Novel antiproliferative factor and methods of use

Abstract

A novel antiproliferative factor comprising a glycopeptide is disclosed. In specific embodiments, the novel antiproliferative factor is associated with the bladder. Compositions, diagnostic kits and reagents, and methods of using the compounds for identifying and/or treating interstitial cystitis and cancer are disclosed.

1 . An isolated or synthesized composition comprising a urinary bladder antiproliferative factor having one or more sugar moieties, wherein at least one sugar moiety is linked to a hydrophobic moiety.  
     
     
         2 . The composition of  claim 1 , further defined as a urinary bladder epithelial cell antiproliferative factor.  
     
     
         3 . The composition of  claim 1 , wherein the hydrophobic moiety comprises one of the following: 
 a peptide moiety; or    a lipid moiety.    
     
     
         4 . The method of  claim 3 , wherein the peptide moiety is linear, cyclical, or branched.  
     
     
         5 . The method of  claim 3 , wherein the peptide has homology to at least part of a frizzled polypeptide.  
     
     
         6 . The method of  claim 5 , wherein the peptide is further defined as having homology to at least part of a transmembrane domain of frizzled  8 .  
     
     
         7 . The composition of  claim 3 , further defined as comprising: 
 about one to about six sugar moieties; and    a peptide moiety of about two to about fifteen amino acid residues, wherein one of the residues is a linking amino acid, and wherein the linking amino acid comprises a heteroatom covalently linked to one of the sugar moieties.    
     
     
         8 . The composition of  claim 7 , wherein the linking amino acid is polar.  
     
     
         9 . The composition of  claim 7 , wherein the linking amino acid is a serine, threonine, or cysteine.  
     
     
         10 . The composition of  claim 7 , further defined as comprising two sugar residues and nine amino acids, wherein said linking amino acid is a serine, a threonine or a cysteine.  
     
     
         11 . The composition of  claim 7 , further defined as comprising three sugar residues and nine amino acids, wherein said linking amino acid is a serine, a threonine, or a cysteine.  
     
     
         12 . The composition of  claim 1 , wherein said sugar moiety comprises a naturally occurring sugar, a synthetic sugar, a derivative of a naturally occurring sugar, or a derivative of a synthetic sugar.  
     
     
         13 . The composition of  claim 3 , wherein the linkage between a sugar and a peptide is in the alpha or beta configuration.  
     
     
         14 . The composition of  claim 3 , wherein the peptide moiety is further defined as comprising a naturally occurring amino acid, an unnatural amino acid, a derivative of a naturally occurring amino acid, a derivative of an unnatural amino acid, a modified amino acid, a backbone-modifying amino acid, or a mixture thereof.  
     
     
         15 . The composition of  claim 14 , wherein the peptide moiety is further defined as comprising one or more backbone-modifying amino acids that comprise reduced peptide bonds.  
     
     
         16 . The composition of  claim 14 , wherein the modified amino acid is further defined as a methylated amino acid, an acetylated amino acid, a beta amino acid, or an amino acid mimetic.  
     
     
         17 . The composition of  claim 3 , wherein the peptide moiety comprises SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, or functional derivatives thereof.  
     
     
         18 . The composition of  claim 3 , wherein the lipid moiety comprises a satured fatty acid, an unsaturated fatty acid, an esterified fatty acid, an omega fatty acid, a phosphate-containing fat, or a surfactant having a polar head and a non-polar tail.  
     
     
         19 . The composition of  claim 1 , further defined as comprising a label.  
     
     
         20 . The composition of  claim 19 , wherein the label comprises a fluorescent moiety, a colorimetric moiety, or a radioactive moiety.  
     
     
         21 . The composition of  claim 1 , further defined as comprising: 
 (a) Sialic acid-galactose-Nacetylgalactosamine-threonine-valine-proline-alanine-alanine-valine-valine-valine-alanine;    (b) Sialic acid-galactose-Nacetylglucosamine-threonine-valine-proline-alanine-alanine-valine-valine-valine-alanine; or    (c) Sialic acid-galactose-Nacetylglucosamine-serine-leucine-proline-alanine-alanine-valine-valine-valine-alanine.    
     
     
         22 . The composition of (a) in  claim 21 , further defined as having one or more of the following: 
 the sialic acid is linked to galactose via a 2,3 linkage;    the galactose is linked to the N-acetylgalactosamine via a 1,3 linkage; or    the N-acetyl galactosamine is linked to threonine via an O linkage in an alpha configuration.    
     
     
         23 . The composition of (b) in  claim 21 , further defined as having one or more of the following: 
 the sialic acid is linked to galactose via a 2,3 linkage;    the galactose is linked to the N-acetylglucosamine via a 1,4 linkage; or    the N-acetyl glucosamine is linked to threonine via an O linkage in an alpha configuration.    
     
     
         24 . The composition of (c) in  claim 21 , further defined as having one or more of the following: 
 the sialic acid is linked to galactose via a 2,3 linkage;    the galactose is linked to the N-acetylglucosamine via a 1,4 linkage; or    the N-acetylglucosamine is linked to serine via an O linkage in an alpha configuration.    
     
     
         25 . The composition of  claim 1 , further comprising a delivery vehicle.  
     
     
         26 . The composition of  claim 25 , wherein the delivery vehicle is a liposome, a cell, a conjugated molecule, a nanoparticle, or a mixture thereof.  
     
     
         27 . The composition of  claim 26 , wherein the conjugated molecule comprises an antibody, a peptide, folate, polyethylene glycol, a drug, a liposome, or lactosaminated human albumin.  
     
     
         28 . The method of  claim 26 , wherein the nanoparticle comprises colloidal gold.  
     
     
         29 . The composition of  claim 1  comprised in a pharmaceutically acceptable excipient.  
     
     
         30 . The composition of  claim 1 , wherein the composition reversibly arrests cell proliferation.  
     
     
         31 . The composition of  claim 1 , wherein the composition is further defined as comprising activity for arresting cell cycling in G2.  
     
     
         32 . An isolated peptide selected from the group consisting of SEQ ID NO:1; SEQ ID NO:3; SEQ ID NO:4; SEQ ID NO:5, SEQ ID NO:6, or a functional derivative thereof.  
     
     
         33 . The isolated peptide of  claim 32 , wherein the functional derivative thereof is further defined as comprising a conservative substitution at one or more amino acids of the peptide.  
     
     
         34 . The isolated peptide of  claim 33 , wherein the conservative substitution is further defined as a substitution at serine, threonine, proline, or a combination thereof.  
     
     
         35 . The isolated peptide of  claim 33 , wherein the conservative substitution comprises a hydrophobic conservative substitution.  
     
     
         36 . The isolated peptide of  claim 35 , wherein the hydrophobic conservative substitution is further defined as a substitution at one or more alanines, one or more valines, one or more prolines; or a combination thereof.  
     
     
         37 . An isolated polynucleotide encoding SEQ ID NO:1.  
     
     
         38 . The polynucleotide of  claim 37 , further defined as SEQ ID NO:2.  
     
     
         39 . An isolated polynucleotide encoding SEQ ID NO:3.  
     
     
         40 . An isolated polynucleotide encoding SEQ ID NO:4.  
     
     
         41 . An isolated polynucleotide encoding SEQ ID NO:5.  
     
     
         42 . An isolated polynucleotide encoding SEQ ID NO:6.  
     
     
         43 . A kit, comprising the composition of  claim 1  housed in a suitable container.  
     
     
         44 . A kit comprising a detection means for detecting the composition of  claim 1 , said detection means housed in a suitable container.  
     
     
         45 . A kit for use in diagnosing a bladder disorder in a subject, comprising an antibody composition that binds immunologically to an epitope of a peptide, sugar, or glycopeptide moiety of an urinary bladder antiproliferative factor, said antibody composition housed in a suitable container.  
     
     
         46 . The kit of  claim 45 , wherein said peptide or glycopeptide comprises one or more hydrophobic amino acids.  
     
     
         47 . The kit of  claim 45 , wherein the antibody binds immunologically to an epitope of SEQ ID NO:1 or a glycopeptide derivative thereof.  
     
     
         48 . The kit of  claim 47 , wherein the peptide comprising an epitope of SEQ ID NO:1 is about 2 to 15 amino acids in length.  
     
     
         49 . The kit of  claim 45 , wherein the antibody binds immunologically to an epitope of SEQ ID NO:3 or a glycopeptide derivative thereof.  
     
     
         50 . The kit of  claim 49 , wherein the peptide comprising an epitope of SEQ ID NO:3 is about 2 to 15 amino acids in length.  
     
     
         51 . The kit of  claim 45 , wherein the antibody binds immunologically to an epitope of SEQ ID NO:4 or a glycopeptide derivative thereof.  
     
     
         52 . The kit of  claim 51 , wherein the peptide comprising an epitope of SEQ ID NO:4 is about 2 to 15 amino acids in length.  
     
     
         53 . The kit of  claim 45 , wherein the antibody binds immunologically to an epitope of SEQ ID NO:5 or a glycopeptide derivative thereof.  
     
     
         54 . The kit of  claim 53 , wherein the peptide comprising an epitope of SEQ ID NO:5 is about 2 to 15 amino acids in length.  
     
     
         55 . The kit of  claim 45 , wherein the antibody binds immunologically to an epitope of SEQ ID NO:5 or a glycopeptide derivative thereof.  
     
     
         56 . The kit of  claim 55 , wherein the peptide comprising an epitope of SEQ ID NO:5 is about 2 to 15 amino acids in length.  
     
     
         57 . The kit of  claim 45 , wherein the antibody binds immunologically to an epitope of SEQ ID NO:6 or a glycopeptide derivative thereof.  
     
     
         58 . The kit of  claim 55 , wherein the peptide comprising an epitope of SEQ ID NO:6 is about 2 to 15 amino acids in length.  
     
     
         59 . The kit of  claim 45 , wherein the antibody is a monoclonal antibody.  
     
     
         60 . The kit of  claim 45 , wherein the antibody is a polyclonal antibody.  
     
     
         61 . The kit of  claim 45 , wherein the antibody is labeled.  
     
     
         62 . A composition comprising antibodies or binding portions thereof, wherein said antibodies or binding portions bind to a peptide moiety, sugar moiety, or glycopeptide moiety of a urinary bladder antiproliferative factor (APF), wherein said APF comprises one or more sugar moieties.  
     
     
         63 . The composition of  claim 62 , wherein the antibodies or binding portions bind to a peptide moiety, a sugar moiety, or a glycopeptide moiety of the antiproliferative factor and inhibit activity of the APF molecule thereby.  
     
     
         64 . The composition of  claim 62 , wherein the peptide is SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, or SEQ ID NO:6.  
     
     
         65 . The composition of  claim 62 , wherein the antibodies are polyclonal antibodies.  
     
     
         66 . The composition of  claim 62 , wherein the antibodies are monoclonal antibodies.  
     
     
         67 . A method of detecting a bladder disorder in an individual, comprising the steps of: 
 obtaining a sample from the individual; and    assaying said sample for the presence of the composition of  claim 1 .    
     
     
         68 . The method of  claim 67 , wherein the bladder disorder is interstitial cystitis.  
     
     
         69 . The method of  claim 67 , wherein the assaying step is non-invasive or invasive to the individual.  
     
     
         70 . The method of  claim 67 , wherein the sample comprises urine, plasma, serum, tissue, or a mixture thereof from the individual.  
     
     
         71 . The method of  claim 67 , wherein the assaying step comprises detection of one or more sugar moieties of the composition of  claim 1 .  
     
     
         72 . The method of  claim 67 , wherein the assaying step comprises detection of the hydrophobic moiety of the composition of  claim 1 .  
     
     
         73 . The method of  claim 67 , wherein the antiproliferative factor is further defined as comprising a glycopeptide moiety.  
     
     
         74 . The method of  claim 73 , wherein the hydrophobic moiety of the composition comprises a peptide moiety and the assaying step is further defined as comprising introduction to the sample of an antibody directed to an epitope of a peptide, sugar, or glycopeptide moiety of a composition of  claim 1 .  
     
     
         75 . The method of  claim 67 , wherein the assaying sample comprises ELISA, Western blot, immunoblot, or radioimmunoassay.  
     
     
         76 . A method of treating an individual for cancer, comprising the step of administering to the individual a therapeutically effective amount of the composition of  claim 1 .  
     
     
         77 . The method of  claim 76 , wherein the cancer comprises an epithelial cancer.  
     
     
         78 . The method of  claim 76 , wherein the cancer comprises bladder cancer.  
     
     
         79 . The method of  claim 76 , wherein the cancer comprises prostate cancer.  
     
     
         80 . A method of treating a bladder disorder in an individual, comprising the step of administering to the individual a therapeutically effective amount of an inhibitor of the composition of  claim 1 .  
     
     
         81 . The method of  claim 80 , further comprising an additional bladder disorder therapy.  
     
     
         82 . The method of  claim 80 , wherein the bladder disorder comprises interstitial cystitis.  
     
     
         83 . The method of  claim 82 , further comprising an additional interstitial cystitis treatment and said administering of the inhibitor enhances the interstitial cystitis treatment.  
     
     
         84 . The method of  claim 83 , wherein the composition is administered prior to the interstitial cystitis treatment being enhanced, concomitant with the interstitial cystitis treatment being enhanced, subsequent to the interstitial cystitis treatment being enhanced, or a combination thereof.  
     
     
         85 . A method of treating a hyperplasia, comprising the step of administering a therapeutically effective amount of a composition of  claim 1 .  
     
     
         86 . A method of enhancing cancer treatment of an individual, comprising administering to the individual a therapeutically effective amount of a composition of  claim 1 .  
     
     
         87 . The method of  claim 86 , wherein administration of the composition enhances chemotherapy, radiotherapy, immunotherapy, surgery, gene therapy, or a combination thereof.  
     
     
         88 . The method of  claim 86 , wherein the composition is administered prior to the cancer treatment being enhanced, concomitant with the cancer treatment being enhanced, subsequent to the cancer treatment being enhanced, or a combination thereof.