Modified antisense nucleotides complementary to a section of the human haras gene

Abstract

The invention relates to a specific modified oligonucleotide complementary to a section of the human Ha-ras gene and mRNA, and its use to specifically regulate, modulate or inhibit expression of the HA-ras gene, and its use as a pharmaceutical for the treatment of conditions arising from the abnormal expression of the Ha-Ras gene, in particular in combination with chemotherapy and radiotherapy. The modified oligodeoxynucleotide according to the invention has the sequence 5′-TxAxTxTxCxCxGxTxCxAxT-3′-O—PO 2 —O—R (SEQ ID NO:1), wherein X is an internucleotide linkage of type o or s, with the proviso that x is an s linkage at least 4 times and at most 9 times, and o means a phosphodiester internucleoside linkage, s means a phosphorothioate internucleoside linkage, R means a C 8 -C 21 alkyl group, —(CH 2 —CH 2 O)n-(CH 2 ) m —CH 3 , or —CH 2 —CH(OH)CH 2 O—(CH 2 ) q —CH 3 wherein n is an integer from 1 to 6, m is an integer from 0 to 20 and q is an integer from 7 to 20 and A is 2′-deoxyadenosine, G is 2′-deoxyguanosine, C is 2′-deoxycytidine and T is thymidine.

1 - 45 . (Cancelled)  
     
     
         46 . An oligodeoxynucleotide comprising SEQ ID NO:1 (5′-TxAxTxTxCxCxGxTxCxAxT-3′-O—PO 2 —O—R), wherein 
 x is an internucleotide linkage of type o or s, with the proviso that x is an s linkage at least 4 times and at most 9 times; 
 o means a phosphodiester internucleoside linkage; and  
 s means a phosphorothioate internucleoside linkage;  
   R means a C 8 -C 2 , alkyl group, —(CH 2 —CH 2 O) n —(CH 2 ) m —CH 3  or —CH 2 —CH(OH)CH 2 O—(CH 2 ) q —CH 3 , wherein n is an integer from 1 to 6, m is an integer from 0 to 20, and q is an integer from 7 to 20;    A is 2′-deoxyadenosine;    G is 2′-deoxyguanosine;    C is 2′-deoxycytidine; and    T is thymidine.    
     
     
         47 . A method of treating a disease arising from the overexpression and/or mutation of the Ha-ras gene, said method comprising administering to a host an effective amount of the oligonucleotide of  claim 46 .  
     
     
         48 . A method of treating a disease arising from a hyperproliferative disorder, said method comprising administering to a host an effective amount of the oligodeoxynucleotide of  claim 46 .  
     
     
         49 . The method of  claim 48 , wherein the disease is cancer.  
     
     
         50 . The method of  claim 49 , further comprising administering radiotherapy.  
     
     
         51 . The method of  claim 49 , further comprising administering at least one chemotherapeutic agent.  
     
     
         52 . The method of  claim 48 , wherein the disease is restenosis.  
     
     
         53 . The method of  claim 48 , wherein the disease is psoriasis.  
     
     
         54 . A method of regulation, modulation, or inhibition of Ha-ras gene expression, said method comprising administering to a host an effective amount of the oligodeoxynucleotide of  claim 46 .  
     
     
         55 . A method of inhibiting the proliferation of cancer cells, said method comprising administering to a host an effective amount of the oligodeoxynucleotide of  claim 46 .  
     
     
         56 . A method of reversing the radio-resistance of cancer cells, said method comprising administering to a host an effective amount of the oligodeoxynucleotide of  claim 46 .  
     
     
         57 . A method of reversing the chemo-resistance of cancer cells, said method comprising administering to a host an effective amount of the oligodeoxynucleotide of  claim 46 .  
     
     
         58 . A method of preparing the oligodeoxynucleotide of  claim 46 , said method comprising forming phosphorothioate linkages between nucleotides of the oligodeoxynucleotide by sulfurization using Beaucage reagent.