Antisense inhibition of angiogenin expression

Abstract

Disclosed are oligonucleotide compounds that inhibit the expression of angiogenin when administered to a mammal. Also disclosed are methods and pharmaceutical compositions for inhibiting the expression of angiogenin useful in therapy or diagnosis.

What is claimed is:  
     
         1 . A compound for inhibiting expression of angiogenin comprising an oligonucleotide or analog thereof having a base sequence complementary to a target portion of a nucleic acid encoding angiogenin.  
     
     
         2 . The compound of  claim 1  wherein the base sequence is configured to bind to the target portion of the nucleic acid in a manner to inhibit the expression of angiogenin.  
     
     
         3 . The compound of  claim 2  wherein the oligonucleotide analog comprises a modified internucleotide linkage, a modified purine or pyrimidine moiety, a modified sugar moiety, a modified 5′ hydroxyl moiety, a modified 3′ hydroxyl moiety or a modified 2′ hydroxyl moiety.  
     
     
         4 . The compound of  claim 3  wherein the modified internucleotide linkage comprises a substituent having an improved aqueous or lipid solubility or improved resistance to nuclease digestion.  
     
     
         5 . The compound of  claim 4  wherein the modified internucleotide linkage is selected from the group consisting of phosphorothioate, alkyl or cycloalkyl phosphorothioate, N-alkyl or cycloalkyl phosphoramidates, phosphorodithioates, alkyl or cycloalkyl phosphonates, phosphodiester, phosphotriester, C 1 - C 4  alkyl, cycoaloyl, short chain heteroatomic or heterocyclic backbone, morpholino backbone, polyprotein-nucleic acid or peptide-nucleic acid backbone, polyamide, CH 2 —NH—O—CH 2 , CH 2 —N(CH 3 )—O—CH 2 , CH 3 —O—N(CH 3 )—CH 2 , CH 2 —N(CH 3 )—CH 2  and O—N(CH 3 )—CH 2 —CH 2 .  
     
     
         6 . The compound of  claim 3  wherein the modified purine or pyrimidine moiety includes inosine.  
     
     
         7 . The compound of  claim 3  wherein the modified sugar moiety includes sugar mimetics comprising C 4 -C 8  cycloalkyl.  
     
     
         8 . The compound of  claim 3  wherein the modified 5′ or 3′ hydroxyl moiety is selected from the group consisting of C 1-4  alkoxy, intercalating agent, peptide, enzyme, ribozyme, substituted acridine, 2-methoxy-6-chloro-9-pentylaminoacridine, N-(6-chloro-2-methoxyacridinyl)-0-methoxydisopropylaminophosphinyl-3-aminopropanol and N-(6 chloro-2-methoxyacridinyl)-O-methoxydisopropylaminophosphinyl-5-aminopentanol.  
     
     
         9 . The compound of  claim 1  wherein the modified 2′ hydroxyl moiety is selected from the group consisting of OH, SH, SCH 2 , OCH 3 , F, OCN, OCH 6 CH 3 , OCH 3 OCH 3 , OCH 3 O(CH 2 ) n CH 3 , O(CH 2 ) n NH 2  or O (CH 2 ) n CH 3  where n is from 1 to about 10; C 1  to C 10  lower allyl, substituted lower alkyl, alkaryl or aralkyl; Cl; Br; CN; CF 3 ; OCF 3 ; O, S, or N-alkyl; O, S, or N-alkenyl; SOCH 3 ; SO 2 CH 3 ; ONO 2 ; NO 2 ; N 3 ; NH 2 ; heterocycloalkyl or alkaryl; aminoalkylamino; polyalkylamino; substituted silyl: an RNA cleaving group; a cholesteryl group; a conjugate; a reporter group; an intercalator; a group for improving the pharmacokinetic properties of an oligonucleotide; and a group for improving the pharmacodynamic properties of an oligonucleotide.  
     
     
         10 . The compound of  claim 1  wherein the base sequence of the oligonucleotide or analog thereof is selected from the group consisting of  
       
         
           
                 
               
                     
                 
                   5′-GCCCATCACCATCTCTTC-3′, 
                 
                     
                 
                   5′-ACACGGCATCATGAATCA-3′, 
                 
                     
                 
                   5′-CCAGCGGCCCGCTGGTTA-3′, 
                 
                     
                 
                   5′-ACCAAATTTTATATTCTA-3′, 
                 
                     
                 
                   5′-CAGGCCCATCACCATCAC-3′, 
                 
                     
                 
                   5′-GCCCAGGCCCATCACCAT-3′, and 
                 
                     
                 
                   5′-TCTCTGACACGGCATCAT-3′. 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         11 . A composition for inhibiting expression of angiogenin comprising an effective amount of an oligonucleotide or analog thereof having a base sequence complementary to a target portion of a nucleic acid encoding angiogenin in a pharmaceutically acceptable carrier.  
     
     
         12 . The composition of  claim 11  wherein the base sequence of the oligonucleotide or analog thereof is selected from the group consisting of  
         
     
     
         13 . A compound for inhibiting expression of angiogenin having the  
       
         
           
           
               
               
           
         
       
       wherein 
 X is O, S, or C 1-4  alkyl;  
 B is adenine, guanine, cytosine, or thymine selected such that the oligonucleotide has a complementary base sequence with a portion of a target nucleic acid strand coding for angiogenin thereby inhibiting expression thereof;  
 R 1  is H, C 1-4  alkyl, intercalating agent, peptide, enzyme, ribozyme, substituted acridine, 2-methoxy-6-chloro-9-pentylaminoacridine, N-(6-chloro-2-methoxyacridinyl)-O-methoxydisopropylaminophosphinyl-3-aminopropanol and N-(6 chloro-2-methoxyacridinyl)-O-methoxydisopropylaminophosphinyl-5-aminopentanol. or substituted acridine;  
 R 2  is H, OH, SH, SCH 2 , OCH 3 , F, OCN, OCH 6 CH 3 , OCH 3 OCH 3 , OCH 3 O(CH 2 ) n CH 3 , O(CH 2 ) n NH 2  or O(CH 2 ) n CH 3  where n is from 1 to about 10; C 1  to C 10  lower alkyl, substituted lower alkyl, alkaryl or aralkyl; Cl; Br; CN; CF 3 ; OCF 3 ; O, S, or N-alkyl; O, S, or N-alkenyl; SOCH 3 ; SO 2 CH 3 ; ONO 2 ; NO 2 ; N 3 ; NH2; heterocycloalkyl or alkaryl; aminoalkylamino; polyalkylamino; substituted silyl: an RNA cleaving group; a cholesteryl group; a conjugate; a reporter group; an intercalator; a group for improving the pharmacokinetic properties of an oligonucleotide; or a group for improving the pharmacodynamic properties of an oligonucleotide; and  
 n is 5to 100.  
 
     
     
         14 . The compound of  claim 13  wherein the base sequence is selected from the group consisting of  
       
         
           
                 
               
                     
                 
                   5′-ACACGOCATCATGAATCA-3′, 
                 
                     
                 
                   5′-CCAGGGOCCCGCTGGTTA-3′, 
                 
                     
                 
                   5′-ACCAAATTTTATATTCTA-3′, 
                 
                     
                 
                   5′-CAGOCCCATCACCATCAC-3′, 
                 
                     
                 
                   5′-GCCCAG6CCCATCACCAT-3′, and 
                 
                     
                 
                   5′-TCTCTGACACGGCATCAT-3′. 
                 
                     
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         15 . A method for inhibiting expression of angiogenin in a mammal comprising administering to the mammal an effective amount of an oligonucleotide or analog thereof having a base sequence complementary to a target portion of a nucleic acid encoding angiogenin so as to inhibit the expression of angiogenin.  
     
     
         16 . A method for reducing size of tumors associated with angiogenesis in a mammal comprising administering to the mammal an effective amount of an oligonucleotide or analog thereof having a base sequence complementary to a target portion of a nucleic acid encoding angiogenin so as to reduce tumor size.  
     
     
         17 . A method for decreasing production of angiogenin in a mammal comprising administering to the mammal an effective amount of an oligonucleotide or analog thereof having a base sequence complementary to a target portion of a nucleic acid encoding angiogenin so as to decrease production of angiogenin.  
     
     
         18 . A method for inhibiting metastasis of tumor cells in a mammal comprising administering to the mammal an effective amount of an oligonucleotide or analog thereof having a base sequence complementary to a target portion of a nucleic acid encoding angiogenin so as to inhibit metastasis of tumor cells.  
     
     
         19 . A method for inhibiting the establishment of tumor cells in a mammal comprising administering to the mammal an effective amount of an oligonucleotide or analog thereof having a base sequence complementary to a target portion of a nucleic acid encoding angiogenin so as to inhibit establishment of tumor cells.  
     
     
         20 . A method for inhibiting growth of tumors associated with angiogenesis in a mammal comprising administering to the mammal an effective amount of an oligonucleotide or analog thereof having a base sequence complementary to a target portion of a nucleic acid encoding angiogenin so as to inhibit tumor growth.  
     
     
         21 . A method for detecting the presence of angiogenin in a sample comprising contacting the sample with a labeled oligonucleotide or analog thereof having a base sequence complementary to a target portion of a nucleic acid encoding angiogenin; 
 allowing the labeled oligonucleotide or analog thereof to bind to the target portion of the nucleic acid encoding angiogenin; and    detecting the labeled oligonucleotide or analog thereof.    
     
     
         22 . A method for detecting the presence of angiogenin in a mammal comprising administering to the mammal a labeled oligonucleotide or analog thereof having a base sequence complementary to a target portion of a nucleic acid encoding angiogenin; 
 allowing the labeled oligonucleotide or analog thereof to bind to the target portion of the nucleic acid encoding angiogenin; and    detecting the labeled oligonucleotide or analog thereof.    
     
     
         23 . A method for diagnosing conditions associated with abnormal angiogenesis in a mammal comprising administering to the mammal a labeled oligonucleotide or analog thereof having a base sequence complementary to a target portion of a nucleic acid encoding angiogenin; 
 allowing the labeled oligonucleotide or analog thereof to bind to the target portion of the nucleic acid encoding angiogenin;    detecting the labeled oligonucleotide or analog thereof;    measuring the labeled oligonucleotide or analog thereof; and    determining the abnormal condition based on the detecting and measuring of the labeled oligonucleotide or analog thereof.