Self-Assembly of Highly Stable Nanoparticles by Amphiphilic Glycolurils for Efficient Intracellular Short DNA Delivery

doi:10.6023/cjoc202501022

Abstract

Four glycoluril-based amphiphilic molecular clips (AMCs) M1~M4 have been prepared for intracellular delivery of short DNA. M1~M4 have two methyl groups on its convex surface and four cations on its aromatic side arm, which can be used to construct self-assembled nanoparticles in aqueous solution driven by hydrophobic interaction. Dynamic light scattering experiments show that M1 and M2 can be driven hydrophobically to aggregate into extremely stable nanoparticles in water at the micromolar concentrations. Fluorescence titration and zeta potential experiments support that the nanoparticles formed by M1 and M2 are able to efficiently encapsulate short DNA (sDNA). Fluorescence imaging and flow cytometry studies reveal that their nano sizes enable intracellular delivery of the encapsulated sDNA into both normal and cancer cells, with delivery percentage reaching up to 94%, while in vitro experiments indicate that the two compounds have excellent biocompatibility and low cytotoxicity.

Key words: self-assembly, glycoluril, amphiphilicity, nanoparticle, DNA delivery

Cite this article

Congying Guo, Rui Gao, Qian Li, Hui Wang, Danwei Zhang, Wei Zhou, Zhan-Ting Li. Self-Assembly of Highly Stable Nanoparticles by Amphiphilic Glycolurils for Efficient Intracellular Short DNA Delivery[J]. Chinese Journal of Organic Chemistry, 2025, 45(8): 2945-2952.

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Fig. & Tab. 7

Scheme 1. Compounds M1~M4 and the routes for their preparations

Figure 1. DLS profile of the aqueous solutions of compounds (a) M1 and (b) M2

Figure 2. Tentative mechanism for the formation of nanoparticles by M1~M4 through hydrophobically driven aggregation of the glycoluril unit and their delivery for short DNA

Figure 3. Fluorescence spectra of Cy5-ssDNA-21 (0.25 μmol/L, λex=625 nm) in water with the addition of (a) M1 and (b) M2 (all the experiments were conducted at 25 ℃)

Figure 4. Zeta potentials of the solutions of (a) M1, M1＋Cy5-ssDNA-21 (10 μmol/L), and (b) M2, M2＋Cy5-ssDNA-21 (10 μmol/L) (all the experiments were conducted at 25 ℃)

Figure 5. CLSM images of MCF-7/ADR cells after incubation with Cy5-ssDNA-21, Cy5-ssDNA-21＋M4, Cy5-ssDNA-21＋M3, Cy5-ssDNA-21＋M2, Cy5-ssDNA-21＋M1 and Cy5- ssDNA-21＋Lipo2000 for 2 h ([Cy5-ssDNA-21]=2.5 μg/mL, [M4/M3/M2/M1/Lipo2000]=20.0 μg/mL)

Figure 6. (a) Delivery (internalization) of Cy5-ssDNA-21 (2.5 μg/mL) and Cy5-dsDNA-21 (5 μg/mL) into MCF-7/ADR cell line by M1~M4 (20 μg/mL for ssDNA and 40 μg/mL for dsDNA) and Lipo2000 (20 μg/mL for ssDNA and 40 μg/mL for dsDNA) after incubation in complete 1640 medium for 2 h, and (b) cell viability values (%) of MCF-7/ADR cell line evaluated by CCK-8 proliferation tests versus the incubation concentration of M1~M4 (the cells (≈2×104 per well) were incubated with M1~M4 at 37 ℃ for 24 h. Error bars represent the s.d. of uncertainty for each point)

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