Gold Nanosystems Covered with Doxorubicin/DNA Complexes: A Therapeutic Target for Prostate and Liver Cancer

Different gold nanosystems covered with DNA and doxorubicin (Doxo) were designed and synthesized for cancer therapy, starting from Au@16-Ph-16 cationic nanoparticles and DNA–Doxo complexes prepared under saturation conditions. For the preparation of stable, biocompatible, and small-sized compacted Au@16-Ph-16/DNA–Doxo nanotransporters, the conditions for the DNA–Doxo compaction process induced by gold nanoparticles were first explored using fluorescence spectroscopy, circular dichroism and atomic force microscopy techniques. The reverse process, which is fundamental for Doxo liberation at the site of action, was found to occur at higher CAu@16-Ph-16 concentrations using these techniques. Zeta potential, dynamic light scattering and UV–visible spectroscopy reveal that the prepared compacted nanosystems are stable, highly charged and of adequate size for the effective delivery of Doxo to the cell. This fact is verified by in vitro biocompatibility and internalization studies using two prostate cancer-derived cell lines (LNCaP and DU145) and one hepatocellular carcinoma-derived cell line (SNU-387), as well as a non-tumor prostate (PNT2) cell line and a non-hepatocarcinoma hepatoblastoma cell line (Hep-G2) model used as a control in liver cells. However, the most outstanding results of this work are derived from the use of the CI+NI combined treatments which present strong action in cancer-derived cell lines, while a protective effect is observed in non-tumor cell lines. Hence, novel therapeutic targets based on gold nanoparticles denote high selectivity compared to conventional treatment based on free Doxo at the same concentration. The results obtained show the viability of both the proposed methodology for internalization of compacted nanocomplexes inside the cell and the effectiveness of the possible treatment and minimization of side effects in prostate and liver cancer

Sodium Lauryl Sulfate-Conjugated Cationic Gemini-Surfactant-Capped Gold Nanoparticles as Model System for Biomolecule Recognition

Surfactant-based nanostructures are promising materials for designing novel colorimetric biosensors based on aggregation/disaggregation phenomena. In this work, a colorimetric sensor based on the plasmonic shift of surfactant-capped gold nanoparticles via the disaggregation mechanism was developed. To perform this, the optimum SDS concentration was firstly determined in order to form Au@16-s-16/SDS complex aggregates with a well-defined SPR band in the blue region. Once the optimal SDS concentration for Au@16-s-16 aggregation was established, the sensing method depended on the nature of the electrostatic charge of the biopolymer studied where both the strength of the biopolymer/SDS and biopolymer/Au@16-s-16 interactions and the cationic gold nanoparticles play a key role in the disaggregation processes. As a result, an instantaneous color change from blue to red was gradually observed with increasing biopolymer concentrations. The response of the sensor was immediate, avoiding problems derived from time lapse, and highly dependent on the order of addition of the reagents, with a detection limit in the nanomolar and picomolar range for DNA and Lysozyme sensing, respectively. This behavior can be correlated with the formation of different highly stabilized Au@16-s-16/biopolymer/SDS complexes, in which the particular biopolymer conformation enhances the distance between Au@16-s-16 nanoparticles among the complexes.Junta de Andalucía 2021/FQM-386Universidad de Sevilla 2021/00001297; 2022/0000027

DNA conformational changes induced by cationic gemini surfactants: the key to switching DNA compact structures into elongated forms

The DNA conformational changes induced by different members of the N,N0-bis(dimethyldodecyl)-a-ualkanediammonium dibromide series (m-s-m, m ¼ 12, s ¼ 3 and 6) and the analogous series of hexadecyl gemini surfactants (m ¼ 16, s ¼ 3 and 6) were investigated in aqueous media by means of circular dichroism (CD), zeta potential, dynamic light scattering (DLS), viscometry, and atomic force microscopy (AFM) methods. The measurements were carried out by varying the gemini surfactant–DNA molar ratio, R ¼ Cm-s-m/CDNA. For the conditions investigated two significantly different conformational changes were observed, the second of them being worth noting. At low molar ratios, all methods concurred by showing that gemini surfactants were able to form ordered aggregates which precedes DNA compaction. The second effect observed, at high molar ratios, corresponds to the transition from the compact state to a new more extended conformation. The degree of decompaction and the morphologies of the visualized structures are different not only depending on the surfactant tail's length, but also on the spacer's length. The results obtained for the 16-3-16/DNA and 16-6-16/DNA systems point out that the compaction/decompaction processes are somewhat different to those previously visualized for the analogous monoquaternary chain surfactant CTAB.Consejería de Educación y Ciencia of the Junta de Andalucía FQM-0362

Use of Nanoparticles to Prevent Resistance to Antibiotics—Synthesis and Characterization of Gold Nanosystems Based on Tetracycline

Antimicrobial resistance (AMR) is a serious public health problem worldwide which, according to the World Health Organization (WHO), requires research into new and more effective drugs. In this work, both gold nanoparticles covered with 16-3-16 cationic gemini surfactant (Au@16-3-16) and DNA/tetracycline (DNA/TC) intercalated complexes were prepared to effectively transport tetracycline (TC). Synthesis of the Au@16-3-16 precursor was carried out by using trihydrated gold, adding sodium borohydride as a reducing agent and the gemini surfactant 16-3-16 as stabilizing agent. Circular dichroism and atomic force microscopy techniques were then used to ascertain the optimal R range of the relationship between the concentrations of Au@16-3-16 and the DNA/TC complex (R = CAu@16-3-16/CDNA) that allow the obtainment of stable and compact nanosystems, these characteristics being fundamental for their use as antibiotic transporters. Stability studies over time were carried out for distinct selected Au@16-3-16 and Au@16-3-16/DNA-TC nanoformulations using the ultraviolet–visible spectrophotometry technique, checking their stability for at least one month. In addition, in order to know the charge and size distribution of the nanocomplexes, DLS and zeta potential measurements were performed in the solution. The results showed that the characterized nanosystems were highly charged, stable and of a reduced size (<100 nm) that allows them to cross bacterial membranes effectively (>1 µm). Once the different physicochemical characteristics of the gold nanosystems were measured, Au@16-3-16 and Au@16-3-16/DNA-TC were tested on Escherichia coli and Staphylococcus aureus to study their antibacterial properties and internalization capacity in microbes. Differences in the interaction of the precursors and the compacted nanosystems generated were observed in Gram-positive and Gram-negative bacteria, possibly due to membrane damage or electrostatic interaction with internalization by endocytosis. In the internalization experiments, depending on the treatment application time, the greatest bacterial destruction was observed for all nanoformulations explored at 18 h of incubation. Importantly, the results obtained demonstrate that both new nanosystems based on TC and Au@16-3-16 precursors have optimal antimicrobial properties and would be beneficial for use in patients, avoiding possible side effects.Junta de Andalucía FQM-386Universidad Pablo de Olavide 00001297Universidad de Sevilla 0000027

Synergistic Antibacterial Effects of Amoxicillin and Gold Nanoparticles: A Therapeutic Option to Combat Antibiotic Resistance

Compacted Au@16-mph-16/DNA-AMOX (NSi) nanosystems were prepared from amoxicillin (AMOX) and precursor Au@16-mph-16 gold nanoparticles (Ni) using a Deoxyribonucleic acid (DNA) biopolymer as a glue. The synthesized nanocarrier was tested on different bacterial strains of Escherichia coli, Staphylococcus aureus, and Streptococcus pneumoniae to evaluate its effectiveness as an antibiotic as well as its internalization. Synthesis of the nanosystems required previous structural and thermodynamic studies using circular dichroism (CD) and UV-visible techniques to guarantee optimal complex formation and maximal DNA compaction, characteristics which facilitate the correct uptake of the nanocarrier. Two nanocomplexes with different compositions and structures, denoted NS1 and NS2, were prepared, the first involving external Au@16-mph-16 binding and the second partial intercalation. The Ni and NSi nanosystems obtained were characterized via transmission electron microscopy (TEM), zeta potential, and dynamic light scattering (DLS) techniques to measure their charge, aggregation state and hydrodynamic size, and to verify their presence inside the bacteria. From these studies, it was concluded that the zeta potential values for gold nanoparticles, NS1, and NS2 nanosystems were 67.8, −36.7, and −45.1 mV. Moreover, the particle size distribution of the Au@16-mph-16 gold nanoparticles and NS2 nanoformulation was found to be 2.6 nm and 69.0 nm, respectively. However, for NS1 nanoformulation, a bimodal size distribution of 44 nm (95.5%) and 205 nm (4.5%) was found. Minimal inhibitory concentration (MIC) values were determined for the bacteria studied using a microdilution plates assay. The effect on Escherichia coli bacteria was notable, with MIC values of 17 µM for both the NS1 and NS2 nanosystems. The Staphylococcus aureus chart shows a greater inhibition effect of NS2 and NP2 in non-diluted wells, and clearly reveals a great effect on Streptococcus pneumoniae, reaching MIC values of 0.53 µM in more diluted wells. These results are in good agreement with TEM internalization studies of bacteria that reveal significant internalization and damage in Streptococcus pneumoniae. In all the treatments carried out, the antibiotic capacity of gold nanosystems as enhancers of amoxicillin was demonstrated, causing both the precursors and the nanosystems to act very quickly, and thus favoring microbial death with a small amount of antibiotic. Therefore, these gold nanosystems may constitute an effective therapy to combat resistance to antibiotics, in addition to avoiding the secondary effects derived from the administration of high doses of antibiotics.Junta de Andalucía 2021/FQM-386Universidad de Sevilla 2021/00001297, 2022/00000274, 2023/0000030

On the rearrangement of N-aryl-N-Boc-phosphoramidates to N-Boc-protected o-aminoarylphosphonates

Various arylamines were converted in two steps to N-Boc-N-arylphosphoramidates. LiTMP and LDA induced directed ortho-metalation at temperatures from −78 to 0 °C. The ensuing [1,3]-migration of the phosphorus atom with its substituents from the nitrogen to the ortho-carbanionic carbon atom gave N-Boc-protected o-aminoarylphosphonates. The nature of the substituent of 3-substituted phenylphosphoramidates strongly influenced the regioselectivity of phosphonate formation. A crossover experiment with a deuterated phosphoramidate proved the intramolecular course of the rearrangement. Three representative N-Boc-o-aminoarylphosphonates were deprotected to access the corresponding o-aminoarylphosphonic acids.© The Author(s) 201

Biocompatible DNA/5-fluorouracil-gemini surfactant-functionalized gold nanoparticles as promising vectors in lung cancer therapy

The design and preparation of novel nanocarriers to transport cancer drugs for chemotherapy purposes is an important line of research in the medical field. A new 5-fluorouracil (5-Fu) transporter was designed based on the use of two new biocompatible gold nanosystems: (i) a gold nanoparticle precursor, Au@16-Ph-16, stabilized with the positively charged gemini surfactant 16- Ph-16, and (ii) the compacted nanocomplexes formed by the precursor and DNA/5-Fu complexes, Au@16-Ph-16/DNA-5-Fu. The physicochemical properties of the obtained nanosystems were studied by using UV-visible spectroscopy, TEM, dynamic light scattering, and zeta potential techniques. Method tuning also requires the use of circular dichroism, atomic force microscopy, and fluorescence spectroscopy techniques for the prior selection of the optimal relative Au@16-Ph-16 and DNA concentrations (R = CAu@16-Ph-16/CDNA), biopolymer compaction/decompaction, and 5-Fu release from the DNA/5-Fu complex. TEM experiments revealed the effective internalization of the both precursor and Au@16-Ph-16/DNA-5-Fu-compacted nanosystems into the cells. Moreover, cytotoxicity assays and internalization experiments using TEM and confocal microscopy showed that the new strategy for 5-Fu administration enhanced efficacy, biocompatibility and selectivity against lung cancer cells. The differential uptake among different formulations is discussed in terms of the physicochemical properties of the nanosystems.Universidad de Sevilla 2019/00000570, 2020/00001068, 2010/00000762, PP2016-5937, 2020/00001073Junta de Andalucía 1804032996/2017, 2018/00000810, 2019/FQM-38