High-Throughput Assay for Sugar-Mediated Drug Transport

Abstract

The invention provides a rapid, quantitative assay to directly assess the impact of a diverse range of sugars upon the sugar-mediated uptake of corresponding sugar-conjugates into various cell types.

1 . A method of assaying tumor selectivity for sugar-mediated transport of glycoconjugates contained in a glycorandomized library, comprising:
 (a) providing a glycorandomized library containing glycoconjugates differing by attached sugars;   (b) contacting each of said glycoconjugates with cells of a first line under conditions to allow uptake of the glycoconjugates into the cells; and   (c) assessing uptake of said glycoconjugates into the cells wherein sugar-mediated transport of the glycoconjugates into the cells is quantified relative to a corresponding conjugate lacking the attached sugars;   (d) repeating steps (a)-(c) with cells of a second, differing cell line; and   (e) comparing sugar-mediated transport of said glycoconjugates between cells of the first line and the second cell line,   wherein cell line selectivity for sugar-mediated transport of said glycoconjugates is indicated by elevated sugar transport activity in one of the cell lines relative to the other cell line.   
     
     
         2 . The method of  claim 1  wherein the cell line selectivity for sugar-mediated transport of said glycoconjugates of one cell line is at least three times higher relative to the other cell line. 
     
     
         3 . The method of  claim 1  wherein the glycoconjugates are contacted with cells independently and in parallel. 
     
     
         4 . The method of  claim 1  wherein the cells are cancerous. 
     
     
         5 . The method of  claim 1  wherein the glycoconjugates are sugar-fluorophore conjugates. 
     
     
         6 . The method of  claim 5  wherein the sugar-fluorophore is selected from the group consisting of aminofluoroscein, nitrobenzoxadiazole or dansyl chloride. 
     
     
         7 . A kit for assaying sugar-mediated transport of glycoconjugates contained in a glycorandomized library, comprising:
 (a) a glycorandomized library comprising glycoconjugates with different attached sugars;   (b) cells of a first line; and   (c) instructions for use.   
     
     
         8 . A kit for assaying tumor selectivity for sugar-mediated transport of glycoconjugates contained in a glycorandomized library, the kit comprising
 (a) a glycorandomized library containing glycoconjugates differing by attached sugars;   (b) cells of a first line;   (c) cells of a second, differing line; and   (d) instructions for use.