Patent Application: US-201314430101-A

Abstract:
the present invention relates to compounds which modulate the activity of beta - hydroxylase β - hydroxylase ), including novel 2 - aryl - 5 - amino - 3 - furanone and 2 - heteroaryl - 5 - amino - 3 - furanone compounds , pharmaceutical compositions thereof , methods for their synthesis , and methods of using these compounds to modulate the activity of asph in an a cell - free sample , a cell - based assay , and in a subject . other aspects of the invention relate to use of the compounds disclosed herein to ameliorate or treat cell proliferation disorders .

Description:
asph ( a . k . a ., aah ) is a member of the α - ketoglutarate - dependent dioxygenase family enzyme . it has a predicted molecular mass of ˜ 86 kd and catalyzes the hydroxylation of specific asp ( asparate ) and asn ( asparagine ) residues in egf - like domains of certain receptor proteins such as notch . overexpression of asph has been observed in a broad range of malignant neoplasms including hepatocellular carcinoma , cholangiocellular carcinoma , pancreatic cancer , prostate cancer , breast cancer , glioblastoma , lung and colon cancer . however , asph has low or negligible expression in normal adult tissues except for proliferating trophoblastic cells of the placenta . in human hcc cell lines , asph promotes the motility and invasiveness of tumor cells through upregulation and activation of the notch signaling cascade . indeed , asph overexpression is reported to be a poor prognostic factor for patients with hcc and predicts early disease recurrence and reduced survival . especially in colon cancer , there is a significant association between poor surgical outcome and asph expression , which is considered an independent risk factor indicating poor prognosis with this disease . another tumor with high asph expression is pancreatic cancer ( pc ) which is the fourth leading cause of cancer mortality in the united states with a five - year survival rate of 5 - 6 %. pc is an extremely aggressive tumor refractory to most therapies . there is a need to define the molecular pathogenesis of pc and develop more effective treatment strategies . signaling pathways mediated by asph participate in the growth and metastasis of pc during oncogenesis . this surprising discovery on the role of asph overexpression in pc pathogenesis indicates that asph is a molecular target for therapy and that inhibition of asph in this type of cancer leads to clinical benefit . accordingly , in one aspect , the invention features an asparatyl ( asparaginyl ) beta - hydroxylase ( asph ) inhibitory compound for use in a method of reducing proliferation , migration , invasion , or metastasis of a tumor cell in the treatment of cell proliferative disorder , comprising contacting said tumor cell with the asph inhibitory compound , wherein the asph inhibitory compound is of formula ia or ib : ar 1 is substituted or unsubstituted c 6 - c 20 aryl or 5 to 20 - membered heteroaryl ; w 1 is a single bond , o , cr 50 r 51 , or nr 52 when x is co and w 1 is a single bond , cr 50 r 51 , or nr 52 when x is so 2 ; and each of r 50 , r 51 , r 52 , and r 53 independently is selected from the group consisting of hydrogen , substituted or unsubstituted c 1 - c 6 alkyl , substituted or unsubstituted c 2 - c 6 alkenyl , substituted or unsubstituted c 2 - c 6 alkynyl , substituted or unsubstituted c 6 - c 20 aryl , substituted or unsubstituted c 7 - c 26 arylalkyl , substituted or unsubstituted 5 to 20 - membered heteroaryl , and substituted or unsubstituted 6 - 26 membered heteroarylalkyl . in one embodiment , the compound for said use is of formula ia , or a salt , ester , metabolite , prodrug , or solvate thereof . the compound of formula ia may have one or more of the following features when applicable . each of ar 1 and ar 2 independently is unsubstituted c 6 - c 14 aryl , unsubstituted 5 to 14 - membered heteroaryl , or c 6 - c 14 aryl or 5 to 14 - membered heteroaryl each substituted with one or more substituents selected from the group consisting of halo , cn , no 2 , no , n 3 , or a , nr a r b , c ( o ) r a , c ( o ) or a , c ( o ) nr a r b , nr b c ( o ) r a , — s ( o ) b r a , — s ( o ) b nr a r b , or r s1 , in which r s1 is c 1 - c 6 alkyl , c 2 - c 6 alkenyl , c 2 - c 6 alkynyl , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 5 - or 6 - membered heteroaryl , or 4 to 12 - membered heterocycloalkyl , b is 0 , 1 , or 2 , each of r a and r b , independently is h or r s2 , and r s2 is c 1 - c 6 alkyl , c 2 - c 6 alkenyl , c 2 - c 6 alkynyl , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 4 to 12 - membered heterocycloalkyl , or 5 - or 6 - membered heteroaryl ; and each of r s1 and r s2 , is optionally substituted with one or more substituents selected from the group consisting of halo , oh , oxo , c ( o ) oh , c ( o ) o — c 1 - c 6 alkyl , cn , c 1 - c 6 alkyl , c 1 - c 6 alkoxyl , amino , mono - c 1 - c 6 alkylamino , di - c 1 - c 6 alkylamino , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 4 to 12 - membered heterocycloalkyl , and 5 - or 6 - membered heteroaryl . for example , r 53 is unsubstituted c 1 - c 6 alkyl or c 1 - c 6 alkyl substituted with one or more substituents selected from halo , oh , cn , and amino . for example , x is s ( o ) 2 and w 1 is cr 50 r 51 . for example , x is s ( o ) 2 and w 1 is a single bond . for example , x is c ( o ) and w 1 is o , or x is c ( s ) and w 1 is nr 52 . for example , each of r 50 , r 51 , and r 52 independently is h , unsubstituted c 1 - c 6 alkyl , or c 1 - c 6 alkyl substituted with one or more substituents selected from halo , oh , cn , and amino . for example , each of ar 1 and ar 2 independently is phenyl , naphthyl , or 5 to 10 - membered heteroaryl , each of which is optionally substituted with one or more substituents selected from the group consisting of halo , cn , no 2 , no , n 3 , or a , nr a r b , c ( o ) r a , c ( o ) or a , c ( o ) nr a r b , nr b c ( o ) r a , — s ( o ) b r a , — s ( o ) b nr a r b , or r s1 , in which r s1 is c 1 - c 6 alkyl , c 2 - c 6 alkenyl , c 2 - c 6 alkynyl , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 5 - or 6 - membered heteroaryl , or 4 to 12 - membered heterocycloalkyl , b is 0 , 1 , or 2 , each of r a and r b , independently is h or r s2 , and r s2 is c 1 - c 6 alkyl , c 2 - c 6 alkenyl , c 2 - c 6 alkynyl , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 4 to 12 - membered heterocycloalkyl , or 5 - or 6 - membered heteroaryl ; and each of r s1 and r s2 , is optionally substituted with one or more substituents selected from the group consisting of halo , oh , oxo , c ( o ) oh , c ( o ) o — c 1 - c 6 alkyl , cn , c 1 - c 6 alkyl , c 1 - c 6 alkoxyl , amino , mono - c 1 - c 6 alkylamino , di - c 1 - c 6 alkylamino , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 4 to 12 - membered heterocycloalkyl , and 5 - or 6 - membered heteroaryl . for example , each of ar 1 and ar 2 independently is phenyl , naphthyl , or 5 to 10 - membered heteroaryl , each of which is optionally substituted with one or more substituents selected from the group consisting of halo , cn , no 2 , no , n 3 , or a , nr a r b , c ( o ) r a , c ( o ) or a , or r s1 , in which r s1 is c 1 - c 6 alkyl , each of r a and r b , independently is h or r s2 , and r s2 is c 1 - c 6 alkyl ; and each of r s1 and r s2 , is optionally substituted with one or more substituents selected from the group consisting of halo , oh , c 1 - c 6 alkoxyl , amino , mono - c 1 - c 6 alkylamino , and di - c 1 - c 6 alkylamino . for example , each of ar 1 and ar 2 independently is selected from phenyl , 1 - naphthyl , 2 - naphthyl , 2 - furanyl , 2 - thiazolyl , 2 - pyridyl , 3 - pyridyl , 4 - pyridyl , 2 - quinolinyl , 3 - quinolinyl , 4 - quinolinyl , 2 - chlorophenyl , 3 - chlorophenyl , 4 - chlorophenyl , 2 - fluorophenyl , 3 - fluorophenyl , 4 - fluorophenyl , 2 - trifluoromethylphenyl , 3 - trifluoromethylphenyl , 4 - trifluoromethylphenyl , 2 - cyanophenyl , 3 - cyanophenyl , 4 - cyanophenyl , 3 - carboxymethylphenyl , 2 - methoxyphenyl , 3 - methoxyphenyl , 4 - methoxyphenyl , 2 , 3 - dichlorophenyl , 2 , 4 - dichlorophenyl , 2 , 5 - dichlorophenyl , 3 , 4 - dichlorophenyl , 3 , 5 - dichlorophenyl , 2 , 3 - difluorophenyl , 2 , 4 - difluorophenyl , 2 , 5 - difluorophenyl , 3 , 4 - difluorophenyl , 3 , 5 - difluorophenyl , 2 , 3 - dimethoxyphenyl , 2 , 4 - dimethoxyphenyl , 2 , 5 - dimethoxyphenyl , 3 , 4 - dimethoxyphenyl , 3 , 5 - dimethoxyphenyl , 2 - chloro - 6 - fluorophenyl , 3 - chloro - 4 - fluorophenyl , 2 - chloro - 4 - fluorophenyl , 4 - chloro - 3 - fluorophenyl , 3 - chloro - 2 - fluorophenyl , 2 - chloro - 5 - fluorophenyl , 4 - chloro - 2 - fluorophenyl , and 5 - chloro - 2 - fluorophenyl . in another embodiment , the compound for said use is of formula ib , or a salt , ester , metabolite , prodrug , or solvate thereof . the compound of formula ib may have one or more of the following features when applicable . for example , r 53 is unsubstituted c 1 - c 6 alkyl or c 1 - c 6 alkyl substituted with one or more substituents selected from halo , oh , cn , and amino . for example , x is s ( o ) 2 and w 1 is cr 50 r 51 . for example , x is s ( o ) 2 and w 1 is a single bond . for example , x is c ( o ) and w 1 is o , or x is c ( s ) and w 1 is nr 52 . for example , each of r 50 , r 51 , and r 52 independently is h , unsubstituted c 1 - c 6 alkyl , or c 1 - c 6 alkyl substituted with one or more substituents selected from halo , oh , cn , and amino . for example , ar 1 is phenyl , naphthyl , or 5 to 10 - membered heteroaryl , each of which is optionally substituted with one or more substituents selected from the group consisting of halo , cn , no 2 , no , n 3 , or a , nr a r b , c ( o ) r a , c ( o ) or a , c ( o ) nr a r b , nr b c ( o ) r a , — s ( o ) b r a , — s ( o ) b nr a r b , or r s1 , in which r s1 is c 1 - c 6 alkyl , c 2 - c 6 alkenyl , c 2 - c 6 alkynyl , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 5 - or 6 - membered heteroaryl , or 4 to 12 - membered heterocycloalkyl , b is 0 , 1 , or 2 , each of r a and r b , independently is h or r s2 , and r s2 is c 1 - c 6 alkyl , c 2 - c 6 alkenyl , c 2 - c 6 alkynyl , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 4 to 12 - membered heterocycloalkyl , or 5 - or 6 - membered heteroaryl ; and each of r s1 and r s2 , is optionally substituted with one or more substituents selected from the group consisting of halo , oh , oxo , c ( o ) oh , c ( o ) o — c 1 - c 6 alkyl , cn , c 1 - c 6 alkyl , c 1 - c 6 alkoxyl , amino , mono - c 1 - c 6 alkylamino , di - c 1 - c 6 alkylamino , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 4 to 12 - membered heterocycloalkyl , and 5 - or 6 - membered heteroaryl . for example , ar 1 is phenyl , naphthyl , or 5 to 10 - membered heteroaryl , each of which is optionally substituted with one or more substituents selected from the group consisting of halo , cn , no 2 , no , n 3 , or a , nr a r b , c ( o ) r a , c ( o ) or a , or r s1 , in which r s1 is c 1 - c 6 alkyl , each of r a and r b , independently is h or r s2 , and r s2 is c 1 - c 6 alkyl ; and each of r s1 and r s2 , is optionally substituted with one or more substituents selected from the group consisting of halo , oh , c 1 - c 6 alkoxyl , amino , mono - c 1 - c 6 alkylamino , and di - c 1 - c 6 alkylamino . for example , ar 1 is selected from phenyl , 1 - naphthyl , 2 - naphthyl , 2 - furanyl , 2 - thiazolyl , 2 - pyridyl , 3 - pyridyl , 4 - pyridyl , 2 - quinolinyl , 3 - quinolinyl , 4 - quinolinyl , 2 - chlorophenyl , 3 - chlorophenyl , 4 - chlorophenyl , 2 - fluorophenyl , 3 - fluorophenyl , 4 - fluorophenyl , 2 - trifluoromethylphenyl , 3 - trifluoromethylphenyl , 4 - trifluoromethylphenyl , 2 - cyanophenyl , 3 - cyanophenyl , 4 - cyanophenyl , 3 - carboxymethylphenyl , 2 - methoxyphenyl , 3 - methoxyphenyl , 4 - methoxyphenyl , 2 , 3 - dichlorophenyl , 2 , 4 - dichlorophenyl , 2 , 5 - dichlorophenyl , 3 , 4 - dichlorophenyl , 3 , 5 - dichlorophenyl , 2 , 3 - difluorophenyl , 2 , 4 - difluorophenyl , 2 , 5 - difluorophenyl , 3 , 4 - difluorophenyl , 3 , 5 - difluorophenyl , 2 , 3 - dimethoxyphenyl , 2 , 4 - dimethoxyphenyl , 2 , 5 - dimethoxyphenyl , 3 , 4 - dimethoxyphenyl , 3 , 5 - dimethoxyphenyl , 2 - chloro - 6 - fluorophenyl , 3 - chloro - 4 - fluorophenyl , 2 - chloro - 4 - fluorophenyl , 4 - chloro - 3 - fluorophenyl , 3 - chloro - 2 - fluorophenyl , 2 - chloro - 5 - fluorophenyl , 4 - chloro - 2 - fluorophenyl , and 5 - chloro - 2 - fluorophenyl . in certain embodiments , said cell proliferative disorder comprises pancreatic cancer , hepatocellular cancer , cholangiocarcinoma , lung cancer , colon cancer , breast cancer , prostatic cancer , and glioblastoma . in one embodiment , said compound is administered intravenously , orally , or subcutaneously . in one embodiment , said compound is administered at a dose of 0 . 01 to 50 milligrams / kilogram of body weight . asph is a highly conserved cell - surface protein in hepatocellular carcinoma ( hcc ). both the liver and the pancreas are derived from an early progenitor cell type and asph is expressed in embryo but not in adult tissues . asph re - expression was observed in human pc tissue microarrays by immunohistochemical staining ( ihs ) as shown in fig1 a - e . high level cell surface localization of asph was present in 101 of 104 ( 97 %) pancreatic ductal adenocarcinoma with negligible expression in normal pancreas , and other adult human tissues . asph enhances cell migration , invasion , and metastasis in hcc and also pc . activation of notch signaling by asph is a final effector mechanism responsible for generation of this highly aggressive and malignant phenotype . the regulation , expression , and function of asph has been observed in many tumors ( u . s . pat . nos . 6 , 835 , 370 ; 7 , 094 , 556 ; 6 , 812 , 206 ; 6 , 815 , 415 ; 6 , 797 , 696 ; 6 , 783 , 758 ; and u . s . published patent application no . 2005 - 0123545 ; hereby incorporated by reference ) and asph has been found to be overexpressed in pancreatic ductal adenocarcinoma ( pc ) indicating that it is a therapeutic target for treatment of pc . asph catalyzes post - translational hydroxylation of β - carbons of specific aspartate and asparaginyl residues in epidermal growth factor ( egf )- like domains residing in proteins such as notch and jagged ( jag ) which are involved in cell growth , differentiation , cellular migration , adhesion , and motility . the catalytic activity resides in the c - terminus and is conferred by the 675 his residue ; mutation to an alanine abolishes asph enzymatic and transforming activity . asph is overexpressed in tumors derived from the endoderm such as liver , pancreas , colon and lung , and translocates from the endoplasmic reticulum ( er ) to the plasma membrane where it becomes accessible to the extracellular environment . it has negligible to very low expression in normal human tissue with the notable exception of the placenta which is an invasive tissue , and its expression there is robust . compounds are administered directly into a tumor site or systemically to inhibit asph hydroxylase activity . methods of inhibiting tumor growth also include administering a compound which inhibits haah hydroxylation of a notch polypeptide . for example , the compound inhibits hydroxylation of an egf - like cysteine - rich repeat sequence in a notch polypeptide , e . g ., one containing the consensus sequence polypeptides containing an egf - like cysteine - rich repeat sequence are administered to block hydroxylation of endogenous notch . asph is expressed in many organs during embryogenesis presumably to promote cell motility and migration for cell patterning and organ development ; its expression is “ shut off ” in the adult only to re - emerge during oncogenesis where its function may be required for generation of malignant phenotypes . it appears not to be overexpressed during cell proliferation ; however , there is low - level expression in dysplastic ductal cells of pancreatic intraepithelial lesions ( panins ) as well as dysplastic hepatocytes in hepatitis b ( hbv ) and c ( hcv ) infected liver . transcriptional regulation of asph is provided by tripartite signaling pathways in / igf1 / irs1 / mapk / erk , in / igf1 / irs1 / pi3k / akt , and wnt / β - catenin . post - transcriptional regulation of asph is mediated by phosphorylation of gsk - 3β - related motifs located in the n - terminal region of the molecule . one mechanism by which asph exerts its effector function is by activating downstream notch signaling to promote cell migration and invasion . table 1 details the overexpression of asph at the protein and rna level as determined by ihs and qrt - pcr respectively in various human tumors indicating that it is a therapeutic target for a variety of human solid malignancies with a poor prognosis . fig1 and 2 show examples of asph protein expression by ihs . the c - terminus of asph contains amino acid ( aa ) sequence of the catalytic site ( m 670 hpgfh 675 ) and its sequence is identical in human , rat , mouse , and cattle . the h 675 aa is specifically involved in fe 2 + coordination and critical for its enzymatic activity , also highly conserved in the chicken and fly . a h 675 r mutation reduces β - hydroxylase activity to & lt ; 1 % of wild - type protein while h 675 d reduces it to 20 %. in this context the h 675 r mutant protein loses the ability to promote cell proliferation , motility , migration , invasion , colony formation in soft agar , as well as metastasis and tumor formation in nude mice compared to the “ wild - type ” sequence and it also can function as a dominant negative mutant to inhibit the function of the endogenous “ wild - type ” protein . these findings indicate that inhibition of β - hydroxylase activity promotes anti - tumor effects . the crystal structure of the catalytic site region has been elucidated and is available in the public database ( rcsb protein database ; code 3rcq ). small molecule inhibitors for use as anti - tumor agents were identified by their ability to fit into the catalytic site region of asph and inhibit asph enzymatic activity . compounds of the present invention can be prepared in a variety of ways using commercially available starting materials , compounds known in the literature , or from readily prepared intermediates , by employing standard synthetic methods and procedures either known to those skilled in the art , or which will be apparent to the skilled artisan in light of the teachings herein . standard synthetic methods and procedures for the preparation of organic molecules and functional group transformations and manipulations can be obtained from the relevant scientific literature or from standard textbooks in the field . although not limited to any one or several sources , classic texts such as smith , m . b ., march , j ., march &# 39 ; s advanced organic chemistry : reactions , mechanisms , and structure , 5 th edition , john wiley & amp ; sons : new york , 2001 ; greene , t . w ., wuts , p . g . m ., protective groups in organic synthesis , 3 rd edition , john wiley & amp ; sons : new york , 1999 ; r . larock , comprehensive organic transformations , vch publishers ( 1989 ); l . fieser and m . fieser , fieser and fieser &# 39 ; s reagents for organic synthesis , john wiley and sons ( 1994 ); and l . paquette , ed ., encyclopedia of reagents for organic synthesis , john wiley and sons ( 1995 ), incorporated by reference herein , are useful and recognized reference textbooks of organic synthesis known to those in the art . the following descriptions of synthetic methods are designed to illustrate , but not to limit , general procedures for the preparation of compounds of the present invention . in the reaction schemes described herein , multiple stereoisomers may be produced . when no particular stereoisomer is indicated , it is understood to mean all possible stereoisomers that could be produced from the reaction . a person of ordinary skill in the art will recognize that the reactions can be optimized to give one isomer preferentially , or new schemes may be devised to produce a single isomer . if mixtures are produced , techniques such as preparative thin layer chromatography , preparative hplc , preparative chiral hplc , or preparative sfc may be used to separate the isomers . scheme 1 above shows the synthetic strategies for compounds of formula ia . reactions a - f are as follows : ( a ) kcn , glyoxal , na 2 co 3 , h 2 o ; ( b ) clso 2 r , tea , thf ; ( c ) clso 2 ch 2 ph , tea , thf ; ( d ) clso 2 ch 2 ar 2 , tea , thf ; ( e ) clco 2 ar 2 , tea , thf ; ( f ) ar 2 ncs , na 2 co 3 , h 2 o . based on the crystal structure of the asph catalytic site , computer generated drug design was performed that has led to the synthesis of a series of parent compounds and derivatives to fit into the pocket of the catalytic site and inhibit the β - hydroxylase activity . parent compounds ( compound 307 { 7c or mo - i - 1000 }, and compound 310 { 8c or mo - i - 1100 }) were synthesized and examined for inhibition of β - hydroxylase activity using a high throughput screening assay . synthesis of asph inhibitors was accomplished in two steps . the first step was a three component reaction including an aromatic aldehyde , glyoxal bisulfate addition product , and potassium cyanide to yield an arylhydroxytetronimide . in the second step the arylhydroxytetronimide was sulfonylated with phenylmethanesulfonyl chloride in dry tetrahydrofuran to yield compounds of formula ia ( scheme 1 ). compounds were characterized by 1h and 13c nuclear magnetic resonance , high resolution mass spectroscopy , high performance liquid chromatography , infra - red spectroscopy , melting point , elemental analysis and binding to asph by isothermal titration calorimetry . compound 403 { 3c or mo - i - 1000 }, compound 310 { 8c or mo - i - 1100 }, compound 404 { 4c or mo - i - 400 } and compound 405 { 5c or mo - i - 500 } were identified as hits . this led to identification of a mixture of enantiomers as the lead compounds 2 and 3 and the additional recognition that the compound 405 { 5c or mo - i - 500 } compound demonstrated biologic activity through its action on asph enzymatic activity . egf and egf - like domains are well known in the art and generally include six cysteine residues which have been shown ( in egf ) to be involved in disulfide bonds . the main structure is a two - stranded beta - sheet followed by a loop to a c - terminal short two - stranded sheet . subdomains between the conserved cysteines vary in length . examples include those described in davis , c g , 1990 , new biol . 2 ( 5 ): 410 - 9 ; blomquist et al ., 1984 , proc natl acad sci usa . 81 ( 23 ): 7363 - 7 ; hommel et al ., 1002 , j mol biol . 227 ( 1 ): 271 - 82 ; doolittle et al ., 1984 , nature . 307 ( 5951 ): 558 - 60 ; appella et al ., 1988 , febs lett . 231 ( 1 ): 1 - 4 ; sorkin a ., 2001 , biochem soc trans . aug ; 29 ( pt 4 ): 480 - 4 ; each of which is hereby incorporated by reference ). fig3 a - c represents the strategy for development and characterization of the performance of an assay to measure asph enzymatic activity . fig3 a describes the biochemical reaction catalyzed by asph . fig3 b depicts the read - out of the assay . protein lysates were extracted from focus cells ( high level of asph cell surface expression ) treated with 1 - 10 μm of each parent compound for 24 hours . the lysates were added into 96 well - plates coated with asph monoclonal antibodies ( mabs ) to add an antigen specific ( asph ) capture step to the assay design . after incubation and washing , only asph is captured on each well . the reaction is carried out with 60 μm of an egf domain containing 39aa peptide , 100 μm fecl 2 and 40 μm 14 c labeled α - ketoglutarate were added into each well . the 14 co 2 was captured on a glass fiber membrane soaked in ca ( oh ) 2 . radioactivity captured was quantified by a phosphor - imager as shown in fig3 c . fig3 d shows that compound 310 { 8c or mo - i - 1100 } at a concentration of 1 μm substantially inhibits β - hydroxylase activity . the compound was further characterized for clinical use as an anti - tumor agent for pc and hcc and other asph - expressing tumors . after demonstrating that compound 310 { 8c or mo - i - 1100 } inhibited β - hydroxylase activity using the asph specific mab capture enzymatic assay , its activity as an anti - tumor agent in a nude mouse model of subcutaneous ( s . c .) tumor growth was evaluated . the hpafii aspc - 1 human pc cell line which expresses a high level of asph was implanted ( 5 × 10 6 cells s . c .) into the back of nude mice . tumors were allowed to grow to approximately 100 mm 3 after one week , and compound 310 { 8c or mo - i - 1100 } was then administered intraperitoneal ( i . p .) at a concentration of 50 mg / kg . the treatment regimen as shown in fig4 a - b included i . p . injections on a daily basis for five days followed by every other day until the experiment was terminated due to the large size of tumors observed in the control group that received a dmso vehicle injection . fig4 a - b demonstrates the growth rate and substantial inhibition of hpafii tumor formation over the course of the study . there were 15 nude mice in each group ( control vs . treatment ). therefore , this study demonstrates that the compound ( compound 310 { 8c or mo - i - 1100 }) was active in vitro inhibiting asph β - hydroxylase activity , and performed well in vivo as an anti - tumor agent . in vitro effects of asph inhibitors for cell proliferation and metabolism : a mtt assay was performed to evaluate the effect of compound 310 { 8c or mo - i - 1100 } on cell proliferation and viability . mtt is reduced to purple formazan in living cells . focus cells were used as a high asph expressing human hcc cell line . the results shows that treatments with compound 310 { 8c or mo - i - 1100 } for 24 hours dose - dependently decreased the od ( optical density ) value in focus cells ( fig5 a - b ). however , in nih - 3t3 cells , which is a mouse embryo fibroblast cell line not expressing asph , compound 310 { 8c or mo - i - 1100 } had no effect on the od value of mtt indicating that compound 310 { 8c or mo - i - 1100 } is highly specific for the β - hydroxylase activity of asph and did not affect cells that lacked asph expression . the mtt assay measures cellular metabolic activity via nad ( p ) h - dependent cellular oxidoreductase enzymes . however , as shown in fig5 c , compound 310 { 8c or mo - i - 1100 } also decreased cell viability in human hcc cell lines at 5 μm ( focus , hep3b , hepg2 and huh7 ) which express asph ( inhibition rate 37 . 9 %, 60 %, 59 % and 50 %, respectively ) but not nih 3t3 cells with no expression of asph . in order to evaluate the effect of long - term exposure of compound 310 { 8c or mo - i - 1100 }, the colony formation assay ( an assay of malignant potential and phenotype ) in which cells were treated with inhibitor for 3 weeks was performed . treatment with compound 310 { 8c or mo - i - 1100 } resulted in reduced colony formation at 5 tm concentration ( inhibition rate 36 . 8 %) ( fig6 a - b ). we performed similar studies using the compound 405 { 5c or mo - i - 500 } compound . similar to the findings in compound 310 { 8c or mo - i - 1100 }, we observed that compound 405 { 5c or mo - i - 500 } at 5 μm also inhibited asph enzymatic activity as measured by the 14 co 2 - ketoglutarate - dependent capture assay . the structure of compound 405 { 5c or mo - i - 500 } is quite different than compound 310 { 8c or mo - i - 1100 } and the results of the biological activity of compound 405 { 5c or mo - i - 500 } is shown in fig7 a - b . additional experiments measured the in vitro effects of compound 405 { 5c or mo - i - 500 } on cell proliferation and metabolism as shown in fig8 a - d . in contrast to compound 310 { 8c or mo - i - 1100 } effects , compound 405 { 5c or mo - i - 500 } has striking effects on cell proliferation and metabolic activity at micromolar concentrations in both focus cells ( which contain high levels of asph cell surface expression ) and nih - 3t3 cells which do not . these findings indicate that compound 405 { 5c or mo - i - 500 } has hydroxylase inhibitory activity but also probably inhibits other hydroxylases or proteins that may be important for cell viability and growth as well since nih 3t3 cells lacking asph were susceptible to its inhibitory effects . more striking was the colony formation assay that showed that 1 . 25 μm concentrations of compound 405 { 5c or mo - i - 500 } had substantial inhibitory effects on colony formation of focus hcc cells indicating high potency in this assay of cellular transformation . in summary both compound 310 { 8c or mo - i - 1100 } and compound 405 { 5c or mo - i - 500 } inhibit asph β - hydroxylase activity , cell viability and metabolism , as well as colony formation in soft agar but compound 310 { 8c or mo - i - 1100 } is more highly specific for the β - hydroxylase of asph as shown in fig5 a - c . the effect of compound 310 { 8c or mo - i - 1100 } on anchorage independent growth in soft agar was evaluated . the ability of forming colonosphere in soft agar is considered to be a rigid test for tumorigenic potential . asph expression results in cells acquiring the ability to form colonospheres in soft agar . these results indicated that asph plays a key role in establishing tumor , growth , invasion and metastasis in vivo . in order to evaluate the effect of asph inhibitor for anchorage independent colony formation , focus cells were incubated in soft agar with or without this asph enzymatic inhibitor . after 3 weeks incubation , compound 310 { 8c or mo - i - 1100 } reduced colonosphere formation of focus cells ( fig9 a ). as shown in fig9 b , c treatment with compound 310 { 8c or mo - i - 1100 } produced a dose - dependent and highly significant reduction both in number and size of the colonies after 3 weeks of culture . this assay is a standard method to monitor malignant growth that reflects the ability to form tumors that grow aggressively in vivo . these results confirm that compound 310 { 8c or mo - i - 1100 } impairs the generation of a malignant phenotype in vitro . the effect of compound 405 { 5c or mo - i - 500 } on anchorage independent cell growth was evaluated as shown in fig1 a - b . it was striking that 1 μm exposure of hcc cells grown in soft agar dramatically reduced the number of colonies formed , and the colony size in a dose - dependent manner . thus , compound 405 { 5c or mo - i - 500 } had very similar but more potent effects on anchorage - independent cell growth as did compound 310 { 8c or mo - i - 1100 } for this assay of cellular transformation that correlates well with tumor formation in vivo . in vitro effects of asph inhibitors for cell motility and invasiveness in human hcc cell : the β - hydroxylase activity is required for asph to mediate its effects on cell motility . directional motility was measured using boyden chamber - type culture inserts equipped with porous membranes . focus cells were pretreated with 5 μm of compound 310 { 8c or mo - i - 1100 } and compound 405 { sc or mo - i - 500 } β - hydroxylase inhibitor for 24 hours , and then placed into the upper chamber . migration was allowed to proceed for 30 min . atplite was used to quantify viable cell density . the cells in the upper well and upper surface of membrane reflects the number of non - migrating cells , luminescence measured at the bottom surface of the membrane reflects the number of migrating and non - adherent cells and luminescence measured in the bottom well was reflect migrating and non - adherent cells . as shown in fig1 a - c total migrated cells were reduced following compound 310 { 8c or mo - i - 1100 } treatment but the population of non - adherent cells was unchanged ; the migrating and adherent cells were significantly reduced . cell invasiveness was assessed by invasion assay using matrigel coated membrane , in which cells were allowed to proceed for 6 hours ; those found adhered on the bottom surface of membrane were regarded as invading cells . compound 310 { 8c or mo - i - 1100 } treatment focus cells significantly reduced invasiveness compared to cells incubated with dmso as a control ( fig1 d ). results demonstrated that compound 310 { 8c or mo - i - 1100 } inhibited cell motility , migration and invasiveness of human hcc cells . in vivo effects of asph inhibitors on cell motility and invasiveness were evaluated using the human hcc focus cell line . as shown in fig1 a - b , compound 405 { 5c or mo - i - 500 } had a pronounced effect on cell motility and invasion as well . note that compound 405 { 5c or mo - i - 500 } was slightly more potent inhibiting cell motility and invasion than compound 310 { 8c or mo - i - 1100 }, but both were highly active in these two assays that characterize the malignant phenotype . thus , these small molecule inhibitors of asph - β - hydroxylase have a profound effect on the function of the metastatic phenotype by reducing the ability of tumor cells to migrate and invade , and thus substantially alter their biologic function and metastatic potential . in vivo effects of an asph inhibitor , compound 310 { 8c or mo - i - 1100 }, on subcutaneous xenograft development and growth of human hepatocellular carcinoma . the human hcc cell line focus is known to be a highly aggressive tumor forming cell line in vivo . to investigate in vivo anti - tumor efficacy of the asph inhibitor , compound 310 { 8c or mo - i - 1100 } at 20 mg / kg per day was administered on 5 consecutive days for 2 weeks and every other day thereafter . as shown in fig1 a - b , the administration of compound 310 { 8c or mo - i - 1100 } significantly reduced hcc subcutaneous xenograft growth . the mean tumor volumes were substantially decreased by treatment with compound 310 { 8c or mo - i - 11001 } on day 12 following treatment , and tumor volumes in treated mice were reduced an average of 31 . 7 % compared to those observed in control mice ( fig1 b ). none of the compound 310 { 8c or mo - i - 1100 } treated mice showed signs of wasting or other adverse effects relative to control mice . thus , compound 310 { 8c or mo - i - 1100 } was tolerated well at this dose level where striking antitumor efficacy was observed . thus , a specific β - hydroxylase inhibitor such as compound 310 { 8c or mo - i - 1100 } substantially reduced tumor growth of hcc as shown in fig1 a - b but also inhibited the development and growth of pancreatic cancer as well ( fig6 a - b ) since the tumor also has high level expression of asph . such findings indicate that any asph expressing human tumor are responsive to these specific β - hydroxylase inhibitors . these asph inhibitor compounds represent a class of anti - tumor agents that has substantial anti - tumor activity against a large number of solid human tumors known to have a dismal prognosis . asph is overexpressed on the cell surface of human tumor cells within solid tumors and has low or negligible expression in normal human tissues . asph expression is present in most , if not all , tumor cells . asph is also exposed to the extracellular environment which makes it an excellent therapeutic target since it has easy access to small molecule inhibitors of the catalytic activity through the blood . the data described herein support the following conclusions : asph overexpression causes increased motility , migration , invasion and metastasis of hcc cells as well as other human tumor cell lines ; many human solid tumors with a dismal prognosis overexpress asph on the cell surface including but not limited to pancreatic , hepatocellular , cholangio -, colon , breast , prostate , lung , and glioblastoma cancers ; the catalytic site and enzymatic activity are critical for asph mediated malignant transformation , and the subsequent generation of an invasive and metastatic tumor phenotype ; asph exerts its biologic effects on increased migration , invasion , and metastasis of tumor cells by activation of notch signaling cascade ; small molecule inhibitors of the β - hydroxylase activity have been discovered based on the crystal structure of the c - terminal catalytic site of asph ; tumor cells exposed to these inhibitors such as compound 310 { 8c or mo - i - 1100 } and compound 405 { 5c or mo - i - 500 } have reduced proliferation , migration , invasion , and colony formation in soft agar which impairs the ability of these cells to grow and metastasize ; these small molecule inhibitors of asph enzymatic activity have striking and unexpected anti - tumor effects in vivo in animal models of human pancreatic and liver cancer growth and development . thus , these studies demonstrate that compounds which specifically inhibit the β - hydroxylase activity are useful to reduce the growth and / or inhibit metastases of asph expressing human solid tumors , in particular those known to have a dismal clinical prognosis , e . g ., pancreatic cancer , hepatocellular cancer , cholangiocarcinoma , lung , colon cancer , breast cancer , prostatic cancer , and glioblastoma . in another aspect , this invention features a compound of formula ia : ar 1 is substituted or unsubstituted c 6 - c 20 aryl or 5 to 20 - membered heteroaryl ; w 1 is a single bond , o , cr 50 r 51 , or nr 52 when x is co and w 1 is a single bond , cr 50 r 51 , or nr 52 when x is so 2 ; and each of r 50 , r 51 , r 52 , and r 53 independently is selected from the group consisting of hydrogen , substituted or unsubstituted c 1 - c 6 alkyl , substituted or unsubstituted c 2 - c 6 alkenyl , substituted or unsubstituted c 2 - c 6 alkynyl , substituted or unsubstituted c 6 - c 20 aryl , substituted or unsubstituted c 7 - c 26 arylalkyl , substituted or unsubstituted 5 to 20 - membered heteroaryl , and substituted or unsubstituted 6 - 26 membered heteroarylalkyl , provided that when ar 1 is 4 - chlorophenyl , then r 53 is not methyl or unsubstituted phenyl . the compound of formula ia may have one or more of the following features when applicable . each of ar 1 and ar 2 independently is unsubstituted c 6 - c 14 aryl , unsubstituted 5 to 14 - membered heteroaryl , or c 6 - c 14 aryl or 5 to 14 - membered heteroaryl each substituted with one or more substituents selected from the group consisting of halo , cn , no 2 , no , n 3 , or a , nr a r b , c ( o ) r a , c ( o ) or a , c ( o ) nr a r b , nr b c ( o ) r a , — s ( o ) b r a , — s ( o ) b nr a r b , or r s1 , in which r s1 is c 1 - c 6 alkyl , c 2 - c 6 alkenyl , c 2 - c 6 alkynyl , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 5 - or 6 - membered heteroaryl , or 4 to 12 - membered heterocycloalkyl , b is 0 , 1 , or 2 , each of r a and r b , independently is h or r s2 , and r s2 is c 1 - c 6 alkyl , c 2 - c 6 alkenyl , c 2 - c 6 alkynyl , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 4 to 12 - membered heterocycloalkyl , or 5 - or 6 - membered heteroaryl ; and each of r s1 and r s2 , is optionally substituted with one or more substituents selected from the group consisting of halo , oh , oxo , c ( o ) oh , c ( o ) o — c 1 - c 6 alkyl , cn , c 1 - c 6 alkyl , c 1 - c 6 alkoxyl , amino , mono - c 1 - c 6 alkylamino , di - c 1 - c 6 alkylamino , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 4 to 12 - membered heterocycloalkyl , and 5 - or 6 - membered heteroaryl . for example , r 53 is unsubstituted c 1 - c 6 alkyl or c 1 - c 6 alkyl substituted with one or more substituents selected from halo , oh , cn , and amino . for example , x is s ( o ) 2 and w 1 is cr 50 r 51 . for example , x is s ( o ) 2 and w 1 is a single bond . for example , x is c ( o ) and w 1 is o , or x is c ( s ) and w 1 is nr 52 . for example , each of r 50 , r 51 , and r 52 independently is h , unsubstituted c 1 - c 6 alkyl , or c 1 - c 6 alkyl substituted with one or more substituents selected from halo , oh , cn , and amino . for example , each of ar 1 and ar 2 independently is phenyl , naphthyl , or 5 to 10 - membered heteroaryl , each of which is optionally substituted with one or more substituents selected from the group consisting of halo , cn , no 2 , no , n 3 , or a , nr a r b , c ( o ) r a , c ( o ) or a , c ( o ) nr a r b , nr b c ( o ) r a , — s ( o ) b r a , — s ( o ) b nr a r b , or r s1 , in which r s1 is c 1 - c 6 alkyl , c 2 - c 6 alkenyl , c 2 - c 6 alkynyl , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 5 - or 6 - membered heteroaryl , or 4 to 12 - membered heterocycloalkyl , b is 0 , 1 , or 2 , each of r a and r b , independently is h or r s2 , and r s2 is c 1 - c 6 alkyl , c 2 - c 6 alkenyl , c 2 - c 6 alkynyl , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 4 to 12 - membered heterocycloalkyl , or 5 - or 6 - membered heteroaryl ; and each of r s1 and r s2 , is optionally substituted with one or more substituents selected from the group consisting of halo , oh , oxo , c ( o ) oh , c ( o ) o — c 1 - c 6 alkyl , cn , c 1 - c 6 alkyl , c 1 - c 6 alkoxyl , amino , mono - c 1 - c 6 alkylamino , di - c 1 - c 6 alkylamino , c 3 - c 8 cycloalkyl , c 6 - c 10 aryl , 4 to 12 - membered heterocycloalkyl , and 5 - or 6 - membered heteroaryl . for example , each of ar 1 and ar 2 independently is phenyl , naphthyl , or 5 to 10 - membered heteroaryl , each of which is optionally substituted with one or more substituents selected from the group consisting of halo , cn , no 2 , no , n 3 , or a , nr a r b , c ( o ) r a , c ( o ) or a , or r s1 , in which r s1 is c 1 - c 6 alkyl , each of r a and r b , independently is h or r s2 , and r s2 is c 1 - c 6 alkyl ; and each of r s1 and r s2 , is optionally substituted with one or more substituents selected from the group consisting of halo , oh , c 1 - c 6 alkoxyl , amino , mono - c 1 - c 6 alkylamino , and di - c 1 - c 6 alkylamino . for example , each of ar 1 and ar 2 independently is selected from phenyl , 1 - naphthyl , 2 - naphthyl , 2 - furanyl , 2 - thiazolyl , 2 - pyridyl , 3 - pyridyl , 4 - pyridyl , 2 - quinolinyl , 3 - quinolinyl , 4 - quinolinyl , 2 - chlorophenyl , 3 - chlorophenyl , 4 - chlorophenyl , 2 - fluorophenyl , 3 - fluorophenyl , 4 - fluorophenyl , 2 - trifluoromethylphenyl , 3 - trifluoromethylphenyl , 4 - trifluoromethylphenyl , 2 - cyanophenyl , 3 - cyanophenyl , 4 - cyanophenyl , 3 - carboxymethylphenyl , 2 - methoxyphenyl , 3 - methoxyphenyl , 4 - methoxyphenyl , 2 , 3 - dichlorophenyl , 2 , 4 - dichlorophenyl , 2 , 5 - dichlorophenyl , 3 , 4 - dichlorophenyl , 3 , 5 - dichlorophenyl , 2 , 3 - difluorophenyl , 2 , 4 - difluorophenyl , 2 , 5 - difluorophenyl , 3 , 4 - difluorophenyl , 3 , 5 - difluorophenyl , 2 , 3 - dimethoxyphenyl , 2 , 4 - dimethoxyphenyl , 2 , 5 - dimethoxyphenyl , 3 , 4 - dimethoxyphenyl , 3 , 5 - dimethoxyphenyl , 2 - chloro - 6 - fluorophenyl , 3 - chloro - 4 - fluorophenyl , 2 - chloro - 4 - fluorophenyl , 4 - chloro - 3 - fluorophenyl , 3 - chloro - 2 - fluorophenyl , 2 - chloro - 5 - fluorophenyl , 4 - chloro - 2 - fluorophenyl , and 5 - chloro - 2 - fluorophenyl . for example , the compound of formula ia or iia is an asph inhibitory compound . in some aspects , this invention provides for the use of a compound as herein described , or its isomer , metabolite , tautomer , pharmaceutically acceptable salt , pharmaceutical product , polymorph , crystal , n - oxide , hydrate , or any combination thereof , for treating , suppressing , preventing , reducing the severity , reducing the risk , or inhibiting a cell proliferation disorder in a subject . related aspects of the invention are directed to compositions , including pharmaceutical compositions , comprising the compounds of the invention , noted above . one aspect of the invention is directed to a pharmaceutical composition comprising at least one pharmaceutically acceptable excipient and a therapeutically effective amount of the compound or salt disclosed above . still another aspect of the invention relates to a method for pharmaceutical formulation of previously described compounds for use in oral and intravenous applications , and in implantable materials . another aspect of the present invention relates to a pharmaceutical composition including a pharmaceutical composition can contain one or more of the above - identified compounds of the present invention . typically , the pharmaceutical composition of the present invention will include a compound of the present invention or its pharmaceutically acceptable salt , as well as a pharmaceutically acceptable carrier . the term “ pharmaceutically acceptable carrier ” refers to any suitable adjuvants , carriers , excipients , or stabilizers , and can be in solid or liquid form such as , tablets , capsules , powders , solutions , suspensions , emulsions , or implantable disc . typically , the composition will contain from about 0 . 01 to 99 percent , preferably from about 20 to 75 percent of active compound ( s ), together with the adjuvants , carriers and / or excipients . while individual needs may vary , determination of optimal ranges of effective amounts of each component is within the skill of the art . typical dosages comprise about 0 . 01 to about 100 mg / kg body wt . the preferred dosages comprise about 0 . 1 to about 100 mg / kg body wt . the most preferred dosages comprise about 1 to about 100 mg / kg body wt . treatment regimen for the administration of the compounds of the present invention can also be determined readily by those with ordinary skill in art . that is , the frequency of administration and size of the dose can be established by routine optimization , preferably while minimizing any side effects . the solid unit dosage forms can be of the conventional type . the solid form can be a capsule and the like , such as an ordinary gelatin type containing the compounds of the present invention and a carrier , for example , lubricants and inert fillers such as , lactose , sucrose , or cornstarch . in another embodiment , these compounds are tabulated with conventional tablet bases such as lactose , sucrose , or cornstarch in combination with binders like acacia , cornstarch , or gelatin , disintegrating agents , such as cornstarch , potato starch , or alginic acid , and a lubricant , like stearic acid or magnesium stearate . the tablets , capsules , and the like can also contain a binder such as gum tragacanth , acacia , com starch , or gelatin ; excipients such as dicalcium phosphate ; a disintegrating agent such as com starch , potato starch , alginic acid ; a lubricant such as magnesium stearate ; and a sweetening agent such as sucrose , lactose , or saccharin . when the dosage unit form is a capsule , it can contain , in addition to materials of the above type , a liquid carrier such as a fatty oil . various other materials may be present as coatings or to modify the physical form of the dosage unit . for instance , tablets can be coated with shellac , sugar , or both . a syrup can contain , in addition to active ingredient , sucrose as a sweetening agent , methyl and propylparabens as preservatives , a dye , and flavoring such as cherry or orange flavor . for oral therapeutic administration , these active compounds can be incorporated with excipients and used in the form of tablets , capsules , elixirs , suspensions , syrups , and the like . such compositions and preparations should contain at least 0 . 1 % of active compound . the percentage of the compound in these compositions can , of course , be varied and can conveniently be between about 2 % to about 60 % of the weight of the unit . the amount of active compound in such therapeutically useful compositions is such that a suitable dosage will be obtained . preferred compositions according to the present invention are prepared so that an oral dosage unit contains between about 1 mg and 800 mg of active compound . the active compounds of the present invention may be orally administered , for example , with an inert diluent , or with an assailable edible carrier , or they can be enclosed in hard or soft shell capsules , or they can be compressed into tablets , or they can be incorporated directly with the food of the diet . the pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions . in all cases , the form should be sterile and should be fluid to the extent that easy syringability exists . it should be stable under the conditions of manufacture and storage and should be preserved against the contaminating action of microorganisms , such as bacteria and fungi . the carrier can be a solvent or dispersion medium containing , for example , water , ethanol , polyol ( e . g ., glycerol , propylene glycol , and liquid polyethylene glycol ), suitable mixtures thereof , and vegetable oils . the compounds or pharmaceutical compositions of the present invention may also be administered in injectable dosages by solution or suspension of these materials in a physiologically acceptable diluent with a pharmaceutical adjuvant , carrier or excipient . such adjuvants , carriers and / or excipients include , but are not limited to , sterile liquids , such as water and oils , with or without the addition of a surfactant and other pharmaceutically and physiologically acceptable components . illustrative oils are those of petroleum , animal , vegetable , or synthetic origin , for example , peanut oil , soybean oil , or mineral oil . in general , water , saline , aqueous dextrose and related sugar solution , and glycols , such as propylene glycol or polyethylene glycol , are preferred liquid carriers , particularly for injectable solutions . the pharmaceutical forms suitable for implantable use include sterile wafers of polycarboxyphenoxypropane - sebacic - acid ( pcpp : sa ) polymers , poly ( d , l - lactic acid ), polyhydroxybutyrate , lysine diisocyanate ( ldi )- glycerol polyurethane , and poly ( d - l lactide - co - glycolide ). in all cases , the form should be sterile and should be a wafer or disc of suitable dimensions for surgical implantation in the brain . the polymers should be stable under the conditions of manufacture and storage and should be preserved against the contaminating action of microorganisms , such as bacteria and fungi . the wafers should be biodegradable ranging from 24 hours up to 6 months . in one aspect , the invention provides compounds and compositions , including any aspect described herein , for use in any of the methods of this invention . in one aspect , use of a compound of this invention or a composition comprising the same , will have utility in inhibiting , suppressing , enhancing or stimulating a desired response in a subject , as will be understood by one skilled in the art . in another embodiment , the compositions may further comprise additional active ingredients , whose activity is useful for the particular application for which the compound of this invention is being administered . therapeutic compositions typically must be sterile and stable under the conditions of manufacture and storage . the composition can be formulated as a solution , microemulsion , liposome , or other ordered structure suitable to high drug concentration . the carrier can be a solvent or dispersion medium containing , for example , water , ethanol , polyol ( for example , glycerol , propylene glycol , and liquid polyethylene glycol , and the like ), and suitable mixtures thereof . the proper fluidity can be maintained , for example , by the use of a coating such as lecithin , by the maintenance of the required particle size in the case of dispersion and by the use of surfactants . in many cases , it will be preferable to include isotonic agents , for example , sugars , polyalcohols such as mannitol , sorbitol , or sodium chloride in the composition . prolonged absorption of the injectable compositions can be brought about by including in the composition an agent which delays absorption , for example , monostearate salts and gelatin . moreover , the modulators can be administered in a time release formulation , for example in a composition which includes a slow release polymer . the active compounds can be prepared with carriers that will protect the compound against rapid release , such as a controlled release formulation , including implants and microencapsulated delivery systems . biodegradable , biocompatible polymers can be used , such as ethylene vinyl acetate , polyanhydrides , polyglycolic acid , collagen , polyorthoesters , polylactic acid and polylactic , polyglycolic copolymers ( plg ). many methods for the preparation of such formulations are patented or generally known to those skilled in the art . the mode of administration may be oral , for intestinal delivery ; intranasal , for nasal delivery ; and intravenous for delivery through the blood - brain barrier . other modes of administration as are known in the art may also be used , including , but not limited to intrathecal , intramuscular , intrabronchial , intrarectal , intraocular , and intravaginal delivery . the modulator compounds can be administered as oral dosage compositions for small intestinal delivery . such oral dosage compositions for small intestinal delivery are well - known in the art , and generally comprise gastroresistent tablets or capsules ( remington &# 39 ; s pharmaceutical sciences , 16th ed ., eds . osol , mack publishing co ., chapter 89 ( 1980 ); digenis et al , j . pharm . sci ., 83 : 915 - 921 ( 1994 ); vantini et al , clinica terapeutica , 145 : 445 - 451 ( 1993 ); yoshitomi et al , chem . pharm . bull ., 40 : 1902 - 1905 ( 1992 ); thoma et al , pharmazie , 46 : 331 - 336 ( 1991 ); morishita et al , drug design and delivery , 7 : 309 - 319 ( 1991 ); and lin et al , pharmaceutical res ., 8 : 919 - 924 ( 1991 )); each of which is incorporated by reference herein in its entirety ). tablets are made gastroresistent by the addition of compounds such as cellulose acetate phthalate or cellulose acetate terephthalate . capsules are solid dosage forms in which the tight junction modulator compound is enclosed in either a hard or soft , soluble container or shell of gelatin . the gelatin used in the manufacture of capsules is obtained from collagenous material by hydrolysis . there are two types of gelatin . type a , derived from pork skins by acid processing , and type b , obtained from bones and animal skins by alkaline processing . the use of hard gelatin capsules permit a choice in prescribing a tight junction modulator compound or a combination thereof at the exact dosage level considered best for the individual subject . the hard gelatin capsule consists of two sections , one slipping over the other , thus completely surrounding the tight junction modulator compound . these capsules are filled by introducing the modulator compound , or gastroresistent beads containing the modulator compound , into the longer end of the capsule , and then slipping on the cap . hard gelatin capsules are made largely from gelatin , fd & amp ; c colorants , and sometimes an opacifying agent , such as titanium dioxide . the usp permits the gelatin for this purpose to contain 0 . 15 % ( w / v ) sulfur dioxide to prevent decomposition during manufacture . in the context of the present invention , oral dosage compositions for small intestinal delivery also include liquid compositions which contain aqueous buffering agents that prevent the modulator compound from being significantly inactivated by gastric fluids in the stomach , thereby allowing the modulator compound to reach the small intestines in an active form . examples of such aqueous buffering agents which can be employed in the present invention include bicarbonate buffer ( ph 5 . 5 to 8 . 7 , preferably about ph 7 . 4 ). when the oral dosage composition is a liquid composition , it is preferable that the composition be prepared just prior to administration so as to minimize stability problems . in this case , the liquid composition can be prepared by dissolving lyophilized tight junction modulator compound in the aqueous buffering agent . oral dosage compositions for small intestinal delivery also include liquid compositions which may optionally contain aqueous buffering agents that prevent the therapeutic agent and tight junction modulator compound from being significantly inactivated by gastric fluids in the stomach , thereby allowing the biologically active ingredient and tight junction modulator compound to reach the small intestines in an active form . examples of such aqueous buffering agents which can be employed in the present invention include bicarbonate buffer ( ph 5 . 5 to 8 . 7 , preferably about ph 7 . 4 ). when the oral dosage composition is a liquid composition , it is preferable that the composition be prepared just prior to administration so as to minimize stability problems . in this case , the liquid composition can be prepared by dissolving lyophilized therapeutic agent and tight junction modulator compound in the aqueous buffering agent . sterile injectable solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above , as required , followed by filtered sterilization . generally , dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other ingredients from those enumerated above . for sterile powders used in the preparation of sterile injectable solutions , the preferred methods of preparation are vacuum drying and freeze - drying which yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile - filtered solution thereof . a “ nasal ” delivery composition differs from an “ intestinal ” delivery composition in that the latter must have gastroresistent properties in order to prevent the acidic degradation of the active agents in the stomach , whereas the former generally comprises water - soluble polymers with a diameter of about 50 11 m in order to reduce the mucociliary clearance , and to achieve a reproducible bioavailability of the nasally administered agents . an “ intravenous ” delivery composition differs from both the “ nasal ” and “ intestinal ” delivery compositions in that there is no need for gastroresistance or water - soluble polymers in the “ intravenous ” delivery composition . nasal dosage compositions for nasal delivery are well - known in the art . such nasal dosage compositions generally comprise water - soluble polymers that have been used extensively to prepare pharmaceutical dosage forms ( martin et al , in : physical chemical principles of 20 pharmaceutical sciences , 3rd ed ., pages 592 - 638 ( 1983 )) that can serve as carriers for peptides for nasal administration ( davis , in : delivery systems for peptide drugs , 125 : 1 - 21 ( 1986 )). the nasal absorption of pap tides embedded in polymer matrices has been shown to be enhanced through retardation of nasal mucociliary clearance ( illum et al , int . j . pharm ., 46 : 261 - 265 ( 1988é ). other possible enhancement mechanisms include an increased concentration gradient or 25 decreased diffusion path for peptides absorption ( ting et al , pharm . res ., 9 : 1330 - 1335 ( 1992 ). however , reduction in mucociliary clearance rate has been predicted to be a good approach toward achievement or reproducible bioavailability of nasally administered systemic drugs ( gonda et al , pharm . res ., 7 : 69 - 75 ( 1990 )). microparticles with a diameter of about 50 pm are expected to deposit in the nasal cavity ( bjork et al , int . j . pharm ., 62 : 187 - 192 ( 1990 ); and lllum et al , int . j . pharm ., 39 : 189 - 199 ( 1987 ), while microparticles with a diameter under 10 pm can escape the filtering system of the nose and deposit in the lower airways . microparticles larger than 200 pm in diameter will not be retained in the nose after nasal administration ( lewis et al , proc . int . symp . control rei . bioact . mater ., 17 : 280 - 290 ( 1990 )). the particular water - soluble polymer employed is not critical to the present invention , and can be selected from any of the well - known water - soluble polymers employed for nasal dosage forms . a typical example of a water - soluble polymer useful for nasal delivery is polyvinyl alcohol ( pva ). this material is a swellable hydrophilic polymer whose physical properties depend on the molecular weight , degree of hydrolysis , cross - linking density , and crystallinity ( peppas et al , in : hydrogels in medicine and pharmacy , 3 : 109 - 131 ( 1987 ). pya can be used in the coating of dispersed materials through phase separation , spray - drying , spray - embedding , and spray - densation ( ting et al , supra ). a “ skin ” delivery composition comprising a modulator compound of the invention may include in addition a therapeutic or immunogenic agent , fragrance , creams , ointments , colorings , and other compounds so long as the added component does not deleteriously affect transdermal delivery of the therapeutic or immunogenic agent . conventional pharmaceutically acceptable emulsifiers , surfactants , suspending agents , antioxidants , osmotic enhancers , extenders , diluents and preservatives may also be added . water soluble polymers can also be used as carriers . the particular therapeutic or immunogenic agent employed is not critical to the present invention , and can be , e . g ., any drug compound , biologically active peptide , vaccine , or any other moiety otherwise not absorbed through the transcellular pathway , regardless of size or charge . the amount of active compound in the composition may vary according to factors such as the disease state , age , sex , and weight of the individual . dosage regimens may be adjusted to provide the optimum therapeutic response . for example , a single bolus may be administered , several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation . it is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage . dosage unit form as used herein refers to physically 35 discrete units suited as unitary dosages for the mammalian subjects to be treated ; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier . the specification for the dosage unit forms of the invention are dictated by and directly dependent on ( a ) the unique characteristics of the active compound and the particular therapeutic effect to be achieved , and ( b ) the limitations inherent in the art of compounding such an active compound for the treatment of sensitivity in individuals . as used herein “ pharmaceutically - acceptable carrier ” includes any and all solvents , dispersion media , coatings , antibacterial and antifungal agents , isotonic and absorption delaying agents , and the like that are physiologically compatible . in one embodiment , the carrier is 10 suitable for parenteral administration . a carrier may be suitable for administration into the central nervous system ( e . g ., intraspinally or intracerebrally ). alternatively , the carrier can be suitable for intravenous , intraperitoneal or intramuscular administration . in another embodiment , the carrier is suitable for oral administration . pharmaceutically - acceptable carriers include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion . the use of such media and agents for pharmaceutically active substances is well known in the art . except insofar as any conventional media or agent is incompatible with the active compound , use thereof in the pharmaceutical compositions of the invention is contemplated . supplementary active compounds can also be incorporated into the compositions . while specific aspects of the invention have been described in detail , it will be appreciated by those skilled in the art that various modifications and alternatives to those details could be developed in light of the overall teachings of the disclosure . accordingly , the particular arrangements disclosed are meant to be illustrative only , and not limiting as to the scope of the invention , which is to be given the full breadth of the appended claims , and any equivalent , thereof . the foregoing discussion may be better understood in connection with the following representative examples which are presented for purposes of illustrating the principle methods and compositions of the invention , and not by way of limitation . various other examples will be apparent to the person skilled in the art after reading the present disclosure without departing from the spirit and scope of the invention . it is intended that all such other examples be included within the scope of the appended claims . all parts are by weight ( e . g ., % w / w ), and temperatures are in degrees centigrade (° c . ), unless otherwise indicated . melting points were determined with a hoover melting point apparatus and are uncorrected . infrared ( ir ) spectra for the compounds were recorded in kbr discs on a mattson satellite ftir in cm − 1 . 1 h and 13 c spectra were recorded in dmso - d 6 on a bruker avance iii dpx 300 mhz instrument . 19 f spectra were recorded in dmso d 6 on a bruker avance iii 600 ( 564 . 6 mhz ). chemical shifts were expressed in parts per million ( δ ) with tetramethylsilane as internal standard . mass spectrometry was performed on a thermo scientific ltq - ft at the university of cincinnati mass spectrometry facility . the purity of the compounds was monitored by hplc using a waters 2695 separation module and a 2487 dual λ absorbance detector with a novapak c18 4 μm 3 . 9 × 150 mm column . the mobile phases consisted of acetonitrile / h 2 o using a 30 minute gradient . all compounds were ≧ 95 %. microanalysis was performed by atlantic microlab inc ., and all compounds were found to be ± 0 . 4 %. all reagents were from sigma - aldrich . logs , logp , log bbb , human intestinal absorption , p - glycoprotein category , cyp 2c9 pki , herg pic50 , cyp 2d6 affinity category , oral cns score , iv cns score , mw , flexibility , and total polar surface area were calculated using stardrop 5 . 1 . 1 release build 178 . scheme 1 illustrates the synthetic reactions used to summarize these reactions . table 2 is a non - limiting list of aryl functional groups that can be incorporated as “ ar 1 ” or “ ar 2 ” from formulae ia , ib , and iia . tables 3 and 4 illustrate the structures , names , and numbers of a variety of key compounds disclosed in this application . potassium cyanide ( 0 . 91 g ) was added to sodium carbonate ( 1 . 7 g ) in deionized water ( 30 ml ) in a 3 - neck glass round flask and placed in an ice bath . the system was repeatedly purged using a vacuum pump and nitrogen gas . glyoxal ( 3 . 72 g ) was then added to the system without the introduction of o 2 and the reactants were allowed to dissolve with stirring . in a stoppered tube , the appropriate arylaldehyde ( 7 . 11 mmoles ) was added to 1 , 4 - dioxane ( 5 ml ), purged , and then added drop - wise to the system . the system was then removed from the ice bath and allowed to stir at room temperature for 1 hour . after 1 hour , acetic acid ( 5 ml ) was added drop - wise until gas bubbles were no longer visible from the addition of acetic acid , or until the solution was at a ph of less than 6 . the solution was vacuum filtered and washed with ice cold water ( 5 ml ), methanol ( 5 ml ) and ether ( 5 ml ) and then was allowed to air dry . crude material was recrystallized with methanol , collected by vacuum filtration and rinsed with diethyl ether and dried under vacuum . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of methanesulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours , followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of ethanesulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours , followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirring at room temperature for 30 minutes , and 1 eq of n - propanesulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours , followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether , the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of i - propanesulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours , followed by the addition of a saturated ammonium chloride . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of n - butanesulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours , followed by the addition of a saturated ammonium chloride . the mixture is extracted 3 times with 30 ml of diethyl ether , the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirring at room temperature for 30 minutes , and 1 eq of i - butanesulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours , followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one was stirred in 50 ml of dry thf under argon gas for 16 hours with 4 . 7 g k 2 co 3 and 4 . 25 ml of benzenesulfonyl chloride . the reaction was filtered , and the filtrate was acidified with 24 ml 1n hcl , and extracted 5 times with 20 ml of diethyl ether . the combined ether extracts were washed with brine and dried with na 2 so 4 . after filtration , 100 ml of hexanes was added to the solution , resulting in a precipitate , which was collected using vacuum filtration and recrystallized with meoh . yield = 17 %. mp 190 - 195 ° c . ; ftir 3099 , 1630 ; 1 h nmr ( 300 mhz , dmso - d 6 , ppm ) δ 8 . 54 ( s , 2h ), 7 . 93 ( d , j = 8 . 1 hz , 2h ), 7 . 74 ( t , j = 7 . 2 hz , 1h ), 7 . 57 ( t , j = 8 . 1 hz , 2h ), 7 . 48 ( d , j = 8 . 7 hz , 2h ), 7 . 18 ( d , j = 8 . 4 hz , 2h ), 5 . 54 ( s , 1h ). 13 c nmr ( 75 mhz , dmso - d 6 , ppm ) δ 181 . 2 , 173 . 4 , 135 . 2 , 135 . 0 , 134 . 2 , 133 . 9 , 129 . 7 , 129 . 2 , 129 . 1 , 129 . 0 , 106 . 6 , 82 . 9 . elemental analysis calc : c 52 . 54 , h 3 . 31 , n 3 . 83 , cl 9 . 69 ; found : c 52 . 50 , h 3 . 33 , n 3 . 79 , cl 9 . 84 ; c 16 h 12 clno 5 s ; hplc retention time : 32 . 2 min . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 2 - chlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 3 - chlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether , the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . 2 . 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one was stirred in 100 ml of dry thf . 1 . 31 ml of tea was added , and 30 minutes later 2 . 11 g of phenylmethylsulfonylchloride was added to the reaction . the reaction was stirred for 24 hours . the reaction was filtered , and the filtrate was acidified with 24 ml 1n hcl , and extracted 5 times with 20 ml of diethyl ether . the combined ether extracts were washed with brine , and dried with na 2 so 4 . after filtration , the solution was concentrated under reduced pressure , and the resulting solid was recrystallized from methanol . yield = 27 %. ftir 2957 , 1636 ; 1 h nmr ( 300 mhz , dmso - d 6 , ppm ) δ 8 . 79 ( s , 2h ), 7 . 60 - 7 . 49 ( m , 4h ), 7 . 43 - 7 . 35 ( m , 5h ), 5 . 80 ( s , 1h ), 4 . 97 ( d , j = 14 . 1 , 1h ), 4 . 90 ( d , j = 14 . 1 , 1h ). 13 c nmr ( 75 mhz , dmso - d 6 , ppm ) δ 181 . 8 , 173 . 9 , 134 . 3 , 134 . 1 , 131 . 5 , 129 . 3 , 129 . 1 , 129 . 1 , 129 . 0 , 128 . 9 , 107 . 9 , 83 . 1 , 57 . 8 . elemental analysis calc : c 53 . 76 , h 3 . 72 , n 3 . 69 ; found : c 53 . 90 , h 3 . 68 , n 3 . 70 ; c 17 h 14 clno 5 s ; hplc retention time : 32 . 2 min . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 2 , 3 - dichlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 2 , 4 - dichlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours , followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 2 , 5 - dichlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 3 - carboxymethylphenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 3 , 4 - dichlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 3 , 5 - dichlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 2 - fluorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours , followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 3 - fluorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - fluorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours , followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 2 , 3 - difluorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours , followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 2 , 4 - difluorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 2 , 5 - difluorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 2 , 6 - difluorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 3 , 4 - difluorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 3 , 5 - difluorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - trifluoromethylphenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirring at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - nitrilephenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 3 - nitrilephenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of 4 - fluorophenylmethylsulfonyl chloride in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one in dry thf under argon is added triethylamine in dry thf . the reaction is stirred at room temperature for 30 minutes , and 1 eq of phenylchloroformate in dry thf is added dropwise . the reaction is stirred for 16 hours followed by the addition of a saturated ammonium chloride solution . the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . to a stirring solution of 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one and sodium carbonate ( 1 . 7 g ) in deionized water ( 30 ml ) is added phenylisothiocyanate . the reaction is stirred at room temperature for 24 hours . a saturated ammonium chloride solution is added and the mixture is extracted 3 times with 30 ml of diethyl ether . the combined ether extracts are washed with water and brine , dried over sodium sulfate , filtered , and concentrated under reduced pressure . the solid is recrystallized with methanol . potassium cyanide ( 0 . 91 g ) was added to sodium carbonate ( 1 . 7 g ) in deionized water ( 30 ml ) in a 3 - neck glass round flask and placed in an ice bath . the system was repeatedly purged using a vacuum pump and nitrogen gas . glyoxal ( 3 . 72 g ) was then added to the system without the introduction of o 2 and the reactants were allowed to dissolve with stirring . in a stoppered tube , the appropriate arylaldehyde ( 7 . 11 mmoles ) was added to 1 , 4 - dioxane ( 5 ml ), purged , and then added drop - wise to the system . the system was then removed from the ice bath and allowed to stir at room temperature for 1 hour . after 1 hour , acetic acid ( 5 ml ) was added drop - wise until gas bubbles were no longer visible from the addition of acetic acid , or until the solution was at a ph of less than 6 . the solution was vacuum filtered and washed with ice cold water ( 5 ml ), methanol ( 5 ml ) and ether ( 5 ml ) and then was allowed to air dry . crude material was recrystallized with methanol , collected by vacuum filtration and rinsed with diethyl ether and dried under vacuum . yield = 70 %. mp 221 - 2 ° c . ; ftir 3079 , 1638 ; 1 h nmr ( 300 mhz , dmso - d 6 , ppm ) δ 7 . 82 ( s , 2h ), 7 . 46 ( d , j = 8 . 7 hz , 2h ), 7 . 30 ( s , 1h ), 7 . 29 ( d , j = 8 . 7 hz , 2h ), 5 . 43 ( s , 1h ). 13 c nmr ( 75 mhz , dmso - d 6 , ppm ) δ 182 . 6 , 173 . 1 , 135 . 7 , 133 . 2 , 128 . 9 , 128 . 6 , 111 . 7 , 82 . 2 . hrms calc : 248 . 00849 , found : 248 . 00852 mna + ═ c 10 h 8 no 3 clna + ; elemental analysis calc : c 53 . 23 , h 3 . 57 , n 6 . 21 , cl 15 . 71 ; found : c 53 . 35 , h 3 . 61 , n 6 . 24 , cl 15 . 83 c 10 h 8 clno 3 ; hplc retention time : 16 . 7 min . yield = 84 %. mp 160 - 3 ° c . ; ftir 3079 , 1638 ; 1 h nmr ( dmso d 6 ) 5 . 60 ( 1h , s ), 7 . 22 - 7 . 78 ( 4h , m ); hrms calc : 210 . 05610 , found : 210 . 05609 mh + = c 10 h 9 fno 3 + ; elemental analysis calc : c 57 . 42 , h 3 . 85 , n 6 . 70 , f 9 . 08 ; found : c 57 . 50 , h 3 . 92 , n 6 . 61 , f 8 . 99 c 10 h 8 fno 3 ; hplc retention time : 11 . 42 min . yield = 60 %. mp 168 ° c . ; ftir 3356 , 3129 ; 1 h nmr ( dmso d 6 ) 5 . 44 ( 1h , s ), 7 . 05 - 7 . 27 ( 4h , m ); hrms calc : 210 . 05610 , found : 210 . 05609 mh + = c 10 h 9 f no 3 +; elemental analysis calc : c 57 . 42 , h 3 . 85 , n 6 . 70 , f 9 . 08 ; found : c 57 . 41 , h 3 . 87 , n 6 . 61 , f 8 . 97 c 10 h 8 fno 3 ; hplc retention time : 12 . 63 min . yield = 68 %. mp 160 ° c . ; ftir 3351 , 3138 ; 1 h nmr ( dmso d 6 ) 5 . 41 ( 1h , s ), 7 . 22 - 7 . 78 ( 4h , m ); hrms calc : 210 . 05610 , found : 210 . 05609 mh + = c 10 h 9 fno 3 + ; elemental analysis calc : c 57 . 42 , h 3 . 85 , n 6 . 70 , f 9 . 08 ; found : c 57 . 42 , h 3 . 97 , n 6 . 66 , f 8 . 90 c 10 h 8 fno 3 ; hplc retention time : 11 . 88 min . yield = 76 %. mp 193 ° c . ; ftir 3391 , 3277 , 1539 ; 1 h nmr ( dmso d 6 ) 5 . 68 ( 1h , s ), 7 . 05 - 7 . 85 ( 3h , m ); hrms calc : 228 . 04668 , found : 228 . 04669 mh + = c 10 h 8 f 2 no 3 + ; elemental analysis calc : c 52 . 87 , h 3 . 11 , n 6 . 17 , f 16 . 73 ; found : c 53 . 10 , h 3 . 11 , n 6 . 17 , f 16 . 57 c 10 h 7 f 2 no 3 ; hplc retention time : 13 . 02 min . yield = 67 %. mp 182 - 3 ° c . ; ftir 3252 , 1608 ; 1 h nmr ( dmso d 6 ) 5 . 59 ( 1h , s ), 7 . 13 - 7 . 79 ( 3h , m ); hrms calc : 228 . 04668 , found : 228 . 04671 mh + = c 10 h 8 f 2 no 3 + ; elemental analysis calc : c 52 . 87 , h 3 . 11 , n 6 . 17 , f 16 . 73 ; found : c 52 . 84 , h 3 . 00 , n 6 . 16 , f 16 . 59 c 10 h 7 f 2 no 3 ; hplc retention time : 12 . 63 min . yield = 80 %. mp 196 ° c . ; ftir 3391 , 3267 ; 1 h nmr ( dmso d 6 ) 5 . 61 ( 1h , s ), 7 . 03 - 7 . 84 ( 3h , m ); hrms calc : 228 . 04668 , found : 228 . 04670 mh + = c 10 h 8 f 2 no 3 + ; elemental analysis calc : c 52 . 87 , h 3 . 11 , n 6 . 17 , f 16 . 73 ; found : c 52 . 79 , h 3 . 11 , n 6 . 15 , f 16 . 57 c 10 h 7 f 2 no 3 ; hplc retention time : 12 . 09 min . yield = 70 %. mp 159 - 60 ° c . ; ftir 3535 , 3406 ; 1 h nmr ( dmso d 6 ) 5 . 68 ( 1h , s ), 7 . 14 - 7 . 73 ( 3h , m ); hrms calc : 228 . 04668 , found : 228 . 04670 mh + = c 10 h 8 f 2 no 3 + ; elemental analysis calc : c 52 . 87 , h 3 . 11 , n 6 . 17 , f 16 . 73 ; found : c 52 . 62 , h 3 . 10 , n 5 . 94 , f 16 . 57 c 10 h 7 f 2 no 3 ; hplc retention time : 11 . 30 min . yield = 76 %. mp 190 - 4 ° c . ; ftir 3322 , 3124 ; 1 h nmr ( dmso d 6 ) 5 . 44 ( 1h , s ), 7 . 13 - 7 . 85 ( 3h , m ); hrms calc : 228 . 04668 , found : 228 . 04670 mh + = c 10 h 8 f 2 no 3 + ; elemental analysis calc : c 52 . 87 , h 3 . 11 , n 6 . 17 , f 16 . 73 ; found : c 53 . 13 , h 3 . 16 , n 6 . 15 , f 16 . 61 c 10 h 7 f 2 no 3 ; hplc retention time : 13 . 79 min . yield = 58 %. mp 190 - 1 ° c . ; ftir 3346 , 3143 ; 1 h nmr ( dmso d 6 ) 5 . 68 ( 1h , s ), 7 . 05 - 7 . 85 ( 3h , m ); hrms calc : 228 . 04668 , found : 228 . 04670 mh + = c 10 h 8 f 2 no 3 + ; elemental analysis calc : c 52 . 87 , h 3 . 11 , n 6 . 17 , f 16 . 73 ; found : c 52 . 88 , h 3 . 06 , n 6 . 15 , f 16 . 70 c 10 h 7 f 2 no 3 ; hplc retention time : 14 . 00 min . 1 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one was stirred in acetic anhydride under nitrogen gas for 16 hours . the reaction was cooled to − 78 ° c ., and lyophilized . the dried mass was recrystallized from meoh . yield = 44 %. mp 221 - 2 ° c . ; ftir 3030 , 1628 ; 1 h nmr ( 300 mhz , dmso - d 6 , ppm ) δ 8 . 30 ( s , 2h ), 7 . 47 ( d , j = 8 . 7 hz , 2h ), 7 . 35 ( d , j = 8 . 7 hz , 2h ), 5 . 63 ( s , 1h ), 2 . 14 ( s , 3h ). 13 c nmr ( 75 mhz , dmso - d 6 , ppm ) δ 182 . 4 , 173 . 0 , 168 . 9 , 134 . 8 , 133 . 8 , 129 . 1 , 129 . 1 , 106 . 9 , 83 . 2 , 20 . 7 . elemental analysis calc : c 53 . 85 , h 3 . 77 , n 5 . 23 ; found : c 53 . 76 , h 3 . 90 , n 5 . 24 ; c 12 h 10 clno 4 ; hplc retention time : 19 . 7 min . 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one was stirred in 50 ml of dry thf under nitrogen gas for 16 hours with 4 . 7 g k 2 co 3 and 4 . 25 ml of benzenesulfonyl chloride . the reaction was filtered , and the filtrate was acidified with 24 ml 1n hcl , and extracted 5 times with 20 ml of diethyl ether . the combined ether extracts were washed with brine , and dried with na 2 so 4 . after filtration , 100 ml of hexanes was added to the solution , resulting in a precipitate , which was collected using vacuum filtration and recrystallized with meoh . yield = 17 %. mp 190 - 195 ° c . ; ftir 3034 , 1630 ; 1 h nmr ( 300 mhz , dmso - d 6 , ppm ) δ 8 . 54 ( s , 2h ), 7 . 93 ( d , j = 8 . 1 hz , 2h ), 7 . 74 ( t , j = 7 . 2 hz , 1h ), 7 . 57 ( t , j = 8 . 1 hz , 2h ), 7 . 48 ( d , j = 8 . 7 hz , 2h ), 7 . 18 ( d , j = 8 . 4 hz , 2h ), 5 . 54 ( s , 1h ). 13 c nmr ( 75 mhz , dmso - d 6 , ppm ) δ 181 . 2 , 173 . 4 , 135 . 2 , 135 . 0 , 134 . 2 , 133 . 9 , 129 . 7 , 129 . 2 , 129 . 1 , 129 . 0 , 106 . 6 , 82 . 9 . elemental analysis calc : c 52 . 54 , h 3 . 31 , n 3 . 83 , cl 9 . 69 ; found : c 52 . 50 , h 3 . 33 , n 3 . 79 , cl 9 . 84 ; c 16 h 12 clno 5 s ; hplc retention time : 32 . 2 min . 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one was stirred in 50 ml of dry thf under nitrogen gas for 16 hours with 4 . 6 g k 2 co 3 and 1 . 5 ml of methanesulfonyl chloride . the reaction was filtered , and the filtrate was acidified with 24 ml 1n hcl , and extracted 5 times with 20 ml of diethyl ether . the combined ether extracts were washed with brine , and dried with na 2 so 4 . after filtration , 100 ml of hexanes was added to the solution , resulting in a precipitate , which was collected using vacuum filtration and recrystallized with meoh . the material was further purified by column chromatography . yield = 18 %. mp 175 ° c . ; ftir 3169 , 1616 ; 1 h nmr ( 300 mhz , dmso - d 6 , ppm ) δ 8 . 41 ( s , 1h ), 8 . 09 ( s , 1h ), 7 . 96 ( d , j = 8 . 4 hz , 2h ), 7 . 85 ( s , 1h ), 7 . 59 ( d , j = 8 . 4 hz , 2h ), 3 . 55 ( s , 3h ). 13 c nmr ( 75 mhz , dmso - d 6 , ppm ) δ 185 . 7 , 165 . 1 , 140 . 7 , 136 . 9 , 135 . 9 , 133 . 5 , 130 . 0 , 129 . 7 , 40 . 7 . elemental analysis calc : c 43 . 50 , h 3 . 32 , cl 11 . 67 , n 4 . 61 ; found : c 43 . 66 , h 3 . 40 , cl 11 . 54 , n 4 . 55 ; c 11 h 10 clno 5 s ; hplc retention time : 25 . 2 min . 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - chlorophenyl )- furan - 3 - one was stirred in 50 ml of dry thf under nitrogen gas for 16 hours with 4 . 6 g k 2 co 3 and 2 . 1 ml of ethanesulfonyl chloride . the reaction was filtered , and the filtrate was acidified with 24 ml 1n hcl , and extracted 5 times with 20 ml of diethyl ether . the combined ether extracts were washed with brine , and dried with na 2 so 4 . after filtration , 100 ml of hexanes was added to the solution , resulting in a precipitate , which was collected using vacuum filtration and recrystallized with ethyl acetate . yield = 21 %. mp 183 - 185 ° c . ; ftir 3181 , 1616 ; 1 h nmr ( 300 mhz , dmso - d 6 , ppm ) δ 8 . 41 ( s , 1h ), 8 . 08 ( s , 1h ), 7 . 95 ( d , j = 9 . 3 hz , 2h ), 7 . 85 ( s , 1h ), 7 . 59 ( d , j = 9 . 0 hz , 2h ), 3 . 68 ( q , j = 7 . 2 hz , 2h ), 1 . 40 ( t , j = 6 . 9 hz , 3h ). 13 c nmr ( 75 mhz , dmso - d6 , ppm ) δ 185 . 8 , 165 . 1 , 140 . 7 , 136 . 8 , 136 . 1 , 133 . 5 , 130 . 0 , 129 . 7 , 48 . 0 , 8 . 6 . elemental analysis calc : c 45 . 36 , h 3 . 81 , cl 11 . 16 , n 4 . 41 ; found : c 45 . 42 , h 3 . 85 , cl 11 . 06 , n 4 . 37 ; c 12 h 12 clno 5 s ; hplc retention time : 29 . 9 min . 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - fluorophenyl )- furan - 3 - one is stirring in 50 ml dry thf under dry nitrogen . the reaction is cooling in an ice bath , and 1 equivalent of triethylamine is added dropwise . the reaction is warmed to room temperature , chlorotrimethylsilane is added dropwise and the reaction is refluxing gently with a water bath for 30 minutes . 1 equivalent of benzene sulfonyl chloride is added dropwise . 1 equivalent of tea is added dropwise , and the reaction is gently refluxing with a water bath for 1 hour . the reaction is cooling to room temperature , and 1 equivalent of tetrabutylammonium fluoride is added and stirs for 30 minutes . a saturated ammonium sulfate solution is added to quench the reaction , and the reaction is extracted 3 times with 20 ml diethyl ether . the combined ether extracts are washed with water and brine , the ether is dried with sodium sulfate , is filtered , and evaporates to yield a solid which is recrystallized with methanol . 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - fluorophenyl )- furan - 3 - one is stirring in 50 ml dry thf under dry nitrogen . the reaction is cooling in an ice bath , and 1 equivalent of triethylamine is added dropwise . the reaction is warmed to room temperature , chlorotrimethylsilane is added dropwise and the reaction is refluxing gently with a water bath for 30 minutes . 1 equivalent of acetyl chloride is added dropwise . 1 equivalent of tea is added dropwise , and the reaction is gently refluxing with a water bath for 1 hour . the reaction is cooling to room temperature , and 1 equivalent of tetrabutylammonium fluoride is added and stirs for 30 minutes . a saturated ammonium sulfate solution is added to quench the reaction , and the reaction is extracted 3 times with 20 ml diethyl ether . the combined ether extracts are washed with water and brine , the ether is dried with sodium sulfate , is filtered , and evaporates to yield a solid which is recrystallized with methanol . 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - fluorophenyl )- furan - 3 - one is stirring in 50 ml dry thf under dry nitrogen . the reaction is cooling in an ice bath , and 1 equivalent of triethylamine is added dropwise . the reaction is warmed to room temperature , chlorotrimethylsilane is added dropwise and the reaction is refluxing gently with a water bath for 30 minutes . 1 equivalent of benzoyl chloride is added dropwise . 1 equivalent of tea is added dropwise , and the reaction is gently refluxing with a water bath for 1 hour . the reaction is cooling to room temperature , and 1 equivalent of tetrabutylammonium fluoride is added and stirs for 30 minutes . a saturated ammonium sulfate solution is added to quench the reaction , and the reaction is extracted 3 times with 20 ml diethyl ether . the combined ether extracts are washed with water and brine , the ether is dried with sodium sulfate , is filtered , and evaporates to yield a solid which is recrystallized with methanol . 5 g of 5 - amino - 4 - hydroxy - 2 -( 4 - fluorophenyl )- furan - 3 - one is stirring in 50 ml pyridine under dry nitrogen . 1 equivalent of succinic anhydride is added , and the reaction is refluxing gently with a water bath for 1 hour . a saturated ammonium sulfate solution is added to quench the reaction , and the reaction is extracted 3 times with 20 ml diethyl ether . the combined ether extracts are washed with a saturated bicarbonate solution and brine , the ether is dried with sodium sulfate , is filtered , and evaporates to yield a solid which is recrystallized with methanol . while this invention has been particularly shown and described with references to preferred embodiments thereof , it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims . the patent and scientific literature referred to herein establishes the knowledge that is available to those with skill in the art . all united states patents and published or unpublished united states patent applications cited herein are incorporated by reference . all published foreign patents and patent applications cited herein are hereby incorporated by reference . all other published references , documents , manuscripts and scientific literature cited herein are hereby incorporated by reference . 1 . u . s . pat . no . 6 , 797 , 696 ; issued sep . 28 , 2004 “ diagnosis and treatment of malignant neoplasms ” by wands ; jack r . 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