Methods for increasing in vivo efficacy of oligonucleotides and inhibiting inflammation in mammals

Abstract

The invention relates to the use of nucleotide substitutes for increasing the in vivo efficacy of nucleic acid molecules and also for inhibiting inflammation in mammals. More particularly, the present invention relates to the use of 2′6′ diaminopurine (DAP) and analogs thereof per se in anti-inflammatory compositions, and also for preparing nucleic acid molecules having an increased in vivo physiological efficiency and a reduced toxicity as compared to conventional oligos. The invention is particularly useful for the preparation of antisense oligonucleotides for treating pulmonary/respiratory diseases such as cystic fibrosis, asthma, chronic bronchitis, chronic obstructive lung disease, eosinophilic bronchitis, allergies, allergic rhinitis, pulmonary fibrosis, adult respiratory distress syndrome, sinusitis, respiratory syncytial virus or other viral respiratory tract infection and cancer.

1 . An oligonucleotide comprising an antisense oligonucleotide against the common beta subunit of IL-3, IL-5 and GM-CSF, wherein the oligonucleotide comprises at least one adenosine and said adenosine has been replaced with a nucleotide substitute, wherein the nucleotide substitute is 2-amino-2′-deoxyadenosine or a salt thereof. 
     
     
         2 . The oligonucleotide of  claim 1 , wherein the nucleotide substitute is 2-amino-2′-deoxyadenosine. 
     
     
         3 . The oligonucleotide of  claim 2 , wherein said oligonucleotide is a DNA oligonucleotide. 
     
     
         4 . The oligonucleotide of  claim 2 , wherein said oligonucleotide is an RNA oligonucleotide. 
     
     
         5 . The oligonucleotide of  claim 2 , wherein the oligonucleotide is selected from the group consisting of SEQ ID NOs: 1-18, 23 and 26. 
     
     
         6 . The oligonucleotide of  claim 1 , wherein the oligonucleotide comprises SEQ ID NO. 8. 
     
     
         7 . The oligonucleotide of  claim 6 , wherein the oligonucleotide consists of SEQ ID NO. 8. 
     
     
         8 . The oligonucleotide of  claim 1 , wherein the oligonucleotide has at least one mononucleotide linking residue selected from the group consisting of methylphosphonate, phosphotriester, phosphorothioate, phosphodiester, phosphorodithioate, boranophosphate, formacetal, thioformacetal, thioether, carbonate, carbamate, sulfate, sulfonate, sulfamate, sulfonamide, sulfone, sulfite, sulfoxide, sulfide, hydroxylamine, methylene (methyimino), methyleneoxy (methylimino), and phosphoramidate residues. 
     
     
         9 . A pharmaceutical composition comprising the oligonucleotide of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         10 . A method for treating and/or preventing a disease, comprising administering a pharmaceutical composition of  claim 9  to a subject in need thereof, wherein the disease is selected from the group consisting of respiratory system diseases, neurological diseases, cardiovascular diseases, rheumatological diseases, digestive diseases, cutaneous diseases, ophtalmological diseases, urinary system diseases, cancers, pathogen infections, genetic diseases, hypereosinophilia, general inflammation, and cancers. 
     
     
         11 . The composition of  claim 9 , wherein said oligonucleotide is present in an amount of about 1% to about 90% of the composition. 
     
     
         12 . The composition of  claim 9 , further comprising an agent selected from the group consisting of drugs, antioxidants, surfactants, flavoring agents, volatile oils, buffering agents, dispersants, propellants, preservatives, and combinations thereof. 
     
     
         13 . The method of  claim 10 , wherein the disease is a respiratory system disease associated with an inflammation of the lungs, the airways and/or the nose. 
     
     
         14 . The method of  claim 13 , wherein the respiratory system disease is selected from the group consisting of pulmonary fibrosis, adult respiratory distress syndrome, cystic fibrosis, chronic obstructive lung disease, chronic bronchitis, eosinophilic bronchitis, asthma, allergy, allergic rhinitis, sinusitis and hypereosinophilia. 
     
     
         15 . The composition of  claim 9 , wherein the composition is contained in a pressurized aerosol dispenser, a nasal sprayer, a nebulizer, a metered dose inhaler, a dry powder inhaler, or a capsule.