Multifunctional Poly(amine-co-ester-co-orthoester) for Efficient and Safe Gene Delivery

Abstract

Cationic polymers are used for non-viral gene delivery, but current materials lack the functionality to address the multiple barriers involved in gene delivery. Here we describe the rational design and synthesis of a new family of quaterpolymers with unprecedented multifunctionality: acid sensitivity, low cationic charge, high hydrophobicity, and biodegradability, all of which are essential for efficient and safe gene delivery. The polymers were synthesized via lipase-catalyzed polymerization of ortho ester diester, lactone, dialkyl diester, and amino diol monomers. Polymers containing ortho ester groups exhibited acid-sensitive degradation at endosomal pH (4~5), facilitated efficient endosomal escape and unpackaging of the genes, and were efficient in delivering genetic materials to HEK293 cells, human glioma cells, primary mouse melanoma cells, and human umbilical vein endothelial cells (HUVECs). We also developed a highly efficient lyophilized formulation of the nanoparticles, which could be stored for a month without loss of efficiency.

Excipients for the lyoprotection of MAPKAP kinase 2 inhibitory peptide nano-polyplexes

“…There is also precedent for polyplexes lyophilized with excipients having improved activity over the freshly prepared parent formulation. For example, Zhang et al demonstrated a trehalose dose-dependent increase in transfection efficiency for lyophilized poly(amine-co-ester-co-orthoester) (oPACE) polyplexes for gene delivery, with polyplexes lyophilized in 158 mM trehalose nearly doubling the transfection efficiency relative to fresh oPACE-pDNA polyplexes [37]. Thus, our results suggest that results from polyplexes used for gene delivery are relevant and can be extrapolated to the PPAA-based polyplexes designed for delivery of cationic peptides.…”

Recent advances in the synthesis of biodegradable polyesters by sustainable polymerization: lipase-catalyzed polymerization

“…The same group synthesized a series of poly(amine-co-ester-co-orthoester) with acid sensitivity and redox-responsive, these polyesters have been proved to be gene delivery and nanocarriers for doxorubicin delivery. 93,98,101 Similarly, Liu et al developed the terpolymerization cationic poly(amine-co-ester) for DNA condensation with PEG shell for nanoparticle stabilization and poly(glutamic acid) and mTAT (a cell-penetrating peptide) for accelerating cellular uptake, and a nuclear localization signal peptide for enhanced intracellular transport of DNA to the nucleus. 102 Such an approach presents a more sustainable and environmentally friendly synthesis and could also be used for controlled drug release and gene delivery.…”

Debugging the genetic code: Non-viral in vivo delivery of therapeutic genome editing technologies

“…New lipids and polymers have been developed with the goal of improving nucleic acid delivery in vitro and in vivo , and these could prove useful for delivering editing agents. For example, poly(amine-co-ester) terpolymers have been used to successfully deliver pDNA in vivo [47, 48], and have recently been shown to effectively deliver TFO PNAs ex vivo (A.S. Piotrowski-Daspit et al, abstract 614, 21 st Annual Meeting of American Society of Gene & Cell Therapy, Chicago IL, May 2018). Additionally, the incorporation of targeting moieties such as antibodies for specific cell types could be useful particularly for local delivery.…”