A Novel PEGylation Method for Improving the Pharmacokinetic Properties of Anti-Interleukin-17A RNA Aptamers

Abstract

The obstacles to the development of therapeutic aptamers for systemic inflammatory diseases, such as nuclease degradation and renal clearance, have not been fully overcome. Here, we report a novel PEGylation method, sbC-PEGylation, which improves the pharmacokinetic properties of RNA aptamers that act against interleukin-17A (IL-17A) in mice and monkeys. sbC-PEGylated aptamers were synthesized by coupling the symmetrical branching molecule 2-cyanoethyl-N,N-diisopropyl phosphoroamidite to the 5′ end of the aptamer, before conjugating two polyethylene glycol (PEG) molecules to the aptamer. Pharmacokinetic studies showed that compared with conventionally PEGylated aptamers, the sbC-PEGylated aptamer exhibited excellent stability in the blood circulation of mice and monkeys. In addition, one of the sbC-PEGylated aptamers, 17M-382, inhibited the interleukin-6 (IL-6) production induced by IL-17A in NIH3T3 cells in a concentration-dependent manner, and the half-maximal inhibitory concentration of sbC-PEGylated 17M-382 was two times lower than that of non-PEGylated 17M-382. Furthermore, the intraperitoneal administration of sbC-PEGylated 17M-382 significantly inhibited the IL-6 production induced by IL-17A in a mouse air pouch model. Our findings suggest that the novel PEGylation method described in this study, sbC-PEGylation, could be used to develop anti-IL-17A aptamers as a therapeutic option for systemic inflammatory disease.

A Synthetic Cross-Species CD200R1 Agonist Suppresses Inflammatory Immune Responses In Vivo

“…Modification with PEG has previously been proven to effectively improve the pharmacokinetic properties of DNA aptamers in vivo . 24 , 49 , 50 As projected, the PEGylated form of CCS13 (PEG-CCS13) retained its immunosuppressive function in both human and murine allo-MLC assays in a CD200R1-specific manner ( Figures 3 A and 3B, respectively). We subsequently assessed its capacity to suppress inflammatory responses in two distinct murine models of inflammation.…”

Using Aptamers as a Novel Method for Determining GnRH/LH Pulsatility

“…Post-SELEX, modifications of the aptamer structure can be employed to improve stability (e.g., by capping the 3′-end of the oligonucleotide with inverted thymidine or biotin to reduce its susceptibility to 3′-exonuclease [ 21 , 22 ]), increase the half-life of the aptamer in vivo (e.g., by conjugation of polyethylene glycol (PEG) to the 5′-end [ 23 ] to reduce the rate of renal clearance) and enhance ligand-binding affinity (e.g., by adding amino groups to the oligonucleotide sequence [ 24 ] and/or modifying nucleotide bases within the sequence [ 25 ]). More recently, other types of modified aptamers have been developed, with increased resistance to degradation by nucleases and/or increased binding affinities, including spiegelmers (oligonucleotide sequences consisting of L-(deoxy)ribonucleic acids that are synthetic mirror-image isomers of naturally-occurring D-(deoxy)ribonucleic acids) [ 26 ]; circular aptamers [ 27 ]; and multimers (i.e., aptamers that have the ability to bind to more than one region of their ligands, which improve the aptamers’ sensitivity and specificity) [ 28 , 29 ].…”

“…Modifications made to aptamers to improve their pharmacokinetic properties can also adversely affect their function. For instance, conjugation of PEG to an interleukin-17A RNA aptamer more than doubled the half-life of the aptamer in vivo [ 23 ]. However, significant titres of anti-PEG antibodies were detected in 37% of plasma samples from 377 healthy donors [ 88 ].…”

Aptamers as Diagnostic Tools in Cancer

“…To solve the problem of renal filtration, aptamers are generally linked to polyethylene glycol (PEG), cholesterol, or proteins. Using a multivalent aptamer design can also help to achieve the same goal [ 55 , 56 , 57 ]. Regarding toxicity, while it is very limited in humans, chemical modifications should be used carefully in order to avoid undesired effects.…”