cpd 1 has now been found to effectively inhibit tyrosine kinases , other than ret / ptc1 , that play a central role in tumor onset , progression and spreading to distant organs . specifically , cpd 1 has been shown to completely inhibit the autophosphorylation of ret / men2a ( mutations c634r and c634w ), ret / men2b ( mutm918t ), met , pdgf - r , fgf - r1 , fgf - r3 and kit ( c - kit and mutδ559 ) tyrosine kinases , to reduce their expression ( down - regulation ) and to revert the phenotype of cells thereby transformed . object of the invention is therefore the use of the compound ( e )- 1 , 3 - dihydro - 5 , 6 - dimethoxy - 3 -[( 4 - hydroxyphenyl ) methylene ]- 2h - indol - 2 - one , or of its salts with pharmaceutically acceptable bases , for the treatment of tumors involving at least one of met , pdgf - r , fgf - r1 , fgf - r3 and kit tyrosine kinases , or involving at least one oncoprotein of the ret family , including ret receptors carrying men2 - associated mutations , in the initial stages of cell transformation or in the following stages of tumor proliferation and dissemination . the invention also concerns the use of any stereoisomer or tautomeric form of cpd . 1 . inhibition of deregulated , constitutively active , ret receptors is useful in the treatment of sporadic medullary thyroid carcinomas ( mtc ) and men2 - associated diseases including mtc , pheochromocytoma , parathyroid hyperplasia , and enteric ganglioneuromas . met inhibition is useful to antagonize the invasive / metastatic phenotype of tumors of epithelial origin . with respect to met - activating alterations , cpd 1 may also have a specific indication in the therapy of kidney tumors . kit inhibition is useful in the treatment of gastrointestinal stromal tumors , small cell lung carcinomas , seminomas and hematological malignancies such as mastocytosis and acute myelogenous leukemia . the uncontrolled activation of pdgf - r and its involvement in autocrine loops support the therapeutic use of cpd 1 in tumors unresponsive to conventional therapies , such as glioma and dermatofibrosarcoma protuberans . in addition , pdgf - r is involved in tumor angiogenesis and vascular development thus supporting the use of cpd1 for the control of neoangiogenesis in solid tumors . cpd 1 can be used for the treatment of melanomas and gliomas expressing high levels of fgf - r1 and of the respective bfgf ligand eventually involved in autocrine loops . since this receptor has an important role in angiogenic processes , its inhibition by means of cpd 1 is useful for the control of tumor vascularization . fgf - r3 inhibition is useful in the treatment of multiple myeloma , bladder and cervix carcinomas . for use in therapy , rpi - 1 and its salts can be formulated with pharmaceutically acceptable vehicles and excipients . the phenol function of cpd 1 can be salified by treatment with suitable organic or inorganic bases . the pharmaceutical compositions can be administered by the oral , parenteral , sublingual or transdermic routes , preferably in the form of tablets , capsules , granules , powders , syrups , solutions , suspensions , suppositories , controlled release forms . the compositions can be prepared with conventional techniques , using ingredients known in the art . the quantity of active principle can be varied depending on the severity of the disease , age of the patient , type and route of administration , but in general an amount of 0 . 1 to 1000 mg / kg , preferably from 5 to 300 mg / kg , more preferably from 20 to 200 mg / kg , in single or multiple doses one or more times a day , is used . ret oncoproteins carrying aminoacid substitutions which cause constitutive tyrosine kinase activity are involved in sporadic mtc and in the inherited type 2 multiple endocrine neoplasia syndromes ( men2a , men2b and familial mtc ), all characterized by the occurrence of mtc ( jhiang s . m . et al . oncogene 19 , 5590 , 2000 ). whereas in sporadic mtc ret mutations are somatic , in men2 patients ret mutations are present at the germline level . these mutations cause constitutive activation of the receptor without modifying its localization at the cell membrane . the ret oncoproteins used in this study involve cys634 ( indicated as ret / men2a c634r and ret / men2a c634w ) or met918 ( indicated as ret / men2b m918t ), and represent the most frequently expressed ret oncoproteins in men2a and men2b , respectively . the inhibitory effect of cpd 1 has been demonstrated in murine cells transfected with the ret / men2a ( c634r ) gene ( nih3t3 men2a ( c634r ) cells ) and in the human medullary thyroid carcinoma cell lines tt and mz - crc - 1 , respectively characterized by the expression of ret / men2a ( c634w ) and ret / men2b ( m918t ) ( fig1 ). a reduction of the oncoprotein tyrosine - phosphorylation and expression is observed in these cell lines ( fig1 c and 2a ). the inhibition of ret / men2a and ret / men2b receptor autophosphorylation by cpd 1 is associated with an antiproliferative effect ( fig1 b and 2b ). nih3t3 men2a ( c634r ) transfectants reverted their transformed phenotype following exposure to cpd1 ( fig1 a ). a significant dose - dependent antitumor activity has been observed in nude mice xenografted with the tt tumor : after oral administration of a daily dose of 50 - 100 mg / kg ( twice a day ), cpd 1 treatment reached 80 % tumor weight inhibition ( twi ) without inducing toxicity ( fig3 ). the demonstrated pharmacological and biochemical efficacy of cpd 1 in controlling the proliferation of mtc cells is particularly important considering the aggressiveness of such tumors and the inefficacy of conventional therapies . met , the hepatocyte growth factor receptor , is a protein tyrosine kinase involved in the invasive process characteristic of tumor progression and metastatic growth ( maulik g . et al . cytok . growth factor rev . 13 , 41 , 2000 ). alterations such as mutations , overexpression or the involvement in autocrine loops are the cause of uncontrolled constitutive activation of the kinase . the uncontrolled kinase activity of met is involved in the invasiveness of many tumors of epithelial origin . in papillary thyroid carcinomas met is frequently overexpressed . the results illustrated in fig4 show a dose - dependent inhibition of met autophosphorylation in the papillary thyroid carcinoma cell line tpc - 1 treated with cpd 1 . met protein levels are also reduced in treated cells . other receptor tyrosine kinases , such as pdgf - r ( rosenkranz s . and kazlauskas a . growth factors 16 , 201 , 1999 ) and fgf - r1 ( powers c . j . et al . endocr . rel . cancer 7 , 165 , 2000 ), which are involved either in autocrine loops or in neoangiogenic processes , have an important role in cancer growth . a deregulated activation of these receptors is observed in tumors unresponsive to conventional therapies , such as gliomas and melanomas . the results illustrated in fig5 and 6 show a dose - dependent inhibition by cpd 1 of receptor autophosphorylation induced by autocrine stimulation ( fig5 a ) or by exogenous ligand ( fig5 b and 6a ). these effects are associated with a reduced receptor expression . concentrations of cpd 1 higher than 15 μm cause full inhibition of pdgf - r phosphorylation . activating mutations of the fgf - r3 receptor tyrosine kinase such as chromosomal traslocations or point mutations produce deregulated , constitutively active , fgf - r3 receptors which have been involved in multiple mieloma and in bladder and cervix carcinomas ( powers c . j . et al . endocr . rel . cancer 7 , 165 , 2000 ). the ability of cpd 1 to down regulate both tyrosine autophosphorylation and expression of fgf - r3 ( muty373c ) exogenously expressed in nih3t3 transfectants is illustrated in fig6 b . kit tyrosine kinase is constitutively activated as a consequence of mutations or of its involvement in autocrine loops in different tumors such as small cell lung cancers , gastro - intestinal stromal tumors , seminomas and leukemias ( heinrich m . c . et al . j . clin . oncol . 20 , 1692 , 2002 ). as shown in fig7 , cpd 1 inhibits the constitutive autophosphorylation and expression of the kit ( δ559 ) mutant exogenously expressed in nih3t3 cells ( fig7 b ). such inhibition is associated with reversion of the transformed morphologic phenotype of the transfected cells ( fig7 a ). in addition , fig7 c shows the dose - dependent inhibition by cpd 1 of c - kit activated through autocrine loop in the small cell lung carcinoma cell line n592 . as used herein , the term “ tumor ” is intended to encompass but is not limited to the abnormal cell proliferation of malignant or non - malignant cells of various tissues and / or organs such as muscle , bone or connective tissue , the skin , brain , lungs , sex organs , the lymphatic or renal systems , mammary or blood cells , liver , the digestive system , pancreas and thyroid or adrenal glands . the abnormal cell proliferation can include but is not limited to tumors of the ovary , breast , brain , prostate , colon , liver , lung , ovary , uterus , cervix , pancreas , gastrointestinal tract , head , neck , nasopharynx , skin , bladder , stomach , kidney or testicles , kaposi &# 39 ; s sarcoma , cholangiocarcinoma , choriocarcinoma , neuroblastoma , wilms &# 39 ; tumor , hodgkin &# 39 ; s disease , melanoma , multiple myeloma , chronic lymphocytic leukemia and acute or chronic granulocytic lymphoma . the compounds according to the present invention can be administered alone or in combination with other anti - tumor or anti - cancer agents including but not limited to : adriamycin , daunomycin , methotrexate , vincristin , 6 - mercaptopurine , cytosine arabinoside , cyclophosphamide , 5 - fu , hexamethylmelamine , carboplatin , cisplatin , idarubycin , paclitaxel , docetaxel , topotecan , irinotecam , gemcitabine , l_pam , bcnu and vp - 16 . the compounds according to the present invention can also be included in a kit for the treatment of tumors . the kit can include additional anti - cancer or anti - tumor agents . fig1 . effects of cpd 1 on nih3t3 men2a ( c634r ) transfectants expressing exogenous ret / men2a ( c634r ). a ) reversion of the transformed morphologic phenotype of nih3t3 men2a ( c634r ) cells . cells were exposed to 6 μm cpd1 for 24 h and then photographed under a phase - contrast microscope ( original magnification × 100 ). b ) antiproliferative effect . nih3t3 men2a ( c634r ) cells and the parental nih3t3 cells were treated with increasing concentrations of the drug for 72 h and then counted with a coulter counter . dose - response curves , from which the reported ic 50 values were calculated , showed the higher sensitivity to the drug of the ret oncoprotein - positive cell line . c ) inhibition of ret / men2a ( c634r ) autophosphorylation and expression . cells were treated with solvent (−) or 10 μm cpd1 (+), for the indicated times . whole cell lysates were prepared and subjected to sds - page and western blotting with anti - ptyr antibody . after stripping , the filter was reblotted with anti - ret antibody . arrows indicate the partial and fully glycosilated forms of the ret / men2a receptor . following 2 h of exposure to the drug , the receptor appeared partially dephosphorylated . at longer times , a complete tyrosine dephosphorylation was associated with reduced expression of the receptor fig2 . effects of cpd1 on human mtc cell lines harboring men2 - associated ret mutants . a ) inhibition of ret autophosphorylation . tt and mz - crc - 1 cells , expressing respectively ret / men2a ( c634w ) and ret / men2b ( m918t ), were treated with the indicated concentrations of cpd1 , for 24 h . control cells ( c ) received the solvent . whole cell extracts were processed for western blotting and probed with anti - ptyr and anti - ret antibodies . as evidenced in anti - ptyr blots , cell treatment with the compound induced a dose - dependent inhibition of tyrosine phosphorylation of ret receptors in both cell lines . a reduced expression of receptor concentrations was observed in cells treated with the highest concentrations of the drug . b ) antiproliferative effect . tt and mz - crc - 1 cells were treated with increasing concentrations of cpd 1 for 7 days and then counted with a coulter counter . dose - response curves documented the ability of the drug to interfere with the proliferation potential of the two mtc cell lines . fig3 . antitumor activity of cpd 1 in nude mice harboring tt medullary thyroid carcinoma xenografts . drug treatment started 25 days after s . c . inoculum of the tumor cells . cpd 1 was delivered per os at 50 or 100 mg / kg , twice in a day ( 2qd ), for 10 consecutive days ( indicated by arrows ). control mice received the vehicle . the treatment induced a significant dose - dependent inhibition of tumor growth . twi were 60 % ( p & lt ; 0 . 005 ) and 80 % ( p & lt ; 0 . 0005 ) for the 50 mg / kg and 100 mg / kg doses , respectively . fig4 . inhibition of met autophosphorylation and expression in human papillary thyroid carcinoma cells ( tpc - 1 ) treated with cpd1 . a ): tpc - 1 cells were exposed to solvent (−) or the indicated concentrations of cpd 1 for 72 h . equal amounts of protein were used for immunoprecipitation ( ip ) with anti - met antibody or for the preparation of whole cell lysates ( wcl ). immunoprecipitated proteins and wcls were separated by sds - page , transferred to nitrocellulose membranes , and subjected to western blotting with anti - ptyr or anti - met antibodies . b ): cells were treated as in a . cell extracts were immunoprecipitated with anti - ptyr antibody and probed with anti - met antibody . c ): cells were serum - starved for 24 h and exposed to solvent or cpd1 ( 60 μm ) during the last 18 h . then they were left untreated (−) or stimulated with 20 ng / ml hgf (+), for 10 min . cell lysates were immunoprecipitated with anti - met and probed with anti - ptyr or anti - met antibody . drug treatment abolished the constitutive or hgf - induced met tyrosine phosphorylation and induced a reduction of met expression . fig5 . inhibition of pdgf - r autophosphorylation and expression in whole cells by cpd 1 . a ): 2n5a cells ( nih3t3 transformed by the col1a1 / pdgfb rearrangement generating autocrine stimulation of pdgf - r ) were treated with cpd 1 at the indicated concentrations , for 72 h . cell extracts were immunoprecipitated with anti - pdgf - r and subjected to western blotting with anti - ptyr or anti - pdgf - r antibodies . b ): swiss 3t3 cells were serum starved for 24 h and then treated with cpd1 at the indicated concentrations , for 18 h . after stimulation with 1 nm pdgf for 5 min , whole cell lysates were prepared and subjected to western blotting with anti - ptyr or anti - pdgf - r . protein loading by anti - actin blot is shown . in both cell systems , a dose - dependent drug - induced inhibition of receptor tyrosine phosphorylation and expression was documented . fig6 . inhibition of autophosphorylation and expression of fgf - r1 and fgf - r3 receptors in whole cells by cpd1 . a ): swiss3t3 cells were serum starved for 24 h and then treated with cpd1 at the indicated concentrations , for 18 h . after stimulation with 100 ng / ml fgf for 5 min , whole cell lysates were prepared and subjected to western blotting with anti - ptyr or anti - fgf - r1 . protein loading by anti - actin blot is shown . b ): nih3t3 transformed by the fgf - r3 mutant y373c were treated with the indicated concentrations , for 72 h . cell extracts were immunoprecipitated with anti - fgf - r3 and then subjected to western blotting with anti p - tyr or anti - fgf - r3 . in both cell systems , a dose - dependent inhibition by cpd 1 of the receptor tyrosine phosphorylation and expression was documented . fig7 . effects of cpd1 on cell lines expressing constitutively activated forms of kit . a ): reversion of the transformed morphologic phenotype of nih3t3 transfectants expressing exogenous mutated kit ( δ559 ). cells were treated with 20 μm cpd1 and photographed 24 h later under a phase - contrast microscope ( original magnification × 100 ). b ): inhibition of kit ( δ559 ) autophosphorylation and expression in nih3t3 transfectants . cells were treated with cpd 1 at the indicated concentrations , for 72 h . cell lysates were immunoprecipitated with anti - kit antibody and subjected to western blotting with anti - p - tyr or anti - kit antibodies . c ): inhibition of expression and autocrine loop - activated autophosphorylation of c - kit in the sclc cell line n592 . cells were treated with cpd 1 at the indicated concentrations , for 24 h . whole cell lysates were subjected to western blotting with anti - kit antibody or with an antibody specifically recognizing the tyrosine phosphorylated / activated kit ( p - kit ). in both cell systems , a dose - dependent drug - induced inhibition of receptor tyrosine phosphorylation and expression was documented . the following human medullary thyroid carcinoma ( mtc ) cell lines were used in this study . the tt cell line derived from a men2a - associated mtc characterized by expression of the ret oncogene carrying the mutation c634w . the mz - crc - 1 cell line derived from men2b - associated mtc characterized by expression of the ret oncogene carrying the mutation m918t . tt cells were cultivated in ham &# 39 ; s f12 medium ( biowhittaker , verviers , belgium ) supplemented with 15 % fetal calf serum ( fcs ) ( life technologies , inc ., gaithersburg , md .) whereas mz - crc - 1 cells were grown in dulbecco &# 39 ; s modified eagle &# 39 ; s medium ( dmem ) ( biowhittaker ) supplemented with 10 % fcs . the human papillary thyroid carcinoma cell line tpc - 1 which was used as a model of met overexpression , was routinely cultivated in dmem with 10 % fcs . n592 cells , derived from a human sclc , were characterized by kit activation through autocrine stimulation by the scf ligand . n592 cells were cultured in rpmi 1640 supplemented with 10 % fcs . the mouse swiss3t3 and nih3t3 fibroblasts were cultured in dmem with 10 % calf serum ( colorado serum company , denver , colo .). in addition , nih3t3 cells transfected with different oncogenes were used . nih3t3 men2a transfectants express the short isoform of the ret - men2a ( c634r ) oncogene ; nih3t3 kitδ559 cells express kit carrying the activating mutation δ559 found in gists ; nih3t3 fgfr3 ( y373c ) express the fgf - r3 gene carrying the activating mutation y373c found in a human multiple myeloma cell line . 2n5a cells are nih3t3 cells transformed by the col1a1 / pdgfb rearrangement generating autocrine stimulation of pdgf - r . all nih3t3 transfectants were maintained in dmem plus 5 % calf serum in a 10 % co 2 atmosphere . cells were trypsinized after 3 days ( nih3t3 and nih3t3 men2a cells ) or 7 days ( mtc cells ) of treatment with cpd1 and counted by a coulter counter ( coulter electronics , luton u . k .). the concentrations able to inhibit cell proliferation by 50 % ( ic 50 ) were calculated from the dose - response curves . each experiment was performed in duplicate the following polyclonal antibodies were used : anti - ret recognizing a cooh - terminal sequence ( aa 1000 - 1014 ) common to the two ret isoforms ( borrello m . g ., et al ., mol . cell biol . 16 , 2151 , 1996 ); anti - ckit , anti - met and anti - fgf - r3 from santa cruz biotechnology ( santa cruz , calif . ); anti - actin from sigma ( st . louis , mo . ); anti - pdgf - r α / β from upstate biotechnology ( lake placid , n . y . ); anti phospho - ckit ( tyr 719 ) from cell signaling technology ( beverly , mass .). the mouse monoclonal anti - phosphotyrosine ( anti - ptyr ) 4g10 and anti - fgf - r1 antibodies were from upstate biotechnology . for whole - cell extract preparation , cells were lysed in sodium dodecyl sulfate ( sds ) sample buffer ( 62 . 5 mm tris - hcl ( ph 6 . 8 ), 2 % sds ) with 1 mm pmsf , 10 μg / ml pepstatin , 12 . 5 μg / ml leupeptin , 100 kiu aprotinin , 1 mm sodium orthovanadate , 1 mm sodium molybdate . protein concentration was determined in appropriately diluted aliquots by the bicinchionic acid ( bca ) method ( pierce , rockford , ill . ), then samples were adjusted to a final concentration of 10 % glycerol , 5 % β - mercaptoethanol , 0 . 001 % bromophenol blue . for immunoprecipitation experiments , cells were treated with cpd1 or solvent for the indicated times . cell monolayers were rinsed twice with cold phosphate - buffered saline plus 0 . 1 mm sodium orthovanadate and then left for 20 min on ice in lysis buffer ( 50 mm hepes ph7 . 6 , 150 mm nacl , 10 % glycerol , 1 % triton x - 100 , 1 . 5 mm mgcl 2 , 1 mm egta , 100 mm naf , 10 mm sodium pyrophosphate , 10 μg / ml antipain , 20 μg / ml chymostatin , 10 μg / ml e64 , 1 mg / ml pefabloc sc ). cells were then collected , aspirated through a 22 - gauge needle and centrifuged at 10000 g , for 20 min , at 4 ° c . protein concentration was determined by the bca reagent ( pierce ). cell extracts were incubated with protein a - agarose and the indicated antibody for 2 hr at 4 ° c . under rotation . after washing 3 times with 20 mm hepes ph 7 . 6 , 150 mm nacl , 10 % glycerol , 0 . 1 % triton x - 100 , the immunoprecipitates were eluted with complete sample buffer . normalized immunoprecipitates ( 0 . 5 - 4 mg of cell extracts ) or whole - cell lysates ( 30 - 60 μg ) were resolved on sds - page and transferred to nitrocellulose filters . membranes were incubated with primary antibodies at 4 ° c ., overnight . immunoreactive bands were revealed by horseradish peroxidase - coniugated anti - mouse or anti - rabbit antibodies using enhanced chemiluminescence detection systems from amersham biosciences ( little chalfont , united kingdom ) or pierce . all experiments were carried out using female athymic nude cd - 1 mice , 8 - 11 - weeks old ( charles river , calco , italy ). mice were maintained in laminar flow rooms with constant temperature and humidity . experimental protocols were approved by the ethic committee for animal experimentation of the istituto nazionale per lo studio e la cura dei tumori ( milan , italy ), according to the united kingdom coordinating committee on cancer research guidelines ( workman p . et al ., british journal of cancer , 77 , 1 , 1998 ). the mtc cells tt ( 1 . 6 × 10 7 cells ) were s . c . inoculated in mice as a cell suspension from in vitro cell culture . each control or drug - treated group included 8 - 10 tumors . tumor cells were injected on day 0 , and tumor growth was followed by biweekly measurements of tumor diameters with a vernier caliper . tumor weight ( tw ) was calculated according to the formula : tw ( mg )= tumor volume ( mm 3 )= d 2 × d / 2 , where d and d are the shortest and the longest diameter , respectively . drug treatment started when tumors were just measurable ( mean tw about 50 mg ), 25 days after the inoculum of tumor cells . cpd1 was dissolved in 5 % ethanol , 5 % cremophor el , 90 % saline ( 0 . 9 % nacl ) and delivered per os twice in a day ( 2qd ) by a daily schedule for 10 days . control mice received the solvent solution . drug efficacy was assessed as percentage tw inhibition ( twi % in drug - treated versus control mice expressed as : twi %= 100 −( mean tw treated / mean tw control × 100 ). for statistical analysis , tw in control and treated mice were compared on the day of twi % evaluation , by student &# 39 ; s t test ( two - tailed ). p values less than 0 . 05 were considered statistically significant . 45 g ( 0 . 23 moles , 1 eq .) of 3 , 4 - dimethoxyphenylacetic acid were dissolved in 100 ml ( 2 . 2 volumes ) glacial acetic acid at 28 ° c .- 35 ° c ., under n 2 atmosphere and mechanic stirring . the solution was cooled to 15 °- 20 ° c . and added with a mixture of fuming nitric acid ( 98 %, 33 ml ) in glacial acetic acid ( 25 ml ) over a period of 45 ′. once the addition was completed , precipitation of a red solid was observed . the suspension was poured in ice water ( 600 ml ) and kept under stirring for 2 h . the solid was filtrated , washed with water and dried at 60 ° c . for 8 h . 44 g of the end product were obtained . tlc ( sio 2 ; ethyl acetate 10 / acoh 0 . 5 ) rf acid = 0 . 6 ; rf product = 0 . 5 1h - nmr , ( dmso ): 3 . 9 ppm ( s , 6h ); 4 . 0 ppm ( s ., 2h ); 7 . 12 ppm ( s ., 1h ); 7 . 7 ppm ( s ., 1h ) 9 . 2 g ( 38 . 14 mmoles , 1 eq .) of 3 , 4 - dimethoxy - 2 - nitro - phenylacetic acid were suspended in glacial acetic acid ( 92 ml , 10 volumes ) at 25 ° c . under n 2 atmosphere and mechanic stirring . the suspension was added with fe ° powder , 325 mesh , 97 % ( 12 . 0 g , 214 . 86 mmoles , 5 . 6 eq .) in two equal portions . the first portion was added at room temperature ; then the mixture was refluxed and 30 min later the second fe ° portion was added . 30 ′ later the reaction was complete , tlc ( sio 2 ; chcl 3 9 / meoh 1 ), rf nitro = 0 . 65 , rf indolinone = 0 . 71 . the grey suspension was cooled to room temperature , the acetic acid was evaporated under low pressure to a crude solid , which was suspended in chloroform ( 200 ml ). the salts were filtrated off and the organic phase was washed with a nacl saturated solution ( 100 ml ), dried over na 2 so 4 and evaporated to dryness . the solid was suspended in ethyl ether ( 35 ml ) for 30 ′, filtrated and dried at 50 ° c . for 2 h . 6 . 7 g of a beige solid were obtained . tlc ( sio 2 ; ethyl acetate 10 / acoh 0 . 5 ) rf acid = 0 . 6 ; rf product = 0 . 5 1h - nmr , ( dmso ): 3 . 4 ppm ( s , 2h ); 3 . 69 ppm ( s ., 3h ); 3 . 72 ppm ( s ., 3h ); 6 . 49 ppm ( s ., 1h ); 6 . 92 ppm ( s ., 1h ); 10 . 15 ( s ., 1h ). 6 . 7 g ( 36 . 9 mmoles , 1 eq .) of 1 , 3 - dihydro - 5 , 6 - dimethoxy -( 1h )- indol - 2 - one were dissolved in anhydrous dmso ( 50 ml ) at room temperature . the solution was added with 4 - hydroxybenzaldehyde ( 5 . 41 g , 44 . 3 mmoles , 1 . 2 eq .) and piperidine ( 4 . 38 g , 44 . 3 mmoles , 1 . 2 eq . ), then stirred for 16 hours . the mixture was poured in h 2 o ( 250 ml ) and hcl 0 . 5n ( 150 ml ), and precipitation of a solid was observed . the solution was cooled to 5 °- 10 ° c . for 1 h , filtrated and dried under vacuum at 80 ° c . for 2 h . 13 g of wet solid were obtained and then crystallized from absolute ethanol , yielding 6 . 77 g of end product . 1 h - nmr , ( dmso ): 3 . 6 ppm ( s , 3h ); 3 . 8 ppm ( s ., 3h ); 6 . 5 ppm ( s ., 1h ); 6 . 9 ppm ( d ., 2h , j = 8 . 6 hz ); 10 ppm ( broad s . ); 12 . 8 ppm ( s .). the e configuration of the esocyclic double bond in position 2 was determined by means of 1d noe nmr experiments .
the use of an arylidene 2 - indolinone derivative for treating tumors involving met , pdgf - r , fgf - ri , fgf - r3 and kit tyrosine kinases , or the oncoproteins of ret family is disclosed . the invention also provides pharmaceutical compositions comprising laid arylidene 2 - indolinone derivative as well as a kit comprising , in separate containers , said derivative and an anti - cancer or anti - tumor agent .