AU741033B2 - 5,6,7-trisubstituted-4-aminopyridol(2,3-d)pyrimidine compounds - Google Patents

Description

5,6,7-Trisubstituted-4-aminopyrido[2,3-d]pyrimidine Compounds Technical Field The present invention relates a method for inhibiting adenosine kinase by administering 5,6,7trisubstituted-4-aminopyrido[2,3-d]pyrimidine compounds, to pharmaceutical compositions containing such compounds, as well as novel 5,6,7-trisubstituted-4-aminopyrido[2,3-d]pyrimidine compounds.

Background of the Invention Adenosine kinase (ATP:adenosine 5'-phosphotransferase, EC 2.7.1.20) is a ubiquitous enzyme which catalyses the phosphorylation of adenosine to AMP, using ATP, preferentially, as the phosphate source. Adenosine kinase has broad tissue and species distribution, and has been isolated from yeast, a variety of mammalian sources and certain microorganisms. It has been found to be present in virtually every human tissue assayed including kidney, liver, brain, spleen, placenta and pancreas. Adenosine kinase is a key enzyme in the control of the cellular concentrations of adenosine.

Adenosine is a purine nucleoside that is an intermediate in the pathways of purine nucleotide degradation and salvage. Adenosine also has many important physiological effects, many of which are mediated through the activation of specific ectocellular receptors, termed Pi receptors (Burnstock, in Cell Membrane Receptors for Drugs and Hormones, 1978, (Bolis and Straub, eds.) Raven, New York, pp. 107-118; Fredholm, et al., Pharmacol. Rev. 1994, 46: 143-156.

S In the central nervous system, adenosine inhibits the release of certain neurotransmitters 20 (Corradetti, et al., Eur. J. Pharmacol. 1984, 104: 19-26), stabilises membrane potential (Rudolphi, et al., Cerebrovasc. Brain Metab. Rev. 1992, 4: 346-360), functions as an endogenous anticonvulsant (Dragunow, Trends Pharmacol. Sci. 1986, 7: 128-130) and may have a role as an endogenous neuroprotective agent (Rudolphi, et al., Trends Pharmacol. Sci., 1992, 13: 439-445). Adenosine may play a role in several disorders of the central nervous system such as schizophrenia, anxiety, 25 depression and Parkinson's disease. (Williams, in Psychopharmacology: The Fourth Generation of Progress; Bloom, Kupfer Raven Press, New York, 1995, pp 643-655.

Adenosine has also been implicated in modulating transmission in pain pathways in the spinal cord (Sawynok, et al., Br. J. Pharmacol., 1986, 88: 923-930), and in mediating the analgesic effects of morphine (Sweeney, et J. Pharmacol. Exp. Ther. 1987, 243: 657-665).

In the immune system, adenosine inhibits certain neutrophil functions and exhibits antiinflammatory effects (Cronstein, J. Appl. Physiol. 1994, 76: 5-13). An AK inhibitor has been reported to decrease paw swelling in a model of adjuvant arthritis in rats (Firestein, et al., Arthritis and Rheumatism, 1993, 36, S48.

Adenosine also exerts a variety of effects on the cardiovascular system, including vasodilation, impairment of atrioventricular conduction and endogenous cardioprotection in myocardial ischaemia and reperfusion (Mullane and Williams, in Adenosine and Adenosine Receptors, 1990 (Williams, ed.) Humana Press, New Jersey, pp. 289-334). The widespread actions of adenosine also include effects on the renal, respiratory, gastrointestinal and reproductive systems, as well as on blood cells and adipocytes. Adenosine, via its Al receptor activation on adipocytes, plays a role in diabetes by )STORinhibiting lipolysis [Londos, et al., Proc. Natl. Acad. Sci. USA, 1980, 77, 2551.

LibC/475183speci ^07 Endogenous adenosine release appears to have a role as a natural defence mechanism in various pathophysiologic conditions, including cerebral and myocardial ischaemia, seizures, pain, inflammation and sepsis. While adenosine is normally present at low levels in the extracellular space, its release is locally enhanced at the site(s) of excessive cellular activity, trauma or metabolic stress.

Once in the extracellular space, adenosine activates specific extracellular receptors to elicit a variety of responses which tend to restore cellular function towards normal (Bruns, Nucleosides Nucleotides, 1991, 10: 931-943; Miller and Hsu. J. Neurotrauma, 1992, 9: S563-S577). Adenosine has a half-life measured in seconds in extracellular fluids (Moser, et al., Am. J. Physiol. 1989, 25: C799-C806), and its endogenous actions are therefore highly localised.

The inhibition of adenosine kinase can result in augmentation of the local adenosine concentrations at foci of tissue injury, further enhancing cytoprotection. This effect is likely to be most pronounced at tissue sites where trauma results in increased adenosine production, thereby minimising systemic toxicities.

Pharmacological compounds directed towards adenosine kinase inhibition provide potentially effective new therapies for disorders benefited by the site- and event-specific potentiation of adenosine. Disorders where such compounds may be useful include ischaemic conditions such as cerebral ischaemia, myocardial ischaemia, angina, coronary artery bypass graft surgery (CABG), percutaneous transluminal angioplasty (PTCA), stroke, other thrombotic and embolic conditions, and Sneurological disorders such as epilepsy, anxiety, schizophrenia, nociperception including pain 20 perception, neuropathic pain, visceral pain, as well as inflammation, arthritis, immunosuppression, sepsis, diabetes and gastrointestinal dysfunctions such as abnormal gastrointestinal motility.

A number of compounds have been reported to inhibit adenosine kinase. The most potent of these include 5'-amino-5'-deoxyadenosine (Miller, et al., J. Biol. Chem. 1979, 254: 2339-2345), iodotubercidin (Wotring and Townsend, Cancer Res. 1979, 39: 3018-3023) and 25 iodotubercidin (Davies, et al., Biochem. Pharmacol. 1984, 33: 347-355).

Adenosine kinase is also responsible for the activation of many pharmacologically active nucleosides (Miller, et al., J. Biol. Chem. 1979. 254: 2339-2345), including tubercidin, formycin, ribavirin, pyrazofurin and 6-(methylmercapto)purine riboside. These purine nucleoside analogues represent an important group of antimetabolites which possess cytotoxic, anticancer and antiviral properties. They serve as substrates for adenosine kinase and are phosphorylated by the enzyme to generate the active form. The loss of adenosine kinase activity has been implicated as a mechanism of cellular resistance to the pharmacological effects of these nucleoside analogues (eg. Bennett, et al., Mol. Pharmacol., 1966, 2: 432-443; Caldwell, et al., Can. J. Biochem., 1967, 45: 735-744; Suttle, et al., Europ. J. Cancer, 1981, 17: 43-51). Decreased cellular levels of adenosine kinase have also been associated with resistance to the toxic effects of 2'-deoxyadenosine (Hershfield and Kredich, Proc.

Natl. Acad. Sci. USA, 1980, 77: 4292-4296). The accumulation of deoxyadenosine triphosphate (dATP), derived from the phosphorylation of 2'-deoxyadenosine, has been suggested as a toxic mechanism in the immune defect associated with inheritable adenosine deaminase deficiency (Kredich and Hershfield, in The Metabolic Basis of Inherited Diseases, 1989 (Scriver, et al., eds.), 6. .KGpraw-Hill, New York, pp. 1045-1075).

LibC/475183speci B.S. Hurlbert et al. Med. Chem., 11: 711-717 (1968)) disclose various 2,4-diaminopyrido[2,3d]pyrimidine compounds having use as antibacterial agents. R. K. Robins et al. Amer. Chem.

Soc., 80:3449-3457 (1958)) disclose methods for preparing a number of 2,4-dihydroxy-, 2,4-diamino-, 2-amino-4-hydroxy- and 2-mercapto-4-hydroxypyrido[2,3-d]pyrimidines having antifolic acid activity.

R. Sharma et al., (Indian J. Chem., 311B: 719-720 (1992)) disclose 4-amino-5-(4-chlorophenyl)-7-(4nitrophenyl)pyrido[2,3-d]pyrimidine and 4-amino-5-(4-methoxyphenyl)-7-(4-nitrophenyl)pyrido[2,3d]pyrimidine compounds having antibacterial activity. A. Gupta et al., Indian Chem. Soc., 71: 635- 636 (1994)) disclose 4-amino-5-(4-fluorophenyl)-7-(4-fluorophenyl)pyrido[2,3-d]pyrimidine and 4amino-5-(4-chlorophenyl)-7-(4-fluorophenyl)pyrido[2,3-d]pyrimidine compounds having antibacterial activity. L. Prakash et al., Pharmazie, 48: 221-222 (1993)) disclose 4-amino-5-phenyl-7-(4aminophenyl)pyrido[2,3-d]pyrimidine, 4-amino-5-phenyl-7-(4-bromophenyl)pyrido[2,3-d]pyrimidine, 4amino-5-(4-methoxyphenyl)-7-(4-aminophenyl)pyrido[2,3-d]pyrimidine, and 4-amino-5-(4methoxyphenyl)-7-(4-bromophenyl)pyrido[2,3-d]pyrimidine compounds having antifungal activity. P.

Victory et al., Tetrahedron, 51:10253-10258 (1995)) disclose the synthesis of 4-amino-5,7diphenylpyrido[2,3-d]pyrimidine compounds from acyclic precursors. Bridges et al. PCT application WO 95/19774, published July 27, 1995) disclose various bicyclic heteroaromatic compounds as having utility for inhibiting tyrosine kinase of epidermal growth factors.

SSummary of the Invention :I The present invention provides for 5,6,7-trisubstituted-4-aminopyrido[2,3-d]pyrimidine 20 compounds having utility as adenosine kinase inhibitors.

In one aspect, the present invention provides compounds having the formula (I)

NH

2

R

1 ~N R 2 N R3 (I) wherein R 1 is phenyl or thienyl, each of which is unsubstituted or substituted with one or two substituents independently selected from alkyl, alkoxy or halo; R 2 is lower alkyl, cycloalkyl, phenyl or 25 phenyl alkyl wherein each of said phenyl is unsubstituted or substituted with one or two substituents independently selected from alkyl, alkoxy or halo; and R 3 is phenyl, pyridyl or thienyl, each of which is unsubstituted or substituted with one or two substituents selected from alkoxy or di(loweralkyl)amino.

In another aspect, the present invention provides a method for inhibiting adenosine kinase by administering a compound of formula In particular, the method of inhibiting adenosine kinase comprises exposing an adenosine kinase to an effective inhibiting amount of a compound of Formula I of the present invention. Where the adenosine kinase is located in vivo, the compound is administered to the organism.

In yet another aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula I above in combination with a pharmaceutically acceptable carrier.

In still another aspect, the present invention provides a method of treating ischaemia, S neurological disorders, nociperception, inflammation, immunosuppression, gastrointestinal LibC/475183speci dysfunctions, diabetes and sepsis in a mammal in need of such treatment, comprising administering to the mammal a therapeutically effective amount of a compound of Formula I of the present invention.

In a preferred aspect, the present invention provides a method of treating cerebral ischaemia, myocardial ischaemia, angina, coronary artery bypass graft surgery, percutaneous transluminal angioplasty, stroke, thrombotic and embolic conditions, epilepsy, anxiety, schizophrenia, pain perception, neuropathic pain, visceral pain, arthritis, sepsis, diabetes and abnormal gastrointestinal motility in a mammal in need of such treatment, comprising administering to the mammal a therapeutically effective amount of a compound of Formula I of the present invention.

The present invention also contemplates the use of pharmaceutically acceptable salts and amides of the compounds of Formula I.

In another aspect, the present invention provides a process for the preparation of a compound having the formula

NH

2

R

1

R

2 N R() 15 wherein R 1 is phenyl or thienyl, each of which is unsubstituted or substituted with one or two substituents independently selected from alkyl, alkoxy or halo; R 2 is lower alkyl, cycloalkyl, phenyl or phenyl alkyl wherein each of said phenyl is unsubstituted or substituted with one or two substituents independently selected from alkyl, alkoxy or halo; and R 3 is phenyl, pyridyl or thienyl, each of which is unsubstituted or substituted with one or two substituents selected from alkoxy or di(loweralkyl)amino; i 20 the process comprising reacting an aryl, heteroaryl, or a heterocyclic bromide having the formula R 3 -Br wherein R 3 is as defined above with a carboxylic acid derivative having the formula R 2

-CH

2 -CO-Y, wherein Y is OH or CI, and R 2 is as defined above, with N,O-dimethylhydroxylamine hydrochloride, 1-(3dimethylaminopropyl)-3-ethylcarbodiimide or 1-hydroxybenzotriazole hydrate and triethylamine, and S 25 isolating a first intermediate compound having the formula R 3

-CO-CH

2

-R

2 reacting the first intermediate compound having the formula R 3

-CO-CH

2

-R

2 with an aldehyde having the formula R 1 -CHO, wherein R 1 is as defined above, and malononitrile in the presence of an ammonium salt under anhydrous conditions, and isolating a second intermediate compound having the formula

R

1 NC R 2

H

2 N N R 3 reacting the second intermediate compound with formamide at reflux for from about 1 to about 24h, and isolating the compound of formula I.

In yet another aspect, the present invention provides a process for the preparation of o j compounds having the formula LibC/475183speci

NH

2

R'

R6 N N R(I) wherein R 1 is phenyl or thienyl, each of which is unsubstituted or substituted with one or two substituents independently selected from alkyl, alkoxy or halo; R 2 is lower alkyl, cycloalkyl, phenyl or phenyl alkyl wherein each of said phenyl is unsubstituted or substituted with one or two substituents independently selected from alkyl, alkoxy or halo; and R 3 is phenyl, pyridyl or thienyl, each of which is unsubstituted or substituted with one or two substituents selected from alkoxy or di(loweralkyl)amino; the process comprising reacting an aryl, heteroaryl, or a heterocyclic bromide having the formula R 3 -Br with a carboxylic acid derivative having the formula R 2

-CH

2 -CO-Y, wherein Y is OH or CI, and R 2 is as defined above, with N,O-dimethylhydroxylamine hydrochloride, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide or 1hydroxybenzotriazole hydrate and triethylamine, and isolating a first intermediate compound having the formula R3-CO-CH 2 -R2; treating the first intermediate compound having the formula R 3

-CO-CH

2

-R

2 with a compound having the formula NC N, wherein R 1 is as described above, at reflux in an alcoholic solvent, and isolating a second intermediate product having the formula NC R 2

H

2 N N R and reacting the second intermediate compound with formamide at reflux for from about 1 to about 24h, and isolating the desired product.

In still another aspect, the present invention provides a process for the preparation of S 20 compounds having the formula

NH

2

R

1 *0 3 N N R (I) wherein R 1 is phenyl or thienyl, each of which is unsubstituted or substituted with one or two substituents independently selected from alkyl, alkoxy or halo; R 2 is lower alkyl, cycloalkyl, phenyl or phenyl alkyl wherein each of said phenyl is unsubstituted or substituted with one or two substituents independently selected from alkyl, alkoxy or halo; and R 3 is phenyl, pyridyl or thienyl, each of which is unsubstituted or substituted with one or two substituents selected from alkoxy or di(loweralkyl)amino; the process comprising reacting an aryl, heteroaryl, or a heterocyclic bromide having the formula R 3 -Br with a carboxylic acid derivative having the formula R 2

-CH

2 -CO-Y, wherein Y is OH or Cl, and R 4 is as defined above, S 3o0 ith N,0-dimethylhydroxylamine hydrochloride, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide or 1- LibC/475183speci hydroxybenzotriazole hydrate and triethylamine, and isolating a first intermediate compound having the formula R 3

-CO-CH

2

-R

2 treating the first intermediate compound having the formula R 3

-CO-CH

2

-R

2 with a compound

R

1 having the formula NC CN, wherein R 1 is as described above, at reflux in an alcoholic solvent, and isolating a second intermediate product having the formula

R

1 NC R 2

H

2 N N R3; and reacting the second intermediate compound with sulfuric acid and heating followed by treatment with triethyl orthoformate at reflux for from about 1 to about 24h, and isolating a third intermediate compound having the structure OH R' R2

*N

N N R treating the third intermediate compound with a chlorinating agent, and isolating a fourth intermediate product having the formula Cl R 1 NN N R treating the fourth intermediate compound with ammonia, and isolating the compound of formula I.

15 Detailed Description of the Invention The present invention relates to novel 5,6,7-trisubstituted-4-aminopyrido[2,3-d]pyrimidine compounds of Formula above that are useful in inhibiting adenosine kinase, a method of inhibiting adenosine kinase with such compounds, to pharmaceutical compositions containing such compounds, 0* to a method of using such compounds for inhibiting adenosine kinase, and to novel 5,6,7trisubstituted-4-aminopyrido[2,3-d]pyrimidine compounds.

The present invention relates to a compound of formula I as described above.

In a more preferred embodiment of the present invention is a compound of Formula above, wherein R 3 is selected from the group consisting of: 4-(dimethylamino)phenyl; 5-dimethylamino-2pyridinyl; 5-methoxy-2-pyridinyl; 4-methoxyphenyl; and thiophen-2-yl.

In a more preferred embodiment of the present invention is a compound of Formula above, wherein R 2 is selected from the group consisting of: ethyl; 3-fluorophenyl; 3-fluoro-4-methylphenyl; 3,4-dimethoxyphenyl; 3-methoxyphenyl; 4-methoxyphenyl; pentyl; phenyl; 3-(2-propyl)phenyl; and 4- (2-propyl)phenyl.

The present invention relates to novel 5,6,7-trisubstituted-4-aminopyrido[2,3-d]pyrimidine FF3,,'compounds of Formula above that are useful in inhibiting adenosine kinase, a method of inhibiting LibC/475183speci adenosine kinase with such compounds, to pharmaceutical compositions containing such compounds, to a method of using such compounds for inhibiting adenosine kinase, and to novel 5,6,7trisubstituted-4-aminopyrido2,3-dlpyrimidime compounds.

The present invention relates to a compound of formula I as described above.

In a more preferred embodiment of the present invention is a compound of Formula above, wherein R 3 is selected from the group consisting of: 4-(dimethylamino)phenyl; 5-dimethylamino-2pyridinyl; 5-methoxy-2-pyridinyl; 4-methoxyphenyl; and thiophen-2-yI.

In a more preferred embodiment of the present invention is a compound of Formula above, wherein R 2 is selected from the group consisting of: ethyl; 3-fluorophenyl; 3-fluoro-4-methylphenyl; 3,4-dimethoxyphenyl; 3-methoxyphenyl; 4-methoxyphenyl; pentyl; phenyl; 3-(2-propyl)phenyl; and 4- (2-propyl)phenyl.

In another more preferred embodiment of the present invention is a compound of Formula (I) above, wherein RI is selected from the group consisting of: 3-bromophenyl; 3-bromo-4-fluorophenyl; 4-bromothiophen-2-yl; 3-chlorophenyl; 3,4-dimethoxyphenyl; 3-fluorophenyl; 3-fluoro-4-methylphenyl; and 4-(2-propyl)phenyl.

Exemplary and preferred compounds of the invention include: 4-amino-5-(3-bromo-4-fluorophenyl)-6-pentyl-7-(thiophen-2-yl)pyrido[2,3-dpyrimidine; 4-amino-5-(3-bromophenyl)-6-(4-methoxyphenyl)-7-(4-methoxyphenyl)pyrido[2,3-d]pyrimidine; 4-amino-5-(3-bromophenyl)-6-ethyl-7-(thiophen-2-yl)pyrido[2,3-d]pyrimidine; 20 4-amino-5-(3-bromophenyl)-6-pentyl-7-(th iophen-2-yl) pyrido[2,3-d] pyri mid ine; 4-amino-5-(3-bromophenyl)-6-(3,4-dimethoxyphenyl)-7-(thiophen-2-y)pyrido[2,3-d]pyrimidine; 4-amino-5-(3-bromophenyl)-6-(4-(2-propyl)phenyl)-7-(4-(dimethylamino)phenyl)pyrido[2,3dipyrimidine; 4-amino-5-(3-bromophenyl)-6-(3-methoxyphenylmethyl)-7-(thiophen-2-yl)pyrido[2,3-d]pyrimidine; .25 4-amni no-5-(3-bromophenyl)-6-(3,4-d imethoxyphenyl)-7-(4-(d imethyl amino) phenyl) pyrido[2,3- 4-amnino-5-(4-bromoth iophen-2-yl)-6-(3,4-d imethoxyphenyl)-7-(th iophen-2-yl)pyrido[2,3-d] pyri mid m e; 4-amino-5-(3-chlorophenyl)-6-(3,4-dimethoxyphenyl)-7-(thiophen-2-yl)pyrido[2,3-d]pyrimidine; 4-amino-5-(3-chlorophenyl)-6-(3,4-dimethoxyphenyl)-7-(4-(dimethylamino)phenyl)pyrido[2,3d]pyrimidine; 4-amino-5-(4-bromothiophen-2-yl)-6-(3,4-dimethoxyphenyl)-7-(4-(dimethylamino)phenyl)pyrido[2,3d]pyrimidine; 4-amino-5-phenyl-6-(3,4-di methoxyphenyl)-7-(5-methoxy-2-pyrid inyl)pyrido [2,3-d]pyri mid ine; 354-amino-5-(3-chlorophenyl)-6-(3,4-dimethoxyphenyl)-5-(5-dmethoyam-2-pyridinyl)pyrido[2,3-yiiie d]pyrimidine; 4-amino-5,6-bis(4-(2-propyl)phenyl-7-(4-dimethylaminophenyl)pyrido[2,3-d]pyrimidine; 4-amino-5,6-diphenyl-7-(4-dimethylaminophenyl)pyrido [2,3-dipyrimidine; amino-5,6-bis(3-fluorophenyl)-7-(4-dimethylaminophenyl)pyrido[2,3-dlpyrimidine; 4-amino-5,6-bis(3,4-dimethoxyphenyl)-7-(4-dimethylaminophenyl)pyrido[2,3-d]pyrimidine;

ZI

LibC/4751 83speci 4-amino-5,6-bis(3-fluoro-4-methylphenyl)-7-(4-dimethylaminophenyl)pyrido[2,3-d]pyrimidine; 4-amino-5,6-bis(3-fluoro-4-methylphenyl)-7-(thiophen-2-yl)pyrido[2,3-d]pyrimidine; 4-amino-5,6-diphenyl-7-(thiophen-2-yl)pyrido [2,3-d]pyrimidine; 4-amino-5,6-diphenyl-7-(5-dimethylamino-2-pyridinyl)pyrido [2,3-d]pyrimidine; s 4-amino-5-phenyl-6-(3,4-dimethoxyphenyl)-7-(5-(dimethylamino)pyridin-2-yl)pyrido[2,3-d]pyrimidine; 4-amino-5-(3-chlorophenyl)-6-phenyl-7-(5-dimethylamino-2-pyridinyl)pyrido[2,3-d]pyrimidine; 4-amino-5-(4-bromothiophen-2-yl)-6-phenyl-7-(5-dimethylamino-2-pyridinyl)pyrido [2,3-d]pyrimidine; 4-amino-5-(3-bromophenyl)-6-phenyl-7-(6-dimethylamino-3-pyridinyl)pyrido[2,3-dpyrimidine; 4-amino-5-(4-bromothienyl)-6-cyclopropyl-7-(6-dimethyamino-3-pyridinyl)pyrido[2,3-d]pyrimidine; 4-amino-5-(3-bromophenyl)-6-cyclohexyl-7-(6-dimethylamino-3-pyridinyl)pyrido[2,3-d]pyrimidine; and 4-amino-5-(3-bromophenyl)-6-pentyl-7-(6-dimethylamino-3-pyridinyl)pyrido[2,3-d]pyrimidine; and pharmaceutically acceptable salts and amides thereof.

The term "cycloalkyl" refers to a cyclic saturated hydrocarbon radical having from 3 to 7 ring atoms. Examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl. Cycloalkyl is also described as C3-C8cycloalkyl.

The term "cycloalkyl-loweralkyl" refers to a loweralkyl radical as defined below substituted with a cycloalkyl group as defined above by replacement of one hydrogen atom. Examples of cycloalkylloweralkyl include cyclopropylmethyl, cyclobutylethyl, cyclopentylmethyl, cyclohexylmethyl and cycloheptybutyl, and the like.

The term "loweralkyl", as used herein, refers to saturated, straight- or branched-chain hydrocarbon radicals containing from one to six carbon atoms including, which may be unsubstituted or substituted by independent replacement of one, two or three of the hydrogen atoms thereon with Cl, Br, F, I, cyano, carboxamido, hydroxy, loweralkoxy, amino, loweralkylamino, di(loweralkylamino) or N-loweralkyl-N-loweralkoxyamino groups. Examples of loweralkyl include, but are not limited to, S 25 methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, hydroxyethyl, methoxymethyl, trifluoromethyl, 3-cyanopropyl, 3-carboxamidopropyl, and the like. In certain cases, the group "Ci- Calkyl" is described and has a similar meaning as above for loweralkyl but is more specifically *recited. Likewise, the term "Co-C6alkyl" indicates the carbon atoms which may be present in the alkyl chain including zero. These terms are also provided adjacent to aryl or heteroaryl or other generic group and represent or have the same meaning as, for example, "arylalkyl" or "heteroarylalkyl".

The term "loweralkoxy" refers to a loweralkyl radical which is appended to the molecule via an ether linkage through an oxygen atom), as for example methoxy, ethoxy, propoxy, 3-propoxy, 2methyl-2-propoxy, tert-butoxy, pentyloxy, hexyloxy, isomeric forms thereof and the like. This term is also described as Ci-Calkyloxy.

The term "mammal" has its ordinary meaning and includes human beings.

In a further aspect of the present invention pharmaceutical compositions are disclosed which comprise a compound of the present invention in combination with a pharmaceutically acceptable carrier.

The present invention includes one or more compounds, as set forth above, formulated into cempositions together with one or more non-toxic physiologically tolerable or acceptable diluents, /LibC/475183speci 9 carriers, adjuvants or vehicles that are collectively referred to herein as diluents, for parenteral injection, for oral administration in solid or liquid form, for rectal or topical administration, or the like.

As is well known in the art, a compound of the present invention can exist in a variety of forms including pharmaceutically-acceptable salts, amides and the like.

Compositions may be prepared that will deliver the correct amount of a compound or compounds of the invention. The following dosages are thought to provide the optimal therapy: iv infusions: 0.1- 250nmol/kg/minute, preferably from 1-50nmol/kg/minute; oral: 0.01-250Mol/ kg/day, preferably from about 0.1-50Mol/kg/day; these oral molar dosage ranges correspond to 0.005- 125mg/kg/day, preferably 0.05-25mg/kg/day. For treatment of acute disorders the preferred route of administration is intravenous; the preferred method of treating chronic disorders is orally by means of a tablet or sustained release formulation.

"Pharmaceutically-acceptable amide" refers to the pharmaceutically-acceptable, nontoxic amides of the compounds of the present invention which include amides formed with suitable organic acids or with amino acids, including short peptides consisting of from 1-6 amino acids joined by amide linkages which may be branched or linear, wherein the amino acids are selected independently from naturally-occurring amino acids, such as for example, glycine, alanine, leucine, valine, phenylalanine, proline, methionine, tryptophan, asparagine, aspartic acid, glutamic acid, glutamine, serine, threonine, .::lysine, arginine, tyrosine, histidine, omithine, and the like.

"Pharmaceutically-acceptable salts" refers to the pharmaceutically-acceptable, nontoxic, inorganic or organic acid addition salts of the compounds of the present invention, as described in greater detail below.

The compounds of the present invention can be used in the form of pharmaceuticallyacceptable salts derived from inorganic or organic acids. These salts include, but are not limited to, the following: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, 25 camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, S ethanesulfonate, flavianate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxy-ethanesulfonate, lactate, maleate, methanesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, palmate, petinate, persulfate, 3phenylpropionate, phosphate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, and undecanoate.

Appropriate cationic salts are also readily prepared by conventional procedures such as treating an acid of Formula I with an appropriate amount of base, such as an alkali or alkaline earth metal hydroxide, eg., sodium, potassium, lithium, calcium, or magnesium, or an organic base such as an amine, eg., dibenzylethylenediamine, cyclohexylamine, dicyclohexylamine, triethylamine, piperidine, pyrrolidine, benzylamine, and the like, or a quaternary ammonium hydroxide such as tetramethylammonium hydroxide and the like.

Also, the basic nitrogen-containing groups can be quaternised with such agents as loweralkyl halides, such as methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dialkyl sulfates; long jsfchain halides such as decyl, lauryl, myristyl, and stearyl chlorides, bromides and iodides; arylalkyl LibC/475183speci halides like benzyl and phenethyl bromides, and others. Water or oil-soluble or dispersible products are thereby obtained.

The salts of the present invention can be synthesised from the compounds of Formula I which contain a basic or acidic moiety by conventional methods, such as by reacting the free base or acid with stoichiometric amounts or with an excess of the desired salt forming inorganic acid or base in a suitable solvent or various combinations of solvents.

Further included within the scope of the present invention are pharmaceutical compositions comprising one or more of the compounds of formula prepared and formulated in combination with one or more non-toxic pharmaceutically acceptable carriers compositions, in the manner described below.

Compositions suitable for parenteral injection may comprise pharmaceutically acceptable sterile aqueous or nonaqueous solutions, dispersions, suspensions or emulsions and sterile powders for reconstitution into sterile injectable solutions or dispersions. Examples of suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, and the like), suitable mixtures thereof, vegetable oils (such as olive oil) and injectable organic esters such as ethyl oleate. Proper fluidity may be maintained, for example, by *diesn sthe use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

These compositions may also contain adjuvants such as preserving, wetting, emulsifying, and dispersing agents. Prevention of the action of microorganisms may be ensured by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, and the like. It may also be desirable to include isotonic agents, for example, sugars, sodium chloride and the like. Prolonged absorption of the injectable pharmaceutical form may be brought about by the use of agents delaying absorption, for example, aluminium monostearate and gelatin.

If desired, and for more effective distribution, the compounds may be incorporated into slowrelease or targeted-delivery systems, such as polymer matrices, liposomes, and microspheres. They may be sterilised, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilising agents in the form of sterile solid compositions, which may be dissolved in sterile water, or some other sterile injectable medium immediately before use.

Solid dosage forms for oral administration may include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is admixed with at least one inert customary excipient (or carrier), such as sodium citrate or dicalcium phosphate, and additionally (a) fillers or extenders, as for example, starches, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, as for example, carboxymethylcellulose, alginates, gelatin. polyvinylpyrrolidone, sucrose and acacia; humectants, as for example, glycerol; disintegrating agents, as for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates and sodium carbonate; solution retarders, as for example paraffin; absorption accelerators, as for example, quaternary ammonium compounds; wetting agents, as for example, cetyl alcohol and glycerol monostearate; adsorbents, as for example, kaolin and bentonite; and lubricants, as for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate or O /I LibC/475183speci

NZ(/Z

11 mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules, using such excipients as lactose or milk sugar, as well as high molecular weight polyethylene glycols, and the like.

Solid dosage forms such as tablets, dragees, capsules, pills and granules may be prepared with coatings and shells, such as enteric coatings and others well known in this art. They may contain pacifying agents, and may also be of such composition that they release the active compound or compounds in a certain part of the intestinal tract in a delayed manner. Examples of embedding compositions which may be used are polymeric substances and waxes.

The active compounds may also be in micro-encapsulated form, if appropriate, with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilising agents and emulsifiers, as for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, in particular, cottonseed oil, groundnut oil, corn germ oil, olive oil, castor oil and sesame oil, glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan or mixtures of these substances, and the like.

Besides such inert diluents, these liquid dosage forms may also include adjuvants, such as wetting agents, emulsifying and suspending agents, sweetening, flavouring and perfuming agents.

~Suspensions, in addition to the active compounds, may contain suspending agents, as for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminium metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances, and the like.

Compositions for rectal or vaginal administrations are preferably suppositories which can be :prepared by mixing the compounds of this invention with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax, which are solid at ordinary temperatures but liquid at body temperature and therefore, melt in the rectum or vaginal cavity and release the active component.

Dosage forms for topical or transdermal administration of a compound of this invention further include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or transdermal patches. Transdermal administration via a transdermal patch is a particularly effective and preferred dosage form of the present invention. The active component is admixed under sterile conditions with a pharmaceutically acceptable carrier and any needed preservative, buffers or propellants as may be required. It is known that some agents may require special handling in the preparation of transdermal patch formulations. For example, compounds that are volatile in nature may require admixture with special formulating agents or with special packaging materials to assure proper dosage delivery. In 746, addition, compounds which are very rapidly absorbed through the skin may require formulation with j LibCI475183speci absorption-retarding agents or barriers. Ophthalmic formulations, eye ointments, powders and solutions are also contemplated as being within the scope of this invention.

The present compounds may also be administered in the form of liposomes. As is known in the art, liposomes are generally derived from phospholipids or other lipid substances. Liposomes are formed by mono- or multi-lamellar hydrated liquid crystals that are dispersed in an aqueous medium.

Any non-toxic, physiologically acceptable and metabolisable lipid capable of forming liposomes may be used. The present compositions in liposome form may contain, in addition to the compounds of the present invention, stabilisers, preservatives, excipients, and the like. The preferred lipids are the phospholipids and the phosphatidyl cholines (lecithins), both natural and synthetic.

Methods to form liposomes are known in the art. See, for example, Prescott, Ed., Methods in Cell Biology, Volume XIV, Academic Press, New York, N. (1976), p 33 et seq.

Synthetic Methods The compounds and processes of the present invention will be better understood in connection with the following synthetic schemes which illustrate the methods by which the compounds of the invention may be prepared. The R 1

R

2 and R 3 groups are as defined above unless otherwise noted below.

The compounds of the present invention may be synthesised by methods illustrated in Schemes 1-3.

Scheme 1 R 2 20 R 2

-CH

2 -CO-Y CH 3

-NH-O-CH

3

-R

2

-CH

2

-CO-N(CH

3

)-O-CH

3

R

3 -Br R 3 1 2 3 4

SR

1 NH R' NC R 2 0 R 2 3 R2 0 3 H NH 2 O O 3 R R 1 -CO-H NC-CH 2 -CN -H 2 N N R N N R 3 5 6 7 8 9 (I) In accordance with Scheme 1, the 5,6,7-trisubstituted compounds wherein R 3 is phenyl, pyridyl or thienyl, R 1 is phenyl or thienyl, and R 2 is loweralkyl, phenyl, or phenylalkyl are prepared by a modification of a method of Kambe et al., Synthesis, 1980, 366-368.

N-methoxy-N-methylamide compounds may be prepared from the appropriate carboxylic acid derivative the "R 2 reagent"), wherein Y is OH or CI, and R 2 is loweralkyl, phenyl or phenylalkyl, by treatment with N,0-dimethylhydroxylamine hydrochloride and 1-(3-dimethylaminopropyl)-3ethylcarbodiimide (EDCI), t-butanol, and triethylamine. The reaction may be performed in methylene chloride, or a similar suitable solvent, such as for example, toluene or THF, at ambient temperature for about 8h to about 24h.

Compound is then reacted with compounds the "R3 reagent") wherein R 3 is substituted phenyl, pyridyl, or thienyl to prepare compound according to the literature procedure of Nahm and Weinreb (Tetrahedron Lett. 1981, 22: 3815). Compounds are obtainable commercially or easily 2 LibC/475183speci prepared by standard methods in the art. Compound is then reacted with an appropriately substituted aldehyde the "R 1 Reagent"), wherein R 1 is phenyl or thienyl, and malononitrile by heating in the presence of ammonium acetate, or another suitable ammonium salt, such as for example, ammonium propionate, ammonium iodide, or the like, in a suitable solvent to produce compound Suitable solvents include ethanol, benzene, toluene, methylene chloride, DMF, THF, dioxane, and the like. The water of the reaction may removed by use of a Dean Stark apparatus or by another suitable means, such as 4 A molecular sieves. The reaction may be performed at from about 0 C to about 200°C, and preferably at the reflux temperature of the solvent, for from about 1h to about 24h, preferably about 4h to 8h. The product is preferably purified by chromatography after isolation from the reaction mixture.

The appropriate aldehyde starting materials may be obtained commercially, or may be prepared easily, for example by reduction of esters or acids with DIBAL or another suitable hydride reducing agent, or oxidation of alcohols under Swern conditions, for example. Aliphatic aldehydes do not work effectively by this route. The ketone may, however, include R 5 as alkyl groups.

Compound is then treated with excess formamide by heating at reflux. The formation of product is monitored by TLC, and when the reaction is complete (after about 1 to about 8h) the reaction mixture is cooled to room temperature. The desired 5,6,7-trisubstituted pyrido[2,3d]pyrimidine product is then removed by filtration and purified by column chromatography.

Stereoisomers produced in the reduction process or step(s) are included within the scope of the invention. The stereoisomers may be isolated and purified by conventional means.

In an alternate procedure, compound is treated by heating with formamidine acetate in ethoxyethanol or diglyme, followed by purification by flash chromatography. In another alternate procedure, compound and ammonium sulfate are heated at reflux in triethyl orthoformate for about 1 to about 8h, but preferably about 2h. The reaction mixture is cooled and added to a mixture of ammonia in ethanol. The mixture is stirred for about 12 to 24h at 25 0 C, then at reflux for from one to 4h, and the solvent is removed in vacuo. The residue is purified by trituration with chloroform/ethyl acetate, and the product may be converted to a hydrochloride salt by suspension in 2M HCI, followed by lyophilisation.

Scheme 2 R2 NC R2 R 2^ 1 R 3 0 3

R

3

H

2 N N R 3 10 8 Scheme 2 illustrates an alternate method for preparing the compounds of Scheme 1.

Compounds prepared as described above, are reacted with a dicyanoalkene compound (10) by heating at reflux in an alcohol solvent, for example, ethanol or n-butanol to give the compound The dicyano compounds (10) may be prepared from the precursor aldehyde by treatment with IS7 alononitrile in 1:1 H20:EtOH in the presence of a catalytic amount of glycine according to the it/od of Bastus (Tetrahedron Lett., 1963: 955).

OFF\ LibC/475183speci Scheme 3

NH

2

NH

2

R'

(0N R 3H R1-R2 NH 2 K- (I) R-=-R2 N NH2 N N H 2 R3 11 12 13 14 Scheme 3 illustrates an alternate method for preparing compounds of Formula wherein R 2 and R 1 are the same substituent. A bis-substituted acetylene derivative (11) is treated with catecholborane in THF at reflux for from about 8 to about 48h, then 4,6-diamino-5-iodo-pyrimidine saturated aqueous sodium bicarbonate, 3N aqueous sodium hydroxide, and tetrakis(triphenylphosphine)palladium(0) are added. The mixture is then heated at reflux for from about 8 to about 48h to give the substituted pyrimidine compound Compound (13) is then treated with the appropriately substituted aldehyde compound (14) to give the desired compound of formula Scheme 4 0 r R2 N N 5, cf Scheme 2 R" 20 NR'R" 0 0 0 2 2 R 2 15 X- N O-Et X N 0 X N 18 X R2 616 17 procedure is particularly useful when it is desired to have a substituted phenyl, pyridyl or thienyl moiety in the R2 position.

A compound (15) containing the desired R2 moiety may be reacted with an acyl halide of a halo-substituted compound the ring of which for purposes of illustration only, is shown as a pyridyl ring 2-halo-5-pyridine carboxylic acid halide), to give the compound Compound (17) in turn i heated to decarboxylate and give compound (18).

in turn is heated to decarboxylate and give compound (18).

LibC/475183speci Alternatively, compound (16) may be treated in a two-step procedure, first with Nmethoxymethylamine HCI, then treating the intermediate with compound (19) under Grignard conditions to prepare compound (18).

Compound (18) may then be reacted with an appropriate amine compound where compound (20) may be a heterocyclic compound, such as piperidine, pyrrolidine, or morpholine, for example, or may be a protected or substituted amine, ie. wherein R' and R" are either substituents or amine-protecting groups, or R' and R" are taken together with the N atom to which they are attached to form a heterocyclic ring, in order to prepare compound Method of Inhibiting Kinase In yet another aspect of the present invention a method of inhibiting adenosine kinase is disclosed. In accordance with that process, an adenosine kinase enzyme is exposed to an effective inhibiting amount of an adenosine kinase inhibitor compound of the present invention. Preferred such compounds for use in the method are the same as set forth above. Means for determining an effective inhibiting amount are well known in the art.

The adenosine kinase to be inhibited can be located in vitro, in situ or in vivo. Where the adenosine kinase is located in vitro, adenosine kinase is contacted with the inhibitor compound, typically by adding the compound to an aqueous solution containing the enzyme, radiolabelled substrate adenosine, magnesium chloride and ATP. The enzyme can exist in intact cells or in isolated subcellular fractions containing the enzyme. The enzyme is then maintained in the presence of the inhibitor for a period of time and under suitable physiological conditions. Means for determining maintenance times are well known in the art and depend inter alia on the concentrations of enzyme and the physiological conditions. Suitable physiological conditions are those necessary to maintain adenosine kinase viability and include temperature, acidity, tonicity and the like. Inhibition of adenosine kinase can be performed, by example, according to standard procedures well known in the 25 art (Yamada, et al., Comp. Biochem. Physiol. 1982, 71B: 367-372).

Where the adenosine kinase is located in situ or in vivo, a compound of the invention is typically administered to a fluid perfusing the tissue containing the enzyme. That fluid can be a naturally occurring fluid such as blood or plasma or an artificial fluid such as saline, Ringer's solution and the like. A method of inhibiting adenosine kinase in vivo is particularly useful in mammals such as humans. Administering an inhibitor compound is typically accomplished by the parenteral (eg., intravenous injection or oral) administration of the compound. The amount administered is an effective inhibiting or therapeutic amount.

By a "therapeutically-effective amount" of the compound of the invention is meant a sufficient amount of the compound to treat or mitigate adenosine kinase related disorders which broadly include those diseases, disorders, or conditions which are benefited by inhibition of adenosine kinase, at a reasonable benefit/risk ratio applicable to any medical treatment. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention is to be decided by the attending physician within the scope of sound medical judgment. The specific therapeutically- Seffective dose level for any particular patient will depend upon a variety of factors including the 'disorder being treated and the severity of the disorder, activity of the specific compound employed; F/ LibC/47583speci OF -\C the specific composition employed; the age, body weight, general health, gender and diet of the patient; the time of administration, route of administration, and rate of excretion of the specific compound employed: the duration of the treatment; drugs used in combination or coincidental with specific compound employed; and the like factors well known in the medical arts and well within the capabilities of attending physicians.

Compounds of the present invention inhibit adenosine kinase activity in vitro and in vivo. In vitro adenosine kinase activity can be measured using any of the standard procedures well known in the art. By way of example, cells containing adenosine kinase, such as IMR-32 human neuroblastoma cells, are cultured in the presence and absence of an inhibitor. Inhibition is measured lo as the ability to inhibit phosphorylation of endogenous or externally applied 1 4 C-adenosine by these cells. The cells can be intact or broken. The specificity of adenosine kinase inhibitory activity is determined by studying the effects of inhibitors on adenosine Al and A2cc receptor binding, adenosine deaminase activity and adenosine transport.

Compounds of the present invention are effective in inhibiting adenosine kinase activity in vivo.

Numerous animal models for studying adenosine kinase activity and the affects of inhibiting such activity are well known in the art. By way of example, adenosine kinase inhibitors have been reported i to protect rodents mice and rats) from seizures induced by the subcutaneous administration of pentylenetetrazol (PTZ). Typically the rodents are injected with various doses of a given inhibitor followed at various times by the subcutaneous administration of from about 10 to about 500mg per kilogram of PTZ. The injected animals are then observed for the onset of seizures.

The compounds of the invention were tested in vivo in the hot plate test of analgesia in mammals such as mice. For example, the compounds of examples 19 and 27 in the procedure described directly below were tested thirty minutes after pretreatment with the drugs (30.mol/kg i.p.) for latency to 10' jump (in seconds). The longer the number of seconds, the more effective the drug at masking the pain felt from the hot plate. Compound 19 resulted in 142.13s relative to the vehicle alone of 72.76±10.51s. Compound 27 resulted in 154.86s. Compounds of the invention are therefore potent pain relievers as demonstrated in this animal model.

Mouse Hot Plate Assay Male CF1 mice (Charles River) of approximately 25-30g body weight are pretreated with 10mUkg of the test compounds, i.p. or p.o, in groups of 8 animals per dose. At the end of the pretreatment period, the mice are placed in an Omnitech Electronics Automated 16 Animal Hot Plate Analgesia Monitor (Columbus, OH; Model AHPI6AN) in individual, 9.8 x 7.2 x 15.3cm (1 x w x h) plastic enclosures on top of a copper plate warmed to 55 0 C. infrared sensors located near the top of each enclosure record beam crossings that occur as the mice jump off of the heated surface. Latency times for each jump are automatically recorded, and latency to both the first and tenth jumps are used for data analysis. Mice that do not reach the criteria of 10 jumps by 180s are immediately removed from the hotplate to avoid tissue damage, and they are assigned the maximum value of 180s as their latency to tenth jump.

LibC/475183sped Numerous other animal models of adenosine kinase activity have been described [See, eg..

Davies,, et al., Biochem. Pharmacol., 33:347-355 (1984); Keil, et al., Eur. J. PharmacoL., 271:37-46 (1994); Murray, et al., Drug Development Res., 28:410-415 (1993)].

Numerous inhibitor compounds of the present invention were tested in vitro and found to inhibit adenosine kinase activity. The results of some representative studies are shown in Table 1 below.

The data indicate that the compounds inhibit adenosine kinase.

Table 1 Inhibition of Adenosine Kinase by Representative Compounds of the Invention ft 10 2

C

2 1 f 20

C

C

C•

Compound of Example No. ICso (nM) 1 4 3 6 1 9 23 11 0.3 12 1 4 27 9 39 40 3 41 2 Method of Treating Cerebral Ischaemia, Epilepsy, Nociperception (Nocieption) (Pain), Inflammation including conditions such as Septic Shock due to Sepsis Infection In yet another aspect of the present invention a method of treating cerebral ischaemia, epilepsy, nociperception or nociception, inflammation including conditions such as septic shock due to sepsis infection in a human or lower mammal is disclosed, comprising ad'ministering to the mammal a therapeutically effective amount of a compound.

Alterations in cellular adenosine kinase activity have been observed in certain disorders.

Adenosine kinase activity was found to be decreased, relative to normal liver, in a variety of rat hepatomas: activity of the enzyme giving a negative correlation with tumour growth rate (Jackson, et Br. J. Cancer, 1978. 37: 701-713). Adenosine kinase activity was also diminished in regenerating liver after partial hepatectomy in experimental animals (Jackson, et al., Br. J. Cancer, 1978, 37: 701- 713). Erythrocyte Adenosine kinase activity was found to be diminished in patients with gout (Nishizawa, et al, Clin. Chim. Acta 1976, 67: 15-20). Lymphocyte adenosine kinase activity was decreased in patients infected with the human immunodeficiency virus (HIV) exhibiting symptoms of AIDS, and increased in asymptomatic HIV-seropositive and HIV-seronegative high-risk subjects, compared to normal healthy controls (Renouf, et al., Clin. Chem. 1989, 35: 1478-1481). It has been suggested that measurement of adenosine kinase activity may prove useful in monitoring the clinical progress of patients with HIV infection (Renouf, et al., Clin. Chem. 1989, 35: 1478-1481). Sepsis infection may lead to a systemic inflammatory syndrome (SIRS), characterised by an increase in cytokine production, neutrophil accumulation, homodynamic effects, and tissue damage or death.

The ability of adenosine kinase inhibitor to elevate adenosine levels in tissues has been demonstrated to ameliorate syndrome symptoms, due to the known anti-inflammatory effects of adenosine.

irestein, etal., J. of Immunology, 1994, pp. 5853-5859). The ability of adenosine kinase inhibitors to elyate adenosine levels is expected to alleviate pain states, since it has been demonstrated that LibC/475183spec administration of adenosine or its analogues results in antinociception or antinociperception.

(Swaynok, et al., Neuroscience, 1989, 32: No. 3, pp. 557-569).

The following Examples illustrate preferred embodiments of the present invention and are not limiting of the specification and claims in any way.

Example 1 4-amino-5-(3-bromo-4-fluorophenyl)-6-pentyl 7-(4-(dimethylamino)phenyl)pyrido[2,3-d]pyrimidine 4-(3-Bromo-4-fluorophenyl)-3-cyano-6-(dimethylaminophenyl)-5-pentyl-2-pyridineamine (951mg, 1.98mmol) was suspended in 2-ethoxyethanol followed by addition of formamidine acetate (411mg, 3.95mmol). The reaction was heated to 130°C for two days during which additional formamidine acetate (2-3eq each) was added at several hour intervals. After this time the reaction was cooled, concentrated in vacuo, and the residue was triturated with CH 2 CI2 and filtered. The filtrate was purified by flash chromatography MeOH/CH2CI2) which gave a red oil that was triturated with ethyl ether to yield the title compound as a yellow solid (174mg, MS 508/510 IR (cm-1) 3480, 2920, 1610, 1550, 820.

The 4-(3-bromo-4-fluorophenyl)-3-cyano-6-(dimethylaminophenyl)-5-pentyl-2-pyridineamine was prepared as follows: I a. 1-(4-dimethylaminophenyl)heptan--one Triethylamine (19.6g. 194mmol) was added dropwise to a suspension of N,Odimethylhydroxylamine hydrochloride (6.93g, 71mmol) in anhydrous CH 2

CI

2 at 0°C.

20 Heptanoylchloride (9.60g, 65mmol) was then added dropwise and the reaction was stirred 1h. The crude product mixture was poured into water and the separated aqueous phase was extracted with a CH 2

CI

2 The combined organic layers were washed with aq. HCI, sat. NaHCO 3 brine, dried (Na2SO4), and concentrated in vacuo to give 10g N-methyl-N-methoxyheptanamide as a yellow oil.

n-Butyllithium (2.5M in hexanes, 51mL, 127mmol) was added dropwise to 4-bromo-N,N- 25 dimethylaniline (23.1g, 115mmol, Aldrich Chemical Co.) in anhydrous THF at -78 0 C. After 10min. a solution of N-methyl-N-methoxyheptanamide (10.0g, 57.7mmol) in 20mL THF was added dropwise via s cannula. The reaction was allowed to proceed 1h, then quenched with 1N aq. HCI and carefully poured into sat NaHCO 3 The aqueous layer was extracted with ethyl ether, and the combined organic fraction was washed with water, brine, dried (MgSO4), and concentrated in vacuo. Flash chromatography (15% EtOAc/hexanes) yielded 1-(4-dimethylaminophenyl)heptan-1-one as a yellow solid (6.49 g, MS 234 ib. 4-(3-bromo-4-fluorophenyl)-3-cyano-6-(dimethylaminophenyl)-5-pentyl-2-pyridineamine 1-(4-Dimethylaminophenyl)heptan-1-one (2.15g, 9.21mmol), 3-bromo-4-fluorobenzaldehyde (1.87g, 9.21mmol, the R3 reagent), malononitrile (0.91g, 13.8mmol), and NH 4 0Ac (1.42g, 18.4mmol) were dissolved in benzene (75mL) and heated to reflux.

After three days the crude reaction mixture was partitioned between EtOAc and H 2 0. The organic layer was washed with brine, dried over Na 2

SO

4 and concentrated in vacuo. The residue was triturated with Et20, and the resulting solid was collected by filtration yielding 1.25g of the desired product as a yellow solid (28 MS 444/446 LibC/475183speci Examples 2-10 Following the procedures of Example 1, except substituting for R 3

R

2 and R 1 as indicated in Table 2 below, optionally omitting compounds of Examples 2-10 were prepared as described in Table Table 2 the appropriate reagents required the step of preparing the HCI salt, 3 below.

0000 0 see.

so 0 a 0 Examples 2-9 Ex No. IR 3 Reagent (for 7-position) R 2 Reagent (for 6-position) R 1 Reagent (for 2 2-bromothiophene heptanoyl chloride 3-bromo-4-fluorobenzaldehyde 3 1 -bromo-4-methoxybenzene 4-methoxyphenylacetyl chloride 3-bromobenzaidehyde 4 2-bromothiophene butanoyl chloride 3-bromobenzaidehyde .2-bromothiophene heptanoylchloride 3-bromobenzaldehyde 6 12-bromothiophene 3,4-dimethoxyphenylacetic acid 3-bromobenzaldehyde 7 1 -bromo-4- 4-(2-propyl)phenylacetic acid 3-bromobenzaldehyde (dimethylamino)benzene 8 2-bromothiophene 3-(3-methoxyphenyl)propionic acid 3-brombenzaldehyde 1 -bromo-4- 3,4-dimethoxyphenylacetic acid 3-bromobenzaldehyde (dimethylamino)benzene Table 3 Examples 2-9 Ex. No.jName Analytical Data 2 4-amino-5-(3-bromo-4-fluorophenyl)-6-pentyl-7-(thiophen-2- I R: 3480, 2920, 1610, 1550, 820; MS -yl)pyrido[2,3-djpyrimnidine m/z 508/51 3 4-amino-5-(3-bromophenyl)-6-(4-methoxyphenyl)-7-(4- IR: 3480, 3400, 3070,1610, 1550; MVS methoxyphenyl)pyrido[2,3-dlpyrimidine m/z 51 3&51 5 4 4-amino-5-(3-bromophenyl)-6-ethyl-7-(thiophen-2- IR: 3470, 3390,3060, 1550, 1425; MVS ____l)pyrido[2,3-dlpyrimidine m/z 411 &413 4-amino-5-(3-bromophenyl)-6-pentyl-7-(thiophen-2- IR: 3480, 3300, 3040, 1550, 1420; MS yl)pyrido[2,3-dlpyrimidine rn/s 453&455 6 4-amino-5-(3-bromophenyl)-6-(3,4-dimethoxyphenyl)-7- IR: 3480, 3390, 3060, 1545, 1510; MS (thiophen-2-yl)pyrido[2,3-d]pyrimidine mlz 51 9&52 1 7 4-amino-5-(3-bromophenyl)-6-(4-(2-propyl)phenyl)-7-(4- IR: 3470, 3280, 3060, 1605,1540; MVS (dimethylamino)phenyl)pyrido[2,3-d]pyrimidine mlz 538&540 8 4-amino-5-(3-bromophenyl)-6-(3-methoxyphenylmethyl)-7- IR: 3440, 3040, 1635, 1600,1580; MS -(thiophen-2-yl)pyrido[2,3-d]pyrimidine hydrochloride mlz 503&505 9 4-amino-5-(3-bromophenyl)-6-(3,4-dimethoxyphenyl)-7-(4- I R: 3430, 3020, 1635, 1600, 1580; MS (dimethylamino)phenyl)pyrido[2,3-d]pyrimidine mlz 556&558 ,dihydrochioride Example 4-amino-5-(4-bromothiophen-2-yI)-6-(3,4-dimethoxyphenyl).7-(thiophen-2yl)pyrido[2,3-d]pyrimidine hydrochloride 3-Cyano-4-(4-bromothiophen-2-yl)-5-pentyl-6-(thiophen-2-yl)-2-pyridineamine (750mg, and formamidine acetate (312mg, 3.O0mmol) were taken up in l0mL- diglyme and heated to 15500. Additional formamidine acetate (leq) was added at 90 minute intervals over a total of 6h, then heating was continued overnight. The cooled reaction mixture was then partitioned between EtOAc and H20. The organic layer was washed with brine, dried over Na 2

SO

4 and concentrated in vacuo. Flash chromatography MeOH/CH 2

CI

2 gave a brown residue which was dissolved in a small amount of CH2CI 2 followed by addition of Et2O to precipitate the product (209mg, This p;T7 1 ,,mateial was converted to the hydrochloride salt using 7M ethanolic HCI followed by precipitation with n rp:lvv-:LibC/4751 B3speci Et 2 O and filtration of the product. IR: 525/527, 3420, 2930, 1580, 1510, 820 cm- 1 MS m/z 498 The 3-cyano-4-(4-bromothiophen-2-yl)-5-(3,4-dimethoxyphenyl)-6-(thiophen-2-yl)-2-pyridineamine was prepared as follows: 11 Ia. 2-(3,4-dimethoxyphenyl)-1 -(thien-2-yI)ethanone (3,4-Dimethoxyphenyl)acetic acid (13.0g, 66.4mmol) was suspended in anhydrous CH 2

CI

2 followed by addition of EDCI (15.3g, 79.7mmol), HOBt (20.6g, l52mmoI), triethylamine (8.06g, 79.7mmol), and N,O-dimethylhydroxylamine hydrochloride (6.48g, 66.4mmol). The reaction was stirred 3 days at ambient temperature after which the solvent was evaporated at reduced pressure.

The residue was partitioned between EtOAc and water. The organic layer was washed with aq HCI, sat NaHCO 3 brine, dried (Na2SO4), and concentrated in vacuo to give 10.5g of N-methyl-Nmethoxy-(3,4-dimethoxyphenyl)acetamide as a pale brown oil.

2-Lithiothiophene (1 .OM in THE, 33.OmL, 33.Ommol, Aldrich Chemical Co.) was added dropwise to N-methyl-N-methoxy-(3,4-dimethoxyphenyl)acetamide (5.26g. 22.Ommol) in anhydrous THF at ~15 -780C. The reaction was allowed to proceed 90mmn., then diluted with lO0mL Et 2 O and poured into 1N aq HCl The aqueous phase was extracted with Et2O and the combined organic fraction was 0. 0% 0 washed with brine, dried (Na2SO4), and concentrated in vacuo. Flash chromatography 000000 EtOAc/hexanes) yielded 2.91g 2-(3,4-dimethoxyphenyl)-1-(thien-2-yl)ethanone as a brown oil.

0 00 00 MS 263 280 (M+NH 4 20 11 b. 4-bromo-2-(2,2-dicyanoethenyl)thiophene 0 4-Bromo-2-thiophenecarboxaldehyde (6.92g. 36.2mmol) and malononitrile (2.39g, 36.2mmol) 0 0 0 0 0were dissolved in l00mL- 1:1 EtOH:H 2 0. A small spatula of glycine was added and the reaction was stirred at ambient temperature for 30mmn. The precipitated product was collected by suction filtration, 25washed with water, and dried under vacuum overnight. The result was 8.38g 4-bromo-2-(2,2dicyanoethenyl)thiophene as a light green solid. MS 238/240 11 c. 3-cyano-4-(4-bromothiophen-2-yI)-5-(3,4-dimethoxyphenyl)-6-(thiophen.2.yl)-2.pyridineamine o and heated to reflux. After 24h, the reaction mixture was cooled, diluted with EtOAc, and washed with water, brine, dried over Na2SO4, and concentrated in vacuo. Flash chromatography EtOAc/hexanes) gave the desired product (0.76 g, 26%) as a dark yellow solid.

Examples 11-18 Following the procedures of Example 10, except substituting the appropriate reagents required for R 3

R

2 and R 1 as indicated in Table 4 below, compounds of Examples 11-18 were prepared as described in Table 5 below.

Table 4 Examples 11-18 Ex. No. R 3 Reagent (for 7-position) R 2 Reagent (for 6-position) R 1 Reagent (for 11 2-bromothiophene 3,4-dimethoxy-phenylacetic acid 3-chlorobenzaldehyde 12 1-bromo-4- 3,4-dimethoxy-phenylacetic acid 3-chlorobenzaldehyde 13 me 2-br m o ethl-hiene -13,4-dimethoxy-phenylacetic acid '3-chlorobenzaldehyde :;14 2-bromo-5-methyl-thiophene 13,4-dimethoxy-phenylacetic acid 14-bromo-2- LibC/475183specd 1-bromo-4- 3,4-dimethoxy-phenylacetic acid 4-bromo-2- S(dimethylamino)benzene thiophencarboxaldehyde 16 5-bromo-2-methoxypyridine 3,4-dimethoxy-phenylacetic acid 3-chlorobenzaldehyde 17 5-bromo-2-methoxypyridine 3,4-dimethoxy-phenylacetic acid 3-chlorobenzaldehyde 18 5-bromo-2- 3,4-dimethoxy-phenylacetic acid 3-chlorobenzaldehyde ___dimethylaminopyridine Table Examples 11-18 Ex. No. Name Analytical Data 11 4-amino-5-(3-chlorophenyl)-6-(3,4-dimethoxyphenyl)-7-(thiophen- IR: 3440, 2940, 1540, 1420, 1020 2-yI)pyddo [2,3-dipyrimidine hydrochloride cm- 1 MS m/z 475 12 4-amino-5-(3-chlorophenyl)-6-(3,4-dimethoxyphenyl)-7-(4- IR: 3440, 2940, 1600, 1570, 1360, (dimethylamino)phenyl)pyrido [2,3-dipyrimidine dihydrochloride 1170; MS m/z 512 13 4-amino-5-(3-chlorophenyl)-6-(3,4-dimethoxyphenyl)-7-(5- IR: 3440, 2920, 1600, 1570, 1440, methylthiophen-2-yl)pyrido [2,3-dipyrimidine hydrochloride 1360; MS m/z 489 (M+H)Y.

14 4-amino-5-(4-bromothiophen-2-yl)-6-(3,4-dimethoxyphenyl)-7-(5- IR: 3420, 2960, 1580, 1440, 820; methylthiophen-2-yl)pyrido [2,3-dipyrimidine hydrochloride MS m/z 539/541 4-amino-5-(4-bromothiophen-2-yl)-6-(3,4-dimethoxyphenyl)-7-(4- IR: 3420, 2960, 1580, 1440, 820; (dimethylamino)phenyl)pyrido [2,3-dipyrimidine dihydrochioride MS m/z 539/541 16 4-amino-5-phenyl-6-(3,4-dimethoxyphenyl)-7-(5-methoxy-2- IR: 3420, 2960, 1580, 1440, 820; pyridinyl)pyrido [2,3-dipyrimidine hydrochloride MIS m/z 539/541 17 4-amino-5-(3-chlorophenyl)-6-(3,4-dimethoxyphenyl)-7-(5- IR: 3420, 2960, 1580, 1440, 820; methoxy-2-pyridinyl)pyrido [2,3-d]pyrimidine hydrochloride MS m/z 539/541 18 4-amino-5-(3-chlorophenyl)-6-(3,4-dimethoxyphenyl)-7-(5- IR: 3420, 2960, 1580, 1440, 820; dimethylamino-2-pyridinyl)pyrido [2,3-dipyrimidine hydrochloride ,MS m/z 539/541 Example 19 4-amino-5,6-(bis-4-(2-propyl)phenyl)-7-(4-dimethylaminophenyl)pyrido[2,3-d]pyrimidine 5 A sample of 4,6-diamino-5-(1,2-bis(4-(2-propyl)phenyl)ethenyl)pyrimidine (745mg, 2mmol) was dissolved in 2OmL- 1 ,2,4-trichlorobenzene containing 4-dimethylaminobenzaldehyde (0.89g, 6mmol), and approximately 1g of 4A molecular sieves were added to the reaction mixture. The mixture was heated to reflux for 20h, cooled, and filtered through a pad of celite. The filtrate was applied directly to a silica gel chromatography column, which was eluted with 2.5% (19:1 ethanol:ammoniumn hydroxide) in ethyl acetate to give the desired product (186mg, 18.5% yield): IR 3460, 2960, 1605, 1555, 1540, 1525, 1350, 820; MS mlz 502 The 4,6-d iamino-5-( 1,2-bis(4-isopropylphenyl)ethenyl)pyri mid ine was prepared as follows: I ,2-Bis(4-(2-propyl)phenyl)acetylene To a solution of 4-iodoisopropylbenzene (12.3g, 50mmol, Lancaster Chemical Co.) in triethylamine (15Oml-) was added trimethylsilylacetylene (5.89g, 6Ommol), dichlorobis (triphenylphosphine)palladium(ll) (0.70g, immol, Aldrich), and copper(l) iodide The reaction was stirred at room temperature for 1 8h, diluted with hexanes and filtered. The filtrate was evaporated under reduced pressure to give crude 1-(4-(2-propyl)phenyl)-2-trimethylsily acetylene.

The crude 1 -(4-(2-propyl)phenyl)-2-trimethylsilylacetylene was dissolved in methanol (1 0ml-).

Aqueous 1 M potassium carbonate solution (25mL) was added and the reaction stirred at room temperature for 2h. The reaction mixture was then diluted with water and extracted with pentane.

The organic layers were combined, dried with magnesium sulfate, and evaporated under reduced ST pressure without heating to give crude 4-(2-propyl)phenylacetylene.

LibC/4751 83speci '1 The crude 4-isopropyiphenylacetylene was dissolved in triethylamine (1 0mL-). 4iodoisopropylbenzene (1 2.3g, 50mmol), d ich lorobis(tri phenyl phosph ine)pal lad ium(l11) (0.70g, I mmol), and copper(I) iodide (1.5g) were added. The reaction was stirred at room temperature for 2 days, heated to reflux for 1h, cooled, diluted with hexanes, and filtered. The filtrate was evaporated under reduced pressure. The residue was filtered through a pad of silica gel with hexanes, and the solvent was evaporated to give 11.40 g of 1 ,2-bis(4-(2-propyl)phenyl)acetylene.

4.6-diamino-5-(1 ,2-bis(4-isopropylphenyl)ethenyl)pyrimidine 1 ,2-Bis(4-(2-propyl)phenyl)acetylene (11 .40g, 43mmol) was dissolved in 5OmL- THE, catecholborane (1M, 50mL-) in THE was added, and the mixture was heated at reflux for 30h. The mixture was cooled, then 4,6-diamino-5-iodo-pyrimidine, 3OmL- saturated aqueous sodium bicarbonate, 20mL- 3N aqueous sodium hydroxide, and 1 .00g (0.87mmol) tetrakis(triphenylphosphine) palladium(0) were added. The mixture was heated to reflux for 18h, cooled, diluted with water, then extracted with ethyl acetate. The organic layers were combined, dried with magnesium sulfate, and the solvent evaporated. The residue was chromatographed on silica gel with 2.5% to 5% (19:1 15 ethanol:ammonium hydroxide) in ethyl acetate to give the desired product (4.53 g, 28% yield).

Examples 20-25 Following the procedures of Example 19, except substituting the appropriate reagents required for R 3

R

2 and R 1 as indicated in Table 6 below, compounds of Examples 26-3 2 were prepared as described in Table 7 below.

20 Table 6 Examples 20-25 Ex. No. R 3 Reagent (for 7-position) R 2

R

1 Reagent (for 5 and 6-positions) 4-dimethylamino-benzaldehyde 1 ,2-diphenylacetylene 21 4-dimethylamino-benzaidehyde 1 ,2-bis(3-fluorophenyl)acetylene 22 4-dimethylamino-benzaldehyde 1 ,2-bis(3,4-dimethoxyphenyl)acetylene 23 4-dimethylamino-benzaldehyde 1 ,2-bis(3-fluoro-4-methylphenyl)acetylene 24 thiophen-2-carboxyaldehyde 1 ,2-bis(3-fluoro-4-methylphenyl)acetylene 25 Ithiophen-2-carboxaldehyde 1 ,2-diphenylacetylene Table 7 Examples 20-25 Ex. No.IName Analytical Data 4-amino-5,6-diphenyl-7-(4-dimethylaminophenyl)pyrido IR: 3410, 1635, 1600, 1580, 1360, -dipyrimidine dihydrochloride 705 cm- 1 MS m/z 418 21 4-amino-5,6-bis(3-fluorophenyl)-7-(4-dimethylaminophenyl-pyido IR: 3450, 3060, 1605, 1540, 1345, [2,3-dlpyrimidine 1200 cm- 1 MS m/z 454 22 4-amino-5,6-bis(3,4-dimethoxyphenyl)-7-(4- IR: 3400, 3100-2800, 1630, 1600, dimethylaminophenyl)pyrido [2,3-d]pyrimidine trihydrochlonide 1575, 1510, 1360, 1250, 1140, MS m/z 538 23 4-amino-5,6-bis(3-fluoro-4-methylphenyl)-7-(4- IR: 3330, 3100-2800, 1635, 1600, dimethylaminophenyl)pyddo [2,3-d]pyrimidine dihydrochloride 1575, 1535, 1505, 1360, 1200 cm- 1 m/z 482 24 4-amino-5,6-bis(3-fluoro-4-methylphenyl)-7-(thiophen-2-yl)pyrido IR: 3330, 3100-2800, 1635, 1580, [2,3-dpyrimidine hydrochloride 1540, 1505, 1415, 1365, 1235 cm- 1 m/z 445 4-amino-5,6-diphenyl-7-(thiophen-2-yl)pyndo [2,3-dipyrimidine IR: 3470, 3390, 3050, 1540, 1420 cm-'

I*.

z LibC/475lB3speci 23 Examples 26-30 Following the procedures of Examples 10, except substituting the appropriate reagents required for R 3

R

2 and RI as indicated in Table 8 below, compounds of Examples 26-30 were prepared as described in Table 9 below.

Table 8 Examples 26-30 Ex. No. IR 3 Reagent (for 7-position) R 2 Reagent (for 6-position) R 1 Reagent (for 26 5-bromo-2-(dimethylamino)pyridine phenylacetic acid benzaldehyde 27 5-bromo-2-(dimethylamino)pyridine 3,4-dimethoxy-phenylacetic acid benzaldehyde 28 5-bromo-2-(dimethylamino)pyndine phenylacetic acid 3-chlorobenzaldehyde 29 5-bromo-2-(dimethylamino)pyridine phenylacetic acid 4-bromothiophen-2- 5-bromo-2-(dimethylamino)pyridine phenylacetic acid 3-bromobenzaldehyde Table 9 Examples 26.30 Ex. No.jName Analytical Data 26 14-amino-5,6-diphenyl-7-(5-dimethylamino-2-pyidinyl)pyrido IR: 3400, 3040, 1640, 1565, 1365 dipyrimidine hydrochloride cm- 1 MS m/z 419 27 4-amino-5-phenyl-6-(3,4-dimethoxyphenyl)-7-(5- IR: 3420, 2930, 1645, 1600, 1255; (dimethylamino)pyridin-2-yl)pyrido [2,3-dipyrimidine hydrochloride MS m/z 479 28 4-amino-5-(3-chlorophenyl)-6-phenyl-7-(5-dimethylamino-2 IR: 3420, 3040, 1650, 1575, 1260; ____pyridinyl)pyrido [2,3-djpyrimidine dihydrochloride IMS m/z 453 (M+H)Y.

29 4-amino-5-(4-bromothiophen-2-yl)-6-phenyl-7-(5-dimethylamino- IR: 3400, 3100, 1650, 1355; MS 2-pyridinyl)pyrido [2,3-d]pyrimidine hydrochloride m/z 503/505 4-amino-5-(3-bromophenyl)-6-phenyl-7-(5-dimethylamino-2- IR: 3450, 3050, 1650, 1575; MS ____yridinyl)pyrido [2,3-dpyimidine hydrochloride m/z 497/499 (M+H)Y.

Example 31 Following the procedures of Example 10, except substituting the reagents shown below for the RI and R 2 reagents, the compound shown in Table 10 below was prepared.

Table ___Example 311 Ex. No. 15-position 16-position 7-position moiet 31 14-bromo-2-thiophenecarboxaldehyde ethyl 2-cyclopropylacetate dimethylamine/6-dimethylamino- I 3-pyridine ___nalytical Data, Example 31 Ex. No. Name Analytical Data 31 1 4 -amino-5-(4-bromothienyl)-6-cyclopropyl-7-(6-dimethylamino- MS mlz 467; IR (cm- 1 3426, ___3-pyridinyl)pyrido [2,3-dpyimidine hydrochloride 3001, 1649, 1600, 1373 Examples 32-33 Following the procedures of Example 10, except substituting the reagents shown below for the

R

1 and R 2 reagents, the compounds shown in table 11 below were prepared.

Table 11 Ex amp 32-33 Ex.No 5-position {6-position ~7-position moiety 32 3-bromobenzaldehyde cyclohexylmethyl magnesium Idimethylamine/6-dimethylamino-3p~yridine 33 3-bromobenzaldehyde n-hexyl magnesium chloride dimethylamine/6-dimethylamino-3jpyridine IT Analytical Data, Examples 32-33 LibC/4751 83speci Ex. No. Name Analytical Data 32 4-amino-5-(3-bromophenyl)-6-cyclohexyl-7-(6- MS mlz 503, 505 (1 Br); IR (cmdimethylamino-3-pyridinyl)pyddo [2,3-dlpyrimidine 1) 3432, 3047, 2945, 1560, 1465, 1340 33 4-amino-5-(3-bromophenyl)-6-phenyl-7-(6-dimethylamino-3- MS mlz 491, 493 (1 Br); IR (cmpyridinyl)pyrido [2,3-dlpyrimidine hydrochloride 1)3437,3025,2952,1550,1460,1320 9* LibC/475l B3speci

Claims (14)

1. A compound or a pharmaceutically accepted salt thereof having the formula (1) NH 2 R 1 N N R(l wherein R 1 is phenyl or thienyl, each of which is unsubstituted or substituted with one or two substituents independently selected from alkyl, alkoxy or halo; R 2 is lower alkyl, cycloalkyl, phenyl or phenyl alkyl wherein each of said phenyl is unsubstituted or substituted with one or two substituents independently selected from alkyl, alkoxy or halo; and R 3 is phenyl, pyridyl or thienyl, each of which is unsubstituted or substituted with one or two substituents selected from alkoxy or di(loweralkyl)amino.

2. A compound according to claim 1 wherein R 3 is selected from the group consisting of: 4- (dimethylamino)phenyl; 5-dimethylamino-2-pyridinyl; 5-methoxy-2-pyridinyl; 4-methoxyphenyl; methylthiophen-2-yl; and thiophen-2-yl.

3. A compound according to claim 1 or claim 2 wherein R 2 is selected from the group consisting of: ethyl; 3-fluorophenyl; 3-fluoro-4-methylphenyl; 3,4-dimethoxyphenyl; 3-methoxyphenyl;

4-methoxyphenyl; pentyl; phenyl; and 4-(2-propyl)phenyl. 15 4. A compound according to any one of claims 1 to 3 wherein RI is selected from the group consisting of: 3-bromophenyl; 3-bromo-4-fluorophenyl; 4-bromothiophen-2-yl; 3-chlorophenyl; 3,4- dimethoxyphenyl; 3-fluorophenyl; 3-fluoro-4-methylphenyl; and 4-(2-propyl)phenyl. A compound which is 4-a i *5(-rm--loohnl--etl7(-dimty mn)prd[,-~yimdi 4 -amino-5-(3-bromo-4-fluorophenyl)-6-pentyl-7-(4-impehylmn)pyrido[2,3d]pyrimidine; 4-amino-5-(3-bromo-4frphenyl)-6-ephenyl)-7-(oph -2yphyrio[,pyrimi[23dyin md e 4 -am ino-5-(3-bromophenyl)-6-(eth7-t ophen-yl) py(rmetoxyphenlpyri do[,3priidn 4-amino-5-(3-bromophenyl)-6-etyl-7-(thiophen-2-yl)pyrido[2,3-dpyrimidine; :*4-amino-5-(3-bromophenyl)-6-pen-mtyl penl-7-(thiophen-2-yl)prd [2,3-dlpyrimidine; 25 4 -amino-5-(3-bromophenyl)-6-(4-(2-propyl)phenyl)-7-(4-(dimethylamino)phenyl)pyrido[2,3- d]pyrimidine; 4 -amino-5-(3-bromophenyl)-6-(3-methoxyphenylmethyl)-7(thiophen-2yl)pyrido[2,3-d]pyrimidine; 4 -amino-5-(3-bromophenyl)-6-(3,4-dimethoxyphenyl)-7(4(dimethylamino)phenyl)pyrido[2,3. d]pyrimidine; 4-amino-5-(4-bromthiophen-2-yl)-6-(3,4-d imethoxyphenyl)-7-(thiophen-2-yl)pyrido 3-d]pyrimidime; 4 -amino-5-(3-chlorophenyl)-6-(3,4-dimethoxyphenyl)-7(th iophen-2-yl)pyrido [2,3-d]pyrimidime; 4 -amino-5-(3-chlorophenyl)-6-(3,4-dimethoxyphenyl)7(4(dimethylamino)phenyl)pyrido [2,3- d]pyrimidine; 4 -amino-5-(3-chlorophenyl)-6-(3,4-dimethoxyphenyl)7(5-methylthiophen-2yl)pyrido [2,3- dlpyrimidine; 4 -amino-5-(4-bromothiophen-2-yl)-6-(3,4-dimethoxyphenyly7(5methylthiophen-2yl)pyrido [2,3- LibC/4751 83speci 4-amino-5-(4-bromothiophen-2-yI)-6-(3,4-dimethoxyphenyl)-7-(4-(dimethylamino)phenyl)pyrido [2,3- dipyrimidine; 4-amino-5-phenyl-6-(3,4-dimethoxyphenyl)-7-(5-methoxy-2-pyridinyl)pyrido [2,3-d]pyrimidine; 4-amino-5-(3-chlorophenyl)-6-(3,4-dimethoxyphenyl)-7-(5-methoxy-2-pyridinyl)pyrido [2,3- djpyrimidine; 4-amino-5-(3-chiorophenyl)-6-(3,4-dimethoxyphenyl)-5-(5-d imethylamino-2-pyridinyl)pyrido [2,3- dipyrimidine; 4-amino-5,6-bis(4-(2-propyl)phenyl-7-(4-dimethylaminopheny)pyrido [2,3-dipyrimidine; 4-amino-5,6-diphenyl-7-(4-dimethylaminophenyl)pyrido [2,3-dlpyrimidine; 4-amino-5,6-bis(3-fluorophenyl)-7-(4-dimethylaminophenyl)pyrido [2,3-d]pyrimidine; 4-amino-5,6-bis(3,4-dimethoxyphenyl)-7-(4-dimethylaminophenyl)pyrido [2,3-dlpyrimidine; 4-amino-5,6-bis(3-fluoro-4-methylphenyl)-7-(4-dimethylaminophenyl)pyrido [2,3-dipyrimidine; 4-amino-5,6-bis(3-fluoro-4-methylphenyl)-7-(thiophen-2-yl)pyrido [2,3-dipyrimidine; 4-amino-5,6-diphenyl-7-(thiophen-2-yl)pyrido [2,3-djpyrimidine; 15 4-amino-5,6-diphenyl-7-(5-dimethylamino-2-pyridinyl)pyrido [2,3-d]pyrimidine; 4-mn hnl6(,-iehxpey)*-5(iehlmn~ yii--lprd 0O3dpriiie 4-amino-5-(3crphenyl -etophenyl-7-(5-(dimethylaminopyridin2yl)pyrido [2,3-dipyrimidine; 4-amino-5-(3-crohloophen2yl)-6-phenyl-7-(5-dimethylamino-2-pyrid inyl)pyrido [2,3-d]pyrimidine; 4-amino-5-(4-bromtophen2yI)-6-phenyl-7-(5-dimethylamino-2-pyridinyl)pyrido [2,3-dipyrimidme; 4-am ino-5-(3-bromophenyl)-6-pyheyl-7-(6-d imethylam ino-3-pyrid inyl)pyrido [2,3-dpyri midine; o 4-amino-5-(3-bromophenyl)-6-cyleyl-7-(6-dimethylamino-3-pyridinyl)pyrido [2,3d]pyrimidine; o

6. A 5,6,7-trisubstituted-4-aminopyridol[2,3-d]pyrimidine derivative, substantially as hereinbefore described with reference to any one of the examples.

7. A pharmaceutical composition comprising a therapeutically effective amount of a 25 compound according to any one of claims 1 to 6 in combination with a pharmaceutically acceptable carrier. A method for inhibiting adenosine kinase by administering a compound according to any one of claims 1 to 6 or a composition according to claim 7.

9. A compound according to any one of claims 1 to 6 or a composition according to claim 7 when used in inhibiting adenosine kinase. The use of a compound according to any one of claims 1 to 6 for the manufacture of a medicament for inhibiting adenosine kinase.

11. A method of treating ischaemia, neurological disorders, nociperception, inflammation, immunosuppression, gastrointestinal dysfunctions, diabetes and sepsis in a mammal in need of such treatment, comprising administering to the mammal a therapeutically effective amount of a compound according to any one of claims 1 to 6 or a composition according to claim 7.

12. A compound according to any one of claims 1 to 6 or a composition according to claim 7 when used in treating ischaemia, neurological disorders, nociperception, inflammation, immunosuppression, gastrointestinal dysfunctions, diabetes and sepsis. LibC/4761 83speci

13. The use of a compound according to any one of claims 1 to 6 for the manufacture of a medicament for treating ischaemia, neurological disorders, nociperception, inflammation, immunosuppression, gastrointestinal dysfunctions, diabetes and sepsis.

14. A method, compound, composition or use according to any one of claims 11 to 13, wherein cerebral ischaemia, myocardial ischaemia, angina, coronary artery bypass graft surgery, percutaneous transluminal angioplasty, stroke, thrombotic and embolic conditions, epilepsy, anxiety, schizophrenia, pain perception, neuropathic pain, visceral pain, arthritis, sepsis, diabetes and abnormal gastrointestinal motility is treated. A process for the preparation of a compound having the formula NH 2 R' N 'R 2 N N R 3 wherein R 1 is phenyl or thienyl, each of which is unsubstituted or substituted with one or two S: substituents independently selected from the group consisting of alkyl, alkoxy, and halo; R 2 is loweralkyl, cycloalkyl, phenyl or phenylalkyl, wherein each ,of the foregoing phenyl groups is unsubstituted or substituted with one or two substituents independently selected from the group is consisting of alkyl, alkoxy, and halo; and R 3 is phenyl, pyridyl, or thienyl, each of which is unsubstituted or substituted with one or two substituents independently selected from the group consisting of alkoxy and di(loweralkyl)amino; the process comprising; reacting an aryl, heteroaryl, or a heterocyclic bromide having the formula R 3 -Br wherein R 3 is as *i *defined above with a carboxylic acid derivative having the formula R 2 -CH 2 -CO-Y, wherein Y is OH or 20 CI, and R 2 is as defined above, with N,O-dimethylhydroxylamine hydrochloride, 1-(3- dimethylaminopropyl)-3-ethylcarbodiimide or 1-hydroxybenzotriazole hydrate and triethylamine, and isolating a first intermediate compound having the formula R 3 -CO-CH 2 -R2; reacting the first intermediate compound having the formula R 3 -CO-CH 2 -R 2 with an aldehyde having the formula R 1 -CHO, wherein R 1 is as defined above, and malononitrile in the presence of an ammonium salt under anhydrous conditions, and isolating a second intermediate compound having the formula; R 1 NC R 2 H 2 N N R 3 reacting the second intermediate compound with formamide at reflux for from about 1 to about 24h, and isolating the compound of formula I.

16. A process for the preparation of compounds having the formula NH 2 R 1 N I R 2 S^ NR 3 No N. R\ LibC/475183speci wherein R 1 is phenyl or thienyl, each of which is unsubstituted or substituted with one or two substituents independently selected from the group consisting of alkyl, alkoxy, and halo; R 2 is loweralkyl, cycloalkyl, phenyl or phenylalkyl, wherein each of the foregoing phenyl groups is unsubstituted or substituted with one or two substituents independently selected from the group consisting of alkyl, alkoxy, and halo; and R 3 is phenyl, pyridyl, or thienyl, each of which is unsubstituted or substituted with one or two substituents independently selected from the group consisting of alkoxy and di(loweralkyl)amino; the process comprising reacting an aryl, heteroaryl, or a heterocyclic bromide having the formula R 3 -Br with a carboxylic acid derivative having the formula R 2 -CH 2 -CO-Y, wherein Y is OH or CI, and R 2 is defined above, with N,O-dimethylhydroxylamine hydrochloride 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide or 1- hydroxybenzotriazole hydrate and triethylamine, and isolating a first intermediate compound having the formula R 3 -CO-CH 2 -R2; treating the first intermediate compound having the formula R 3 -CO-CH 2 -R 2 with a compound R 1 having the formula NC CN, wherein R 1 is as described above, at reflux in an alcoholic solvent, and NC R 2 15 isolating a second intermediate product having the formula H 2 N R and reacting the second intermediate compound with sulfuric acid and heating followed by treatment with methyl orthoformate at reflux for from about 1 to about 24h, and isolating a third intermediate compound having the structure OH R 1 2 *1R Q 3 N N R3 20 treating the third intermediate compound with a chlorinating agent, and isolating a fourth intermediate product having the formula CI R 1 R 2 N N R 3 treating the fourth intermediate compound with ammonia, and isolating the compound of formula I.

18. A compound of the formula NH 2 R 1 R 2 N N R 3 wST herein R 1 R 2 and R 3 are as defined in claim 1 and X is OH or halogen. LibC/475183speci

19. A method of using a compound of formula II according to claim 18, as an intermediate to produce a compound of formula I according to claim 1, said method comprising, reacting said compound of formula II with ammonium, under the appropriate conditions, to form a compound of formula I according to claim 1. Dated 26 September 2001 ABBOTT LABORATORIES Patent Attorneys for the Applicant/Nominated Person SPRUSON FERGUSON S 9* r 9* 9 66 S* 0:0$0 00 0 **r ft LibC/475183speci