HK1099021B - Hydroxyalkyl substituted pyrido-7-pyrimidin-7-ones - Google Patents

Description

Hydroxyalkyl substituted pyrido-7-pyrimidin-7-ones

The present invention relates to pyridopyrimidines and derivatives thereof. In particular, the present invention provides 2, 6-disubstituted 7-oxo-pyrido [2, 3-d ] pyrimidines, methods for their manufacture, pharmaceutical formulations comprising these compounds and methods for their use.

Mitogen-activated protein kinases (MAPs) are a class of proline-directed serine/threonine kinases that activate their substrates by dual phosphorylation. The kinases are activated by a variety of signals including nutritional and osmotic stress, UV light, growth factors, endotoxins, and inflammatory cytokines. One group of MAP kinases is the p38 kinase group, which includes multiple isoforms (e.g., p38 α, p39 β, p38 γ, and p38 δ). p38 kinase is responsible for phosphorylating and activating transcription factors as well as other kinases, and is activated by physical and chemical stress, pro-inflammatory cytokines and bacterial lipopolysaccharide.

More importantly, the phosphorylation product of p38 has been shown to mediate the production of inflammatory cytokines, including TNF, IL-1, and cyclooxygenase-2. Each of these cytokines has been implicated in a number of disease states and conditions. For example, TNF- α is a cytokine produced primarily by activated monocytes and macrophages. Excessive or uncontrolled production of TNF- α has been considered to be one of the causes of the pathogenesis of rheumatoid arthritis. Recently, inhibition of TNF production has been shown to have broad applications in the treatment of inflammatory disorders, inflammatory bowel disease, multiple sclerosis and asthma.

TNF is also involved in viral infections such as HIV, influenza and herpes viruses, including herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), Cytomegalovirus (CMV), varicella-zoster virus (VZV), Epstein-Barr virus, human herpes virus-6 (HHV-6), human herpes virus-7 (HHV-7), human herpes virus-8 (HHV-8), pseudorabies and rhinotracheitis.

Similarly, IL-1 is produced by activated monocytes and macrophages and plays a role in many pathophysiological responses including rheumatoid arthritis, fever and reduction of bone resorption.

In addition, involvement of p38 has been implicated in stroke, alzheimer's disease, osteoarthritis, lung injury, septic shock, angiogenesis, dermatitis, psoriasis and atopic dermatitis. Opp. opin. ther. patents, 2000, 10 (1).

Inhibition of these cytokines by inhibition of p38 kinase would be beneficial in controlling, alleviating and slowing many of these disease states.

Certain 6-aryl-pyrido [2, 3-d ] pyrimidin-7-ones, -7-imines and 7-thiones are disclosed in WO 96/34867 as inhibitors of protein tyrosine kinases that mediate cell proliferation. Other 6-aryl-pyrido [2, 3-d ] pyrimidines and naphthyridines are also disclosed in WO 96/15128 as tyrosine kinase inhibitors. WO 98/33798 discloses 6-alkyl-pyrido [2, 3-d ] pyrimidin-7-ones as inhibitors of cyclin dependent kinases. Certain 4-amino-pyridopyrimidines are disclosed as dihydrofolate reductase inhibitors in EP 0278686 a 1.

One embodiment of the present invention provides compounds of formula I

Wherein

X¹Is O, C ═ O or S (O)_nWherein n is 0, 1 or 2;

Ar¹is aryl or heteroaryl;

R¹is alkoxyAn alkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, hydroxyalkyl or hydroxycycloalkyl group; and is

R²Is hydroxyalkyl, oxoalkyl or hydroxycycloalkyl.

Although certain substituted pyrido-7-pyrimidin-7-ones are known to have enzymatic activity in vitro anti-p 38 assays (see, e.g., US-2003-0171584-A1, which is incorporated herein by reference in its entirety), the present inventors have surprisingly and unexpectedly found that compounds of formula I have significantly higher activity in Lipopolysaccharide (LPS) -induced human whole blood cysteine production assays than previously disclosed compounds.

The compounds of formula I are protein kinase inhibitors and exhibit potent activity against p38 in vivo. They are selective against p38 kinase over cyclin-dependent kinases and tyrosine kinases. Thus, the compounds of the present invention are useful for treating diseases mediated by proinflammatory cytokines such as TNF and IL-1. Thus, another aspect of the invention provides a method of treating a p 38-mediated disease or condition, wherein a therapeutically effective amount of a compound of formula I is administered to a patient.

Unless otherwise stated, the following terms used in the specification and claims have the meanings given below.

"alkoxyalkyl" means a group of the formula R^a-O-R^b-a moiety wherein R^aIs alkyl and R^bIs alkylene as defined herein. Examples of alkoxyalkyl groups include, for example, 2-methoxyethyl, 3-methoxypropyl, 1-methyl-2-methoxyethyl, 1- (2-methoxyethyl) -3-methoxy-propyl, and 1- (2-methoxyethyl) -3-methoxypropyl.

"alkyl" means a linear saturated monovalent hydrocarbon group of 1 to 6 carbon atoms or a branched saturated monovalent hydrocarbon group of 3 to 6 carbon atoms, such as methyl, ethyl, propyl, 2-propyl, n-butyl, isobutyl, tert-butyl, pentyl, and the like.

"alkylene" means a linear saturated divalent hydrocarbon group of 1 to 6 carbon atoms, or a branched saturated divalent hydrocarbon group of 3 to 6 carbon atoms, for example, methylene, ethylene, 2, 2-dimethylethylene, propylene, 2-methylpropylene, butylene, pentylene, and the like.

"aryl" refers to a monovalent monocyclic or bicyclic aromatic hydrocarbon radical optionally substituted independently with one or more substituents, preferably 1, 2 or 3 substituents, preferably selected from alkyl, hydroxy, alkoxy, haloalkyl, haloalkoxy, halo, nitro, cyano, amino, monoalkylamino, dialkylamino, methylenedioxy, ethylenedioxy and acyl. Particularly preferred aryl substituents are halides. More specifically, the term aryl includes, but is not limited to, phenyl, chlorophenyl, fluorophenyl, difluorophenyl (e.g., 2, 4-and 2, 6-difluorophenyl), methoxyphenyl, 1-naphthyl, 2-naphthyl, and derivatives thereof.

"cycloalkyl" refers to a saturated monovalent cyclic hydrocarbon group of 3 to 7 ring carbons, such as cyclopropyl, cyclobutyl, cyclohexyl, 4-methyl-cyclohexyl, and the like. Cycloalkyl groups may be optionally substituted with one or more substituents, preferably 1, 2, or 3 substituents. Preferably, the substituents of the cycloalkyl group are selected from the group consisting of alkyl, hydroxy, alkoxy, haloalkyl, haloalkoxy, halo, amino, monoalkylamino, dialkylamino and acyl. Particularly preferred substituents for cycloalkyl groups include alkyl, hydroxy, alkoxy, haloalkyl, haloalkoxy and halo. Particularly preferred cycloalkyl substituents include alkyl, hydroxy, alkoxy and halo.

"cycloalkylalkyl" refers to the formula R^c-R^d-a moiety wherein R^cIs cycloalkyl as defined herein, R^dIs alkylene as defined herein.

"halo" and "halide" are used interchangeably herein and refer to fluorine, chlorine, bromine or iodine. Preferred halides are fluorine and chlorine, with fluorine being a particularly preferred halide.

"haloalkyl" means substituted with one or moreAlkyl radicals, substituted by the same or different halogen atoms, e.g. -CH₂Cl，-CF₃，-CH₂CF₃，-CH₂CCl₃And the like.

"heteroaryl" means a monovalent monocyclic or bicyclic radical of 5 to 12 ring atoms containing at least one aromatic ring containing 1, 2 or 3 ring heteroatoms selected from N, O or S (preferably N or O), the remaining ring atoms being C, with the understanding that the point of attachment of the heteroaryl radical is on the aromatic ring. The heteroaryl ring is optionally independently substituted with one or more substituents, preferably 1 or 2 substituents, selected from alkyl, haloalkyl, hydroxy, alkoxy, halo, nitro or cyano. More specifically, the term heteroaryl includes, but is not limited to, pyridyl, furyl, thienyl, thiazolyl, isothiazolyl, triazolyl, imidazolyl, isoxazolyl, pyrrolyl, pyrazolyl, pyrimidinyl, benzofuryl, tetrahydrobenzofuryl, isobenzofuryl, benzothiazolyl, benzisothiazolyl, benzotriazolyl, indolyl, isoindolyl, benzoxazolyl, quinolinyl, tetrahydroquinolinyl, isoquinolinyl, benzimidazolyl, benzisoxazolyl or benzothienyl, imidazo [1, 2-a ] -pyridyl, imidazo [2, 1-b ] thiazolyl, and derivatives thereof.

"Heterocyclyl" means a saturated or unsaturated non-aromatic cyclic group of 3 to 8 ring atoms, 1 or 2 of which are selected from N, O or S (O)_n(wherein N is an integer of 0 to 2), preferably a heteroatom of N or O, the remaining ring atoms being C, wherein 1 or 2 carbon atoms may optionally be substituted by carbonyl groups. The heterocyclyl ring may be optionally independently substituted with 1, 2 or 3 substituents selected from alkyl, haloalkyl, hydroxyalkyl, halo, nitro, cyano, cyanoalkyl, hydroxy, alkoxy, amino, monoalkylamino, dialkylamino, aralkyl, - (X)_n-C(O)R^e(wherein X is O or NR)^fN is 0 or 1, R^eIs hydrogen (wherein X is NR)^f) Alkyl, haloalkyl, hydroxy (when n is 0), alkoxy, amino, monoalkylamino, dialkylamino, or optionally substituted phenyl, and R^fIs H or alkyl), -alkylene-C (O) R^g(wherein R is^gIs alkyl, -OR^hOr NRⁱR^j，R^hIs hydrogen, alkyl or haloalkyl, and RⁱAnd R^jIndependently hydrogen or alkyl), or-S (O)_nR^k(wherein n is an integer of 0 to 2) such that when n is 0, R^kIs hydrogen, alkyl, cycloalkyl or cycloalkylalkyl, and when n is 1 or 2, R is^kIs alkyl, cycloalkyl, cycloalkylalkyl, amino, acylamino, monoalkylamino or dialkylamino. Particularly preferred heterocyclyl substituents include alkyl, haloalkyl, hydroxyalkyl, halo, hydroxy, alkoxy, amino, monoalkylamino, dialkylamino, aralkyl and-S (O)_nR^k. Specifically, the term heterocyclyl includes, but is not limited to, tetrahydrofuranyl, pyridinyl, tetrahydropyranyl, piperidino, N-methylpiperidin-3-yl, piperazino, N-methylpyrrolidin-3-yl, 3-pyrrolidinyl, morpholino, thiomorpholino-1-oxide, thiomorpholino-1, 1-dioxide, 4- (1, 1-dioxo-tetrahydro-2H-thiopyranyl), pyrrolinyl, imidazolinyl, N-methanesulfonyl-piperidin-4-yl, and derivatives thereof, each of which may be optionally substituted.

"hydroxyalkyl" means an alkyl moiety as defined herein substituted with one or more, preferably 1, 2 or 3, hydroxyl groups, provided that the number of hydroxyl groups carried by the same carbon atom does not exceed 1. Representative examples include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, 1- (hydroxymethyl) -2-methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl, 4-hydroxybutyl, 2, 3-dihydroxypropyl, 2-hydroxy-1-hydroxymethylethyl, 2, 3-dihydroxybutyl, 3, 4-dihydroxybutyl and 2- (hydroxymethyl) -3-hydroxypropyl.

"Hydroxycycloalkyl" refers to a cycloalkyl moiety as defined herein wherein 1, 2, or 3 hydrogen atoms in the cycloalkyl group have been substituted with a hydroxy substituent. Representative examples include, but are not limited to, 2-, 3-, or 4-hydroxycyclohexyl and the like.

"leaving group" has the meaning commonly associated therewith in synthetic organic chemistry, i.e., an atom or group capable of being substituted with a nucleophile, including halogen (e.g., chlorine, bromine, and iodine), alkylsulfonyloxy, arylsulfonyloxy, alkylcarbonyloxy (e.g., acetoxy), arylcarbonyloxy, methylsulfonyloxy, tosyloxy, trifluoromethanesulfonyloxy, aryloxy (e.g., 2, 4-dinitrophenoxy), methoxy, N, O-dimethylhydroxyamino, and the like.

"optionally substituted phenyl" refers to a phenyl ring optionally substituted independently with one or more substituents, preferably 1 or 2 substituents, selected from alkyl, hydroxy, alkoxy, haloalkyl, haloalkoxy, halo, nitro, cyano, amino, methylenedioxy, ethylenedioxy, and acyl.

"oxoalkyl" refers to an alkyl group substituted with one or more carbonyloxy moieties (i.e., ═ O), for example, of the formula R^z-C(＝O)-R^yA moiety of (a) wherein R^yIs alkylene, R^zIs an alkyl group. Representative oxoalkyl groups include 2-propanon-3-yl, 2-methyl-3-butanone-4-yl, and the like.

"pharmaceutically acceptable excipient" means an excipient that can be used in the preparation of pharmaceutical compositions, which are generally safe, non-toxic, neither biologically nor otherwise undesirable. The term includes excipients that are useful for veterinary and human pharmaceutical use. As used in the specification and claims, the term "pharmaceutically acceptable excipient" includes one or more such excipients.

"pharmaceutically acceptable salts" of a compound refer to salts that are pharmaceutically acceptable and possess the desired pharmacological activity of the parent compound. Such salts include: (1) acid addition salts with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1, 2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2.2.2] -oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, dodecylsulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, and the like; or (2) a salt formed when an acidic proton present in the parent compound is replaced with a metal ion, such as an alkali metal ion, an alkaline earth metal ion, or an aluminum ion; or salts formed when coordinated with organic bases such as ethanolamine, diethanolamine, triethanolamine, trimethylamine (tromethamine), N-methylglucamine.

The terms "prodrug" and "prodrug" are used interchangeably herein and refer to any compound that releases an active parent drug according to formula I in vivo when such prodrug is administered to a mammalian subject. The prodrugs of the compounds of formula I are prepared by: by modifying one or more functional groups present in the compounds of formula I, the modification may be cleaved in vivo to release the parent compound. Prodrugs include compounds of formula I wherein a hydroxy, amino, sulfhydryl, carboxy or carbonyl group in the compound of formula I is bonded to any group that can be cleaved in vivo to yield the free hydroxy, amino or sulfhydryl group, respectively. Examples of Prodrugs include, but are not limited to, esters (e.g., acetate, dialkylaminoacetate, formate, phosphate, sulfate, and benzoate derivatives) and carbamates (e.g., N-dimethylaminocarbonyl) of hydroxyl functional groups in the compounds of formula I, ester groups (e.g., ethyl esters, morpholino ethanol esters) of carboxyl functional groups, N-acyl derivatives (e.g., N-acetyl) of amino functional groups, N-Mannich bases, Schiff bases and enaminones, oximes, acetals, ketals and enol esters of ketone and aldehyde functional groups, and the like, see Bundegard, "Design of produgs" p1-92, Elsevier, New York-Oxford (1985).

"protecting group" refers to a group of atoms that, when attached to a reactive group in a molecule, masks, reduces, or prevents the reactivity of the attached group. Examples of protecting Groups can be found in Green and Wuts, Protective Groups in Organic Chemistry (Wiley, second edition 1991), and Harrison et al, Complex of Synthetic Organic Methods, Vol.1-8 (John Wiley and Sons, 1971-. Representative amino protecting groups include formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl (CBZ), tert-butoxycarbonyl (Boc), Trimethylsilyl (TMS), 2-trimethylsilyl-ethanesulfonyl (SES), trityl and substituted trityl, allyloxycarbonyl, 9-Fluorenylmethoxycarbonyl (FMOC), nitro-veratryloxycarbonyl (NVOC), and the like. Representative hydroxy protecting groups include groups which acylate or alkylate the hydroxy group, such as benzyl and trityl ethers as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers and allyl ethers.

"treatment" or "therapy" of a disease includes: (1) preventing a disease, i.e., such that the clinical symptoms of the disease do not develop in a mammal that may be exposed to or susceptible to the disease, but does not yet experience or manifest the symptoms of the disease; (2) inhibiting a disease, i.e., arresting or reducing the development of the disease or its clinical symptoms; and (3) relief of the disease, i.e., causing regression of the disease or its clinical symptoms.

"therapeutically effective amount" means the amount of a compound that, when administered to a mammal to treat a disease, is sufficient to effect treatment for the disease. The "therapeutically effective amount" will vary depending on the compound, the disease and its severity, and the age, weight, etc., of the mammal being treated.

The terms "treating", "contacting" or "reacting", when referring to a chemical reaction, refer to the addition or mixing of two or more reagents under suitable conditions to produce an indicated and/or desired product. It will be appreciated that the reaction that produces the indicated and/or the desired product may not necessarily result directly from the combination of the two reagents that were initially added, i.e., one or more intermediates may be produced in the mixture that ultimately results in the production of the indicated and/or the desired product.

The compounds of the invention may exist in unsolvated or solvated forms, including hydrated forms. In general, solvated forms, including hydrated forms, and unsolvated forms are equivalent and are intended to be encompassed within the scope of the present invention. In addition to the compounds described above, the compounds of the present invention include all tautomeric forms. Furthermore, the invention also includes all pharmaceutically acceptable salt forms of these compounds as well as all stereoisomers of the pro-drug forms and the pure chiral forms or the racemic or other mixture forms of said compounds.

The compounds of formula I are also capable of forming pharmaceutically acceptable acid addition salts. All of these forms are within the scope of the present invention.

Pharmaceutically acceptable acid addition salts of the compounds of formula I include salts derived from inorganic acids such as hydrochloric, nitric, phosphoric, sulfuric, hydrobromic, hydroiodic, phosphorous and the like, as well as salts derived from organic acids such as aliphatic mono-and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids and the like. Thus, such salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, caprylate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartrate, methanesulfonate, and the like. Also contemplated are salts of amino acids, such as arginine salts and the like, and gluconates, galacturonates (see, e.g., Berge et al, J.of pharmaceutical Science 66: 1-19 (1977)).

Acid addition salts of basic compounds can be prepared by the following method: the free base form is contacted with a sufficient amount of the desired acid to form a salt in a conventional manner. The free base form is again produced by contacting the salt form with a base and isolating the free base in a conventional manner. The free base forms differ from their corresponding salt forms in certain physical properties, such as solubility in polar solvents, but for the purposes of the present invention, these salts are equivalent to their corresponding free bases.

In one embodiment, Ar¹Is an aryl group. Particularly preferred Ar¹Is optionally substituted phenyl. In certain embodiments, Ar¹Is phenyl optionally substituted one or more times by alkyl, halo, haloalkyl or alkoxy. More preferred Ar¹Is a disubstituted phenyl group, for example a 2, 4-disubstituted phenyl group. Still more preferably Ar¹Is a 2, 4-dihalo-substituted phenyl group. Particularly preferred Ar¹Is 2, 4-difluorophenyl.

In another embodiment, X¹Is O.

In another embodiment, R¹Is alkoxyalkyl, alkyl, cycloalkyl, cycloalkylalkyl, hydroxyalkyl or heterocyclyl. Within this group, R is particularly preferred¹Including optionally substituted tetrahydropyranyl, 1-methyl-2-methoxyethyl, optionally substituted cyclopentyl, optionally substituted cyclopropyl, isopropyl, optionally substituted cyclohexyl, 1- (2-hydroxyethyl) -3-hydroxypropyl, 1-hydroxymethyl-2-hydroxypropyl, 1-hydroxymethyl-3-hydroxypropyl, 1-methylpropyl, 2-hydroxy-1-methylethyl, 1- (2-methoxyethyl) -3-methoxypropyl, N-methanesulfonyl piperidinyl, ethyl, methyl, 2-hydroxypropyl, neopentyl, 1-dimethyl-2-hydroxyethyl, 1- (hydroxymethyl) propyl, 2-methylpropyl, cyclopropylmethyl, optionally substituted cyclobutyl, 1, 2-dimethyl-2-hydroxypropyl and 1- (hydroxymethyl) -2-hydroxyethyl.

In yet another embodiment, R is preferred¹Is hydroxyalkyl, with 2-hydroxy-1-methylethyl being particularly preferred R¹. Especially preferred R¹Comprising enantiomerically enriched 2-hydroxy-1-methylethyl, i.e.(R) -and (S) -2-hydroxy-1-methylethyl.

In a specific embodiment, R²Is 2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropyl, 1- (2-hydroxyethyl) -3-hydroxypropyl or 2-oxopropyl.

In another embodiment, R²Is hydroxyalkyl. Within this group, R is particularly preferred²Are 2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropyl and 1- (2-hydroxyethyl) -3-hydroxypropyl. Especially preferred R²Is 2-hydroxypropyl. In another embodiment, R²Is an oxoalkyl group.

Still further, combinations of the preferred groups described herein form other preferred embodiments. For example, in a particularly preferred embodiment, R¹Is (R) -or (S) -2-hydroxy-1-methylethyl, R²Is (R) -or (S) -2-hydroxypropyl, or 2-oxopropyl, X¹Is O, and Ar¹Is 2, 4-difluorophenyl.

An embodiment of the invention provides compounds of formula I, wherein Ar¹Is aryl, and X¹Is O. In another embodiment of the invention, there is provided a compound of formula I, wherein Ar is¹Is aryl, X¹Is O, and R¹Is alkoxyalkyl, alkyl, cycloalkyl, cycloalkylalkyl, hydroxyalkyl or heterocyclyl. In yet another embodiment of the present invention, there is provided a compound of formula I, wherein Ar is¹Is aryl, X¹Is O, and R¹Is tetrahydropyranyl, 1-methyl-2-methoxyethyl, cyclopentyl, cyclopropyl, isopropyl, cyclohexyl, 1- (2-hydroxyethyl) -3-hydroxypropyl, 1-hydroxymethyl-2-hydroxypropyl, 1-hydroxymethyl-3-hydroxypropyl, 1-methylpropyl, 2-hydroxy-1-methylethyl, 1- (2-methoxyethyl) -3-methoxypropyl, N-methanesulfonylpiperidinyl, ethyl, methyl, 2-hydroxypropyl, neopentyl, 1, 1-dimethyl-2-hydroxyethyl, 1- (hydroxymethyl) propyl, 2-methylpropyl, cyclopropylmethyl, cyclobutyl, 1, 2-dimethyl-2-hydroxypropyl or 1- (hydroxymethyl) -2-hydroxyethyl. In yet another embodiment of the present inventionFor compounds of formula I, wherein Ar¹Is aryl, X¹Is O, R¹Is tetrahydropyranyl, 1-methyl-2-methoxyethyl, cyclopentyl, cyclopropyl, isopropyl, cyclohexyl, 1- (2-hydroxyethyl) -3-hydroxypropyl, 1-hydroxymethyl-2-hydroxypropyl, 1-hydroxymethyl-3-hydroxypropyl, 1-methylpropyl, 2-hydroxy-1-methylethyl, 1- (2-methoxyethyl) -3-methoxypropyl, N-methanesulfonylpiperidinyl, ethyl, methyl, 2-hydroxypropyl, neopentyl, 1, 1-dimethyl-2-hydroxyethyl, 1- (hydroxymethyl) propyl, 2-methylpropyl, cyclopropylmethyl, cyclobutyl, 1, 2-dimethyl-2-hydroxypropyl, or 1- (hydroxymethyl) -2-hydroxyethyl, and R²Is 2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropyl, 1- (2-hydroxyethyl) -3-hydroxypropyl, or 2-oxopropyl.

An embodiment of the invention provides compounds of formula I, wherein Ar¹Is aryl, X¹Is O, R¹Is (R) -2-hydroxy-1-methylethyl or (S) -2-hydroxy-1-methylethyl, and R²Is 2-oxopropyl, (R) -2-hydroxypropyl or (S) -2-hydroxypropyl.

An embodiment of the invention provides compounds of formula I, wherein Ar¹Is aryl, X¹Is O and R¹Is hydroxyalkyl. Another embodiment of the present invention provides compounds of formula I wherein Ar¹Is aryl, X¹Is O, R¹Is hydroxyalkyl, and R²Is hydroxyalkyl. In a further embodiment of the invention there are provided compounds of formula I wherein Ar¹Is aryl, X¹Is O, R¹Is hydroxyalkyl, and R²Is 2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropyl or 1- (2-hydroxyethyl) -3-hydroxypropyl.

Another embodiment of the present invention provides compounds of formula I wherein Ar¹Is aryl, X¹Is a group of compounds represented by the general formula (I),

R¹is (R) -2-hydroxy-1-methylethyl and R²Is (R) -2-hydroxypropyl;

R¹is (R) -2-hydroxy-1-methylethylRadical and R²Is (S) -2-hydroxypropyl;

R¹is (S) -2-hydroxy-1-methylethyl and R²Is (R) -2-hydroxypropyl; or

R¹Is (S) -2-hydroxy-1-methylethyl and R²Is (S) -2-hydroxypropyl.

An embodiment of the invention provides compounds of formula I, wherein R¹Is hydroxyalkyl. Another embodiment of the invention provides compounds of formula I wherein R¹Is hydroxyalkyl and R²Is hydroxyalkyl. In a further embodiment the invention provides a compound of formula I wherein R¹Is hydroxyalkyl, R²Is hydroxyalkyl and Ar¹Is an aryl group.

An embodiment of the invention provides compounds of formula I, wherein R²Is hydroxyalkyl.

In certain embodiments, the compound of the invention may be a compound of formula II

Wherein

m is 0 to 4;

each R³Is alkyl, halogen, alkoxy or haloalkyl; and is

R¹And R²As defined herein.

In a particular embodiment, m is 1 and R³Is a halogen.

In another embodiment, m is 2 and R³Is a halogen.

An embodiment of the invention provides a compound of formula II, wherein R¹Is alkoxyalkyl, alkyl, cycloalkyl, cycloalkylalkyl, hydroxyalkylAlkyl, or heterocyclyl.

An embodiment of the invention provides a compound of formula II, wherein R¹Is tetrahydropyranyl, 1-methyl-2-methoxyethyl, cyclopentyl, cyclopropyl, isopropyl, cyclohexyl, 1- (2-hydroxyethyl) -3-hydroxypropyl, 1-hydroxymethyl-2-hydroxypropyl, 1-hydroxymethyl-3-hydroxypropyl, 1-methylpropyl, 2-hydroxy-1-methylethyl, 1- (2-methoxyethyl) -3-methoxypropyl, N-methanesulfonylpiperidinyl, ethyl, methyl, 2-hydroxypropyl, neopentyl, 1, 1-dimethyl-2-hydroxyethyl, 1- (hydroxymethyl) propyl, 2-methylpropyl, cyclopropylmethyl, cyclobutyl, 1, 2-dimethyl-2-hydroxypropyl or 1- (hydroxymethyl) -2-hydroxyethyl. Another embodiment of the present invention provides compounds of formula II wherein R¹Is tetrahydropyranyl, 1-methyl-2-methoxyethyl, cyclopentyl, cyclopropyl, isopropyl, cyclohexyl, 1- (2-hydroxyethyl) -3-hydroxypropyl, 1-hydroxymethyl-2-hydroxypropyl, 1-hydroxymethyl-3-hydroxypropyl, 1-methylpropyl, 2-hydroxy-1-methylethyl, 1- (2-methoxyethyl) -3-methoxypropyl, N-methanesulfonylpiperidinyl, ethyl, methyl, 2-hydroxypropyl, neopentyl, 1, 1-dimethyl-2-hydroxyethyl, 1- (hydroxymethyl) propyl, 2-methylpropyl, cyclopropylmethyl, cyclobutyl, 1, 2-dimethyl-2-hydroxypropyl, or 1- (hydroxymethyl) -2-hydroxyethyl, and R²Is 2-hydroxyethyl, 3-hydroxypropyl, 2-hydroxypropyl, 1- (2-hydroxyethyl) -3-hydroxypropyl or 2-oxopropyl.

An embodiment of the invention provides a compound of formula II, wherein R¹Is (R) -2-hydroxy-1-methylethyl or (S) -2-hydroxy-1-methylethyl, and R²Is 2-oxopropyl, (R) -2-hydroxypropyl or (S) -2-hydroxypropyl.

An embodiment of the invention provides a compound of formula II, wherein R¹And R²Are all hydroxyalkyl.

An embodiment of the present invention provides compounds of formula II, wherein

R¹Is (R) -2-hydroxy-1-methylethyl and R²Is (R) -2-hydroxypropyl;

R¹is (R) -2-hydroxy-1-methylethyl and R²Is (S) -2-hydroxypropyl;

R¹is (S) -2-hydroxy-1-methylethyl and R²Is (R) -2-hydroxypropyl; or

R¹Is (S) -2-hydroxy-1-methylethyl and R²Is (S) -2-hydroxypropyl.

Another embodiment of the present invention provides compounds of formula II wherein R¹And R²Is hydroxyalkyl, n is 1 and R³Is a halo group.

Another embodiment of the present invention provides compounds of formula II wherein R¹And R²Is hydroxyalkyl, n is 2 and R³Is a halo group.

Representative compounds according to the invention are shown in table 1.

TABLE 1

#	Name (R)
		1	6- (2, 4-difluoro-phenoxy) -8- (2-hydroxy-ethyl) -2- (tetrahydropyran-4-ylamino) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
2	6- (2, 4-difluoro-phenoxy) -8- (3-hydroxy-propyl) -2- (tetrahydropyran-4-ylamino) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		3	6- (2, 4-difluoro-phenoxy) -8- (2-hydroxy-propyl) -2- (tetrahydropyran-4-ylamino) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
4	6- (2, 4-difluoro-phenoxy) -8- [ 3-hydroxy-1- (2-hydroxy-ethyl) -propyl]-2- ((S) -2-methoxy-1-methyl-ethylamino) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		5	2-cyclopentylamino-6- (2, 4-difluoro-phenoxy) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
6	2-cyclopropylamino-6- (2, 4-difluoro-phenoxy) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		7	6- (2, 4-difluoro-phenoxy) -8- ((S) -2-hydroxy-propyl) -2-isopropylamino-8H-pyrido [2, 3-d]Pyrimidin-7-ones
8	2-cyclohexylamino-6- (2, 4-difluoro-phenoxy) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		9	6- (2, 4-difluoro-phenoxy) -2- [ 3-hydroxy-1- (2-hydroxy-ethyl) -propylamino]-8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
10	6- (2, 4-difluoro-phenoxy) -8- ((S) -2-hydroxy-propaneYl) -2- (tetrahydropyran-4-ylamino) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		11	6- (2, 4-difluoro-phenoxy) -2- ((1R, 2R) -2-hydroxy-1-hydroxymethyl-propylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
12	6- (2, 4-difluoro-phenoxy) -2- ((S) -3-hydroxy-1-hydroxymethyl-propylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		13	6- (2, 4-difluoro-phenoxy) -2- ((R) -3-hydroxy-1-hydroxymethyl-propylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones

14	2- ((S) -sec-butylamino) -6- (2, 4-difluoro-phenoxy) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		15	2- ((R) -sec-butylamino) -6- (2, 4-difluoro-phenoxy) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
16	6- (2, 4-difluoro)-phenoxy) -2- ((S) -2-hydroxy-1-methyl-ethylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		17	6- (2, 4-difluoro-phenoxy) -2- ((R) -2-hydroxy-1-methyl-ethylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-f]Pyrimidin-7-ones
18	6- (2, 4-difluoro-phenoxy) -8- ((S) -2-hydroxy-propyl) -2- ((S) -2-methoxy-1-methyl-ethylamino) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		19	6- (2, 4-difluoro-phenoxy) -8- ((S) -2-hydroxy-propyl) -2- [ 3-methoxy-1- (2-methoxy-ethyl) -propylamino]-8H-pyrido [2, 3-d]Pyrimidin-7-ones
20	6- (2, 4-difluoro-phenoxy) -8- ((R) -2-hydroxy-propyl) -2- (tetrahydropyran-4-ylamino) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		21	6- (2, 4-difluoro-phenoxy) -8- ((S) -2-hydroxy-propyl) -2- (1-methanesulfonyl-piperidin-4-ylamino) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
22	2-cyclopropylamino-6- (2, 4-difluoro-phenoxy) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		23	6- (2, 4-difluoro-phenoxy) -8- ((R) -2-hydroxy-propyl) -2-isopropylamino-8H-pyrido [2, 3-d]Pyrimidin-7-ones
24	6- (2, 4-difluoro-phenoxy) -2-ethylamino-8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		25	6- (2, 4-difluoro-phenoxy) -8- ((R) -2-hydroxy-propyl) -2-methylamino-8H-pyrido [2, 3-d]Pyrimidin-7-ones
26	2- ((S) -sec-butylamino) -6- (2, 4-difluoro-phenoxy) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		27	2- ((R) -sec-butylamino) -6- (2, 4-difluoro-phenoxy) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
28	6- (2, 4-difluoro-phenoxy) -8- ((S) -2-hydroxy-propyl) -2- ((R) -2-hydroxy-propylamino) -8H-pyrido [2, 3-d]Pyrimidin-7-ones

29	6- (2, 4-difluoro-phenoxy) -8- ((S) -2-hydroxy-propyl) -2- ((S) -2-hydroxy-propylamino) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		30	6- (2, 4-difluoro-phenoxy) -2- (2, 2-dimethyl-propylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
31	6- (2, 4-difluoro-phenoxy) -2- (2-hydroxy-1, 1-dimethyl-ethylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		32	6- (2, 4-difluoro-phenoxy) -2- ((S) -1-hydroxymethyl-propylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
33	6- (2, 4-difluoro-phenoxy) -2- ((R) -1-hydroxymethyl-propylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		34	6- (2, 4-difluoro-phenoxy) -8- ((R) -2-hydroxy-propyl) -2- ((S) -2-hydroxy-propylamino) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
35	6- (2, 4-difluoro-phenoxy) -8- ((R) -2-hydroxy-propyl) -2- ((R) -2-hydroxy-propylamino) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		36	6- (2, 4-difluoro-phenoxy) -2- (2-hydroxy-1, 1-dimethyl-ethylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
37	6- (2, 4-difluoro-phenoxy)) -2- ((R) -1-hydroxymethyl-propylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		38	6- (2, 4-difluoro-phenoxy) -8- ((R) -2-hydroxy-propyl) -2-isobutylamino-8H-pyrido [2, 3-d]Pyrimidin-7-ones
39	2- (cyclopropylmethyl-amino) -6- (2, 4-difluoro-phenoxy) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		40	2-Cyclobutylamino-6- (2, 4-difluoro-phenoxy) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
41	6- (2, 4-difluoro-phenoxy) -2- ((S) -2-hydroxy-1-methyl-ethylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		42	6- (2, 4-difluoro-phenoxy) -2- ((R) -2-hydroxy-1-methyl-ethylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
43	6- (2, 4-difluoro-phenoxy) -2- (2, 2-dimethyl-propylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones

44	6- (2, 4-difluoro-phenoxy) -2- ((S) -2-hydroxy-1, 2-dimethyl-propylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		45	6- (2, 4-difluoro-phenoxy) -2- ((S) -2-hydroxy-1-methyl-ethylamino) -8- (2-oxo-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
46	6- (2, 4-difluoro-phenoxy) -2- ((R) -2-hydroxy-1-methyl-ethylamino) -8- (2-oxo-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		47	6- (2, 4-difluoro-phenoxy) -2- (2-hydroxy-1-hydroxymethyl-ethylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
48	6- (2, 4-difluoro-phenoxy) -2- (R) -2-hydroxy-1-methyl-ethylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones
		49	6- (2, 4-Difluorophenoxy) -2- ((R) -2-hydroxy-1-methyl-ethylamino) -8- (2-oxopropyl) -8H-pyrido [2, 3-d]Pyrimidin-7-ones

While forms of the invention herein are presently considered to constitute preferred embodiments, many others are possible. It is not intended that the specification mention all of these possible equivalents or ramifications of the invention, and it is to be understood that the terms used herein are merely descriptive rather than limiting and that various changes may be made without departing from the spirit and scope of the invention.

The compounds of the present invention may be prepared by a variety of methods, including those disclosed in the well-known US-2003-0171584-A1, previously incorporated by reference. In one aspect of the invention, a process for preparing compounds of formula I is shown in scheme 1 below. It will be appreciated that although the schemes often show precise structures, the methods of the invention are broadly applicable to analogous compounds of formula I, with due consideration for the protection and deprotection of reactive functional groups by standard methods in the field of organic chemistry. For example, during chemical reactions at other sites in the molecule, it is sometimes necessary to protect hydroxyl groups (e.g., to convert to ethers or esters) in order to prevent undesirable side reactions. The hydroxyl protecting group is then removed to provide the free hydroxyl group. Similarly, amino groups and carboxylic acid groups can be protected (e.g., by derivatization) to prevent their undesirable side reactions. Typical protecting Groups and methods for attaching and cleaving them are well described in the above-incorporated references, such as Greene and Wuts, Protective Groups in Organic Synthesis, third edition, John Wiley & Sons, New York, 1999, and Harrison et al, Compendium of Synthetic Organic methods, Vols.1-8(John Wiley and Sons, 1971-1996).

Scheme 1

With hydroxyalkyl amines (R)²-NH₂) Treating the compound of formula Ia to obtain the compound of formula Ib. The reaction is conveniently carried out in a solvent which is inert under the reaction conditions, preferably a halogenated aliphatic hydrocarbon, especially dichloromethane, an optionally halogenated aromatic hydrocarbon, orA chain or cyclic ether such as Tetrahydrofuran (THF), formamide or a lower alkanol. Suitably, the reaction is carried out at a temperature of from about-20 ℃ to about 120 ℃, typically at about 0 ℃. A base, such as a trialkylamine, preferably triethylamine, is often added to the reaction mixture.

Reduction of the compound of formula Ib provides the alcohol of formula Ic. This reduction is typically carried out using lithium aluminum hydride in a manner well known to those skilled in the art (e.g., in a solvent which is inert under the reaction conditions, preferably an open chain or cyclic ether, especially THF, at from about-20 ℃ to about 70 ℃, preferably from about 0 ℃ to about Room Temperature (RT)).

Oxidation of the alcohol of formula Ic affords the aldehyde of formula Id. The oxidation is typically carried out with manganese dioxide, although a variety of other methods can also be employed (see, e.g., A)_DVANCED O_RGANIC C_HEMISTRY，4^TH _ED.，March，John Wiley &Sons, New York (1992)). Depending on the oxidizing agent employed, the reaction is conveniently carried out in a solvent which is inert under the particular oxidation conditions, preferably a halogenated aliphatic hydrocarbon, in particular dichloromethane, or an optionally halogenated aromatic hydrocarbon. The oxidation is suitably carried out at a temperature of from about 0 ℃ to about 60 ℃.

Aldehydes and esters Ar of the formula Id¹-X¹CH₂-CO₂R '(wherein R' is alkyl, Ar)¹And X¹As defined herein) in the presence of a base to give a compound of formula Ie. Any relatively non-nucleophilic base may be used, including carbonates, such as potassium carbonate, lithium carbonate and sodium carbonate; bicarbonates, such as potassium bicarbonate, lithium bicarbonate and sodium bicarbonate; amines, such as secondary and tertiary amines; and resin-bound amines, e.g. 1, 3, 4, 6, 7, 8-hexahydro-2H-pyrimido [1, 2-a ]]A pyrimidine. The reaction is conveniently carried out in a relatively polar solvent but inert under the reaction conditions, preferably in an amide such as dimethylformamide, an N-substituted pyrrolidone, especially 1-methyl-2-pyrrolidone, and at a temperature of from about 25 ℃ to about 150 ℃.

With the use of an oxidizing agent,for example, peracids such as 3-chloroperbenzoic acid (i.e., MCPBA) orOxidation of Ie gives the sulfone (If), which can be converted into a variety of target compounds. Typically, the oxidation of Ie is carried out in a solvent which is inert under the oxidation conditions. For example, when MCPBA is used as the oxidizing agent, the preferred solvent is a halogenated aliphatic hydrocarbon, particularly chloroform. When in useWhen the oxidizing agent is used, the solvent is preferably methanol, an aqueous ethanol solution or an aqueous THF solution. The reaction temperature depends on the solvent used. For organic solvents, the reaction temperature is generally from about-20 ℃ to about 50 ℃, preferably from about 0 ℃ to about RT. When water is used as solvent, the reaction temperature is generally from about 0 ℃ to about 50 ℃, preferably from about 0 ℃ to about RT. Alternatively, the oxidation may be carried out under catalytic conditions using rhenium/peroxide based reagents, see (Lahti et al, Inorg. chem., 2000, 39, 2164-.

If compound and amine (R)¹-NH₂) The reaction gives the compound of formula I. The reaction may be carried out in the presence or absence of a solvent. Conveniently, the reaction is carried out at a temperature of from about 0 ℃ to about 200 ℃, more preferably at a temperature of from about RT to about 150 ℃. Alternatively, in certain cases where sulfone If is not used, thioether Ie or the corresponding sulfoxide can be reacted with an amine (R)¹-NH₂) Directly reacting to obtain the compound shown in the formula I.

Those skilled in the art will appreciate that certain modifications of the above described arrangements are contemplated and are within the scope of the present invention. For example, certain steps will involve the use of protecting groups for functional groups that are incompatible with the particular reaction conditions.

Alternatively, the compounds of formula I may also be prepared by the method shown in scheme 2 below. Although the reactions of scheme 2 are shown as specific compounds, it will be apparent to those skilled in the art that the method of scheme 2 can be used for all compounds of the present invention.

Treatment of diethyl acetal IIa with thiourea affords the pyrimidine compound IIb, as shown in scheme 2. The reaction is conveniently carried out in an alcoholic solvent in the presence of a base such as sodium methoxide. The thiol group is then methylated, for example, with methyl iodide, to give thioether IIc.

The thioether IIc can then be treated with an α -aryloxyester IId, such as ethyl (2, 4-difluorophenoxy) acetate, to provide a pyrido-pyrimidone thioether IIe. The reaction may be carried out, for example, by heating in n-methylpyrrolidone or other polar aprotic solvents in the presence of sodium carbonate or other weak bases.

The thioether IIe reacts with carbonic ester such as propylene carbonate and the like under the condition of a polar aprotic solvent to obtain the N-hydroxyalkyl pyrido-pyrimidone thioether IIf. The reaction can be promoted by heating in the presence of potassium carbonate.

The thioether IIf is then oxidized to give the corresponding pyrido-pyrimidone sulfone IIg. The oxidation reaction may be carried out using hydrogen peroxide in a polar solvent such as dichloromethane in the presence of acetic acid. Alternatively, it can be used in the manner described in scheme 1 aboveOr MCPBA for this step of oxidation.

Treatment of sulfone IIg with a hydroxylamine, wherein the hydroxyl group is suitably protected, affords pyrido-pyrimidinone compounds IIh according to the invention. The reaction can be carried out by heating as described above with reference to scheme 1.

Scheme 2

The pyridopyrimidinone IIf can also be prepared by alkylating the pyridopyrimidinone IIe with an epoxide instead of a carbonate, as shown in scheme 3 below. The reaction of scheme 3 can be carried out by heating compound IIe under pressure in the presence of excess propylene oxide under N-methylpyrrolidone or other polar aprotic solvent conditions.

Scheme 3

The compounds of formula I may be used as medicaments, for example in the form of pharmaceutical preparations. The pharmaceutical preparations can be administered enterally, e.g. orally in the form of tablets, coated tablets, dragees, hard or soft gelatine capsules, solutions, emulsions or suspensions, nasally, e.g. in the form of nasal sprays, or rectally, e.g. in the form of suppositories. But may also be administered parenterally, for example in the form of injections.

Another aspect of the invention provides a pharmaceutical formulation comprising a compound of formula I and a pharmaceutically acceptable carrier, diluent or excipient.

Pharmaceutical formulations may be manufactured by processing a compound of formula I with a pharmaceutically inert, inorganic or organic carrier. For example, lactose, corn starch or derivatives thereof, talc, stearic acid or its salts and the like can be used as such carriers for tablets, coated tablets, dragees and hard gelatine capsules. Suitable carriers for soft gelatine capsules are, for example, vegetable oils, waxes, fats, semi-solid and liquid polyols and the like; however, depending on the nature of the active ingredient, soft gelatin capsules generally do not require a carrier. Suitable carriers for the manufacture of solutions and syrups are, for example, water, polyols, sucrose, invert sugar, glucose and the like. Suitable carriers for suppositories are, for example, natural or hardened oils, waxes, fats, semi-liquid or liquid polyols and the like.

The pharmaceutical preparations may also contain preservatives, solubilizers, stabilizers, wetting agents, emulsifiers, sweeteners, colorants, flavorants, salts for varying the osmotic pressure, buffers, masking agents or antioxidants. Other therapeutically valuable substances than the compounds of formula I may also be included.

Medicaments comprising a compound of formula I together with a compatible pharmaceutical carrier material are also an object of the present invention, as is a process for the manufacture of such medicaments, which comprises incorporating one or more of these compounds or salts thereof, and if desired one or more other therapeutically valuable substances, together with a compatible pharmaceutical carrier, into a galenical administration form.

As mentioned before, according to the invention, the compounds of formula I can be used as therapeutically active substances, in particular as anti-inflammatory agents or for the prevention of graft rejection after graft surgery. The dosage can vary within wide limits and will of course be adjusted in each particular case to the individual requirements. Generally, in the case of administration to an adult human, a daily dosage of from about 0.1mg/kg to about 100mg/kg, preferably from about 0.5mg/kg to about 5mg/kg, should be convenient. The daily dose may be administered in single or divided doses and, in addition, the upper limit of the dosage mentioned hereinbefore may be exceeded when treatment is found to be necessary.

Finally, the use of the compounds of the formula I for the preparation of medicaments, in particular for the treatment or prophylaxis of inflammatory, immunological, tumor, bronchopulmonary, dermatological and cardiovascular diseases, for the treatment of asthma, central nervous system diseases or diabetic complications or for the prevention of graft rejection after transplant surgery, is also an object of the present invention.

The compounds of formula I are useful in, but not limited to, the treatment of any disease or disorder in a human or other mammal that is exacerbated or caused by excessive or uncontrolled TNF or p38 kinase production in that mammal. Accordingly, the present invention provides a method of treating a cytokine mediated disease comprising administering a cytokine interfering effective amount of a compound of formula I, or a pharmaceutically acceptable salt or tautomer thereof.

The compounds of formula I are useful, but not limited to, treating inflammation in a subject, and as antipyretics for the treatment of fever. The compounds of the present invention are useful in the treatment of arthritis, including but not limited to rheumatoid arthritis, spondyloarthropathies, gouty arthritis, osteoarthritis, psoriatic arthritis, ankylosing spondylitis, systemic lupus erythematosus and juvenile arthritis, osteoarthritis, gouty arthritis and other arthritic conditions. Such compounds are useful in the treatment of pulmonary diseases or pneumonia, including adult respiratory distress syndrome, pulmonary sarcoidosis, asthma, silicosis, and chronic pneumonia. The compounds are also useful for treating viral and bacterial infections, including sepsis, septic shock, gram-negative sepsis, malaria, meningitis, infection or malignancy secondary to cachexia, Acquired Immune Deficiency Syndrome (AIDS) secondary to cachexia, AIDS, ARC (AIDS-related syndrome), pneumonia, and herpes virus. The compounds of the invention are also useful in the treatment of: bone resorption diseases such as osteoporosis, endotoxic shock, toxic shock syndrome, reperfusion injury, autoimmune diseases including graft-versus-host reaction and allograft rejection, cardiovascular diseases including myocardial infarction, atherosclerosis, thrombosis, congestive heart failure, cardiac reperfusion injury, renal reperfusion injury, liver disease and nephritis, and myalgia due to infection.

The compounds of the invention are also useful in the treatment of: alzheimer's disease, influenza, multiple sclerosis, cancer, diabetes, Systemic Lupus Erythematosus (SLE), skin-related diseases such as psoriasis, eczema, burns, dermatitis, scarring and scar tissue formation. In addition, the compounds of the present invention are useful in the treatment of gastrointestinal disorders such as inflammatory bowel disease, Crohn's disease, gastritis, irritable bowel syndrome and ulcerative colitis. The compounds of the present invention are also useful in the treatment of ocular diseases such as retinitis, retinopathies, uveitis, photophobia and acute injury to ocular tissues. The compounds of the invention may also be used in the treatment of: angiogenesis, including neoplasia; transferring; ophthalmic diseases such as corneal graft rejection, ocular neovascularization, retinal neovascularization including that following injury or infection, diabetic retinopathy, retrolental fibroplasia and neovascular glaucoma; ulcerative diseases such as gastric ulcer; pathological but non-malignant conditions such as hemangiomas, including neonatal hemangioma, nasopharyngeal angiofibroma, and avascular necrosis of bone; diabetic nephropathy and cardiomyopathy; and diseases of the female reproductive system, such as endometriosis. The compounds are also useful for preventing the production of cyclooxygenase-2 and have analgesic properties. Accordingly, the compounds of formula I are useful in the treatment of pain.

Other uses for compounds of formula I include the treatment of HCV, severe asthma, psoriasis, Chronic Obstructive Pulmonary Disease (COPD), and other diseases that may be treated with anti-TNF compounds.

In addition to being useful for the treatment of humans, the compounds of the present invention may also be useful for the veterinary treatment of animals such as companion animals, exotic and farm animals, including mammals, rodents, and the like. More preferred animals include horses, dogs and cats.

In addition, the compounds of the present invention may be used in the co-therapy, partially or completely replacing other conventional anti-inflammatory agents, for example with steroids, cyclooxygenase-2 inhibitors, NSAIDs, DMARDS, immunosuppressants, 5-lipoxygenase inhibitors, LTB₄Antagonists and LTA₄The hydrolase inhibitor is used together.

As used herein, the term "TNF-mediated disease" refers to any and all diseases and disorders in which TNF plays a role either through the control of TNF itself or through TNF causing the release of another monokine (such as, but not limited to, IL-1, IL-6 or IL-8). For example, in a disorder in which IL-1 is a major component and whose production or action is exacerbated or secreted in response to TNF, such disorders are thus considered TNF-mediated disorders.

The term "p 38 mediated disease" as used herein refers to any and all diseases and disorders that function either through the control of p38 itself or through p38 resulting in the release of another factor such as, but not limited to, IL-1, IL-6 or IL-8. For example, in a disorder in which IL-1 is a major component and its production or action is exacerbated or secreted in response to p38, such a disorder is therefore considered a p 38-mediated disorder.

Because TNF- β shares close structural homology with TNF- α (also known as cachectin), and because they both induce similar biological responses and bind to the same cellular receptors, the synthesis of both TNF- α and TNF- β is inhibited by the compounds of the present invention. Unless specifically stated otherwise, it is generally referred to as "TNF" in this specification.

Examples

Additional objects, advantages and novel features of the present invention will become apparent to those skilled in the art upon examination of the following illustrative examples, which are not intended to be limiting.

Example 1: preparation of 6- (2, 4-difluoro-phenoxy) -2- ((S) - (+) -2-hydroxy-1-methyl-ethylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (compound 16)

Step A: preparation of ethyl 4- ((S) -2-hydroxy-propylamino) -2-methylsulfanyl-pyrimidine-5-carboxylate

To a solution of 4-chloro-2-methylthiopyrimidine-5-carboxylic acid ethyl ester (Aldrich, 65g, 280mmol) at 0 deg.C in 500mL THF were added triethylamine (140mL 1000mmol) and (S) -1-amino-2-propanol (21g, 280 mmol). After stirring for 4 hours, water (200mL) was added and the phases were separated. The aqueous layer was extracted with dichloromethane. The organic phase was concentrated, the residue was dissolved in dichloromethane and washed with brine, dried over magnesium sulfate. Filtration and evaporation of the filtrate under reduced pressure gave 77g of ethyl 4- (S) -2-hydroxy-propylamino) -2-methylsulfanyl pyrimidine-5-carboxylate as a white solid.

And B: preparation of 4- ((S) -2-hydroxy-propylamino) -2-methylthiopyrimidine-5-methanol

Lithium aluminium hydride (5.7g, 150mmol) was stirred in dry THF (500mL) at 5 ℃ and treated dropwise with a solution of ethyl 4- ((S) -2-hydroxy-propylamino) -2-methylsulfanyl pyrimidine-5-carboxylate (27g, 100mmol) in dry THF (450 mL). The reaction mixture was stirred for 15 minutes, then water (18mL) was added dropwise carefully. The reaction was stirred for 30 minutes, then aqueous sodium hydroxide (15%, 8.5mL) was added dropwise, followed by water (25.5 mL). The resulting suspension was stirred at RT for 17 hours and then filtered. The filter residue was washed with isopropanol (2X, 100mL) and the combined filtrate and washings were evaporated under reduced pressure to give 25.8g of 4- ((S) -2-hydroxy-propylamino) -2-methylsulfanyl pyrimidine-5-methanol.

And C: preparation of 4- ((S) -2-hydroxy-propylamino) -2-methylsulfanyl-pyrimidine-5-carbaldehyde

4- ((S) -2-hydroxy-propylamino) -2-methylsulfanyl pyrimidine-5-methanol (26g, 100mmol) and 1L of dichloromethane were combined with stirring and treated with manganese dioxide (102g, 1 mol). The resulting suspension was stirred for 24 hours and then filtered through celite. The filter residue was washed with dichloromethane (100mL) and the combined filtrate and washings were evaporated under reduced pressure to give 16.5g of 4- ((S) -2-hydroxy-propylamino) -2-methylsulfanyl-pyrimidine-5-carbaldehyde as a white solid.

Sulfone

Step A: preparation of 6- (2, 4-difluorophenoxy) -8- ((S) -2-hydroxypropyl) -2-methylsulfanyl-8H-pyrido [2, 3-d ] pyrimidin-7-one

To a mixture of 4- ((S) -2-hydroxy-propylamino) -2-methylsulfanyl-pyrimidine-5-carbaldehyde (16.5g, 73mmol) and methyl (2, 4-difluorophenoxy) acetate (29.4g, 145mmol) in anhydrous dimethylformamide (300mL) was added potassium carbonate (30g, 218 mmol). The reaction mixture was heated to 60 ℃, cooled after 18 hours, and the dimethylformamide was distilled off under vacuum. The crude residue was suspended in water (300mL), extracted with dichloromethane, washed with brine and dried over magnesium sulfate. Filtration and concentration in vacuo afforded 41g of crude product which was subjected to silica gel column chromatography, eluting with 1% methanol in dichloromethane, to afford 30g of 6- (2, 4-difluorophenoxy) -8- ((S) -2-hydroxypropyl) -2-methylsulfanyl-8H-pyrido [2, 3-d ] pyrimidin-7-one (ms M +1 ═ 274).

And B: preparation of 6- (2, 4-difluorophenoxy) -8- ((S) -2-hydroxypropyl) -2-methanesulfonyl-8H-pyrido [2, 3-d ] pyrimidin-7-one

To a 5 ℃ solution of 6- (2, 4-difluorophenoxy) -8- ((S) -2-hydroxypropyl) -2-methylsulfanyl-8H-pyrido [2, 3-d ] pyrimidin-7-one (29.7g, 108mmol) in dichloromethane (500mL) was added m-chloroperbenzoic acid (55g, 240mmol) in portions and stirred for 24H. The reaction mixture was washed with aqueous sodium sulfite solution, aqueous sodium bicarbonate solution, and dried over magnesium sulfate. Filtration and evaporation gave 24g of 6- (2, 4-difluorophenoxy) -8- ((S) -2-hydroxypropyl) -2-methanesulfonyl-8H-pyrido [2, 3-d ] pyrimidin-7-one (ms M +1 ═ 412).

And C: preparation of 6- (2, 4-difluoro-phenoxy) -2- ((S) -2-hydroxy-1-methyl-ethylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one

To a solution of 6- (2, 4-difluorophenoxy) -8- ((S) -2-hydroxypropyl) -2-methanesulfonyl-8H-pyrido [2, 3-d ] pyrimidin-7-one (400mg, 1mmol) in THF (5mL) was added (S) -2-amino-1-propanol (0.38mL, 5mmol) and stirred at RT overnight. Concentration in vacuo, chromatography on silica gel eluting with 2% methanol in dichloromethane and conversion to the hydrochloride salt gave 320mg of 6- (2, 4-difluoro-phenoxy) -2- ((S) - (+) -2-hydroxy-1-methyl-ethylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (ms M +1 ═ 407, MP ═ 175.1-179.1 ℃).

Example 2: preparation of 6- (2, 4-difluoro-phenoxy) -2- (R) - (-) -2-hydroxy-1-methyl-ethylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (Compound 17)

To a solution of 6- (2, 4-difluorophenoxy) -8- ((S) -2-hydroxypropyl) -2-methanesulfonyl-8H-pyrido [2, 3-d ] pyrimidin-7-one (400mg, 1mmol) in THF (5mL) was added (R) -2-amino-1-propanol (0.38mL, 5mmol) and stirred at RT overnight. Concentrated in vacuo, chromatographed on silica gel, eluting with 2% methanol in dichloromethane, and converted to the hydrochloride salt to give 370mg of 6- (2, 4-difluoro-phenoxy) -2- ((R) -2-hydroxy-1-methyl-ethylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (ms spectrum M +1 ═ 407, MP ═ 174.9-178.1 ℃).

Example 3: preparation of 6- (2, 4-Difluorophenoxy) -2- (2-hydroxy-1, 1-dimethyl-ethylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (Compound 36)

To a solution of 6- (2, 4-difluorophenoxy) -8- ((S) -2-hydroxypropyl) -2-methanesulfonyl-8H-pyrido [2, 3-d ] pyrimidin-7-one (717mg, 1.74mmol) in THF (10mL) was added 2-amino-2-methyl-1-propanol (1.55g, 17.43mmol) and stirred at RT overnight. Concentration in vacuo and chromatography on silica gel eluting with 4% methanol in dichloromethane afforded 291mg of 6- (2, 4-difluorophenoxy) -2- (2-hydroxy-1, 1-dimethyl-ethylamino) -8- ((S) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one, after conversion to the hydrochloride salt, as a white solid (ms M +1 ═ 421, MP ═ 187.4-189.9 ℃).

Example 4: preparation of 6- (2, 4-difluoro-phenoxy) -2- ((S) -2-hydroxy-1-methyl-ethylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (compound 41)

Step A: preparation of ethyl 4- ((R) -2-hydroxy-propylamino) -2-methylsulfanyl-pyrimidine-5-carboxylate

To a solution of 4-chloro-2-methylthiopyrimidine-5-carboxylic acid ethyl ester (Aldrich, 62.6g, 269mmol) in 1L THF at 0 deg.C was added triethylamine (135mL 1000mmol) and (R) -1-amino-2-propanol (30g, 400 mmol). After stirring for 4 hours, evaporation under reduced pressure gave 66.6g of ethyl 4- (R) -2-hydroxy-propylamino) -2-methylsulfanyl pyrimidine-5-carboxylate as a white solid.

And B: preparation of 4- ((R) -2-hydroxy-propylamino) -2-methylthiopyrimidine-5-methanol

Lithium aluminum hydride (14g, 368mmol) was stirred in anhydrous THF (500mL) at 5 deg.C and treated dropwise with a solution of ethyl 4- (R) -2-hydroxy-propylamino) -2-methylthiopyrimidine-5-carboxylate (66.6g, 246mmol) in anhydrous THF (150 mL). The reaction mixture was stirred for 15 minutes, then water (18mL) was added dropwise carefully. The reaction was stirred for 30 minutes, then aqueous sodium hydroxide (15%, 8.5mL) was added dropwise followed by water (25.5 mL). The resulting suspension was stirred at RT for 17 hours and then filtered. The filter residue was washed with isopropanol (2X, 100mL) and the combined filtrate and washings were evaporated under reduced pressure to give 58.6g of 4- (R) -2-hydroxy-propylamino) -2-methylsulfanyl pyrimidine-5-methanol.

And C: preparation of 4- ((R) -2-hydroxy-propylamino) -2-methylsulfanyl-pyrimidine-5-carbaldehyde

4- (R) -2-hydroxy-propylamino) -2-methylsulfanyl pyrimidine-5-methanol (58.6g, 256mmol) and 1L of dichloromethane were combined with stirring and treated with manganese dioxide (222g, 2560 mol). The resulting suspension was stirred for 24 hours and then filtered through celite. The filter residue was washed with dichloromethane (100mL) and the combined filtrate and washings were evaporated under reduced pressure to give 34g of 4- ((R) -2-hydroxy-propylamino) -2-methylsulfanyl-pyrimidine-5-al as a white solid.

Sulfone

Step A: preparation of 6- (2, 4-difluorophenoxy) -8- ((R) -2-hydroxypropyl) -2-methylsulfanyl-8H-pyrido [2, 3-d ] pyrimidin-7-one

To a mixture of 4- ((R) -2-hydroxy-propylamino) -2-methylsulfanyl-pyrimidine-5-carbaldehyde (17.7g, 78mmol) and methyl (2, 4-difluorophenoxy) acetate (31.6g, 156mmol) in anhydrous dimethylformamide (300mL) was added potassium carbonate (30g, 218 mmol). The reaction mixture was heated to 60 ℃, cooled after 18 hours, and the DMF was distilled off. The residue was suspended in water (300mL), extracted with dichloromethane, washed with brine and dried over magnesium sulfate. Filtration and concentration in vacuo afforded 29.5g of crude product which was subjected to silica gel column chromatography, eluting with 1% methanol in dichloromethane, to afford 17.5g of 6- (2, 4-difluorophenoxy) -8- ((R) -2-hydroxypropyl) -2-methylsulfanyl-8H-pyrido [2, 3-d ] pyrimidin-7-one (ms M +1 ═ 274).

And B: preparation of 6- (2, 4-difluorophenoxy) -8- ((R) -2-hydroxypropyl) -2-methanesulfonyl-8H-pyrido [2, 3-d ] pyrimidin-7-one

To a 5 ℃ solution of 6- (2, 4-difluorophenoxy) -8- ((R) -2-hydroxypropyl) -2-methylsulfanyl-8H-pyrido [2, 3-d ] pyrimidin-7-one (9.38g, 24.7mmol) in dichloromethane (200mL) was added m-chloroperbenzoic acid (12.5g, 54mmol) in portions and stirred for 24H. The reaction mixture was washed with aqueous sodium sulfite solution and aqueous sodium bicarbonate solution, and dried over magnesium sulfate. Filtration and evaporation gave 10.7g of 6- (2, 4-difluorophenoxy) -8- ((R) -2-hydroxypropyl) -2-methanesulfonyl-8H-pyrido [2, 3-d ] pyrimidin-7-one (ms M +1 ═ 412).

And C: preparation of 6- (2, 4-difluoro-phenoxy) -2- ((S) -2-hydroxy-1-methyl-ethylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one

To a solution of 6- (2, 4-difluorophenoxy) -8- ((R) -2-hydroxypropyl) -2-methanesulfonyl-8H-pyrido [2, 3-d ] pyrimidin-7-one (615mg, 1.5mmol) in THF (5mL) was added (S) -2-amino-1-propanol (1.2mL, 15mmol) and stirred at RT overnight. Concentrated in vacuo, chromatographed on silica gel, eluted with 2% methanol in dichloromethane and converted to the hydrochloride salt to give 295mg of 6- (2, 4-difluoro-phenoxy) -2- ((S) -2-hydroxy-1-methyl-ethylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (ms spectrum M +1 ═ 407, MP ═ 186.0-189.1 ℃.

Example 5: preparation of 6- (2, 4-difluoro-phenoxy) -2- (R) -2-hydroxy-1-methyl-ethylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (Compound 48)

To a solution of 6- (2, 4-difluorophenoxy) -8- ((R) -2-hydroxypropyl) -2-methanesulfonyl-8H-pyrido [2, 3-d ] pyrimidin-7-one (400mg, 1mmol) in THF (5mL) was added (R) -2-amino-1-propanol (0.38mL, 5mmol) and stirred at RT overnight. Concentrated in vacuo, chromatographed on silica gel, eluted with 2% methanol in dichloromethane and converted to the hydrochloride salt to give 350mg of 6- (2, 4-difluoro-phenoxy) -2- ((R) -2-hydroxy-1-methyl-ethylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (ms M +1 ═ 407, MP ═ 181.5-184.4 ℃).

Example 6: preparation of 6- (2, 4-Difluorophenoxy) -2- (2-hydroxy-1, 1-dimethyl-ethylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (Compound 31)

A mixture of 6- (2, 4-difluorophenoxy) -8- ((R) -2-hydroxypropyl) -2-methanesulfonyl-8H-pyrido [2, 3-d ] pyrimidin-7-one (886mg, 2.15mmol) and 2-amino-2-methyl-1-propanol (5.15g, 58mmol) was heated under nitrogen at 60 ℃ for 2 hours. Cooling and chromatography on silica gel eluting with 2% methanol in dichloromethane afforded 385mg 6- (2, 4-difluorophenoxy) -2- (2-hydroxy-1, 1-dimethyl-ethylamino) -8- ((R) -2-hydroxy-propyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one after conversion to the hydrochloride salt (ms M +1 ═ 421, MP ═ 182.0-183.9 ℃).

Example 7: preparation of 6- (2, 4-Difluorophenoxy) -2- ((S) -2-hydroxy-1-methyl-ethylamino) -8- (2-oxopropyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (Compound 45)

Step a: preparation of 6- (2, 4-difluorophenoxy) -2-methanesulfonyl-8- (2-oxopropyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one

To a solution of oxalyl chloride (1.05mL, 12mmol) at-60 deg.C in dichloromethane (100mL) was added dimethyl sulfoxide (1.7mL, 24mmol) and 6- (2, 4-difluorophenoxy) -8- (2-hydroxypropyl) -2-methanesulfonyl-8H-pyrido [2, 3-d ] pyrimidin-7-one (4.12g, 10 mmol). To the mixture was added triethylamine (7mL, 50mmol) and stirred overnight. Water (100mL) was added, extracted with dichloromethane, washed with brine, and dried over magnesium sulfate. Filtration and concentration in vacuo, chromatography on silica gel eluting with 2% methanol in dichloromethane afforded 1.0g of 6- (2, 4-fluorophenoxy) -2-methanesulfonyl-8- (2-oxopropyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (ms M +1 ═ 410).

Step b: preparation of 6- (2, 4-Difluorophenoxy) -2- ((S) -2-hydroxy-1-methyl-ethylamino) -8- (2-oxopropyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (Compound 45)

To a suspension of 6- (2, 4-difluorophenoxy) -2-methanesulfonyl-8- (2-oxopropyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (412mg, 1mmol) in THF at RT was added (S) -2-amino-1-propanol (0.39mL, 5mmol) and stirred overnight. Concentrated in vacuo and chromatographed on silica gel, eluting with 2% methanol in dichloromethane, to give, after conversion to the hydrochloride, 330mg of 6- (2, 4-difluorophenoxy) -2- ((S) -2-hydroxy-1-methyl-ethylamino) -8- (2-oxopropyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (ms spectrum M +1 ═ 405, MP ═ 207.9-214.6 ℃).

Example 8: preparation of 6- (2, 4-Difluorophenoxy) -2- ((R) -2-hydroxy-1-methyl-ethylamino) -8- (2-oxopropyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (Compound 49)

To a suspension of 6- (2, 4-difluorophenoxy) -2-methanesulfonyl-8- (2-oxopropyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (417mg, 1mmol) in THF (10mL) at RT was added (R) - (-) -2-amino-1-propanol (0.40mL, 5mmol) and stirred overnight. Concentrated in vacuo and chromatographed on silica gel, eluting with 2% methanol in dichloromethane, to give, after conversion to the hydrochloride, 330mg of 6- (2, 4-difluorophenoxy) -2- ((R) -2-hydroxy-1-methyl-ethylamino) -8- (2-oxopropyl) -8H-pyrido [2, 3-d ] pyrimidin-7-one (ms M +1 ═ 405, MP ═ 207.8-216.4 ℃).

Example 9: in vitro assay

The in vitro p38MAP kinase inhibitory activity of the compounds of the invention was determined using Ahn et al, j.biol.chem.vol.266: 4220-4227(1991) by measuring gamma-phosphate from gamma-via p-38 kinase³³Transfer of P-ATP to Myelin Basic Protein (MBP).

The phosphorylated form of recombinant p38MAP kinase was co-expressed in E.coli using SEK-1 and MEKK (see, Khokhlatchev et al, J.biol.chem.Vol.272: 11057-11062(1997)) and then purified by affinity chromatography on a nickel column.

Phosphorylated p38MAP kinase was dissolved in kinase buffer (20mM 3- (N-morpholino) propanesulfonic acid, pH7.2, 25mM beta-phosphoglyceride, 5mM ethylene glycol-bis (. beta. -aminoethylether) -N, N, N ', N' -tetraacetic acid, 1mM sodium orthovanadate, 1mM dithiothreitol, 40mM magnesium chloride) And (4) diluting. Test compounds dissolved in DMSO or DMSO alone (control) were added and the samples were incubated at 30 ℃ for 10 minutes. By adding MBP and gamma-³³The substrate mixture of P-ATP initiates the kinase reaction. After an additional 20 minutes incubation at 30 ℃, the reaction was stopped by adding 0.75% phosphoric acid. From the residual gamma-ion by using phosphocellulose membrane (Millipore, Bedford, MA)³³Phosphorylated MBP was isolated from P-ATP and quantified using a scintillation counter (Packard, Meriden, CT).

Using the above assay, the compounds of the invention are shown to be p38MAP kinase inhibitors. P38IC exhibited by the compounds of the invention₅₀The value is less than 0.001 to 0.1. mu.M. For example, 6- (2, 4-difluoro-phenoxy) -8- (3-hydroxy-propyl) -2- (tetrahydropyran-4-ylamino) -8H-pyrido [2, 3-d]IC of pyrimidin-7-one in the above assay₅₀The concentration was 0.0008. mu.M.

Example 10: in vitro assay

This example illustrates an in vitro assay of Human Whole Blood (HWB) (i.e., by inhibiting LPS-induced IL-1 β production of p38MAP kinase in undiluted human whole blood) for evaluating the results of comparison of compounds of the invention and the corresponding alkyl analogs.

LPS (lipopolysaccharide) treatment of human whole blood induces IL-1 β (interleukin-1 β) production, and IL-1 β production can be measured by IL-1 β specific ELISA. Human whole blood was preincubated with a solution of the indicated concentration of the compound of the invention in 0.5% DMSO (final concentration) for 30 minutes at 37 ℃. Samples were stimulated with 0.5 μ g/mL lipopolysaccharide (LPS, obtained from Sigma) (final concentration) for 18 hours to induce the synthesis and secretion of IL-1 β as measured by IL-1 β ELISA.

Compound solution

Compounds were dissolved in 449 μ L DMSO to prepare 6mM stock solutions in DMSO (obtained from Sigma). Starting from a 6mM stock, 6 semilog serial dilutions were made in DMSO to give the following concentrations: 1.9mM, 600, 190, 60, 19 and 6. mu.M. Tubes 1-7 are labeled. 6mM stock solution was placed in tube 1. 216 μ L of DMSO was placed in each of tubes 2-7. Transfer 100. mu.L from tube 1 to tube 2. Tube 2 was rotated and 100. mu.L was transferred from tube 2 to tube 3. This process is repeated until the tube 7.

Additional dilutions 1/20(10 μ L dilution to 190 μ L RPMI 1640 medium, obtained from Gibco-BRL) were made using serial DMSO dilutions of the compound prepared above, resulting in the final compound concentration curve: 30, 10, 2.9, 1, 0.3, 0.1, 0.03 μ M.

LPS solution

And (3) LPS recombination: in a vial of 10mg LPS, 10mL of 1X phosphate-buffered saline (i.e., 1XPBS, from Gibco-BRL) was added, mixed well and transferred to a 50mL tube. An additional 10mL was added to the LPS vial, followed by rinsing, which was added to the 50mL tube and mixed. The solution was filtered and sterilized and aliquoted into the required amounts (100 μ Ι _ aliquots are sufficient for 4 plates). This gave a 0.5mg/mL stock solution which was diluted 1/100 for use in the protocol. Immediately prior to use, stock solution of LPS 1/100 was diluted (100 μ Ι _ in 10mL RPMI).

Test procedure

The assay was performed on a 96-well U-plate (obtained from Costar). Two controls, which contained no compound, with or without LPS, were included in each experiment. All samples and controls were replicated three times.

Human blood (from donors who did not receive the drug for at least 14 days and did not drink for 48 hours) was collected into siliconized vacutainers containing heparin (19 units/ml). A 25 μ L aliquot of 5% DMSO in RPMI 1640 was added to control wells (with or without LPS control). A 25 μ L aliquot of each compound concentration prepared as above was dispensed into designated wells. mu.L of human whole blood was added to each well and incubated at 37 ℃ and 5% CO₂Incubate under conditions for 30 minutes. 25 μ L of diluted LPS was dispensed to all wells except the control wells containing no LPS. To control wells without LPS 25 μ L RPMI was added.

The plates were incubated at 37 ℃ and 5% CO₂Incubate under conditions for 18 hours. After incubation, plates were centrifuged at 400Xg to allowCells were pelleted and plasma was collected, at which time care should be taken not to disturb the pellet. The plasma was transferred to a new 96-well polypropylene plate. The ELISA was immediately performed and the remaining plasma was stored at-20 ℃ for re-testing when required.

ELISA draft

The IL-1 β ELISA used two anti-IL-1 β monoclonal antibodies: IL β 1-H6(1mg/mL) and IL β 1-H67(2.71 mg/mL).

Material

Recombinant human IL-1 beta (rhuIL-1 beta, 2.5. mu.g/mL) was obtained from R&D Systems. Phosphate buffered saline-Dulbecco's (1XPBS) was obtained from Gibco-BRL. Phosphate buffered saline (10XPBS) was obtained from Gibco-BRL. Dulbecco's variant without calcium and magnesium, pH 7.2. Unopened bottles were stored at RT. ELISA Incubation Buffer (EIB): -0.1% BSA/PBS; 1g Bovine Serum Albumin (BSA); 100mL 10 XPBS; deionized water was added to 1 liter and stored at 4 ℃. ELISA Wash Buffer (EWB): 0.05% Tween/PBS; 0.5mL Tween 20; 100mL10xPBS; deionized water was added to 1 liter and stored at 4 ℃. Blocking buffer-3% skim milk powder/PBS: 15g of skimmed milk powder (Carnation); 50mL 10 XPBS; deionized water was added to 500mL and stored at 4 ℃. Peroxidase conjugated streptavidin (obtained from Pharmingen): dilute approximately 1: 3000 (10. mu.L/30 mL) in EWB buffer. 0.1M citrate buffer, ph 4.5: 9.6g citric acid (MW 192.1, obtained from Sigma); 14.7g trisodium citrate (MW 294.1, obtained from Sigma); adjusted to pH4.5 with NaOH and distilled water was added to 500 mL. Storage at 4 ℃. OPD substrate solution: 1mg/mL OPD/0.03% H₂O₂A citrate buffer; 1 piece of o-phenylenediamine (OPD, from Zymed); 12 μ L of 30% hydrogen peroxide; 12mL of 0.1M citrate buffer.

Preparation of standards (freshly prepared before placement on the plate).

A standard curve was constructed using rhuIL-1. beta. stock solution (2.5. mu.g/mL). The concentration of the curve is: 12500, 4167, 1389, 463, 154, 51 and 17 pg/mL. The tubes are numbered 1-8. rhuIL-1. beta. stock was diluted 1/500 in tube 1 (3. mu.L stock + 597. mu.L EWB). 400 μ L of EWB was dispensed into tubes 2-8. 200 μ L was transferred from tube 1 to tube 2 and spun. Transfer 200. mu.L from tube 2 to tube 3. This operation is repeated until the tube 7. Tube 8 was used as ELISA assay blank.

Plasma samples were diluted 1: 4 in EWB (20. mu.L plasma + 60. mu.L EWB).

Preparation of antibody solutions

Antibody IL β 1-H6 was diluted 1/100 in 1XPBS to give a 10 μ g/mL solution. For each plate, 50 μ L of antibody was diluted in 5mL PBS. Antibody IL β 1-H671/100 was diluted in EWB to give a 2 μ g/mL solution. For each plate, 3.69 μ L of antibody was diluted in 5 mLEIB.

Procedure

A96-well EIA plate was coated with 50. mu.L of antibody IL β 1-H6 (10. mu.g/mL) per well, gently shaken to remove any air bubbles, sealed with a plate sealer and incubated overnight at 4 ℃ in a humidified chamber. The board was emptied and patted dry on a non-fluffed paper towel. Non-specific binding sites were blocked with 175. mu.L of blocking buffer per well for 1-2 hours at RT. The plate was washed once with EWB (i.e., empty plate, filled with 150 μ L EWB, emptied and patted dry on a lint free paper towel).

Triplicate aliquots of 25 μ L of the standard were added to the appropriate wells. (each plate has its own standard curve). A 25 μ L aliquot of diluted plasma was added to the appropriate wells. To all wells 25. mu.L of biotinylated monoclonal antibody IL β 1-H67(2. mu.g/mL) was added. The plate was sealed with a plate sealer and incubated at RT for 2 hours (or overnight at 4 ℃) with gentle shaking (Bellco Mini-Orbital Shaker, setting 3.5). After incubation, plates were washed 3x with EWB (as described above). A50. mu.L aliquot of peroxidase-streptavidin diluted 1: 3000 in EIB was added to each well. The plate was sealed with a plate sealer and incubated at RT for 1 hour while shaking and washed 3x as described above.

Mixing OPD tabletsDissolved in citrate buffer (1 tablet/12 mL citrate buffer), and 12. mu.L of 30% H was added to OPD/citrate buffer₂O₂. 50 μ L of OPD substrate solution was dispensed to each well and the plates were incubated in the dark at RT for 30 minutes for visualization. Read plate at dual wavelength: sample filter 450 nm/reference filter 650 nm. A standard curve (absorbance vs. concentration) was made using the values of the samples containing the standards for determining the concentration of the unknown sample.

Statistical method

If neither end of the concentration-inhibition curve contains a 50% point, the IC will be₅₀Reported as > highest concentration or < lowest concentration. In addition, if the concentration number is ≧ 5, the data is fit to the following 2-parameter model to estimate IC₅₀：

The model assumes minimum and maximum responses of 0% and 100%, respectively, and estimates the IC₅₀And a slope parameter. If the non-linear regression fails or if the test concentration is < 5, IC is estimated by linear regression using 2 points located 50% of the side₅₀。

If linear regression is used to estimate IC₅₀This is output in the form of a test record, which can be seen as an IC₅₀The presence (non-linear regression) or absence (linear regression) of the standard error and slope parameters.

Using the above assay, the compounds of the invention show an inhibitory effect on LPS-induced IL-1 β production in undiluted human whole blood by inhibiting p38MAP kinase, which, as described above, mediates IL-1 β production. Compounds of the invention exhibit IC for LPS-induced IL-1 β production in undiluted human whole blood₅₀Values ranged from < 0.001. mu.M to 0.30. mu.M. For example, 6- (2, 4-difluoro-phenoxy) -8- ((R) -2-hydroxy-propyl) -2- (tetrahydropyran-4-ylamino) -8H-pyrido [2, 3-d]Pyrimidin-7-one shows an IC50 of 0.001. mu.M.

Surprisingly, use is made of compounds in which R is present in the formula (I)²The compounds of the present invention which are hydroxyalkyl or alkoxyalkyl have a substantially greater inhibitory effect on LPS-induced IL-1 β production than compounds wherein R is²Corresponding compounds which are methyl or other alkyl groups. This unexpected advantage of the present invention is more fully illustrated in table 2, where R is²Representative compounds of the invention which are hydroxyalkyl (of formula I) and wherein R²The corresponding analog is methyl for comparison). The compounds in the first or leftmost column of table 1 were prepared as described in the examples of this specification and are also shown in table 1. The compounds in the second or central column of table 2 were prepared according to the procedure reported in WO 02/064594. The numerical values in the third or rightmost column correspond to their ratios:

(IC for IL-1. beta. production₅₀Inhibition rate of R²IC formed by hydroxyalkyl)/(IL-1. beta₅₀Inhibition rate of R²Methyl).

As can be seen from Table 2, where R²Compounds which are hydroxyalkyl groups inhibit LPS-induced IL-1 β production in undiluted human whole blood in comparison to the corresponding methyl analogue (R)²Methyl) is 2.7-fold to > 100-fold greater, i.e., 270% to > 10,000%.

TABLE 2

The foregoing discussion of the invention has been presented for purposes of illustration and description. The foregoing is not intended to limit the invention to the form disclosed herein. While the description of the invention has included one or more embodiments and certain variations and modifications, for example, other variations and modifications are within the scope of the invention, as they are within the skill and knowledge of those in the art, after understanding the present disclosure. It is intended to obtain rights which include alternative embodiments to the extent permitted, including alternate, interchangeable and/or equivalent structures, functions, ranges or steps to those claimed, whether or not such alternate, interchangeable and/or equivalent structures, functions, ranges or steps are disclosed herein, and without intending to expressly provide any patentable subject matter. For all of these purposes, the entire disclosures of all publications, patents and patent applications cited herein are hereby incorporated by reference.

Claims (6)

1. A compound of formula I

Wherein

X¹Is O, C ═ O or S (O)_nWherein n is 0, 1 or 2;

Ar¹is aryl or heteroaryl;

R¹is alkoxyalkyl, alkyl, cycloalkyl, cyclicAn alkyl, heterocyclyl, hydroxyalkyl or hydroxycycloalkyl group; and is

R²Is hydroxyalkyl, oxoalkyl or hydroxycycloalkyl;

wherein the alkyl refers to a linear saturated monovalent hydrocarbon group of 1 to 6 carbon atoms or a branched saturated monovalent hydrocarbon group of 3 to 6 carbon atoms;

cycloalkyl refers to a saturated monovalent cyclic hydrocarbon group of 3 to 7 ring carbons, optionally substituted with 1, 2, or 3 substituents selected from alkyl, hydroxy, alkoxy, haloalkyl, haloalkoxy, halo, amino, monoalkylamino, dialkylamino, and acyl;

aryl means a monovalent monocyclic or bicyclic aromatic hydrocarbon group optionally independently substituted with 1, 2 or 3 substituents selected from the group consisting of alkyl, hydroxy, alkoxy, haloalkyl, haloalkoxy, halo, nitro, cyano, amino, monoalkylamino, dialkylamino, methylenedioxy, ethylenedioxy and acyl;

heteroaryl refers to a monovalent monocyclic or bicyclic group of 5-12 ring atoms containing at least one aromatic ring, wherein the aromatic ring contains 1, 2 or 3 ring heteroatoms selected from N, O or S, the remaining ring atoms are C, the connecting point of the heteroaryl is on the aromatic ring, the heteroaryl is optionally and independently substituted by 1 or 2 substituents selected from alkyl, haloalkyl, hydroxyl, alkoxy, halo, nitro or cyano;

heterocyclyl is a saturated or unsaturated non-aromatic cyclic group of 3 to 8 ring atoms, 1 or 2 of which are selected from N, O or S (O)_nWherein n is an integer of 0 to 2, the remaining ring atoms are C, wherein 1 or 2 carbon atoms are optionally substituted with carbonyl, the heterocyclyl ring is optionally independently substituted with 1, 2 or 3 substituents selected from alkyl, haloalkyl, hydroxyalkyl, halo, hydroxy, alkoxy, amino, monoalkylamino, dialkylamino, aralkyl, and-S (O)_nR^kWherein n is an integer of 0 to 2, such that when n is 0, R^kIs hydrogen, alkyl, cycloalkyl or cycloalkylalkyl, and when n is 1 or 2, R is^kIs alkyl, cycloalkyl, cycloalkylalkyl, amino, acylAmino, monoalkylamino or dialkylamino.

2. A compound of formula I according to claim 1, having formula II

Wherein

m is 0 to 4;

each R³Is alkyl, halogen, alkoxy or haloalkyl; and is

R¹And R²As defined in claim 1.

3. A process for the preparation of a compound of formula I according to claim 1, which process comprises reacting a compound of formula If

Wherein R is C₁-C₆Alkyl radical, and X¹、Ar¹And R²Has the meaning as defined in claim 1, and the formula R¹-NH₂In which R is¹Have the meaning as defined in claim 1.

4. The use of a compound of formula I according to claim 1 for the manufacture of a medicament for the control or prevention of diseases mediated by p38 kinase.

5. The use according to claim 4, wherein said p38 kinase mediated disease is arthritis, Crohn's disease, irritable bowel syndrome, adult respiratory distress syndrome, chronic obstructive pulmonary disease or Alzheimer's disease.

6. A composition comprising a pharmaceutically acceptable excipient and a compound of formula I according to claim 1.