XI) may be prepared by (a) reacting a compound of formula (XII)

wherein R is as defined hereinbefore for a compound of formula (I) and X represents a leaving group (e.g. halogen or sulfanyl such as methanesulfanyl or sulphonyloxy such as methanesulphonyloxy or toluene-4-sulphonyloxy), with a compound of formula (III)

(III) wherein R² and R³ are as defined hereinbefore for a compound of formula (I) to give a compound of formula (XIII)

(XIII) and,

(b) by reacting a compound of formula (XIII) with a chlorinating agent to a compound of formula (XI)

(XI) wherein Y is a leaving group such as chloro.

Step (a) may conveniently be carried out in a suitable solvent such as diglyme in the presence of a suitable base such as 7V,7V-diisopropylethylamine at a temperature in the range from 120 to 18O⁰C.

Step (b) may conveniently be carried out in a suitable solvent such as toluene with a suitable chlorinating agent such as phosphorus oxychloride in the presence of a suitable base such as 7V,7V-diisopropylethylamine at a temperature in the range from 60 to 100⁰C.

In a still further aspect of the present invention there is provided a process for the preparation of a compound of formula (I) as hereinbefore defined but wherein R⁴ represent a Ci-Cβalkoxy group optionally substituted with Ci-C₃alkoxy, hydroxyl, amino (-NH₂), mono-Ci-C₃alkylamino and di-(Ci-C₃alkyl)amino; -NR¹¹R¹², or -S(O)_yR¹³, or a pharmaceutically acceptable salt or solvate thereof, which comprises: reacting a compound of formula (XIV)

(XIV) wherein A represents a leaving group (such as halogen or sulfanyl such as methanesulfanyl or sulphonyloxy such as methanesulphonyloxy or toluene-4-sulphonyloxy) with a compound of formula (XV)

H-R⁴ (XV) wherein R⁴ represents a Ci-Cβalkoxy group optionally substituted with Ci-C₃alkoxy, hydroxyl, amino (-NH₂), mono-Ci-C₃alkylamino and di-(Ci-C₃alkyl)amino; -NR¹¹R¹², or -S(O)_yR¹³ wherein y=0, and when R⁴ is -S(O)_yR¹³ wherein y=0, optionally reacting with an oxidising agent, and optionally carrying out one or more of the following:

The reaction may conveniently be carried out in a suitable solvent such as 1- methylpyrrolidinone, dimethylacetamide or a compound of formula (XV) used as solvent in the presence of a suitable base such as 7V,7V-diisopropylethylamine or sodium hydride at a temperature in the range from 80 to 200⁰C, optionally with microwave irradiation. The compound of formula (XIV) may be obtained by:

(1) reacting a compound of formula (XVI)

(XVI) wherein X, Y and A each independently represents a leaving group (such as halogen or sulfanyl such as methanesulfanyl or sulphonyloxy such as methanesulphonyloxy or toluene-4-sulphonyloxy), with a compound of formula (V),

(V) wherein R¹, Q, D and E are as defined hereinbefore for a compound of formula (I) to give a compound of formula (XVII)

(XVII) and,

(2) reacting a compound of formula (XVII) with a compound of formula (III)

(III) wherein R² and R³ are as defined hereinbefore for a compound of formula (I) to give a compound of formula (XIV)

(XIV) Step (1) may conveniently be carried out in a suitable solvent such as ethanol in the presence of a suitable base such as sodium carbonate or 7V,7V-diisopropylethylamine at a temperature in the range from 0 to 25⁰C.

Step (2) may conveniently be carried out in a suitable solvent such as butanol, hexanol, 1-methylpyrrolidinone or dimethylacetamide in the presence of a suitable base such as 7V,7V-diisopropylethylamine at a temperature in the range from 80 to 12O⁰C.

Compounds of formulae (III), (VI), (VII), (VIII), (IX), (X), (XII), (XV) and (XVI) are either commercially available, are known in the literature or may be prepared using known techniques. For example, the compound of formula (VII), where E represents CH₂, may be prepared by reacting a compound of formula (XVIII)

where OR >O1 is an alkyl ester (for example as outlined in Example 1) with acetonitrile.

The reaction may be conveniently carried out in a solvent, such as toluene, using sodium hydride as a base, at a temperature ranging from room temperature to 110⁰C.

Compounds of formula (I) can be converted into further compounds of formula (I) using standard procedures. Examples of the types of conversion reactions that may be used include introduction of a substituent by means of an aromatic substitution reaction, reduction of substituents, alkylation of substituents, de-alkylation of substituents and oxidation of substituents. The reagents and reaction conditions for such procedures are well known in the chemical art. Particular examples of aromatic substitution reactions include the introduction of a nitro group using concentrated nitric acid; the introduction of an acyl group using, for example, an acyl halide and Lewis acid (such as aluminium trichloride) under Friedel Crafts conditions; the introduction of an alkyl group using an alkyl halide and Lewis acid (such as aluminium trichloride) under Friedel Crafts conditions; and the introduction of a halogeno group. Particular examples of reduction reactions include the reduction of a nitro group to an amino group by catalytic hydrogenation with a nickel catalyst or by treatment with iron in the presence of hydrochloric acid with heating or the reduction of a cyano group to an amino group by treatment with lithium aluminium hydride; particular examples of de-alkylation reactions include the conversion of a methoxy group to a hydroxyl by treatment with boron tribromide; and particular examples of oxidation reactions include oxidation of alkylthio to alkylsulphinyl or alkylsulphonyl.

It will be appreciated by those skilled in the art that in the processes of the present invention certain functional groups such as hydroxyl or amino groups in the starting reagents or intermediate compounds may need to be protected by protecting groups. Thus, the preparation of the compounds of formula (I) may involve, at various stages, the addition and removal of one or more protecting groups.

The protection and deprotection of functional groups is described in 'Protective Groups in Organic Chemistry', edited by J.W.F. McOmie, Plenum Press (1973) and 'Protective Groups in Organic Synthesis', 2nd edition, T. W. Greene and P.G.M. Wuts, Wiley-Interscience (1991).

The compounds of formula (I) above may be converted to a pharmaceutically acceptable salt thereof, preferably an acid addition salt such as a hydrochloride, hydrobromide, phosphate, acetate, fumarate, maleate, tartrate, citrate, oxalate, methanesulphonate or/?-toluenesulphonate, or an alkali metal salt such as a sodium or potassium salt.

Certain compounds of formula (I) are capable of existing in stereoisomeric forms. It will be understood that the invention encompasses the use of all geometric and optical isomers (including atropisomers) of the compounds of formula (I) and mixtures thereof including racemates.

Certain compounds of formula (I) are capable of existing in tatomeric forms. For example,

N'-[5-[2-(3-methoxyphenyl)ethyl]-2H-pyrazol-3-yl]-N-(pyridin-2- ylmethyl)pyrimidine-2,4-diamine

may also exist as the corresponding tautomer

N'- [5 -[2-(3 -methoxyphenyl)ethyl]- 1 H-pyrazol-3 -yl] -N-(pyridin-2-ylmethyl)pyrimidine-

2,4-diamine

It is understood that compounds referred to by name, unless otherwise stated, include all tautomers of the compound.

The use of tautomers and mixtures thereof also form an aspect of the present invention. The compounds of formula (I) have activity as pharmaceuticals, in particular as modulators or inhibitors of FGFR activity, and may be used in the treatment of proliferative and hyperproliferative diseases/conditions, examples of which include the following cancers:

(1) carcinoma, including that of the bladder, brain, breast, colon, kidney, liver, lung, ovary, pancreas, prostate, stomach, cervix, colon, thyroid and skin;

(2) hematopoietic tumors of lymphoid lineage, including acute lymphocytic leukaemia, B-cell lymphoma and Burketts lymphoma;

(3) hematopoietic tumours of myeloid lineage, including acute and chronic myelogenous leukaemias and promyelocytic leukaemia; (4) tumours of mesenchymal origin, including fibrosarcoma and rhabdomyosarcoma; and

(5) other tumours, including melanoma, seminoma, tetratocarcinoma, neuroblastoma and glioma.

The compounds of the invention are especially useful in the treatment of tumors of the breast and prostate.

Thus, the present invention provides a compound of formula (I), or a pharmaceutically-acceptable salt thereof, as hereinbefore defined for use in therapy. In a further aspect, the present invention provides the use of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as hereinbefore defined in the manufacture of a medicament for use in therapy.

In the context of the present specification, the term "therapy" also includes "prophylaxis" unless there are specific indications to the contrary. The terms "therapeutic" and "therapeutically" should be construed accordingly.

The invention also provides a method of treating cancer which comprises administering to a patient in need thereof a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as hereinbefore defined. The invention still further provides a method of modulating FGFR activity which comprises administering to a patient in need thereof a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as hereinbefore defined.

We have found that the compounds defined in the present invention, or a pharmaceutically acceptable salt thereof, are effective anti-cancer agents which property is believed to arise from their FGFR inhibitory properties. Accordingly the compounds of the present invention are expected to be useful in the treatment of diseases or medical conditions mediated alone or in part by FGFR, i.e. the compounds may be used to produce a FGFR inhibitory effect in a warm-blooded animal in need of such treatment. Thus the compounds of the present invention provide a method for treating cancer characterised by inhibition of FGFR, i.e. the compounds may be used to produce an anticancer effect mediated alone or in part by the inhibition of FGFR.

Such a compound of the invention is expected to possess a wide range of anticancer properties as activating mutations in FGFR have been observed in many human cancers, including but not limited to, melanoma, papillary thyroid tumours, cholangiocarcinomas, colon, ovarian and lung cancers. Thus it is expected that a compound of the invention will possess anti-cancer activity against these cancers. It is in addition expected that a compound of the present invention will possess activity against a range of leukaemias, lymphoid malignancies and solid tumours such as carcinomas and sarcomas in tissues such as the liver, kidney, bladder, prostate, breast and pancreas. In particular such compounds of the invention are expected to slow advantageously the growth of primary and recurrent solid tumours of, for example, the breast and prostate. More particularly such compounds of the invention, or a pharmaceutically acceptable salt thereof, are expected to inhibit the growth of those primary and recurrent solid tumours which are associated with FGFR, especially those tumours which are significantly dependent on FGFR for their growth and spread, including for example, certain tumours of the breast and prostate.

Thus according to this aspect of the invention there is provided a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore for use as a medicament.

According to a further aspect of the invention there is provided the use of a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore in the manufacture of a medicament for use in the production of a FGFR inhibitory effect in a warm-blooded animal such as man.

According to this aspect of the invention there is provided the use of a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore in the manufacture of a medicament for use in the production of an anti-cancer effect in a warm-blooded animal such as man.

According to a further feature of the invention, there is provided the use of a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined herein before in the manufacture of a medicament for use in the treatment of melanoma, papillary thyroid tumours, cholangiocarcinomas, colon cancer, ovarian cancer, lung cancer, leukaemias, lymphoid malignancies, carcinomas and sarcomas in the liver, kidney, bladder, prostate, breast and pancreas, and primary and recurrent solid tumours of the skin, colon, thyroid, lungs and ovaries.

According to a further aspect of the invention there is provided the use of a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore in the production of a FGFR inhibitory effect in a warm-blooded animal such as man.

According to this aspect of the invention there is provided the use of a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined hereinbefore in the production of an anti-cancer effect in a warm-blooded animal such as man.

According to a further feature of the invention, there is provided the use of a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined herein before in the treatment of melanoma, papillary thyroid tumours, cholangiocarcinomas, colon cancer, ovarian cancer, lung cancer, leukaemias, lymphoid malignancies, carcinomas and sarcomas in the liver, kidney, bladder, prostate, breast and pancreas, and primary and recurrent solid tumours of the skin, colon, thyroid, lungs and ovaries.

According to a further feature of this aspect of the invention there is provided a method for producing a FGFR inhibitory effect in a warm-blooded animal, such as man, in need of such treatment which comprises administering to said animal an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as defined above.

According to a further feature of this aspect of the invention there is provided a method for producing an anti-cancer effect in a warm-blooded animal, such as man, in need of such treatment which comprises administering to said animal an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as defined above.

According to an additional feature of this aspect of the invention there is provided a method of treating melanoma, papillary thyroid tumours, cholangiocarcinomas, colon cancer, ovarian cancer, lung cancer, leukaemias, lymphoid malignancies, carcinomas and sarcomas in the liver, kidney, bladder, prostate, breast and pancreas, and primary and recurrent solid tumours of the skin, colon, thyroid, lungs and ovaries, in a warm-blooded animal, such as man, in need of such treatment which comprises administering to said animal an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined herein before.

In a further aspect of the invention there is provided a pharmaceutical composition which comprises a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined herein before in association with a pharmaceutically-acceptable diluent or carrier for use in the production of a FGFR inhibitory effect in a warm-blooded animal such as man.

In a further aspect of the invention there is provided a pharmaceutical composition which comprises a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined herein before in association with a pharmaceutically-acceptable diluent or carrier for use in the production of an anti-cancer effect in a warm-blooded animal such as man.

In a further aspect of the invention there is provided a pharmaceutical composition which comprises a compound of the formula (I), or a pharmaceutically acceptable salt thereof, as defined herein before in association with a pharmaceutically-acceptable diluent or carrier for use in the treatment of melanoma, papillary thyroid tumours, cholangiocarcinomas, colon cancer, ovarian cancer, lung cancer, leukaemias, lymphoid malignancies, carcinomas and sarcomas in the liver, kidney, bladder, prostate, breast and pancreas, and primary and recurrent solid tumours of the skin, colon, thyroid, lungs and ovaries in a warm-blooded animal such as man.

The compounds of formula (I) and pharmaceutically acceptable salts thereof may be used on their own but will generally be administered in the form of a pharmaceutical composition in which the formula (I) compound/salt/solvate (active ingredient) is in association with a pharmaceutically acceptable adjuvant, diluent or carrier. Depending on the mode of administration, the pharmaceutical composition will preferably comprise from 0.05 to 99 %w (per cent by weight), more preferably from 0.05 to 80 %w, still more preferably from 0.10 to 70 %w, and even more preferably from 0.10 to 50 %w, of active ingredient, all percentages by weight being based on total composition.

The present invention also provides a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, as hereinbefore defined, in association with a pharmaceutically acceptable adjuvant, diluent or carrier.

The invention further provides a process for the preparation of a pharmaceutical composition of the invention which comprises mixing a compound of formula (I), or a pharmaceutically acceptable salt thereof, as hereinbefore defined, with a pharmaceutically acceptable adjuvant, diluent or carrier.

The pharmaceutical compositions may be administered topically (e.g. to the skin or to the lung and/or airways) in the form, e.g., of creams, solutions, suspensions, heptafiuoroalkane aerosols and dry powder formulations; or systemically, e.g. by oral administration in the form of tablets, capsules, syrups, powders or granules; or by parenteral administration in the form of solutions or suspensions; or by subcutaneous administration; or by rectal administration in the form of suppositories; or transdermally. The compositions of the invention may be obtained by conventional procedures using conventional pharmaceutical excipients, well known in the art. Thus, compositions intended for oral use may contain, for example, one or more colouring, sweetening, flavouring and/or preservative agents. Suitable pharmaceutically acceptable excipients for a tablet formulation include, for example, inert diluents such as lactose, sodium carbonate, calcium phosphate or calcium carbonate, granulating and disintegrating agents such as corn starch or algenic acid; binding agents such as starch; lubricating agents such as magnesium stearate, stearic acid or talc; preservative agents such as ethyl or propyl p_-hydroxybenzoate, and anti-oxidants, such as ascorbic acid. Tablet formulations may be uncoated or coated either to modify their disintegration and the subsequent absorption of the active ingredient within the gastrointestinal tract, or to improve their stability and/or appearance, in either case, using conventional coating agents and procedures well known in the art.

Compositions for oral use may be in the form of hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oil such as peanut oil, liquid paraffin, or olive oil.

Aqueous suspensions generally contain the active ingredient in finely powdered form together with one or more suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents such as lecithin or condensation products of an alkylene oxide with fatty acids (for example polyoxethylene stearate), or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives (such as ethyl or propyl p_-hydroxybenzoate, anti-oxidants (such as ascorbic acid), colouring agents, flavouring agents, and/or sweetening agents (such as sucrose, saccharine or aspartame).

Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil (such as arachis oil, olive oil, sesame oil or coconut oil) or in a mineral oil (such as liquid paraffin). The oily suspensions may also contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set out above, and flavouring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water generally contain the active ingredient together with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above.

Additional excipients such as sweetening, flavouring and colouring agents, may also be present. The pharmaceutical compositions of the invention may also be in the form of oil-in- water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, or a mineral oil, such as for example liquid paraffin or a mixture of any of these.

Suitable emulsifying agents may be, for example, naturally-occurring gums such as gum acacia or gum tragacanth, naturally-occurring phosphatides such as soya bean, lecithin, an esters or partial esters derived from fatty acids and hexitol anhydrides (for example sorbitan monooleate) and condensation products of the said partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening, flavouring and preservative agents.

Syrups and elixirs may be formulated with sweetening agents such as glycerol, propylene glycol, sorbitol, aspartame or sucrose, and may also contain a demulcent, preservative, flavouring and/or colouring agent.

The pharmaceutical compositions may also be in the form of a sterile injectable aqueous or oily suspension, which may be formulated according to known procedures using one or more of the appropriate dispersing or wetting agents and suspending agents, which have been mentioned above. A sterile injectable preparation may also be a sterile injectable solution or suspension in a non- toxic parenterally-accep table diluent or solvent, for example a solution in 1,3-butanediol. Suppository formulations may be prepared by mixing the active ingredient with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Suitable excipients include, for example, cocoa butter and polyethylene glycols. Topical formulations, such as creams, ointments, gels and aqueous or oily solutions or suspensions, may generally be obtained by formulating an active ingredient with a conventional, topically acceptable, vehicle or diluent using conventional procedure well known in the art.

Compositions for administration by insufflation may be in the form of a finely divided powder containing particles of average diameter of, for example, 30μ or much less, the powder itself comprising either active ingredient alone or diluted with one or more physiologically acceptable carriers such as lactose. The powder for insufflation is then conveniently retained in a capsule containing, for example, 1 to 50mg of active ingredient for use with a turbo-inhaler device, such as is used for insufflation of the known agent sodium cromoglycate.

Compositions for administration by inhalation may be in the form of a conventional pressurised aerosol arranged to dispense the active ingredient either as an aerosol containing finely divided solid or liquid droplets. Conventional aerosol propellants such as volatile fluorinated hydrocarbons or hydrocarbons may be used and the aerosol device is conveniently arranged to dispense a metered quantity of active ingredient.

For further information on formulation the reader is referred to Chapter 25.2 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990.

The size of the dose for therapeutic purposes of a compound of the invention will naturally vary according to the nature and severity of the conditions, the age and sex of the animal or patient and the route of administration, according to well known principles of medicine.

In general, a compound of the invention will be administered so that a daily dose in the range, for example, from 0.5 mg to 75 mg active ingredient per kg body weight is received, given if required in divided doses. In general lower doses will be administered when a parenteral route is employed. Thus, for example, for intravenous administration, a dose in the range, for example, from 0.5 mg to 30 mg active ingredient per kg body weight will generally be used. Similarly, for administration by inhalation, a dose in the range, for example, from 0.5 mg to 25 mg active ingredient per kg body weight will generally be used. Oral administration is however preferred. For example, a formulation intended for oral administration to humans will generally contain, for example, from 0.5 mg to 2 g of active ingredient.

For further information on Routes of Administration and Dosage Regimes the reader is referred to Chapter 25.3 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990.

The anti cancer treatment defined hereinbefore may be applied as a sole therapy or may involve, in addition to the compound of the invention, conventional surgery or radiotherapy or chemotherapy. Such chemotherapy may include one or more of the following categories of anti-tumour agents :-

(i) other antiproliferative/antineoplastic drugs and combinations thereof, as used in medical oncology, such as alkylating agents (for example cis platin, oxaliplatin, carboplatin, cyclophosphamide, nitrogen mustard, melphalan, chlorambucil, busulphan, temozolamide and nitrosoureas); antimetabolites (for example gemcitabine and antifolates such as fluoropyrimidines like 5 fiuorouracil and tegafur, raltitrexed, methotrexate, cytosine arabinoside, and hydroxyurea); antitumour antibiotics (for example anthracyclines like adriamycin, bleomycin, doxorubicin, daunomycin, epirubicin, idarubicin, mitomycin- C, dactinomycin and mithramycin); antimitotic agents (for example vinca alkaloids like vincristine, vinblastine, vindesine and vinorelbine and taxoids like taxol and taxotere and polokinase inhibitors); and topoisomerase inhibitors (for example epipodophyllotoxins like etoposide and teniposide, amsacrine, topotecan and camptothecin);

(ii) cytostatic agents such as antioestrogens (for example tamoxifen, fulvestrant, toremifene, raloxifene, droloxifene and iodoxyfene), antiandrogens (for example bicalutamide, fiutamide, nilutamide and cyproterone acetate), LHRH antagonists or LHRH agonists (for example goserelin, leuprorelin and buserelin), progestogens (for example megestrol acetate), aromatase inhibitors (for example as anastrozole, letrozole, vorazole and exemestane) and inhibitors of 5* -reductase such as finasteride;

(iii) anti-invasion agents (for example c-Src kinase family inhibitors like 4-(6- chloro-2,3-methylenedioxyanilino)-7-[2-(4-methylpiperazin-l-yl)ethoxy]-5- tetrahydropyran-4-yloxyquinazoline (AZD0530; International Patent Application WO 01/94341) and N-(2-chloro-6-methylphenyl)-2- {6-[4-(2-hydroxyethyl)piperazin- 1 -yl]-2- methylpyrimidin-4-ylamino}thiazole-5-carboxamide (dasatinib, BMS-354825; J. Med. Chem., 2004, 47, 6658-6661), and metalloproteinase inhibitors like marimastat, inhibitors of urokinase plasminogen activator receptor function or antibodies to Heparanase);

(iv) inhibitors of growth factor function: for example such inhibitors include growth factor antibodies and growth factor receptor antibodies (for example the anti erbB2 antibody trastuzumab [Herceptin™], the anti-EGFR antibody panitumumab, the anti erbBl antibody cetuximab [Erbitux, C225] and any growth factor or growth factor receptor antibodies disclosed by Stern et al. Critical reviews in oncology/haematology, 2005, Vol. 54, pp 11-29); such inhibitors also include tyrosine kinase inhibitors, for example inhibitors of the epidermal growth factor family (for example EGFR family tyrosine kinase inhibitors such as N-(3-chloro-4-fluorophenyl)-7-methoxy-6-(3-morpholinopropoxy)quinazolin-4- amine (gefitinib, ZD 1839), N-(3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4- amine (erlotinib, OSI 774) and 6-acrylamido-N-(3-chloro-4-fluorophenyl)-7-(3- morpholinopropoxy)-quinazolin-4-amine (CI 1033), erbB2 tyrosine kinase inhibitors such as lapatinib, inhibitors of the hepatocyte growth factor family, inhibitors of the platelet- derived growth factor family such as imatinib, inhibitors of serine/threonine kinases (for example Ras/Raf signalling inhibitors such as farnesyl transferase inhibitors, for example sorafenib (BAY 43-9006)), inhibitors of cell signalling through MEK and/or AKT kinases, inhibitors of the hepatocyte growth factor family, c-kit inhibitors, abl kinase inhibitors, IGF receptor (insulin-like growth factor) kinase inhibitors; aurora kinase inhibitors (for example AZDl 152, PH739358, VX-680, MLN8054, R763, MP235, MP529, VX-528 AND AX39459) and cyclin dependent kinase inhibitors such as CDK2 and/or CDK4 inhibitors;

(v) antiangiogenic agents such as those which inhibit the effects of vascular endothelial growth factor, [for example the anti vascular endothelial cell growth factor antibody bevacizumab (Avastin™) and VEGF receptor tyrosine kinase inhibitors such as 4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(l-methylpiperidin-4-ylmethoxy)quinazoline (ZD6474; Example 2 within WO 01/32651), 4-(4-fluoro-2-methylindol-5-yloxy)-6- methoxy-7-(3-pyrrolidin-l-ylpropoxy)quinazoline (AZD2171; Example 240 within WO 00/47212), vatalanib (PTK787; WO 98/35985) and SUl 1248 (sunitinib; WO 01/60814), compounds such as those disclosed in International Patent Applications WO97/22596, WO 97/30035, WO 97/32856 and WO 98/13354 and compounds that work by other mechanisms (for example linomide, inhibitors of integrin avb3 function and angiostatin)];

(vi) vascular damaging agents such as Combretastatin A4 and compounds disclosed in International Patent Applications WO 99/02166, WO 00/40529, WO 00/41669,

WO 01/92224, WO 02/04434 and WO 02/08213;

(vii) antisense therapies, for example those which are directed to the targets listed above, such as ISIS 2503, an anti-ras antisense; (viii) gene therapy approaches, including for example approaches to replace aberrant genes such as aberrant p53 or aberrant BRCAl or BRCA2, GDEPT (gene directed enzyme pro drug therapy) approaches such as those using cytosine deaminase, thymidine kinase or a bacterial nitroreductase enzyme and approaches to increase patient tolerance to chemotherapy or radiotherapy such as multi drug resistance gene therapy; and (ix) immunotherapy approaches, including for example ex vivo and in vivo approaches to increase the immunogenicity of patient tumour cells, such as transfection with cytokines such as interleukin 2, interleukin 4 or granulocyte macrophage colony stimulating factor, approaches to decrease T cell anergy, approaches using transfected immune cells such as cytokine transfected dendritic cells, approaches using cytokine transfected tumour cell lines and approaches using anti idiotypic antibodies.

Such conjoint treatment may be achieved by way of the simultaneous, sequential or separate dosing of the individual components of the treatment. Such combination products employ the compounds of this invention within the dosage range described hereinbefore and the other pharmaceutically-active agent within its approved dosage range. In addition to their use in therapeutic medicine, the compounds of formula (I) and their pharmaceutically acceptable salts are also useful as pharmacological tools in the development and standardisation of in vitro and in vivo test systems for the evaluation of the effects of inhibitors of B-Raf in laboratory animals such as cats, dogs, rabbits, monkeys, rats and mice, as part of the search for new therapeutic agents. In the above other pharmaceutical composition, process, method, use and medicament manufacture features, the alternative and preferred embodiments of the compounds of the invention described herein also apply. Examples

The invention will now be further described with reference to the following illustrative examples in which, unless stated otherwise:

(i) temperatures are given in degrees Celsius (⁰C); operations were carried out at room or ambient temperature, that is, at a temperature in the range of 18-25°C;

(ii) organic solutions were dried over anhydrous magnesium sulphate; evaporation of solvent was carried out using a rotary evaporator under reduced pressure (600-4000

Pascals; 4.5-30mmHg) with a bath temperature of up to 60⁰C;

(iii) chromatography means flash chromatography on silica gel; thin layer chromatography (TLC) was carried out on silica gel plates;

(iv) in general, the course of reactions was followed by TLC and reaction times are given for illustration only;

(v) final products had satisfactory proton nuclear magnetic resonance (NMR) spectra and/or mass spectral data; (vi) yields are given for illustration only and are not necessarily those which can be obtained by diligent process development; preparations were repeated if more material was required;

(vii) when given, NMR data is in the form of delta values for major diagnostic protons, given in parts per million (ppm) relative to tetramethylsilane (TMS) as an internal standard, determined at 300 MHz, in DMSOd₆ unless otherwise indicated;

(viii) chemical symbols have their usual meanings; SI units and symbols are used;

(ix) solvent ratios are given in volume:volume (v/v) terms; and

(x) mass spectra (MS) data was generated on an LC/MS system where the HPLC component comprised generally either a Agilent 1100 or Waters Alliance HT (2790 & 2795) equipment and was run on a Phemonenex Gemini C18 5μm, 50 x 2 mm column (or similar) eluting with either acidic eluent (for example, using a gradient between 0 - 95% water / acetonitrile with 5% of a 1% formic acid in 50:50 water: acetonitrile (v/v) mixture; or using an equivalent solvent system with methanol instead of acetonitrile), or basic eluent

(for example, using a gradient between 0 - 95% water / acetonitrile with 5% of a 0.1% 880 Ammonia in acetonitrile mixture); and the MS component comprised generally a Waters

ZQ spectrometer. Chromatograms for Electrospray (ESI) positive and negative Base Peak

Intensity, and UV Total Absorption Chromatogram from 220-3 OOnm, are generated and values for m/z are given; generally, only ions which indicate the parent mass are reported and unless otherwise stated the value quoted is the (M+H)+ for positive ion mode and (M- H)^" for negative ion mode;

(xi) Preparative HPLC was performed on Cl 8 reversed-phase silica, for example on a Waters 'Xterra' preparative reversed-phase column (5 microns silica, 19 mm diameter, 100 mm length) using decreasingly polar mixtures as eluent, for example decreasingly polar mixtures of water (containing 1% acetic acid or 1% aqueous ammonium hydroxide (d=0.88) and acetonitrile;

(xii) the following abbreviations have been used: THF tetrahydrofuran;

DMF 7V,7V-dimethylformamide;

EtOAc ethyl acetate;

DIPEA Λ/,Λ/-diisopropylethylamine/N-ethyl-N-propan-2-yl- propan-2-amine DCM dichloromethane; and

DMSO dimethylsulphoxide.

PBS phosphate buffered saline

HEPES 7V-[2-Hydroxyethyl]piperazine-Λ/'-[2-ethanesulfonic acid]

DTT dithiothreitol ATP Adenosine Triphosphate

BSA bovine serum albumin

DMEM Dulbecco's modified Eagle's Medium

(xii) compounds are named using C-lab naming software: Openeye Lexichem version 1.4 or 1.6; (xiϋ) unless otherwise specified, starting materials are commercially available.

Table 1

Example 1

N'-[5-[2-(3,5-dimethoxyphenyl)ethyl]-2H-pyrazol-3-yl]-N-(pyridin-2- ylmethyl)pyrimidine-2,4-diamine

5-[2-(3,5-Dimethoxyphenyl)ethyl]-2H-pyrazol-3-amine (290mg, l.lδmmol) and 4-chloro- N-(pyridin-2-ylmethyl)pyrimidin-2-amine (200mg, 0.92mmol) in ethanol (5mL; containing a few drops of 4M HCl in dioxane) were refluxed for 22h and then allowed to cool. The precipitated solid was filtered, washed with ethanol and then dried to obtained the clean product as a HCl salt. The filtrate was evaporated under reduced pressure and purified by preparative HPLC using decreasingly polar mixtures of water containing 1% ammonium hydroxide and MeCN as eluents. Fractions containing the desired compound were evaporated to dryness to afford N'-[5-[2-(3,5-dimethoxyphenyl)ethyl]-2H-pyrazol-3- yl]-N-(pyridin-2-ylmethyl)pyrimidine-2,4-diamine (53mg, 10.4%) as a solid.

¹H NMR (300.132 MHz, DMSO) δ 2.79 (4H, s),3.70 (6H, s),4.61 (2H, s),6.13 (IH, s),6.24 (IH, s),6.31 (IH, t),6.37 (IH, d),7.19 - 7.25 (IH, m),7.33 (IH, d),7.72 (IH, td),7.81 (IH, d),8.47 - 8.51 (IH, m). MS: m/z 394 (MH+). (FGFR Kinase assay - Caliper IC₅₀ 0.405 μM)

4-Chloro-N-(pyridin-2-ylmethyl)pyrimidin-2-amine, used as starting material, can be prepared as follows:-

2-(Pyridin-2-ylmethylamino)pyrimidin-4-ol (1.005g, 4.95mmol) and phosphorus oxychloride (1.615mL, 17.33mmol) were heated in a round-bottomed flask to 85⁰C. The reaction mixture was cooled, and evaporated to dryness. The residue was treated with saturated sodium bicarbonate (10OmL) (taking the pH to above 7) and extracted with DCM (2 x 10OmL). The solution was evaporated and purified by silica column chromatography, eluting with MeOH (3%) DCM. The desired product was isolated as a cream solid (0.51Og, 47%).

5-[2-(3,5-Dimethoxyphenyl)ethyl]-2H-pyrazol-3-amine, used as starting material was prepared as follows:-

Acetonitrile (2.29mL, 43.61mmol, 1.2eq) was added to a slurry of sodium hydride (1.75g dispersion in mineral oil, 43.61mmol, 1.2eq) in anhydrous toluene (7OmL) and the mixture stirred at room temperature for 30 mins. Ethyl 3-(3,5-dimethoxyphenyl)propanoate (8.66g, 36.34mmol, leq) in toluene (6OmL) was added and the reaction was refiuxed for 18h. After cooling, the reaction mixture was quenched with water and the solvent was evaporated under reduced pressure. The residue was dissolved in 2M HCl (5OmL). The acidic solution was extracted with ethyl acetate. The organic extracts were combined and washed with water, brine and dried over magnesium sulphate. After filtering, the solvent was evaporated under reduced pressure to yield the crude product as a yellow oil. The oil was purified by silica column chromatography (eluting with DCM) and the desired fractions were combined and evaporated to yield a cream solid (3.76g, 44% yield). To the cream solid (3.72g, 15.96mmol, leq) in ethanol (55mL) was added hydrazine hydrate (852μl, 17.56mmol, l.leq). The reaction was refiuxed for 24h before allowing to cool. After evaporation under reduced pressure, the residue was extracted into DCM. The organic layers were washed with water, brine, dried with magnesium sulphate, filtered and evaporated under reduced pressure to afford 5-[2-(3,5-dimethoxyphenyl)ethyl]-2H- pyrazol-3 -amine as a pale yellow solid (3.76g. 42% over 2 steps).

¹H NMR (300.132 MHz, DMSO) δ 2.64 - 2.82 (4H, m), 3.71 (6H, s), 4.07 - 4.72 (2H, m), 5.20 (IH, s), 6.31 (IH, t), 6.38 (2H, d). MS: m/z 248 (MH+)

Example 2 N'- [5- [2-(3-methoxyphenyl)ethyl] -2H-pyrazol-3-yl] -N-(pyridin-2- ylmethyl)pyrimidine-2,4-diamine

2-chloro-N-[5-[2-(3-methoxyphenyl)ethyl]-2H-pyrazol-3-yl]pyrimidin-4-amine (lOOmg, 0.30mmol, leq), pyridin-2-ylmethanamine (66mg, 0.61, 2eq) and N-ethyl-N-propan-2-yl- propan-2-amine (159μl, 0.91mmol, 3eq) were combined in 2-methoxyethanol (3mL). The reaction was heated in the microwave at 200⁰C for 2h. The solvent was evaporated under reduced pressure. The crude product was purified by preparative HPLC using decreasingly polar mixtures of water (containing 1% ammonium hydroxide) and MeCN as eluents. Fractions containing the desired compound were combined and evaporated to low volume. The precipitated solid was filtered, washed with water and dried to afford N'-[5-[2-(3- methoxyphenyl)ethyl]-2H-pyrazol-3-yl]-N-(pyridin-2-ylmethyl)pyrimidine-2,4-diamine (56.3mg, 47%) as a solid. ¹H NMR (300.132 MHz, DMSO) δ 2.72 - 2.93 (4H, m), 3.73 (3H, s), 4.58 (2H, d), 6.14 - 6.32 (2H, m), 6.72 - 6.84 (3H, m), 7.12 - 7.27 (3H, m), 7.30 (IH, d), 7.71 (IH, t), 7.80 (IH, d), 8.48 (IH, d), 9.29 (IH, s), 11.85 (IH, s). MS: m/z 402 (MH+) (FGFR Kinase assay - Caliper IC₅₀0.14μM)

2-Chloro-N-[5-[2-(3-methoxyphenyl)ethyl]-2H-pyrazol-3-yl]pyrimidin-4-amine, used as starting material was prepared as follows:-

5-[2-(3-Methoxyphenyl)ethyl]-2H-pyrazol-3-amine (2.08g, 9.55mmol) and 2,4- dichloropyrimidine (l.lg, 7.98mmol, leq) were combined in ethanol (4OmL) containing N- ethyl-N-propan-2-yl-propan-2-amine (2.78mL, 15.96mmol, 2eq). The reaction was heated at 50⁰C for 7Oh. After cooling the solvent was evaporated under reduced pressure and the residue extracted into ethyl acetate (10OmL) and washed with water (2 x 5OmL) then saturated brine (5OmL). The organic layer was dried with magnesium sulphate, filtered and evaporated. Trituration with DCM produced a white precipitate which was filtered, washed with 50% ether/hexane and dried to afford 2-chloro-N-[5-[2-(3-methoxyphenyl)ethyl]-2H- pyrazol-3-yl]pyrimidin-4-amine (1.29g, 49%) as a white solid. The filtrate was evaporated and purified by silica column chromatography, eluting with 50-100% ethyl acetate in hexane. The product was obtained as a yellow solid (0.469g, 18%).

¹H NMR (300.132 MHz, DMSO) δ 2.89 (4H, s),3.73 (3H, s),6.11 (IH, s),6.73 - 6.84 (3H, m),7.20 (2H, t),8.16 (IH, d),10.27 (IH, s),12.20 (IH, s). MS: m/z 330 (MH+).

5-[2-(3-Methoxyphenyl)ethyl]-2H-pyrazol-3-amine, used as starting material was prepared as follows :-

LDA (34 mL, 67mmol, 1.3eq 2M solution in THF) was added to anhydrous THF (300 mL) and cooled to -78⁰C under a nitrogen atmosphere. To this was slowly added anhydrous acetonitrile (2.8 g, 67mmol, 1.3eq). The reaction mixture was stirred for 10 mins before the rapid addition of methyl 3-(3-methoxyphenyl)propanoate (10 g, 51.55mmol, leq) and stirred for a further 30 mins before being allowed to warm up to room temperature. The reaction was quenched with 1.0 N HCl (100 mL), extracted with diethyl ether (2 x 200 mL), dried and solvent removed in vacuo to yield a yellow gum. The crude material and hydrazine (7.5mL, 154mmol, 3eq) were dissolved in ethanol (300 mL) and heated at reflux for 3 h. The reaction was cooled and the solvent removed in vacuo to yield a viscous yellow gum. To this was added water and the product was extracted with diethyl ether (3 x 200 mL), dried and the organic solvent removed in vacuo to yield light yellow gum. Purified by silica column chromatography, eluting with 60-100% diethylether in iso- hexane then 10% methanol in ethyl acetate. Evaporated to give title compound as a pale yellow oil, which solidified on standing (6.79g, 61%).

IH NMR (CDCB 400.13MHz) δ 2.87 (4H, m), 3.56 (2H, bs), 3.78 (3H, s), 5.45 (IH, s), 6.71 (IH, m), 6.75 (2H, m), 7.20 (IH, t). MS m/z 218 (MH+).

Methyl 3-(3-methoxyphenyl)propanoate, used as starting material was prepared as follows :-

3-(3-Methoxyphenyl)propanoic acid (50 g, 0.277 mmol) was dissolved in DMF (300 mL) and to this was added potassium carbonate (58 g, 0.42 mol) and methyl iodide (20 mL, 0.30 mol). The reaction was heated at 5O⁰C for 2 h. The reaction was evaporated to a smaller volume and diethyl ether (200 mL) was added. The solution was filtered and the filtrate was evaporated to yield brown oil. This was purified via vacuum distillation at 102⁰C @ 0.33 mbar to yield a clear liquid (53.5g, 99.5%).

¹H NMR (400.132 MHz, CDC13) δ 7.20 (dd, IH), 6.79 - 6.74 (m, 3H), 3.79 (s, 3H), 3.67 (s, 3H), 2.93 (t, 2H), 2.63 (t, 2H)

Example 3

N- [ l-(5-fluoropyridin-2-yl)ethyl] -N^f- [5- [2-(3-methoxyphenyl)ethyl] -2H-pyrazol-3- yl] pyrimidine-2,4-diamine

2-chloro-N-[5-[2-(3-methoxyphenyl)ethyl]-2H-pyrazol-3-yl]pyrimidin-4-amine (lOOmg, 0.30mmol, leq) , l-(5-fluoropyridin-2-yl)ethanamine hydrochloride (81mg, 0.45mmol) and N-ethyl-N-propan-2-yl-propan-2-amine (159μl, 0.91mmol, 3eq) were combined in 2- methoxyethanol (3mL). The reaction was heated in the microwave at 200⁰C for 2h. The solvent was evaporated under reduced pressure. The crude product was purified by preparative HPLC using decreasingly polar mixtures of water containing 1% ammonium hydroxide and MeCN as eluents. Fractions containing the desired compound were combined and evaporated to afford N-[l-(5-fluoropyridin-2-yl)ethyl]-N'-[5-[2-(3- methoxyphenyl)ethyl]-2H-pyrazol-3-yl]pyrimidine-2,4-diamine (50.9mg, 39.2%) as a solid.

¹H NMR (300.132 MHz, DMSO) δ 1.46 (3H, d),2.78 - 2.95 (4H, m),3.73 (3H, s),5.13 (IH, m),6.10 - 6.28 (2H, m),6.73 - 6.85 (3H, m),7.01 (IH, s),7.20 (IH, t),7.45 (IH, dd),7.64 (IH, td),7.78 (IH, d),8.46 (IH, s),9.26 (IH, s),11.86 (IH, s). MS: m/z 434 (MH+). (FGFR Kinase assay - Caliper IC500.17 μM)

2-chloro-N-[5-[2-(3-methoxyphenyl)ethyl]-2H-pyrazol-3-yl]pyrimidin-4-amine, used as starting material was prepared as in Example 2.

l-(5-fluoropyridin-2-yl)ethanamine hydrochloride used as starting material can be prepared by the method described in the literature (Wang, Bin; Wang, Tao. 3-(Pyrazol-3- ylamino)pyrimidines as Trk A kinase inhibitors and their preparation, pharmaceutical compositions and use in the treatment of cancers.WO2006123113).

Example 4

N^f- [5- [(3-fluorophenyl)methoxy] - lH-pyrazol-3-yl] -N-(pyridin-2-ylmethyl)pyrimidine-

2,4-diamine Prepared using an analogous method to example 12, but starting with 5-[(3- fiuorophenyl)methoxy]-lH-pyrazol-3 -amine (97mg, 0.36mmol) to give the title compound (61mg, 40% yield).

IH NMR (300.132 MHz, DMSO) δ 4.74 (d, 2H), 5.14 (s, 2H), 5.82 (s, IH), 6.36 (d, IH), 7.14 - 7.20 (m, IH), 7.25 - 7.29 (m, 2H), 7.33 - 7.36 (m, 2H), 7.43 - 7.48 (m, 2H), 7.83 (t, IH), 7.92 (d, IH), 8.54 (s, IH), 9.06 (d, IH), 11.32 (s, IH) MS: m/z 392 (MH+) (FGFR Kinase assay - Caliper IC₅₀0.515μM)

5-[(3-fluorophenyl)methoxy]-lH-pyrazol-3-amine, used as starting material, was prepared as follows: -

3-Amino-5-hydroxypyrazole (2.16g, 21.82mmol, leq) and triphenylphosphine (6.88g, 26.2mmol, 1.2eq) were stirred in DCM (22mL) for 30 mins. After this time, diisopropyl azodicarboxylate (5.16mL, 26.2mmol, 1.2eq) was slowly added, keeping the temp below 20 ⁰C with a water bath, and the resulting mixture stirred for a further 45 mins. A solution of (3-fluorophenyl)methanol (3.3g, 26.2mmol) in DCM (1OmL) was added slowly and the reaction left to stir at room temperature for 24 h. After this time the solid was filtered off 5 and the solution extracted with 2M HCl solution (3x30mL). The aqueous layer was washed with diethyl ether (2x3 OmL) and then basified to pH 9 using ammonium hydroxide (with cooling). The solution was extracted with DCM (3x30mL) and the organic fractions combined, dried over magnesium sulphate and concentrated to give 5 -[(3- fiuorophenyl)methoxy]-lH-pyrazol-3 -amine as a white solid (428mg, 10%). o IH NMR (300.132 MHz, DMSO) δ 4.76 (s, IH), 4.93 (s, 2H), 5.06 (s, 2H), 7.09 - 7.15 (m, IH), 7.18 - 7.24 (m, 2H), 7.37 - 7.44 (m, IH), 10.41 (s, IH). MS: m/z 208 (MH+)

Example 5

3- [2- [5- [ [2-(pyridin-2-ylmethylamino)pyrimidin-4-yl] amino] -2H-pyrazol-3-5 yl] ethyl] phenol

To a stirred suspension of N'-[5-[2-(3-methoxyphenyl)ethyl]-lH-pyrazol-3-yl]-N-(pyridin- 2-ylmethyl)pyrimidine-2,4-diamine.2HCl (lOOmg, 021 lmmol) in DCM (1OmL), stirred at 0 ⁰C under nitrogen, was slowly added BBr₃ (l.lmL of a IM solution in DCM, 1.055mmol). The reaction was allowed to warm to room temp over 1.5h and stirring0 continued for a further 18h. The reaction mixture was then cooled in ice and methanol was slowly added (3mL) to yield a colourless solution. The solvent was removed by evaporation and the residue was dissolved in water and basified with satd aq NaHCO₃. The precipitated solid was collected by filtration, washed with water and dried under vacuum to afford the title compound as an off-white solid (76mg, 93%). 5 ¹H NMR (399.902 MHz, DMSO) δ 2.72 - 2.83 (m, 4H), 4.61 (d, 2H), 5.96 - 6.43 (bm, 2H), 6.57 - 6.62 (m, IH), 6.62 - 6.67 (m, 2H), 7.08 (t, IH), 7.22 - 7.27 (m, IH), 7.34 (d, IH), 7.54 (bs, IH), 7.71 - 7.78 (m, IH), 7.83 (d, IH), 8.51 (d, IH), 9.25 (s, IH), 9.75 (bs, IH), 11.97 (bs, IH). MS: m/z 388 (MH+) (FGFR Kinase assay - Caliper IC₅₀0.02 lμM) o

N'- [5 -[2-(3 -methoxyphenyl)ethyl]- 1 H-pyrazol-3 -yl] -N-(pyridin-2-ylmethyl)pyrimidine- 2,4-diamine.2HCl, used as starting material, was prepared as follows: - 5-[2-(3-Methoxyphenyl)ethyl]-lH-pyrazol-3-amine (130mg, 0.598mmol) was heated with 4-chloro-Λ/-(pyridin-2-ylmethyl)pyrimidin-2-amine (152mg, 0.688mmol) in ethanol (5mL) at 80⁰C for 2 days. The mixture was allowed to cool to room temperature and the precipitated solid was collected by filtration and washed with a small amount of ethanol and ether, then dried under vacuum to afford the product, N'-[5-[2-(3- methoxyphenyl)ethyl]-lH-pyrazol-3-yl]-N-(pyridin-2-ylmethyl)pyrimidine-2,4- diamine.2HCl, as an off-white solid (206mg, 73%).

¹H NMR (399.902 MHz, DMSO) δ 2.77 - 2.91 (m, 4H), 3.73 (s, 3H), 4.74 (d, 2H), 6.19 (bs, IH), 6.34 (bs, IH), 6.74 - 6.82 (m, 3H), 7.20 (t, IH), 7.32 (t, IH), 7.41 (d, IH), 7.80 (t, IH), 7.88 (bs, IH), 8.55 (d, IH), 8.76 (bs, IH), 11.14 (bs, IH), 12.36 (bs, IH). MS: m/z 402 (MH+)

4-Chloro-N-(pyridin-2-ylmethyl)pyrimidin-2-amine, used as starting material, can be prepared as in Example 1.

Example 6

N^f-[5-[(3,5-dimethoxyphenyl)methoxy]-lH-pyrazol-3-yl]-N-(pyridin-2- ylmethyl)pyrimidine-2,4-diamine

5-[(3,5-Dimethoxyphenyl)methoxy]-2H-pyrazol-3-amine.HCl (55mg, 0.192mmol) was heated with 4-chloro-Λ/-(pyridin-2-ylmethyl)pyrimidin-2-amine (49mg, 0.221mmol) in ethanol at 80⁰C for 18h. The solution was allowed to cool to room temperature and evaporated to dryness. The residue was taken up in ethyl acetate (15mL) and aqueous NaHCO₃ (15mL). The layers were separated and the aqueous phase was re-extracted with ethyl acetate (2 x 1OmL). The combined extracts were dried over MgSO₄, filtered and evaporated. The reaction was repeated as above and the combined product from the two reactions was purified on a silica isolute column, eluting with 0-3.5% methanol in DCM, to afford the title compound as a white solid (46mg, 55%).

¹H NMR (399.902 MHz, DMSO) δ 3.68 (s, 6H), 4.51 (s, 2H), 5.00 (s, 2H), 5.20 (s, IH), 5.90 (d, IH), 6.37 (t, IH), 6.53 (d, 2H), 7.16 - 7.21 (m, IH), 7.29 (d, IH), 7.65 - 7.73 (m, 2H), 7.80 (d, IH), 8.44 (d, IH), 9.89 (s, IH), 11.93 (s, IH) MS: m/z 434 (MH+) (FGFR Kinase assay - Caliper IC₅₀0.18μM) 4-Chloro-N-(pyridin-2-ylmethyl)pyrimidin-2-amine used as starting material was prepared as in Example 1.

5-[(3,5-Dimethoxyphenyl)methoxy]-2H-pyrazol-3-amine.HCl, used as starting material was synthesized as follows : -

3-Amino-5-hydroxypyrazole (8g, 80.74mmol) and triphenylphosphine (25.45g, 96.88mmol) were stirred in DCM (HOmL) under nitrogen and the mixture was cooled in an ice-bath. Diisopropylazodicarboxylate (19.08mL, 96.88mmol) was added dropwise (temperature <10°C) and the reaction mixture was stirred in the ice-bath for Ih. 3,5- Dimethoxybenzyl alcohol (16.3Og, 96.88mol) in DCM (35mL) was added dropwise, the reaction mixture was allowed to warm to room temperature and stirred under nitrogen for 4 days. The mixture was filtered, washed with DCM and the filtrate was extracted with IM HCl (aq) (3 x 5OmL). The combined aqueous extracts were washed with DCM (5OmL), resulting in precipitation of the product. The product was collected by filtration, washed with water, DCM and dried under vacuum to afford the title compound as a white solid (358mg, 1.8% yield). A further crop of product was obtained following precipitation from the initial DCM layer on allowing to stand at room temperature. The solid product was collected by filtration, washed with DCM and dried under vacuum to give an off-white solid (1.127g, 5.6% yield). 1H NMR (300.132 MHz, DMSO) δ 3.75 (s, 6H), 5.18 (s, 2H), 5.26 (s, IH), 6.50 (t, IH), 6.60 (d, 2H). MS:m/z 250 (MH+)

Example 7

6- [ [ [4- [ [5- [2-(3-methoxyphenyl)ethyl] -2H-pyrazol-3-yl] amino] pyrimidin-2- yl] amino] methyl] pyridine-2-carboxamide

A mixture of 2-chloro-N-[5-[2-(3-methoxyphenyl)ethyl]-2H-pyrazol-3-yl]pyrimidin-4- amine (66mg, 0.2mmol, l.Oeq), 6-(aminomethyl)pyridine-2-carboxamide (71mg, 0.3mmol, 1.5eq), N-ethyl-N-propan-2-yl-propan-2-amine (122μL, 0.7mmol, 3.5eq) in 2- methoxyethanol (2mL) was heated in a microwave reactor at 180⁰C for 60 mins. The resulting solution was allowed to cool to room temperature and then evaporated to dryness. This material was purified by reverse-phase preparative HPLC (basic) using a 25-45% gradient of acetonitrile in water (containing 1% ammonium hydroxide) solution. The clean fractions were taken and evaporated to afford the title compound as a solid, (46mg, 52% yield).

¹H NMR (500.13 MHz, DMSOd₆, CD₃CO₂D) δ 2.85 - 2.91 (4H, m), 3.75 (3H, s), 4.69 (2H, s), 6.02 (IH, s), 6.28 (IH, d), 6.73 - 6.76 (IH, m), 6.78 - 6.80 (2H, m), 7.17 (IH, t), 7.52 - 7.53 (IH, m), 7.86 (IH, d), 7.89 (IH, d). MS: m/z 445 (MH+) (FGFR Kinase assay - Caliper IC₅₀0.405μM)

2-Chloro-N-[5-[2-(3-methoxyphenyl)ethyl]-2H-pyrazol-3-yl]pyrimidin-4-amine used as starting material was prepared as follows:- 2,4-Dichloropyrimidine (2.65g, 17.79mmol, leq), 5-[2-(3-methoxyphenyl)ethyl]-2H- pyrazol-3 -amine (4.Og, 21.35mmol, 1.2eq) and N-ethyl-N-propan-2-yl-propan-2-amine (6.2mL, 35.58mmol, 2eq) were heated in ethanol (8OmL) at 5O⁰C for 5 days. The reaction was evaporated to dryness and the residue dissolved in ethyl acetate, washed with water, brine, dried (MgSO₄), filtered and evaporated to give crude product as a yellow oil. The oil was triturated with DCM to give a white solid which was filtered and dried on the filter giving the title compound (2.3 Ig, 39%).

IH NMR (DMSO 400.13MHz) 2.89 (4H, s), 3.73 (3H, s), 6.11 (IH, bs), 6.75 (IH, m), 6.81 (2H, m), 7.19 (IH, t), 8.15 (IH, d), 10.25 (IH, s) 12.18 (IH, s). MS m/z 330 (MH+).

5-[2-(3-Methoxyphenyl)ethyl]-2H-pyrazol-3-amine, used as starting material was prepared as in Example 2.

6-(Aminomethyl)pyridine-2-carboxamide, used as starting material was prepared as follows :- 6-[[(2-methylpropan-2-yl)oxycarbonylamino]methyl]pyridine-2-carboxylic acid (7.825g, 31.OOmmol, 1.OOeq) was dissolved in N,N-dimethylacetamide (32OmL) and ammonium chloride (14.885 g, 372.00mmol, 12.0eq) added, followed by N-ethyl-N-propan-2-yl- propan-2-amine (43.12 mL, 248. OOmmol, 8. OOeq) and 2-(7-Aza-lH-benzotriazole-l-yl)- 1,1,3,3-tetramethyluronium hexafluorophosphate (17.68g, 46.50mmol, 1.50eq). The mixture was stirred overnight at room temperature. Ammonium chloride was filtered off and the filtrate was concentrated in vacuo. The residue was partitioned between ethyl acetate (70OmL) and water (30OmL). The organic layer was washed successively with water (30OmL), saturated aqueous sodium bicarbonate solution (30OmL), brine (30OmL), then dried over magnesium sulphate and evaporated to give tert-butyl N- [(6- carbamoylpyridin-2-yl)methyl]carbamate as a clear oil, 7.54g (97% yield). This material was used in the next step without purification. tert-Butyl N-[(6-carbamoylpyridin-2-yl)methyl]carbamate (7.54g, 30.00mmol, l.OOeq) was placed in a 50OmL round bottomed flask and 2,2,2-trifluoroacetic acid (46.22mL, όOOmmol, 20.0eq) added. The solution became opaque quickly and after 5 minutes became a fine white suspension. Gas evolution and a slight exotherm (temperature rose to ~30°C) was noted. The suspension was stirred at room temperature for a further 10 mins. Ethyl acetate (25OmL) was added to give a clear solution. A solution of hydrogen chloride in diethyl ether (2.0M, 5OmL) was now added. This gave an immediate white precipitate. The suspension was stirred for 100 minutes and then filtered to give 6- (aminomethyl)pyridine-2-carboxamide hydrochloride as a white solid, 3.75g, (66.8% yield). 1H NMR (399.9 MHz, DMSO-d6) δ 4.28 (2H, d), 5.50 (5H, s), 7.64 - 7.69 (2H, m), 7.98 - 8.05 (2H, m), 8.73 (4H, d). MS: m/z 152 (MH+)

Example 8

6- [ [ [4- [ [5- [2-(3,5-dimethoxyphenyl)ethyl] -2H-pyr azol-3-yl] amino] pyrimidin-2- yl] amino] methyl] pyridine-2-carboxamide

A mixture 2-chloro-N-[5-[2-(3,5-dimethoxyphenyl)ethyl]-2H-pyrazol-3-yl]pyrimidin-4- amine (108mg, 0.30mmol, l.OOeq), 6-(aminomethyl)pyridine-2-carboxamide (107mg, 0.57mmol, 1.9eq), N-ethyl-N-propan-2-yl-propan-2-amine (183uL, 1.05mmol, 3.5eq) in 2- methoxyethanol (2mL) was heated in a microwave reactor at 180⁰C for 30 mins. The resulting solution was allowed to cool to room temperature and then evaporated to dryness. This material was purified by reverse-phase preparative HPLC (basic) using a 26-46% gradient of acetonitrile in water (containing 1% ammonium hydroxide) solution. The clean fractions were taken and evaporated to afford the title compound as a white solid, (lOlmg, 71% yield). IH NMR (500.13 MHz, DMSO-d6, CD3CO2D) δ 2.86 (4H, s), 3.72 (IH, s), 3.73 (6H, s), 4.69 (2H, s), 6.03 (IH, s), 6.29 (IH, d), 6.32 (IH, t), 6.39 (2H, d), 7.52 - 7.54 (IH, m), 7.85 - 7.91 (3H, m) (FGFR Kinase assay - Caliper IC₅₀0.2μM))

2-chloro-N-[5-[2-(3,5-dimethoxyphenyl)ethyl]-2H-pyrazol-3-yl]pyrimidin-4-amine, used as starting material was prepared as follows:- A solution of 5-[2-(3,5-dimethoxyphenyl)ethyl]-2H-pyrazol-3-amine (742 mg. 3.00 mmol, 1.2eq), 2,4-dichloropyrimidine (373mg, 2.5mmol, l.Oeq) and N-ethyl-N-propan-2-yl- propan-2-amine (871 μL, 5.0 mmol, 2eq) in ethanol (10 mL) was heated at 50⁰C for 36 h. The reaction mixture was concentrated and triturated with water to give a solid. The crude product was purified by silica column chromatography, eluting with 5% methanol in dichlorome thane. The desired fractions were combined and evaporated to afford 2-chloro- N-[5-[2-(3,5-dimethoxyphenyl)ethyl]-lH-pyrazol-3-yl]pyrimidin-4-amine as a white solid (401 mg, 47%). IH NMR (399.9 MHz, DMSOd₆) δ 2.87 (3H, t), 2.84 - 2.90 (IH, m), 3.72 (6H, s), 6.33 (IH, t), 6.42 (2H, d), 8.16 (IH, d), 10.28 (IH, s), 12.20 (IH, s). MS: m/z 360 (MH+).

5-[2-(3,5-dimethoxyphenyl)ethyl]-2H-pyrazol-3-amine, used as starting material was prepared as in Example 1.

6-(Aminomethyl)pyridine-2-carboxamide, used as starting material was prepared as in Example 7.

Example 9

6- [ [ [4- [ [5- [2-(4-methoxypyridin-2-yl)ethyl] -2H-pyr azol-3-yl] amino] pyrimidin-2- yl] amino] methyl] pyridine-2-carboxamide A mixture 2-chloro-N-[5-[2-(4-methoxypyridin-2-yl)ethyl]-2H-pyrazol-3-yl]pyrimidin-4- amine (58mg, O.lδmmol, l.Oeq), 6-(aminomethyl)pyridine-2-carboxamide hydrochloride (63mg, 0.32mmol, 1.78eq) and N-ethyl-N-propan-2-yl-propan-2-amine (107μL, 0.61mmol, 3.5eq) in 2-methoxyethanol (2mL) was heated in a microwave reactor at 180⁰C for 45 mins. The resulting solution was allowed to cool to room temperature and then evaporated to dryness. This material was purified by reverse-phase preparative HPLC (basic) using a 10-70% gradient of acetonitrile in water containing 5% ammonium hydroxide solution. The clean fractions were taken and evaporated to afford the title compound as a solid, (29.8mg, 37% yield).

¹H NMR (500.13 MHz, DMSOd₆, CD₃CO₂D) δ 2.95 - 2.99 (2H, m), 2.99 - 3.04 (2H, m), 3.82 (3H, s), 4.69 (2H, s), 6.02 (IH, s), 6.28 (IH, d), 6.75 - 6.78 (IH, m), 6.79 - 6.80 (IH, m), 7.52 - 7.54 (IH, m), 7.85 (IH, d), 7.87 - 7.91 (2H, m), 8.31 (IH, d). MS: m/z 446 (MH+) (FGFR Kinase assay - Caliper IC₅₀5.1 μM)

2-chloro-N-(5-(2-(4-methoxypyridin-2-yl)ethyl)-lH-pyrazol-3-yl)pyrimidin-4-amine, used as starting material was prepared as follows:-

N-Ethyl-N-propan-2-yl-propan-2-amine (0.555 mL, 3.21 mmol, 2eq) was added to 5-(2-(4- methoxypyridin-2-yl)ethyl)-lH-pyrazol-3 -amine (350mg, 1.60 mmol, leq) and 2,4- dichloropyrimidine (239 mg, 1.60 mmol, leq) in ethanol (5mL). The resulting solution was stirred at 50 ⁰C for 4 days. The reaction mixture was evaporated to dryness and redissolved in EtOAc (20 mL), and washed sequentially with water (20 mL) and saturated brine (20 mL). The organic layer was dried over MgSO4, filtered and evaporated to afford crude product. The crude product was purified by flash silica chromatography, elution gradient 50 to 100% EtOAc in isohexane then 10% MeOH in EtOAc. Pure fractions were evaporated to dryness to afford 2-chloro-N-(5-(2-(4-methoxypyridin-2-yl)ethyl)-lH- pyrazol-3-yl)pyrimidin-4-amine (174mg, 31.8 %) as a white solid.

IH NMR (400.132 MHz, DMSO) δ 3.02 (4H, s), 3.82 (3H, s), 6.28 (IH, s), 6.81 (IH, m), 6.86 (IH, d), 8.15 (IH, d), 8.31 (IH, d), 10.28 (IH, s), 12.25 (IH, s). MS: m/z 331 (MH+)

5-(2-(4-methoxypyridin-2-yl)ethyl)-lH-pyrazol-3-amine, used as starting material was prepared as follows:-

Acetonitrile (1.078 mL, 20.64 mmol, 1.3eq) was added dropwise to lithium diisopropylamide (10.32 mL, 20.64 mmol, 1.3eq) in THF (10OmL) at -78⁰C over a period of 5 mins under nitrogen. The resulting solution was stirred at -78 ⁰C for 10 mins. Methyl 3-(4-methoxypyridin-2-yl)propanoate (3.1g, 15.88 mmol, leq) was added in one portion and the reaction was stirred for 30 mins before being allowed to warm to ambient temperature. After stirring for 10 mins, a further 1.3 equivalents of acetonitrile anion was added to the reaction flask at -78 ⁰C. The reaction was allowed to warm back up to ambient temperature and stirred for 1 h. The reaction mixture was diluted with ethanol (100 mL) and hydrazine hydrochloride (2.83 g, 41.29 mmol, 2.6eq) was added. The reaction was heated at reflux for 3 h until formation of pyrazole was complete. The resulting mixture was evaporated to dryness and the residue was purified by flash silica chromatography, elution gradient 0 to 5% MeOH in DCM with ammonia. Pure fractions were evaporated to dryness to afford 5-(2-(4-methoxypyridin-2-yl)ethyl)-lH-pyrazol-3-amine (859mg, 24.78 %) as a yellow gum.

IH NMR (400.132 MHz, DMSO) δ 2.79 - 2.87 (2H, m), 2.87 - 2.96 (2H, m), 3.83 (3H, s), 6.73 (IH, s), 6.77 - 6.81 (IH, m), 6.83 (IH, d), 7.33 (IH, s), 8.30 (IH, d). MS: m/z 219 (MH+)

Methyl 3-(4-methoxypyridin-2-yl)propanoate, used as starting material was prepared as follows :- Methyl 3-(4-methoxypyridin-2-yl)prop-2-enoate (3.1g, 16.05 mmol, leq) and palladium on charcoal (310mg, 0.29 mmol, O.leq) in ethanol (20OmL) were stirred under an atmosphere of hydrogen at ambient temperature for 24 h. The reaction mixture was filtered through celite. The mixture was evaporated to dryness and the residue was azeotroped with Et₂O to afford desired methyl 3-(4-methoxypyridin-2-yl)propanoate (3.04 g, 15.56 mmol, 97 %). IH NMR (400.132 MHz, DMSO) δ 2.74 (2H, t), 2.95 (2H, t), 3.60 (3H, s), 3.82 (3H, s), 6.79 (IH, m), 6.86 (IH, d), 8.27 (IH, d). MS: m/z 196 (MH+).

Methyl 3-(4-methoxypyridin-2-yl)prop-2-enoate, used as starting material was prepared as follows :- Methyl 2-triphenylphosphoranylideneacetate (11.34 g, 33.91 mmol, 1.5eq) was added portionwise to 4-methoxypyridine-2-carbaldehyde (3.1g, 22.61 mmol, leq) in DCM (10OmL) at 2O⁰C over a period of 5 mins. The resulting solution was stirred at 20 ⁰C for 18 h. The resulting mixture was evaporated to dryness and the crude product was purified by flash silica chromatography, elution gradient 20 to 50% EtOAc in isohexane. Pure fractions were evaporated to dryness to afford methyl 3-(4-methoxypyridin-2-yl)prop-2- enoate (3.13 g, 16.18 mmol, 71.6 %) as a white solid. IH NMR (400.132 MHz, DMSO) δ 3.76 (3H, s), 3.88 (3H, s), 6.91 (IH, d), 7.00 (IH, m), 7.38 (IH, d), 7.63 (IH, d), 8.45 (IH, d). MS m/z 194 (MH+).

4-Methoxypyridine-2-carbaldehyde, used as starting material was prepared as follows:- Manganese(IV) oxide (2.412 mL, 139.42 mmol, 4eq) was added portionwise to (4- methoxypyridin-2-yl)methanol (4.85g, 34.85 mmol, leq) in ethyl acetate (15OmL). The resulting mixture was stirred at 80 ⁰C for 2 h. The hot reaction mixture was filtered through celite. The resulting mixture was evaporated to dryness to afford desired A- methoxypyridine-2-carbaldehyde (3.01 g, 21.93 mmol, 62.9 %). IH NMR (400.132 MHz, DMSO) δ 3.93 (3H, s), 7.27 (IH, m), 7.44 (IH, d), 8.63 (IH, d), 9.96 (IH, s)

(4-Methoxypyridin-2-yl)methanol, used as starting material was prepared as follows:- Trifiuoroacetic anhydride (50.8 mL, 359.32 mmol, 5eq) was added dropwise to A- methoxy-2-methylpyridine 1-oxide (1Og, 71.86 mmol, leq) in DCM (40OmL) at 0 ⁰C over a period of 20 mins under nitrogen. The resulting solution was stirred at room temperature for 30 mins. The temperature was increased to 40 ⁰C and the reaction mixture was stirred for a further 6 h then left to stir at room temperature overnight. The reaction mixture was quenched carefully with saturated sodium bicarbonate and stirred for 1 h. The reaction mixture was extracted with DCM and the organic layers were combined and washed with brine, dried (MgSO4), filtered and evaporated under reduced pressure to give (4- methoxypyridin-2-yl)methanol (5.56 g, 39.94mmol, 55.6 %) as a yellow oil. IH NMR (400.132 MHz, DMSO) δ 3.83 (3H, s), 4.52 (2H, s), 5.38 (IH, s), 6.82 (IH, m), 7.01 (IH, d), 8.29 (IH, d)

4-Methoxy-2-methylpyridine 1-oxide, used as starting material was prepared as follows:- 2-Methyl-4-nitropyridine 1-oxide (17.9g, 116.14mmol, leq) was added portionwise to a stirred solution of sodium methoxide (18.8g, 348mmol, 3eq) in methanol (550 mL). The mixture was heated at reflux for 3 h. The residue was dissolved in the minimum volume of water and neutralised with 2N HCl then extracted with DCM (x6). The organics were dried (MgSO₄), filtered and evaporated under reduced pressure to give the title compound as a yellow oil (15. Ig, 94%). ¹H NMR (400.13 MHz, DMSO) δ 2.34 (3H, s), 3.82 (3H, s), 6.90 (IH, m), 7.13 (IH, d), 8.12 (1H, d)

6-(Aminomethyl)pyridine-2-carboxamide, used as starting material was prepared as in Example 30.

Example 10

6-[ [ [4- [ [5- [2- [3-(methylcarbamoyl)phenyl] ethyl] -lH-pyrazol-3-yl] amino] pyrimidin-2- yl] amino] methyl] pyridine-2-carboxamide A mixture of 3-[2-[5-[(2-chloropyrimidin-4-yl)amino]-lH-pyrazol-3-yl]ethyl]-N-methyl- benzamide (83mg, 0.23mmol, l.Oeq), 6-(aminomethyl)pyridine-2-carboxamide (82mg, 0.41mmol, 1.78eq) and N-ethyl-N-propan-2-yl-propan-2-amine (141μL, O.δlmmol, 3.5eq) in 2-methoxyethanol (2mL) was heated in a microwave reactor at 180⁰C for 45 mins. The resulting solution was allowed to cool to room temperature and then evaporated to dryness. This material was purified by reverse-phase preparative HPLC (basic) using a 20-40% gradient of acetonitrile in water containing 5% ammonium hydroxide solution. The clean fractions were taken and evaporated to afford the title compound as a solid, (35mg, 32% yield). 1H NMR (500.13 MHz, DMSO-d6, CD3CO2D) δ 2.81 (3H, s), 2.88 - 2.91 (2H, m), 2.94 - 2.98 (2H, m), 4.69 (2H, s), 6.01 (IH, s), 6.27 (IH, d), 7.33 (2H, d), 7.52 - 7.53 (IH, m), 7.62 - 7.64 (IH, m), 7.71 (IH, s), 7.85 (IH, d), 7.86 - 7.91 (2H, m). MS: m/z 472 (MH+) (FGFR Kinase assay - Caliper IC₅₀0.079μM)

3-[2-[5-[(2-chloropyrimidin-4-yl)amino]-2H-pyrazol-3-yl]ethyl]-N-methyl-benzamide, used as starting material was prepared as follows :-

3-[2-(5-amino-lH-pyrazol-3-yl)ethyl]-N-methyl-benzamide (0.308g, 1.26mmol) was dissolved in ethanol (1OmL), then 2,4-dichloropyrimidine (0.157 g, 1.05mmol) was added followed by N-ethyl-N-propan-2-yl-propan-2-amine (0.366mL). The mixture was heated at 5O⁰C for 90 h, cooled, and evaporated to dryness to give an orange gum (0.5694g). Purified by flash silica chromatography, elution gradient 0-10% MeOH in DCM. Pure fractions were evaporated to dryness to afford 3-[2-[5-[(2-chloropyrimidin-4-yl)amino]-2H-pyrazol- 3-yl]ethyl]-N-methyl-benzamide as a yellow solid (0.3352g, 89%). IH NMR (399.9 MHz, DMSOd₆) δ 2.75 - 2.85 (3H, s), 2.85 - 3.05 (4H, m), 6.10 (IH, s), 7.40 (2H, m), 7.63 - 7.67 (IH, s), 7.73 - 7.78 (IH, s), 8.12 - 8.20 (IH, d), 8.35 - 8.42 (IH, s), 8.84 (IH, s), 10.24 - 10.33 (IH, s), 12.20 - 12.30 (IH, s) MS: m/z 357.00 (MH⁺)

3-[2-(5-amino-lH-pyrazol-3-yl)ethyl]-N-methyl-benzamide, used as starting material, was prepared as follows:- To a stirred suspension of 3-[2-(5-amino-lH-pyrazol-3-yl)ethyl]benzoic acid (1.62Og, 7.0mmol) and 2M N-methylmethanamine in THF (5.25mL, 10.5mmol) in dry DMF (5OmL), dry N-ethyl-N-propan-2-yl-propan-2-amine (4.63mL, 4eq, 28.0mmol) was added. O-(7-Azabenzotriazol- 1 -Yl)-N ,N,N',N'-Tetramethyluronium Hexafluoro-Phosphate (2.93g, 7.7mmol) was then added and the mixture left to stir for 18 hours. Evaporated to dryness, then partitioned between water (3OmL) and ethyl acetate (3OmL). Aqueous layer washed with ethyl acetate (3x30mL). Organics combined, washed sequentially with brine (lx30mL), 0.5N citric acid (lx30mL) and NaHCO₃ solution (lx30mL). Evaporated to dryness to afford crude 3-[2-(5-amino-lH-pyrazol-3-yl)ethyl]-N-methyl-benzamide as an orange gum (1.3594g). Purified by flash silica chromatography elution gradient 0-10% MeOH in DCM. Pure fractions evaporated to dryness to afford pure 3-[2-(5-amino-lH- pyrazol-3-yl)ethyl]-N-methyl-benzamide (0.33Og, 28%).

IH NMR (399.9 MHz, DMSO-d₆) δ 2.74 - 2.79 (2H, m), 2.76 - 2.78 (3H, m), 2.89 (2H, d), 3.20 - 3.45 (2H, s), 5.21 (IH, s), 7.35 - 7.36 (2H, m), 7.63 - 7.66 (IH, m), 7.72 (IH, s), 8.36 - 8.37 (IH, m) MS: m/z 245.41 (MH⁺)

3-[2-(5-Amino-lH-pyrazol-3-yl)ethyl]benzoic acid used as starting material,was prepared as follows :-

A suspension of 3-[2-(5-amino-2H-pyrazol-3-yl)ethyl]benzonitrile (4.00Og, 19.0mmol) in an aqueous solution of sodium hydroxide (1OM, 4OmL) was heated at 95-100⁰C for 5 h. The reaction mixture was cooled to 5-1O⁰C in an ice/water bath and acidified to pH3 by the dropwise addition of cone. HCl (approx. 5OmL). The resultant cream solid was removed by filtration, washed with water and then dried in a vacuum oven over the weekend to leave pure 3-[2-(5-amino-lH-pyrazol-3-yl)ethyl]benzoic acid (4.4208g, 101% yield). ¹H NMR (399.9 MHz, DMSOd₆) 52.79 (2H, d), 2.95 (2H, d), 5.29 (IH, s), 7.41 (IH, t), 7.48 (IH, d), 7.77 (IH, s), 7.79 (IH, s), 7.82 (IH, d) MS: m/z 232.39 (MH⁺)

3-[2-(5-amino-2H-pyrazol-3-yl)ethyl]benzonitrile, used as starting material,was prepared as follows :-

Sodium hydride (60%, 3.Og, 75.6mmol) was added to a stirred solution of methyl 3-(3- cyanophenyl)propanoate (11.9g, 63.0mmol) in dry 1,4 dioxane (35OmL) and dry acetonitrile (3.95mL, 75.6mmol) under nitrogen to give a cloudy grey mixture. This was stirred at room temperature for 10 mins and then refluxed under nitrogen overnight to give a dark orange solution. The reaction mixture was cooled and ethanol (25mL) was added followed by hydrazine monohydrochloride (8.635g, 126mmol). The reaction mixture was refluxed overnight. The reaction mixture was cooled, filtered, and evaporated to dryness to afford crude 3-[2-(5-amino-2H-pyrazol-3-yl)ethyl]benzonitrile (16g). Purified by flash silica chromatography, eluted isocratically with 8% MeOH in DCM. Pure fractions evaporated to dryness to afford 3-[2-(5-amino-2H-pyrazol-3-yl)ethyl]benzonitrile as an orange gum, (5.1g, 38%). IH NMR (399.9 MHz, DMSOd₆) δ 2.73 - 2.76 (2H, m), 2.88 - 2.92 (2H, m), 4.07 - 4.08 (IH, m), 4.50 (2H, s), 5.17 (IH, s), 7.47 - 7.51 (IH, m), 7.55 - 7.58 (IH, m), 7.64 - 7.66 (2H, m) MS: m/z 213.41 (MH⁺)

Methyl 3-(3-cyanophenyl)propanoate, used as starting material,was prepared as follows:- To a solution of methyl (E)-3-(3-cyanophenyl)prop-2-enoate (12.36g, 66.00mmol) dissolved in DMF (25OmL), was added platinum catalyst (1.24g) and the reaction mixture was stirred under hydrogen overnight. The mixture was filtered through celite, washed with DMF, then evaporated to dryness to give a grey-brown liquid. The solid was dissolved in DCM (15OmL) and washed sequentially with water (3x80mL) and brine (lx80mL), then dried with MgSO₄, and evaporated to dryness to afford methyl 3-(3- cyanophenyl)propanoate as a brown liquid (11.949g, 96%). ¹H NMR (399.9 MHz, DMSOd₆) 52.69 (2H, t), 2.90 - 2.94 (2H, m), 3.59 (3H, s), 7.50 (IH, t), 7.60 - 7.62 (IH, m), 7.66 - 7.69 (IH, m), 7.73 (IH, d)

Methyl (E)-3-(3-cyanophenyl)prop-2-enoate, used as starting material,was prepared as follows :-

Methyl (triphenyphosphoranylidene)acetate (38.12g, 114mmol) was added to a mixture of 3-cyanobenzaldehyde (9.97g, 76mmol) in DCM (15OmL) and the reaction mixture was stirred for 6 h at room temperature. The reaction mixture was evaporated to dryness to afford crude methyl (E)-3-(3-cyanophenyl)prop-2-enoate. Purified by flash silica chromatography, eluted isocratically with 50% ethyl acetate in isohexanes. Pure fractions evaporated to dryness to afford pure methyl (E)-3-(3-cyanophenyl)prop-2-enoate (12.36g, 87%).

¹H NMR (399.9 MHz, DMSOd₆) 53.76 (3H, s), 6.84 (IH, s), 7.64 (IH, t), 7.68 (IH, s), 7.87 - 7.89 (IH, m), 8.06 - 8.09 (IH, m), 8.27 (IH, t)

Example 11 6- [ [ [4- [ [5- [2-(3-hydroxyphenyl)ethyl] -2H-pyr azol-3-yl] amino] pyrimidin-2- yl] amino] methyl] pyridine-2-carboxamide

6-({[4-({3-[2-(3-Methoxyphenyl)ethyl]-lH-pyrazol-5-yl}amino)pyrimidin-2- yl] amino }methyl)pyridine-2-carboxamide (147mg, 0.33mmol, l.Oeq) was dissolved in dichloromethane (15mL) and cooled to O⁰C under an atmosphere of nitrogen. Boron tribromide solution (1.0M in dichloromethane, 1.65mL , 1.65mmol, 5.0eq) was added dropwise and the reaction was then allowed to warm to room temperature and stirred for 18 h. The mixture was cooled in an ice bath and quenched carefully with methanol (1OmL) and the solution was evaporated to dryness. The residue was partitioned between ethyl acetate (5OmL) and sodium bicarbonate (25mL) solution. 1-Butanol was added to aid solubility. The organic layer was washed with water and then allowed to separate overnight. It was then evaporated to give a pale brown solid, 181mg. This material was purified by reverse-phase preparative HPLC (basic) using a 25-85% gradient of acetonitrile in water containing 5% ammonium hydroxide solution. The clean fractions were taken and evaporated to afford the title compound as a white solid, (30.9mg, 21% yield). 1H NMR (500.13 MHz, DMSOd₆, CD₃CO₂D) δ 2.83 (4H, s), 4.69 (2H, s), 6.03 (IH, s), 6.29 (IH, d), 6.59 - 6.61 (IH, m), 6.64 - 6.66 (2H, m), 7.04 - 7.07 (IH, m), 7.52 - 7.54 (IH, m), 7.85 (IH, d), 7.86 - 7.91 (2H, m). MS: m/z 431 (MH+). (FGFR Kinase assay - Caliper IC₅₀0.13μM)

6-({[4-({3-[2-(3-Methoxyphenyl)ethyl]-lH-pyrazol-5-yl}amino)pyrimidin-2- yl] amino }methyl)pyridine-2-carboxamide was prepared as in Example 7.

Example 12

(S)-3-(2-(5-(2-(l-(5-fluoropyridin-2-yl)ethylamino)pyrimidin-4-ylamino)-lH-pyrazol-

3-yl)ethyl)benzonitrile

A mixture of (S)-4-chloro-N-(l-(5-fluoropyridin-2-yl)ethyl)pyrimidin-2-amine (292mg, l.lόmmol), 3-(2-(5-amino-lH-pyrazol-3-yl)ethyl)benzonitrile (294mg, 1.39mmol), hydrogen chloride (0.318mL of a 4M solution in dioxane, 1.27mmol) in dioxane (1OmL) was heated at 150⁰C for 1 hour under microwave irradiation. The mixture allowed to cool, loaded on to an SCX-2 ion-exchange column, non-basic impurities were eluted with methanol and the product eluted with 7N methanolic ammonia. The solvent was removed from product containing fractions by evaporation. The residue (434mg) was purified by reverse-phase preparative HPLC (basic) using a 26-46% gradient of acetonitrile in water

(containing 1% ammonium hydroxide) solution to give the title compound (131 mg,

26.5%).

IH NMR (500 MHz, DMSO/CD3COOD @ 373K): δH :1.49 (d, 3H), 2.90 (t, 2H), 3.01 (t, 2H), 5.18 (q, IH), 5.97 (s, IH), 6.19 (d, IH), 7.45 (m, 2H), 7.55 (m, 3H), 7.61 (s, IH), 7.80

(d, IH), 8.42 (s, IH) ; MS m/z 429 (M+H)+ .

(FGFR Kinase assay - Caliper IC₅₀0.25 μM)

3-(2-(5-amino-lH-pyrazol-3-yl)ethyl)benzonitrile was prepared as described in Example

48 of WO2008/001070. (S)-4-chloro-N-(l-(5-fluoropyridin-2-yl)ethyl)pyrimidin-2-amine was prepared as follows:

Saturated aqueous potassium carbonate solution (100ml) was added to (S)-l-(5- fluoropyridin-2-yl)ethanamine dihydro chloride (5g, 23.47mmol). Th aqueous mixture was extracted with DCM (3 x 100ml), the organics were combined, dried (Na₂SO₄) and the solvent removed by evaporation. Ethyl formate (3OmL) was added to the resulting oil and the mixture heated at 6O⁰C for 16 hours. The volatiles were removed by evaporation to give (S)-N-(I -(5 -fluoropyridin-2-yl)ethyl)formamide (2.91 g, 73.7 %) as a brown oil; IH NMR (300.132 MHz, DMSO) δ 1.37 (3H, d), 5.05 (IH, quintet), 7.43 (IH, m), 7.70 (IH, m), 8.05 (IH, s), 8.51 (IH, d), 8.56 (IH, s); MS m/z 169 (M+H)+ Sodium hydride (60% in mineral oil) (0.724g, 18.1 lmmol) was added to a stirred solution of (S)-N-(I -(5 -fiuoropyridin-2-yl)ethyl)formamide (2.9g, 17.24mmol) in THF (10OmL) cooled to 0 ⁰C. The mixture was stirred for 30 minutes and then 4-chloro-2- (methylsulfonyl)-pyrimidine (3.32g, 17.24mmol) was added and the mixture stirred for 24 hours at 0 ⁰C. The solvent was removed by evaporation from the mixture without heating. Iced water added was added to the residue followed by 2N aqueous sodium hydroxide (to remove formyl group) and the mixture immediately extracted with DCM, The extracts were combined, washed with saturated aqueous sodium chloride solution and dried (Na₂SO₄). The solvent was removed by evaporation and the residue purified by column chromatography on silica gel eluting with iso-hexane followed by isohexane / dichloromethane (1:1) followed by dichlorome thane and finally diethylether / dichloromethane (increasing polar mixtures 1:3 to 1:1) to give (S)-4-chloro-N-(l-(5- fiuoropyridin-2-yl)ethyl)pyrimidin-2-amine (2.98g, 68.4%); IH NMR (399.9 MHz, DMSO): δH :1.82 (d, 3H), 6.04 (m, IH), 7.40 (d, IH), 7.49 (m, IH), 7.65 (m, IH), 8.41 (s, IH), 8.60 (d, IH), 9.68 (s, IH); MS m/z 253 (M+H)+

Example 13

(S)-N2-(l-(5-fluoropyridin-2-yl)ethyl)-N4-(3-(2-(pyridin-3-yl)ethyl)-lH-pyrazol-5- yl)pyrimidine-2,4-diamine

A mixture of (S)-4-chloro-N-(l-(5-fluoropyridin-2-yl)ethyl)pyrimidin-2-amine (251mg, 0.99mmol), 3-(2-(pyridin-3-yl)ethyl)-lH-pyrazol-5-amine (224mg, 1.19mmol), hydrogen chloride (0.323mL of a 4M solution in dioxane , 1.29mmol) in dioxane (1OmL) was heated at 150⁰C for 4 hour under microwave irradiation. The mixture allowed to cool, loaded on to an SCX-2 ion-exchange column, non-basic impurities were eluted with methanol and the product eluted with 7N methanolic ammonia. The solvent was removed from product containing fractions by evaporation. The residue was purified by reverse- phase preparative HPLC (basic) using a 26-46% gradient of acetonitrile in water

(containing 1% ammonium hydroxide) solution to give the title compound (24.40mg, 6.07%)

IH NMR (500 MHz, DMSO/CD3COOD @ 373K): δH :1.50 (d, 3H), 2.89 (m, 2H), 2.96 (m, 2H), 5.19 (q, IH), 5.96 (s, IH), 6.19 (d, IH), 7.25 (m, IH), 7.45 (m, IH), 7.54 (m, IH), 7.59 (m, IH), 7.80 (d, IH), 8.37 (m, IH), 8.43 (m, 2H); MS m/z 405 (M+H)+

(FGFR Kinase assay - Caliper IC₅₀0.53μM)

3-(2-(pyridin-3-yl)ethyl)-lH-pyrazol-5-amine was prepared as described in Example 7 of WO2008/075068.

Example 14

(S)-N2-(l-(5-fluoropyridin-2-yl)ethyl)-N4-(3-(3-methoxyphenethyl)-lH-pyrazol-5- yl)pyrimidine-2,4-diamine

5-Bromo-2-chloro-N-(3-(3-methoxyphenethyl)-lH-pyrazol-5-yl)pyrimidin-4-amine (0.5g, 1.22mmol), (S)-l-(5-fluoropyridin-2-yl)ethanamine, dihyrochloride (0.287g, 1.35mmol), and N-ethyldiisopropylamine (0.74ImL, 4.28mmol) dissolved in NMP (4mL). The solution was heated at 175°C for 2 hour under microwave irradiation. The mixture allowed to cool, loaded on to an SCX-2 ion-exchange column, non-basic impurities were eluted with methanol and the product eluted with 7N methanolic ammonia. The solvent was removed from product containing fractions by evaporation to give (S)-5-bromo-N2- ( 1 -(5 -fluoropyridin-2-yl)ethyl)-N4-(3 -(3 -methoxyphenethyl)- 1 H-pyrazol-5 -yl)pyrimidine- 2,4-diamine. Ammonium formate (231mg, 3.66mmol) and 10% palladium on charcoal catalyst (57.0mg, 0.54mmol) in ethanol (1OmL) was added this product (which was used without further purification) and the mixture heated at reflux for 4 hours. The mixture was allowed to cool, insolubles removed by filtration through diatomeous earth and the pad washed through with ethanol. The solution was loaded on to a SCX-2 ion exchange column, non-basic impurities were eluted with methanol and the product eluted with 7N methanolic ammonia. The solvent was removed from product containing fractions by evaporation. The residue was purified by column chromatography on silica gel eluting with methanol / dichloromethane (increasingly polar mixtures 0: 100 to 10:90) to give the title compound (228 mg, 43.1 %) as a beige foam.

IH NMR δH(300 MHz, DMSO-d6) 1.46 (3H d, J= 7.0 Hz), 2.82 - 2.90 (4H, m), 3.73 (3H, s), 5.08 - 5.18 (IH, m), 6.17 (IH, s), 6.74 - 6.83 (3H, m), 7.20 (IH t, J= 8.0 Hz), 7.45 (IH q, J= 4.4 Hz), 7.61 - 7.68 (IH, m), 7.78 (IH d, J= 5.7 Hz), 8.46 (IH d, J= 3.3 Hz), 9.30 (IH, s), 11.88 (IH, s). MS m/z 434 (M+H)+

(FGFR Kinase assay - Caliper IC₅₀ 0.0042μM)

5-bromo-2-chloro-N-(3-(3-methoxyphenethyl)-lH-pyrazol-5-yl)pyrimidin-4-amine was prepared as follows:-

A mixture of 5-bromo-2,4-dichloropyrimidine (4.91g, 21.55mmol), 3-(3- methoxyphenethyl)-lH-pyrazol-5 -amine (4.68g, 21.55mmol),N-ethyldiisopropylamine (4.85mL, 28.01mmol) in THF (5OmL) was heated at reflux for 2 hours under nitrogen. The mixture was allowed to cool and the volatiles removed by evaporation. The residue was trituated with water, the resulting solid collected by filtration, washed with water and then diethylether and dried to. give 5-bromo-2-chloro-N-(3-(3-methoxyphenethyl)-lH- pyrazol-5-yl)pyrimidin-4-amine (6.84g, 78%) as a white solid. IH NMR δH(300 MHz, DMSO-d6) 2.49-2.52 (4H, m), 3.73 (3H, s), 6.30 (IH, s), 6.74 - 6.78 (IH, m), 6.81 - 6.84 (2H, m), 7.17 - 7.22 (IH, m), 8.42 (IH, s), 9.24 (IH, s), 12.38 (IH, s); MS m/z 410 (M+H)+

3-(3-methoxyphenethyl)-lH-pyrazol-5-amine was prepared as described in Example 2.

Kinase assays

FGFR Kinase assay - Elisa

To determine inhibition of FGFR activity, kinase assays were conducted using ELISA (Enzyme-Linked Immunosorbent Assay) technology. Kinase activity assays were performed in 384-well polypropylene plates (Matrix, 4311) with a total volume of 40μl in each well. Each well was coated with 2μg of polyEAY substrate (Sigma, P3899) at 4°C overnight. The plates were then washed once with lOOμl PBS and once with lOOμl 5OmM HEPES (pH 7.4) prior to the addition of the kinase assay reagents. Each kinase reaction contained 0.1 ng His₆-tagged FGFR kinase domain (FGFR kinase domain (amino acids 458-765, C488A, C584S) N-terminally fused to a His6-tag and TEV cleavage site encoded by the following sequence; [MHHHHHHEFKGSTSLYKKAGSSENLYFQGA]. The final alanine denotes the start of the FGFR protein sequence. The resultant protein was expressed and purified based on Mohammadi et al, Cell VoI 86, 577-587 (1996)), 5OmM HEPES (pH 7.4), O.lmM Na₃VO₄, O.lmM DTT, 0.05% (v/v) Triton XlOO, 2OmM MgCl₂, 160μM ATP. Various concentrations of test compounds were each added in 5% (v/v) DMSO to yield a final assay DMSO concentration of 1.25% (v/v). The kinase reactions were incubated at room temperature for 45 minutes and stopped by washing the plate three times with lOOμl PBS plus 0.05% Tween. 40μl of a one in 10000 dilution of 4G10-HRP antibody (Upstate Biotechnology, UBI 16-105) made up in 0.5% (w/v) BSA/ PBS was then added to each well and the plates incubated at room temperature for one hour. Following this, the plates were then washed repeatedly with lOOμl PBS plus 0.05% Tween to remove all traces of the antibody solution. 40μl of 50μg/mL 3,3',5,5'-Tetramethylbenzidine (Sigma, T2885), 0.05M phosphate-citrate buffer, containing 0.03% sodium perborate was added to each well and the plates incubated at room temperature for twelve minutes. The colour reaction was stopped by the addition of 20μl 2M H₂SO₄ and the plates read at 450nm on a Spectrafluor Plus (Tecan).

In the assay, compounds were tested at a range of concentrations. The mean data values for each concentration, along with untreated control wells and 100% inhibition control wells were used to derive a plot of inhibition against concentration. From this data, the IC50 value or a percentage inhibition value at fixed concentration may be determined.

Data may also be presented as a Percentage inhibition at IuM, this is a calculated value based on the curve fit that was generated experimentally. From the fitted curve plot, the effect of compound at a concentration of IuM was calculated as a percentage inhibition. A percentage inhibition calculated in this way from the shape of the curve is considered to be statistically more accurate compared to a single raw data value measured at a single concentration.

FGFR Kinase assay - Caliper

To determine inhibition of FGFR activity, kinase assays were conducted using Caliper technology.

Kinase activity assays were performed in Greiner 384-well low volume plates, with a total reaction volume of 12ul per well. Final concentration of FGFRl active kinase in each reaction well was 7.2nM. The substrate for each assay was a custom peptide with fluorescent tag (13 amino acids in length, 5FAM-KKSRGDYMTMQIG-CONH₂) the sequence of which was specific for FGFRl kinase.

Compounds were serially diluted in 5% (v/v) DMSO, before being added to assay plates. The Enzyme (at 7.2nM [final]) and Substrate (at 3.6uM [final]) were added separately to the compound plates, in reaction buffer [comprising: 5OmM MOPS - pH 6.5,

0.004% Triton, 2.4mM DTT, 12mM MgCl₂, 408uM ATP] resulting in a final DMSO concentration in the reaction mix of 0.8%.

Assay plates were incubated at room temperature for 1.5h, before the reaction was stopped with the addition of buffer [comprising: 10OmM HEPES - pH7.5, 0.033% Brij-35, 0.22% Caliper Coating Reagent #3, 88mM EDTA, 5% DMSO]. Stopped assay plates were then read using the Caliper LabChip® LC3000 (which uses micro fludics to measure a shift in mobility between fluorescent labelled peptide and the FGFRl kinase - phosphorylated form of this peptide).

Inhibition may also be expressed as Percentage inhibition at IuM, which is a calculated value based on the curve fit that was generated experimentally. From the fitted curve plot, the effect of compound at a concentration of IuM was calculated as a percentage inhibition. A percentage inhibition calculated in this way from the shape of the curve is considered to be statistically more accurate compared to a single raw data value measured at a single concentration. Similarly, inhibition activity of the other family members may also be evaluated, for example FGFR4 inhibition activity may be measured as follows:-

FGFR4 Kinase assay - Caliper

To determine inhibition of FGFR4 activity, kinase assays were conducted using Caliper technology.

Kinase activity assays were performed in Greiner 384-well low volume plates, with a total reaction volume of 12ul per well. Final concentration of FGFR4 active kinase in each reaction well was 25nM. The substrate for each assay was a custom peptide with fluorescent tag (13 amino acids in length, 5FAM-EEPLYWSFPAKKK-CONH₂) the sequence of which was specific for FGFR4 kinase.

Compounds were serially diluted in 5% (v/v) DMSO, before being added to assay plates. The Enzyme (at 25nM [final]) and Substrate (at 1.5uM [final]) were added separately to the compound plates, in reaction buffer (comprising: 10OmM HEPES - pH

7.5, 0.004% Triton, ImM DTT[fmal], 1OmM MnCl₂[fmal], 3OuM ATP[fmal]) resulting in a final DMSO concentration in the reaction mix of 0.8%.

Assay plates were incubated at room temperature for 2hrs, before the reaction was stopped with the addition of buffer [comprising: 10OmM HEPES - pH7.5, 0.033% Brij-35, 0.22% Caliper Coating Reagent #3, 4OmM EDTA, 5% DMSO]. Stopped assay plates were then read using the Caliper LabChip® LC3000 (which uses micro fludics to measure a shift in mobility between fluorescent labelled peptide and the FGFR4 kinase - phosphorylated form of this peptide).

Percentage inhibition at IuM, as expressed herein, is a calculated value based on the curve fit that was generated experimentally. From the fitted curve plot, the effect of compound at a concentration of IuM was calculated as a percentage inhibition. A percentage inhibition calculated in this way from the shape of the curve is considered to be statistically more accurate compared to a single raw data value measured at a single concentration.

Selective inhibition of the FGFR family members or inhibition of one or more FGFR family members may offer advantage in producing an anti-cancer effect mediated alone or in part by the inhibition of one or more FGFR's.

Trk A kinase inhibition

Trk A kinase activity was determined by measuring the kinase's ability to phosphorylate synthetic tyrosine residues within a generic polypeptide substrate using an Amplified Luminescent Proximity Assay (Alphascreen) technology (PerkinElmer, 549 Albany Street, Boston, MA).

To measure Trk A kinase activity, the intracellular domain of a HIS-tagged human Trk A kinase (amino acids 442-796 of Trk A, Swiss-Prot Primary Accession Number P04629) was expressed in SF9 cells and purified using standard nickel column chromatography. After incubation of the kinase with a biotinylated substrate and adenosine triphosphate (ATP) for 60 minutes at room temperature, the kinase reaction was stopped by the addition of 30 mM ethylenediaminetetraacetic acid (EDTA). The reaction was performed in 384 well low volume microtitre plates and the reaction products were detected with the addition of streptavidin coated Donor Beads and phosphotyrosine-specific antibodies coated Acceptor Beads using the EnVision Multilabel Plate Reader after an overnight incubation at room temperature.

The three TRK receptor isoforms are widely expressed in neuronal tissue during development. Compounds which display selectivity for FGFR over TRK may be desirable as they may show lessoned effects on certain tissues. This may amelerioate the potential for toxicity risks and may deliver therapeutic margin benefits.

Cell Assays

Cell based inhibition of transiently expressed FGFRl IHc phosphorylation. Measured using phospho-specific primary and fluorescent secondary antibodies.

This assay is designed to detect inhibitors of transiently expressed FGFRl phosphorylation by antibody staining of fixed cells detected using ArrayScan technology.

Cos-1 cells were routinely passaged in DMEM (Gibco BRL, 41966) plus 3% foetal calf serum (FCS), 1% L-glutamine (Gibco BRL, 25030) to a confluence of 80%. To undertake the assay, Cos-1 cells were harvested at 90-95% confluence for cell transfection.

For each 96-well plate, 24ul Lipofectamine 2000 was added to 809ul OptiMEM and incubated at room temperature for 5 minutes. For each 96 well plate, 20ug 3' FLAG tagged FGFRl/ pcDNA3.1 (In-house clonel5, MSD 4793) was diluted with OptiMEM to a total volume of 833ul. Equal volumes of DNA and Lipofectamine 2000 were combined (DNA: Lipid = 1:1.2 ratio) and incubated at room temperature for 20 minutes.

The harvested Cos-1 cells are counted using a coulter counter and diluted further with 1% FCS/DMEM to 2.5 x 10⁵ cells/mL. For each 96-well, 8.33mL cells were required. The complexed transfection solution was added to the cell solution and the cells were seeded at 2.5xlO⁵ cells/ well in DMEM plus 1% foetal calf serum, 1% L-glutamine in 96 well plates (Costar, 3904) and incubated at 37°C (+5% CO₂) in a humidified incubator overnight (24hrs). The following day, the plates were dosed with 25 ul compound (diluted from 10 mM stock in DMSO using serum free DMEM) and the plates were returned to a humidified 37°C (+5% CO₂) incubator for one hour. Media was removed from the wells using vacuum aspiration; cells were fixed by adding 50ul of 100% methanol to each well and incubated at room temperature for 20 minutes. The fixative solution was then removed and the wells were washed once with 200ul phosphate buffered saline (PBS/A) before permeabilising the cells by the addition of 50ul/ well 0.1% triton/ PBS/A for 20 minutes at room temperature. The permeabilisation solution was then removed and the cells washed once more with 200ul / well PBS/A before the addition of 40ul 1/1000 primary antibody solution (Cell Signalling Technologies #CS3476; mouse anti-phospho FGFRl diluted in PBS/A with 10% FCS + 0.1% Tween20) to each well.

Following incubation at room temperature for 1 hour, the antibody solution was removed and the wells were washed once with 200ul / well PBS/A. Then 40ul 1/500 secondary antibody (Al 1005; goat anti-mouse 594) solution and 1/10000 Hoechst (diluted together in PBS/A with 10% FCS + 0.1% Tween 20) were added and the plate incubated in the dark at room temperature for one hour. Finally, the plates were washed once with 200ul / well PBS/A, leaving the final wash in the wells before sealing the plates. The plates were read on an Arrayscan (Cellomics). The Channel 2 (594nm) values obtained from undosed (max) and reference compound (min) wells within a plate are used to set boundaries for 0% and 100% compound inhibition. Compound data is normalized against these values to determine the dilution range of a test compound that gives 50% inhibition of phosphorylated FGFRl .

Growth factor stimulated Erk phosphorylation

These and other assays were used to evaluate the ability of a test compound to inhibit growth factor stimulated cellular signalling in mammalian cell lines. This was achieved by measuring the amount of receptor tyrosine kinase regulated Erk phosphorylation within a cell following compound treatment. NIH 3T3 (ECACC, 93061524) cells were routinely passaged in DMEM (Gibco

BRL, 41966) plus 10% foetal calf serum (FCS), 1% L-glutamine (Gibco BRL, 25030) to a confluence not greater than 80%. To undertake the assay, NIH 3T3's were seeded at IxIO⁴ cells/ well in DMEM plus 10% foetal calf serum, 1% L-glutamine in 96 well plates (Costar, 3904) and incubated at 37°C (+5% CO₂) in a humidified incubator. Once the cells had fully adhered (typically following 4-5 hours incubation) the media was removed from each well and the cells gently washed with lOOμl warm serum free media. 90μl of serum free DMEM plus 1% L-glutamine was then added to each well and the plates were returned to a humidified 37°C (+5% CO₂) incubator. The following day, the plates were dosed with lOμl compound (diluted from 10 mM stock in DMSO using serum free DMEM) and the plates were returned to a humidified 37°C (+5% CO₂) incubator for one hour. NIH 3T3 cells were then stimulated with a final concentration of 3 ng/mL bFGF (Sigma, F0291) for 20 minutes at 37°C. Following stimulation the cells were fixed by adding formaldehyde (4% v/v final concentration) and incubating at room temperature for 20 minutes. The fixative solution was then removed and the wells were washed twice with lOOμl phosphate buffered saline (PBS/ A) before permeabilising the cells by the addition of 50μl/ well 0.1% triton/ PBS/A for 10 minutes at room temperature. The permeabilisation solution was then removed and the cells washed twice more with lOOμl/ well PBS/A before the addition of 50μl/ well anti-phospho p44/42 (Cell Signalling Technology, 9106), diluted 1/500 with PBS/A plus 10% FCS. The anti-phospho p44/42 antibody recognises Erk phosphorylated at threonine 202 and tyrosine 204. Following incubation at room temperature for 2 hours, the antibody solution was removed and the wells were washed twice with lOOμl/ well PBS/A. 50μl/ well 1/250 goat anti-mouse alexa fluor 488 secondary antibody (Molecular Probes, AI lOOl) and 1/10000 Hoescht (Molecular Probes, H-3570) diluted with PBS/A plus 10% FCS was added and the plate incubated in the dark at room temperature for one hour. Finally, the plates were washed three times with lOOμl/ well PBS/A, leaving the final wash in the wells before sealing the plates. The plates were read at 350nm and 488nm using an Arrayscan (Cellomics). The mean average intensity fluorescence values for each test compound concentration, untreated control wells and 100% inhibition control wells were used to determine the test compounds IC50 value. IC50 value is the concentration of test compound that inhibits 50% of Erk phosphorylation.

Cell based inhibition of transiently expressed FGFRl IHc phosphorylation via use of ECHO technology (measured using phospho-specific primary and fluorescent secondary antibodies).

This assay is designed to detect inhibitors of transiently expressed FGFRl phosphorylation by antibody staining of fixed cells detected using ArrayScan technology. Cos-1 cells were routinely passaged in DMEM (Gibco BRL, 41966) plus 3% foetal calf serum (FCS), 1% L-glutamine (Gibco BRL, 25030) to a confluence of 80%. To undertake the assay, Cos-1 cells were harvested at 90-95% confluence for cell transfection. For each 96-well plate, 24μl Lipofectamine 2000 was added to 809ul OptiMEM and incubated at room temperature for 5 minutes. For each 96 well plate, 20ug 3' FLAG tagged FGFRl/ pcDNA3.1 (In-house clonel5, MSD 4793) was diluted with OptiMEM to a total volume of 833μl. Equal volumes of DNA and Lipofectamine 2000 were combined (DNA: Lipid = 1:1.2 ratio) and incubated at room temperature for 20 minutes. The harvested Cos-1 cells are counted using a coulter counter and diluted further with 1% FCS/DMEM to 2.5 x 10⁵ cells/ml. For each 96-well, 8.33ml cells were required. The complexed transfection solution was added to the cell solution and the cells were seeded at 2.5xlO⁵ cells/ well in DMEM plus 1% foetal calf serum, 1% L-glutamine in 96 well plates (Costar, 3904) and incubated at 37°C (+5% CO₂) in a humidified incubator overnight (24hrs).

The following day,_compounds from dry weight samples were dissolved in 100% DMSO to give 1OmM concentration. 40μl of the compound was dispensed into the wells of each quadrant across the 384 Labcyte plate (inclusive of a positive control (100% DMSO), a negative control (lOμM) and a reference compound (25OnM)). The 384 Labcyte plate was then transferred to the Hydra to dilute the compounds 1:100 into the remaining wells of the quadrant. 70μl of media was aspirated from the assay plate using the Quadra before the plate was transferred onto the ECHO 550. The 384 Labcyte compound plate was also transferred onto the ECHO 550. Compound transfer to the assay plate on the ECHO 550 was at concentration ranges 1) 10 μM, 2) 3 μM, 3) 1 μM, 4) 0.3 μM, 5) 0.1 μM, 6) 0.01 μM.

The plates were gently tapped to mix compound in with the cell media and left to incubate at 37⁰C with 5% CO₂ for 1 hour.

Media was removed from the wells using vacuum aspiration; cells were fixed by adding 50μl of 100% methanol to each well and incubated at room temperature for 20 minutes. The fixative solution was then removed and the wells were washed once with 200μl phosphate buffered saline (PBS/A) before permeabilising the cells by the addition of 50μl/ well 0.1% triton/ PBS/A for 20 minutes at room temperature. The permeabilisation solution was then removed and the cells washed once more with 200μl / well PBS/A before the addition of 40μl 1/1000 primary antibody solution (Cell Signalling Technologies

#CS3476; mouse anti-phospho FGFRl diluted in PBS/A with 10% FCS + 0.1% Tween20) to each well.

Following incubation at room temperature for 1 hour, the antibody solution was removed and the wells were washed once with 200μl / well PBS/A. Then 40μl 1/500 secondary antibody (Al 1005; goat anti-mouse 594) solution and 1/10000 Hoechst (diluted together in PBS/A with 10% FCS + 0.1% Tween 20) were added and the plate incubated in the dark at room temperature for one hour. Finally, the plates were washed once with 200μl / well PBS/A, leaving the final wash in the wells before sealing the plates. The plates were read on an Arrayscan (Cellomics). The Channel 2 (594nm) values obtained from undosed (max) and reference compound (min) wells within a plate are used to set boundaries for 0% and 100% compound inhibition. Compound data was normalized against these values to determine the dilution range of a test compound that gives 50% inhibition of phosphorylated FGFRl .

Claims

1. A compound of formula formula (I):

(I) wherein

D represents CH₂;

E represents CH₂ or O;

R^N1 represents hydrogen or a Ci-C3alkyl group optionally substituted by one or more substituents selected from Ci-C₃alkoxy, cyano, hydroxyl, amino (-NH₂), mono-Ci-C3alkylamino and di-(Ci-C3alkyl)amino;

R¹ represents -F, -OH, -CN, a Ci-C3alkoxy group optionally substituted by one or more R⁸ , or -CONHR⁶;

R² represents hydrogen or a Ci-C₃alkyl group optionally substituted by one or more substituents selected from Ci-C3alkoxy, cyano, hydroxyl, amino (-NH₂), mono-Ci-C3alkylamino and di-(Ci-C3alkyl)amino;

R³ represents hydrogen, -F, or a -CONHR⁷ group; R⁴ represents hydrogen, a Ci-Cόalkyl group optionally substituted with Ci-C3alkoxy, hydroxyl, amino (-NH₂), mono-Ci-C3alkylamino and di- (Ci-C₃alkyl)amino, a C₂-C6alkenyl group optionally substituted with Ci-C3alkoxy, a C₂-Cόalkynyl group optionally substituted with Ci-C3alkoxy, a C₃-C₅cycloalkyl group optionally substituted with Ci-C₃alkoxy, a Ci-C_όalkoxy group optionally substituted with d-C₃alkoxy, hydroxyl, amino (-NH₂), mono-Ci-C3alkylamino and di-(Ci-C3alkyl)amino, -C(O)NR⁹R¹⁰, -NR¹¹R¹²,

-S(O)_yR¹³ where y is 0, 1 or 2; R⁵ represents hydrogen or a Ci-C3alkoxy group;

R⁶ represents a Ci-Cόalkyl optionally substituted by one or more R⁸; R⁷ represents hydrogen or a Ci-C₆alkyl optionally substituted by one or more R⁸; R⁹ and R¹⁰ each independently represent hydrogen, Ci-C₄alkyl or C₃-C₆cycloalkyl, or R⁹ and R¹⁰ together with the nitrogen atom to which they are attached form a 4- to 6-membered saturated heterocycle optionally comprising an additional heteratom selected from oxygen, sulphur or nitrogen wherein each R⁹ and R¹⁰ independently may be optionally substituted on carbon by one or more substituents R¹⁴ and wherein if said heterocycle contains an -NH- moiety that nitrogen may be optionally substited by a group selected from R¹⁵ ; R¹¹ and R¹² each independently represent hydrogen, d-C₄alkyl or C₃-C₆cycloalkyl, or R¹¹ and R¹² together with the nitrogen atom to which they are attached form a 4- to 6-membered saturated heterocycle optionally comprising an additional heteratom selected from oxygen, sulphur or nitrogen wherein each R¹¹ and R¹² independently may be optionally substituted on carbon by one or more substituents R¹⁶ and wherein if said heterocycle contains an -NH- moiety that nitrogen may be optionally substited by a group selected from R¹⁷ ; R¹³ represents Ci-Cβalkyl or C₃-C₆cycloalkyl; R⁸, R¹⁴ and R¹⁶ each independently is selected from halogen, Ci-C₃alkyl, Ci-C3alkoxy, Cscycloalkyl, Ci-C3alkylthio, amino (-NH₂), mono- and di- Ci-C3alkylamino, cyano, hydroxyl and trifluoromethyl; R¹⁵ and R¹⁷ each independently is selected from Ci-C_όalkyl, benzyl, Ci-C_όalkoxycarbonyl, Ci-C_όalkylcarbonyl, phenylcarbonyl, Ci-C_όalkylsulphonyl and phenylsulphonyl; and either when Q represents CR⁵, Q¹ represents N or CH; or when Q represents N, Q¹ represents CH, or a pharmaceutically acceptable salt thereof.

2. A compound or a pharmaceutically acceptable salt thereof according to Claim 1 wherein R¹ represents -F, -OH, -OMe or -CONHMe.

3. A compound or a pharmaceutically acceptable salt thereof according to Claim 1 or 2 wherein R^N1 represents hydrogen.

4. A compound or a pharmaceutically acceptable salt thereof according to any one of Claims 1 or 3 wherein R² represents hydrogen.

5. A compound or a pharmaceutically acceptable salt thereof according to any one of Claims 1 to 4 wherein R³ represents hydrogen, -F or -CONH₂.

6. A compound or a pharmaceutically acceptable salt thereof according to any one of Claims 1 to 5 wherein R⁴ is hydrogen.

7. A compound or a pharmaceutically acceptable salt thereof according to any one of Claims 1 to 6 wherein E is N or -CR⁵ wherein R⁵ is hydrogen or -OMe.

8. A compound or a pharmaceutically acceptable salt thereof according to Claim 1 wherein the compound or a pharmaceutically acceptable salt thereof is a compound of formula (Ia):

(Ia) wherein:

E represents CH₂;

R^N1 represents hydrogen;

R¹ represents -F, -OH, -OMe or a -CONHMe group;

R² represents hydrogen;

R⁴ represents hydrogen;

Q represents CR⁵ where R⁵ is hydrogen or -OMe; and either when R^3a represents hydrogen, R^3b represents hydrogen or a -CONH₂ group; or when R^3a represents -F, R^3a represents hydrogen; or a pharmaceutically acceptable salt thereof.

9. A pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt thereof, as claimed in any one of Claims 1 to 8, in association with a pharmaceutically acceptable adjuvant, diluent or carrier.

10. A process for the preparation of a pharmaceutical composition as claimed in Claim 9 which comprises mixing a compound of formula (I), or a pharmaceutically acceptable salt thereof, as defined in any one of claims 1 to 8, with a pharmaceutically acceptable adjuvant, diluent or carrier.

11. A compound of formula (I), or a pharmaceutically-acceptable salt thereof, as claimed in any one of Claims 1 to 8 for use in therapy.

12. Use of a compound of formula (I), or a pharmaceutically acceptable salt, as claimed in any one of claims 1 to 8 in the manufacture of a medicament for use in therapy.

13. A method of treating cancer which comprises administering to a patient in need thereof a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as claimed in any one of Claims 1 to 8.

14. A method of modulating FGFR activity which comprises administering to a patient in need thereof a therapeutically effective amount of a compound of formula (I), or a pharmaceutically acceptable salt thereof, as claimed in any one of Claims 1 to 8.