Formation Of Noncovalent Complexes Research Articles

Enantioselective Reduction of Noncovalent Complexes of Amino Acids with CuII via Resonant Collision-Induced Dissociation: Collision Energy, Activation Duration Effects, and RRKM Modeling.

Formation of noncovalent complexes is one of the approaches to perform chiral analysis with mass spectrometry. Enantiomeric distinction of amino acids (AAs) based on the relative rate constants of competitive fragmentations of quaternary copper complexes is an efficient method for chiral differentiation. Here, we studied the complex [CuII,(Phe,PhG,Pro-H)]+ (m/z 493) under resonant collision-induced dissociation conditions while varying the activation time. The precursor ion can yield two main fragments through the loss of the non-natural AA phenylglycine (PhG): the expected product ion [CuII,(Phe,Pro-H)]+ (m/z 342) and the reduced product ion [CuI,(Phe,Pro)]+ (m/z 343). Enantioselective reduction describes the difference in relative abundance of these ions, which depends on the chirality of the precursor ion: the formation of the reduced ion m/z 343 is favored in homochiral complexes (DDD) compared to heterochiral complexes (such as LDD). Energy-resolved mass spectrometry data show that reduction, which arises from rearrangement, is favored at a low collision energy (CE) and long activation time (ActT), whereas direct cleavage preferentially occurs at a high CE and short ActT. These results were confirmed with kinetic modeling based on RRKM theory. For this modeling, it was necessary to set a pre-exponential factor as a reference, so that the E0 values obtained are relative values. Interestingly, these simulations showed that the critical energy E0 required to form the reduced ion is comparable in both homochiral and heterochiral complexes. However, the formation of product ion m/z 342 through direct cleavage is associated with a lower E0 in heterochiral complexes. Consequently, enantioselectivity would not be caused by enhanced reduction in homochiral complexes but rather by direct cleavage being favored in heterochiral complexes.

Effect of phlorotannins modification on the physicochemical, structural and functional properties of soybean protein isolate and controlled hydrolysates: Covalent and non-covalent interactions

In this research, the physicochemical, structural and functional properties of soybean protein isolate (SPI) as well as controlled hydrolysates (HSPI) were investigated by treatment with different concentrations of phlorotannins (PT) (0, 0.2, 0.5, 1.0, 2.0, mg/mL) through covalent and non-covalent interactions. Native/SDS-PAGE confirmed the formation of covalent and non-covalent complexes between PT and SPI/HSPI, and the presence of covalent bonds other than disulfide bonds in the covalent complexes (C–N, C–S bonds). After binding to PT, the secondary and tertiary structures of SPI/HSPI were changed. The secondary structure of complexes was transformed from β-sheet and β-turn to α-helix and random coil, and higher rates of transformation for covalent complexes. With the increase of PT concentration, the fluorescence and UV intensity of covalent/non-covalent complexes gradually reduced and rose, respectively; covalent complexes had lower fluorescence and higher UV intensity at equal concentrations. The particle size and ζ-potential of covalent/non-covalent complexes rose with PT concentration, and the PDI decreased and then rose, reaching a minimum value of 0.21 (HSPI-PT covalent) at PT of 0.5 mg/mL. TEM showed PT addition induced the aggregation of the complexes to increase. Meanwhile, surface hydrophobicity reduced while the antioxidant activity rose. Solubility, foaming activity and emulsifying activity of HSPI-PT covalent complexes first rose and then declined, and were optimal at PT of 0.5 mg/mL, with 78%, 250% and 97 m2/g, respectively. These will provide references for the improvement of soy protein properties and the application of complexes in formulated foods.

In Silico Evaluation of Oligomeric Representations for Molecularly Imprinted Polymer Modeling Using a Biological Template.

Molecularly imprinted polymers (MIPs) have significant relevance to analytical sensing due to their functionalized and template-specific structurally complementary cavities, providing increased sensibility and specificity for instrumental analyses, thereby enabling a wide variety of applications, especially for biological processes. Designing and developing MIPs entirely by experimental approaches are time-consuming and costly processes; thus, computational tools are used to assess some of the most critical parameters for imprinting, such as ligand screening. A typical practice is to model functional ligands as monomers; however, this representation fails to predict how ligand-template interactions evolve during polymer growth. In this context, this work aims to evaluate whether additional oligomeric representations affect the formation of noncovalent complexes between typical ligands and the P31 Asian lineage Zika virus epitope, using classical molecular dynamics. The ligands 2-vinylpyridine, 4-vinylaniline, acrylic acid, acrylamide, and 2-hidroxyethyl methacrylate were simulated as monomers, trimers, pentamers, and decamers, and their influence on the epitope structural conservation and ligand-template interactions were evaluated. Analyses of root-mean-square deviation, fluctuation, radius of gyration, pair correlation function, and number of hydrogen bonding-type interactions were conducted, showing the ligand chain size had an influence on the complex formation. However, this influence had no discernible pattern, exhibiting better performance in some cases while noninfluential in others. Of particular significance, in terms of epitope structural conservation, distinct oligomeric chains led to the selection of the distinct most interactive ligands. This observation raises important questions regarding the use of oligomeric chains in MIP simulations, thus prompting the need for further investigations of this subject.

Deciphering Structural Determinants Distinguishing Active from Inactive Cell-Penetrating Peptides for Cytosolic mRNA Delivery.

The formation of noncovalent complexes by mixing of positively charged polymers with negatively charged oligonucleotides (ONs) is a widely explored concept in nanomedicine to achieve cellular delivery of ONs. Uptake of ON complexes occurs through endocytosis, which then requires release of ON from endosomes. As one type of polymer, cell-penetrating peptides (CPPs) are being used which are peptides of about 8-30 amino acids in length. However, only a few CPPs yield effective cytosolic ON delivery and activity. Several strategies have been devised to increase cellular uptake and enhance endosomal release, among which an increase of osmotic pressure through the so-called proton sponge effect, disruption of membrane integrity through membrane activity, and disulfide-mediated polymerization. Here, we address the relevance of these concepts for mRNA delivery by incorporating structural features into the human lactoferrin-derived CPP, which shows uptake but not delivery. The incorporation of histidines was explored to address osmotic pressure and structural motifs of the delivery-active CPP PepFect14 (PF14) to address membrane disturbance, and finally, the impact of polymerization was explored. Whereas oligomerization increased the stability of polyplexes against heparin-induced decomplexation, neither this approach nor the incorporation of histidine residues to promote a proton-sponge effect yielded activity. Also, the replacement of arginine residues with lysine or ornithine residues, as in PF14, was without effect, even though all polyplexes showed cellular uptake. Ultimately, sufficient activity could only be achieved by transferring amphipathic sequence motifs from PF14 into the hLF context with some benefit of oligomerization demonstrating overarching principles of delivery for CPPs, lipid nanoparticles, and other types of delivery polymers.

Microfluidization of soybean protein isolate-tannic acid complex stabilized emulsions: Characterization of emulsion properties, stability and in vitro digestion properties

Emulsion stability and sustained-release can be improved with a non-covalent complexing of a soybean protein isolate (SPI) with -tannic acid (TA) and dynamic high-pressure microfluidization (DHPM). The microstructure, physicochemical properties, and interfacial properties were investigated. The properties of the DHPM-treated emulsions were improved significantly, with the 120 MPa DHPM-treated SPI-TA emulsion (SPI-TA 120) having the best microstructure. The highest interface protein content, viscosity and viscoelasticity at 120 MPa of pressure facilitated the stability of the emulsion. The oxidation kinetics of emulsions was established. It was demonstrated that the oxidation stability of SPI-TA 120 was higher than SPI and SPI-TA emulsions without DHPM treatment. In addition, DHPM-treated SPI-TA emulsions showed the most positive effect on the slow release of curcumin compared to the control group. The formation of non-covalent protein complexes with polyphenols and DHPM treatment effectively increases the stability of emulsions.

Mass spectrometry study of ascorbyl palmitate as an agent for nanosomes formation

Background. Study of properties and intermolecular interactions of biologically active compounds which can be used for the purposes of transmembrane drugs delivery is a topical task of modern molecular biophysics. Ascorbyl Palmitate (AP) as a fat-soluble form of vitamin C has recently attracted attention as a promising agent for formation of nanosomes for the “fat insoluble” drug molecules transfer through membranes. However, AP is not sufficiently characterized by up-to-date soft ionization mass spectrometric techniques. Objectives. The aim of the present work is to characterize AP and its intermolecular interactions by a number of mass spectrometric techniques: Electrospray Ionization (ESI), Laser Desorption/Ionization (LDI) and Matrix-Assisted Laser Desorption/Ionization (MALDI). The comparison of these techniques applicability to the study of AP intermolecular interactions as a drug delivery assisting agent is scheduled. Methods. ESI mass spectra are obtained with triple quadrupole Micromass Quattro mass spectrometer. LDI and MALDI experiments are performed by Autoflex II mass spectrometer. Results. In the ESI experiments in the positive ion mode abundant peaks of protonated and cationized AP molecules as well as the peaks of AP clusters nAP•H+ and nAP•Na+ (n=2÷4) are revealed in the mass spectra. This result testifies to the formation of stable noncovalent complexes of the AP molecules in the polar media and confirms the AP ability of formation nanosomes for drug delivery. Analysis of LDI and MALDI mass spectra of AP in positive and negative ion modes shows that in the presence of molecular ions of AP, the peaks of AP dimers or larger AP clusters are not recorded. The ESI probing of the model system containing AP and dipalmitoylphosphatidylcholine (DPPC) reveals stable AP•DPPC•H+ complex which models the AP intermolecular interactions with the phospholipid components of biomembranes and/or liposomes under AP functioning as a drug delivery assisting agent. Conclusions. The current study demonstrates the applicability of all tested mass spectrometric techniques for AP identification in solutions and solid phase, while for the purpose of examining of the AP noncovalent complexes formation and study of AP interactions with biomolecules the ESI is defined as the most effective technique.

Synergistic modification of structural and functional characteristics of whey protein isolate by soybean isoflavones non-covalent binding and succinylation treatment: A focus on emulsion stability

This investigation explored the structural and functional characteristics of whey protein isolate (WPI) and succinylated whey protein isolate (SAn-WPI) modified by varying concentrations (0.25, 0.5, 1.0, 2.0 mg/mL) of soybean isoflavones (SI) through non-covalent interactions. The results of SDS-PAGE and total phenolic contents confirmed the formation of non-covalent complexes. Molecular docking techniques, Fourier-transform infrared spectroscopy, and thermodynamic parameters obtained from fluorescence spectra revealed that non-covalent binding to SI primarily occurs through hydrophobic interactions and hydrogen bonding. Additionally, multispectroscopic techniques demonstrated that succinylation treatment resulted in partial unfolding of the proteins. This led to increased protein flexibility, exposing more binding sites and facilitating the binding between the proteins and SI. Therefore, SAn-WPI/SI exhibits stronger fluorescence quenching ability, antioxidant activity, and emulsifying property compared to WPI/SI at the same SI concentration. The findings illustrate the synergistic effect of non-covalent binding to SI and succinylation treatment on the WPI molecules, thereby promoting the utilization of protein-polyphenol complexes in food emulsion systems.

Noncovalent Host-Guest Complexes of Artemisinin with α-, β-, and γ- Cyclodextrin Examined by Structural Mass Spectrometry Strategies.

Cyclodextrins (CDs) are a family of macrocyclic oligosaccharides with amphiphilic properties, which can improve the stability, solubility, and bioavailability of therapeutic compounds. There has been growing interest in the advancement of efficient and reliable analytical methods that assist with elucidating CD host-guest drug complexation. In this study, we investigate the noncovalent ion complexes formed between naturally occurring dextrins (αCD, βCD, γCD, and maltohexaose) with the poorly water-soluble antimalarial drug, artemisinin, using a combination of ion mobility-mass spectrometry (IM-MS), tandem MS/MS, and theoretical modeling approaches. This study aims to determine if the drug can complex within the core dextrin cavity forming an inclusion complex or nonspecifically bind to the periphery of the dextrins. We explore the use of group I alkali earth metal additives to promote the formation of various noncovalent gas-phase ion complexes with different drug/dextrin stoichiometries (1:1, 1:2, 1:3, 1:4, and 2:1). Broad IM-MS collision cross section (CCS) mapping (n > 300) and power-law regression analysis were used to confirm the stoichiometric assignments. The 1:1 drug:αCD and drug:βCD complexes exhibited strong preferences for Li+ and Na+ charge carriers, whereas drug:γCD complexes preferred forming adducts with the larger alkali metals, K+, Rb+, and Cs+. Although the ion-measured CCS increased with cation size for the unbound artemisinin and CDs, the 1:1 drug:dextrin complexes exhibit near-identical CCS values regardless of the cation, suggesting these are inclusion complexes. Tandem MS/MS survival yield curves of the [artemisinin:βCD + X]+ ion (X = H, Li, Na, K) showed a decreased stability of the ion complex with increasing cation size. Empirical CCS measurements of the [artemisinin:βCD + Li]+ ion correlated with predicted CCS values from the low-energy theoretical structures of the drug incorporated within the βCD cavity, providing further evidence that gas-phase inclusion complexes are formed in these experiments. Taken together, this work demonstrates the utility of combining analytical information from IM-MS, MS/MS, and computational approaches in interpreting the presence of gas-phase inclusion phenomena.

Influence of environmental pH on the interaction properties of WP-EGCG non-covalent nanocomplexes.

Whey protein-epigallocatechin gallate (WP-EGCG) covalent conjugates and non-covalent nanocomplexes were prepared and compared using Fourier-transform infrared spectra. The effect of pH (at 2.6, 6.2, 7.1, and 8.2) on the non-covalent nanocomplexes' functional properties and the WP-EGCG interactions were investigated by studying antioxidant activity, emulsification, fluorescence quenching, and molecular docking, respectively. With the formation of non-covalent and covalent complexes, the amide band decreased; the -OH peak disappeared; the antioxidant activity of WP-EGCG non-covalent complexes was 2.59- and 2.61-times stronger than WP-EGCG covalent conjugates for 1-diphenyl-2-picryl-hydrazyl (DPPH) and ferric reducing ability of plasma(FRAP), respectively (particle size: 137 versus 370 nm). The antioxidant activity (DPPH 27.48-44.32%, FRAP 0.47-0.63) was stronger at pH 6.2-7.1 than at pH 2.6 and pH 8.2 (DPPH 19.50% and 26.36%, FRAP 0.39 and 0.41). Emulsification was highest (emulsifying activity index 181 m2 g-1 , emulsifying stability index 107%) at pH 7.1. The interaction between whey protein (WP) and EGCG was stronger under neutral and weakly acidic conditions: KSV (5.11-8.95 × 102 L mol-1 ) and Kq (5.11-8.95 × 1010 L mol s-1 ) at pH 6.2-7.1. Binding constants (pH 6.2 and pH 7.1) increased with increasing temperature. Molecular docking suggested that hydrophobic interactions played key roles at pH 6.2 and pH 7.1 (∆H > 0, ∆S > 0). Hydrogen bonding was the dominant force at pH 2.6 and pH 8.2 (∆H < 0, ∆S < 0). Environmental pH impacted the binding forces of WP-EGCG nanocomplexes. The interaction between WP and EGCG was stronger under neutral and weakly acidic conditions. Neutral and weakly acidic conditions are preferable for WP-EGCG non-covalent nanocomplex formation. © 2023 Society of Chemical Industry.

Curcumin Formulations for Better Bioavailability: What We Learned from Clinical Trials Thus Far?

Curcumin has been credited with a wide spectrum of pharmacological properties for the prevention and treatment of several chronic diseases such as arthritis, autoimmune diseases, cancer, cardiovascular diseases, diabetes, hemoglobinopathies, hypertension, infectious diseases, inflammation, metabolic syndrome, neurological diseases, obesity, and skin diseases. However, due to its weak solubility and bioavailability, it has limited potential as an oral medication. Numerous factors including low water solubility, poor intestinal permeability, instability at alkaline pH, and fast metabolism contribute to curcumin's limited oral bioavailability. In order to improve its oral bioavailability, different formulation techniques such as coadministration with piperine, incorporation into micelles, micro/nanoemulsions, nanoparticles, liposomes, solid dispersions, spray drying, and noncovalent complex formation with galactomannosides have been investigated with in vitro cell culture models, in vivo animal models, and humans. In the current study, we extensively reviewed clinical trials on various generations of curcumin formulations and their safety and efficacy in the treatment of many diseases. We also summarized the dose, duration, and mechanism of action of these formulations. We have also critically reviewed the advantages and limitations of each of these formulations compared to various placebo and/or available standard care therapies for these ailments. The highlighted integrative concept embodied in the development of next-generation formulations helps to minimize bioavailability and safety issues with least or no adverse side effects and the provisional new dimensions presented in this direction may add value in the prevention and cure of complex chronic diseases.

Mechanism of melanogenesis inhibition by Keggin-type polyoxometalates.

Abnormal melanin overproduction can result in hyperpigmentation syndrome in human skin diseases and enzymatic browning of fruits and vegetables. Recently, our group found that Keggin-type polyoxometalates (POMs) can efficiently inhibit tyrosinase activity. However, it remains unclear whether Keggin-type POMs exhibit optimal effects in vivo. Additionally, the inhibitory effect and mechanism of action of POMs on cellular tyrosinase activity and melanogenesis have been rarely reported. Here we demonstrate that our screened and synthesised PMo11Zn and GaMo12 show superior inhibitory effects on melanin formation as well as inhibition of cellular tyrosinase activity compared to other Keggin-type POMs. Intriguingly, we reveal that Keggin-type POMs competitively bind to tyrosinase mainly through more interactions with Cu2+ ions and the amino acid residue is capable of forming van der Waals, cation-π and hydrogen bonds, resulting in a reversible non-covalent complex formation. Our findings provide valuable insights into the design, synthesis and screening of polyoxometalates as multifunctional metallodrugs and food preservatives against hyperpigmentation.

Effective screening of Coulomb repulsions in water accelerates reactions of like-charged compounds by orders of magnitude

The reaction kinetics between like-charged compounds in water is extremely slow due to Coulomb repulsions. Here, we demonstrate that by screening these interactions and, in consequence, increasing the local concentration of reactants, we boost the reactions by many orders of magnitude. The reaction between negatively charged Coenzyme A molecules accelerates ~5 million-fold using cationic micelles. That is ~104 faster kinetics than in 0.5 M NaCl, although the salt is ~106 more concentrated. Rate enhancements are not limited to micelles, as evidenced by significant catalytic effects (104–105-fold) of other highly charged species such as oligomers and polymers. We generalize the observed phenomenon by analogously speeding up a non-covalent complex formation—DNA hybridization. A theoretical analysis shows that the acceleration is correlated to the catalysts’ surface charge density in both experimental systems and enables predicting and controlling reaction rates of like-charged compounds with counter-charged species.

Mechanistic study and synergistic effect on inhibition of α-amylase by structurally similar flavonoids

Natural plant-based flavonoids have the potential effect of reducing postprandial hyperglycaemia, thus they can be used to prevent and control type 2 diabetes mellitus. Here, to explore the antihyperglycemic mechanisms of flavonoids (apigenin, scutellarein, hispidulin and nepetin), their inhibition mechanisms against α-amylase were evaluated using multispectral methods and molecular docking in this study. As a result, nepetin, a competitive inhibitor, exhibited the best inhibitory effect than other three flavonoids, suggesting that adjacent dihydroxyl group on the B-ring played an important role in inhibiting the activity of α-amylase. At the same time, the combination of apigenin and scutellarein/hispidulin/nepetin synergistically improved their inhibitory effects on α-amylase. In addition, fluorescence quenching analysis indicated that the addition of flavonoids could result in the static quenching of α-amylase and the formation of non-covalent complexes. The interaction further led to the alteration of microenvironment and secondary structure of α-amylase, thus reducing the catalytic activity of enzyme. In conclusion, this finding can provide vital insights into the use of flavonoids, especially nepetin, as the potential drug candidates for the treatment of diabetes.

Tunning of Optoelectronic Properties in Nanohybrids of Peptide-Appended Perylenebisimides and Carbon Nanodots

Two hybrid materials based on carbon dots (C-dots) and two perylenebisimide (PBI)-linked peptides with different functionality have been fabricated. These hybrids are designed to facilitate the electronic interaction between the electron donor C-dots and acceptor PBI-appended peptides, and the optoelectronic properties of these nanohybrids are compared. Amazingly, it is observed that the C-dots have exhibited favorable interactions with the PBI molecule (P1) bearing a similar charge (both negative) and have not shown any electronic interaction with the PBI molecule having opposite charges at their termini (P2). The pH of the medium has been found to be the limiting factor that has inhibited the electronic interaction between C-dots and P2. The interaction between C-dots and P1 has been attributed to the noncovalent complex formation. UV–visible and fluorescence spectroscopic study, field emission gun transmission electron microscopy, and time-correlated single-photon counting studies are made for this purpose. This nanohybrid material shows the emergence of good photoswitching behavior with the highest photocurrent gain of 5.14 (Ion/Ioff) compared to that of only P1 or C-dots. The increase in the photoswitching property is attributed to efficient charge (electron–hole) separation and holds a future promise for fabricating a new nanohybrid material with a high photocurrent conversion efficiency.

Formation of soybean protein isolate-hawthorn flavonoids non-covalent complexes: Linking the physicochemical properties and emulsifying properties

In recent years, more and more attention had been paid to the combination of proteins and flavonoids, and several flavonoids had been reported to improve the physicochemical and emulsifying properties of proteins. This study investigated the effects of ultrasonic treatment (450 W for 10 min, 20 min, and 30 min) on the physicochemical properties, antioxidant activity, and emulsifying properties of soy protein isolate (SPI) -hawthorn flavonoids (HF) non-covalent complexes. The results showed that the addition of HF to SPI and 20 min of ultrasound could reduce α-helix and random coil, increase β-sheet and β-turn, and enhance fluorescence quenching. In addition, it decreased the particle size, zeta potential, surface hydrophobicity, and turbidity to 88.43 or 95.27 nm, −28.80 mV, 1250.42, and 0.23, respectively. The protein solubility, free sulfhydryl group, antioxidant activity, emulsifying activity index, and emulsifying stability index all increased to 73.93%, 15.07 μmol/g, 71.00 or 41.91%, 9.81 m2/g, and 67.71%, respectively. Moreover, high-density small and low-flocculation droplets were formed. Therefore, the combined ultrasound treatment and addition of HF to SPI is a more effective method for protein modification compared to ultrasound treatment alone. It provides a theoretical basis for protein processing and application in the future.

Formation and characterization of noncovalent ternary complexes based on whey protein concentrate, high methoxyl pectin, and phenolic acid

Protein-polysaccharide-polyphenol noncovalent ternary complexes possess unique physicochemical, structural, and functional properties. In the present study, ternary complexes based on whey protein concentrate (WPC; 2%, wt/vol) and high methoxyl pectin (HMP; 0.5%, wt/vol) complexes and 0.2 to 0.6% (wt/vol) chlorogenic acid (CA) or rosmarinic acid (RA) were formed and characterized at 3 pH values (4, 4.5, and 5). The pH conditions were decided according to phase diagram of WPC and HMP during acidification. Fluorescence quenching experiments indicated that WPC-HMP complexes bound RA stronger than CA and the binding constant increased with increasing pH for both phenolic acids. Particle size of ternary complexes decreased and absolute ζ-potential increased with pH values changing from 4 to 5, and RA influenced the particle size of WPC-HMP complexes greater than CA. The CA and RA in ternary complexes showed good stability against UV light with pH order of pH 5 > pH 4.5 > pH 4. Fourier-transform infrared spectroscopy spectra indicated the involvement of hydrogen bonding between WPC-HMP and CA or RA. Antibacterial tests showed that ternary complexes had good antibacterial activity against Staphylococcus aureus and Escherichia coli at concentrations of 6.2 mg/mL and the ability increased with decreasing pH values. All ternary complexes possessed strong scavenging radical capacities with median inhibitory concentration (IC50) values ranging from 2.71 ± 0.05 to 6.20 ± 0.41 μg/mL. Antioxidative ability of ternary complexes increased as pH went up and WPC-HMP-RA showed significantly higher antioxidative property compared with WPC-HMP-CA. Data may provide useful information for rational design of ternary complexes and applications of the formed complexes in food matrices such as beverages and emulsions.

DFT-D4 Insight into the Inclusion of Amphetamine and Methamphetamine in Cucurbit[7]uril: Energetic, Structural and Biosensing Properties.

The host–guest interactions of cucurbit[7]uril (CB[7]) as host and amphetamine (AMP), methamphetamine (MET) and their enantiomeric forms (S-form and R-form) as guests were computationally investigated using density functional theory calculations with the recent D4 atomic-charge dependent dispersion corrections. The analysis of energetic, structural and electronic properties with the aid of frontier molecular orbital analysis, charge decomposition analysis (CDA), extended charge decomposition analysis (ECDA) and independent gradient model (IGM) approach allowed to characterize the host–guest interactions in the studied systems. Energetic results indicate the formation of stable non-covalent complexes where R-AMP@CB[7] and S-AMP@CB[7] are more stable thermodynamically than R-MET@CB[7] and S-MET@CB[7] in gas phase while the reverse is true in water solvent. Based on structural analysis, a recognition mechanism is proposed, which suggests that the synergistic effect of van der Waals forces, ion–dipole interactions, intermolecular charge transfer interactions and intermolecular hydrogen bonding is responsible for the stabilization of the complexes. The geometries of the complexes obtained theoretically are in good agreement with the X-ray experimental structures and indicate that the phenyl ring of amphetamine and methamphetamine is deeply buried into the cavity of CB[7] through hydrophobic interactions while the ammonium group remains outside the cavity to establish hydrogen bonds with the portal oxygen atoms of CB[7].

Chemistry of Peptide-Oligonucleotide Conjugates: A Review.

Peptide-oligonucleotide conjugates (POCs) represent one of the increasingly successful albeit costly approaches to increasing the cellular uptake, tissue delivery, bioavailability, and, thus, overall efficiency of therapeutic nucleic acids, such as, antisense oligonucleotides and small interfering RNAs. This review puts the subject of chemical synthesis of POCs into the wider context of therapeutic oligonucleotides and the problem of nucleic acid drug delivery, cell-penetrating peptide structural types, the mechanisms of their intracellular transport, and the ways of application, which include the formation of non-covalent complexes with oligonucleotides (peptide additives) or covalent conjugation. The main strategies for the synthesis of POCs are viewed in detail, which are conceptually divided into (a) the stepwise solid-phase synthesis approach and (b) post-synthetic conjugation either in solution or on the solid phase, especially by means of various click chemistries. The relative advantages and disadvantages of both strategies are discussed and compared.

Photonics of meso-substituted carbocyanine dyes in solutions and in complexes with DNA

Spectral-fluorescent and photochemical properties (photoisomerization and generation of the triplet state) of meso-substituted cationic carbocyanine dyes, 3,3′-di-(β-hydroxyethyl)-5,5′-dimethoxy-9-ethylthiacarbocyanine iodide (K1) and 3,3′-di-(β-hydroxyethyl)-9-methylthiacarbocyanine iodide (K2), have been studied in solutions and in the presence of DNA. In solutions, on passing from acetonitrile to dioxane, a growth of fluorescence of the dyes is observed due to a shift of the equilibrium of cis/trans isomers toward the fluorescent trans-isomer. Upon flash photolysis of dye solutions in dioxane, the formation and subsequent decay of the cis-photoisomers of the dyes are observed.In aqueous solutions, the interaction with DNA leads to the formation of noncovalent complexes of K1 and K2 with DNA, which is accompanied by a significant increase in the fluorescence intensity. The results of the molecular docking experiments showed the possibility of several types of binding, which was confirmed by the data obtained from other experiments. The effects of temperature and additions of NaCl on the stability of the dye–DNA complexes were studied. The spectral-fluorescent data were used to estimate the binding constants of the dyes with DNA and other characteristics of the dyes that are important for their use as probes.Upon flash photolysis of the dyes in complexes with DNA, photoisomerization is not observed, but the quantum yield of intersystem crossing to the triplet state increases. The decay of the triplet states occurs by a two-exponential law. The rate constants for quenching of the triplet states of the dyes complexed with DNA by oxygen were found to be lower than the expected values for diffusion-controlled quenching (taking into account the spin statistical factor 1/9), which is explained by the steric factor of complexation.

Observation of Intact and Proteolytically Cleaved Amyloid-Beta (1-40)-Oleuropein Noncovalent Complex at Neutral pH by Mass Spectrometry.

Mass spectrometry analyses carried out on mass spectrometers equipped with soft ionization sources demonstrated their utility in the assessment of the formation of noncovalent complexes and the localization of the binding sites. Direct analyses by mass spectrometry of the noncovalent complex formed in acidic and mildly acidic environments by amyloid beta (1–40) peptide and oleuropein have been previously described, and, in several studies, the absorption, metabolism, excretion, and the implications in the prevention and therapy of Alzheimer’s disease of oleuropein have been investigated. Our paper presents modifications of the method previously employed for noncovalent complex observation, namely, the amyloid beta (1–40) pretreatment, followed by an increase in the pH and replacement of the chemical environment from ammonium acetate to ammonium bicarbonate. The formation of noncovalent complexes with one or two molecules of oleuropein was detected in all chemical solutions used, and the amyloid beta (17–28) binding site was identified via proteolytic experiments using trypsin prior to and after noncovalent complex formation. Our results highlight the importance of further studies on the effect of oleuropein against amyloid beta aggregation.