# Patent

## Title
2-anilino-pyrimidine derivatives as cyclin dependent kinase inhibitors

## Abstract

Compounds of the formula (I): wherein R1, R2, R3, p, q, and X are as defined within and a pharmaceutically acceptable salts and in vivo hydrolysable esters are described. Also described are processes for their preparation and their use as medicaments, particularly medicaments for producing a cell cycle inhibitory (anti-cell-proliferation) effect in a warm-blooded animal, such as man.

## Background
The invention relates to pyrimdine derivatives, or pharmaceutically acceptable salts or in vivo hydrolysable esters thereof, which possess cell-cycle inhibitory activity and are accordingly useful for their anti-cell-proliferation (such as anti-cancer) activity and are therefore useful in methods of treatment of the human or animal body. The invention also relates to processes for the manufacture of said pyrimidine derivatives to pharmaceutical compositions containing them and to their use in the manufacture of medicaments of use in the production of an anti-cell-proliferation effect in a warm-blooded animal such as man.
A family of intracellular proteins called cyclins play a central role in the cell cycle. The synthesis and degradation of cyclins is tightly controlled such that their level of expression fluctuates during the cell cycle. Cyclins bind to cyclin-dependent serine/threonine kinases (CDKs) and this association is essential for CDK (such as CDK1, CDK2, CDK4 and/or CDK6) activity within the cell. Although the precise details of how each of these factors combine to regulate CDK activity is poorly understood, the balance between the two dictates whether or not the cell will progress through the cell cycle.
The recent convergence of oncogene and tumour suppressor gene research has identified regulation of entry into the cell cycle as a key control point of mitogenesis in tumours. Moreover, CDKs appear to be downstream of a number of oncogene signalling pathways. Disregulation of CDK(activity by up regulation of cyclins and/or deletion of endogenous inhibitors appears to be an important axis between mitogenic signalling pathways and proliferation of tumour cells.
Accordingly it has been recognised that an inhibitor of cell cycle kinases, particularly inhibitors of CDK2, CDK4 and/or CDK6 (which operate at the S-phase, G1-S and G1-S phase respectively) should be of value as a selective inhibitor of cell proliferation, such as growth of mammalian cancer cells.
The present invention is based on the discovery that certain pyrimidine compounds surprisingly inhibit the effects of cell cycle kinases showing selectivity for CDK2, CDK4 and CDK6, and thus possess anti-cell-proliferation properties. Such properties are expected to be of value in the treatment of disease states associated with aberrant cell cycles and cell proliferation such as cancers (solid tumours and leukemias), fibroproliferative and differentiative disorders, psoriasis, rheumatoid arthritis, Kaposi's sarcoma, haemangioma, acute and chronic nephropathies, atheroma, atherosclerosis, arterial restenosis, autoimmune diseases, acute and chronic inflammation, bone diseases and ocular diseases with retinal vessel proliferation.
Accordingly, the present invention provides a compound of formula (I): 




wherein:

X is —O— or —S—;
R1 is selected from halo, nitro, cyano, hydroxy, amino, carboxy, carbamoyl, mercapto, C1-6alkyl, C2-6alkenyl and C2-6alkynyl;
p is 0-4; wherein the values of R1 may be the same or different;
R2 is selected from sulphamoyl and a group R4-E-;
q is 0-2; wherein the values of R2 maybe the same or different; and wherein p+q=1-5;
R3 is selected from C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C3-8cycloalkyl and heterocyclyl; wherein R3 may be optionally substituted by one or more R5; and wherein if said heterocyclyl contains an —NH— moiety that nitrogen may be optionally substituted by a group selected from R6; and wherein if said heterocyclic group contains an —NH— moiety that nitrogen may be optionally substituted by a group selected from R7;
R4 is selected from C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C3-8cycloalkyl, C3-8cycloalkylC1-6alkyl, phenyl, a heterocyclic group, phenylC1-6alkyl and (heterocyclic group)C1-6alkyl; wherein R4 may be optionally substituted on carbon by one or more R8; and wherein if said heterocyclic group contains an —NH— moiety that nitrogen may be optionally substituted by a group selected from R9;
E is selected from —C(O)—, —N(Ra)C(O)—, —C(O)N(Ra)—, —S(O)r—, —SO2N(Ra)— and —N(Ra)SO2—; wherein Ra is hydrogen or C1-6 alkyl optionally substituted by one or more R10 and r is 1-2;
R5 is selected from halo, nitro, cyano, hydroxy, trifluoromethoxy, amino, carboxy, carbamoyl, mercapto, sulphamoyl, C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C1-6alkoxy, C1-6alkanoyl, C1-6alkanoyloxy, N-(C1-6alkyl)amino, N,N-(C1-6alkyl)2amino, C1-6alkanoylamino, N-(C1-6alkyl)carbamoyl, N,N-(C1-6alkyl)2carbamoyl, C1-6alkylS(O)a wherein a is 0 to 2, C1-6alkoxycarbonyl, N-(C1-6alkyl)sulphamoyl, N,N-(C1-6alkyl)2sulphamoyl, C3-8cycloalkyl, phenyl and a heterocyclic group; wherein R5 may be optionally substituted on carbon by one or more R11; and wherein if said heterocyclic group contains an —NH— moiety that nitrogen may be optionally substituted by a group selected from R12;
R6, R7, R9 and R12 are independently selected from C1-4alkyl, C1-4alkanoyl, C1-4alkylsulphonyl, C1-4alkoxycarbonyl, carbamoyl, N-(C1-4alkyl)carbamoyl, N,N-(C1-4alkyl)carbamoyl, benzyl, benzyloxycarbonyl, benzoyl and phenylsulphonyl; wherein R6, R7, R9 and R12 may be independently optionally substituted on carbon by one or more R13;
R8 and R10 are independently selected from halo, nitro, cyano, hydroxy, trifluoromethyl, trifluoromethoxy, amino, carboxy, carbamoyl, mercapto, sulphamoyl, C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C1-6alkoxy C1-6alkanoyl, C1-6alkanoyloxy, N-(C1-6alkyl)amino, N,N-(C1-6)2amino, C1-6alkanoylamino, N-(C1-6alkyl)carbamoyl, N,N-(C1-6alkyl)2carbamoyl, C1-6alkylS(O)a wherein a is 0 to 2, C1-6alkoxycarbonyl N-(C1-6alkyl)sulphamoyl and N,N-(C1-6alkyl)2sulphamoyl; wherein R8 and R10 may be independently optionally substituted on carbon by one or more R14; and
R11, R13 and R14 are independently selected from halo, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino, carboxy, carbamoyl, mercapto, sulphamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino, ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulphinyl, ethylsulphinyl, mesyl, ethylsulphonyl, methoxycarbonyl, ethoxycarbonyl, N-methylsulphamoyl, N-ethylsulphamoyl, N,N-dimethylsulphamoyl, N,N-diethylsulphamoyl or N-methyl-N-ethylsulphamoyl;
or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.
In this specification the term “alkyl” includes both straight and branched chain alkyl groups but references to individual alkyl groups such as “propyl” are specific for the straight chain version only. For example, “C1-6alkyl” includes C1-4alkyl, C1-3alkyl, propyl, isopropyl and t-butyl. However, references, to individual alkyl groups such as ‘propyl’ are specific for the straight chained version only and references to individual branched chain alkyl groups such as ‘isopropyl’ are specific for the branched chain version only. A similar convention applies to other radicals, for example “phenylC1-6alkyl” includes phenylC1-4alkyl, benzyl, 1-phenylethyl and 2-phenylethyl. The term “halo” refers to, fluoro, chloro bromo and iodo.
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.
A “heterocyclic group” 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-oxide(s). Examples of the term “heterocyclic group” are morpholino, piperidyl, pyridyl, pyranyl, pyrrolyl, isothiazolyl, indolyl, quinolyl, thienyl, 1,3-benzodioxolyl, thiadiazolyl, piperazinyl, thiazolidinyl, pyrrolidinyl, thiomorpholino, pyrrolinyl, homopiperazinyl, 3,5-dioxapiperidinyl, tetraliydropyranyl, imidazolyl, pyrimidyl, pyrazinyl, pyridazinyl, isoxazolyl, N-methylpyrrolyl, 4-pyridone, 1-isoquinblone; 2-pyrrolidone, 4-thiazolidone, pyridine-N-oxide and quinoline-N-oxide. In one aspect of the invention a “heterocyclic group” is a saturated, partially saturated or unsaturated, mono or bicyclic ring containing 5 or 6 atoms of which at least one atom is chosen from nitrogen, sulphur or oxygen, 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 maybe optionally oxidised to form (the) S-oxide(s). In another aspect of the invention a “heterocyclic group” is morpholinyl, tetrahydrofuryl, piperidinyl, pyridyl, imidazolyl, piperazinyl, pyrrolidinyl, triazolyl, dioxanyl and dioxolanyl. In a further aspect of the invention, a “heterocyclic group” is morpholinyl, piperidinyl and tetrahydrofuryl.
A “heterocyclyl” is a saturated 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-oxide(s). Examples of the term “heterocyclyl” are morpholinyl, piperidyl, piperazinyl, thiazolidinyl, pyrrolidinyl, thiomorpholinyl, 1,1-dioxothiomorpholinyl, homopiperazinyl, tetrahydropyranyl, 2-pyrrolidone and 4-thiazolidone. In one aspect of the invention a “heterocyclyl” is a saturated monocyclic ring containing 5 or 6 atoms of which at least one atom is chosen from nitrogen, sulphur or oxygen, 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-oxide(s). In another aspect of the invention a “heterocyclyl” is tetrahydrofuryl.
An example of “C1-6alkanoyloxy” is acetoxy. Examples of “C1-6alkoxycarbonyl” include C1-4alkoxycarbonyl, methoxycarbonyl, ethoxycarbonyl, n- and t-butoxycarbonyl. Examples of “C1-6alkoxy” include methoxy, ethoxy and propoxy. Examples of “C1-6alkanoylamino” include formamido, acetamido and propionylamino. Examples of “C1-6alkylS(O)a wherein a is 0 to 2” include C1-4alkylsulphonyl, methylthio, ethylthio, methylsulphinyl, ethylsulphinyl, mesyl and ethylsulphonyl. Examples of “C1-6alkylS(O)r wherein r is 1 to 2” include methylsulphinyl, ethylsulphinyl, mesyl and ethylsulphonyl. Examples of “C1-6alkanoyl” include C1-4alkanoyl, propionyl and acetyl. Examples of “N—C1-6alkylamino” include methylamino and ethylamino. Examples of “N,N-(C1-6alkyl)2amino” include di-N-methylamino, di-(N-ethyl)amino and N-ethyl-N-methylamino. Examples of “C2-6alkenyl” are vinyl, allyl and 1-propenyl. Examples of “C2-6alkynyl” are ethynyl, 1-propynyl and 2-propynyl. Examples of “N-(C1-6alkyl)sulphamoyl” are N-(methyl)sulphamoyl and N-(ethyl)sulphamoyl. Examples of “N-(C1-6alkyl)2sulphamoyl” are N,N-(dimethyl)sulphamoyl and N-(methyl)-N-(ethyl)sulphamoyl. Examples of “N-(C1-6alkyl)carbamoyl” are N-(C1-4alkyl)carbamoyl, methylaminocarbonyl and ethylaminocarbonyl. Examples of “N,N-(C1-6alkyl)2carbamoyl” are N,N-(C1-4alkyl)2carbamoyl, dimethylaminocarbonyl and methylethylaminocarbonyl. Examples of “C3-8cycloalkyl” are cyclopropyl, cyclobutyl, cyclopropyl and cyclohexyl. Examples of “(heterocyclic group)C1-6alkyl” include pyridylmethyl, 3-morpholinopropyl and 2-pyrimid-2-ylethyl. Examples of “C3-8cycloalkylC1-6alkyl” are cyclopropylethyl, cyclobutylmethyl, 2-cyclopropylpropyl and cyclohexylethyl.
A suitable pharmaceutically acceptable salt of a compound of the invention is, for example, an acid-addition salt of a compound of the invention which is sufficiently basic, for example, an acid-addition salt with, for example, an inorganic or organic acid, for example hydrochloric, hydrobromic, sulphuric, phosphoric, trifluoroacetic, citric or maleic acid. In addition a suitable pharmaceutically acceptable salt of a compound of the invention which is sufficiently acidic is an alkali metal salt, for example a sodium or potassium salt, an alkaline earth metal salt, for example a calcium or magnesium salt, an ammonium salt or a salt with an organic base which affords a physiologically-acceptable cation, for example a salt with methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris-(2-hydroxyethyl)amine.
The compounds of the formula (I) may be administered in the form of an in vivo hydrolysable ester which is broken down in the human or animal body to give a compound of the formula (I).
An in vivo hydrolysable ester of a compound of the formula (I) containing carboxy or hydroxy group is, for example, a pharmaceutically acceptable ester which is hydrolysed in the human or animal body to produce the parent acid or alcohol. Suitable pharmaceutically acceptable esters for carboxy include C1-6alkoxymethyl esters for example methoxymethyl, C1-6alkanoyloxymethyl esters for example pivaloyloxymethyl, phthalidyl esters, C3-8cycloalkcoxycarbonyloxyC1-6alkyl esters for example 1-cyclohexylcarbonyloxyethyl; 1,3-dioxolen-2-onylmethyl esters for example 5-methyl-1,3-dioxolen-2-onylmethyl; and C1-6alkoxycarbonyloxyethyl esters for example 1-methoxycarbonyloxyethyl and may be formed at any carboxy group in the compounds of this invention.
An in vivo hydrolysable ester of a compound of the formula (I) containing a hydroxy group includes inorganic esters such as phosphate esters and α-acyloxyalkyl ethers and related compounds which as a result of the in vivo hydrolysis of the ester breakdown to give the parent hydroxy group. Examples of α-acyloxyalkyl ethers include acetoxymethoxy and 2,2-dimethylpropionyloxy-methoxy. A selection of in vivo hydrolysable ester forming groups for hydroxy include alkanoyl, benzoyl phenylacetyl and substituted benzoyl and phenylacetyl, alkoxycarbonyl (to give alkyl carbonate esters), dialkylcarbamoyl and N-(dialkylaminoethyl)-N-alkylcarbamoyl (to give carbamates), dialkylaminoacetyl and carboxyacetyl. Examples of substituents on benzoyl include morpholino and piperazin-1-yl linked from a ring nitrogen atom via a methylene group to the 3- or 4- position of the benzoyl ring.
Some compounds of the formula (I) may have chiral centres and/or geometric isomeric centres (E- and Z-isomers), and it is to be understood that the invention encompasses all such optical, diastereoisomers and geometric isomers that possess CDK inhibitory activity.
The invention relates to any and all tautomeric forms of the compounds of the formula (I) that possess CDK inhibitory activity.
It is also to be understood that certain compounds of the formula (I) can exist in solvated as well as unsolvated forms such as, for example, hydrated forms. It is to be understood that the invention encompasses all such solvated forms which possess CDK inhibitory activity.
Further values of R1, R2, R3, p and q are as follows. Such values may be used where appropriate with any of the definitions, claims or embodiments defined hereinbefore or hereinafter.
X is —O—.
X is —S—.
R1 is halo or C1-2alkyl.
R1 is fluoro, or chloro or methyl.
R1 is fluoro or chloro.
p is 0-2; wherein the values of R1 may be the same or different.
p is 0 or 1.
p is 0.
p is 1.
p is 2; wherein the values of R1 may be the same or different.
p is 1 and R1 is meta to the amino group in the aniline of formula (I).
R2 is sulphamoyl or a group R4-E-; wherein:
R4 is selected from C1-6alkyl, C2-6alkenyl, C3-8cycloalkyl and (heterocyclic group)C1-6alkyl; wherein R4 may be optionally substituted on carbon by one or more R8;
E is selected from —S(O)r— and —N(Ra)SO2—; wherein Ra is hydrogen or C1-6alkyl and r is 2; and
R8 is selected from halo, C1-6alkoxy and N-(C1-6alkyl)amino.
R2 is sulphamoyl or a group R4-E-; wherein:
R4 is selected from ethyl, propyl, butyl, allyl, cyclopropyl, tetrahydrofur-2-ylmethyl and piperidinoethyl; wherein R4 may be optionally substituted on carbon by one or more R8;
E is selected from —S(O)2—, —NHSO2— and —N(e)SO2—; and
R8 is selected from fluoro, methoxy and isopropylamino.
R2 is sulphamoyl, N,N-dimethylsulphamoyl, N-(cyclopropyl)sulphamoyl, N-(2-methoxyethyl)sulphamoyl, N-(3-methoxypropyl)sulphamoyl, N-(2,2,2-trifluoroethyl)sulphamoyl, ethylsulphonyl, 4-(isopropylamino)butylsulphonyl, N-(allyl)sulphamoyl N-(tetrahydrofur-2-ylmethyl)sulphamoyl and N-(2-piperidinoethyl)sulphamoyl.
R2 is sulphamoyl or a group R4-E-; wherein:
R4 is selected from C1-6alkyl, C2-6alkenyl, C3-8 cycloalkyl, C3-8cycloalkylC1-6alkyl, and (heterocyclic group)C1-6alkyl; wherein R4 may be optionally substituted on carbon by one or more R8;
E is selected from —S(O)r— and —N(Ra)SO2—; wherein Ra is hydrogen or C1-6alkyl and r is 2; and
R8 is selected from halo, C1-6alkoxy and N-(C1-6alkyl)amino.
R2 is sulphamoyl or a group R4-E-; wherein:
R4 is selected from methyl, ethyl, propyl, butyl, allyl, cyclopropyl, cyclobutyl, cyclopropylmethyl, tetrahydrofur-2-ylmethyl and piperidinoethyl; wherein R4 may be optionally substituted on carbon by one or more R8;
E is selected from —S(O)2—, —NHSO2— and —N(Me)SO2—; and
R8 is selected from fluoro, methoxy and isopropylamino.
R2 is sulphamoyl, N,N-dimethylsulphamoyl, N-(cyclopropyl)sulphamoyl, N-(2-methoxyethyl)sulphamoyl, N-(3-methoxypropyl)sulphamoyl, N-(2,2,2-trifluoroethyl)sulphamoyl, ethylsulphonyl, 4-(isopropylamino)butylsulphonyl, N-(allyl)sulphamoyl N-(tetrahydrofur-2-ylmethyl)sulphamoyl, N-(2-piperidinoethyl)sulphamoyl, N-(methyl)sulphamoyl, N-(methyl)-N-(2-methoxyethyl)sulphamoyl, N-(cyclobutyl)sulphamoyl and N-(cyclopropylmethyl)sulphamoyl.
E is —NHSO2—, —N(Me)SO2— or —SO2—.
E is —NHSO2— or —SO2—.
E is —NHSO2—.
E is —SO2—.
q is 0 or 1.
q is 0.
q is 1.
q is 2; wherein the values of R2 may be the same or different.
p+q=1 or 2.
p+q=1.
q is 1 and R2 is meta or para to the amino group in the aniline of formula (I).
q is 1 and R2 is para to the amino group in the aniline of formula (I).
R3 is selected from C1-6alkyl, C2-6alkynyl and heterocyclyl; wherein R3 may be optionally substituted by one or more R5; wherein:
R5 is selected from C1-6alkoxy, C3-8cycloalkyl and a heterocyclic group; wherein R5 may be optionally substituted on carbon by one or more R11; and
R11 is methoxy.
R3 is selected from methyl, ethyl, isopropyl, butyl, 2-propynyl, 2-butynyl, 3-butynyl, 3-butyn-2-yl and tetrahydrofuryl; wherein R3 may be optionally substituted by one or more R5; wherein:
R5 is selected from methoxy, ethoxy, cyclopropyl, morpholinyl and tetrahydrofuryl; wherein R5 maybe optionally substituted on carbon by one or more R11; and
R11 is methoxy.
R3 is selected from methoxy, ethoxy, isopropoxy, butoxy, butylthio, 2-(2-methoxyethoxy)ethyl, 2-methoxyethoxy, tetrahydrofur-3-yloxy, tetrahydrofur-3-ylmethoxy, cyclopropylmethoxy, 2-propynyloxy, 2-butynyloxy, 3-butynyloxy, 3-butyn-2-yloxy and 2-morpholinoethoxy.
R3—X— is selected from methoxy, ethoxy, isopropoxy, butoxy, butylthio, 2-(2-methoxyethoxy)ethoxy, 2-methoxyethoxy, tetrahydrofur-3-yloxy, tetrahydrofur-3-ylmethoxy, cyclopropylmethoxy, 2-propynyloxy, 2-butynyloxy, 3-butynyloxy, 3-butyn-2-yloxy and 2-morpholinoethoxy.
R3 is selected from methyl, ethyl, isopropyl, butyl, 2-(2-methoxyethoxy)ethyl, 2methoxyethyl, tetrahydrofur-3-ylyl, tetrahydrofur-3-ylmethyl, cyclopropylmethyl, 2-propynyl, 2-butynyl, 3-butynyl, 3-butyn-2-yl and 2-morpholinoethyl.
R3 is selected from methyl, ethyl, isopropyl, butyl, but-2-yl, 2-propynyl, 2-butynyl, 3-butynyl, 3-butyn-2-yl and tetrahydrofuryl; wherein R3 may be optionally substituted by one or more R5; wherein:
R5 is selected from methoxy, ethoxy, cyclopropyl, morpholinyl and tetrahydrofuryl; wherein R5 maybe optionally substituted on carbon by one or more R11; and
R11 is methoxy.
R3 is selected from methyl, ethyl, isopropyl, butyl, but-2-yl, 2-(2-methoxyethoxy)ethyl, 2-methoxyethyl, tetrahydrofur-3-ylyl, tetrahydrofur-3-ylmethyl, cyclopropylmethyl, 2-propynyl, 2-butynyl, 3-butynyl, 3-butyn-2-yl and 2-morpholinoethyl.
R3—X— is selected from methoxy, ethoxy, isopropoxy, butoxy, but-2-yloxy, butylthio, 2-(2-methoxyethoxy)ethoxy, 2-methoxyethoxy, tetrahydrofur-3-yloxy, tetrahydrofur-3-ylmethoxy, cyclopropylmethoxy, 2-propynyloxy, 2-butynyloxy, 3-butynyloxy, 3-butyn-2-yloxy and 2-morpholinoethoxy.
Therefore in one aspect of the invention, there is provided a compound of formula (I) as depicted above wherein:
X is —O— or —S—;
p is 0;
R2 is sulphamoyl or a group R4-E-; wherein:
R4 is selected from C1-6alkyl, C2-6alkenyl, C3-8cycloalkyl and (heterocyclic group)C1-6alkyl; wherein R4 may be optionally substituted on carbon by one or more R8;
E is selected from —S(O)r— and —N(Ra)SO2—; wherein Ra is hydrogen or C1-6alkyl and r is 2;
R8 is selected from halo, C1-6alkoxy and N-(C1-6alkyl)amino;
q is 1;
R3 is selected from C1-6alkyl, C2-6alkynyl and heterocyclyl; wherein R3 may be optionally substituted by one or more R5; wherein:
R5 is selected from C1-6alkoxy, C3-8 cycloalkyl and a heterocyclic group; wherein R5 may be optionally substituted on carbon by one or more R11; and
R11 is methoxy;
or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.
Therefore in a further aspect of the invention, there is provided a compound of formula (I) as depicted above wherein:
X is —O— or —S—;
p is 0;
R2 is sulphamoyl or a group R4-E-; wherein:
R4 is selected from ethyl, propyl, butyl, allyl, cyclopropyl, tetrahydrofur-2-ylmethyl and piperidinoethyl; wherein R4 may be optionally substituted on carbon by one or more R8;
E is selected from —S(O)2—, —NHSO2 and —N(Me)SO2—;
R8 is selected from fluoro, methoxy and isopropylamino;
q is 1;
R3 is selected from methyl, ethyl, isopropyl, butyl, 2-propynyl, 2-butynyl, 3-butynyl, 3-butyn-2-yl and tetrahydrofuryl; wherein R3 may be optionally substituted by one or more R5; wherein:
R5 is selected from methoxy, ethoxy, cyclopropyl, morpholinyl and tetrahydrofuryl; wherein R5 may be optionally substituted on carbon by one or more R11; and
R11 is methoxy;
or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.
Therefore in another further aspect of the invention, there is provided a compound of formula (I) (as depicted above); wherein:
X is —O— or —S—;
p is 0;
R2 is sulphamoyl, N,N-dimethylsulphamoyl, N-(cyclopropyl)sulphamoyl, N-(2-methoxyethyl)sulphamoyl, N-(3-methoxypropyl)sulphamoyl, N-(2,2,2-trifluoroethyl)sulphamoyl, ethylsulphonyl, 4-(isopropylamino)butylsulphonyl, N-(allyl)sulphamoyl, N-(tetrahydrofur-2-ylmethyl)sulphamoyl and N-(2-piperidinoethyl)sulphamoyl;
q is 1; and
R3 is selected from methoxy, ethoxy, isopropoxy, butoxy, butylthio, 2-(2-methoxyethoxy)ethyl, 2-methoxyethoxy, tetrahydrofur-3-yloxy, tetrahydrofur-3-ylmethoxy, cyclopropylmethoxy, 2-propynyloxy, 2-butynyloxy, 3-butynyloxy, 3-butyn-2-yloxy and 2-morpholinoethoxy;
or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.
Therefore in one aspect of the invention, there is provided a compound of formula (I) as depicted above wherein:
X is —O— or —S—;
p is 0;
R2 is sulphamoyl or a group R4-E-; wherein:
R4 is selected from C1-6alkyl, C2-6alkenyl, C3-8cycloalkyl, C3-8cycloalkylC1-6alkyl, and (heterocyclic group)C1-6alkyl; wherein R4 may be optionally substituted on carbon by one or more R8;
E is selected from —S(O)r— and —N(Ra)SO2—; wherein Ra is hydrogen or C1-6alkyl and r is 2;
R8 is selected from halo, C1-6alkoxy and N-(C1-6alkyl)amino;
q is 1;
R3 is selected from C1-6alkyl, C2-6alkynyl and heterocyclyl; wherein R3 may be optionally substituted by one or more R5; wherein:
R5 is selected from C1-6alkoxy, C3-8cycloalkyl and a heterocyclic group; wherein R5 may be optionally substituted on carbon by one or more R11; and
R11 is methoxy;
or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.
Therefore in a further aspect of the invention, there is provided a compound of formula (I) as depicted above wherein:
X is —O— or —S—;
p is 0;
R2 is sulphamoyl or a group R4-E-; wherein:
R4 is selected from methyl, ethyl, propyl, butyl, allyl, cyclopropyl, cyclobutyl, cyclopropylmethyl, tetrahydrofur-2-ylmethyl and piperidinoethyl; wherein R4 may be optionally substituted on carbon by one or more R8;
E is selected from —S(O)2—, —NHSO2— and —N(Me)SO2—;
R8 is selected from fluoro, methoxy and isopropylamino;
q is 1;
R3 is selected from methyl, ethyl, isopropyl, butyl, but-2-yl, 2-propynyl, 2-butynyl, 3-butynyl, 3-butyn-2-yl and tetrahydrofuryl; wherein R3 may be optionally substituted by one or more R5; wherein:
R5 is selected from methoxy, ethoxy, cyclopropyl, morpholinyl and tetrahydrofuryl; wherein R5 may be optionally substituted on carbon by one or more R11, and
R11 is methoxy;
or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.
Therefore in another further aspect of the invention, there is provided a compound of formula (I) (as depicted above); wherein:
X is —O— or —S—;
p is 0;
R2 is sulphamoyl, N,N-dimethylsulphamoyl, N-(cyclopropyl)sulphamoyl, N-(2-methoxyethyl)sulphamoyl, N-(3-methoxypropyl)sulphamoyl, N-(2,2,2-trifluoroethyl)sulphamoyl, ethylsulphonyl, 4-(isopropylamino)butylsulphonyl, N-(allyl)sulphamoyl N-(tetrahydrofur-2-ylmethyl)sulphamoyl, N-(2-piperidinoethyl)sulphamoyl, N-(methyl)sulphamoyl, N-(methyl)-N-(2-methoxyethyl)sulphamoyl, N-(cyclobutyl)sulphamoyl and N-(cyclopropylmethyl)sulphamoyl;
q is 1; and
R3 is selected from methyl, ethyl, isopropyl, butyl, but-2-yl, 2-(2-methoxyethoxy)ethyl, 2-methoxyethyl, tetrahydrofur-3-ylyl, tetrahydrofur-3-ylmethyl, cyclopropylmethyl, 2-propynyl, 2-butynyl, 3-butynyl, 3-butyn-2-yl and 2-morpholinoethyl;
or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.
In another aspect of the invention, preferred, compounds of the invention are any one of the Examples or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.
Preferred aspects of the invention are those which relate to the compound of formula (I) or a pharmaceutically acceptable salt thereof.
Another aspect of the present invention provides a process for preparing a compound of formula (I) or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof which process (wherein R1, R2, R3, p and q are, unless otherwise specified, as defined in formula (I)) comprises of:

    a) reaction of a pyrimidine of formula (II): 




wherein L is a displaceable group; with an aniline of formula (III): 




    b) reacting a pyrimidine of formula (IV): 




wherein L is a displaceable group; with an compound of formula (V):

 R3XH  (V)

or

    c) for compounds of formula (I) where R2 is sulphamoyl or a group R4-E- and E is —NHSO2—; reacting a pyrimidine of formula (VI): 




wherein Z is a displaceable group; with an amine of formula (VII):

R4—NH2  (VII)

    d) by converting a compound of formula (VIII): 




wherein Rb is hydrogen or t-butyl; into a compound of formula (I); and thereafter if necessary:

    i) converting a compound of the formula (I) into another compound of the formula (I);
    ii) removing any protecting groups;
    iii) forming a pharmaceutically acceptable salt or in vivo hydrolysable ester.


L is a displaceable group, suitable values for L are for example, a halogeno, an optionally substituted phenoxy or sulphonyloxy group, for example a chloro, bromo, phenoxy, 4-chlorophenoxy, methanesulphonyloxy or toluene-4-sulphonyloxy group.
Z is a displaceable group, suitable values for Z are for example, a halogeno group, for example a fluoro, chloro or bromo group. Preferably Z is fluoro.
Preferably T is S.
Specific reaction conditions for the above reactions are as follows.
a) Pyrimidines of formula (I) and anilines of formula (III) may be reacted together:


    i) in the presence of a suitable solvent for example a ketone such as acetone or an alcohol such as ethanol or butanol or an aromatic hydrocarbon such as toluene or N-methyl pyrrolidine, optionally in the presence of a suitable acid 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) and at a temperature in the range of 0° C. to reflux, preferably reflux; or
    ii) under standard Buchwald conditions (for example see J. Am. Chem. Soc., 118, 7215; J. Am. Chem. Soc., 119, 8451; J. Org. Chem., 62, 1568 and 6066) for example in the presence of palladium acetate, in a suitable solvent for example an aromatic solvent such as toluene, benzene or xylene with a suitable base for example an inorganic base such as caesium carbonate or an organic base such as potassium-t-butoxide, in the presence of a suitable ligand such as 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl and at a temperature in the range of 25 to 80° C.


Pyrimidines of the formula (II) and anilines of formula (III) are commercially available compounds, or they are known in the literature, or they are prepared by standard processes known in the art.
b) Pyrimidines of formula (IV) and compounds of formula (V) may be reacted together:


    i) under the conditions described under process a) i); or
    ii) in the presence of a suitable solvent for example a ketone such as acetone or an alcohol such as ethanol or butanol or an aromatic hydrocarbon such as toluene or N-methyl pyrrolidine, in the presence of a suitable base for example an alkoxide such as sodium ethoxide or potassium t-butoxide, and at a temperature in the range of 0° C. to reflux, preferably reflux.


Pyrimidines of the formula (IV) and compounds of formula (V) are commercially available compounds, or they are known in the literature, or they are prepared by standard processes known in the art.
c) Compounds of formula (VI) and compounds of formula (VII) may be reacted together in the presence of a base for example an inorganic base such as caesium carbonate in the presence of an inert solvent such as toluene or tetrahydrofuran, or in the presence of an organic base such as excess (VI) and at a temperature in the range of 25 to 80° C.
Compounds of formula (VI) wherein Z is fluoro may be prepared according to the following scheme: 




Compounds of formula (VIa) and (VII) are commercially available compounds, or they are known in the literature, or they are prepared by standard processes known in the art.
d) Compounds of formula (VII may be converted into compounds of formula (I) under standard conditions, for example in the presence, of trifluoroacetic anhydride (where Rb is hydrogen) or thionyl chloride at a temperature in the range of 25 to 100° C.
Compounds of formula (VIII may be prepared according to the following scheme: 




Compounds of formula (VIIIa) and (VIIIc) are commercially available compounds, or they are known in the literature, or they are prepared by standard processes known in the art.
It will be appreciated that certain of the various ring substituents in the compounds of the present invention may be introduced by standard aromatic substitution reactions or generated by conventional functional group modifications either prior to or immediately following the processes mentioned above, and as such are included in the process aspect of the invention. Such reactions and modifications include, for example, introduction of a substituent by means of an aromatic substitution reaction, reduction of substituents, 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 aluminum 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 modifications include the reduction of a nitro group to an amino group by for example, catalytic hydrogenation with a nickel catalyst or treatment with iron in the presence of hydrochloric acid with heating; oxidation of alkylthio to alkylsulphinyl or alkylsulphonyl.
It will also be appreciated that in some of the reactions mentioned herein it may be necessary/desirable to protect any sensitive groups in the compounds. The instances where protection is necessary or desirable and suitable methods for protection are known to those skilled in the art. Conventional protecting groups may be used in accordance with standard practice (for illustration see T. W. Green, Protective Groups in Organic Synthesis, John Wiley and Sons, 1991). Thus, if reactants include groups such as amino, carboxy or hydroxy it may be desirable to protect the group in some of the reactions mentioned herein.
A suitable protecting group for an amino or alkylamino group is, for example, an acyl group, for example an alkanoyl group such as acetyl, an alkoxycarbonyl group, for example a methoxycarbonyl, ethoxycarbonyl or t-butoxycarbonyl group, an arylmethoxycarbonyl group, for example benzyloxycarbonyl, or an aroyl group, for example benzoyl. The deprotection conditions for the above protecting groups necessarily vary with the choice of protecting group. Thus, for example, an acyl group such as an alkanoyl or alkoxycarbonyl group or an aroyl group maybe removed for example, by hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium or sodium hydroxide. Alternatively an acyl group such as a t-butoxycarbonyl group may be removed, for example, by treatment with a suitable acid as hydrochloric, sulphuric or phosphoric acid or trifluoroacetic acid and an arylmethoxycarbonyl group such as a benzyloxycarbonyl group may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon, or by treatment with a Lewis acid for example boron tris(trifluoroacetate). A suitable alternative protecting group for a primary amino group is, for example, a phthaloyl group which may be removed by treatment with an alkylamine, for example dimethylaminopropylamine, or with hydrazine.
A suitable protecting group for a hydroxy group is, for example, an acyl group, for example an alkanoyl group such as acetyl, an aroyl group, for example benzoyl, or an arylmethyl group, for example benzyl. The deprotection conditions for the above protecting groups will necessarily vary with the choice of protecting group. Thus, for example, an acyl group such as alkanoyl or an aroyl group may be removed, for example, by hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium or sodium hydroxide. Alternatively an arylmethyl group such as a benzyl group may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon.
A suitable protecting group for a carboxy group is, for example, an esterifying group, for example a methyl or an ethyl group which may be removed, for example, by hydrolysis with a base such as sodium hydroxide, or for example a t-butyl group which maybe removed, for example, by treatment with an acid, for example an organic acid such as trifluoroacetic acid, or for example a benzyl group which may be removed, for example, by hydrogenation over a catalyst such as palladium-on-carbon.
The protecting groups may be removed at any convenient stage in the synthesis using conventional techniques well known in the chemical art.
As stated hereinbefore the compounds defined in the present invention possesses anti-cell-proliferation activity such as anti-cancer activity which is believed to arise from the CDK inhibitory activity of the compound. These properties may be assessed, for example, using the assay as set out in WO 01/14375, page 27-30, as follows:
Assay
The following abbreviations have been used:

    HERPES is N-[2-Hydroxyethyl]piperazine-N′-[2-ethanesulfonic acid]
    DTT is Dithiothretiol
    PMSF is Phenylmethylsulfonyl fluoride


The compounds were tested in an in vitro kinase assay in 96 well format using Scintillation Proximity Assay (SPA—obtained form Amersham) for measuring incorporation of [γ-33-P]-Adenosine Triphosphate into a test substrate (GST-Retinoblastoma protein; GST-Rb). In each well was placed the compound to be tested (diluted in DMSO and water to correct concentrations) and in control wells either roscovitine as an inhibitor control or DMSO as a positive control.
Approximately 0.2 μl of CDK2/Cyclin E partially-purified enzyme (amount dependent on enzyme activity) diluted in 25 μl incubation buffer was added to each well then 20 μl of GST-Rb/ATP/ATP33 mixture (containing 0.5 μg GST-Rb and 0.2 μM ATP and 0.14 μCi [γ-33-P]-Adenosine Triphosphate in incubation buffer), and the resulting mixture shaken gently, then incubated at room temperature for 60 minutes.
To each well was then added 150 μL stop solution containing (0.8 mg/well of Protein A-PVT SPA bead (Amersham)), 20 pM/well of Anti-Glutathione Transferase, Rabbit IgG (obtained from Molecular Probes), 61 mM EDTA and 50 mM HEPES pH 7.5 containing 0.05% sodium azide.
The plates were sealed with Topseal-S plate sealers, left for two hours then spun at 2500 rpm, 1124xg., for 5 minutes. The plates were read on a Topcount for 30 seconds per well.
The incubation buffer used to dilute the enzyme an substrate mixes contained 50 mM HEPES pH7.5, 10 mM MnCl2, 1 mM DTT, 100 μM Sodium vanadate, 100 μM NaF, 10 mM Sodium Glycerophosphate, BSA (1 mg/ml final).
Test Substrate
In this assay only part of the retinoblastoma protein (Science 1987 Mar13;235(4794):1394-1399; Lee W. H., Bookstein R., Hong F., Young L. J., Shew J. Y., Lee E. Y.) was used, fused to a GST tag. PCR of retinoblastoma gene encoding amino acids 379-928 (obtained from retinoblastoma plasmid ATCC pLRbRNL) was performed, and the sequence cloned into pGEX 2T fusion vector (Smith D. B. and Johnson, K. S. Gene 67, 31 (1988); which contained a tac promoter for inducible expression, internal lac Iq gene for use in any E.Coli host, and a coding region for thrombin cleavage—obtained from Pharmacia Biotech) which was used to amplify amino acids 792-928. This sequence was again cloned into pGEX 2T.
The retinoblastoma 792-928 sequence so obtained was expressed in E.Coli (BL21 (DE3) pLysS cells) using standard inducible expression techniques, and purified as follows.
E.coli paste was resuspended in 10 ml/g of NETN buffer (50 mM Tris pH 7.5, 120 mM NaCl, 1 mM EDTA, 0.5% v/v NP-40, 1 mM PMSF, 1 ug/ml leupeptin, 1 ug/ml aprotinin and 1 ug/ml pepstatin) and sonicated for 2×45 seconds per 100 ml homogenate. After centrifugation, the supernatant was loaded onto a 10 ml glutathione Sepharose column (Pharmacia Biotech, Herts, UK), and washed with NETN buffer. After washing with kinase buffer (50 mM HEPES pH 7.5, 10 mM MgC12, 1 mM DTT, 1 mM PMSF, 1 ug/ml leupeptin, 1 ug/ml aprotinin and 1 ug/ml pepstatin) the protein was eluted with 50 mM reduced glutathione in kinase buffer. Fractions containing GST-Rb(792-927) were pooled and dialysed overnight against kinase buffer. The final product was analysed by Sodium Dodeca Sulfate (SDS) PAGE (polyacrylamide gel) using 8-16% Tris-Glycine gels (Novex, San Diego, USA).
CDK2 and Cyclin E
The open reading frames of CDK2 and Cyclin E were isolated by reverse transcriptase-PCR using HeLa cell and activated T cell mRNA as a template and cloned into the insect expression vector pVL1393 (obtained from Invitrogen 1995 catalogue number: V1392-20). CDK2 and cyclin E were then dually expressed [using a standard virus Baculogold co-infection technique] in the insect SF21 cell system (Spondoptera Frugiperda cells derived from ovarian tissue of the Fall Army Worm—commerically available).
Example Production of Cyclin E/CDK2
The following Example provides details of the production of Cyclin E/CDK2 in SF21 cells (in TC100+10% FBS(TCS)+0.2% Pluronic) having dual infection MOI 3 for each virus of Cyclin E & CDK2.
SF21 cells grown in a roller bottle culture to 2.33×106 cells/ml were used to inoculate 10×500 ml roller bottles at 0.2×10E6 cells/ml. The roller bottles were incubated on a roller rig at 28° C.
After 3 days (72 hrs.) the cells were counted, and the average from 2 bottles found to be 1.86×10E6 cells/ml. (99% viable). The cultures were then infected with the dual viruses at an MOI 3 for each virus.
The viruses were mixed together before addition to the cultures, and the cultures returned to the roller rig 28° C.
After 2 days (48 hrs.) post infection the 5 Litres of culture was harvested. The total cell count at harvest was 1.58×10E6 cells/ml.(99% viable). The cells were spun out at 2500 rpm, 30 mins., 4° C. in Heraeus Omnifuge 2.0 RS in 250 ml. lots. The supernatant was discarded.
Partial Co-purification of Cdk2 and Cyclin E
Sf21 cells were resuspended in lysis buffer (50 mM Tris pH 8.2, 10 mM MgCl2, 1 mM DTT, 10 mM glycerophosphate, 0.1 mM sodium orthovanadate, 0.1 mM NaF, 1 mM PMSF, 1 ug/ml leupeptin and 1 ug/ml aprotinin) homogenised for 2 minutes in a 10 ml Dounce homgeniser. After centrifugation, the supernatant was loaded onto a Poros HQ/M 1.4/100 anion exchange column (PE Biosystems, Hertford, UK). Cdk2 and Cyclin E were coeluted at the beginning of a 0-1 M NaCl gradient (run in lysis buffer minus protease inhibitors) over 20 column volumes. Co-elution was checked by western blot using both anti-Cdk2 and anti-Cyclin E antibodies (Santa Cruz Biotechnology, California, US).
By analogy, assays designed to assess inhibition of CDK4 and CDK6 may be constructed. CDK2 (EMBL Accession No. X62071) may be used together with Cyclin A or Cyclin E (see EMBL Accession No. M73812), and further details for such assays are contained in PCT International Publication No. WO99/21845, the relevant Biochemical & Biological Evaluation sections of which are hereby incorporated by reference.
Although the pharmacological properties of the compounds of the formula (I) vary with structural change, in general activity possessed by compounds of th formula (I) in this assay may be demonstrated at IC50 concentrations or doses in the range 250 μM to 1 nM.
The in vivo activity of the compounds of formula (I) may be measured in the SRB assay as set out in WO 01/14375, page 30-31, as follows:
The in vivo activity of the compounds of the present invention may be assessed by standard techniques, for example by measuring inhibition of cell growth and assessing cytotoxicity.
Inhibition of cell growth may be measured by staining cells with Sulforhodamine B (SRB), a fluorescent dye that stains proteins and therefore gives an estimation of amount of protein (i.e. cells) in a well (see Boyd, M. R. (1989) Status of the NCI preclinical antitumour drug discovery screen. Prin. Prac Oncol 10.1-12). Thus, the following details are provided of measuring inhibition of cell growth:
cells were plated in appropriate medium in a volume of 100 ml in 96 well plates; media was Dulbecco's Modified Eagle media for MCF-7, UT-1B and SK-UT-1. The cells were allowed to attach overnight, then inhibitor compounds were added at various concentrations in a maximum concentration of 1% DMSO (v/v). A control plate was assayed to give a value for cells before dosing. Cells were incubated at 37° C., (5% CO2) for three days.
At the end of three days TCA was added to the plates to a final concentration of 16% (v/v). Plates were then incubated at 4° C. for 1 hour, the supernatant removed and the plates washed in tap water. After drying, 100 ml SRB dye (0.4% SRB in 1% acetic acid) was added for 30 minutes at 37° C. Excess SRB was removed and the plates washed in 1% acetic acid. The SRB bound to protein was solubilised in 10 mM Tris pH7.5 and shaken for 30 minutes at room temperature. The ODs were read at 540 nm, and the concentration of inhibitor causing 50% inhibition of growth was determined from a semi-log plot of inhibitor concentration versus absorbance. The concentration of compound that reduced the optical density to below that obtained when the cells were plated at the start of the experiment gave the value for toxicity.
Typical IC50 values for compounds of the invention when tested in the SRB assay are in the range 1 mM to 1 nM.
According to a further aspect of the invention there is provided a pharmaceutical composition which comprises a pyrimidine derivative of the formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, as defined hereinbefore in association with a pharmaceutically-acceptable diluent or carrier.
The composition may be in a form suitable for oral administration, for example as a tablet or capsule, for parenteral injection (including intravenous, subcutaneous, intramuscular, intravascular or infusion), as a sterile solution, suspension or emulsion, for topical administration as an ointment or cream or for rectal administration as a suppository.
In general the above compositions may be prepared in a conventional manner using conventional excipients.
The compound of formula (I) will normally be administered to a warm-blooded animal at a unit dose within the range 5-5000 mg per square meter body area of the animal, i.e. approximately 0.1-100 mg/kg, and this normally provides a therapeutically-effective dose. A unit dose form such as a tablet or capsule will usually contain, for example 1-250 mg of active ingredient. Preferably a daily dose in the range of 1-50 mg/kg is employed. However the daily dose will necessarily be varied depending upon the host treated, the particular route of administration, and the severity of the illness being treated. Accordingly the optimum dosage may be determined by the practitioner who is treating any particular patient.
According to a further aspect of the present invention there is provided a compound of the formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, as defined herein before, for use in a method of treatment of the human or animal body by therapy.
We have found that the compounds defined in the present invention, or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, are effective cell cycle inhibitors (anti-cell proliferation agents), which property is believed to arise from their CDK 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 CDK enzymes, i.e. the compounds may be used to produce a CDK inhibitory effect in a warm-blooded animal in need of such treatment. Thus the compounds of the present invention provide a method for treating the proliferation of malignant cells characterised by inhibition of CDK enzymes, i.e. the compounds may be used to produce an anti-proliferative effect mediated alone or in pail by the inhibition of CDKs. Such a compound of the invention is expected to possess a wide range of anti-cancer properties as CDKs have been implicated in many common human cancers such as leukaemia and breast, lung, colon, rectal, stomach, prostate, bladder, pancreas and ovarian cancer. 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, prostate 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 colon, breast, prostate, lungs and skin. More particularly such compounds of the invention, or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, are expected to inhibit the growth of those primary and recurrent solid tumours which are associated with CDKs, especially those tumours which are significantly dependent on CDKs for their growth and spread, including for example, certain tumours of the colon, breast, prostate, lung, vulva and skin.
It is further expected that a compound of the present invention will possess activity against other cell-proliferation diseases in a wide range of other disease states including leukaemias, fibroproliferative and differentiative disorders, psoriasis, rheumatoid arthritis, Kaposi's sarcoma, haemangioma, acute and chronic nephropathies, atheroma, atherosclerosis, arterial restenosis, autoimmune diseases, acute and chronic inflammation, bone diseases and ocular diseases with retinal vessel proliferation.
Thus according to this aspect of the invention there is provided a compound of the formula (I), or a pharmaceutically acceptable salt or ill vivo hydrolysable ester thereof, as defined herein before, for use as a medicament; and the use of a compound of the formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, as defined herein before, in the manufacture of a medicament for use in the production of a cell cycle inhibitory (anti-cell-proliferation) effect in a warm-blooded animal such as man. Particularly, an inhibitory effect is produced by preventing entry into or progression through the S phase by inhibition of CDK2, CDK4 and/or CDK6, especially CDK2.
According to a further feature of the invention, there is provided a compound of the formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, as defined herein before, in the manufacture of a medicament for use in the treatment of cancers (solid tumours and leukaemias), fibroproliferative and differentiative disorders, psoriasis, rheumatoid arthritis, Kaposi's sarcoma, haemangioma, acute and chronic nephropathies, atheroma, atherosclerosis, arterial restenosis, autoimmune diseases, acute and chronic inflammation, bone diseases and ocular diseases with retinal vessel proliferation, particularly in the treatment of cancers.
According to a further feature of this aspect of the invention there is provided a method for producing a cell cycle inhibitory (anti-cell-proliferation) 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 or in vivo hydrolysable ester thereof, as defined herein before. Particularly, an inhibitory effect is produced by preventing entry into or progression through the S phase by inhibition of CDK2, CDK4 and/or CDK6, especially CDK2.
According to an additional feature of this aspect of the invention there is provided a method of treating cancers (solid tumours and leukaemias), firoproliferative and differentiative disorders, psoriasis, rheumatoid arthritis, Kaposi's sarcoma, haemangioma, acute and chronic nephropathies, atheroma, atherosclerosis, arterial restenosis, autoimmune diseases, acute and chronic inflammation, bone diseases and ocular diseases with retinal vessel proliferation, 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 or in vivo hydrolysable ester thereof, as defined herein before.
Particularly there is provided a method of treating cancer 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 or in vivo hydrolysable ester 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 or in vivo hydrolysable ester thereof, as defined herein before, in association with a pharmaceutically-acceptable diluent or carrier for use in the production of a cell cycle inhibitory (anti-cell-proliferation) 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 or in vivo hydrolysable ester thereof, as defined herein before, in association with a pharmaceutically-acceptable diluent or carrier for use in the treatment of cancers (solid tumours and leukaemias), fibroproliferative and differentiative disorders, psoriasis, rheumatoid arthritis, Kaposi's sarcoma, haemangioma, acute and chronic nephropathies, atheroma, atherosclerosis, arterial restenosis, autoimmune diseases, acute and chronic inflammation, bone diseases and ocular diseases with retinal vessel proliferation, 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 or in vivo hydrolysable ester thereof, as defined herein before, in association with a pharmaceutically-acceptable diluent or carrier for use in the treatment of cancer in a warm-blooded animal such as man.
Preventing cells from entering DNA synthesis by inhibition of essential S-phase initiating activities such as CDK2 initiation may also be useful in protecting normal cells of the body from toxicity of cycle-specific pharmaceutical agents. Inhibition of CDK2 or 4 will prevent progression into the cell cycle in normal cells which could limit the toxicity of cycle-specific pharmaceutical agents which act in S-phase, G2 or mitosis. Such protection may result in the prevention of hair loss normally associated with these agents.
Therefore in a further aspect of the invention there is provided a compound of formula (I) as defined above or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof for use as a cell protective agent.
Therefore in a further aspect of the invention there is provided a compound of formula (I) as defined above or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof for use in preventing hair loss arising from the treatment of malignant conditions with pharmaceutical agents.
Examples of pharmaceutical agents for treating malignant conditions that are known to cause hair loss include alkylating agents such as ifosfamide and cyclophosphamide; antimetabolites such as methotrexate, 5-fluorouracil, gemcitabine and cytarabine; vinca alkaloids and analogues such as vincristine, vinbalstine, vindesine, vinorelbine; taxanes such as paclitaxel and docetaxel; topoisomerase I inhibitors such as irintotecan and topotecan; cytotoxic antibiotics such as doxorubicin, daunorubicin, mitoxantrone, actinomycin-D and mitomycin; and others such as etoposide and tretinoin.
In another aspect of the invention, the compound of formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, may be administered in association with a one or more of the above pharmaceutical agents. In this instance the compound of formula (I) may be administered by systemic or non systemic means. Particularly the compound of formula (I) my may administered by non-systemic means, for example topical administration.
Therefore in an additional feature of the invention, there is provided a method of prevention hair loss during treatment for one or more malignant conditions with pharmaceutical agents, in a warm-blooded animal, such as man, which comprises administering to said animal an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof.
In an additional feature of the invention, there is provided a method of preventing hair loss during treatment for one or more malignant conditions with pharmaceutical agents, in a warm-blooded animal, such as man, which comprises administering to said animal an effective amount of a compound of formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof in simultaneous, sequential or separate administration with an effective amount of said pharmaceutical agent.
According to a further aspect of the invention there is provided a pharmaceutical composition for use in preventing hair loss arising from the treatment of malignant conditions with pharmaceutical agents which comprises a compound of formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, and said pharmaceutical agent, in association with a pharmaceutically acceptable diluent or carrier.
According to a further aspect of the present invention there is provided a kit comprising a compound of formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, and a pharmaceutical agent for treating malignant conditions that is known to cause hair loss.
According to a further aspect of the present invention there is provided a kit comprising:

    a) a compound of formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, in a first unit dosage form;
    b) a pharmaceutical agent for treating malignant conditions that is known to cause hair loss; in a second unit dosage form; and
    c) container means for containing said first and second dosage forms.


According to another feature of the invention there is provided the use of a compound of the formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, in the manufacture of a medicament for the prevention of hair loss during treatment of malignant conditions with pharmaceutical agents.
According to a further aspect of the present invention there is provided a combination treatment for the prevention of hair loss comprising the administration of an effective amount of a compound of the formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, optionally together with a pharmaceutically acceptable diluent or carrier, with the simultaneous, sequential or separate administration of an effective amount of a pharmaceutical agent for treatment of malignant conditions to a warm-blooded animal, such as man.
As stated above the size of the dose required for the therapeutic or prophylactic treatment of a particular cell-proliferation disease will necessarily be varied depending on the host treated, the route of administration and the severity of the illness being treated. A unit dose in the range, for example, 1-100 mg/kg, preferably 1-50 mg/kg is envisaged.
The CDK inhibitory activity defined hereinbefore may be applied as a sole therapy or may involve, in addition to a compound of the invention, one or more other substances and/or treatments. Such conjoint treatment may be achieved by way of the simultaneous, sequential or separate administration of the individual components of the treatment. In the field of medical oncology it is normal practice to use a combination of different forms of treatment to treat each patient with cancer. In medical oncology the other component(s) of such conjoint treatment in addition to the cell cycle inhibitory treatment defined hereinbefore may be: surgery, radiotherapy or chemotherapy. Such chemotherapy may cover three main categories of therapeutic agent:

    (i) other cell cycle inhibitory agents that work by the same or different mechanisms from those defined hereinbefore;
    (ii) cytostatic agents such as antioestrogens (for example tamoxifen, toremifene, raloxifene, droloxifene, iodoxyfene), progestogens (for example megestrol acetate), aromatase inhibitors (for example anastrozole, letrazole, vorazole, exemestane), antiprogestogens, antiandrogens (for example flutamide, nilutamide, bicalutamide, cyproterone acetate), LHRH agonists and antagonists (for example goserelin acetate, luprolide), inhibitors of testosterone 5α-dihydroreductase (for example finasteride), anti-invasion agents (for example metalloproteinase inhibitors like marimastat and inhibitors of urokinase plasminogen activator receptor function) and inhibitors of growth factor function, (such growth factors include for example platelet derived growth factor and hepatocyte growth factor such inhibitors include growth factor antibodies, growth factor receptor antibodies, tyrosine kinase inhibitors and serine/threonine kinase inhibitors); and
    (iii) antiproliferative/antineoplastic drugs and combinations thereof, as used in medical oncology, such as antimetabolites (for example antifolates like methotrexate, fluoropyrimidines like 5-fluorouracil, purine and adenosine analogues, cytosine arabinoside); antitumour antibiotics (for example anthracyclines like doxorubicin daunomycin, epirubicin and idarubicin, mitomyin-C, dactinomycin, mithramycin); platinum derivatives (for example cisplatin, carboplatin); alkylating agents (for example nitrogen mustard, melphalan, chlorambucil, busulphan, cyclophosphamide, ifosfamide, nitrosoureas, thiotepa); antimitotic agents (for example vinca alkaloids like vincristine and taxoids like taxol, taxotere); topoisorerase inhibitors (for example epipodophyllotoxins like etoposide and teniposide, amsacrine, topotecan). According to this aspect of the invention there is provided a pharmaceutical product comprising a compound of the formula (I) as defined hereinbefore and an additional anti-tumour substance as defined hereinbefore for the conjoint treatment of cancer.


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 cell cycle activity 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 illustrated by the following non limiting examples in which, unless stated otherwise:

    (i) temperatures are given in degrees Celsius (° C.); 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; 4.5-30 mmHg) with a bath temperature of up to 60° C.;
    (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 using perdeuterio dimethyl sulphoxide (DMSO-d6) as solvent 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 were run with an electron energy of 70 electron volts in the chemical ionization (CI) mode using a direct exposure probe; where indicated ionization was effected by electron impact (EI), fast atom bombardment (FAB) or electrospray (ESP); values for m/z are given; generally, only ions which indicate the parent mass are reported; and unless otherwise stated, the mass ion quoted is (MH)+;
    (xi) unless stated otherwise compounds containing an asymmetrically substituted carbon and/or sulphur atom have not been resolved;
    (xii) where a synthesis is described as being analogous to that described in a previous example the amounts used are the millimolar ratio equivalents to those used in the previous example;
    (xvi) the following abbreviations have been used:


















THF
tetrahydrofuran;



SM
starting material; and



DCM
dichloromethane.










Example 1
2-{4-[N-(2-Methoxyethy)sulphamoyl]anilino}-4-(2-propyn-1-yloxy)-5-cyanopyrimidine
A solution of 2-chloro-4-(2-propyn-1-yloxy)-5-cyanopyrimidine (Method 17; 219 mg, 1.13 mmol) and 4[N-(2-methoxyethyl)sulphamoyl]aniline (Method 38; 130 mg, 0.56 mmol) in 2-butanol (8 ml) was stirred at 50° C. for 18 hours. Solvent was removed under reduced pressure and the product recrystallized from methanol to give the title compound (145 mg, 67%). NMR: 2.90 (q, 2H), 3.15 (s, 3H), 3.25 (m, 2H), 3.70 (s, 1H), 5.20 (s, 2H), 7.55 (t, 1H), 7.75 (d, 2H), 7.90 (d, 2H), 8.80 (s, 1H); m/z 387.
Examples 2-30
The following compounds were prepared using the procedure of Example 1 using the appropriate pyrimidine and aniline starting materials.















Ex
Compound Name
NMR
m/z
SM
Aniline




















2
2-(4-Ethylsulphonyl-
1.10(t, 3H), 2.10(m, 1H), 2.35(m,
374
Meth
Ref 1



anilino)-4-(2-
1H), 3.20(m, 2H), 3.70-4.00(m,

18



tetrahydrofur-3-yloxy)-5-
4H), 5.70(br s, 1H),



cyanopyrimidine
7.80(d, 2H), 7.95(d, 2H), 7.75(s,




1H)


3
2-{4-[N-(2,2,2-
2.10(m, 1H), 2.30(m, 1H),
443
Meth
Meth



Trifluoroethyl)
3.65(m, 2H), 3.70-4.00(m,

18
44



sulphamoyl]anilino}-4-(2-
4H), 5.70(br s, 1H), 7.80(d,



tetrahydrofur-3-yloxy)-5-
2H), 7.90(d, 2H), 8.45(t, 1H),



cyanopyrimidine
8.75(s, 1H)


4
2-{4-[N-(3-
1.55(m, 2H), 2.10(m, 1H),
433
Meth
Meth



Methoxypropyl)
2.30(m, 1H), 2.75(q, 2H),

18
40



sulphamoyl]anilino}-4-(2-
3.10(s, 3H), 3.25(m, 2H),



tetrahydrofur-3-yloxy)-5-
3.70-3.95(m, 4H), 5.70(br s,



cyanopyrimidine
1H), 7.40(t, 1H), 7.75(d, 2H),




7.90(d, 2H), 8.75(s, 1H)


5
2-{4-[N-(2-Methoxyethyl)
2.10(m, 1H), 2.35(m, 1H),
419
Meth
Meth



sulphamoyl]anilino}-4-(2-
2.90(m, 2H), 3.20(s, 3H), 3.30(t,

18
38



tetrahydrofur-3-yloxy)-5-
2H), 3.75-4.00(m, 4H), 5.70(s,



cyanopyrimidine
1H), 7.55(t, 1H), 7.80(d,




2H), 7.90(d, 2H), 8.75(s, 1H),




10.70(br s, 1H)


6
2-{4-[N-(3-
1.60(m, 2H), 1.65(d, 3H),
415
Meth
Meth



Methoxypropyl)
2.80(m, 2H), 3.20(s, 3H), 3.30(t,

23
40



sulphamoyl]anilino}-4-(3-
2H), 3.70(s, 1H), 5.85(m,



butyn-2-yloxy)-5-
1H), 7.45(t, 1H), 7.80(d, 2H),



cyanopyrimidine
8.00(d, 2H), 8.80(s, 1H),




10.80(br s, 1H)


7
2-{4-(4-Isopropylamino-
1.30(d, 6H), 1.70(br s, 4H),
427
Meth
Meth



butylsulphonyl)anilino}-
2.90(br s, 2H), 3.25(m, 1H),

17
49



4-(2-propyn-1-yloxy)-5-
3.40(m, 2H), 3.80(s, 1H), 5.20(s,



cyanopyrimidine
1H), 7.90(d, 2H), 8.10(d,




2H), 8.50(brs, 1H), 8.85(s,




1H), 10.90(s, 1H)


8
2-{4-[N-(3-
1.60(m, 2H), 2.75(m, 2H),
401
Meth
Meth



Methoxypropyl)
3.10(s, 3H), 3.25(t, 2H), 3.70(s,

17
40



sulphamoyl]anilino}-4-(2-
1H), 5.15(s, 2H), 7.40(t,



propyn-1-yloxy)-5-
1H), 7.70(d, 2H), 7.90(d, 2H),



cyanopyrimidine
8.80(s, 1H)


9
2-(4-Ethylsulphonyl-
1.10(t, 3H), 3.25(q, 2H), 3.70(s,
342
Meth
Ref 1



anilino)-4-(2-propyn-1-
1H), 5.20(s, 2H), 7.80(d,

17



yloxy)-5-cyanopyrimidine
2H), 8.00(d, 2H), 8.80(s, 1H)


10
2-{4-[N-(2,2,2-
3.65(m, 2H), 3.70(s, 1H), 5.20(s,
411
Meth
Meth



Trifluoroethyl)
2H), 7.80(d, 2H), 7.95(d,

17
44



sulphamoyl]anilino}-4-(2-
2H), 8.45(t, 1H), 8.80(s, 1H)



propyn-1-yloxy)-5-



cyanopyrimidine


11
2-[4-(N-Allyl
3.40(m, 2H), 3.70(s, 1H), 5.00(d,
369
Meth
Meth



sulphamoyl)anilino]-4-(2-
1H), 5.10(d, 1H), 5.20(s,

17
42



propyn-1-yloxy)-5-
2H), 5.65(m, 1H), 7.65(t, 1H),



cyanopyrimidine
7.75(d, 2H), 7.90(d, 2H), 8.80(s, 1H)


12
2-{4-[N-(3-
1.55(m, 2H), 1.85(s, 3H), 2.75(q,
415
Meth
Meth



Methoxypropyl)
2H), 3.10(s, 3H), 3.25(m,

20
40



sulphamoyl]anilino}-4-(2-
2H), 5.15(m, 2H), 7.40(t, 1H),



butyn-1-yloxy)-5-
7.70(d, 2H), 7.90(d, 2H), 8.80(s,



cyanopyrimidine
1H)


13
2-{4-[N-(3-
1.55(m, 2H), 1.70(m, 1H),
447
Meth
Meth



Methoxypropyl)
2.00(m, 1H), 2.75(m, 2H),

19
40



sulphamoyl]anilino}-4-
3.10(s, 3H), 3.25(m, 2H), 3.50(m,



(tetrahydrofur-3-
1H), 3.65(m, 1H), 3.75(m,



ylmethoxy)-5-
3H), 4.40(m, 2H), 7.40(t, 1H),



cyanopyrimidine
7.70(d, 2H), 7.90(d, 2H), 8.70(s, 1H)


14
2-{4-[N-(3-
1.55(m, 2H), 2.70(m, 4H),
415
Meth
Meth



Methoxypropyl)
2.90(s, 1H), 3.10(s, 3H), 3.25(m,

21
40



sulphamoyl]anilino}-4-(3-
2H), 4.55(t, 2H), 7.40(t,



butyn-1-yloxy)-5-
1H), 7.70(d, 2H), 7.90(d, 2H),



cyanopyrimidine
8.75(s, 1H)


15
2-{4-[N-(3-
1.60(m, 2H), 2.45(m, 4H),
476
Meth
Meth



Methoxypropyl)
2.70(m, 4H), 3.15(s, 3H), 3.25(m,

24
40



sulphamoyl]anilino}-4-(2-
2H), 3.50(m, 4H), 4.60(m,



morpholinoethoxy)-5-
2H), 7.45(t, 1H), 7.75(d, 2H),



cyanopyrimidine
7.90(d, 2H), 8.75(s, 1H),




10.75(br s, 1H)


16
2-{4-[N-(3-
1.60(m, 2H), 2.75(q, 2H),
465
Meth
Meth



Methoxypropyl)
3.10(s, 3H), 3.20(m, 5H), 3.45(m,

30
40



sulphamoyl]anilino}-4-[2-
2H), 3.60(m, 2H), 3.80(m,



(2-methoxyethoxy)
2H), 4.60(m, 2H), 7.40(t, 1H),



ethoxy]-5-
7.70(d, 2H), 7.90(d, 2H), 8.75(s,



cyanopyrimidine
1H)


17
2-{4-[N-(3-
1.60(m, 2H), 2.75(q, 2H),
421
Meth
Meth



Methoxypropyl)
3.10(s, 3H), 3.25(t, 2H), 3.30(s,

22
40



sulphamoyl]anilino}-4-(2-
3H), 3.70(m, 2H), 4.60(m,



methoxyethoxy)-5-
2H), 7.40(t, 1H), 7.70(d, 2H),



cyanopyrimidine
7.90(d, 2H), 8.75(s, 1H)


18
2-{4-[N-(3-
0.40(m, 2H), 0.60(m, 2H),
417
Meth
Meth



Methoxypropyl)
1.30(m, 1H), 1.60(m, 2H),

25
40



sulphamoyl]anilino}-4-
2.75(m, 2H), 3.10(s, 3H), 3.25(t,



(cyclopropylmethoxy)-5-
2H), 4.30(d, 2H), 7.40(t,



cyanopyrimidine
1H), 7.75(d, 2H), 7.90(d, 2H),




8.75(s, 1H), 10.65(br s, 1H)


19
2-{4-[N-(3-
1.35(t, 3H), 1.55(m, 2H), 2.75(m,
391
Meth
Meth



Methoxypropyl)
2H), 3.10(s, 3H), 3.25(m,

31
40



sulphamoyl]anilino}-4-
2H), 4.50(m, 2H), 7.40(t, 1H),



ethoxy-5-cyanopyrimidine
7.70(d, 2H), 7.90(d, 2H), 8.70(s, 1H)


20
2-{4-[N-(2,2,2-
1.40(d, 6H), 3.65(m, 2H),
415
Meth
Meth



Trifluoroethyl)
5.40(m, 1H), 7.80(d, 2H),

26
44



sulphamoyl]anilino}-4-
7.90(d, 2H), 8.45(t, 1H), 8.70(s,



isopropoxy-5-
1H)



cyanopyrimidine


21
2-{4-[N-(3-
1.40(d, 6H), 1.55(m, 2H),
405
Meth
Meth



Methoxypropyl)
2.75(q, 2H), 3.10(s, 3H), 3.25(m,

26
40



sulphamoyl]anilino}-4-
2H), 5.25(m, 1H), 7.40(t,



isopropoxy-5-
1H), 7.70(d, 2H), 7.90(d, 2H),



cyanopyrimidine
8.70(s, 1H)


22
2-{4-[N-(2-Methoxyethyl)
1.40(d, 6H), 2.85(q, 2H), 3.15(s,
391
Meth
Meth



sulphamoyl]anilino}-4-
3H), 3.25(t, 2H), 5.45(m,

26
38



isopropoxy-5-
1H), 7.55(t, 1H), 7.70(d, 2H),



cyanopyrimidine
7.90(d, 2H), 8.70(s, 1H)


23
2-{4-[N-(3-
1.60(m, 2H), 2.80(q, 2H),
377
Meth
Meth



Methoxypropyl)
3.20(s, 3H), 3.30(m, 2H), 4.10(s,

27
40



sulphamoyl]anilino}-4-
3H), 7.45(t, 1H), 7.80(d,



methoxy-5-
2H), 8.00(d, 2H), 8.75(s, 1H),



cyanopyrimidine
10.70(s, 1H)


24
2-{4-[N-(2-Methoxyethyl)
0.90(t, 3H), 1.40(m, 2H), 1.75(m,
405
Meth
Meth



sulphamoyl]anilino}-4-
2H), 2.85(q, 2H), 3.15(s,

28
28



butoxy-5-
3H), 3.30(t, 2H), 4.45(t, 2H),



cyanopyrimidine
7.50(t, 1H), 7.70(d, 2H), 7.90(d,




2H), 8.70(s, 1H)


25
2-{4-[N-(3-
0.85(t, 3H), 1.40(m, 2H), 1.55(m,
435
Meth
Meth



Methoxypropyl)
2H), 1.65(m, 2H), 2.75(q,

32
40



sulphamoyl]anilino}-4-
2H), 3.10(s, 3H), 3.30(m,



butylthio-5-
4H), 7.40(t, 1H), 7.70(d, 1H),



cyanopyrimidine
7.90(d, 1H), 8.65(s, 1H)


26
2-{4-[N-(2-Methoxyethyl)
0.90(t, 3H), 1.40(m, 2H), 1.65(m,
421
Meth
Meth



sulphamoyl]anilino}-4-
2H), 2.90(q, 2H), 3.10(s,

32
38



butylthio-5-
3H), 3.30(m, 4H), 7.55(t, 1H),



cyanopyrimidine
7.70(d, 2H), 7.90(d, 2H), 8.60(s, 1H)


27
2-{4-[N-(Tetrahydrofur-2-
1.50(m, 1H), 1.70-1.90(br m,
413
Meth
Meth



ylmethyl)sulphamoyl]
3H), 2.75(t, 2H), 3.55(m, 1H),

17
39



anilino}-4-(2-propyn-1-
3.65(m, 1H), 3.70(m, 1H),



yloxy)-5-cyanopyrimidine
3.80(m, 1H), 5.20(d, 2H),




7.55(t, 1H), 7.55(d, 2H), 7.95(d, 2H),




8.80(s, 1H)


28
2-{4-[N-(Cyclopropyl)
0.40(m, 2H), 0.50(m, 2H),
369
Meth
Meth



sulphamoyl]anilino}-4-(2-
2.10(m, 1H), 3.70(s, 1H), 5.10(d,

17
41



propyn-1-yloxy)-5-
2H), 7.60(m, 3H), 7.95(d,



cyanopyrimidine
2H), 8.45(s, 1H)


29
2-[4-(N,N-Dimethyl
2.60(s, 6H), 3.70(s, 1H), 5.20(d,
357
Meth
Com av



sulphamoyl)anilino]-4-(2-
2H), 7.70(d, 2H), 8.00(d,

17



propyn-1-yloxy)-5-
2H), 8.80(s, 1H)



cyanopyrimidine


30
2-(4-Sulphamoyl anilino)-
3.70(s, 1H), 5.15(s, 2H), 7.20(s,
329
Meth
Com av



4-(2-propyn-1-yloxy)-5-
2H), 7.75(d, 2H), 7.90(d,

17



cyano pyrimidine
2H), 8.80(s, 1H)


31
2-(4-Ethylsulphonyl
1.10(t, 3H), 1.35(d, 6H), 3.23(q,
346
Meth
Ref 1



anilino)-4-isopropoxy-5-
2H), 5.45(m, 1H), 7.83(d,

26



cyanopyrimidine
2H), 7.95(d, 2H), 8.70(s, 1H)


32
2-[4-(N-Methyl
2.40(d, 3H), 3.70(m, 1H),
343
Meth
Ref 2



sulphamoyl)anilino]-4-(2-
5.20(m, 2H), 7.28(q, 1H),

17



propyn-1-yloxy)-5-
7.72(d, 2H), 7.95(d, 2H), 8.80(s,



cyanopyrimidine
1H)


33
2-{4-[N-Methyl-N-(2-
2.70(s, 3H), 3.10(t, 2H), 3.20(s,
401
Meth
Ref 3



methoxyethyl)
3H), 3.43(t, 2H), 3.73(m,

17



sulphamoyl]anilino}-4-(2-
1H), 5.18(s, 2H), 7.74(d, 2H),



propyn-1-yloxy)-5-cyano
8.00(d, 2H), 8.80(s, 1H)



pyrimidine


34
2-[4-(N-Cyclobutyl
1.45(m, 2H), 1.70(m, 2H),
483
Meth
Meth



sulphamoyl)anilino]-4-(2-
1.90(m, 2H), 3.60(m, 1H),

17
54



propyn-1-yloxy)-5-
3.70(m, 1H), 5.17(m, 2H),



cyanopyrimidine
7.70(d, 2H), 7.80(d, 1H), 7.90(d,




2H), 8.80(s, 1H)


35
2-{4-[N-(Cyclopropyl
0.10(m, 2H), 0.40(m, 2H),
383
Meth
Meth



methyl)sulphamoyl]
0.83(m, 1H), 2.68(t, 2H), 3.75(t,

17
55



anilino}-4-(2-propyn-1-
1H), 5.20(d, 2H), 7.60(t,



yloxy)-5-cyanopyrimidine
1H), 7.80(d, 2H), 7.95(d, 2H),




8.83(s, 1H)


36
2-{4-[N-(3-Methoxy
0.95(t, 3H), 1.35(d, 3H), 1.58(m,
419
Meth
Ref 4



propyl)sulphamoyl]
2H), 1.72(m, 2H), 2.75(q,

16



anilino}-4-(but-2-yloxy)-
2H), 3.15(s, 3H), 3.25(t, 2H),



5-cyanopyrimidine
5.25(m, 1H), 7.40(t, 1H), 7.72(d,




2H), 7.90(d, 2H), 8.70(s,




1H)





Ref 1: Helv. Chim. Acta, 66(4), 1046-52, (1983) 


Ref 2: J. Med. Chem, 40(11), 1619-1633, (1997) 


Ref 3 Method 16 WO 02/04429 


Ref 4 Method 4 WO 02/04429 






Example 37
2-{4-[N-(2-Piperidinoethyl)sulphamoyl]anilino}-4-isopropoxy-5-cyanopyrimidine
To a stirred suspension of 2-chloro-4-isopropoxy-5-cyanopyrimidine (Method 26; 505 mg, 2.56 mmol) and 4-[N-(2-piperidinoethyl)sulphamoyl]aniline (Method 43; 362 mg, 1.28 mmol) in 2-butanol (10 ml) was added 1M HCl in ether (3.6 ml, 3.6 mmol). The reaction was warmed to 70° C. and stirred for 18 hours. The reaction was filtered and the solvent removed under reduced pressure to yield a yellow solid. This solid was partitioned between DCM (40 ml) and 2M sodium hydroxide (40 ml). The phases were separated and the aqueous layer was extracted with DCM (30 ml). The organic phases were combined and washed with water (30 ml) and brine (30 ml). Solvent was removed under reduced pressure and the resulting solid was recrystallized from methanol to give the title compound (15 mg). NMR: 1.35 (m, 2H), 1.45 (m, 10H), 2.25 (m, 6H), 2.85 (m, 2H), 5.45 (m, 1H), 7.30 (br s, 1H), 7.80 (d, 2H), 7.90 (d, 2H),8.75 (s, 1H), 10.60 (br s, 1H); m/z 444.
Example 38
2-{4-[N-(2,2,2-Trifluortoethyl)sulplamoyl]anilino}-4-(2-methoxyethoxy)-5-cyanopyrmidine
To a solution of 2-{4-[N-(2,2,2-trifluoroethyl)sulphamoyl]anilino}-4-(4-chlorophenoxy)-5-cyanopyrimidine (Method 35; 386 mg, 0.8mmol) in 2-methoxyethanol (6 ml) was added 1M HCl in ether (1.6 ml, 1.6 mmol). The reaction was the stirred at 115° C. for 48 hours Solvent was removed under reduced pressure and the product recrystallised from methanol to give the title compound (152 mg). NMR: 3.30 (s, 3H), 3.60-3.80 (m, 4H), 4.60 (t, 2H), 7.80 (d, 2H), 7.90 (d, 2H), 8.45 (t, 1H), 8.75 (s, 1H); m/z 431.
Examples 39-40
The following compounds were prepared using the procedure of Example 38 and the appropriate starting material.














Ex
Compound Name
NMR
m/z
SM







39
2-{4-[N-(2-Methoxyethyl)
2.85(q, 2H), 3.15(s, 3H), 3.25(m, 5H),
407
Meth



sulphamoyl]anilino}-4-(2-
3.70(m, 2H), 4.60(m, 2H), 7.50(t, 1H),

37



methoxyethoxy)-5-
7.75(d, 2H), 7.90(d, 2H), 8.75(s, 1H)



cyanopyrimidine


40
2-(4-ethylsulphonyl anilino)-
1.10(t, 3H), 3.20-3.40(m, 5H), 3.75(d,
362
Meth



4-(2-methoxy ethoxy)-5-
2H), 4.65(d, 2H), 7.85(d, 2H), 8.00(d,

36



cyanopyrimidine
2H), 8.80(s, 1H), 10.80(br s, 1H)









Preparation of Starting Materials

The starting materials for the above Examples are either commercially available or are readily prepared by standard methods from known materials. For example the following reactions are illustrations but not limitations of the preparation of some of the starting materials used in the above reactions.
Method 1
2-Chloro-4-(2-propyn-1-yloxy)-5-N-t-butylcarbamoylpyrimidine
Sodium hydride (60% suspension in mineral oil, 720 mg, 0.018 mol) was added to a solution of propargyl alcohol (0.70 ml, 0.012 mol) in dry THF (70 ml). When effervescence had stopped the resulting suspension was slowly added to a solution of 5-N-t-butylcarbamoyl-2,4-dichloropyrimidine (Method 51; 3.00 g, 0.012 mol) in dry THF (130 ml). The reaction was stirred at room temperature for 18 hours. Solvent was removed under reduced pressure and the resulting solid was triturated with ether (50 ml). The suspension was filtered and the ether filtrate was washed with citric acid (25 ml), water (25 ml) and brine (25 ml) then dried. Removal of solvent gave the title compound (3.09 g, 96%). NMR: 1.30 (s, 10H), 3.70 (m, 1H), 5.10 (d, 2H), 7.80 (br s, 1H), 8.65 (s, 1H); m/z 268
Methods 2-16
Using the procedure of method 1 and using the appropriate alcohols or thiols the following compounds were prepared. (N.B. Method 5 was used without purification).














Meth
Compound Name
m/z





2
2-Chloro-4-(tetrahydrofur-3-yloxy)-5-N-t-butylcarbamoylpyrimidine
299


3
2-Chloro-4-(tetrahydrofur-3-ylmethoxy)-5-N-t-butylcarbamoylpyrimidine
313


4
2-Chloro-4-(2-butyn-1-yloxy)-5-N-t-butylcarbamoylpyrimidine
281


5
2-Chloro-4-(3-butyn-1-yloxy)-5-N-t-butylcarbamoylpyrimidine


6
2-Chloro-4-(2-methoxyethoxy)-5-N-t-butylcarbamoylpyrimidine
287













Meth
Compound Name
NMR
m/z





7
2-Chloro-4-(3-butyn-2-yloxy)-5-
1.35(s, 9H), 1.60(m, 3H), 3.65(s, 1H),
281



N-t-butylcarbamoylpyrimidine
5.70(m, 1H), 7.70(m, 1H), 8.65(s, 1H)


8
2-Chloro-4-(2-
1.35(s, 9H), 2.45(m, 4H), 2.70(t, 2H),
342



morpholinoethoxy)-5-N-t-
3.55(m, 4H), 4.50(t, 2H), 7.75(br s, 1H),



butylcarbamoylpyrimidine
8.65(s, 1H)


9
2-Chloro-4-[2-(2-
1.35(s, 9H), 3.20(s, 3H), 3.45(m, 2H),
331



methoxyethoxy)ethoxy]-5-N-t-
3.55(m, 2H), 3.80(m, 2H), 4.50(m, 2H),



butylcarbamoylpyrimidine
7.70(br s, 1H), 8.70(s, 1H)


10 
2-Chloro-4-
0.45(m, 2H), 0.65(m, 2H), 1.30(m, 1H),
283



(cyclopropylmethoxy)-5-N-t-
1.40(s, 9H), 4.30(d, 2H), 7.80(br s, 1H),



butylcarbamoylpyrimidine
8.40(s, 1H)


11 
2-Chloro-4-ethoxy-5-N-t-
1.35(m, 12H), 4.45(q, 2H), 7.75(br s,
257



butylcarbamoylpyrimidine
1H), 8.60(s, 1H)


12 
2-Chloro-4-isopropoxy-5-N-t-
1.35(m, 15H), 5.30(m, 1H), 7.65(br s,
271



butylcarbamoylpyrimidine
1H), 8.60(s, 1H)


13 
2-Chloro-4-methoxy-5-N-t-
1.35(s, 9H), 4.00(s, 3H), 7.75(br s, 1H),



butylcarbamoylpyrimidine
8.60(s, 1H)


14 
2-Chloro-4-butoxy-5-N-t-
0.90(t, 3H), 1.35(s, 9H), 1.45(m, 2H),
285



butylcarbamoylpyrimidine
1.70(m, 2H), 4.40(t, 2H), 7.75(br s, 1H),




8.60(s, 1H)


15 
2-Chloro-4-butylthio-5-N-t-
0.90(t, 3H), 1.30-1.45(br m, 11H), 1.60(m,
301



butylcarbamoylpyrimidine
2H), 3.10(t, 2H), 8.15(br s, 1H), 8.40(s, 1H)


16 
2-Chloro-4-but-2-yloxy-5-N-t-
0.83(t, 3H), 1.32(m, 12H), 1.70(m, 2H),
285



butylcarbamoylpyrimidine
5.20(m, 1H), 7.70(br s, 1H), 8.60(s, 1H)









Method 17

2-Chloro-4-(2-propyn-1-yloxy)-5-cyanopyrimidine

A solution of 2-chloro-4-(2-propyn-1-yloxy)-5-N-t-butylcarbamoylpyrimdine (Method 1; 3.49 g, 0.013 mol) in thionyl chloide (135 ml) was heated at 95° C. for 18 hours. Solvent was removed under reduced pressure and the resulting brown oil dissolved in ether. Ether was removed under reduced pressure to give the title compound as an oil (3 g) which was used crude in subsequent reactions. NMR: 3.75 (t, 1H), 5.15 (d, 2H), 5.45 (br s, 2H), 9.05 (s, 1H); m/z 193.
Methods 18-32
Using the procedure of Method 17 the following compounds were prepared. In most cases purity was checked by HPLC and the product was used crude in the next reaction.














Meth
Compound Name
SM





18
2-Chloro-4-(tetrahydrofur-3-yloxy)-5-cyanopyrimidine
Meth 2


19
2-Chloro-4-(tetrahydrofur-3-ylmethoxy)-5-
Meth 3



cyanopyrimidine


20
2-Chloro-4-(2-butyn-1-yloxy)-5-cyanopyrimidine
Meth 4


21
2-Chloro-4-(3-butyn-1-yloxy)-5-cyanopyrimidine
Meth 5


22
2-Chloro-4-(2-methoxyethoxy)-5-cyanopyrimidine
Meth 6


23
2-Chloro-4-(3-butyn-2-yloxy)-5-cyanopyrimidine
Meth 7


24
2-Chloro-4-(2-morpholinoethoxy)-5-cyanopyrimidine
Meth 8


25
2-Chloro-4-(cyclopropylmethoxy)-5-cyanopyrimidine
Meth 10


26
2-Chloro-4-isopropoxy-5-cyanopyrimidine
Meth 12


27
2-Chloro-4-methoxy-5-cyanopyrimidine
Meth 13


28
2-Chloro-4-butoxy-5-cyanopyrimidine
Meth 14


29
2-Chloro-4-but-2-yloxy-5-cyanopyrimidine
Meth 16













Meth
Compound Name
m/z
SM





30
2-Chloro-4-[2-(2-methoxyethoxy)ethoxy]-
257
Meth 9



5-cyanopyrimidine













Meth
Compound Name
NMR
SM





31
2-Chloro-4-ethoxy-5-
1.35(t, 3H), 4.50(q,
Method



cyanopyrimidine
2H), 9.00(s, 1H)
11


32
2-Chloro-4-butylthio-5-
0.90(t, 3H), 1.40(m, 3H),
Method



cyanopyrimidine
1.65(m, 3H), 3.25(t,
15




2H), 8.90(s, 1H)









Method 33

2-Chloro-4-(4-chlorophenoxy)-5-N-t-butylcarbamoylpyrimidine

To a solution of 4-chlorophenol (338 mg, 2.63 mmol) in dry THF (10 ml) was added sodium hydride, (105 mg, 2.63 mmol). When effervescence had stopped this solution was slowly added to a solution of 2,4-dichloro-5-N-t-butylcarbamoylpyrimidine (Method 51; 680 mg, 2.74 mmol) in dry THF (15 ml). The reaction was stirred at room temperature for 2 hours. Solvent was removed under reduced pressure and the resulting solid was suspended in diethyl ether (50 ml). The solid was filtered and the filtrate was washed with 2M sodium hydroxide (30 ml), citric acid (30 ml), water (30 ml) and brine (30 ml) before drying. Removal of the ether gave the title compound (880 mg, 99%). M/z 339.
Method 34
2-Chloro-4-(4-chlorophenoxy)-5-cyanopyrimidine
A solution of 2-chloro-4-(4-chlorophenoxy)-5-N-t-butylcarbamoylpyrimidine (Method 33; 200 mg, 0.59 mmol) in thionyl chloride (5 ml, 69 mmol)-was heated at 95° C. for 18 hours. The thionyl chloride was removed under reduced pressure and the resulting brown oil was dissolved in diethyl ether (25 ml). The ether was removed under reduced pressure to give the title compound (210 mg). NMR: 7.35 (d, 2H), 7.55 (d, 2H), 9.20 (s, 1H).
Method 35
2-{4-[N-(2,2,2-Trifluorosethyl)sulphamoyl]anilino}-4-(4-chlorohenoxy)-5-cyanopyrimidine
A suspension of 2-chloro-4-(4-chlorophenoxy)-5-cyanopyrimidine (Method 34; 500 mg, 1.88 mmol)and 4-[N-(2,2,2-trifluoroethyl)sulphamoyl]aniline (Method 44; 432 mg, 1.70 mmol) in 2-butanol (10 ml) was stirred at 45° C. for 18 hours. Solvent was removed under reduced pressure and the product triturated with ether. The resulting solid was collected by filtration and washed with ether to give the title compound (480 mg) which was used crude in subsequent reactions. M/z 483.
Methods 36-37
Following the procedure of method 35 and using the appropriate aniline starting material the following compounds were prepared.












Meth
Compound Name
m/z







36
2-(4-Ethylsulphonylanilino)-4-(4-
414



chlorophenoxy)-5-cyanopyrimidine


 371
2-{4-[N-(2-Methoxyethyl)sulphamoyl]anilino}-
459



4-(4-chlorophenoxy)-5-



cyanopyrimidine





1Aniline: Method 38. 






Method 38

4-[N-(2-Methoxyethyl)sulphamoyl]aniline

A mixture of 2-methoxyethylamine (859 mg, 11.4 mmol), sulphanilyl fluoride (1.0 g, 5.71 mmol) and triethylamine (1.72 g, 22.9 mmol) in 1-butanol (15 ml) was heated at reflux for 18 hours. The mixture was allowed to cool and the volatiles were removed by evaporation. The residue was purified by chromatography eluting with ethyl acetate/hexane (50:50) increasing in polarity to (70:30) to give the title compound (860 mg, 65%). NMR: 2.78 (q, 2H), 3.15 (s, 3H), 3.25 (t, 2H), 5.87 (s, 2H), 6.58 (d, 2H), 7.10 (t, 1H), 7.40 (d, 2H); m/z 231.
Method 39-44
The following compounds were prepared using the procedure of Method 38.














Meth
Compound Name
m/z





39
4-[N-(Tetrahydrofur-2-ylmethyl)sulphamoyl]aniline
257













Meth
Compound Name
NMR
m/z





40
4-[N-(3-Methoxypropyl)
1.60(m, 2H), 2.95(q, 2H), 3.20(s, 3H), 3.30(t,
244



sulphamoyl]aniline
2H), 4.10(br s, 2H), 4.90(br t, 1H), 6.60(d, 2H),




7.55(d, 2H)


41
4-(N-Cyclopropyl-
0.01-0.15(m, 4H), 1.70-1.75(m, 1H), 5.60(s,
230



sulphamoyl)aniline
2H), 6.30(d, 2H), 7.05(s, 1H), 7.10(d, 2H)


42
4-(N-Allylsulphamoyl)
3.30(t, 2H), 5.00(d, 1H), 5.10(d, 1H), 5.65(m,
212



aniline
1H), 5.85(s, 2H), 6.40(d, 2H), 7.20(t, 1H), 7.40(d, 2H)


43
4-[N-(2-Piperidinoethyl)
1.30(br m, 2H), 1.40(br m, 4H), 2.25(br m, 6H),
283



sulphamoyl]aniline
2.70(br t, 2H), 5.85(s, 2H), 6.60(d, 2H), 6.80(br s, 1H),




7.40(d, 2H)


44
4-[N-(2,2,2-
3.50(m, 2H), 5.90(br s, 2H), 6.60(d, 2H), 7.40(d,
254



Trifluoroethyl)
2H), 8.00(br t, 1H)



sulphamoyl]aniline









Method 45

1-Nitro-4-[4-(phthalimido)butylsulphide]phenyl

4-Nitrothiophenol (1.9 g, 12.26 mmol) was stirred in DMF under nitrogen and cooled (ice/water bath). Sodium hydride (60% dispersion in oil: 400 mg, 10 mmol) was added in portions and the mixture stirred for 10 minutes. N-(4-bromobulty)phthalimide (2.8 g, 9.93 mmol) in DMF (10 ml) was added and the reaction was heated at 90° C. for 1.5 hours, cooled to room temperature and stood overnight at ambient temperature. Volatiles were removed by evaporation, water (20 ml) was added and the solution was extracted into ethyl acetate (50+25 ml). The organic layers were combined, washed with water (20 ml) and brine (20 ml), dried and the volatiles evaporated. The resulting gum was triturated twice with isohexane. The solvent was decanted to give the title compound as a solid (3.8 g). NMR: 1.64 (m, 2H), 1.74 (m, 2H), 3.12 (t 2H), 3.60 (t, 2H), 7.45 (d, 2H), 7.81 (m, 41H), 8.06 (d, 2H)
Method 46
1-Nitro-4-[4-(phthalimido)butylsulohonyl]phenyl
A solution of chromium(VI)oxide (3.5 g 35.0 mmol) in water (3 ml) and glacial acetic acid (12.5 ml) was added dropwise over 15 to 20 minutes to a solution of 1-nitro-4-[4-phthalimido)butylsulphide]phenyl (Method 45; 3.5 g, 9.83 mmol) in glacial acetic acid (17.5 ml) heated at 90-100° C. The mixture was-then heated at 100° C. for 3.5 hours. The reaction was cooled, poured onto crushed ice (250 g), and the solid was collected by filtration and washed with water. The solid was dried by azeotroping with methanol 3 times to give the title compound (3.4 g). m/z 389.
Method 47
1-Nitro-4-(4-aminobutylsulphonyl)phenyl
1-Nitro-4-[4-(phthalimido)butylsulphonyl]phenyl (Method 46; 3 g 7.73 mmol) was heated at 90° C. in acetonitrile (30 ml) and methanol (10 ml). Hydrazine hydrate(0.76 ml, 15.7 mmol) was added and the reaction was heated for 1.5 hours, then cooled, and stood overnight at ambient temperature. The resulting solid was removed by filtration and washed with methanol. The combined filtrates were evaporated to give the title compound (2.3 g). m/z 259.
Method 48
1-Nitro-4-[4-(isopropylamino)butylsulphonyl]phenyl
1-Nitro-4-(4-aminobutylsulphonyl)phenyl (Method 47; 2 g, 7.75 mmol) was stirred in methanol (20 ml) and acetone (2.3 ml) was added. Sodium cyanoborohydride (730 mg, 11.62 mmol) was added in portions over 5 minutes and the reaction was stirred for 2.5 hours. Water (15 ml) was added, and the organic solvents were removed by evaporation. Water (20 ml) was added and the solution was extracted with ethyl acetate (2×60 ml). The combined organic layers were washed with water (25 ml) and brine (25 ml). The volatiles were evaporated and the residue was purified by chromatography on neutral alumina (activity II), eluting with DCM increasing polarity to methanol: DCM (3:97) to give the title compound (1.26 g). m/z 301
Method 49
4-[4-(Isopropylamino)butylsulphonyl]aniline
1-Nitro-4-[4-(isopropylamino)butylsulphonyl]phenyl Method 48; 1.2 g 4 mmol) was dissolved in ethanol (20 ml) and 1M hydrochloric acid and hydrogenated at 1 atmosphere over 10% palladium on carbon for 4 hours. The catalyst was removed by filtration and the volatiles evaporated to give the title compound as a foam m/z 271.
Method 50
2,4-Dichloro-5-chloroformylpyrimidine
5-Carboxy-2,4-dihydroxypyrimidine (19.0 g, 0.12 mol), phosphorous pentachloride (83.5 g, 0.40 mol); and phosphoryl chloride (28.3 ml, 0.30 mol) were heated at 114° C. for 5 hours. The resulting solution was cooled overnight and the volatiles removed by evaporation. The residue was purified by vacuum distillation to yield the title compound as a clear oil (17.85 g, 70%). M/z 211.
Method 51
2,4-Dichloro-5-N-t-butylcarbamoylprimidine
A solution of 2,4-dichloro-5-chloroformylpyrimidine (Method 50; 9.8 g, 46.3 mmol) in dry THF (50 ml) was cooled to −15° C. t-Butylamine (5.2 ml, 49.3 mmol) and triethylamine (6.9 ml, 49.5 mmol) in dry THF (20 ml) were added slowly so as to maintain the temperature below −10° C. The resulting mixture was stirred at −10° C. for 2 hours, allowed to warm to ambient temperature and stirred for a further 30 minutes. The precipitate was removed by filtration and the filtrate evaporated to give a viscous oil. High vacuum evaporation gave a solid (10.48 g, 90%). NMR: 1.49 (s, 9H), 6.19 (brs, 1H), 8.86 (s, 1H); m/z 248.
Method 52
4-(N-Cyclobutylsulphamoyl)nitrobenzene
A stirred solution of triethylamine (7.8 ml, 0.056 mol) and cyclobutylamine (4.8 ml, 0.056 mol) in DCM (100 ml) was cooled in an ice/water bath. A solution of 4-nitrobenzene sulphonyl chloride (11.3 g, 0.05 mol) in DCM (100 ml) was added dropwise. The reaction was stirred for one hour the cooling bath was removed and the mixture stirred for further 3 hours. The reaction mixture was washed with 1M hydrochloric acid (80 ml), water (50 ml) and then brine. The volatiles were evaporated to give the title compound (11.79 g). NMR: 1.50 (m, 2H), 1.75 (m, 2H), 1.9 (m, 2H), 3.70 (m, 1H), 8.00 (1, 2H), 8.30 (d, 1H), 8.40 (d, 2H); m/z: 256.
Method 53
Using the procedure of Method 52 starting from 4-nitrobenzenesulphonyl chloride and the appropriate commercially available amine the following compounds were prepared.













Meth
Compound Name
NMR
m/z







53
4-[N-(Cyclopropylmethyl)
0.10(m, 2H), 0.35(m, 
256



sulphamoyl]nitrobenzene
2H), 0.83(m, 1H), 2.75(t, 




2H), 8.05(d, 2H), 8.13(m,




1H), 8.45(d, 2H)









Method 54

4-(N-Cyclobutylsulphamoyl)aniline

4-(N-Cyclobutylsulphamoyl)nitrobenzene (Method 52; 11.79 g) was reduced by hydrogenation in ethanol (300 ml) over 10% palladium on carbon (1 g) at 3 bar pressure. The catalyst was removed by filtration and the volatiles evaporated to give the title compound (7.84 g). NMR: 1.50 (m, 2H), 1.75 (m, 2H), 1.93 (m, 2H), 3.60 (m, 1H), 5.90 (br s, 2H), 6.65 (d, 2H), 7.45 (d, 2H); m/z: 226.
Method 55
4-[N-(Cycloproplymethyl)sulphamoyl]aniline
4-[N-(Cyclopropylmethyl)sulphamoyl]nitrobenzene (Method 53; 12.58 g) was reduced by hydrogenation in ethanol (300 ml) over 10% palladium on carbon (1 g) at 3 bar pressure. The catalyst was removed by filtration and the volatiles evaporated. 2 g of this crude product was purified by chromatography eluting with DCM increasing in polarity to 10% MeOH/DCM to give the title compound (850 mg). NMR: 0.08 (m, 2H), 0.25 (m, 2H), 0.83 (m, 1H, 2.80 (t, 2H), 4.15 (br s, 2H), 4.50 (br t, 1H), 6.65 (d, 2H), 7.62 (d, 2H); m/z: 226.

## Claims

1. A compound of formula (I): 




wherein:
X is —O— or —S—; 
R1 is selected from halo, nitro, cyano, hydroxy, amino, carboxy, carbamoyl, mercapto, C1-6alkyl, C2-6alkenyl and C2-6alkynyl; 
p is 0-4; wherein the values of R1 may be the same or different; 
R2 is selected from sulphamoyl and a group R4-E-; 
q is 0-2; wherein the values of R2 maybe the same or different; and wherein p+q=1-5; 
R3 is selected from C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C3-8cycloalkyl and heterocyclyl; wherein R3 may be optionally substituted by one or more R5; and wherein if said heterocyclyl contains an —NH— moiety that nitrogen may be optionally substituted by a group selected from R6; and wherein if said heterocyclic group contains an —NH— moiety that nitrogen may be optionally substituted by a group selected from R7; 
R4 is selected from C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C3-8cycloalkyl, C3-8cycloalkylC1-6alkyl, phenyl, a heterocyclic group phenylC1-6alkyl and (heterocyclic group)C1-6alkyl; wherein R4 may be optionally substituted on carbon by one or more R8; and wherein if said heterocyclic group contains an —NH— moiety that nitrogen may be optionally substituted by a group selected from R9; 
E is selected from —C(O)—, —N(Ra)C(O)C—, —C(O)N(Ra)—, —S(O)r—, —SO2N(Ra)— and —N(Ra)SO2—; wherein Ra is hydrogen or C1-6alkyl optionally substituted by one or more R10 and r is 1-2; 
R5 is selected from halo, nitro, cyano, hydroxy, trifluoromethoxy, amino, carboxy, carbamoyl, mercapto, sulphamoyl, C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C1-6alkoxy, C1-6alkanoyl, C1-6alkanoyloxy, N-(C1-6alkyl)amino, N,N-(C1-6alkyl)2amino, C1-6alkanoylamino, N-(C1-6alkyl)carbamoyl, N,N-(C1-6alkyl)2carbamoyl, C1-6alkylS(O)a wherein a is 0 to 2, C1-6carboxycarbonyl, N-(C1-6alkyl)sulphamoyl, N,N-(C1-6alkyl)2sulphamoyl, C3-8cycloalkyl, phenyl and a heterocyclic group; wherein R5 may be optionally substituted on carbon by one or more R11; and wherein if said heterocyclic group contains an —NH— moiety that nitrogen may be optionally substituted by a group selected from R12; 
R6, R7, R9 and R12 are independently selected from C1-4alkyl, C1-4alkanoyl, C1-4alkylsulphonyl, C1-4alkoxycarbonyl, carbamoyl, N-(C1-4alkyl)carbamoyl, N,N-(C1-4alkyl)carbamoyl, benzyl, benzyloxycarbonyl, benzoyl and phenylsulphonyl; wherein R6, R7, R9 and R12 may be independently optionally substituted on carbon by one or more R13; 
R8 and R10 are independently selected from halo, nitro, cyano, hydroxy, trifluoromethyl, trifluoromethoxy, amino, carboxy, carbamoyl, mercapto, sulphamoyl, C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C1-6alkoxy, C1-6alkanoyl, C1-6alkanoyloxy, N-(C1-6alkyl)amino, N,N-(C1-6alkyl)2amino, C1-6alkanoylamino, N-(C1-6alkyl)carbamoyl, N,N-(C1-6alkyl)2carbamoyl, C1-6alkylS(O)a wherein a is 0 to 2, C1-6alkoxycarbonyl, N-(C1-6alkyl)sulphamoyl and N,N-(C1-6alkyl)2sulphamoyl; wherein R8 and R10 may be independently optionally substituted on carbon by one or more R14; and 
R11, R13 and R14 are independently selected from halo, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino, carboxy, carbamoyl, mercapto, sulphamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino, ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulphinyl, ethylsulphinyl, mesyl, ethylsulphonyl, methoxycarbonyl, ethoxycarbonyl, N-methylsulphamoyl, N-ethylsulphonyl, N,N-dimethylsulphamoyl, N,N-diethylsulphamoyl or N-methyl-N-ethylsulphamoyl; 

or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

2. A compound of formula (I) as claimed in claim 1, wherein X is —O— or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

3. A compound of formula (I) as claimed in claim 1, wherein X is —S— or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

4. A compound of formula (I) as claimed in claim 1 wherein R1 is halo or C1-2alkyl or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

5. A compound of formula (I) as claimed in claim 1 wherein p is 0-2; wherein the values of R1 may be the same or different or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

6. A compound of formula (I) as claimed in claim 1 wherein:
R2 is sulphamoyl or a group R4-E-; wherein: 
R4 is selected from C1-6alkyl, C2-6alkenyl, C3-8cycloalkyl, C3-8cycloalkylC1-6alkyl, and (heterocyclic group)C1-6alkyl; wherein R4 may be optionally substituted on carbon by one or more R8; 
E is selected from —S(O)r— and —N(Ra)SO2—; wherein Ra is hydrogen or C1-6alkyl and r is 2; and 
R8 is selected from halo, C1-6alkoxy and N-(C1-6alkyl)amino; 

or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

7. A compound of formula (I) as claimed in claim 1 wherein q is 1 or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

8. A compound of formula (I) as claimed in claim 1 wherein R3 is selected from C1-6alkyl, C2-6alkyl and heterocyclyl; wherein R3 may be optionally substituted by one or more R5; wherein:
R5 is selected from C1-6alkoxy, C3-8cycloalkyl and a heterocyclic group; wherein R5 may be optionally substituted on carbon by one or more R11; and 
R11 is methoxy; 

or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

9. A compound of formula (I) as depicted in claim 1 wherein:
X is —O— or —S—; 
p is 0; 
R2 is sulphamoyl or a group R4-E-; wherein: 
R4 is selected from C1-6alkyl, C2-6alkenyl, C3-8cycloalkyl, C3-8cycloalkylC1-6alkyl, and (heterocyclic group)C1-6alkyl; wherein R4 may be optionally substituted on carbon by one or more R8; 
E is selected from —S(O)r— and —N(Ra)SO2—; wherein Ra is hydrogen or C1-6alkyl and r is 2; 
R8 is selected from halo, C1-6alkoxy and N-(C1-6alkyl)amino; 
q is 1; 
R3 is selected from C1-6alkyl, C2-6alkynyl and heterocyclyl; wherein R3 may be optionally substituted by one or more R5; wherein: 
R5 is selected from C1-6alkoxy, C3-8cycloalkyl and a heterocyclic group; wherein R5 may be optionally substituted on carbon by one or more R11; and 
R11 is methoxy; 

or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof.

10. A process for preparing a compound of formula (I), as claimed in claim 1, or a pharmaceutically acceptable salt or an in vivo hydrolysable ester thereof which process (wherein R1, R2, R3, p and q are, unless otherwise specified, as defined in claim 1 comprises of:
a) reaction of a pyrimidine of formula (II): 





wherein L is a displaceable group; with an aniline of formula (III): 



b) reacting a pyrimidine of formula (IV): 





wherein L is a displaceable group; with an compound of formula (V):


R3XH  (V) 


or
c) for compounds of formula (I) where R2 is sulphamoyl or a group R4-E- and E is —NHSO2—; reacting a pyrimidine of formula (VI): 





wherein Z is a displaceable group; with an amine of formula (VII):


R4—NH2  (VII) 

d) by converting a compound of formula (VIII): 





wherein Rb is hydrogen or t-butyl; into a compound of formula (I);
and thereafter, optionally:
i) converting a compound of the formula (I) into another compound of the formula (I); 
ii) removing any protecting groups; 
iii) forming a pharmaceutically acceptable salt or in vivo hydrolysable ester.

11. A pharmaceutical composition which comprises a pyrimidine derivative of the formula (I), or a pharmaceutically acceptable salt or in vivo hydrolysable ester thereof, as claimed in any one of claims 1-9, in association with a pharmaceutically-acceptable diluent or carrier.

12. A method for inhibiting a CDK2, CDK4 or CDK6 cell cycle kinase in a warm-blooded animal in need thereof, which comprises administering to said animal an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in any one of claims 1-9.