Stem-loop compositions and methods for inhibiting interleukin-8
The application discloses methods and compositions for inhibiting functions associated with Interleukin-8 (IL8). The methods and compositions may involve the use of aptamers for binding to IL8 and preventing or reducing association of IL8 with CXCR1, CXCR2, or both. The methods and compositions may include one or more aptamers that bind to an N-terminal domain of IL8. The methods and compositions may include one or more aptamers that bind to a hydrophobic pocket of IL8. The methods and compositions may include one or more aptamers that bind to an N-loop of IL8. The methods and compositions may include one or more aptamers that bind to a GAG binding site of IL8. The application further provides anti-IL8 aptamers for the treatment of ocular diseases or disorders. In some cases, the anti-IL8 aptamers may have a stem-loop secondary structure.
1 . An aptamer that binds to and inhibits Interleukin-8 (IL8), comprising a secondary structure comprising at least one terminal loop comprising greater than three nucleotides, wherein said terminal loop selectively binds to an epitope of IL8, wherein said epitope is not a GAG-binding site and wherein the aptamer comprises a consensus nucleic acid sequence of 5′-NNYVANDDNWGWDDNNRGKNNGHGUGNHHNVRNN-3′ (SEQ ID NO:92), where N is A, C, G, or U; Y is Cor U; Vis A, C, or G; D is A, G, or U; W is A or U; R is A or G; K is G or U; and H is A, C, or U.
2 . The aptamer of claim 1 , wherein said secondary structure further comprises in a 5′ to 3′ direction: (i) a first base paired stem; (ii) a first loop; (iii) a second base paired stem; and (iv) a second loop.
3 . The aptamer of claim 1 , wherein said secondary structure comprises in a 5′ to 3′ direction: (i) a first base paired stem; (ii) a first loop; (iii) a second base paired stem; (iv) a second loop; (v) a third base paired stem; (vi) a third loop; and (vii) a fourth loop.
4 . The aptamer of claim 3 , wherein said first loop comprises a nucleic acid sequence of 5′-A-3′.
5 . The aptamer of claim 3 , wherein said second loop comprises a nucleic acid sequence of 5′-AG-3′.
6 . The aptamer of claim 3 , wherein said fourth loop comprises a nucleic acid sequence of 5′-G-3′.
7 . The aptamer of claim 1 , wherein said terminal loop comprises a nucleic acid sequence that selectively binds to a N-terminal domain of Interleukin-8 (IL8), a hydrophobic pocket of IL8, a N-loop of IL8, or any combination thereof.
8 . The aptamer of claim 1 , wherein said aptamer comprises RNA, modified RNA or a combination thereof.
9 . The aptamer of claim 1 , wherein said aptamer comprises one or more modified nucleotides.
10 . The aptamer of claim 1 , wherein said aptamer comprises a nuclease-stabilized nucleic acid backbone.
11 . The aptamer of claim 1 , wherein said aptamer is conjugated to a polyethylene glycol (PEG) molecule.