OA12161A - PyridoÄ2,3-dÜpyrimidine-2,7-diamine kinase inhibitors. - Google Patents

PyridoÄ2,3-dÜpyrimidine-2,7-diamine kinase inhibitors. Download PDF

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OA12161A
OA12161A OA1200200213A OA1200200213A OA12161A OA 12161 A OA12161 A OA 12161A OA 1200200213 A OA1200200213 A OA 1200200213A OA 1200200213 A OA1200200213 A OA 1200200213A OA 12161 A OA12161 A OA 12161A
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pyrido
pyrimidin
urea
piperazin
phenylamino
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Richard John Booth
Ellen Myra Dobrusin
Vara Prasad Venkata Na Josyula
Dennis Joseph Mcnamara
Peter Laurence Toogood
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Warner Lambert Co
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    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
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    • C07F9/6561—Heterocyclic compounds, e.g. containing phosphorus as a ring hetero atom containing systems of two or more relevant hetero rings condensed among themselves or condensed with a common carbocyclic ring or ring system, with or without other non-condensed hetero rings

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Abstract

Disclosed are compounds of the formula (I) wherein: R<2>, R<7>, R<13>, R<14> and R15 are independently hydrogen, or (un)substituted lower alkyl, (un)substitued lower alkenyl, (un)substituted lower alkynyl, or (un)substituted -(CH2)nR<12>; R<5> is halogen, cyano, nitro, -R<9>, -NR<9>R<10>, or -OR<9>; R<6> is halogen, cyano, nitro, -R<9>, -NR<9>R<10>, -OR<9>, -Co2R<9>, -COR<9>, -CONR<9>R<10>, -NR<9>COR<10>, (un)substituted lower alkenyl, or (un)substituted lower alkynyl; R<8> is -CO2R<13>, -COR<13>, -CONR<13>R<14>, -CSNR<13>R<14>, -C(NR<13>)NR<14>R<15>, -SO3R<13>, -S02R<13>, -SO2NR<13>R<14>, -PO3R<13>R<14>, -POR<13>R<14>, -PO(NR<13>R<14>)2; R<9> and R<10> are independently hydrogen or (un)substituted lower alkyl; R<11> is a heteroaryl or a heterocyclic group; R<12> is a cycloalkyl, a heterocyclic, an aryl, or a heteroaryl group; and n is 0,1,2, or 3. These compounds and their pharmaceutical compositions are useful for treating cell proliferative disorders, such as cancer and restenosis. These compounds are potent inhibitors of cdks and growth factor-mediated kinases.

Description

-1-
PYRIDO[2,3-djPYRIMIDINE-2.7-DIAMINE KINASE INHIBITORS
FIELD OF THE INVENTION
This invention relates to pyrido[2.3-d]pyrimidine-2.7-diamines that inhibitcvclin-dependent serine/threonine kinases and growth factor-mediated tyrosinekinase enzymes, and as such are useful to treat cell prolifération diseases anddisorders.
BACKGROUND OF THE INVENTION
Summary of the Related Art
Cell cycle kinases are naturallv occurring enzymes involveçl in régulationof the cell cycle (Meijer L.. “Chemical Inhibitors of Cyciin-Dependent Kinases.”Progress in Cell Cycle Research, 1995;1:351-363). Typical enzymes includeserine/threonine kinases such as the cyclin-dependent kinases (cdks) cdkl, cdk2,cdk4. cdk5. cdk6. as well as tyrosine kinases involved in cell cycle régulation.Increased activity or temporally abnormal activation or régulation of these kinaseshas been shown to resuit in development of human tumors and other proliférativedisorders. Compounds that inhibit cdks. either by blocking the interaction betweena cyclin and its kinase partner, or by binding to and inactivating the kinase, causeinhibition of cell prolifération, and are thus useful for treating tumors or otherabnormally proliferating cells.
Several compounds that inhibit cdks hâve demonstrated preclinicalantitumor activity. For example, flavopiridol is a flavonoid that has been shown tobe a potent inhibitor of several types of breast and lung cancer cells (Kaur et al., J. Natl. Cancer Inst.. 1992:8-4:1736-1740; Int. J. Oncol.. 1996:9:1143-1168). Thecompound has been shown to inhibit cdk2 and cdk4. Olomoucine [2-(hydroxyethylamino)-6-benzylamine-9-methylpurine] is a potent inhibitor ofcdk2 and cdk5 (Vesely et al.. Eur. J. Biochem., 1994;224:771-786). and has beenshown to inhibit prolifération of approximately 60 different human tumor cellUnes used by the National Cancer Institute (NCI) to screen for new cancer 012161 thérapies (Abraham et al.. Biology of the Cell. 1995;83:105-120). More recently.the purvalanol class of cdk inhibitors has emerged as more potent dérivatives ofolomoucine (Gray N.S. et al.. Science. 1998:281:533-538).
Tyrosine kinases are essential for the propagation of growth factor signaltransduction leading to cell cycle progression, cellular prolifération,différentiation, and migration. Tyrosine kinases include cell surface growth factorreceptor tyrosine kinases such as FGFr and PDGFr. as well as nonreceptortyrosine kinases, including c-Src and lck. Inhibition of these enzymes has beendemonstrated to cause antitumor and antiangiogenesis activity (Hambv et al.,Pharmacol. Ther.. 1999:82(2-3):169-193). • Several pyrido[2,3-d]pyrimidines that inhibit cdks and growth factor-mediated kinase enzymes are known (WO 98/33798). US Patent Nos. 5.733.913and 5.733,914 describe 6-aryI-pyrido[2.3-d]pyrimidines.
Despite the progress that has been made. the search continues for lowmolecular weight compounds that are orally bioavailable and useful for treating awide variée' of human tumors and other proliférative disorders. includingrestenosis, angiogenesis. diabetic retinopathy. psoriasis, surgical adhesions,macular degeneration. and atherosclerosis. The présent invention provides suchcompounds. their pharmaceutical formulations, and their use in treatingproliférative disorders.
SUMMARY OF THE INVENTION
This invention provides novel pyrido[2.3-d]pyrimidine-2.7-diaminecompounds which function as inhibitors of cell cycle regulatory kinases such asthe cvcîin dépendent kinases as well as the growth factor-mediated tyrosinekinases. Thus. these compounds are useful to treat cell proliférative disorders suchas atherosclerosis and restenosis: cancer: angiogenesis: viral infections includingDNA viruses such as herpes and RNA viruses such as HIV: fungal infections;type 1 diabètes, diabetic neuropathy and retinopathy: multiple sclerosis:glomerulonephritis: neurodegenerative diseases including Alzheimer's disease;autoimmune diseases such as psoriasis, rheumatoid arthritis. and lupus: organ 012161 transplant rejection and host versus graft disease: goût: polvcystic kidnev disease:and inflammation including inflammatory bowel disease.
Accordingly, the présent invention provides pyrido[2.3-d]pvrimidinehaving the generic structure of Formula I
HN wherein: R2, R7 RlS^R^andRlS are independentiv hvdrogen, or lower alkyl, lower alkenyl. or lower alkynyl. each of which is optionally substituted with up to 5 groups independently seîected from halogen. cyano. nitro, -R9, -NR^RlO. -OR9. -(CH2)nCO2R9,-(CH2)nSO2Rn, -(CH2)nRH. -COR9, -CONR9R^, -SO3R9,-SO2NR9R10, -SO2R9 -SR9. -PO3R9R10. -POR9Rl°.-po(nr9r19)2. -nr9cor’o -nr9co2rio. -nr9co\*r9rio,-NR9SO2R10. or a heterocvcie optionaliv substituted with up to 3 groups independentiv seîected from -R9. -NR9R^. -OR9.-NR9COR10. -CORlO -(CH2)nSO2RH. -(CH2)nRn. or -(CH2)nRl2 optionally substituted with up to 5 groups independentlyseîected from halogen. cyano, nitro. -R9, -NR9R1 θ. -OR9.-fCH2)nCO2R9. -fCH2)nSO2R^ L -(CH2)nR^. -COR9.-C0NR9r1°. .SO3R9. -S02NR9RlO -SO2R9. -SR9.-PO3R9R10. -POR9R19. -PO(NR9R10)2. -NR9COR10.-NR9CO2R10. -NR9CONR9r10. -NR9SO2R10. or 012161 a heterocycle optionallv substituted with up to 5 groups independentlv selected from -R9. -NR9r10. -OR9.-NR^CORiO. -COR*0. -(CH2)nSO2R5 L -(CH2)nRl 1; R- is halogen. cyano. niiro. -R9. -NR9Rl^. or -OR9: R6 is halogen. cyano. niiro. -R9. -NR9RlQ. -OR9. -CO2R9. -COR9, -CONR9R10. -NR9COR10. -SOoN^RI0. -SO2R9. -SO3R9 -SR9.-PO3R9Rl°. -POR9Rw. -PO(NR9Rl0)2. or lower alkenyi or lower alkvnvl opxionally substituted with -R9: RS is H. -CO2R1< -CORÎ3. -CONRI3R14. -CSNR13RK -C(NRî3)NR14R15,-SO3R13. -SO2R13. -SO2NR13R14. -PO3R13R14 -PORl3Rl4,-PO(NRI3R^4)2· R9 and R^ are independentlv hvdrogen. or lower alkyl. optionallv substituted with up to 3 groups selected from the group consisting of halogen. amino. mono- or dialkylamino,hydroxy. lower alkoxy. phenyl or substituted phenyl, or when taken together with the nitrogen to which they are attached, R9 and RJ θ form a ring having from 3-7 members. up to four of which
O mav be selected from p . O. S. and NR-θ. where R-θ is hvdrogen. iower alkyl. or -CO lower alkyl: R11 is a heteroarvl or a heterocyclic group:
Rl - is a cycloalkvl. a heterocyclic. an aryl. or a heteroarvl group: n is 0. I. 2. or 3: and the pharmaceutically acceptable salts. esters, amides. and prodrugs tnereof.The présent invention also provides a composition that comprises a compound of Formula 1 together with a pharmaceutically acceptable carrier,diluent, or excipient.
The présent invention also provides methods for inhibiting cyclin-dependent kinase and growth factor-mediated kinase enzymes.
V 012161 te
The présent invention also provides a method of treating subjects sufferincfrom diseases caused bv cellular prolifération. The method comprises inhibitingprolifération of tumorigenic cells of épithélial origin and vascular smooth muscleprolifération, and/or cellular migration by administering a therapeutically effectiveamount of a compound of Formula I to a subject in need of treatment.
The présent invention also pro vides a method of treating subjects sufferinsfrom diseases caused by DNA tumor viruses. such as herpes viruses comprisingadministering a compound of Formula I.
DETAILED DESCRIPTION OF THE INVENTION
The compounds encompassed by the instant invention are those describedby the general Formula I set forth above, and the pharmaceuticallv acceptablesalts. esters, amides. and prodrugs thereof.
In addition to the compounds of Formula I. the invention providespreferred compounds of Formula II:
-R'
II wherein R^. R^. R^. and R&amp; are as defined above for Formula I: R16. R17. ancj r18 are as defined above for the fCH2)nR^- substituents. andpreferably are independently hvdrogen. halogen. amino. mono- ordialkylamino. hvdroxy. lower aikyl. lower alkoxy. cyano. nitro. carboxv,carboxyalkvl. aminocarbonyl. mono- or dialkylaminocarbonyl,alkylcarbonyl. -SO3R9. -SO2NR9Rl0. -SO2R9. -SR9. -PO3R9R10, 01 2161 -6- -POR9R10. -PO(NR9R10)2· -NR9COR10. -NR^CbR10.-NR9CONR9R10. -NR9SO2R10: or
Rl6 js a carbocyclic group containing fforn 3-7 members. up to 2 of which members are heteroatoms selected from oxygen and nitrogen. wherein the 5 carbocyclic group is unsubstituted or substituted with 1. 2. or 3 croups as defined above, but preferabiy are independently selected from the groupconsisting ofhalogen. hvdroxy. lower alkyl. trifluoromethyl. lower alkoxy.amino. mono- or dialkyiamino. aryl, heteroaryl. arylalkyl. heteroarvlalkyl.heteroarylsulfonvl. heteroaryisulfonylalkvi, heterocyclylalkyl. 10 heterocyclvlsulfonyl. or heterocyclylsulfonvlalkyl.
Preferred compounds of Formula II are where R^ is hydrogen or lower alkyl; R 6 is hydrogen, lower alkyl. cvano or halogen; R1? and R^ areindependently hydrogen. halogen. amino. mono- or dialkyiamino. hvdroxy. loweralkyl. lower alkoxy, aminocarbonvl. mono- or dialkyiaminocarbonyl. 15 -SO2NR9RlQ or -NR9COR10: and R1^ is optionally substituted N-piperidine.N-piperazine. or N-pyrrolidine. for instance where the ring substituents areselected from -R9, -NR9R10, -OR9. NR9COR10. and COR10.
In addition, the présent invention also provides preferred compounds ofFormula III: R5
wherein R-, R^, and R^ are as defined above for Formula I: and R19 is hydrogen. or lower alkyl. lower alkenyl. or lower alkynyl. each of which is optionailysubstituted with up to 5 croups independently selected from il 012161
PC -7- halogen, amino. mono- or dialkylaraino. hydroxy. lower alkoxy.cyano, nitro. carboxy, carboxvalkyi. aminocarbonyl. mono- ordialkylaminocarbonyl, lower alkylcarbonyl. -SO3R9. -SO2NR9R5 °, -SO2R9. -SR9. -PO3R9R10. -POR9RlO. 5 -PO(NR9RÎ0)2. -NR9cORΰ. -NR9CO2R10. -NR9CONR9r10. -NR9S02R}0, where R9 and RIO are as defined above. or aryl. heteroaryl, arylalkyl. heteroarylalkyl. cvcloalkyl or cycloaikyl-alkvl,where each aryl. heteroaryl or cvcloalkyl group is optionallysubstituted with up to 5 groups independently selected from 10 halogen. amino. mono- or dialkvlamino, hydroxy, lower alkoxy. cyano. nitro, carboxy, carboxyalkvi. aminocarbonyl, mono- ordialkylaminocarbonyl. alkylcarbonyl. -SO3R9, -SO2NR9R^.-SO2R9, -SR9. -PO3R9R10, -POR9R10. -PO(NR9RlO)2,-NR9COR10, -NR9CO2r10 -NR9cONR9r10, -NR9sO2R10. or 15 a (CH2)n-carbocyciic group containing from 3-7 members, up to 2 of which members are heteroatoms selected from oxygen andnitrogen. wherein the carbocyclic group is unsubstituted orsubstituted with 1,2. or 3 groups independently selected from thegroup consisting of halogen. hydroxy, lower alkvl. trifluoromethyl. 20 lower alkoxy. amino. mono- or dialkyiamino. aryl. heteroaryl, arylalkyl. heteroarylalkyl. heteroarylsulfonvl.heteroarylsulfonylalkyl. heterocyclvlalkyl, heterocyclylsulfonyl, orheterocyclylsulfonylalkyl; and R-1 is hvdrogen. lower alkyl. or lower alkyl substituted with phenyl or 25 substituted phenyl.
Preferred compounds of Formula III are where R3 is hvdrogen or lower alkyl. R 6 is hvdrogen or halogen. R- is optionally substituted phenyl; R~1 ishvdrogen or methyl; and R^9 is optionally substituted lower alkyl. cvcloalkyl, or(CH2)n-carbocvciic. 012161 -8-
An especially preferred group of pyrido[2.3-d]pyrimidines hâveFormula IV:
IV wherein R A R A RlA Ri”. R^ A R^A and R-1 are as defîned above. Preferred5 compounds of Formula IV are those wherein R-1 is hvdrogen or methyl.
Another especially preferred group of invention compounds hâveFormula V:
wherein R A rA R^A Rl A Rl A R19. and R-1 are as defîned above. Preferred10 compounds of Formula V are those wherein R-1 is hvdrogen or methvl.
The most preferred invention compounds hâve Formula VI 012161
H or alkanoyl wherein RÂ RA Ri?. and Rl8 are as defmed above. and R-- and R-^independentiy are hvdrogen or alkyl.
By "alkyl.” “lower alkyl.” and "(Ci-Cio)-alkyl" in the présent invention is5 meant straight or branched chain alkyl groups having 1 to 10 carbon atoms, preferably C J-Cg alkyl. Tvpically alkyl groups include methyl, ethyl. n-propyl, isopropvl. n-butyl, sec-butyl. tert-butvl. pentyl, 2-pentyl. isopentyl. neopentyl,hexyl. 2-hexvl. 3-hexvl. decvl. octvl. and 3-methvlpentyl. These groups may besubstituted. for instance wiîh halo. C]-C3 alkyl. amino. alkylamino. dialkylamino, 10 hvdroxv. alkoxy. and the like. Examples include chloromethyl. 2-amino-ethyl. and 3-dimethyI-aminopropyl.
By "alkenyl." '‘lower alkenyl." and (C2-Cj0)-alkenvl is meant straight orbranched chain alkyl groups having 1 to 10 carbon atoms and having 1 or2 nonadjacent double bonds. Examples of alkenyls include. but are not limited to, 15 3-butenyl and 1 -methyl-3-pentenyl.
By "alkynyl." “lower alkynyl." and (C2-C]0)-alkvnyi is meant straight orbranched chain alkyl groups having 1 to 10 carbon atoms and having a triplebond. Typical alkynyl groups include 2-propynyl and l.l-dimethyi-3-butynyl.Substituted alkenyl and alkynyl groups include 4.4-dibromo-2-pentenvl and 20 3-amino-5-hexynvl.
By "alkoxy." "lower alkoxy." or "(C j -C10)-alkoxy" in the présent invention is meant straight or branched chain alkoxy groups having ! to 10 carbon atoms. such as. for example, methoxy. ethoxv. propoxy. isopropoxv n-butoxy. 012161 -10- sec-butoxy. tert-butoxy. pentoxy. 2-pentyl. isopentoxy. neopentoxv. hexoxy. 2- hexoxy, 3-hexoxy. and 3-methylpentoxy.
The term “alkanovi" means an alkyl group bonded through a carbonvlmoiety. Examples include acetvl and pentanoyl. "Aminoalkanoyl" means the alkyl 5 group is substituted with an amino group. Examples include aminoacetyl and 3- aminohexanovl. “Alkylaminoalkanoyl" means an aminoalkanoyl group whereinthe amine is substituted with a C j -C j q alkyl group. and includes methylaminoaceiyl and 4-(isobutyIamino)-octanovI. “Dialkylaminoalkanoyl"means an N.N-di-substituted aminoalkanoyl group such as 10 diisopropylaminoacetvl.
By halogen in the présent invention is meant fluorine, bromine. chlorine.and iodine.
The term "aryP means an unsubstituted aromatic carbocvclic group having 1 a single ring (e.g.. phenyl). multiple rings (e.g.. biphenyl), or multiple condensed15 rings in which at least one is aromatic (e.g.. 1.2.3.4-tetrahvdronaphthyl, naphthyl,anthryl. or phenanthryl). The term "substituted aryl" means an aryl substituted by 1 to 4 substituents selected from alkyl. O-alkyl and S-alkyl. -OH, -SH. -CN, halo, 1.3-dioxolanyl. -CF3, -NCb. -NH2. -NHCH3. -N(CH3)2» -NHCO-alkyl,-(CH2)mCO2H. -(CH2)mCO2-aIkyl. -(CH2)mSO3H. -NH alkyl, -N(alkyl)2, 20 -(CH2)mPO3H2. -(CH2)mPO3(alkyl >2- -(C^ln^NHo. and -(CH2)mSO2NH- alkyl. w'herein alkyl is deflned as above and m is 0. 1.2. or 3. Some examples ofsubstituted arvi groups are methylphenyl. isopropoxyphenvl. chiorophenyl.2-bromo-3-trifluoromethyl-4-nitro-5-aminophenyI. 4-bromobiphenyl, 3-acetamidonaphthyl. 3-dimethyiaminoanthryl. 3.4-dimethoxyphenanthryi, and 2,8- 25 dibromobiphenylen-l-yl.
By "heteroaryl" is meant one or more aromatic ring svstems of 5-, 6-. or7-members containing at least i and up to 4 heteroatoms selected from nitrogen.oxygen. or sulfur. Such heteroaryl groups include. for example, thienyl. furanyl,thiazolvL imidazolyl. (is)oxazolyl. tetrazolyl. pyridyl. pyrazinyi, pyrimidinyl, 30 pvrazolyl, (iso)quinolinyl. napthyridinyî. phthalimidyl. benzimidazoiyl, benzoxazolyl. A "substituted heteroaryl" group can be substituted with I. 2. 3, or 4 012161 -11- of lhe groups mentioned above for "substituted aryl." such as 2.3.4.6-tetrachloropvridyi and 2-methoxy-3-trifluoromethylthien-4-yl.
The terni “heterocvciic group" means a non-aromatic ring having 5-. 6-. or7-ring atoms. from 1 to 4 of which are selected from nitrogen. oxygen. or sulfur.Examples of heterocvciic groups include morpholino, piperidino. piperazino.pyrrolidinvl. and tetrahydrothienyl. Such groups can be substituted with the samegroups described above for substituted heteroaryl. A "carbocyclic group" or “cycloaîkvl" is a nonaromatic cvclic ring orfused rings having from 3- to 7-ring carbon members. Examples includecyclopropyl. cvclobutyl. and cycloheptyl. These rings may be substituted wjth oneor more of the substituent groups mentioned above for aryl, for example alkyl,halo, amino, hvdroxy. and alkoxy. Typical substituted carbocyclic groups include2-chlorocyclopropyl, 2,3-diethoxycyclopentyl, and 2.2.4,4-tetrafluorocyclohexyl.The carbocyclic group may contain 1 or 2 heteroatoms selected from oxygen,sulfur, and nitrogen. and such ring Systems are referred to as “heterocyclyl" or“heterocyclic". Examples include piperidyl, piperazinyl, pyrrolidinvl, pvranyl,tetrahydrofuranyi, and dioxanyl. These heterocyclyl groups may be substitutedwith up to 4 of the substituent groups mentioned for aryl to give groups such as 3.5-dimethylpiperazin-l -yl, 3.3-diethylpiperazin-î -y 1. 3,3,4.4-tetramethylpyrroî-idinyl. 3-chloro-2-dioxanyl. and 3.5-dihydroxymorpholino. These can also bear aketo group. for instance, 3-ketopiperidvl.
The term "cancer" includes. but is not limited to. the following tumortypes: breast, ovarv. cervix. prostate, testis. esophagus. glioblastomaneuroblastoma. stomach. skin. keratoacanthoma. lung. epidermoid carcinoma.large ceil carcinoma adenocarcinoma, bone. colon, adenoma. pancréas, thyroid.follicular carcinoma. undifferentiated carcinoma. papillary carcinoma. seminoma.melanoma. sarcoma. bladder carcinoma. liver carcinoma and biliarv passages,kidney carcinoma. myeloid disorders. lymphoid disorders. Hodgkin's. hairy cellcarcinoma. cancer of the buccal cavity and pharynx (oral), lip. longue, mouth.pharynx, small intestine, colon, rectum, large intestine, brain and central nervousSystem; and leukemia.
The compounds of Formulas 1 to VI can exist as pharmaceutically acceptable salis, esters, amides. and prodrugs. The term "pharmaceutically 0121P1
Wt J..I jS. -12- acceptabie salts. esters, amides. and prodrugs" as used herein refers to thosecarboxylate salts. amino acid addition salts. esters, amides. and prodrugs of thecompounds of the présent invention which are. within the scope of sound medicaljudgment. suitabie for use in contact with the tissues of patients without undue 5 toxicity. irritation, allergie response. and the like. commensurate with a reasonable benefiv'risk ratio, and effective for their intended use. as well as the zwitterionicforms. where possible, of the compounds of the invention. The term "salts" refersto the relativelv nontoxic, inorganic. and organic acid addition salts and base saltsof compounds of the above formulas. These salts can be prepared in situ during 10 the final isolation and purification of the compounds. or by separately reacting thepurified compound in its free base form. for example, with a suitabie organic orinorganic acid, and isolating the sait thus formed. Représentative salts include thehvdrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate. oxalate. valerate.oleate, palmitate. stéarate, laurate. borate, benzoate. lactate, phosphate, tosyiate, 15 citrate, maleate, fumarate. succinate. tartrate. naphthylate mesylate. glucoheptonate, lactobionate. and laurylsulphonate saits. and the like. When thecompound of the above formulas has one or more acidic groups. it can form a saitby reaction with a base. These salts may include cations based on the alkaii andalkaline earth metals. such as sodium, lithium, potassium, calcium, magnésium. 20 and the like. as well as inorganic bases such as ammonium, quatemarvammonium, and other amine cations including. tétraméthylammonium,tetraethylammonium. methylamine. dimethylamine. trimethylamine.triethylamine. ethylamine. and the like. Pharmaceutically acceptable salts arewell-known to those skilled in the art of médicinal chemistry. (See. for exampie. 25 Berge S.M. et ai.. “Pharmaceutical Salts.” J. Pharm. Sri.. 1977:66:1-19 which isincorporated herein by reference.)
Examples of pharmaceutically acceptable, nontoxic esters of thecompounds of this invention include Cj-C^ alkyl esters, wherein the alkyl groupis a straight or branc’ned hydrocarbon, substituted or unsubstituted. Esters aiso 30 include C5-C7 cvcloalkyl esters, as well as arylalkvl esters such as bertzyî andtriphenylmethyl. Cj-Cq Alkyl esters are preferred. such as methyl. ethyl.2.2.2-trichloroethyl. and tert-butvl. Esters of the compounds of the présent 012161
Pc> -13- invention may be prepared according to conventional methods. for example byréaction of an acid with an alcohol.
Examples of pharmaceutically acceptable amides of the compounds of thisinvention include amides derived from ammonia. primary Cj-Cg alkyl amines and 5 secondarv Cj -Cg dialkyl amines, wherein the alkyl groups are straight or branched. In the case of secondarv amines, the amine may also be in the form of a 5- or 6-membered heterocycle containing 1 nitrogen atom. Amides derived fromammonia. C} -C3 alkyl primary amines, and C j -Co dialkyl secondarv amines arepreferred. Amides of the compounds of the invention may be prepared according 10 to conventional methods well-knôwn to the médicinal chemists.
The term "prodrug" refers to compounds thaï are rapidiy transformed in vivo to yield the parent compound of the above formulae. for example, byhydrolysis in blood or stomach fluids. A thorough discussion of prodrugs isprovided by Higuchi T. and Stella V.. ‘*Pro-drugs as Novel Delivery Systems.*’ Vol. 15 14 of the A.C.S. Symposium Sériés, and in Bioreversible Carriers in Drug
Design, ed. Edward B. Roche. American Pharmaceutical Association andPergamon Press. 3 9S7. both of which are hereby incorporated by reference.
Représentative compounds of the invention are shown below in Table 1. 012161 -14-Tabie 1
7 8 9 -15- 012161
Table 1 (cont'd)
H 18 -16- 012161
Table 1 (cont'd)
-17- 012161
Table 1 (cont'd)
34 35 36
OH
PC 012161
Tabie 1 (cont’d)
38
3 HCl 43 012161 c -19-
I CH.
012161 -20-
Representative compounds of the présent invention, which areencompassed by Formula I. include. but are not iimited to. the compounds inTable 1 and their pharmaceuticallv acceptable acid or base addition salts. ester oramide analogs. and prodrugs thereof.
The compounds of the présent invention can exist in unsolvated forms aswell as solvated forms. including hvdrated forms. In general, the solvated forms.including hydrated forms. are équivalent to unsolvated forms and are intended tobe encompassed within the scope of the présent invention.
Some of the compounds of Formula I hâve one or more chiral centers. andcan thus exist as individual stereoisomers and mixtures thereof. Other compoundscan exist in more than one géométrie form. This invention includes al! optical andgéométrie isomers and forms. and mixtures thereof. Racemic mixtures ofinvention compounds are readily resolved into individual isomers by routinemethods such as chromatography. fractional crystaliization. and classicalrésolution using optically active acids and salts. The individual isomers can alsobe prepared by chiral synthesis. including chiral hydrogénations and the like usingcommercially available chiral catalvsts.
The compounds of the présent invention are usefu! for treating cancer ( forexample, leukemia. and cancer of the lung. breast. prostate, and skin such asmelanoma) and other proliférative diseases. including. but not Iimited to,psoriasis. HSV. HIV. restenosis. and atherosclerosis. To utilize a compound of theprésent invention to treat cancer, a patient having cancer is administered a 012161 P< therapeutically effective amount of a pharmaceutically acceptable compositioncomprising an invention compound. A further embodiment of this invention is a method of treating patientssuffering from diseases caused bv vascuiar smooth muscle cell prolifération.Compounds within the scope of the présent invention effectively inhibit vascuiarsmooth muscle cell prolifération and migration. The method entails inhibitingvascuiar smooth muscle prolifération, and/or migration by administering aneffective amount of a compound of Formulas I to VI to a subject in need oftreatment. "Subject" and "patient", as used herein. is a mammal such as a human.but also includes horses. cattle. sheep. and companion animais such as dogs andcats.
The compounds of the présent invention can be formulated andadministered in a wide variety of oral and parentéral dosage forms. includingtransdermal and rectal administration. It will be recognized to those skilled in theart that the following dosage forms may comprise as the active component eithera compound of Formula I or a corresponding pharmaceutically acceptable sait orsolvaté of a compound of Formula 1. A further embodiment of this invention is a pharmaceutical compositioncomprising a compound of Formulas I to VI together with a pharmaceuticallyacceptable carrier, diluent, or excipient therefor. For preparing pharmaceuticalcompositions with the compounds of the présent invention, pharmaceuticallyacceptable carriers can be either a solid or liquid. Solid form préparations includepowders. tablets, pills. capsules, cachets, suppositories. and dispensable granules.A solid carrier can be one or more substances which may also act as diiuents.flavoring agents, binders. preservatives. tablet disintegrating agents, or anencapsulating material.
In powders. the carrier is a nnely divided solid such as talc or starch whichis in a mixture with the finely divided active component. In tablets, the activecomponent is mixed with the carrier having the necessary binding properties insuitable proportions and compacted in the shape and size desired.
The formulations of this invention preferably contain from about 5% toabout 70% or more of the active compound. Suitable carriers include magnésiumcarbonate, magnésium stéarate, talc, sugar. lactose, pectin. dextrin. starch. gelatin. 012161 tragacanth. methylcellulose. sodium carboxymethylcellulose. a low meiting wax.cocoa butter, and the like. A preferred form for oral use are capsules, whichinclude the formulation of the active compound with encapsulating material as acarrier providing a capsule in which the active component with or without other 5 carriers, is surrounded bv a carrier, which is thus in association with it. Similarly. cachets and lozenges are included. Tablets, powders. capsules, pills. cachets, andlozenges can be used as soiid dosage forms suitabie for oral administration.
For preparing suppositories. a low meiting wax. such as a mixture of fattyacid glvcerides or cocoa butter, is first melted. and the active component is 10 dispersed homogeneously therein. as by stirring. The molten homogenous mixtureis then poured into convenient size molds. allowed to cool. and thereby to solidify.
Liquid form préparations include solutions, suspensions, and émulsionssuch as water or water/propylene glycol solutions. For parentéral injection, liquidpréparations can be formulated in solution in aqueous polyethylene giycol 15 solution, isotonie saline. 5% aqueous glucose, and the like. Aqueous solutions suitabie for oral use can be prepared by dissolving the active component in waterand adding suitabie colorants, flavors. stabilizing and thickening agents as desired.Aqueous suspensions suitabie for oral use can be made by dispersing the finelydivided active component in water and mixing with a viscous material. such as 20 natural or svnthetic gums. resins. methylcellulose. sodium carboxymethylcellulose. or other well-known suspending agents.
Also included are solid form préparations that are intended to beconverted. shonlv before use. to liquid form préparations for oral administration.Such liquid forms include solutions, suspensions, and émulsions. These 25 préparations mav contain. in addition to the active component. colorants, flavors, stabilizers. buffers. artificial and natural sweeteners. dispersants, thickeners.solubiiizing agents, and the like. Waxes. polymers. microparticles. and the like canbe utilized to préparé sustained-reiease dosage forms. Also. osmotic pumps can beempioyed to deiiver the active compound uniformly over a prolonged period. 30 The pharmaceutical préparations of the invention are preferably in unit dosage form. In such form. the préparation is subdivided into unit dosescontaining appropriate quantities of the active component. The unit dosage formcan be a packaged préparation, the package containing discrète quantities of 012161 -23- preparation. such as packeted tablets. capsules, and powders in vials or ampules.Also. the unit dosage form can be a capsule, tablet. cachet, or lozenge itself. or itcan be the appropriate number of any of these in packaged form.
The therapeutically effective dose of a compound of Formula I willgenerally be from about 1 to about 100 mg/kg of body weight per day. Typicaladult doses will be about 50 to about 800 mg per day. The quantity of activecomponent in a unit dose préparation may be varied or adjusted ffom about 0.1 toabout 500 mg. preferably about 0.5 to 100 mg according to the particularapplication and the potencv of the active component. The composition can. ifdesired. also contain other compatible therapeutic agents. A subject in need oftreatment with a compound of Formula I is administered a dosage of about 3 toabout 500 mg per day. either singly or in multiple doses over a 24-hour period.
The compounds of the présent invention are capable of binding to andinhibiting the activity of proteins having the ability to phosphorÿlate otherproteins, such as cdks. PDGFr. FGFr. c-Src, and EGFr. Cdks form complexes withcyclins, and these complexes phosphorylate key proteins allowing cells to proceedthrough the cell cycle (Meijer L., Progress in Cell Cycle Research.1995:1:351-363). The compounds of this invention inhibit this phosphorylationand therefore can be used as anti-proliferative agents for the treatment of cancerand/or restenosis and other proliférative diseases.
Because of their inhibitory activity against cdks and other kinases, thecompounds of the présent invention are also useful research tools for studving themechanism of action of those kinases, both in vitro and in vivo.
The préparation and use of the compounds of this invention are furtherdescribed in the following detailed example. The examples are intended toillustrate particular embodiments of the invention, and are not intended to iimit thescope of the spécification or the daims in any way. The invention compounds areprepared by synthetic méthodologies well-known to those skilled in the art oforganic chemistry. and utilize commercially available starting materials andreagents.
It may be désirable during the synthesis of an invention compound toderivatize réactivé functional groups in the molécule undergoing reacti on so as toavoid unwanted side reactions. Functional groups such as hydroxv. amino. and 012161 -24- acid groups typically are protected with suitable groups that can be readiivremoved when desired. Use of common protecting groups is described fully byGreen and Wuts in Proteciive Groups in Organic Svnthesis. John Wiley and Sons.New York. New York (2nd Edition. 1991). Tvpical hvdroxy protecting groupsinclude ether forming groups such as benzyl. and arvl groups such asze?-z-butoxycarbonyl (Boc). formyl. and acetvl. Amino protecting groups includebenzyl. arvl such as acetvl. and trialkylsilyl groups. Carboxylic acid groupstypically are protected by conversion to an ester that can be easily hydrolyzed. forexample, trichloroethyl. zerz-butyl. benzyl. and the like.
As noted above. some of the invention compounds nave one or more chiralcenters. and thus can exist as individual optical isomers and géométrie isomers,and mixtures thereof. Compound 106. for example, has two asymmetric centers.and has the cis configuration. This invention includes ail such géométrie isomers,enantiomers and RS racemates. as well as the individual R or S'isomers of chiralcompounds. Ail individual isomers and mixtures thereof are included in thisinvention. Individual isomers are readilv prepared by a chiral svnthesis. or byconventional resolution techniques well-known to those skilled in the art.
An illustration of the préparation of compounds of the présent invention isshown in Schemes 1-4. The svnthesis of Compound 1 (Example 15) is depicted inScheme 1: however. it should be recognized that the general scheme is applicableto ail of the invention compounds. Each step shown in the Schemes is furtherillustrated in the detailed examples that follow.
In Scheme 1. a 2-methylthio-4-halo-5-alkoxycarbonylpyrimidine is reactedwith ammonium hydroxide to give the corresponding 4-amino dérivative. Theester is reduced by reaction by reaction with LiA1 Hq to give the 5-hydroxymethyl analog. which in tum is oxidized to a 5-formyl dérivative. The 5-formvl group is converted to an unsaturated (acrylate) group. which is cyclizedto form a pyrido[2.3-d]pyrimidine. The pyridopvrimidine is converted to a keyintermediate. namely 2-methylsuifanyi-pyrido[2.3-d]pyrimidine-7-yiamine. whichis readily oxidized to give a 2-methvlsulfinyi analog. The 2-methylsulfinyI groupis easily displaced by reaction with an amine R2NH0 to provide the invention 01216 -25-
compounds of Formula I. The 7-amino group on the pyridopyrimidine ring isreadily converted to a urea by reaction with an isocyanate such as N=C=O 012161
012161 -27-
The reactants shown in Scheme 1 hâve the following meanings: (a) NH3: (b) LAH: (c) MnO2; (d) Ph3PCHCO2Et: (e) DBU: (f) POCI3: (g) NH3: (h) (±)-7rarcs-2-(phenyisulfonyl)-3-phenyloxaziridine: (i) 4-(4-Boc-piperidine)aniiine: (j) NaH. t-Butylisocvanate; (k) HCl 5 Scheme 2 illustrâtes an alternative synthesis of pyridopyrimidine having a urea functionality at the 7-position. Whereas such ureas were prepared inScheme 1 bv reaction of an isocyanate with a 7-amino-pyridopyrimidine,
Scheme 2 utilizes carbonyldiimidazole to provide an intermediate imidazolide.The imidazolide readilv reacts with an amine R^^NHo to give the corresponding 10 urea. Scheme 2 illustrâtes this process by showing the synthesis of Compound 51.and is more fully described in Example 32. 012161 -28-
Scheme 2
Conditions: (a) NaH. Carbonyldiimidazole: (b) Cyclopentyiamine: (c) HCl
Compounds of Formula 1 may also be prepared according to Scheme 3. 5 wherein the synthesis of compound 4 (Example 45 ) is depicted. 4-Amino-2- methanesulfanyi-pyrimidine-5-carboxaldehyde is reacted with methyl magnésiumbromide to give the corresponding 5-(2-hydroxyethyl)-pyrimidine. The alcohol isoxidized to give the methyl ketone analog. The methyl ketone is reacted withdiethvl cyanomethyl phosphonate and cyclized to a 5-methyl-7-amino- 10 pyridopyrimidine. Further reaction as in Schemes 1 or 2 gives invention compounds such as compound 4. 012161 -29-
Scheme 3
Conditions: (a) MeMgBr. (b) MnCH: (c) (EtObPfOjCH^CN
Compounds of Formula 1 may also be prepared according to Scheme 4, 5 wnerein the synthesis of compound 12 (Example 40) is depicted. in this scheme. the 2-methylthio group of a pyrimidine is First oxidized to the correspondingmethylsulfinyl analog. The methylsulfinyl group is displaced by reaction with anamine R-NFH· The 5-carboxaldehyde is then derivatized as in Scheme 1 and cyclized to give the corresponding 2-iR-NH) substituted 7-amino10 pyridopyrimidine. The 7-amino group is arylated or otherwise derivatized as illustrated in Schemes 1-3. -30-
Scheme 4 012161
Conditions: (a) (±}-Trans-2-(phenylsulfonyl)-3-phenyloxaziridine: (b) 4-(4-Boc-piperidine)-aniline: (c) (EtO)2P(O)CH2CN 5 Any of the invention compounds of Formulas 1-VI may be prepared according to Schemes 1-4. wherein the synthesis of compounds 1. 51. 4. and 12.respectively. are illustrated. Those having skill in the an of organic chemistry willrecognize that the staning materials may be varied. and additiona! steps may beemployed to produce compounds encompassed by the présent invention, as 10 demonstrated by the following spécifie examples.
The disclosures in this application of ail articles and references. including patents, are incorporated herein by reference. 012161 -31-
The invention is illustrated further by the following detaiied exampieswhich are not to be construed as limiting the invention in scope or spirit to thespécifie procedures described in them. The starting materials and variousintermediates may be obtained from commercial sources, prepared from 5 commercially available organic compounds. or prepared using well-knownsvnthetic methods. EXAMPLE 1 4-Amino~2-ntethanesulfanyl-pyrimidine-5-carboxylic acid ethyl ester
To a room température solution of 4-chloro-2-methanesulfanyl-10 pyrimidine-5-carboxylic acid ethyl ester (15.0 g, 65 mmol) in 200 mL of tetrahvdrofuran is added 25 mL of triethylamine followed by 35 mL of aqueousammonium hvdroxide. After stirring at room température for 1.5 hours, anadditional 30 mL of aqueous ammonium hvdroxide is added. and stirring iscontinued for 1 hour. The reaction mixture is concentrated in vacuo and 15 partitioned between ethyl acetate and saturated aqueous sodium bicarbonate. Theorganic laver is washed with brine. dried over magnésium sulfate, filtered. andconcentrated in vacuo. Ethyl acetate and hexane are added. and the résultant solidis collected by filtration to provide 10.84 g (79%) of 4-amino-2-methanesuifanyl-pyrimidine-5-carboxylic acid ethyl ester. 20 EXAMPLE 2 (4-Aniino-2-methanesulfanyl-pyrimidin-5-yl)-methanol A solution of 4-amino-2-methanesulfanyl-pyrimidine-5-carboxvlic acidethyl ester (13.36 g. 63 mmol) in 250 mL of tetrahvdrofuran is added dropwise toa room température suspension of lithium aluminum hvdride (3.82 g. 1 00 mmol) 25 in 250 mL of tetrahvdrofuran. After 30 minutes, the reaction is cooled to 0°C, andisopropyl alcohol is added until bubbling diminishes. The reaction is quenchedwith 15 mL of water. 15 mL of 15% NaOH. and 50 mL of water. and the mixtureis stimed for 1 hour. The white precipitate is removed by filtration and washedwith ethyl acetate. The filtrate is concentrated in vacuo and 3:1 hexane:ethyl 30 acetate is added. The soîids are collected. washed with 3:1 hexane:ethyl acetate,followed by hexane. The solid is dissolved in ethyl acetate. and the solution is 012161
PCT dried over magnésium sulfate. Filtration followed by concentration in vacuo gives8.14 g (76%) of (4-amino-2-methanesulfanyI-pyrimidin-5-yI)-methanoI.
Analysis calculated for C5H9N3OS: C. 42.09: H. 5.30: N. 24.54.
Found: C. 42.31: H. 5.24; N. 24.27. EXAMPLE 3 4- Anüno-2-inethanesulfanyl-pyrimidine-5-carboxaldehyde
To (4-amino-2-methanesuifanyl-pyrimidin-5-yl)-methanol (S. 14 g. 48 mmol) in 1 L of chloroform is added manganèse oxide (33.13 g. 381 mmol).The suspension is stirred at room température ovemight then ftltered throughcelite and washed with 300 mL of chloroform. The filtrate is concentratedin vacuo to give 8.14 g (quantitative yield) of 4-amino-2-methanesulfanyl-pyrimidine-5-carboxaIdehyde. mp 185-187°C. Literature mp = 183-184°C. JOC.1958:23:1738.
Analysis calculated for C6H7N3OS: C. 42.59; H. 4.17: N. 24.83.
Found: C. 42.84: H. 4.21: N. 24.73. EXAMPLE 4
Ethyl 3-(4-Amino-2-niethanesulfanyl-pyrimidin-5-yl)acrylate
To a room température solution of4-amino-2-methanesulfanyi-pyrimidine- 5- carboxaldeyde (4.08 g. 24.14 mmol) in 100 mL of tetrahvdrofuran is added(carbethoxymethylene) triphenylphosphorane ( 10.80 g. 31 mmol). The reactionmixture is heated at reflux for 3 hours then stirred at room température ovemight.The reaction mixture is concentrated in vacuo. and the residue is purifted by flashchromatography. eluting with 1:1 ethyl acetate:hexane. to provide 4.30 g (75%) ofethyl 3-(4-amino-2-methanesulfanyl-pyrimidin-5-yi)acrylate: mp sonens at1O8°C.
Analysis calculated for C j0H13N3O2S: C. 50.19: H. 5.48: N. 17.56.
Found. C. 50.22: H. 5.45; N. 17.24. 012161 EXAMPLE 5 2-Methanesulfanyl-8H-pyrido[2,3-dJpyrimidin-7-one
To a room température solution of ethvl 3-(4-amino-2-methanesulfanyl pyrimidin-5-yl)acryiate (368 mg. 1.53 mmol) in 3 mL of N.N- diisopropylethylamine is added 380 uL of 1.8-diazabicyclo[5.4.0Jundec-7-ene.
The reaction mixture is heated at reflux for 3 hours then cooled to roomtempérature and concentrated. The residue is purified by flash chromatographyeluting with ethvl acetate. The fractions containing the product are partiallyconcentrated in vacuo. and the solids are removed by filtration to provide 134 mg(45%) of2-methanesulfanyl-8H-pyrido[2.3-d]pyrimidin-7-one. mp 269-271°C.Analysis calculated for C8H7N3OS: C. 49.73: H. 3.65; N. 21.75.
Found: C. 49.67: H. 3.46: N. 21.49.
I EXAMPLE 6 7-Chloro-2-metltylsulfanyl-pyrido[2,3-d]pyrimidine A suspension of 1.0 g (5.2 mmol) of 2-methvlsulfanyl-8H- pyrido[2.3-d]pyrimidin-7-one (Example 5 ) in 10 mL of phosphorus oxychloride isheated under reflux for 1 hour. The resulting solution is cooled and concentratedto give a soiid. which is triturated with cold water and îiltered to give 1.05 g ofcrude product. Recrystallization from acetonitrile gives 0.76 g (69%) of theproduct. mp201-203°C. MS (APCI) M+l: Calcd 212.0: Found 212.0.
Anal. Calcd for CgHgCl j S j N3 : C. 45.39: H. 2.86: N. 19.85.
Found: C. 45.53: H. 2.90: N. 19.74. EXAMPLE 7 2-Meîhylsulfanyl-pyrido[23-djpyrimidin-7-ylaniine A suspension of2.95 g (13.9 mmol) of 7-chloro-2-methylsulfanyl- pvrido[2.3-d]pyrimidine (Example 6) in 200 mL of isopropanoi saturated withammonia is sealed and heated at 40°C for 65 hours. The suspension is resaturated 012161
PCT -34- with ammonia and heated for another 18 hours at 40°C. The solid is collected byfiltration and triturated with water to give 1.98 g (74.2%) of the product.mp >250°C. MS (APCI) M+l: Calcd 193.1; Found 193.0.
Anal. Calcd for CgHgN^Sj : C, 49.98; H. 4.19; N. 29.14.
Found: C. 50.14; H. 4.22; N. 29.04. EXAMPLE 8 2-Methanesiüfinyl~pyrido[2,3-d]pyrimidiii-7-ylanùne A suspension of 10.63 g (55.3 mmol) of 2-methylsulfanyl- pyrido[2.3-d]pyrimidin-7-yIamine (Example 7) in 300 mL of dichloromethane and300 mL of methanol is treated with 18.06 g (69.1 mmol) of (±)-trans-2-(phenylsulfonyl)-3-phenyloxaziridine and stirred ovemight. The suspension isfiltered to remove a small amount of solid. concentrated to approximately 25 mL.and diluted with ethvl acetate. The solid is collected by filtration to give 9.27 g(80.5%) of the product. mp 180°C (dec). MS (APCI) M+l: Calcd 209.0: Found 209.1. EXAMPLE 9 !\’--Phenyl-pyrido[2.3-d]pyrinüdine-2,7-diamine A suspension of 0.44 g (2.1 mmol) of 2-methanesulfmyl-pyrido[2.3- d]pyrimidin-7-ylamine (Example 8) and 0.39 mL (4.2 mmol) of aniline in 2 mL ofdimethvlsulfoxide is heated at 100°C ovemight. The resulting solution is cooledand poured into water. Ethyl acetate is added to the suspension, and the solid iscollected by filtration. The soiid is purified by flash chromatography. eluting withgradient of 0% to 20% methanol/dichioromethane during 30 minutes to give0.14 g (29%) of the product. mp 255-260°C. MS (APCI) Μ-1 : Calcd 238.1 : Found 238.1.
Anal. Calcd for C13H] jN^-O.IS H2O: C. 64.92: H. 4.76: N. 29.12.
Found: C. 65.26: H. 4.75: N. 28.76. 012161 -35- EXAMPLE 10 l-tert-Butyl-3-(2-phenylamino-pyrido[2,3-d]pyrimidin-7-yl)-urea Το a solution of 0.1022 g (0.431 mmol) of N2-phenyl-pyrido[2.3- d]pyrimidine-2,7-diamine (Example 9) in 2 mL of dimethylformamide. cooled in5 an ice bath. is added 0.019 g (0.47 mmol) of 60% sodium hvdride. The resulting solution, cooled in an ice bath. is then treated with 0.054 mL (0.47 mmol) oftert-butyl isocyanate. The solution is stirred cold for 15 minutes, then at roomtempérature for 1 hour. The solution is poured into ice-water to give a solid whichis collected by filtration and washed with hexane to give 0.0849 g (57.8%) of the 10 product (compound 45), mp 227°C (dec). MS (APCI) M+l: Calcd 337.2; Found 337.1.
Anal. Calcd for CjgFboNôOl -0.27 HoO: C, 63.35; H. 6.07; N. 24.63.
Found; C. 63.73; H. 5.82; N. 24.20. 15 EXAMPLE 11 4-(4-Nitrophenyl)-piperazine-l-carboxylic acid tert-butyl ester A suspension of 7.5 g (36 mmol) of 1 -(4-nitrophenyl)-piperazine and 6.94 mL (40 mmol) of ethyl-diisopropvl-amine in 75 mL of dichloromethane istreated with 8.69 g (40 mmol) of di-tert-butyl dicarbonate and stirred at room 20 température ovemight. The resulting solution is washed with saturated aqueous sodium bicarbonate, then with water. dried (magnésium sulfate), and concentrated.The resulting material is purified by flash chromatography eluting with a gradientof 10% to 30% ethyl acetate/hexane during 10 minutes to give 8.62 g (77.5%) ofthe product. mp 136-140°C. 25 MS (APCI) M+i: Caicd 308.2; Found 308.2. EXAMPLE 12 4-(4-Amino-phenyl)-piperazine-l-carboxylic acid tert-butyl ester
To a suspension of 1.46 g (4.8 mmol) of 4-(4-nitrophenylj-piperazine-1-carboxyiic acid tert-butyl ester (Example 11 ) and 1 g of Raney Nickel in 50 mLof tetrahydrofuran is added hvdrogen to an initial pressure of 54.5 psi. The 30 012161 PCT?. -36- reaction is shaken for 14 hours and then fiitered. The filtrate is concentrated togive 1.29 g (97%) of the product as a soiid. MS (APCI) M+l: Calcd 27S.2; Found 278.2.
Anal. Calcd for C15H23N3O2: 5 C. 64.96: H. 8.36: N. 15.15.
Found: C. 65.22; H. 8.58: N. 14.58. EXAMPLE 13 4-[4-(7-Amino-pyrido[2,3-d]pyrimidin-2-ylamino)-phenyl]-piperazine-J-carboxylic acid tert-butyl ester 10 B y substituting 4-(4-amino-phenyl)-piperazine-l-carboxylic acid tert-butyl ester (Exampie 12) for aniline in Example 9. 0.0744 g (36.0%) of the product isobtained. mp 219-220°C. MS (APCI) M+l : Calcd 422.2: Found 422.2.
Anal. Calcd for C22H27N7O2O.5 FBO: 15 C. 61.38: H. 6.56: N. 22.77.
Found: C. 61.34: H. 6.30: N. 22.47. EXAMPLE 14 4-{4-[7-(3-tert-Butyl-ureido)-pyrido[2,3-d]pyrimidin-2-ylamino]-phenyl}-piperazine-l-carboxyiic acid tert-butyl ester 20 By substituting 4-[4-(7-amino-pyrido[2.3-d]pyrimidin-2-ylamino)- phenvl]-piperazine-l-carboxylic acid tert-butyl ester (Exampie 13) for N--phenvl-pyrido[2.3-d3pyrimidine-2.7-diamine in Exampie 10. 0.3354 g (67.9%) of theproduct (compound 79) is obtained. mp 225°C <dec). MS (APCI) M+l: Calcd 521.3: Found 521.2. 25 Anal. Calcd for Cyvt-^NgC^: C. 62.29: H. 6.97: N. 21.52.
Found: C. 62.33: H. 6.81: N. 21.43. 012161 EXAMPLE 15 1- tert-Biityl-3-[2-(4-piperazin-l-yl-phenylanüno)-pyrido[2,3-d]pyrimidin-7-yl]-urea
To a suspension of 0.100 g (0.192 mmol) of 4-{4-[7-(3-tert-butyl-ureido)-5 pyrido[2.3-d]pyrimidin-2-ylamino]-phenyl}-piperazine-l-carboxylic acid tert- butyl ester (Example 14) in 2 mL of methanol is added 2 mL of 4 M hydrogenchloride/dioxane to give a solution. The suspension is stirred at room températureovemight. then diluted with diethyl ether. The material is collected by filtration togive 0.0941 g (93.4%) of the product (compound 1). mp 215°C (dec). 10 MS (APCI) M-H : Calcd 421.2: Found 421.1.
Anal. Calcd for C22H28NgO]-2.lO HCl· 1.51 H2O: C. 50.40: H. 6.37: N. 21.37; Cl (total). 14.20.
Found: C. 50.40: H. 6.18: N. 21.03: Cl (total). 14.33. t EXAMPLE 16 15 4-{4-[7-3-Cyciohexyl-ureido)-pyrido[2,3-d]pyrimidin-2-ylamino]-phenyl}-piperazine-l-carboxylic acid tert-butyl ester
By substituting cvclohexvl isocyanate for tert-butyl isocyanate inExample 14. 0.1463 g (70.4%) of the product (compound 80) is obtained.mp 241°C (dec). 20 MS (APCI) M-l: Calcd 547.3: Found 547.4.
Anal. Calcd for C29H3gNgO3-0.28 H2O: C. 63.13: H. 7.04: N. 20.31.
Found: C. 63.14: H. 6.81: N. 20.25. EXAMPLE 17 25 l-Cyclohexyl-3-l2-(4-piperaziii-l-yl-phenylamino)-pyrido[2.3-d]pyrinüdin- 7-ylf-urea
By substituting 4- 14-[7-(3-cyclohexyl-ureido)-pyrido[2.3-d]pyrimidin- 2- yiamino]-phenyl}-piperazine-î-carboxylic acid tert-butyl ester (Example 16) for4- ,f4-[7-(3-tert-butyl-ureido)-pyrido[2.3-d]pyrimidin-2-ylamino]-phenyl}- 012161 -38- piperazine-l-carboxylic acid tert-butyl ester in Example 15. 0.0871 g (81.4%) ofthe product (compound 9) is obtained. mp 200°C (dec). MS (APCI) M+l : Calcd 447.3: Found 447.3.
Anal. Calcd for C24H3oNgOi-2.55 HC1-2.82 H2O: C. 48.83: H. 6.52: N. 18.98: Cl (total), 15.31.
Found: C. 48.83: H, 6.18: N. 18.89: Ci (total). 15.37. EXAMPLE 18 N~-(4-Fluoro-3-methyl-pheny!)-pyrido[2,3-d]pyrinùdine-2,7-dicjnùne
By substituting 4-fluoro-3-methylaniline for aniline in Example 9. 0.2025 g (39.2%) of the product is obtained as a solid. MS (APCI) M+l : Calcd 270.1 : Found 270.0. EXAMPLE 19 l-tert-Butyl-3-[2-(4-fluoro-3-methyl-phenylamino)-pyrido[2.3-d]pyrimidin- 7-yl]-urea
«K
By substituting N—(4-fluoro-3-methyl-phenyl)-pyrido[2.3-d]pyrimidine- 2.7-diamine (Example 18) for N--phenyI-pyrido[2.3-d]pyrimidine-2.7-diamine inExample 10. 0.0656 g (47.9%) ofthe product (compound 46) is obtained.mp 230°C (dec). MS (APCI) M-l: Calcd 369.2: Found 369.1.
Anal. Calcd for C j çH2 ] F ] NgO ] : C. 61.94: H. 5.75: N. 22.81.
Found: C. 61.82: H. 5.73: N. 22.75. EXAMPLE 20 1 -(4-Chloro-phenyl)-3-[2-(4-fluoro~3-ntetltyl-phenylaniino)- pyrido[2.3~d]pyrimidin-7-yll-urea
By substituting 4-chlorophenvl isocyanate for tertiary-butvl isocyanate inExample 19. 0.050 g (37%) of the product (compound 47) is obtained.mp >250°C. MS (APCI) M+l: Calcd 423.1: Found 423.1. -39- 012161
Anal. Calcd for C2iHj6FîC1jN6Oi-0.23 FBO: C. 59.07: H. 3.89: N. 19.68.
Found: C. 59.09: H. 3.97: N. 19.65. EXAMPLE 21 l-{2-[4-(4-Acetyl-piperazin-l-yl)-phenylamino]-pyrido[2,3-d]pyrimidin- 7-yl}-3-tert-butyl-urea
To a suspension of 0.145 g (0.277 mmol) of l-ten-butyi-3-[2-(4-piperazin-l-yl-phenylamino)-pyrido[2.3-d]pyrimidin-7-yl]-urea (Example 15) in 5 mL ofdichloromethane is added 0.19 mL (1.11 mmol) of ethyl-diisopropyl-amine. Thesuspension is cooled in an ice bath and treated with 0.024 mL (0.33 mmol) ofacetvl chloride. The suspension is stirred at room température ovemight, thenfiltered. The solid is washed with dichloromethane. The filtrate and washings arecombined, washed with water. dried (magnésium sulfate), and concentrated. Thematerial is purified by flash chromatography eluting with a gradient of 0% to 5%methanol/dichloromethane during 30 minutes to give 0.0674 g (51.8%) of theproduct (compound 5). mp 206-208°C (dec). MS (APCI) M+l : Calcd 463.3: Found 463.3.
Anal. Calcd for C24H3oNg02-0.40 H2O: C. 61.36: H. 6.61: N. 23.85.
Found: C. 61.38: H. 6.37: N. 23.98. EXAMPLE 22 4-{4-[7-(3~Isopropyl-ureido)-pyrido[2.3-d]pyrimidin-2-ylamino]-phenyl}-piperazine-l-carboxylic acid tert-butyl ester
By substituting isopropyl isocyanate for tert-butyl isocyanate inExample 14. 0.909 g (69.9%) of the product is obtained as a solid. MS (APCI) M-H : Calcd 507.3: Found 507.4. -4°- EXAMPLE 23 J-Isopropyl-3-[2-(4-piperaziit-l-yl~pltenylamino)-pyrido[2,3-d]pyrimidin-7-ylJ- urea
By substituting 4-{4-[7-(3-isopropyl-ureido)-pyrido[2.3-d]pyrimidin-2-ylamino]-phenyl}-piperazine-l-carboxyiic acid tert-butyl ester (Example 22) for4-{4-[7-(3-tert-butyl-ureido)-pyrido[2.3-d]pyrimidin-2-ylamino]-phenyl}-piperazine-l-carboxylic acid tert-butyl ester in Example 15. 0.0287 g (27.9%) ofthe product (compound 48) is obtained. mp 190°C (dec). MS (APCI) M+l: Calcd 407.2: Found 407.1.
Anal. Calcd for C2iH2^NgOp2.05 TFA-0.84 FbO: C. 46.00: H. 4.57: N. 17.10.
Found: C. 46.00: H. 4.65: N. 17.09. EXAMPLE 24
Cis-3,5-dimethyl-l-(4-nitro-phenyl)-piperazine A suspension of 6.74 g (47.8 mmol) of 4-fluoro-nitro-benzene and 10.91 g(95.5 mmol) of cis-2.6-dimethyl-piperazine is heated at 45°C for 1 hour. Thereaction mixture is cooled and shaken with dichloromethane and water. Theorganic laver is dried (magnésium sulfate) and concentrated to give 11.62 g(>100%) of the product as a solid.
EXAMPLE 25
Cis-2.6-dimetliyl-4-(4-ititro-plienyl)-piperaziue-l-carboxylic acid tert-butyl ester
By substituting cis-3.5-dimethyl-l-(4-nitro-phenyl)-piperazine (Example 24) for l-(4-nitrophenvl)-piperazine in Example 11. 14.87 g (92.8%) ofthe product as a solid is obtained. EXAMPLE 26 4-(4-Amino-phenyl)-cis-2.6-dimetltyl-piperazine-I-carboxylic acid tert-butylester
By substituting cis-2.6-dimethyl-4-(4-nitro-phenyi (-piperazine-1-carboxylic acid tert-butyl ester (Example 25) for 4-(4-nitro-phenyl)-piperazine- 012161 1-carboxylic acid tert-butyi ester in Example 12. 5.03 g (64.7%) of the product asa soiid is obtained. EXAMPLE 27 4-[4-(7-Amino-pyrido[2,3-d]pyriinidin-2-ylamino)-phenyl]-cis-2.6-diinetliyl-5 piperazine-l-carboxylic acid tert-butyi ester
By substituons 4-(4-amino-phenyI)-cis-2.6-dimethyl-piperazine-1-carboxyiic acid tert-butyi ester (Example 26) for aniline in Example 9. 0.6463 g(59.8%) of the product is obtained. mp 245°C (dec). MS (APCI) M+l: Calcd 450.3: Found 450.3. 10 EXAMPLE 28 4-{4-l7-(3-tert-Butyl-ureido)-pyrido[2,3-d]pyritnidin-2-ylamino]-pitenyl}-cis- 2,6-dimethyl-piperazine-l-carboxylic acid tert-butyi ester ,
By substituting 4-[4-(7-amino-pyrido[2.3-d3pyrimidin-2-yiamino)-phenyl]-cis-2.6-dimethvl-piperazine- 1-carboxylic acid tert-butyi ester 15 (Example 27) for N2-phenyl-pyrido[2.3-d]pyrimidine-2,7-diamine in Example 10,0.1828 g (74.9%) of the product is obtained as a soiid. MS (APCI) M+l : Calcd 549.3: Found 549.4. EXAMPLE 29 1- tert-Butyl-3-{2-[4-(cis-3,5-dimethyl-piperazin-l-yl)-phenylaminoJ- 2 0 pyrido[2,3-d]pyrimidin - 7-yty-urea
By substituting 4-{4-[7-(3-tert-butyl-ureido)-pyrido[2.3-d]pyrimidin- 2- ylamino]-phenyl}-cis-2.6-dimethyl-piperazine-1-carboxylic acid tert-butyi ester(Example 28) for 4-{4-[7-(3-tert-butyl-ureido)-pyrido[2.3-d]pyrimidin-2-ylamino]-phenyl}-piperazine-l-carboxylic acid tert-butyi ester in Example 15 is 25 obtained 0.0910 g (92.9%) of the product (compound 49). mp 245°C (dec). MS (APCI) M~l: Calcd 449.3: Found 449.2. PC, -42. 012161 EXAMPLE 30 4-{4-[7-(3-Cyclohexyl-ureido)-pyrido[2,3-d]pyritnidin-2-ylanünol-phenyl}-cis- 2,6-dimethyl-piperazine-l-carboxylic acid tert-butyl ester
By substituting cyciohexyl isocyanate for tert-butyl isocyanate inExample 28. 0.1156 g (60.8%) of the product is obtained as a solid. MS (APCI) M+l: Calcd 575.3; Found 575.3. EXAMPLE 31 1- Cyclohexyl-3-{2-[4-(cis-3,5-diniethyl-piperazin-l-yl)-plteHylaniinoJ-pyridoI2,3-d]pyrimidin-7-yl}-urea
By substituting 4-{4-[7-(3-cyclohexyl-ureido)-pyrido[2.3-d]pvrimidin- 2- ylamino]-phenyl}-cis-2.6-dimethyl-piperazine-l-carboxylic acid tert-butyl ester(Example 30) for 4-{4-[7-(3-tert-butyl-ureido)-pyrido[2.3-d]pyrimidin-2-vlamino]-phenyl}-piperazine-l-carboxylic acid tert-butyl ester in Example 15.0.1022 g of the product is obtained (compound 50). mp 228®C (dec). MS (APCI) M+l : Calcd 475.2: Found 475.2. EXAMPLE 32 4-{4-[7-(3-Cyclopentyl-ureido)-pyrido[2,3-d]pyrimidin-2-ylamino]-phenyl}-piperazine-l-carboxylic acid tert-butyl ester
To a solution of 0.150 g (0.36 mmol) of 4-[4-(7-amino-pyrido[2.3-d]pyrimidin-2-yiamino)-phenyl]-piperazine-l-carboxylic acid tert-butyl ester(Example 13) in 2 mL of dimethvlformamide. cooled in an ice bath. is added0.022 g (0.54 mmol) of 60% sodium hydride. The cooled solution is stirred for15 minutes, then treated with 0.088 g (0.54 mmol) of carbonyldiimidazole. Thecooled solution is stirred for another 30 minutes, then treated with 0.071 mL(0.72 mmol) of cvclopentylamine. The resulting solution is stirred at roomtempérature for 1 hour. then added to cold water. The solid is coüected byfiltration to give a first crop of material. The aqueous filtrate is then extracted withdichloromethane. and the extracts are dried (magnésium sulfate) and concentratedto give a second crop of material. The 2 crops are combined and purified by flashchromatography. eluting with a gradient of 0% to 5% methanol/dichioromethaneduring 30 minutes to give 0.1159 g (60.4%) of the product as a solid. 012161 -43- MS (APCI) M+l: Calcd 533.3: Found 533.4. EXAMPLE 33 1- Cyclopeniy'I-3-[2-(4-pipera-in-l-yI-pIieiiy!aniino)-pyrido[2.3-djpyrimidiii-7-ylJ-urea
Bv substituting 4-{4-[7-(3-cyclopentyl-ureido)-pyrido[2.3-d]pyrimidin- 2- ylamino]-phenvl}-piperazine-l-carboxylic acid tert-butyl ester (Example 32) for 4- {4-[7-(3-tert-butyl-ureido)-pyrido[2.3-d]pyrimidin-2-ylamino]-phenyl}-piperazine-I-carboxylic acid tert-butvl ester in Example 15, 0.0937 g (80.8%) ofthe producî (compound 51) is obtained. mp 210-213°C (dec). MS (APCI) M~1 : Calcd 433.2: Found 433.2.
Anal. Cale for CosHbgNgOj-2.49 HCI-1.65 HbO-O.l dioxane: C. 50.02: H. 6.21: N. 19.94: Cl (total), 15.71.
Found: C. 49.89: H. 5.81: N. 19.74: Cl (total). 14.74. EXAMPLE 34 I-(4-Amino-2-niethylsulfany{-pyriniidiii-5-yb-ethanol
To a suspension of 5.0 g (29 mmol) 4-amino-2-methyîsulfanyl-pyrimidine- 5- carboxaldehvde (Example 3) in 150 mL of tetrahvdrofuran. cooled by anice bath. is added during 20 minutes. 23.2 mL of a 3.0 M méthylmagnésiumbromide solution in diethvl ether (69.4 mmol). After 1 hour at 0°C. another23.2 mL of the 3.0 M méthylmagnésium bromide solution is added. and thesuspension is allowed to corne to room température and stirred ovemight. Thereaction is quenched with 100 mL of saturated aqueous ammonium chloride. andpartitioned between water and ethvl acetate. The organic laver is dried(magnésium sulfate) and concentrated to give 5.24 g (96%) of the product. mp 140-142°C. MS (APCI) M-î : Calcd 186.1: Found 185.9. EXAMPLE 35 l-(4-Amino-2-metliy}sulfaijyi-pyrimidin-5~yl)-ethanone
By substituting 1 -(4-amino-2-methyIsulfanyi-pyrimidin-5-yl)-ethanoi (Example 34) for (4-amino-2-methylsulfanyl-pyrimidin-5-yl)-methanol in 012161 PCT/Ifc 4 4 —Pt*
Exampie 3 and conducting the reaction at 80°C in toluene. 3.74 g (72%) of theproduct as a solid is obtained. MS (APCI) M+l: Calcd 184.0: Found 183.9. EXAMPLE 36 l-(4-Amino-2-methanesulfinyl-pyrimidiH-5-yl)-ethanone
By substituting l-(4-amino-2-methylsulfanyl-pyrimidin-5-yl )-ethanone(Example 35) for 2-methylsulfanyl-pyrido[2.3-d]pyrimidin-7-ylamine inExample 8. 9.57 g (88%) of the product as a solid is obtained. MS (APCI) M-l : Calcd 200: Found 200. EXAMPLE 37 4-[4-(5-Acety!l-4-ainino-pyrimidin-2-ylamino)-pltenyl]-pipsrazine-l-carboxylicacid tert-butyl ester t
By substituting 1 -(4-amino-2-methanesulfinyl-pyrimidin-5-yl)-ethanone(Example 36) for 2-methanesulftnyl-pyrido[2.3-d]pyrimidin-7-ylamine inExample 13. 4.04 g (65%) of the product as a solid is obtained. MS (APCI) M-l: Calcd 413: Found 413. EXAMPLE 38 4-{4-(7-Atnino-5-niethyl-pyrido{23-d]pyriniidiii-2-ylamhio)-phenyl]-piperazine-1-carboxyiic acid tert-butyl ester
To a suspension of 0.58 g (14.6 mmol, of 60% sodium hvdride in 10 mL oftetrahvdrofuran. at 0°C. is added dropwise 2.58 g (14.56 mmol) of diethyl(cvanomethyl) phosphonate. The reaction mixture is stirred at 0°C for 5 minutes,then at room température for 20 minutes. The mixture is then cooled to 0°C andtreated with 2 g (4.85 mmol) of4-[4-(5-acetyl-4-amino-pyrimidin-2-yiamino>-phenyl]-piperazine-l-carboxylic acid tert-butvi ester (Example 37). The mixture isstirred at room température ovemight. and then treated with water and saturatedaqueous ammonium chioride. The resulting solid is collected by filtration andwashed with ether to cive 1.069 g (80%) of the product. MS (APCI) M-l: Calcd 436: Found 436. 012161 wr -45- EXAMPLE 39 4'{4-[7-(3-Cyclohexyl-ureido)-5-tnetliyl-pyrido[2.3-d]pyrimidin-2-ylatiüno]-phenyl}-piperazirte-l-carboxylic acid tert-butyl ester
Bv substituting 4-[4-(7-amino-5-methyl-pyrido[2.3-d]pyrimidin-2-5 ylamino)-phenyl]-piperazine-l-carboxvlic acid tert-butyl ester (Example 38) for 4-[4-(7-amino-pyrido[2.3-d]pyrimidin-2-ylamino)-phenyl]-piperazine-l-carboxylic acid tert-butyl ester in Example 16. 0.199 g (42%) of the product as asolid is obtained. MS (APCI) M-H: Calcd 561: Found 561. 10 EXAMPLE 40 1- Cyclohexyl-3-[5-methyl-2-(4-piperazin-l-yl-phenylamino)-pyrido[2,3-d]pyrimidin-7-yl]-urea
By substituting 4-{4-[7-(3-cyclohexyl-ureido)-5-methyî-'pyrido[2.3-d)pyrimidine-2-ylamino]-phenyl}piperazine-î-carboxvlic acid tert-butyl ester 15 (Example 39) for 4-{4-[7-(3-tert-butyl-ureido)-pyrido[2,3-d]pyrimidin-2- vlamino]-phenyl}-piperazine-l-carboxvlic acid tert-butyl ester in Example 15. theproduct (compound 12) as a solid is obtained. mp 238°C (dec). MS (APCI) M+l: Calcd 461 : Found 461. EXAMPLE 41 20 5-Methyl-2-methylsulfanyl-pyrido[2,3-dJpyrimidiH-7-ylamine
By substituting 1 -(4-amino-2-methylsulfanyI-pyrimidin-5-yl)-ethanone (Example 35) for 4-[4-(5-acetyl-4-amino-pyrimidin-2-ylamino)-phenyl]-piperazine-1-carboxvlic acid tert-butyl ester in Example 38. 0.97 g (85%) of theproduct as a solid is obtained. 25 MS (APCI) M-l : Calcd 207: Found 207. EXAMPLE 42 2- Methanesulfînyl-5-methyl-pyrido[2.3-d]pyrimidin-7-yiamine
By substituting 5-methyî-2-methylsulfanyl-pyrido[2.3-d]pyrimidin-7-ylamine (Example 41 ) for 2-methylsuifanyl-pyrido[2.3-d]pyrimidin-7-viamine in 30 Example 8. 0.85 g (83%) of the product as a solid is obtained. 01216-1 -46- MS (APCI) M+l: Calcd 223; Found 223. EXAMPLE 43 4-[4-(7-Antino-5-methyl-pyrido[2,3-dJpyrimidin-2-ylamino)-phenylJ-piperazine-1-carboxylic acid tert-butyl ester 5 By substituting 2-methanesulfinyl-5-methyl-pyrido[2.3-d]pyrimidin-7- vlamine (Example 42) for 2-methanesulfinyl-pyrido[2.3-d]pyrimidin-7-ylamine inExample 13. 0.33 g (20%) of the product as a solid is obtained. MS (APCI) M+l: Calcd 436: Found 436. EXAMPLE 44 10 4-{4-[7-(3-tert-Butyl-ureido)-5-tnetliyl-pyrido[2,3-d]pyrimidiii-2-ylamino]-phenyl}-piperazine-l-carboxylic acid tert-butyl ester
By substituting 4-[4-(7-amino-5-methyl-pyrido[2.3-d]pyrimidin-2-ylamino)-phenyl]-piperazine-1-carboxylic acid tert-butyl ester (Example 43) forN--phenyI-pyrido[2.3-d]pyrimidine-2.7-diamine in Example 10. 0.17 g (45%) of 15 the product as a solid is obtained. MS (APCI) M+l: Calcd 535; Found 535. EXAMPLE 45 l-tert-Butyl-3-[5-methyl-2-(4-piperazin-l-yl-phenylamino)- pyrido[2,3-d]pyritnidin-7-yl]-urea 20 By substituting 4-{4-[7-(3-tert-butyl-ureido)-5-methyl-pyrido[2.3-
d]pyrimidin-2-ylamino]-phenyl}-piperazine-l-carboxvlic acid tert-butyl ester(Example 44) for 4-{4-[7-(3-tert-butyl-ureido)-pyrido[2.3-d]pyrimidin-2-ylamino]-phenyl}-piperazine-1-carboxylic acid tert-butyl ester in Example 15,0.070 g (72%) of the product (compound 4) as a solid is obtained. mp 230-232°C 25 (dec). MS (APCI) M-l : Calcd 435: Found 435. 012161 -47- EXAMPLE 46 6- Fluoro-2-methylsulfany[-8H-pyrido[2,3-djpyrimidin-7-one A solution of 1.74 g (10.33 mmol) of (diethoxy-phosphorvl)-fluoro-aceticacid ethyl ester in 20 mL of tetrahydrofuran is cooled to -78°C and treateddropwise with 12.9 mL (20.65 mmol) of a 1.6 M solution of n-butyl lithium inhexanes. After stirring for 30 minutes at -78°C. the solution is treated with 1.74 g(10.33 mmol) of 4-amino-2-methylsuifanyl-pyrimidine-5-carboxaldehyde(Example 3). allowed to warm to room température, and stirred ovemight. Thereaction is treated with saturated aqueous ammonium chloride. then water. Thesoiid is collected bv filtration and washed with diethyl ether to give 2.01 g (92%)of the product. MS (APCÏ) M+l: Calcd 212: Found 212. EXAMPLE 47 7- Chloro-6-fluoro-2-methylsulfanyl-pyrido[2,3-d]pyrimidine
By substituting 6-fluoro-2-methylsulfanyl-8H-pyrido[2.3-d]pyrimidin-7-one (Example 46) for 2-methylsulfanyl-8H-pyrido[2.3-d]pyrimidin-7-one inExample 6 is obtained 1.86 g (85%) the product as a solid. MS (APCI) M-l : Calcd 230.232; Found 230. 232. EXAMPLE 48 6-Fluoro-2-methylsulfanyl-pyrido[2,3-d]pyrimidine-7-ylamine
By substituting 7-chloro-6-fluoro-2-methylsulfanyî-pyrido[2.3-d]pyrimidine (Example 47) for 7-chloro-2-methylsulfanyl-pyrido[2.3-djpvrimidine in Example 7 is obtained 0.29 g (90%) of the product as a solid. MS (APCI) M+l : Calcd 211 : Found 211. EXAMPLE 49 6-Fluoro-2-methanesulfinyl~pyrido[2,3-d]pyrimi(iin-7-ylamine
By substituting 6-fluoro-2-methylsulfanyl-pyrido[2.3-d]pyrimidin-7-vlamine (Example 48) for 2-methylsulfanyl-pyrido[2.3-d]pyrimidin-7-yIamine inExample 8. 0.26 g (95%) of the product as a solid is obtained. MS (APCI) M-l: Calcd 227: Found 227.
PC 012161 EXAMPLE 50 4-[4-(7-Amino-6-fluoro-pyrido[2,3-d)pyrimiditi-2-yiamino)-phenyl]-cis- 2,6-dimethyl-piperazine-l-carboxylic acid tert-butyl ester
Bv substiiuting 6-fluoro-2-methanesuIfinyl-pyrido[2.3-d]pyrimidin-7-ylamine (Example 49) for 2-methanesulfinyl-pyrido[2.3-d]pyrimidin-7-ylamine inExample 27. 0.040 g (63%) of the product as a solid is obtained. MS (APCI) M-H: Calcd 468: Found 468. EXAMPLE 51 4-{4-[7-(3-Cyclohexyl-ureido)-6-fluoro-pyrido[2,3-d]pyriiniditt-2-ylainino]-phenyl}-cis-2.6-dimetliyl-piperazine-l-carboxyHc acid tert-butyl ester
Bv substituting 4-[4-(7-amino-6-fluoro-pyrido[2.3-d]pyrimidin-2-ylamino)-phenyl]-cis-2.6-dimethyl-piperazine-l-carboxylic acid tert-butyl ester(Example 50) for 4-[4-(7-amino-pyrido[2.3-d]pyrimidin-2-ylamino)-phenyl]-piperazine-l-carboxylic acid tert-butyl ester in Example 16. 0.10 g (74%) of theproduct as a solid is obtained. MS (APCI) M+l : Calcd 593: Found 593. EXAMPLE 52 l-Cyclohexyl-3-{2-[4-(cis-3.5-dimetliyl-piperazin-l-yl)-phenylamiiio}-6-fluoro- pyrido[2.3-djpyrimidin-~-yl/-urea
Bv substiiuting 4-{4-[7-(3-cycIohexyl-ureido)-6-fluoro-pyrido[2.3-d]pyrimidin-2-ylamino]-phenyl }-cis-2.6-dimethyl-piperazine-1 -carboxvlie acidtert-butyl ester (Example 51 ) for 4-(4-[7-(3-tert-butyl-ureido)-pyrido[2.3-d]pyrimidin-2-vlamino]-phenyl}-piperazine-l-carboxylic acid tert-butyl ester inExample 15. 0.060 g (75%) of the product (compound 52) as a solid is obtained.mp 227-229°C. MS (APCI) M+l: Calcd 493: Found 493. MU' -49- EXAMPLE 53 01 2161 4-{4-[7-(3-Cyclopentyi-ureido)-5-methyl-pyrido[2,3-d]pyrimidin-2-ylamino]-phenyl}-piperazine-l-carboxyUc acid tert-butyi ester
By substituting 4-[4-(7-amino-5-methyI-pyrido[2.3-d]pyrimidin-2-5 ylamino)-phenyl]-piperazine-l-carboxylic acid tert-’outyl ester (Example 43) for 4-[4-(7-amino-pyrido[2.3-d]pyrimidin-2-ylamino)-phenyl]-piperazine-l-carboxvlic acid tert-butyi ester in Exampie 32. 0.18 g (55%) of the product as asolid is obtained. MS (APCI) M+î: Calcd 547: Found 547. 10 EXAMPLE 54 l-Cyclopentyl-3-[5-metliyl-2-(4-piperazin-l-yl-phenylamino)- pyrido[2,3-d]pyrimidin-7-yt]-urea
By substituiing 4- {4-[7-(3-cyciopentyl-ureido)-5-methyk-pyrido[2.3-d]pyrimidin-2-yianiino]-phenyl}-piperazine-l-carboxyiic acid ten-butvl ester 15 (Example 53) for 4-{4-[7-(3-tert-butyl-ureido)-pyrido[2.3-d]pyrimidin-2- ylamino]-phenyl}-piperazine-l-carboxylic acid tert-butyi ester in Example 15,0.08 g (70%) of the product (compound 53) is obtained. mp 234°C (dec). MS (APCI) M-H: Calcd 447: Found 447. EXAMPLE 55 20 4-(4-{7-[3-(3-Hydroxy-propyl)-ureido]-pyrido[2,3-d]pyrimidin-2-ylanüno}-phenyl)-piperazine-I-carboxyttc acid tert-burvl ester
By substituting 3-amino-l-propanol for cyciopentylamine. and sodiumtertiary butoxide for sodium hvdride in Example 32. 0.1295 g (52.2%) of theproduct as a solid is obtained. 25 MS (APCI) M+l: Calcd 523.3; Found 523.2. EXAMPLE 56 l-(3-Hydroxy-propyi)-3-[2-(4-piperazin-l-yl-phenylamino)-pyrido [2,3-d]pyrimidin-7-ylJ-urea B\ substituting the product of Example 55 in Example 15. 0.1077 g of the30 product (compound 81) as a solid is obtained. mp i 83°C (dec). -50- MS (APCI) M+l: Calcd 423.2: Found 423.1. EXAMPLE 57 4-{4-{7-(3-Cyclohexyl-3-tnetltyl-ureido)-pyrido[2,3-d]pyrimidin-2-ylaniino}-phenyl}-piperazine-l-carboxylic acid tert-butyl ester 5 By substituting N-methvlcyclohexylamine for 3-amino-1 -propanol in
Example 55. 0.1932 g (72.7%) of the product as a solid is obtained. MS (APCÏ) M+l : Calcd 561.3: Found 561.2. EXAMPLE 58 l-Cyclohexyl-l-methyl-3-[2-(4-piperazin-l-yl-phenylatnino)-pyrido10 [2,3-d]pyrimidin-7-yl]-urea
By substituting the product of Exampie 57 in Example 15. 0.1645 g of theproduct (compound 65) as a solid is o'otained. mp 177°C (dec).tMS (APCI) M+l: Calcd 461.3: Found 461.2. EXAMPLE 59 15 4-(4-{7-[3-((S)-l-Hydroxymethyl-3-methyl-butyl)-ureido]-pyrido [2,3-d]pyrimidiH-2-ylamino}-phenyl)-piperazine-l-carboxylic acid tert-butylester
By substituting (S)-(+)-leucinol for 3-amino-1-propanol in Exampie 55. 0.1048 g (39.1 %) of the product as a solid is obtained. 20 MS (APCI) M+l : Calcd 565.3: Found 565.3. EXAMPLE 60 l-((S)-l-Hydroxymethyl-3-methyl-butyl)-3-{2-(4-piperazin-l-yl-phenylamino)- pyrido[2,3-d]pyrimidin-7-yl]-urea
By substituting the product of Exampie 59 in Example 15. 0.0802 g of the 25 product (compound 83) as a solid is obtained. mp 185°C (dec). MS (APCI) M+l : Calcd 465.3: Found 465.2.
012161 -51- EXAMPLE61 4-[4-(7-{[l-(4-Methyl-piperazin-l-yl)-tnethanoyl]-amino}-pyrido[2,3-d]pyrimidin-2-ylamino)-phenyl]-piperazine-l-carboxyUc acid tert-butyl ester
Bv substituting N-methylpiperazine for 3-amino-l-propanol in5 Example 55. the product as a solid is obtained. MS (APCI) M+l: Calcd 548.3: Found 548.3. EXAMPLE 62 4-Methy[-piperazine-l-carboxylic acid [2-(4-piperazin-l-yl-phettylamino)-pyrido[2,3-d]pyrimidin- 7-ylJ-amide 10 By substituting the product of Example 61 in Example 15. 0.1194 g of the product (compound 84) as a solid is obtained. mp 200°C (dec). MS (APCI) M+l: Calcd 448.3: Found 448.2.
I EXAMPLE 63 4-(4-{7-[(l-Morpholin-4-yl-methanoyl)-amino]-pyrido[2,3-d]pyrimidin-2- 15 ylamino}-phenyl)-piperazine-l-carboxylic acid tert-butyl ester
By substituting morphoiine for 3-amino-l-propanol in Example 55. the product as a solid is obtained. MS (APCI) M+l: Calcd 535.3: Found 535.2. EXAMPLE 64 20 MorphoUne-4-carboxylic acid [2-(4-piperaziu-l-yl-phenylatnino)-pyridi)J2,3-djpyrimidin-7-yl]-amide
By substituting the product of Example 63 in Example 15. 0.1132 g of theproduct (compound 85) as a solid is obtained. mp 190°C (dec). MS (APCI) M-l: Calcd 435.2: Found 435.2. 25 EXAMPLE 65 4-{4-[7-(3,3-Dipropyi-ureido)-pyrido[2,3-d)pyrimidin-2-ylamino]-phenyl}-piperazine-l-carboxylic acid tert-butyl ester
By substituting dipropylamine for 3-amino-l-propanol in Example 55. theproduct as a solid is obtained. 012161 MS (APCI) M-l: Calcd 549.3: Found 549.3. EXAMPLE 66 3- [2-(4-Piperazin-l-yl-pbenylamiito)-pyrido[2,3-d]pyrimidin-7-yl}-l,l-dipropylurea 5 By substituting the product of Example 65 in Example 15. 0.1278 g of the product (compound 86) as a solid is obtained. mp 190°C (dec). MS (APCI) M-H : Calcd 449.3: Found 449.2. EXAMPLE 67 4- [4-(7-{[l-(4-Boc-piperazin-l-yl)-niethanoyl]-amino}-pyrido[2.3-d]pyrimidin-2- 10 ylamino)-phenyl]-piperazine-l-carboxylic acid tert-butyl ester
By substituting Boc-piperazine for 3-amino-l-propanol in Example 55. theproduct as a solid is obtained. v MS (APCI) M-H: Calcd 634.3: Found 634.3. EXAMPLE 68 15 Piperazine-I-carboxylic acid [2-(4-piperazin-l-yl-phenylamino)-pyrido[2,3-dIpyrimidin- 7-ylJ-amide
By substituting the product of Example 67 in Example 15. 0.0342 g of theproduct (compound 87) as a solid is obtained. mp 220°C (dec). MS (APCI) M*1 : Calcd 434.2: Found 434.2. 20 EXAMPLE 69 4-(4-{7-[3-((R)-l-Hydroxymethyl-2-methyl-propyl)-ureido]-pyrido[2,3-d]pyrimidin-2-ylamino}-phenyl)-piperazine-l-carboxylic acid tert-butyl ester
By substituting (R)-valinol for 3-amino-l-propanol in Example 55. theproduct as a solid is obtained. 25 MS (APCI) M-H: Calcd 551.3: Found 551.3.
W pC 012161 EXAMPLE 70 l-((R)-l-Hydraxymethyl-2-methyl-propyl)-3-{2-(4-piperazin-l-yl-phenylamino)- pyrido[2,3-d]pyrimidin-7-yl]-urea
Bv substituting the product of Example 69 in Example 15. 0.0639 g of the5 product (compound 88) as a solid is obtained. mp 200°C (dec). MS (APCI) M+1 : Calcd 451.3: Found 451.2. EXAMPLE 71 4-(4-{7-[3,3-Bis-(2-hydroxy-ethyl)-ureido]-pyrido[2,3-d]pyrinüdin-2-ylamino}-phenyl)-piperazine~l-carboxylic acid tert-butyl ester 10 Bv substituting diethanoiamine for 3-amino-i-propanol in Exampie 55. the product as a solid is obtained. MS (APCI) M+l: Calcd 553.3: Found 553.2. EXAMPLE 72 l,l-Bis-(2-hydroxy-ethyl)-3-[2-(4-piperazin-l-yl-phenylanûno)-pyrido[2,3- 15 djpyrimidin-7~y!]-urea
By substituting the product of Example 71 in Example 15. 0.0916 g of theproduct (compound 89) as a solid is obtained. mp 185°C (dec). MS (APCI) M+l: Calcd 453.2; Found 453.2. EXAMPLE 73 20 6-Bromo-2-methylsulfanyl-8H-pyrido[2,3-d]pyrimidin-7-one
To 5.00 g (25.9 mmol) of 2-methanesulfanyl-8H-pyrido[2.3-d]pyrimidin-7-one (Example 5) in 130 mL of DMF is added 5.00 g (28.1 mmol) ofN-bromosuccinimide. The resulting suspension is stirred at room températureovemight and concentrated. The solid is triturated with not water. then washed 25 with isopropanol to give 5.59 g 179.4%) of the product as a soiid. mp 266-270°C. EXAMPLE 74 6-Bromo-7-chloro-2-nietliylsulfanyl-pyrido[2,3-dJpyrimidine
By substituting the product of Example 73 in Example 6. 2.73 g (97.2%)of the product is obtained as a solid. 012161 -54- MS (APCI) M+l: Calcd 289.9; Found 289.8. EXAMPLE 75 6-Bromo-2-metliylsulfanyl-pyridof2.3-d]pyrimidin-7-ylamine
Bv substituting the product of Example 74 in Example 7. 2.09 g (82.9%) 5 of the product is obtained as a solid. MS (APCI) M+l; Calcd 271.0: Found 270.8. EXAMPLE 76 6-Bromo-2-methanesulfïnyl-pyrido[2,3~d]pyrimidin-7-ylamine
By substituting the product of Example 75 in Example 8. 1.81 g (81.9%) 10 of the product is obtained as a solid, mp 245°C (dec). MS (APCI) M+l : Calcd 287.0; Found 286.8. 1 EXAMPLE 77 4-[4-7-Aniino-6-bromo-pyrido[2,3-d]pyrimidin-2-ylatnino)-phenylJ-piperazine-1-carboxylic acid tert-butyl ester 15 By substituting the product of Example 76 in Example 13. 1.40 g (44.4%) of the product is obtained as a solid. MS (APCI) M+l: Calcd 500.1: Found 500.0. EXAMPLE 78 4-{4-[6-Bronio-7-(3-cyciohexyl-ureido)-pyrido[2.3-d)pyrimidin-2-ylamino]- 20 phenyl}-piperazine-l-carboxyHc acid tert-butyl ester
By substituting the product of Exampie 77 in Example 16. 0.1160 g (46.4%) of the product is obtained as a solid. MS (APCI) M+l : Calcd 625.2: Found 625.1. EXAMPLE 79 25 l-[6-Bromo-2-(4-piperazin-l-yl-plienylaniino)-pyrido[2,3-d]pyrunidiii-7-yl]-3- cydohexyl-urea
By substituting the product of Example 78 in Example 15. 0.0886 g(77.0%) of the product (compound 55) is obtained as a solid. mp 195°C (dec). 012161 -55- MS (APCI) M+l: Calcd 525.2: Found 525.1.
Anal. Calcd for C24H29BrjNgOj-1.64 Η2ΟΊ.83 HCl: C. 46.37: H. 5.53: N. 18.02: Cl. 10.44.
Found: C. 46.53: H. 5.34: N. 17.73: Cl. 10.15. EXAMPLE 80 4-{4-[6-Bromo-7-(3-tert-butyl-ureido)-pyrido[2,3-d]pyrimidiH-2-yIamino]-phenyl}-piperazine-l-carboxylic acid tert-butyl ester
Bv substituting the product of Example 77 in Example 10. 0.2571 g(42.9%) of the product is obtained as a solid. MS (APCI) M+l: Calcd 599.2: Found 599.2. EXAMPLE 81 l-l6-Bromo-2-(4-piperaziii-l-yI-phenylantino)-pyrido[2,3-d]pyritnidin-7-y[]-3- tert-butyl-urea
By substituting the product of Example 80 in Example 15, 0.0481 g of theproduct (compound 91) is obtained as a solid. MS (APCI) M-H : Calcd 499.2: Found 499.0. EXAMPLE 82 4-{4-[6-Bromo-7-(3-methyl-ureido)-pyrido[2,3-d]pyrimidin-2-ylaminoJ-phenyl}-piperazine-l-carboxylic acid tert-butyl ester
By substituting the product of Example 77 and methylamine inExample 32. 0.170 g (29.9%) of the product is obtained as a solid. MS (APCI) M+l: Calcd 557.2: Found 557.1. EXAMPLE 83 1 -l6-Bromo-2-(4-piperazin-l-yl-phenylamino)-pyrido[2,3-d]pyrimidin-7-yll-3-methyi-urea
By substituting the product of Example 82 in Example 15. 0.0963 g (69%)of the product (compound 93) is obtained as a solid. MS (APCI) M+l: Calcd 457.1: Found 457.1. -56- 012161
Anal. Calcd for CjçHojBrjNgOpS HC1-3 H2O: C. 36.76: H. 4.87: N. 18.05; Cl. 17.13: H2O. 8.71.
Found: C. 36.49: H. 4.35: N. 17.52; CI. 15.79: H2O. 8.12. EXAMPLE 84 5 4-[4-(7-Amino-6-bronw-pyrido[2,3-d]pyritnidin-2-ylantino)-phenyl}-cis-2,6-dimethyl-piperazine-l-carboxylic acid tert-butvi ester
By substituting the product of Example 76 in Example 27. 2.10 g (63.1%)of the product is obtained as a solid. MS (APCI) M-l: Calcd 528.2: Found 528.2. 10 EXAMPLE 85 4-{4-[6-Bromo-7-(3-tert-butyl-ureido)-pyrido[2,3-d]pyriinidin-2-yiamino}-phenyl}-cis-2,6-dimethyl-piperazine-l-carboxyUc acid tert-bufyl ester
By substituting the product of Example 84 in Example 10. 0.1725 g(72.6%) of the product is obtained as a solid. 15 MS (APCI) ΜΉ : Calcd 627.2: Found 627.2. EXAMPLE 86 J-{6-Bromo-2-l4-(cis-3,5-dimetbyl-piperazin-]-yl)-phenyiantino]-pyrido[2,3-dJpyrimidin- 7-yl}-3-tert-buty!-urea
By substituting the product of Example 85 in Example 15. 0.1593 g 20 (96.0%) of the product (compound 94) is obtained as a solid. mp 202°C (dec). MS (APCI) M+l: Calcd 527.2: Found 527.2.
Anal. Calcd for C24H3iBriNgOj-2.55 HCÎ-1.70 H2O: C. 44.28: H. 5.72: N. 17.21: Cl. 13.89.
Found: C. 44.28: H. 5.72: N. 17.09: Cl. 12.49. 012161 EXAMPLE 87 4-{4-[6-Bromo-7-(3-cyclohexyl-ureido)-pyrido[2.3-d]pyrimidin-2-ylcinùnoJ-phenyl}-cis-2,6-dimethyl-piperazine-l-carboxylic acid tert-butyl ester
Bv substituting the product of Example 84 in Example 16. 0.1750 g5 (70.7%) of the product is obtained as a solid. MS (APCI) M+l: Calcd 653.3; Found 653.3. EXAMPLE 88 l-{6-Bronio-2-[4-(cis-3,5-dimethyl-piperazin-l-yl)-phenylanüno]-pyrido[2,3- d]pyritnidin-7-yl}-3-cyclohexyi-urea 10 By substituting the product of Example 87 in Example 15. 0.1614 g (95.4%) of the product (compound 95) is obtained as a solid. mp 19S°C (dec). MS (APCI) M+l: Calcd 553.2: Found 553.2.
Anal. Calcd for Caé^NgOiBrj -2.76 HC1-2.02 H2O: C. 45.22: H. 5.81: N. 16.23: CL 14.17. 15 Found: C, 45.23: H. 5.82; X. 16.08: CL 13.53. EXAMPLE 89 N~-(4-Fiuoro-phenyl)-pyrido[2,3-d]pyrinùdine-2,7-diamhte
By substituting 4-fluoroaniline in Example 9. 1.1529 g (45.2% ) of the product is obtained as a solid. mp 245-248°C. 20 MS (APCI) M-l: Calcd 256.1: Found 255.9. EXAMPLE 90 l-[2-(4-Fluoro-phenylamino)-pyrido[2,3-d]pyrimidm-7-yll-3-(3-morpholin-4-yl- propyl)-urea
By substituting the product of Example 89 and 3-morphoIin-4-yl- 25 propylamine in Example 32. 0.1465 g (58.6%) of the product (compound 96) isobtained as a solid. mp 253-256°C. MS (APCI) M+l: Calcd 426.2: Found 426.1. 012161
Anal. Calcd for C21H24F1N7O2·' C. 59.28: H. 5.69: N. 23.04.
Found: C. 59.18: H. 5.66: N. 23.04. EXAMPLE 91 l-[2-(4-Fluoro-phenylamino)-pyrido{2,3-d]pyrimidin-7-yl]-3-(2-hydroxy-ethvl)- urea B y substituting the product of Example 89 and 2-hvdroxy-ethy lamine inExample 32. 0.0811 g (40.3%) of the product (compound 97) is obtained as asolid. mp 238-240°C. MS (APCI) M-l: Calcd 343.1: Found 343.1.
Anal. Calcd for CigHisFiN^CH: C. 56.14; H. 4.42: N. 24.55.
Found: C. 55.82: H. 4.52: N. 24.15. EXAMPLE 92 l-(2-Aimno-ethyl)-3-[2-(4-flitoro-phenylamiiio)-pyrido[2.3-d)pyrimidiii-7-yl}- urea
By substituting the product of Example 89 and ethylenediamine inExample 32. 0.1000 g (49.3%) of the product (compound 98) is obtained as asolid. mp217-220°C. MS (APCI) M-l : Calcd 342.1: Found 342.0.
Anal. Calcd for Ci6Hj6FjN7O]-0.2 FbO: C. 55.71; H. 4.79: N. 28.42.
Found: C. 55.72: H. 4.57: N. 28.07. EXAMPLE 93 l-(2-Diniethylamino-etliyl)-3-[2-(4-fluoro-pheitylamino)-pyrido[2,3- djpyrimidin- 7-yl]-ureci
By substituting the product of Example 89 and 2-dimethylamino-ethylamine in Example 32. 0.0778 g (35.8%) of the product (compound 99) isobtained as a solid. mp 251-255°C. PCT/IB* . 012161 MS (APCI) M+l: Calcd 370.2; Found 370.0.
Anal. Calcd forCig^oFlNyO]: C. 58.53: H. 5.46; N. 26.54.
Found: C. 58.39: H. 5.51: N. 26.26. 5 EXAMPLE 94 3,3-Dimethyl-l-(4-nitro-phenyl)-piperazine
By substituting 2.2-dimethyl-piperazine in Example 24,29.43 g (88.4%)of the produci is obtained as a solid. MS (APCI) M+l: Calcd 236: Found 236. 10 EXAMPLE 95 2-2-Dimethyl-4-(4-nitro-phenyl)-piperazine-l-carboxylic acid tert-butyl ester
By substituting the product of Example 94 in Example 11. 38 g (93%) ofthe product is obtained as a solid. MS (APCI) M+l: Calcd 336: Found 336. 15 EXAMPLE 96 4-(4-Amino-phenyl)-2,2-dimethyl-piperazine-l-carboxylic acid tert-butyl ester
By substituting the product of Example 95 in Example 12. 27 g (78%) ofthe product is obtained as a solid. MS (APCI) M+l: Calcd 306: Found 306. 20 EXAMPLE 97 4-[4-(7-Amino-6-fluoro-pyrido[2,3-d]pyrimidin-2-ylamino)-phenyl]-2,2-dimethyl-piperazine-l-carboxylic acid tert-butyl ester
By substituting the product of Example 96 in Example 50. 0.4346 g(59.0%) of the product is obtained as a solid. MS (APCI) M+l: Calcd 468.2: Found 468.3. 25 012161 W' -60- EXAMPLE 98 4-{4-[7-(3-Cyclohexyl-ureido)-6-fluoro-pyrido[2,3-d]pyrimidin-2-ylamiuo]-phenyl}-2,2-dimethyl-piperazine-l-carboxylic acid tert-butyl ester
By substituting the product of Example 97 in Example 16. 0.170 g (31.2%)5 of the product is obtained as a solid. MS (APCI) M+l: Calcd 593.2; Found 593.4. EXAMPLE 99 l-Cyclohexyl-3-{2-[4-(3,3-dimethyl-piperazin-I-yl)-phenylamino]-6-fluoro- pyrido[2,3-djpyritmditt-7-yl}-urea 10 By substituting the product of Example 98 in Example 15. 0.040 g of the product (compound 100) is obtained as a solid. MS (APCI) MH: Calcd 493.3; Found 493.2.
I EXAMPLE 100 4-[4-(7-Amino-6-fluoro-pyrido[2,3-d]pyrimidin-2-ylamino)-phenyl]-piperazine- 15 1-carboxylic acid tert-butyl ester
By substituting the product of Example 12 in Example 50. 0.2017 g (29.7%) of the product is obtained as a solid. MS (APCI) M+l: Calcd 440.2; Found 440.2. EXAMPLE 101 20 4-{4-[7-(3-Cyclohexyl-ureido)-6-fluoro-pyrido[2.3-dJpyrimidin-2-ylaminoJ-phenyl}-piperazine-l-carboxylic acid tert-butyl ester
By substituting the product of Example 100 in Example 16. 0.2036 g(78.6%) of the product is obtained as a solid. MS (APCI) MH: Calcd 565.3; Found 565.3. 25 EXAMPLE 102 ]-CycloliexyI-3-[6-fluoro-2-(4-piperazin-l-yl-plieiiylamiiio)-pyrido[2'3-djpyrimidin-7-ylj-urea
By substituting the product of Example 101 in Example 15. 0.1084 g(96.0%) of the product (compound 11) is obtained as a solid. 012161 -61- MS (APCI) M+l: Calcd 465.2: Found 465.2.
Anal. Calcd for C24H29FiNgO] -2.75 HC1-3.5 H2O: C. 45.91: H. 5.10; N. 17.85: Cl. 15.53: FbO, 10.04.
Found: C. 46.20: H. 5.86: N. 17.45: Cl. 15.22: H2O. 8.97. 5 EXAMPLE 103 4-{4-[7-(3-tert-Butyl-ureido)-6-fluoro-pyrido[2,3-d}pyrimidÎH-2-ylamino}-phenyi}-cis-2,6-dinteihyl piperazine-I-carboxylic acid tert-butyl ester
By substituting the product of Example 50 in Example 10. 0.070 g (17.9%)of the product is obtained as a solid. 10 MS (APCI) M+l: Calcd 567.3: Found 567.3. EXAMPLE 104 l-tert-Butyl-3-{2-l4-(cis-3J>-dimethyl-piperazin-l-yl)-phenylamino]-6-fluoro- pyrido[2.3~d]pyrimidin-7-yl}-urea
By substituting the product of Exampie 103 in Exampie 15. 0.0585 g of15 the product (compound 102) is obtained as a solid. MS (APCI) M+l : Calcd 467.3: Found 467.3. EXAMPLE 105 l-[4-(4-Nitro-phenyl)-piperazinyl}-ethanone
To a solution of 5.0 g (24.1 mmol) of l-(4-nitro-phenyl)-piperazine in 20 100 mL of dichloromethane was added 5.04 mL (28.9 mmol) of diisopropyl- ethylamine. The solution is cooled in an ice-bath. treated with 1.89 mL(26.5 mmol) of acetyl chloride. and stirred at room température ovemight. Thereaction is washed successively with water. 0.5 M HCl. saturated sodiumhydrogen carbonate, and brine. and dried over magnésium sulfate, and 25 concentrated to give 5.91 g (98.5%) of the product as a solid. MS (APCI) M-l: Calcd 250.1: Found 250.0. 012161 V. . -62- EXAMPLE 106 l-[4-(4-Amino-phenyl)-piperazin-l-yI]-ethanone
By substituting the product of Example 105 in Exampie 12. 4.35 g (84.1%)of the product is obtained as a soiid. 5 MS (APCI) M+l : Calcd 220.1 : Found 220.1. EXAMPLE 107 l-{4-[4-(7-Amino-pyrido[2,3-d]pyrimidin-2-ylamino)-phenyt]-piperazin-I-yl}- ethanone
By substituting the product. of Example 106 in Example 9. 0.1829 g10 (50.1 %) of the product is obtained as a soiid. MS (APCI) M+l: Calcd 364.2: Found 364.2.
Anal. Calcd for C]9H2iN7Oi-1.0H2O: C, 59.46; H. 6.11: N. 25.55.
Found: C, 59.51: H. 6.03; N. 25.28. 15 EXAMPLE 108 l-{2-[4-(4-AcetyI-piperazin-I-yI)-phenylamino]-pyrido[2,3-d]pyrimidin---yl}-3- (3-morpholin-4-yl-propyl)-urea
By substituting the product of Example 107 and 3-morpholin-4-vl-propylamine in Example 32. 0.0338 g (22.6%) of the product (compound 103) is 20 obtained as a soiid. mp 222-225°C (dec). MS (APCI) M+l: Calcd 534.3; Found 534.2.
Anal. Calcd for C27H35N9O3O.5 FbO: C. 59.76; H. 6.69; N. 23.25.
Found: C. 59.74; H. 6.53; N. 23.35. 25 EXAMPLE 109 6-Chloro-2-methylsuifanyl-8H-pyrido[2.3-d]pyrimidin-7-one
By substituting N-chlorosuccinimide in Example 74. 0.3700 g (3 1.4%) ofthe product is obtained as a soiid. mp 264-266°C (dec). MS (APCI) M+l: Calcd 228.0: Found 227.9. 012161 FC. -63- EXAMPLE 110 6/7-Dichioro-2-methylsulfanyl-pyrido[2,3-d]pyrimidine
By substituting the product of Example 109 in Example 6. 0.6534 g (86.5%) of the product is obtained as a soîid. 5 MS (APCI) M+l : Calcd 246.0; Found 245.8. EXAMPLE 111 6-Chloro-2-methylsulfanyl-pyrido[2,3-d]pyrimidin-7-ylamine
By substituting the product of Example 110 in Example 7. 0.38 g (63%) of the product is obtained as a solid. 10 MS (APCI) M+l : Calcd 227.0; Found 226.9. EXAMPLE 112 6-Chloro-2-methanesuIfinyI-pyrido[2,3-dJpyrimidin-7-ylamine^
By substituting the product of Example 111 in Example 8. 0.2328 g (57.1%) of the product is obtained as a solid. mp 260-262°C. 15 MS (APCI) M+l: Calcd 243.0; Found 242.9. EXAMPLE 113 4-[4-(7-Αηΰηο-6-αΙιΙο™-ρ\'ΓΪάο[2,3-ά]ρνήηήάιη-2-\Ίαιηΐηο)-ρΙιεηνΙ]-ϋΪ5-2,6-dintethyl-piperazine-î-carboxylic acid tert-butyl ester
By substituting the product of Example 312 in Example 27. 0.22 g (49%) 20 of the product is obtained as a solid. MS (APCI) M+l: Calcd 484.2: Found 484.2. EXAMPLE 114 4-{4-[7-(3-tert-Butyl-ureido)-6-chloro-pyrido{2,3-d]pyrimidin-2-ylamino}-phenyl}-cis-2,6-dimethyl-piperaziite-l-carboxylic acid tert-butyl ester 25 By substituting the product of Exampie 113 in Example 10. 0.0995 g (39.2%) of the product is obtained as a solid. 012161 τ -64- EXAMPLE 115 l~tert~Butyl-3-{6-chloro-2-[4~(cis-3,5-dimethyl-piperazin-l-yl)-phenylaniino}- pyrido[2,3-d]pyrimidin-7-yl}-urea
Bv substituting the product of Example 114 in Example 15. 0.0995 g of theproduct (compound 104) is obtained as a soîid. mp 205°C (dec). MS (APCI) M-rl: Calcd 483.2: Found 483.2. EXAMPLE 116
Methyl-(2-methylsulfanyl-pyrido[2,3-d]pyrimidin-7-yl)-ainine
By substituting methvlamine in Example 7. 1.46 g (30.0%) of the productis obtained as a solid. MS (APCI) M+l : Calcd 207.1 : Found 206.9. EXAMPLE 117 (2-Methanesulfinyl-pyrido[2,3-d]pyrimidin-7-yl)-methyl-amiHe
By substituting the product of Example 116 in Example 8. 1.31 g (83.4%)of the product is obtained as a solid. mp 185°C. MS (APCI) M+l: Calcd 223.1: Found 223.0. EXAMPLE 118 4-l4-(7-Methylamino-pyrido[2.3-d]pyrimidin-2-ylamino)-phenyl]-piperazine-l-carboxylic acid tert-buryl ester
By substituting the product of Example 117 in Example 13. 0.4934 g(62.9%) of the product is obtained as a solid. MS (APCI) M+l: Calcd 436.2; Found 436.2. EXAMPLE 119 4-{4-[7-(3-Cyclohexyl-l-methyl-ureido)-pyridof2,3-d]pyrimidin-2-yf]-piienyl}-piperazine-J-carboxylic acid tert-butyl ester
By substituting the product of Example 118 in Example 16. and usingacetonitrile as solvent and no base. 0.8535 g (78.8%) of the product is obtained asa solid. MS (APCI) M-l: Calcd 561.3: Found 561.3. 012161 -65- EXAMPLE 120 3-Cyclohexyl-l-methyl-l-[2-(4-piperazin-l-yl-phenylamino)-pyrido[2.3- djpyrimidin- 7-ylJ-urea
By substituting the product of Exampie 119 in Example 15. 0.2548 g(36.0%) of the product (compound 70) is obtained as a solid. mp 169-175°C. MS (APCI) M-î-3: Calcd 461.3: Found 461.2.
Anal. Calcd for C25H32NgO]O.25 H2O: C. 64.56: H. 7.04; N. 24.09.
Found: C. 64.57: H. 7.01: N. 23.98. EXAMPLE 121 3-Cyclohexyl-l-{2-[4-(cis-3,5-dimethyl-piperazin-l-yl)-phenylaminol-pyrido[2,3- dJpyrimidin-7-yl}-l-methyl-urea
Using the general procedure by which Example 120 is synthesized, 0.1366 g (95.6%) of the product (compound 106) is obtained as a solid,mp 170°C (dec). MS (APCI) M+l : Calcd 489.3: Found 489.3.
Anal. Calcd for C27H36N8Op3.32 ^0-2.69 HCl: C. 50.16: H. 7.07: N. 17.33: Ci. 14.75.
Found: C. 50.36: H. 6.98: N. 16.97: Cl. 15.07. EXAMPLE 122 3-CyclohexyI-l-ethyl-l-[2-(4-piperazin-l-y!-phenyIanüno)-pyrido[2,3-d]pyrimidin- 7-yl]-urea
Using the general procedure by which Example 120 is synthesized. 0.118g(94%) of the product (compound 107) is obtained as a solid. MS (APCI) M+l : Calcd 475.3: Found 475.3.
Anal. Calcd for C26H34NgO]-3.0 HC1-0.3 diethylether: C. 53.89: H. 6.65: N. 18.48.
Found: C. 53.75: H. 6.96: N. 18.57. 012161 -66- EXAMPLE 123 3-tert-Butyl-l-{2-[4-(cis-3,5-dimethyl~piperazin-l-yl)-phenylaminoJ-pyrido[2.3- d]pyrimidin-7-yl}-l-ethyl-urea
Using the general procedure by which Exampie 15 is synthesized. 0.022 c5 (56%) of the product (compound 108) is obtained as a soiid. MS (APCI) M+l: Calcd 477.3; Found 477.3. EXAMPLE 124 l-Metliyl-3-[5-methyl-2-(4-piperazin-l-yl-phenylamino)-pyrido[2,3-d]pyrimidin 7-yl]-urea 10 Using the general procedure by which Exampie 40 is synthesized. the product (compound 64) is obtained as a solid. mp 204-206°C (dec). MS (APCI) M-t-1: Calcd 393; Found 393. t EXAMPLE 125 l-Ethyl-3-[5-niethyI-2-(4-piperazin-]-yl-phenylainino)-pyrido[2,3-(ijpyrimidin- 15 7-yl]-urea
Using the general procedure by which Example 40 is synthesized. theproduct (compound 28) is obtained as a soiid. mp 220-222°C. MS (APCI) M+l: Calcd 407: Found 407. EXAMPLE 126 20 l-[5-Metliyl-2-(4-piperazin-l-yl-phenylamino)-pyrido[2,3-d]pyrimidin-7-y[]- 3-propyI-urea
Using the general procedure by which Example 40 is synthesized. theproduct (compound 111) is obtained as a soiid. mp 223-225°C. MS (APCI ) Μ* 1 : Calcd 421 ; Found 421. 012161 rci/ -67- EXAMPLE 127 N,N-Dimethyl-N'-[5-methyi-2~[[4-(l-piperazinyl)phenyl]-amino}-pyrido[2,3-djpyrimidin- 7-yl-sulfamide
Using the general procedure by which Example 40 is synthesized. butusing dimeihvl sulfamyl chloride rather than cyciohexylisocvanate. the product(compound 71) is obtained as a solid. mp 228-230°C (dec). MS (APCI) M+l: Calcd 443: Found 443. EXAMPLE 128 7-Amino-2-methylsulfanyl-pyrido[2,3-d]pyriinidine-6-carboxylic acid ethyl ester
To a solution of 4-amino-2-methanesulfanyI-pyrimidine-5-carboxaldehyde(Example 3) in 10 mL of tetrahvdrofuran is added 0.126 mL (1.18 mmol) of ethylcvanoacetate. The solution is cooled to -10°C, and treated with 2.36 mL(2.36 mmol) of titanium tetrachloride. To the solution is slowly hdded 0.52 mL(4.72 mmol) of N-methyl morphoiine. The reaction is warmed to roomtempérature over 2 hours. and partitioned between ethyl acetate and saturatedaqueous ammonium chloride. The organic laver is concentrated to give a solid,which is triturated with ether to give 0.30 g (96%) of the product as a solid. MS (APCI) M+l: Calcd 265.1: Found 264.9. EXAMPLE 129 7-Amino-2-chloro-pyrido[2,3-d]pyrimidine-6-carboxylic acid ethyl ester
To a suspension of the product of Example 128 in 50 mL of chloroform isslowly added sulfuryl chloride. followed by 2 drops of éthanol. The reaction isstirred at room température for 16 hours. poured into ether. and the solid collectedto give 0.50 g (98%) of the product. MS (APCI) M-*-l: Calcd 253.1: Found 253.1. EXAMPLE 130 7-Amino-2-[4-(4-tert-butoxycarbonyl-piperazin-l-yl)-phenylamino]-pyrido[2,3~dJpyrimidine-6-carboxylic acid ethyl ester A solution of the product of Example 12 and the product of Example 129in dioxane is heated under reflux for 1.5 hours. The reaction is poured into 012161 -68- hexane/ethyl acetate (1:1}. and the solid is collected. Flash chromatographv usinedichloromethane as eluant gave 0.08 g (16%) of the product as a solid. MS (APCI) M+l: Calcd 494.2: Found 494.1. EXAMPLE 131 2-[4-(4-tert-Butoxycarbonyl-piperazin-l-yI)-phenylamino]-7-(3-tert-butyl-ureido)-pyrido[2.3-d]pyrimidine-6-carboxylic acid ethyi ester
Bv substituting the product of Example 130 in Example 10. 0.05 g (48%)of the product as a solid is obtained. EXAMPLE 132 7-(3-tert-Butyl-ureido)-2-(4-piperazin-l-yl-phenylamino)-pyrido[2.3-dJpyrimidine-6-carboxylic acid etltyl ester
By substituting the product of Example 131 in Example ll 5. 0.036 g of theproduct (compound 113) as a solid is obtained. mp >300°C. EXAMPLE 133 l-[6-Fluoro-5-methyl-2-(4-piperazin-l-yl-phenylamino)-pyrido[2,3-d]pyrimidin- 7-yl]-3-isopropyl-urea
Using the general procedure by which Example 52 is synthesized. butusing l-(4-amino-2-methylsulfanyl-pyrimidin-5-yl)-ethanone (Example 35),4-(4-amino-phenvl)-piperazine-l-carboxyiic acid tert-butyl ester (Example 12). andisopropyl isocyanate as reagents, the product (compound 114) is obtained as asolid. mp 208°C (dec). MS (APCI) M+l : Calcd 439.2: Found 439.3. EXAMPLE 134 l-Cyclohexyl-3-{2-l4-(3.3-dinietliyI-piperazin-l-yl)-phenyiamino]-pyrido[2.3-djpyrimidin-7-ylJ-urea
Using the general procedure by which Example 17 is synthesized. butusing 4-(4-amino-phenyl)-2.2-dimethyl-piperazine-l-carboxylic acid tert-butylester (Example 96). 0.95 g (100%) of the product (compound 115) is obtained asa solid. 012161 -69- MS (APCI) M-H: Calcd 475.6: Found 475.3.
Anal. Calcd for C26H34N8O]-3 HCM H2O: C. 51.96: H. 6.37; N. 18.64: Cl. 17.69: FhO. 2.99.
Found: C. 52.00: H. 6.41: N. 18.53: Cl. 16.51; H2O. 3.06. EXAMPLE 135 6-Methyl-2-methyIsulfanyl~pyrido[2,3-d]pyrimidin-7-ylamine
To a suspension of 2.18 g (54 mmol) of 60% oil dispersed sodium hvdridein 300 mL of tetrahydrofuran. cooled to 10°C. is added 10.2 g (53.4 mmol) of(l-cyano-l-methyl-rneihyl)-phosphonic acid diethyl ester (Svnthesis. 1975:516).To the cooled suspension is added 4.30 g (25.4 mmol) of 4-amino-2-methanesulfanyl-pyrimidine-5-carboxaldehyde (Example 3). and the reaction isstirred at room température for 22 hours. The resulting solution is concentrated t and filtered to give a solid. which is washed with tetrahydrofuran. dissolved inIN citric acid, and re-precipitated by adjusting the pH to 8 with 50% sodiumhydroxide. The solid is collected by filtration to give 1.1 g (21%) of the product.mp 268-270°C. MS (APCI) M-H : Calcd 207.3: Found 207.0. EXAMPLE 136 l-Cyclohexyl-3-{2-l4-(cis-3,5-dimethyl-piperazin-l-yl)-phenylanünof-6-methyl- pyrido[2,3-d]pyrimidin-7-yl}-urea
Using the general procedure by which Example 31 is synthesized. butusing the product of Example 135 as the staning nrterial. 0.14 g (42%) of theproduct (compound 116) is obtained as a solid. MS (APCI) M+l : Calcd 589.6: Found 589.3.
Anal. Calcd for CgyH^gXgOi -2.5 HCI-1.5 HgO: C. 53.80: H. 6.73: N. 18.01: Cl. 14.11: HgO. 4.06.
Found: C. 53.44: H. 6.89: N. 18.46: Cl. 14.60: HgO. 4.48. 012161 -70- EXAMPLE 137 I-tert-Butyl-3-{2-[4-(cis-3,5-dimethyl-piperaziti-l-yl)-phenylamitio}-6-rnethyl- pyrido[2,3-d]pyrimidin-7-yl}-urea
Using the general procedure by which Example 29 is synthesized. butusing the product of Example 135 as the starting material. 0.26 g (89%) of theproduct (compound 117) is obtained as a solid. MS (APCI) M+l: Calcd 463.6: Found463.3.
Anal. Calcd for C25H36NgO]-2.4 HC1-1.75 FbO: C. 51.62: H. 6.91: N. 19.26; Cl. 14.63: H2O. 5.42.
Found: C. 51.23: H. 6.55: N. 18.92; Cl, 14.73: H2O. 5.10. EXAMPLE 138 l-lert-Butyi-3-[6-methyl-2-(4-piperazin-l-yl)-phenylamino)-pyrido[2,3- t djpyrimidin- 7-yl]-urea
Using the general procedure by which Example 15 is synthesized, butusing the product of Example 135 as the starting material. 1.02 g (100%) of theproduct (compound 118) is obtained as a solid. MS (APCI) M+I: Calcd 435.3: Found 435.3.
Anal. Calcd for C23H3oNgC>i-5 HCl· 1.75 H2O: C. 42.60: H. 5.98: N. 17.28: Cl. 27.34; H2O. 4.86.
Found: C. 42.03: H. 6.04; N. 16.81: CI. 22.95: H2O. 4.72. EX AMPLE 139 l-{2-[4-(cis-3,5^imethyl-piperazin-l-yl)-phenylamino}-6-methyl-pyrido[2.3- dJpyrimidin-7-yl}-3-isopropyl-urea
Using the general procedure by which Example 33 is synthesized. butusing the product of Example 135 as the starting material. along with 4-(4-amino-phenyl)-cis-2.6-dimethyl-piperazine-l-carboxylic acid tert-butvl ester andisopropvlamine as reagents. 0.130 g (100%) of the product (compound 119) isobtained as a solid. MS (APCI) M-H: Calcd 449.3; Found 449.3. 01 2161 -71-
Anal. Calcd for C24H32NgOi-3 HCl· 1.75 H2O: C. 48.90: H, 6.58; N. 19.01: Cl. 16.04; H2O. 5.35.
Found: C. 49.03; H, 6.63; N. 18.70: Cl. 16.03; H2O. 5.19. EXAMPLE 140 5 l-Cyclopropyl-3-{2-[4-(cis-3,5-dimethyl-piperazin-l-yl)-piienyÎanùno}-6-metiiyÎ-pyrido[2,3-d]pyrimidin-7-yl}-urea
Using the general procedure bv which Example 33 is svnthesized. butusing the product of Example 135 as the starting material. along with 4-(4-amino-phenyl)-cis-2.6-dimethyl-piperazine-l-carboxyiic acid tert-butvl ester and 10 cyclopropvlamine as reagents. 0.099 g ( 100%) of the product (compound 120) isobtained as a solid. MS (APCI) M+l: Calcd 447.3: Found 447.3.
Anal. Calcd for C24H3oNgOj : C. 49.83; H. 6.19: N. 19.37; Cl. 18.39; H2O, 3.89. 15 Found: C, 49.76: H. 6.23; N. 18.92; Cl, 15.66; H2O. 3.06. EXAMPLE 141 l-tert-Butyl-3-{2-{4-(cis-3,5-dimethyl-piperazin-I-yl)-plienylaminoj-6-ethyl- pyrido[2,3~dJpyrimidin-7-yl}-urea
Using the general procedure by which Example 137 is synthesized, but20 using ( 1-cyano-propyl)-phosphonic acid diethyl ester as starting material. 0.34 g (95%) of the product (compound 121) is obtained as a solid. MS (APCI) M+l: Calcd 477.3: Fourni 477.3.
Anal. Calcd for HC1-1 H2O: C. 53.26: H, 7.05: N. 19.11: Cl. 15.18: H2O. 3.07. 25 Found: C. 53.63: H. 7.31 : N. 18.46: Cl. 15.32: H2O. 3.48. EXAMPLE 142
The following compounds are prepared essentially according to theprocedures described in Examples 3-141 and shown in Schemes 1-4: 012161 -72- (a) 1 -tert-Butyl-3-[2-(3-chloro-4-piperazin-1 -yl-phenyiamino )-pyrido[2.3-d]pyrimidin-7-yl]-urea (compound 2); (b) 1 -tert-Butyl-3-[6-fluoro-2-(4-piperazin- î -vi-phenylamino)-pyrido[2.3-d]pyrimidin-7-yl]- urea (compound 3); 5 (c) l-{2-[4-(4-Acetyl-piperazin-l-yl)-3-chloro-phenylamino]- pyrido[2.3-d]pyrimidin-7-yl}-3-tert-butyI-urea (compound 6): (d) l-{2-[4-(4-Acetyl-piperazin-l-yl)-phenvlamino]-6-fluoro-pyrido[2.3-d]pyrimidin-7-yl}-3-tert-butyl-urea (compound 7): (e) 1 -{2-[4-(4-Acetvl-piperazin-l -yl)-phenylamino]-5-methvl-10 pyrido[2.3-d]pyrimidin-7-yl}-3-ten-butyl-urea (compound 8): (f) l-[2-(3-Chloro-4-piperazin-l -yl-phenylamino)-pyrido[2.3-d]pyrimidin-7-yl]-3-cyclohexyl-urea (compound 10): (g) l-{2-[4-(4-Acetyl-piperazin-l-yl)-phenylamino]-pyrido[2.3-d]pyrimidin-7-yl}-3-cyclohexyî-urea (compound 13)1. 15 (h) l-{2-[4-(4-Acetyl-piperazin-l-yl)-3-chloro-phenylamino]- pyrido[2.3-d]pyrimidin-7-yl}-3-cyclohexyl-urea (compound 14): (i) l-{2-[4-(4-Acetyl-piperazin-l-yl)-phenylamino]-6-fluoro-pyrido[2.3-d]pyrimidin-7-yl}-3-cyclohexyl-urea (compound 15): (j) 1 -(2-(4-(4-Acetyl-piperazin-1 -yl)-phenylamino}-5-methyl- 20 pyrido[2.3-d]pyrimidin-7-yl}-3-cyclohexyl-urea (compound 16): (k) l-(2-Hydroxy-ethyl)-3-[2-(4-piperazin-l-yl-phenylamino)-pyrido[2.3-d]pyriniidin-7-yl]-urea (compound 17): (l) 1 -[2-(3-Chloro-4-piperazin-1 -yl-phenvlamino)-pyrido[2.3-d]pyrimidin-7-yl]-3-(2-hydroxy-ethyD-urea (compound 18); 25 (m) l-[6-FIuoro-2-(4-piperazin-l-yl)-phenylamino)- pyrido[2.3-d]pyrimidin-7-yl]-3-(2-hydroxy-ethyI }-urea (compound 19); (η) 1 -(2-Hydroxy-ethyl)-3-[5-methyl-2-(4-piperazin-1 -yl- phenylamino)-pyrido[2.3-d]pyrimidin-7-yl]-urea (compound 20): (o) l-i2-[4-(4-Acetyl-piperazin-l-yl)-phenylamino]- 30 pyrido[2.3-d]pyrimidin-7-yl}-3-(2-hydroxy-ethyl)-urea (compound 21): (p) 1 - (2-[4-(4-Acetyl-piperazin-1 -yl )-3-chloro-phenylamino]-pyrido(2.3-d]pyrimidin-7-yl}-3-(2-hydroxy-ethyl)-urea (compound 22): 012161 -73- (q) 1 - {2-[4-(4-Aceivl-piperazin-l -yl)-phenylaminoJ-6-fluoro-pyrido[2.3-d]pyrimidin-7-yl}-3-(2-hydroxy-ethyl)-urea (compound 23): (r) 3 -{2-(4-(4-Aceiyl-pîperazin-1 -yl)-phenylamino]-5-methyl-pyrido[2.3-d]pyrimidin-7-yl}-3-(2-hydroxy-ethyl)-urea (compound 24): (s) 1 -Ethyl-3-[2-(4-piperazin-l -yl-phenylamino)-pyrido[2.3-djpyrimidin-7-yl]-urea (compound 25); (t) 1 -{2-(3-Chloro-4-piperazin-l -yl-phenylamino)-pyrido[2.3-d]pyrimidin-7-yI]-3-ethyl-urea (compound 26): (u) 1 -Ethyl-3-[6-fluoro-2-(4-piperazin-1 -yl-phenylamino)-pyrido[2.3-d}pyrimidin-7-yl]-urea (compound 27); (\') 1 - (2-[4-(4-Acetyl-piperazin-I-yi)-phenylamino]- pyrido[2.3-d]pyrimidin-7-yI}-3-ethyl-urea (compound 29): (w) l-{2-[4-(4-Acetyl-piperazin-l-yl)-3-chloro-phenyiamino]-pyrido[2.3-d]pyrimidin-7-yl}-3-ethyl-urea (compound 30): (x) 1 -{2-[4-(4-Acetyl-piperazin-l-yl)-phenylamino]-6-fiuoro-pyrido[2.3-d]pyrimidin-7-yl}-3-exhyl-urea (compound 31); (y) 1 - {2-[4-(4-AceTyl-piperazin-l -vl)-phenylamino]-5-methyl-pyrido[2.3-d]pyrimidin-7-yl}-3-eîhyl-urea (compound 32): (z) l-Îert-BuÎyl-3-[2-(pyridin-4-yîamino)-pyrido[2.3-d]pyrimidin-7-vI]-urea (compound 33): (aa) l-Cyclohexyl-3-[2-(pyridin-4-ylamino)-pyrido[2.3-d]pyrimidin-7-vl]-urea (compound 34): (bb) l-Ethyl-3-[2-(pyridin-4-yIamino)-pyrido[2.3-d]pyrimidin-7-yl]-urea (compound 35): (cc) 1 -(Hydroxy-ethyl)-3-[2-(pyridin-4-ylamino)-pyrido[2.3-d]pyrimidin-7-yl]-urea (compound 36); (dd) 1 -iert-But> l-3-[6-iluoro-2-(pyridin-4-ylamino)-pyrido[2.3-d]pyrimidin-7- yl]-urea (compound 37): (ee) 1 -Cyc!ohexyl-3-[6-fiuoro-2-(pyridin-4-ylamino)-pyrido[2.3-d]pyrimidin-7-yl]-urea (compound 38): (ff) 1 -Ethyl-3-[6-fluoro-2-(pyridin-4-yIamino)-pyrido[2.3-d]pyrimidin-7-yl]- urea (compound 39):
Wv 012161
PC -74- (gg) l-[6-Fluoro-2-(pyridin-4-yiamino)-pyrido[2.3-d]pyrimidin-7-yl]- 3-(2-hydroxv-ethyl)-urea (compound 40); (hh) 1 -tert-Butyl-3-[5-meihyl-2-(pyridin-4-ylamino)- pyrido[2.3-d]pyrimidin-7- yl]-urea (compound 41); 5 (ii) 1 -Cyclohexyl-3-[5-methyl-2-(pyridin-4-ylamino)- pyrido[2.3-d]pyrimidin-7-yi]-urea (compound 42); (jj) l-Ethyl-3-[5-methyl-2-(pyridin-4-ylamino)-pyrido[2,3-d]pvrimidin-7-yl]- urea (compound 43); (kk) l-(2-Hydroxy-ethyI)-3-[5-methyl-2-(pyridin-4-ylamino)-10 pyrido[2.3-d]pyrimidin-7-yl]-urea (compound 44); (11) 1 -Cyclohexvl-3-[6-methyl-2-(4-piperazin-1 -yl-phenvlamino)- pyrido [2.3-djpyrimidin-7-yl]-urea (compound 54); (mm) 1 -CyclohexyI-3-[6-cyano-2-(4-piperazin-1 -yl-phenylamino)-pvrido[2.3-d]pyrimidin-7-yI]-urea (compound 56); 15 (nn) 1 -Cyclohexyl-3-[6-chloro-2-(4-piperazin-1 -yl-phenylamino)- pyrido [2.3-d]pyrimidin-7-yl]-urea (compound 57): (oo) l-Cvclohexyl-3-[6-fluoro-5-methy 1-2-(4-piperazin-l -yl- phenylamino)-pyrido[2.3-d]pyrimidin-7-yl}-urea (compound 58); (pp) 1 -Cyciohexyl-3-[6-bromo-5-methyl-2-(4-piperazin-l -vl-20 phenylamino)-pyrido[2.3-d]pyrimidin-7-yi]-urea (compound 59): (qq) l-Cyclohexyl-3-[6-chioro-5-methyl-2-(4-piperazin-l-vl-phenylamino)-pyrido[2.3-d]pyrimidin-7-yl]-urea (compound 60): (rr) 1 -Isopropyl-3-[5-methyl-2-(4-piperazin-1 -yl-phenylamino)-pyrido[2.3-d]pyrimidin-7-yl]-urea (compound 61): 25 (ss) 1-[5-Methvl-2-(4-piperazin-l-yl-phenylamino)- pyrido(2.3-d]pyrimidin-7-yl]-urea (compound 63): (tt) 1 -(4-Hydroxy-cvclohexyl )-3-[2-(4-piperazin-1 -yl-phenyiamino)- pyrido [2.3-d]pyrimidin-7-vl]-urea (compound 66); (uu) l-(4-Amino-cyciohexyl)-3-[2-(4-piperazin-l-yl-phenylamino)-30 pvrido [2.3-d]pyrimidin-7-yl]-urea (compound 67): ( vv) 1 -(2-Dimethy lamino-ethyl)-3-[2-(4-piperazin-1 -y 1-phenv lamino)-pvrido [2.3-d]pyrimidin-7-yl]-urea (compound 68): 012161 -75- ( ww) 1 -(3-MorphoIino-4-yl-propy l)-3-[2-(4-piperazin-1 -y 1-phenylamino)-pyrido [2.3-d]pyrimidin-7-yI]-urea (compound 69): (xx) l-Cyclohexyl-3-[5-meîhyl-2-(4-piperazin-l-yl-phenylamino)-pvrido [2.3-d]pyrimidin-7-yl}-thiourea (compound 72): (}'?') N-[2-(4-Piperazin-l -yl-phenylamino)-pyrido[2.3-d]pyrimidin-7-vl]-acetamide (compound 73); (22) 4-[7-(3-Cyclohexyl-ureido)-pyrido[2.3-d]pyrimidin-2-ylamino]-ben2enesulfonamide (compound 74); (aaa) 1 -Cyclohexyl-3- {2-[4-( 1 -pipera2in- î -yl-methanoyl )-phenylamino] 10 pyrido[2,3-d]pyrimidin-7-yl)-urea (compound 75): (bbb) 1 -Cyclohexyl-3-[2-(4-fluoro-phenyiamino)- pyrido[2.3-d]pyrimidin-7-yl]-urea (compound 76); (ccc) l-(2-{4-[4-(2-Amino-4-methyl-pentanoyi)-pipera2in-l-vl]- phenylamino}-pyrido[2J-d]pyrimidin-7-yl)-3-cyclohexyl-urea (fcompound 77): 15 and (ddd) I-(2-{4-[4-(2-Amino-3-methyl-butanoyl)-piperazin-l-yI}-phenylamino}-pyrido[2.3-d]pyrimidin-7-yl)-3-cyclohexyl-urea (compound 78). EXAMPLE 143
Biological Assav 20 As noted above. the compounds of this invention are potent inhibitors of cdks. and accordingly. are useful in treating and preventine atherosclerosis andother cell proliférative disorders like cancer that are mediated bv such cdken2vmes. The compounds exhibit excellent inhibitory activity against a number ofcdk en2ymes. including cdkcdkl/cyclinB. cdk2/cyclinA. cdk2/cyclinE, and 25 cdk4/cvclinD. when evaluated in standard assays routinely utiii2ed by those skilled in the art to measure cdk inhibitory activities. Typical assays are carriedout as follows.
Cyclin Dépendent Kinase 4 (cdk4) Assay
En2yme assays for 1C5Q déterminations and kinetic évaluation are
30 performed in 96-well filter plates (Millipore MADVN6550). The total volume is0.1 mL containing a final concentration of 20 mM TRIS 01 21 61
PC -76- (tris[hydroxymethyl]aminomethane). ai pH 7.4. 50 mM NaCl. 1 mMdithiothreitol. 10 mM MgCb. 25 μΜ ATP containing 0.25 uCi of p-PjATP. 20 ne of cdk4. 1 pg of retinoblastoma. and appropriate dilutions of a compound of theprésent invention. Ail components except the ATP are added to the wells. and theplate is placed on a plate mixer for 2 minutes. The reaction is started bv adding[•’-PjATP. and the plate is incubated at 25°C for 15 minutes. The reaction isterminated by addition of 0.1 mL of 20% trichloroacetic acid (TC A). The plate iskept at 4°C for at least 1 hour to allow the substrate to precipitate. The wells arethen washed 5 times with 0.2 mL of 10% TCA and 32p incorporation isdetermined with a beta plate counter (Wallac Inc.. Gaithersburg. MD).
Cyclin Dépendent Kinase 1 and 2Assays (cdkl/cyclinB, cdk2/cyclinA,cdk2/cyclinE)
K
Enzyme assays for ICjq déterminations and kinetic évaluation are performed in a 96-well ftlter plate (Millipore MADVN6550) in a total volume of0.1 mL of 20 mM TRIS (tris[hydroxymethyl]aminomethane), at pH 7.4. 50 mMNaCl. 1 mM dithiothreitol. 10 mM MgCb. 12 mM ATP containing 0.25 pCi of [^-PJATP. 20 ng of enzyme (either cdkl/B. cdk2/A. or cdk2E). 1 pgretinoblastoma. and appropriate dilutions of the particular invention compound.
Ail components except the ATP are added to the wells. and the plate is placed on apiate mixer for 2 minutes. The reaction is begun by addition of p-P]ATP. and theplate is incubated at 25°C for 15 minutes. The reaction is terminated by additionof 0.1 mL of 20% TCA. The piate is kept at 4°C for at least 1 hour to allow thesubstrate to precipitate. The wells are then washed 5 times with 0.2 mL of 10%TCA and J-P incorporation determined with a beta plate counter fWallac Inc.,Gaithersburg. MD).
Cyclin Dépendent Kinase 5/p25 Proline-directed Protein Kinase Assay
Source of enzvme: recombinant baculovirus-infected insect cell sf9-expressed recombinant cdk5-p25 complex. 012161 -77-
Purpose: Το evaluate the ability of test agents to inhibit cdk5/p25phosphorylation of Histone Hl.
Method: Baculovirus-insect cell His-tagged cdk5/Glu-tagged p25 (orGST-p25) enzyme complex is diluted to a concentration of 50 ng/20 pL in 5 Enzyme Dilution Buffer (EDB - 50 mM Tris-HCl [pH 8.0]. 10 mM NaCl. 10 mMMgClo. and 1 mM DTT). A 20 pL sample of test agent (diluted in EDB) is then combined with 20 pL of the of the final cdk5/p25 enzyme préparation and allowedto stand for 5 minutes at room température. Twenty-five microiiters of substratesolution containing 115 pL/mL Histone Hl. 30 pM ATP (vanadate-free). and 10 30 pCi/mL γ-^^Ρ ATP (Amersham)'in EDB is then added to the test agenv'enzyme
préparation and shaken at 30°C for 45 minutes. A 50 pL sample of the finalpréparation is added to 100 mL of 150 mM phosphoric acid on ice for 30 minutesto facilitate précipitation. The precipitate is then filtered through a 96-wellphosphocellulose filter plate and subsequently rinsed 3 times with 75 mM 15 phosphoric acid. Each well then receives 20 pL of scintillation cocktail, and the plates are counted for beta émissions using a Trilux Counter (33p fiher protocol).Test samples are compared to Control (no test agent présent: as 0% inhibition) andBaseline level (no enzyme, no test agent: as 100% inhibition) beta émissions todétermine the percent inhibition of Histone Hl phosphorylation. 20 The results of the foregoing assays for représentative invention compounds are presented in Table 2 below. The invention compounds exhibit IC5Q valuesranging from 0.027 pM to >5 pM against cdkî/B. from 0.010 pM to >5 pMagainst cdk2/A. from 0.020 to >5 pM against cdk2/E. and from 0.004 to >5 pMagainst cdk4/D. The most potent compound overall is compound 9, which 25 exhibits IC50 values of 0.027 pM. 0.010 pM. 0.020 pM. 0.005 pM. againstcdkl/B. cdk2A. cdk2E. and cdk4D. respective!}·.
PCI 012167 TABLE 2. Inhibition of Cdks: IC50 (μΜ)
Compound Cdkl/B Cdk2/A Cdk2/E Cdk4D 1 0.219 0.060 0.130 0.006 4 >5 >5 >5 1.5 5 0.463 0.130 0.130 0.037 9 0.027 0.010 0.020 0.005 11 0.159 0.092 0.125 0.011 12 >5 ’ >5 >5 2.100 28 >5 >5 >5 >5 45 0.552 0.054 0.110 0.045 46 0.075 0.300 l >5 47 >5 >5 >5 >5 48 0.257 0.113 0.098 0.018 49 0.911 0.528 0.475 0.050 50 0.069 0.022 0.035 0.007 51 0.053 0.024 0.030 0.004 52 0.472 0.213 0.126 0.027 53 >5 >5 >5 >5 012161 -79- TABLE 2. Inhibition of Cdks: IC5Q (μΜ) (cont)
Compound Cdkl/B Cdk2/A Cdk2/E Cdk4D 55 >5 >5 >5 0.300 64 >5 >5 >5 >5 65 >5 >5 >5 >5 70 1.448 0.697 0.530 0.017 71 >5 >5 >5 >5 79 >5 1.066 >5 >5 80 0.461 0.092 0.230 0.460 81 2.610 1.560 3.250 0.500 t 83 0.399 0.305 0.315 0.055 84 >5 >5 >5 >5 85 >5 >5 >5 >5 86 >5 >5 >5 >5 87 >5 >5 >5 >5 88 0.418 0.043 0.055 0.025 89 >5 >5 >5 >5 91 >5 >5 >5 0.070 93 >5 >5 >5 >5 94 >5 0.101 95 >5 0.310 96 6.365 1.108 1.550 >5 97 0.862 0.278 0.345 >5 012161
V» P -80- TABLE 2. Inhibition of Cdks: IC50 (μΜ) (cont)
Compound Cdkl/B Cdk2/A Cdk2/E Cdk4D 98 0.442 0.157 0.140 1.050 99 1.810 1.012 0.410 >5 100 0.265 0.153 0.415 0.035 102 3.130 3.590 4.500 0.165 103 >5 >5 >5 >5 104 >5 >5 >5 0.185 106 0.350 0.440 107 1.728 1.950 1.650 0.019 108 2.425 2.035 3.050 0.067 111 >5 >5 >5 >5 113 >5 >5 >5 3.000 114 >5 >5 >5 >5 115 0.094 0.022 0.051 0.007 116 >5 >5 3.750 0.3 33 117 >5 >5 4.000 0.076 118 >5 >5 3.800 0.079 119 >5 >5 >5 1.600 120 >5 >5 >5 1.900 121 >5 >5 >5 0.092
The compounds of this invention also are inhibitors of the growth factorreceptor tyrosine kinase enzymes. FGFr and PDGFr. and of the nonreceptortyrosine kinase enzyme. c-Src. Several of the invention compounds hâve been 012161
’ PC -81- evaluated via standard assays that measure their abilitv to inhibit ryrosine kinaseenzymes. These assays are carried out as follows: PDGF and FGF Receptor Tyrosine Kinase Assays
Full-length cDNAs for the mouse PDGF-β and human FGF-1 (flg) 5 receptor tyrosine kinases are obtained from J. Escobedo and prepared as describedin J. Biol. Chem.. 1991;262:1482-1487. PCR primers are designed to amplify afragment of DNA that codes for the intracellular tyrosine kinase domain. Thefragment is inserted into a baculovirus vector. cotransfected with AcMNPV DNA.and the recombinant virus isolated. SF9 insect ceils are infected with the virus to 10 overexpress the protein. and the cell lysate is used for the assay. Assays areperformed in 96-well plates (100 pL/incubation/well). and conditions areoptimized to measure the incorporation of ^-P from γ^-Ρ-ΑΤΡ into a glutamate-tyrosine co-polvmer substrate. Briefly. to each well is added 82.5'pL of incubationbuffer containing 25 mM Hepes (pH 7.0). 150 mM NaCl. 0.1% Triton X-100. 15 0.2 mM PMSF. 0.2 mM Na3VO4.10 mM MnCH. and 750 pg/mL of Poiy (4:1) glutamate-tyrosine followed by 2.5 pL of inhibitor and 5 pL of enzyme lysate(7.5 pg/pL FGF-TK or 6.0 pg/pL PDGF-TK) to initiate the reaction. Following a10-minute incubation at 25°C. 10 mL of γ^-Ρ-ΑΤΡ (0.4 pCi plus 50 pM ATP) isadded to each well. and samples are incubated for an additional 10 minutes at 20 25°C. The reaction is terminated by the addition of 100 pL of 30% trichloroacetic acid (TCA) containing 20 mM sodium pyrophosphate and précipitation ofmaterial onto glass fïber mats (Wallac). Filters are washed 3 times with 15% TCAcontaining 100 mM sodium pyrophosphate, and the radioactivitv retained on thefilters counted in a Wallac 1250 Betaplate reader. Nonspecific activity is defined 25 as radioactivitv retained on the filters following incubation of samples with bufferalone (no enzyme). Spécifie enzvmatic activity (enzyme plus buffer) is defined astotal activity minus nonspecific activity. The % inhibition at 50 pM is determined.and for the more potent compounds the concentration of the compound thatinhibited spécifie activity by 50% (IC5q) is determined based on the inhibition 30 curve. -82- PC'j . 012161 C-Src kinase Assay C-Src kinase is purified from baculovirus infected insect cell lysâtes usinaan antipeptide monoclonal antibodv directed against the N-terminal amino acids(amino acids 2-17) of c-Src. The antibodv. covalently linked to 0.65 um latexbeads. is added to a suspension of insect cell Ivsis buffer comprised of 150 m.MNaCl. 50 mM Tris pH 7.5. 1 mM DTT. 1% NP-40. 2 mM EGTA. 1 mM sodiumvanadate. 1 mM PMSF. 1 pg/mL each of leupeptin. pepstatin. and aprotinin. Insectcell lysate containing c-Src protein is incubated with these beads for 3 to 4 hoursat 4°C with rotation. At the end of the lysate incubation, the beads are rinsed3 times in lvsis buffer. resuspendedin lysis buffer containing 10% glvcerol. andfrozen. These latex beads are thawed. rinsed 3 times in assay buffer (40 mM Tris.pH 7.5. 5 mM MgCl?), and suspended in the same buffer. In a Millipore 96-well plate with a 0.65 pm polyvinylidine membrane bottom are added the reaction1 components: 10 pL c-Src beads. 10 pL of 2.5 mg/mL poly GluTvr substrate. 5 pMATP containing 0.2 pCi labeled -^-P-ATP. 5 pL DMSO containing inhibitors or asa solvent control. and buffer to make the final volume 125 pL. The reaction isstarted at room température by addition of the ATP and quenched 10 minutes laterby the addition of 125 pL of 30% TCA. 0.1 M sodium pyrophosphate for5 minutes on ice. The plate is then filtered and the wells washed with two 250-mLaliquots of 15% TCA. 0.1 M pyrophosphate. The filters are then punched. countedin a Iiquid scintillation counter. and the data examined for inhibitory activity incomparison to a known inhibitor such as erbstatin. The method is also described inJ. Med. Chem., 1994:37:598-609.
The tyrosine kinase inhibitory activity for représentative inventioncompounds evaluated in the foregoing assays is given in Table 3. 61 -83- TABLE 3. Inhibition of Tyrosine Kinases: % Inhibition at 50 μ\1(IC50 [μΜ] in parenthesis if determined)
Compound PDGFr FGFr 1 94.4 (0.593) 93.7 9 89.8 il (0.131) (0.284) 45 21.9 67.4 46 17.5 19.5 47 10.5 55 (0.033) (0.151) 70 (0.536) (1.15) 80 18.6 117 (0.081) (0.061)
As noted above. the invention aiso provides pharmaceutical compositionscomprising an invention compound admixed with a carrier, diluent, or excipient.The foliowing examples illustrate typical compositions provided by this invention. EXAMPLE 144 5 A pharmaceutical compositions in the form of hard gelatin capsules for oral administration is prepared using the foliowing ingrédients:
Quantity (mg/capsuie) Active compound 250 Starch powder 200 Magnésium stéarate 10 Total 460
The above ingrédients are mixed and filled into hard gelatin capsules in460 mg quantities. A typical active ingrédient is l-isobutyl-3-[2-{(2-chloro-4-piperazin-l-vI)-phenylamino}-pyrido[2.3-d]pyrimidin-7-yl}-urea. Thecomposition is administered from 2 to 4 limes a day for treatment of postsurgicalrestenosis. 10 PO '
-84- EXAMPLE 144a Compositions for Oral Suspension 01216 Ingrédient Amount l-lsopropyl-3-[5-methyi-6-bromo-2-(3-ethyipyridin-4- 500 mg ylamino)-pyrido-[2.3-d]pyrimidin-7-yl]-urea Sorbitol solution (70% NF) 40 mL Sodium benzoate 150 mg Saccharin 10 mg Cherry' flavor 50 mg Distilled water q.s. ad 100 mL
The sorbitol solution is added to 40 mL of distilled waier. and thepvridopyrimidine is suspended therein. The saccharin. sodium benzoate. andflavoring are added and dissolved. The volume is adjusted to 100 mL withdistilled water. Each milliliter of syrup contains 5 mg of active ingrédient. EXAMPLE 144b
Tablets Each Containing 60 mg of Active Ingrédient
Active ingrédient 60 mg
Starch 45 mg
Microcrystalline cellulose 55 mg
Polyvinylpyrrolidone (as 10% solution in water) 4 mg
Sodium carboxymethyl starch 4.5 mg
Magnésium stéarate 0.5 mg
Talc 1.0 mg
Total 150mg
The active ingrédient, starch. and cellulose are passed through a No. 45mesh US sieve and mixed thoroughly. The solution of polyvinylpyrrolidone ismixed with the résultant powders and then passed through a No. 14 mesh USsieve. The granules are dried at 50°C to 60°C and passed through a No. 3 8 meshUS sieve. The sodium carboxymethyl starch. magnésium stéarate, and talc. 10 012161 -85- previouslv passed through a No. 60 mesh US sieve. are then added to the granuleswhich. after mixing. are compressed on a tablet machine to yield tablets eachweighing 150 mg. A tvpical active ingrédient utiiized in the above préparation is the5 compound of Example 40 (Compound 12). This composition is weil-suited to treatment of diabetic retinopathy. EXAMPLE 144c A parentéral composition suitable for administration by injection isprepared by dissolving 100 mg of compound 77 in 250 mL of 0.9% aqueous 10 sodium chloride solution and adjusting the pH of the solution to about 7.0. Thisformulation is weil-suited for the treatment of breast cancer. EXAMPLE 144d 1
Préparation for Suppositories A mixture of 500 mg of l-n-butyl-3-[2-(4-piperazin-l -vl-phenylamino)-15 pyrido[2.3-d]pyrimidin-7-yl]-urea and 1500 mg of theobroma oil are blended to uniformity at 60°C. The mixture is cooled to 24°C in tapered molds. Eachsuppository will weigh about 2 g and can be administered from 1 to 2 times eachday for treatment of viral infections such as herpes and ΗΙλ7. -86- EXAMPLE 144e Topicaî Préparation r» 01216 Ingrédient Amount (mg) l-Cyclohexyl-3-{[2-(4-morphoiin-l-vl-phenylamino)]-5.6- 20 difluoro-pyrido[2.3-d]pyrimidin-7-yl} -urea Propylene glvcol 100 White Petrolatum 500 Cetearyl alcohol 50 Glvceryl stéarate 100 PEG 100 stéarate 100 Ceteth-20 50 Monobasic sodium phosphate 80 Total 1000
The invention compound is blended to uniformity with the otheringrédients to make a thick suspension. The suspension is applied evenly to anadhesive backed polymeric film and eut into a 2-inch square. The patch is applied 5 to the skin of a patient suffering from psoriasis. EXAMPLE 144f
Slow Release Préparation
Five hundred milligrams of 7-acetamido-6-bromo-2-[4-(2-diethylaminoethoxy)-phenylamino]pyrido[2.3-d]pyrimidine hydrochloride was 10 placed in an osmotic pump tabiet and administered oraliy to a subject fortreatment and prévention of restenosis.
The invention and the manner and process of making and using it are nowdescribed in such full. clear. concise and exact terms as to enable an\ personskilled in the art to which it pertains to make and use the same. it is to be 15 understood that the foregoing describes preferred embodiments of the présent invention, and that modifications may be made therein without departing from the spirit or scope of the présent invention as set forth in the daims. To particularlv 01 2161 PC7 -87- point out and distinctly claim the subject marrer regarded as the invention, thefollowing daims condude this spécification.

Claims (15)

  1. 012161 Pt -88- CLAIMS What is claimed is:
  2. 3. A compound of the Formula III
    and pharmaceuticaliy acceptable salts. esters, amides. and prodrugsthereof. wherein: R- is hydrogen. or lower alkyl. lower alkenvl. or lower alkynyi. each of which isoptionally substituted with up to 5 groups independentlvselected from halogen. cyano. nitro. -R9. -\R9RlO. -OR9.-<CH2 )nCO2R9. -(CH2)nSO2R 11. -(CH2)nR I ’. -COR9-CONR9R1Ü. -SO3R9. -SO2NR9Rl0. -SO2R9. -SR9, 012161 -93- 10 15 20 -PO3R9R10. -POR9r10. -PO(NR9R]0b--NR9COR10.-NR9CO2R10. -NR9CONR9RJÛ. -NR9SO2R10. ora heterocvcle optionallv substituted with up to 3 groups independently seiected from -R9. -NR9R^. -OR9.-(CH2)nSO2RI 1 - -(CH2)nRl1. or -(CH^JnR1- optionallv substituted with up to 5 groups independently seiected from halogen. cvano. nitro. -R9.-NR9R10. -OR9. -fCH2)nCO2R9. -(CH2)nSO2R15.-(CHolnR1 J. -COR9. -CONR9R10. -SO3R9-SO2NR9Rî0. -SO2R9. -SR9. -PO3R9R30. -por9rîo,-PO(NR9R10)2, -NR9CORlO. -nr9co2r10.-NR9CONR9RÏ0 -NR9SO2R10? or a heterocvcle optionallv substituted with up to 3 groups independently seiected from -R9. -NR9R1 θ, -OR9.-fCH2)nSO2R11.-(CH2)nRIi: R 5 is halogen. cyano. nitro. -R9. -NR9R^. or -OR9: R6 is halogen. cyano. nitro. -R9. -NR9R’ θ. -OR9. -CO2R9. -COR9, -CONR9R10. -NR9COR10. -so2nr9rJ°. -SO2R9. -SO3R9 SR9. -PO3R9R10. -POR9Rî0. -PO<NR9R10)2. or lower alkenyl or lower alkynyî optionallv substituted with -R9: R9 and R^ are independently hydrogen. or lower alkyl. optionallv substituted with up to 3 groups seiected from the group consisting of halogen. amino. mono- ordialkylamino. hydroxy. lower alkoxy. phenyl or substitutedphenyl. or when taken together with the nitrogen to which thev areattached. R9 and R^ form a ring having from3-7 members. up to four of which ma\ be seiected from 25 -94- 012161 q . O. S. and NR^O. where R-θ is hvdroeen. lower alkyl, or -CO lower alkyl; R11 is a heteroaryl or a heterocyclic group: R1 - is a cycloalkyl. a heterocyclic. an arvl. or a heteroaryl group;n is 0. 1.2. or 3; andRl9 îs hydrogen. or lower alkyl. lower alkenyl. or lower alkynyl. each of which isoptionallv substituted with up to 5 groups independentlvselected from halogen. amino. mono- or dialkylamino.hydroxy. lower alkoxy. cyano. nitro, carboxy. carboxvalkyl.aminocarbonyl. mono- or dialkylaminocarbonyl. loweralkylcarbonyl. -SO3R9. -SO2NR9r10. -SO2R9. -SR9,-POsR9R10. -POR9R10. -PO(NR9Ri0)2. -NR9COR10,-NR9CO2R10. -NR9CONR9RlO? -NR9SO2R10. or aryl. heteroaryl. arylalkyl. heteroarylalkvi. cycloalkyl or cycloaîkyl-alkyl. where each aryl. heteroaryl or cycloalkylgroup is optionallv substituted with up to 5 groupsindependentlv selected from halogen. amino. mono- ordialkylamino. hydroxy. lower alkoxy. cyano. nitro. carboxy,carboxvalkyl. aminocarbonyl. mono- or dialkylaminocarbonyl. alkylcarbonyl. -SO3R9. -SO2R9R10. -so2nr9rio. -SO2R9. -SR9. -PO3R9R10,-POR9R10. -PO(NR9Rî0)2. -NR9COR10. -NR9CO2R10.-NR9CONR9RlO. -NR9SO2R’0. or a (CH2)n- carbocyclic group containing from 3-7 members, up to 2 of which members are heteroatoms selected from oxygenand nitrogen. wherein the carbocyclic group isunsubstituted or substituted with 1. 2. or 3 groupsindependentlv selected from the group consisting of 01 2161 -95- haiogen. hvdroxv. lower alkyl. trifluoromethyl. loweralkoxy, amino. mono- or dialkylamino. aiyl. heteroaryl.arvlalkyl. heteroarylalkyl, heteroarylsulfonyl,heteroarylsuîfonylalkyl. heterocvclylalkyl.heterocyclylsulfonvl. or heterocyclylsulfonylalkyi: and R.21 is hydrogen. lower alkyl. or lower alkyl substituted with phenyl orsubstituted phenyl.
  3. 4. A compound of the Formula IV
    and pharmaceutically acceptable salts. esters, amides. and prodrugsthereof. wherein: R? is haiogen. cyano. nitro. -R9. -NR^RlO. or -OR9; r6 is haiogen. cyano. nitro. -R9. -NR^rIO. -OR9. -CO2R9. -COR9. -CONR9R!0. -NR9COR10. -so2r9r10. -SO2R9, -SO3R9. SR9,-PO3R9R10, POR9Rî0. -PO(NR9RlO)2. or lower alkenvl or lower alkynvl optionally substituted with -R9: R9 and RIO are independentiy hydrogen. or lower alkyl optionally substituted with up to 3 groups selected from the group consistingof haiogen. amino. mono- or dialkylamino. hvdroxv. lower alkoxy,phenyl or substituted phenyl. or when taken together with the nitrogen to which thev areattached. R9 and R1 θ form a ring having from 012161 -96- 3-7 members. up to four of which may be selected fromO C , O, S, and NR-θ, where R-θ is hvdrosen. lower alkyl. or -CO lower alkyl: Ri 1 is heteroaryl or a heierocyclic group: Rl 6. Rl 7. and R^ are independently selected from halogen. cyano. nitro. -R9, -NR9r!0 -OR9 -(CH2)nCO2R9. -(CHn^SOnR1 L-(CH2)nRlk -COR9 -CONR9RlO -SO3R9 -SO2NR9Rl0.-SO2R9. -SR9. -PO3R9Rl°. -POR9Ri0. -PO(NR9r10)2.-NR9COR1û. -NR9CO2R10. -NR9CONR9R]0.-NR9SO2Rl0, ora heterocycle optionally substituted with up to 3 groups independently selected from -R9, -NR9Rl9, -OR9,-NR9COR1 °. -COR1 θ -(CH2)nSO2R11. -(CH2)nR 11 ; Rl9 is hydrogen. or lower alkyl. lower alkenyl. or lower alkvnyl. each of which isoptionally substituted with up to 5 groups independentlyselected from halogen. amino. mono- or dialkvlamino.hydroxy. lower alkoxy. cyano. nitro. carboxy. carboxyalkyl.aminocarbonyl. mono- or dialkylaminocarbonvl. loweralkylcarbonyl. -SO3R9. -SO2NR9Rlû. -SO2R9. -SR9.-PO3R9R^. -POR9R1û. -PO(NR9RlO)2. -NR9COR10, -NR9CO2R10. -NR9CONR9R10. -NR9S02rIO. oraryl. heieroaryl. arylalkyl. heteroarylalkvl. cvcloalkyl or cycloalkyl-alkyl. where each aryl. heteroaryl or cycloaîkylgroup is optionally substituted with up to 5 groupsindependently selected from halogen. amino. mono- ordialkvlamino. hydroxy. lower alkoxy. cyano. nitro. carboxy,carboxyalkyl. aminocarbonyl. mono- ordialkylaminocarbonvl. alkylcarbonyl. -SO3R9. -97- 012161 '1 -SO2R9R10. -SOnNR^^. -SO2R9· -SR9. -PO3R9R’0.-POR9RlO -PO(NR9RlO)2- -NR9CORw. -nr9co2r10.-NR9CONR9R1û, -NR9SO2R10. or a (CH2)n* carbocyclic group containing from 3-7 members. up to2 of which members are heteroatoms selected from oxvgenand nitrogen. wherein the carbocyclic group isunsubsîituted or substituted with 1. 2. or 3 groupsindependently selected from the group consisting ofhalogen. hvdroxy. lower alkyi. trifluoromethyl. loweralkoxy. amino. mono- or dialkylamino. aryl. heteroaryl.arylalkyL heteroarylalkyl. heteroaryisulfonyl. heteroarvlsulfonylalkyl. heterocyclylalkyi.heterocyciylsuifonyi. or heterocyclylsulfonylalkyi: and R-1 is hvdrogen. lower alkyl. or lower alkyi substituted withphenyi or substituted phenyl. A compound of the Formula V
    and the pharmaceuticaily acceptable salts. esters, amides. and prodrugsthereof: wherein: R 5 is halogen. cyano. nitro. -R9. -NR9R^. or -OR9: R6 is halogen. cyano. nitro. -R9. -NR9Rl°. -OR9. -CO2R9. -COR9.-CONR9R10. -NR9CORl°. -SO2NR9R10. -SO2R9. -SO3R9.SR9. -PO3R9Rl 0. -POR9R1 θ. -POfNR9R 1 θ )2- or 012161 -98- lower alkenvl or lower alkynyl optionally’ substituted with -R9:r16, r!7_ 2^ r18 are independently selected from hydrogen. halogen. cvano. niiro. -R9, -NR9RlO -OR9 -(CH2)nCO2R9-(CH2)nSO2R11, -(CH2)nRn· -COR9, -CONR9R19. -SO3R9.-so2nr9r10. -SO2R9- -SR9 -po3r9rio -por9rjo.-PO(NR9R10)2? -NR9COR10. -NR9CO2R10. -NR9CONR9RlO-NR9SO2Riû. or a heierocvcle optionally substituted with up to 3 groups independently selected from -R9. -NR9r10. -OR9.-NR^ORiO -CORlO -(CH2)nSO2R11. R9 and Ri® are independently hydrogen. or lower alkyl optionally substituted with up to 3 groups selected from the group consistingof halogen. amino. mono- or dialkvlamino. hvdroxy. lower alkoxy.phenyl or substituted phenyl. or w'nen taken together with the nitrogen to which thev areattached. R9 and Ri θ form a ring having from 3-7 members. up to four of which may be selected from O ç . O. S. and NR-θ. where R-θ is hydrogen. lower alkyl. or -CO lower alkyl: R11 is heteroaryl or a heterocyclic group: R19 is hydrogen. or lower alkyl. lower alkenyl. or lower alkynyl. each of which isoptionally substituted with up to 5 groups independentlyselected from halogen. amino. mono- or dialkvlamino.hvdroxy. lower alkoxy. cvano. nitro. carboxy. carboxyalkvl.aminocarbonvl. mono- or dialkvlaminocarbonyl. loweralkylcarbonvl. -SO3R9. -SO2NR9r10. -SO2R9. -SR9. 012161 -99- -PO3R9R10. -POR9RlO -PO(NR9R10)2. -NR9COR1Û-NR9CO2R10. -NR9CONR9r1°. -NR9SO2Ri0. or aryl. heteroaryl, arylalkyl. heteroarylalkyk cycloalkvl or cvcloalkyl-alkyl. where each aryl. heteroaryl or cycloalkvl 5 group is optionally substiruted with up to 5 groups independently selected from halogen. amino. mono- ordialkylamino, hydroxy. lower alkoxv. cyano. nitro. carboxy.carboxyalkyi. aminocarbonyl. mono- or dialkylaminocarbonvl. alkvlcarbonyl. -SO3R9.
  4. 10 -SO2R9R10. -SO2NR9Rl0. -SO2R9. -SR9. -PO3R9Rlû -POR9r1°. -PO(NR9Ri0)2. -NR9COR10. -NR9CO2Rl0-NR9CONR9RlO -NR9SO2Rl0 or a (CH2)n- carbocvclic group containing from 3-7 members. up to2 of which members are heteroatoms selected from oxvgen 15 and nitrogen. wherein the carbocvclic group is unsubstituted or substituted with 1. 2. or 3 groupsindependently selected from the group consisting ofhalogen. hydroxy. lower alkyl. trifluoromethyl. loweralkoxv. amino. mono- or dialkylamino. aryl. heteroaryl. 20 arylalkyl. heteroarylalkyk heteroarylsulfonyl. heteroarylsulfonylalkyl. heterocyclylalkykheterocyclylsulfonyk or heterocyclylsulfonvlalkyl: and R-1 is hvdrogen. lower alkyl. or lower alkyl substituted with phenvl orsubstituted phenvl. -100 012161 A compound of Formula VI
    10 and the pharmaceutically acceptable salts. esters, amides. and prodrugs thereof: wherein: R? is halogen. cyano. nitro. -R9, -NR^RlO. or -OR9: R6 is halogen. cyano. nitro. -R9. -NR9R10. -OR9. -CO2R9. -COR9, -CONR9R10. -NR9COR19. -SO2NR9Rl0. -SO2R9. -SO3R9,SR9. -P03R9R10. -POR9R10. -PO(NR9RlO)2. or lower alkenyl or lower alkynvl optionally substituted with -R9;Rl? and R1^ are independentiy selected from halogen. cyano. nitro. -R9. -NR9RÏû -OR9. -fCH2)nCO2R9. -(CH2)nSO2RH. -fCHolnR11·-COR9. -CONR9RlO. -SO3R9. -SO2NR9R10. -SO2R9. -SR9,-PO3R9Rl0. -POR9r10. -PO(NR9R10)2. -NR9COR10.-NR9CO2R10. -NR9CONR9R10. -NR9SO2R10. ora heterocycle optionally substituted with up to 3 groups independentiy selected from -R9. -NR9R19. -OR9.-NR9COR’°. -COR1 θ. -(CHolnSOoR11. -(CH2)nR’1 ; R— and R---1 independentiy are hydrogen or alkyl /. A compound selected from: 012161 -101- 1 -tert-Butyl-3-[2-(4-piperazin-1 -yi-phenylamino)-pyrido[2.3-d]pynmidin-7-yl]-urea; 1 -tert-Butyl-3-[2-(3-chloro-4-piperazin-1 -vl-phenv lamino)-pyrido(2.3-d]pyrimidin-7-yl]-urea: l-tert-Butyl-3-[6-fluoro-2-(4-piperazin-l-yl-phenylamino)- pyrido(2.3-d]pyrimidin-7-yl]-urea: 1 -tert-Butyl-3-[5-methyl-2-(4-piperazin-1 -yl-phenylamino)-pyrido[2.3-d]pyrimidin-7-vl]-urea: 1-(2-(4-(4-Acetyl-piperazin-1-yl)-phenylamino]-pyrido(2.3-d]pyrimidin-7-yI}-3-tert-butyl-urea: l-{2-(4-(4-Aceryl-piperazin-l-yl)-3-chloro-phenylamino]- pyrido[2.3-djpyrimidin-7-yI}-3-tert-buÎyi-urea: 1 - (2-(4-(4-Acetyl -piperazin-1 -y 1 )-pheny lamino]-6-fluoro-pyrido(2.3-d]pyrimidin-7-yl}-3-tert-butyl-urea; 1 -(2-(4-(4- Acetyl-piperazin-1 -yl)-phenylamino]-5-methyl-pyrido[2.3-d]pyrimidin-7-yl}-3-tert-butyl-urea; l-Cyclohexyl-3-(2-(4-piperazin-î-yl-phenvlamino)-pvrido (2.3 -djpyrimidin- 7-y l]-urea: 1 -(2-(3-Chloro-4-piperazin-l -yl-phenylamino)-pyrido(2.3-d]pyrimidin-7-vl]-3-cyclohexyl-urea: l-Cyclohexyl-3-(6-fluoro-2-(4-piperazin-l-yl-phenylamino)- pyrido(2.3-d]pyrimidin-7-yl]-urea: ]-Cyclohexyl-3-[5-methyl-2-(4-piperazin-l-yI-phenylamino)- pyrido(2.3-d]pyrimidin-7-yl]-urea: 3 - (2-[4-(4-Acetyl-piperazin-l -yl )-phenylamino]-pyrido[2.3-d]pyrimidin-7-yl}-3-cyclohexyl-urea: 1 - {2-(4-(4- Acetyl-piperazin-1 -yl )-3-chloro-pheny iamino]-pyrido(2.3-d]pyrimidin-7-yl *, -3-cyclohexyi-urea; l-î2-{4-(4-Acetyl-piperazin-]-yl)-phenylamino]-6-fluoro- pyrido[2.3-d]pyrimidin-7-yl}-3-cyclohexyl-urea: 1 - {2-(4-(4-Acetyl-piperazin-1 -yl)-phenvlamino]-5-methyl-pyrido(2.3-d]pyrimîdin-7-yl}-3-cyclohexyl-urea: 012161 -102- 1 -(2-Hydroxy-ethyl)-3-[2-(4-piperazin-1 -yl-phenvlamino)-pyrido[2.3-d]pyriraidin-7-yl]-urea: 1 -[2-(3-Chioro-4-piperazin-1 -yl-phenyiamino)-pyrido[2.3-d]pyrimidin-7-yI]-3-(2-hydroxy-ethyl)-urea: 1 -[6-Fluoro-2-(4-piperazin-3 -yl)-phenylamino)-pyrido[2.3-d]pyrimidin-7-yl]-3-(2-hydroxy-ethyl}-urea: 1 -(2-Hvdroxy-ethyl)-3-[5-methyl-2-(4-piperazin-1 -yl-phenylamino)-pyrido[2.3-d]pyrimidin-7-yl]-urea: 1 - (2-(4-(4-Acetyl-piperazin-1 -yl)-phenylamino]-pyrido[2.3-d]pyrimidin-7-yl}-3-(2-hydroxy-ethyl)-urea: l-{2-[4-(4-Acetyl-piperazin-l-yl)-3-chloro-phenylamino]- pyrido[2.3-d]pyrimidin-7-yl}-3-(2-hydroxy-ethyl)-urea; 1 - {2-(4-(4- Acetyi-piperazin-1 -yl)-phenylamino]-6-fluoro-pyrido[2.3-d]pyrimidin-7-yl}-3-(2-hydroxy-ethyl)-urea;i l-{2-[4-(4-Acetyl-piperazin-l-yl)-phenylamino]-5-methyl- pyrido[2.3-d]pyrimidin-7-yl}-3-(2-hydroxy-ethyl)-urea:
  5. 11. Use of a compound according to Claim 1 in the manufacture of amédicament for controlling proliférative disorders selected from the group 10 consisting of cancer, psoriasis, vascular smooth muscle prolifération associated with a disorder selected from the group consisting ofatherosclerosis, postsurgical vascular stenosis, and restenosis in mammals.
  6. 12. Use of a compound according to Claim 1 in the manufacture of a médicament for inhibiting a cdk enzyme. 15
  7. 13. Use of Claim 12 wherein said cdk is cdkl.
  8. 14. Use of Claim 12 wherein said cdk is cdk2.
  9. 15. Use of Claim 12 wherein said cdk is cdk4.
  10. 16. Use of a compound according to Claim 1 in the manufacture of a 20 médicament for inhibiting a growth factor-mediated tyrosine kinase.
  11. 17. Use of Claim 16 wherein said growth factor-mediated tyrosine kinase isplatelet derived growth factor (PDGF). 012161 -109-
  12. 18. Use of Claim 16 wherein said growth factor-mediated tyrosine kinase isfibroblast growth factor (FGF).
  13. 19. Use of a compound according to Claim 1 in the manufacture of amédicament for treating a subject suffering from diseases caused byvascular smooth muscle cell prolifération.
  14. 20. Use of a compound according to Claim 1 in the manufacture of amédicament for treating a subject suffering from cancer.
  15. 21. Use of a compound according to Claim 1 in the manufacture of amédicament for treating a subject suffering from a cancer related frombreast cancer, large cell carcinoma, pancréas, colon, melanoma, lung andleukemia.
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