Ca ix - nir dyes and their uses

Abstract

The present disclosure relates to compounds that are useful as near-infrared fluorescence probes, wherein the compounds include i) a ligand that binds to the active site of carbonic anhydrase, ii) a dye molecule, and iii) a linker molecule that comprises an amino acid, amide, ureido, or polyethylene glycol derivative thereof. The disclosure further describes methods and compositions for making and using the compounds, methods incorporating the compounds, and kits incorporating the compounds.

The invention claimed is: 
     
         1 . A composition comprising a first compound, or salt thereof and a second compound, or salt thereof, wherein the first compound has a formula
                       wherein B is a CA IX-targeted molecule is selected from the group consisting of
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
                     
 and 
                     
   wherein n is 0, 1, 2, or 3; m is 0, 1, 2, 3, 4, or 5, and p is 0, 1, 2, or 3;
 W comprises an extended hydrophobic residue; 
 X comprises a hydrophobic spacer; 
 Y comprises an amino acid spacer; and 
 Z comprises a NIR dye; 
 and wherein the second compound comprises an NIR dye and pteroyl ligand conjugated to an amino acid linking group or a PSMA-targeting compound conjugated to an amino acid linking group. 
   
     
     
         2 . The composition of  claim 1 , further comprising a third compound, wherein the second compound comprises an NIR dye and a pteroyl ligand conjugated to an amino acid linking group and the third compound comprises an NIR dye and PSMA-targeting compound conjugated to an amino acid linking group. 
     
     
         3 . The composition of  claim 1 , wherein W is a hydrophobic amino acid, aromatic amino acid, alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, methionine, tyrosine, arginine, histidine, lysine, glycine, serine, threonine, cysteine, tyrosine, asparagine, aspartic acid, glutamic acid, glutamine and derivatives thereof. 
     
     
         4 . The composition of  claim 1 , wherein X is a length from 7 atoms to 14 atoms. 
     
     
         5 . The composition of  claim 1 , wherein X is selected from the group consisting of polyethylene glycol (PEG), polyethylene amine (PEA), N-amino-dPEG 2  acid, a chain of 6 atoms, a spacer 6 atoms in length, a chain from 6 to 20 atoms in length, and a peptide comprising aryl or aryl alkyl groups, each of which is optionally substituted, and wherein one aryl or aryl alkyl group is about 6 to about 10, or about 6 to about 14 atoms, and the other aryl or aryl alkyl group is about 10 to about 14, or about 10 to about 15 atoms, or about 1 to about 20 atoms. 
     
     
         6 . The composition of  claim 1 , wherein X, Y, and Z, are independently selected from the group consisting of a variable charge, negative charge, and positive charge. 
     
     
         7 . The composition of  claim 1 , wherein X is a peptide compromising positively charged amino acids or quaternary amine containing amino acid. 
     
     
         8 . The composition of  claim 1 , wherein Y is selected from the group consisting of an aromatic amino acid, an amino acid spacer with a chalcogen-containing side chain group, an amino acid spacer with a sulfur-containing side chain group, an amino acid spacer with a sulfur-containing thiophenol moiety, a tyrosine with a carbon isotope on the aromatic ring of tyrosine, a an amino acid with an aromatic ring with a hydrogen isotope aspartic acid, glutamic acid, arginine, histidine, lysine, glycine, serine, threonine, cysteine, tyrosine, asparagine, aspartic acid, glutamic acid, glutamine, alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, methionine, and derivatives thereof. 
     
     
         9 . The composition of  claim 1 , wherein the NIR dye is selected from the group consisting of, IR800, SP054, S0121,-S2076, S0456,
                                                                                                                                                                                 and                         wherein R41, R42, R43, R44, R45, R46, R47, R48, = H or SO   3 H; X = O, S, or N. 
     
     
         10 . A method of optical imaging of a biological tissue of a subject, comprising administering the composition of  claim 1  or  claim 2  to a subject, wherein the compounds distribute within a biological target; illuminating biological tissue within the biological target with an excitation light of a wavelength absorbable by the compounds; and detecting the optical signals emitted by the compounds. 
     
     
         11 . The method of  claim 10 , wherein each compound has signal properties that are distinguishable from at least one other compound. 
     
     
         12 . The method of  claim 10 , wherein the optical signals emitted by the compounds are used to construct at least one image. 
     
     
         13 . The method of  claim 10 , wherein the illuminating and/or detecting steps are performed using an endoscope, catheter, tomographic system, hand-held optical imaging system, surgical goggles, or intra-operative microscope. 
     
     
         14 . A method of identifying a target cell type in a biological sample of a subject comprising administering the composition of  claim 1  or  claim 2  to a subject, wherein the compounds bind to at least one target cell type, and optically detecting the presence or absence of the compounds in the biological sample, wherein presence of the compound indicates that the target cell type is present in the biological sample. 
     
     
         15 . The method of  claim 14  wherein the target cell type is tissue to be imaged for diagnostic purposes. 
     
     
         16 . The method of  claim 15  wherein the tissue is a tumor or a lymph node. 
     
     
         17 . The method of  claim 15  wherein the target cell type is cancerous. 
     
     
         18 . The method of  claim 17  wherein the cancer is selected from the group consisting of brain, breast, cervical, rectal and lung cancer. 
     
     
         19 . A method of performing image guided surgery on a subject comprising administering the composition of  claim 1  or  claim 2  to the subject illuminating the compounds to visualize the compound using infrared light; and performing surgical resection of areas that fluoresce upon excitation by the infrared light. 
     
     
         20 . A method of diagnosing a disease in a subject comprising administering the composition of  claim 1  or  claim 2  to the subject wherein the compounds bind to at least one target cell type, measuring an optical signal from the compounds present in the target cell type; comparing the optical signal measured with at least one control data set, wherein the at least one control data set comprises optical signals from the compounds contacted with a biological sample that does not comprise the target cell type; and providing a diagnosis of disease wherein the comparison indicates the presence of the disease.