In situ binary synthesis of biologically effective molecules

Abstract

A plurality of oligonucleotides are caused to bind specifically to target nucleic acid which is predictive of a disease state or a biological condition within cells containing the target nucleic acid. The respective oligonucleotides, which may be functionilized or may be present in the form of oligonucleotide analogs, carry with them a plurality of synthons. Such synthons, which may be identiphores, toxiphores, or other precursors to biologically effective molecules, interact when specific binding of the respective oligonucleotides occurs at sites adjacent to each other on the target nucleic acid. The resulting interaction gives rise to the synthesis, generation or release of highly active biological molecules in situ in the cell in which the specific binding takes place. This permits the use of extraordinarily toxic molecules for use in killing cells containing the target nucleic acids. Imaging and other uses are also provided by the present invention.

What is claimed is:  
     
         1 . A method for identifying cells containing a preselected nucleic acid sequence comprising: 
 a. contacting said cells with a first oligonucleotide specifically bindable with said preselected nucleic acid, said first oligonucleotide comprising a first identiphore;    b. contacting said cells with a second oligonucleotide specifically bindable with said preselected nucleic acid, said second oligonucleotide comprising a second identiphore;    c. said first and second identiphores being detectable when in proximity with each other.    
     
     
         2 . The method of  claim 1  wherein said oligonucleotides orient said identiphores into spatial proximity when bound to said nucleic acid.  
     
     
         3 . The method of  claim 1  wherein said oligonucleotides orient said identiphores into a preselected geometric configuration when bound to said nucleic acid.  
     
     
         4 . The method of  claim 1  wherein said identiphores form a charge transfer complex when in proximity with each other.  
     
     
         5 . The method of  claim 1  wherein said identiphores become covalently bonded upon being placed into proximity.  
     
     
         6 . The method of  claim 1  wherein said detection comprises X-ray, magnetic resonance, proton magnetic resonance, electron spin resonance, fluorescence, or electromagnetic spectroscopy.  
     
     
         7 . The method of  claim 1  wherein the presence of said preselected nucleic acid is diagnostic for a cellular biological state.  
     
     
         8 . The method of  claim 7  wherein said cellular biological state is a disease state.  
     
     
         9 . The method of  claim 7  wherein said cellular biological state is a hyperproliferative state.  
     
     
         10 . The method of  claim 7  wherein said cellular biological state is a cancerous state.  
     
     
         11 . The method of  claim 1  wherein said identiphores form a new molecular species upon coming into proximity with each other.  
     
     
         12 . A method of killing cells containing a preselected nucleic acid sequence comprising: 
 contacting said cells with 
 a first oligonucleotide specifically bindable with said preselected nucleic acid, said first oligonucleotide comprising a first toxiphore;  
 and a second oligonucleotide specifically bindable with said preselected nucleic acid, said second oligonucleotide comprising a second toxiphore;  
   said first and second toxiphores forming a toxin upon coming into proximity with each other;    said first and second oligonucleotides being specifically bindable to adjacent sites on the preselected nucleic acid.    
     
     
         13 . The method of  claim 12  wherein said oligonucleotides orient said toxiphores into spatial proximity when bound to said nucleic acid.  
     
     
         14 . The method of  claim 12  wherein said oligonucleotides orient said toxiphores into a preselected geometric configuration when bound to said nucleic acid.  
     
     
         15 . The method of  claim 12  wherein said toxiphores become covalently bonded upon being placed into proximity.  
     
     
         16 . The method of  claim 12  wherein the presence of said preselected nucleic acid is predictive of a cellular biological state.  
     
     
         17 . The method of  claim 16  wherein said cellular biological state is a disease state.  
     
     
         18 . The method of  claim 16  wherein said cellular biological state is a hyperproliferative state.  
     
     
         19 . The method of  claim 16  wherein said cellular biological state is a cancerous state.  
     
     
         20 . The method of  claim 12  wherein said toxiphores form a new molecular species upon coming into proximity with each other.  
     
     
         21 . The method of  claim 20  wherein said toxin is the functional domain of a peptide.  
     
     
         22 . The method of  claim 21  wherein said peptide comprises the functional domain of ricin, sarcin, diphtheria toxin, or botulism toxin.  
     
     
         23 . The method of  claim 20  wherein said toxin comprises the functional domain of a bacterial toxin.  
     
     
         24 . The method of  claim 20  wherein said toxin is an invertebrate toxin.  
     
     
         25 . The method of  claim 24  wherein said toxin is an arachnid toxin.  
     
     
         26 . The method of  claim 20  wherein said toxin is an amphibian toxin.  
     
     
         27 . The method of  claim 12  wherein fewer than ten molecules of said toxin is sufficient to kill each of said cells.  
     
     
         28 . The method of  claim 12  wherein said contacting is under nucleic acid heteroduplex forming conditions.  
     
     
         29 . The method of  claim 12  wherein said contacting is of a mammalian tissue.  
     
     
         30 . A method for the in situ synthesis of a chemical species comprising 
 specifically binding first and second oligonucleotides to a preselected nucleic acid at adjacent sites thereof;    said first and second oligonucleotides comprising first and second synthons.    
     
     
         31 . The method of  claim 30  wherein said first and second synthons form said chemical species upon coming into proximity with each other.  
     
     
         32 . The method of  claim 31  wherein said proximity comprises spatial proximity and geometric proximity.  
     
     
         33 . The method of  claim 30  wherein said chemical species is toxic to cells containing the nucleic acid.  
     
     
         34 . The method of  claim 30  wherein said chemical species is detectible in cells containing the nucleic acid.  
     
     
         35 . The method of  claim 30  wherein said preselected nucleic acid is predictive of a biological state in cells containing said nucleic acid.  
     
     
         36 . The method of  claim 35  wherein said biological state is a disease state.  
     
     
         37 . The method of  claim 35  wherein said biological state is hyperproliferation.  
     
     
         38 . The method of  claim 35  wherein said biological state is a neoplastic state.  
     
     
         39 . The method of  claim 30  wherein the first and second synthons are brought into spatial and geometric proximity upon the specific binding of the oligonucleotides to the preselected nucleic acid.  
     
     
         40 . A pair of oligonucleotides: 
 each of said oligonucleotides being specifically bindable with a preselected nucleic acid at adjacent binding sites;    each of said oligonucleotides comprising a portion of a molecular species, which species is formed when said adjacent, specific binding of the oligonucleotides to the nucleic acid occurs.    
     
     
         41 . The pair of  claim 40  wherein said molecular species is a toxin.  
     
     
         42 . The pair of  claim 40  wherein said molecular species is a charge transfer complex.  
     
     
         43 . The pair of  claim 40  wherein said molecular species is a detectable molecule.  
     
     
         44 . The pair of  claim 40  wherein said molecular species is a cell regulatory moiety.  
     
     
         45 . A pharmaceutical comprising the pair of oligonucleotides of  claim 40  in a pharmaceutically acceptable carrier or diluent.