Multifunctional Neomycin-Triazine-Based Cationic Lipids for Gene Delivery with Antibacterial Properties.

Chiara Pennetta,Fiorenza Viani,Alessandro Volonterio,Maria Cristina Bellucci,Federica Ponti,Nina Bono,Gabriele Candiani

doi:10.1021/acs.bioconjchem.0c00616

Chiara Pennetta, Fiorenza Viani + Show 5 more

Open Access

https://doi.org/10.1021/acs.bioconjchem.0c00616

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Abstract

Cationic lipids (CLs) have gained significant attention among nonviral gene delivery vectors due to their ease of synthesis and functionalization with multivalent moieties. In particular, there is an increasing request for multifunctional CLs having gene delivery capacity and antibacterial activity. Herein, we describe the design and synthesis of a novel class of aminoglycoside (AG)-based multifunctional vectors with high transfection efficiency and noticeable antibacterial properties. Specifically, cationic amphiphiles were built on a triazine scaffold, allowing for an easy derivatization with up to three potentially different substituents, such as neomycin (Neo) that serves as the polar head and one or two lipophilic tails, namely stearyl (ST) and oleyl (OL) alkyl chains and cholesteryl (Chol) tail. With the aim to shed more light on the effect of different types and numbers of lipophilic moieties on the ability of CLs to condense and transfect cells, the performance of Neo–triazine-based derivatives as gene delivery vectors was evaluated and compared. The ability of Neo–triazine-based derivatives to act as antimicrobial agents was evaluated as well. Neo–triazine-based CLs invariably exhibited excellent DNA condensation ability, even at a low charge ratio (CR, +/−). Besides, each derivative showed very good transfection performance at its optimal CR on two different cell lines, along with negligible cytotoxicity. CLs bearing symmetric two-tailed OL proved to be the most effective in transfection. Interestingly, Neo–triazine-based derivatives, used as either free lipids or lipoplexes, exhibited strong antibacterial activity against Gram-negative bacteria, especially in the case of CLs bearing one or two aliphatic chains. Altogether, these results highlight the potential of Neo–triazine-based derivatives as effective multifunctional nonviral gene delivery vectors.

Highlights

In recent years, the concept of introducing foreign genetic material into cells has gained increasing attention within the clinical and scientific communities
Triazines have been poorly exploited for the synthesis of cationic lipids (CLs).[37−41] In the frame of a project dealing with the use of AG-based lipids and polymers as multifunctional gene delivery vehicles,[26,31,42] three types of Neo− triazine conjugates were synthesized through combinatorial chemistry, including monotailed, symmetric two-tailed, and asymmetric two-tailed Neo−triazine conjugates
Due to the harsh conditions needed for an effective third substitution (80 °C, 12 h) and the relatively low stability of AGs at high temperature, we introduced Neo at the end of the synthetic pathway by alkyne−azide “click” reaction with mono- or twotailed triazine intermediates bearing an N3−PEG2 arm