the compounds described above can be prepared by methods well known in the art , as well as by the synthetic routes disclosed herein . for example , a pyrimidine compound can be prepared by using 2 , 4 , 6 - trichloro - pyrimidine as a starting material . the three chloro groups can be displaced by various substitutes . more specifically , first chloro group ( e . g ., at position 6 ) can react with , e . g ., morpholine , to form a morpholinyl pyrimidine . 2 - aryl and 2 - alkylpyrimidinde dichloro compounds can also be prepared by reacting an amidine with a malonic ester followed by treatment with phosphorous oxychloride . second chloro group can be replaced by reacting with a nucleophile , such as an alcohol in the presence of base , e . g ., sodium hydride . isomeric forms may be produced . the desired isomeric product can be separated from others by , e . g ., high performance liquid chromatography . third chloro group undergoes a displacement reaction with , e . g ., hydrazine , and the primary amine of the coupled hydrazine moiety further reacts with an aldehyde . thus , a pyrimidine compound of this invention is obtained . the chemicals used in the above - described synthetic routes may include , for example , solvents , reagents , catalysts , and protecting group and deprotecting group reagents . the methods described above may also additionally include steps , either before or after the steps described specifically herein , to add or remove suitable protecting groups in order to ultimately allow synthesis of the pyrimidine compounds . in addition , various synthetic steps may be performed in an alternate sequence or order to give the desired compounds . synthetic chemistry transformations and protecting group methodologies ( protection and deprotection ) useful in synthesizing applicable pyrimidine compounds are known in the art and include , for example , those described in r . larock , comprehensive organic transformations , vch publishers ( 1989 ); t . w . greene and p . g . m . wuts , protective groups in organic synthesis , 2 nd ed ., john wiley and sons ( 1991 ); l . fieser and m . fieser , fieser and fieser &# 39 ; s reagents for organic synthesis , john wiley and sons ( 1994 ); and l . paquette , ed ., encyclopedia of reagents for organic synthesis , john wiley and sons ( 1995 ) and subsequent editions thereof . a pyrimidine compound thus obtained can be further purified by flash column chromatography , high performance liquid chromatography , or crystallization . also within the scope of this invention is a pharmaceutical composition that contains an effective amount of one or more of the pyrimidine compounds of this invention and a pharmaceutically acceptable carrier . further , the present invention covers a method of administering an effective amount of such a compound to a subject in need of treatment of il - 12 overproduction related diseases ( e . g ., rheumatoid arthritis , sepsis , crohn &# 39 ; s disease , multiple sclerosis , psoriasis , or insulin - dependent diabetes mellitus ). “ an effective amount ” refers to the amount of the compound which is required to confer a therapeutic effect on the treated subject . the interrelationship of dosages for animals and humans ( based on milligrams per meter squared of body surface ) is described in freireich et al ., ( 1966 ) cancer chemother rep 50 : 219 . body surface area may be approximately determined from height and weight of the patient . see , e . g ., scientific tables , geigy pharmaceuticals , ardley , n . y ., 1970 , 537 . an effective amount of the pyrimidine compound of this invention can range from about 0 . 001 mg / kg to about 1000 mg / kg . effective doses will also vary , as recognized by those skilled in the art , depending on the diseases treated , route of administration , excipient usage , and the possibility of co - usage with other therapeutic treatments such as use of other agents . to practice the method of the present invention , a pyrimidine compound , as a component of a pharmaceutical composition , can be administered orally , parenterally , by inhalation spray , topically , rectally , nasally , buccally , vaginally or via an implanted reservoir . the term “ parenteral ” as used herein includes subcutaneous , intracutaneous , intravenous , intramuscular , intraarticular , intraarterial , intrasynovial , intrastemal , intrathecal , intralesional and intracranial injection or infusion techniques . a sterile injectable composition , for example , a sterile injectable aqueous or oleaginous suspension , can be formulated according to techniques known in the art using suitable dispersing or wetting agents ( such as , for example , tween 80 ) and suspending agents . the sterile injectable preparation can also be a sterile injectable solution or suspension in a non - toxic parenterally acceptable diluent or solvent , for example , as a solution in 1 , 3 - butanediol . among the acceptable vehicles and solvents that can be employed are mannitol , water , ringer &# 39 ; s solution and isotonic sodium chloride solution . in addition , sterile , fixed oils are conventionally employed as a solvent or suspending medium ( e . g ., synthetic mono - or diglycerides ). fatty acids , such as oleic acid and its glyceride derivatives are useful in the preparation of injectables , as are natural pharmaceutically - acceptable oils , such as olive oil or castor oil , especially in their polyoxyethylated versions . these oil solutions or suspensions can also contain a long - chain alcohol diluent or dispersant , or carboxymethyl cellulose or similar dispersing agents . other commonly used surfactants such as tweens or spans or other similar emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid , liquid , or other dosage forms can also be used for the purposes of formulation . a composition for oral administration can be any orally acceptable dosage form including , but not limited to , capsules , tablets , emulsions and aqueous suspensions , dispersions and solutions . in the case of tablets for oral use , carriers which are commonly used include lactose and corn starch . lubricating agents , such as magnesium stearate , are also typically added . for oral administration in a capsule form , useful diluents include lactose and dried corn starch . when aqueous suspensions or emulsions are administered orally , the active ingredient can be suspended or dissolved in an oily phase combined with emulsifying or suspending agents . if desired , certain sweetening , flavoring , or coloring agents can be added . a nasal aerosol or inhalation composition can be prepared according to techniques well - known in the art of pharmaceutical formulation and can be prepared as solutions in saline , employing benzyl alcohol or other suitable preservatives , absorption promoters to enhance bioavailability , fluorocarbons , and / or other solubilizing or dispersing agents known in the art . a pyrimidine compound of this invention can also be administered in the form of suppositories for rectal administration . the carrier in the pharmaceutical composition must be “ acceptable ” in the sense of being compatible with the active ingredient of the formulation ( and preferably , capable of stabilizing it ) and not deleterious to the subject to be treated . for example , solubilizing agents such as cyclodextrins , which form specific , more soluble complexes with the compounds of this invention , or one or more solubilizing agents , can be utilized as pharmaceutical excipients for delivery of the pyrimidine compounds . examples of other carriers include colloidal silicon dioxide , magnesium stearate , cellulose , sodium lauryl sulfate , and d & amp ; c yellow # 10 . the biological activities of a pyrimidine compound can be evaluated by a number of cell - based assays . one of such assays can be conducted using cells from human peripheral blood mononuclear cells ( pbmc ) or human monocytic cell line ( thp - 1 ). the cells are stimulated with a combination of human interferon - γ ( ifnγ ) and lipopolysaccharide or a combination of ifnγ and staphylococcus aureus cowan i in the presence of a test compound . the level of inhibition of il - 12 production can be measured by determining the amount of p70by using a sandwich elisa assay with anti - human il - 12 antibodies . ic 50 of the test compound can then be determined . specifically , pbmc or thp - 1 cells are incubated with the test compound . cell viability was assessed using the bioreduction of mts [ 3 -( 4 , 5 - dimethylthiazol - 2 - yl )- 5 -( 3 - carboxymethoxyphenyl )- 2 -( 4 - sulfophenyl )- 2h - tetrazolium ] ( promega , madison , wis .). a pyrimidine compound can also be evaluated by animal studies . for example , one of such studies involves the ability of a test compound to treat adjuvant arthritis ( i . e ., a il - 12 overproduction related disorder ) in rats . without further elaboration , it is believed that the above description has adequately enabled the present invention . the following specific embodiments are , therefore , to be construed as merely illustrative , and not limitative of the remainder of the disclosure in any way whatsoever . all of the publications cited herein are hereby incorporated by reference in their entirety . to a solution of 4 -[ 6 - chloro - 2 -( 2 - pyridin - 2 - yl - ethoxy )- pyrimidin - 4 - yl ]- morpholine ( 1 . 61 g , 5 . 0 mmol ) in ch 2 cl 2 ( 40 ml ) was added methanol ( 10 ml ) followed by the addition of mcpba ( 70 %, 1 . 43 g , 5 . 8 mmol ) in one portion . the reaction mixture was stirred overnight at room temperature , affording a clear solution . the solution was cast into saturated aqueous nahco 3 ( 35 ml ) then the organic phase was separated , washed with 10 % aqueous na 2 s 2 o 3 ( 40 ml ) and brine ( 40 ml ), and dried ( na 2 so 4 ), filtered and evaporated in vacuo to give a pure product , 4 -{ 6 - chloro - 2 -[ 2 -( 1 - oxy - pyridin - 2 - yl )- ethoxy ]- pyrimidin - 4 - yl }- morpholine as a white solid , ( 1 . 46 g , 86 . 7 %). 1 h - nmr ( cdcl 3 ) ( ppm ), j ( hz ): 8 . 25 - 8 . 23 ( m , 1h ), 7 . 41 - 7 . 7 . 38 ( m , 1h ), 7 . 20 - 7 . 16 ( m , 2h ), 6 . 14 ( s , 1h ), 4 . 71 ( t , j = 6 . 0 , 2h ), 3 . 77 - 3 . 73 ( m , 4h ), 3 . 63 - 3 . 55 ( m , 4h ), 3 . 40 ( t , j = 6 . 0 , 2h ), anhydrous hydrazine ( 0 . 640 ml , 20 mmol ) was added to a solution of 4 -{ 6 - chloro - 2 -[ 2 -( 1 - oxy - pyridin - 2 - yl )- ethoxy ]- pyrimidin - 4 - yl })- morpholine ( 1 . 35 g , 4 . 0 mmol ) in dioxane ( 15 ml ) under the nitrogen protection . the obtained mixture was heated at 95 - 100 ° c . for 2 h . after it was cooled down , the solvent was evaporated in vacuo until the white solid began to precipitate ( to a half the original volume ), and then h 2 o ( 15 ml ) was added . the resulting precipitate was collected by filtration and washed with water ( until the ph was neutral ). { 6 - morpholin - 4 - yl - 2 -[ 2 -( 1 - oxy - pyridin - 2 - yl )- ethoxy ]- pyrimidin - 4 - yl }- hydrazine ( 1 . 02 g ) has been obtained in 76 . 7 % yield . 1 h - nmr ( dmso - d 6 ) ( ppm ), j ( hz ): 8 . 25 ( bs , 1h ), 7 . 66 ( s , 1h ), 7 . 44 - 7 . 41 ( m , 1h ), 7 . 33 - 7 . 25 ( m , 2h ), 5 . 59 ( s , 1h ), 4 . 46 ( t , j = 6 . 0 , 2h ), 3 . 64 - 3 . 61 ( m , 4h ), 3 . 41 - 3 . 38 ( m , 4 ), 3 . 17 ( t , j = 6 ., 2h ), to a solution { 6 - morpholin - 4 - yl - 2 -[ 2 -( 1 - oxy - pyridin - 2 - yl )- ethoxy ]- pyrimidin - 4 - yl }- hydrazine ( 820 mg , 2 . 46 mmol ) and m - tolualdehyde ( 97 %, 320 mg , 2 . 58 mmol ) in methanol ( 7 ml ) acetic acid ( 2 drops ) was added . the reaction mixture was heated under reflux for 15 min . upon cooling to room temperature , a precipitating has been formed , and the solid was collected by filtration , washed with little amount of methanol and et 2 o , and dried to afford 950 mg ( 89 %) of n -( 3 - methyl - benzylidene )- n ′-{ 6 - morpholin - 4 - yl - 2 -[ 2 -( 1 - oxy - pyridin - 2yl )- ethoxy ]- pyrimidin - 4 - yl }- hydrazine as a white solid ( m . p . 187 - 188 ° c .). 1 h nmr ( 300 mhz , cdcl 3 ), δ ( ppm ): 10 . 86 ( s , 1h ), 8 . 28 - 8 . 26 ( m , 1h ), 7 . 98 ( s , 1h ), 7 . 50 - 7 . 43 ( m , 3h ), 7 . 33 - 7 . 26 ( m , 3h ), 7 . 17 ( d , j = 7 . 8 hz , 1h ), 6 . 05 ( s , 1h ), 4 . 53 ( t , j = 6 . 3 hz , 2h ), 3 . 68 - 3 . 64 ( m , 4h ), 3 . 54 - 3 . 50 ( m , 4h ), 3 . 21 ( t , j = 6 . 3 , 2h ), 2 . 33 ( s , 3h ); esms calcd for c 23 h 26 n 6 o 3 : 434 . 21 ; found : 457 . 2 ( m + na ) + . reagents . staphylococcus aureus cowan i ( sac ) was obtained from calbiochem ( la jolla , calif . ), and lipopolysaccharide ( lps , serratia marscencens ) was obtained from sigma ( st . louis , mo .). human and mouse recombinant ifn - γ were purchased from boehringer mannheim ( mannheim , germany ) and pharmingen ( san diego , calif . ), respectively . human in vitro assay . human pbmc were isolated by centrifugation using ficoll - paque ( pharmacia biotech , uppsala , sweden ) and prepared in rpmi medium supplemented with 10 % fetal calf serum ( fcs ), 100 u / ml penicillin , and 100 μg / ml streptomycin . pbmc were plated in wells of a 96 - well plate at a concentration of 5 × 10 5 cells / well , and primed by adding ifnγ ( 30 u / ml ) for 22 h and stimulated by adding lps ( 1 μg / ml ), or by adding ifnγ ( 100 u / ml ) and then stimulated by adding sac ( 0 . 01 %). a test pyrimidine compound was dissolved in dmso , and added to wells of the 96 - well plate . the final dmso concentration was adjusted to 0 . 25 % in all cultures , including the compound - free control . human thp - 1 cells were plated in wells , primed by adding ifnγ ( 100 u / ml ) for 22 h and stimulated by adding sac ( 0 . 025 %) in the presence of different concentrations of the pyrimidine compound . cell - free supernatants were taken 18 h later for measurement of cytokines . cell viability was assessed using the bioreduction of mts . cell survival was estimated by determining the ratio of the absorbance in compound - treated groups versus compound - free control . the supematant was assayed for the amount of il - 12p40 , il - 12p70 , or il - 10 by using a sandwich elisa with anti - human antibodies , i . e ., a human il - 12 p40 elisa kit from r & amp ; d systems ( berkeley , calif . ), and a human il - 12 p70 or il - 10 elisa kit from endogen ( cambridge , mass .). assays were based on the manufacturer &# 39 ; s instructions . murine in vitro assay . balb / c mice ( taconic , germantown , n . y .) were immunized with mycobacterium tuberculosis h37ra ( difco , detroit , mich .). the splenocytes were harvested 5 days and prepared in rpmi medium supplemented with 10 % fcs and antibiotics in a flat bottom 96 - well plate with 1 × 10 6 cells / well . the splenocytes were then stimulated with a combination of ifnγ ( 60 ng / ml ) and sac ( 0 . 025 %) [ or lps ( 20 μg / ml )] in the presence of a test compound . cell - free supernatants were taken 24 h later for the measurement of cytokines . the preparation of compound and the assessment of cell viability were carried out as described above . mouse il - 12 p70 , il - 10 , il - 1β , and tnfα were measured using elisa kits from endogen , according to the manufacturer &# 39 ; s instructions . the biological activities of pyrimidine compounds were tested on human pbmc or thp - 1 cells . unexpectedly , compound 1 had an ic 50 value as low as 1 . 4 nm when tested on human pbmc cells . treatment of adjuvant arthritis in rats : adjuvant arthritis ( aa ) was induced in female lewis rats by the intracutaneous injection ( base of the tail ) of 0 . 1 ml of a 10 mg / ml bacterial suspension made from ground , beat - killed mycobacterium tuberculosis h37ra suspended in incomplete freund &# 39 ; s adjuvant . rats were given a test compound orally once a day for 12 days , starting the day following the induction . the development of polyarthritis was monitored daily by macroscopic inspection and assignment of an arthritis index to each animal , during the critical period ( days 10 to 25 post - immunization ). the intensity of polyarthritis was scored according to the following scheme : ( a ) grade each paw from 0 to 3 based on erythema , swelling , and deformity of the joints : 0 for no erythema or swelling ; 0 . 5 if swelling is detectable in at least one joint ; 1 for mild swelling and erythema ; 2 for swelling and erythema of both tarsus and carpus ; and 3 for ankylosis and bony deformity . maximum score for all 4 paws was thus 12 . ( b ) grade for other parts of the body : for each ear , 0 . 5 for redness and another 0 . 5 if knots are present ; 1 for connective tissue swelling ( saddle nose ); and 1 for the presence of knots or kinks in the tail . the highest possible arthritic index was 16 . experiments with the aa model were repeated four times . oral administration of pyrimidine compounds of this invention reproducibly reduced the arthritic score and delayed the development of polyarthritis in a dose - dependent manner . the arthritis score used in this model was a reflection of the inflammatory state of the structures monitored and the results therefore show the ability of the test compound to provide relief for this aspect of the pathology . treatment of crohn &# 39 ; s disease in dinitrobenzene sulfonic acid - induced inflammatory bowel syndrome model rats : wistar derived male or female rats weighing 200 ± 20 g and fasted for 24 hours were used . distal colitis was induced by intra - colonic instillation of 2 , 4 - dinitrobenzene sulfonic acid ( dnbs , 25 mg in 0 . 5 ml ethanol 30 %) after which air ( 2 ml ) was gently injected through the cannula to ensure that the solution remained in the colon . a test compound and / or vehicle was administered orally 24 and 2 hours before dnbs instillation and then daily for 5 days . one control group was similarly treated with vehicle alone while the other is treated with vehicle plus dnbs . the animals were sacrificed 24 hours after the final dose of test compound administration and each colon was removed and weighed . colon - to - body weight ratio was then calculated for each animal according to the formula : colon ( g )/ bw × 100 . the “ net ” increase in ratio of vehicle - control + dnbs group relative to vehicle - control group was used as a base for comparison with test substance treated groups and expressed as “% deduction .” a 30 % or more reduction in colon - to - body weight ratio , relative to the vehicle treated control group , was considered significant . unexpectedly , compound 1 had about 63 % reduction . rats treated with test substance orally showed a marked reduction in the inflammatory response . these experiments were repeated three times and the effects were reproducible . treatment of crohn &# 39 ; s disease in cd4 + cd45rb high t cell - reconstituted scid colitis model mice : spleen cells were prepared from normal female balb / c mice . for cell purification , the following anti - mouse antibodies were used to label non - cd4 + t cells : b220 ( ra3 - 6b2 ), cd11b ( m1 / 70 ), and cd8α ( 53 - 6 . 72 ). all antibodies were obtained from biosource ( camarnllo , calif .). m450 anti - rat igg - coated magnetic beads ( dynal , oslo , norway ) were used to bind the antibodies and negative selection was accomplished using an mpc - 1 magnetic concentrator . the enriched cd4 + cells were then labeled for cell sorting with fitc - conjugated cd45rb ( 16a , pharmingen , san diego , calif .) and pe - conjugated cd4 ( ct - cd4 , caltag , burlingame , calif .). cd4 + cd 45 high cells were operationally defined as the upper 40 % of cd45rb - staining cd4 + cells and sorted under sterile conditions by flow cytometry . harvested cells were resuspended at 4 × 10 6 / ml in pbs and injected 100 μl intraperitoneally into female c . b - 17 scid mice . pyrimidine compounds of this invention and / or vehicle was orally administered once a day , 5 days per week , starting the day following the transfer . the transplanted scid mice were weighed weekly and their clinical condition was monitored . colon tissue samples were fixed in 10 % buffered formalin and embedded in paraffin . sections ( 4 μm ) collected from ascending , transverse , and descending colon were cut and stained with hematoxylin and eosin . the severity of colitis was determined based on histological examination of the distal colon sections , whereby the extent of colonic inflammation was graded on a scale of 0 - 3 in each of four criteria : crypt elongation , cell infiltration , depletion of goblet cells , and the number of crypt abscesses . lp lymphocytes were isolated from freshly obtained colonic specimens . after removal of payer &# 39 ; s patches , the colon was washed in ca / mg - free hbss , cut into 0 . 5 cm pieces and incubated twice in hbss containing edta ( 0 . 75 mm ), dtt ( 1 mm ), and antibiotics ( amphotericin 2 . 5 μg / ml , gentamicin 50 μg / ml from sigma ) at 37 ° c . for 15 min . next , the tissue was digested further in rpmi containing 0 . 5 mg / ml collagenase d , 0 . 01 mg / ml dnase i ( boehringer manheim ), and antibiotics at 37 ° c . lp cells were then layered on a 40 - 100 % percoll gradient ( pharmacia , uppsala , sweden ), and lymphocyte - enriched populations were isolated from the cells at the 40 - 100 % interface . to measure cytokine production , 48 - well plates were coated with 10 μg / ml murine anti - cd3ε antibody ( 145 - 2c11 ) in carbonate buffer ( ph 9 . 6 ) overnight at 4 ° c . 5 × 10 5 lp cells were then cultured in 0 . 5 ml of complete medium in precoated wells in the presence of 1 μg / ml soluble anti - cd28 antibody ( 37 . 51 ). purified antibodies were obtained from pharmingen . culture supernatants were removed after 48 h and assayed for cytokine production . murine ifnγ was measured using an elisa kit from endogen ( cambridge , mass . ), according to the manufacturer &# 39 ; s instructions . histological analysis showed that oral administration of pyrimidine compounds of this invention reduced colonic inflammation as compared to vehicle control . the suppressive effect was dose - dependent with a substantial reduction at a dose of 10 mg / kg . the calculated colon - to - body weight ratio was consistent with the histological score , showing attenuation by treatment with the test compound . furthermore , analysis of cytokines from lp cells in response to anti - cd3 antibody and anti - cd28 antibody demonstrated that lp cells from vehicle control produced an augmented level of ifnγ and treatment with test substance greatly diminished the production . these results clearly demonstrated the potential of the test substance in treatment of inflammatory bowel disease represented by crohn &# 39 ; s disease . all of the features disclosed in this specification may be combined in any combination . each feature disclosed in this specification may be replaced by an alternative feature serving the same , equivalent , or similar purpose . thus , unless expressly stated otherwise , each feature disclosed is only an example of a generic series of equivalent or similar features . from the above description , one skilled in the art can easily ascertain the essential characteristics of the present invention , and without departing from the spirit and scope thereof , can make various changes and modifications of the invention to adapt it to various usages and conditions . for example , compounds structurally analogous a pyrimidine compound described in the specification also can be made , screened for their inhibiting il - 12 activities , and used to practice this invention . thus , other embodiments are also within the claims .
this invention features pyrimidine compounds of formula : r 1 is each of r 2 and r 4 is h ; r 3 is h , alkyl , aryl , heteroaryl , cyclyl , heterocyclyl , or alkylcarbonyl ; r 5 is h or alkyl ; n is 0 , 1 , 2 , 3 , 4 , 5 , or 6 ; x is nr c ; y is covalent bond , ch 2 , c , c ═ n — r c , c ═ n — or c , c ═ n — sr c , o , s , s , s , or nr c ; z is n or ch ; one of u and v is n , and the other is cr c ; and w is o , s , s , s , nr c , or ncr c ; in which each of r a and r b , independently , is h , alkyl , aryl , heteroaryl ; and r c is h , alkyl , aryl , heteroaryl , cyclyl , heterocyclyl , or alkylcarbonyl .