Synthesis and Biological Studies of New Temporin A Analogs Containing Unnatural Amino Acids in Position 7.

(1) Background: Antimicrobial resistance is growing at an extreme pace and has proven to be an urgent topic, for research into alternative treatments. Such a prospective possibility is hidden in antimicrobial peptides because of their low to no toxicity, effectiveness at low concentrations, and most importantly their ability to be used for multiple treatments. This work was focused on the study of the effect of the modification in position 7 of Temporin A on its biological activity; (2) Methods: The targeted peptides were synthesized using Fmoc/Ot-Bu SPPS. The antibacterial activity of the analogs was determined using the broth microdilution method and disk-diffusion method. In vitro tests were performed to determine the cytotoxicity, phototoxicity, and antiproliferative activity of the peptide analogs on a panel of tumor and normal cell lines; (3) Results: All analogs except DTCit showed good antibacterial activity, with DTDab having the best activity according to the disk-diffusion method. However, DTCit had an acceptable cytotoxicity, combined with good selectivity against the test MCF-7 cell line; (4) Conclusions: The obtained results revealed the importance of the basicity and length of the side chain at position 7 in the Temporin A sequence for both tested activities.

Effect of B2O3 on the Structure, Properties and Antibacterial Abilities of Sol-Gel-Derived TiO2/TeO2/B2O3 Powders.

This paper studies the influence of B2O3 on the structure, properties and antibacterial abilities of sol-gel-derived TiO2/TeO2/B2O3 powders. Titanium(IV) butoxide, telluric(VI) acid and boric acid were used as precursors. Differences were observed in the degree of decomposition of Ti butoxide in the presence of H3BO3 and H6TeO6 acids. The phase transformations of the obtained gels in the temperature range of 200-700 °C were investigated by XRD. Composite materials containing an amorphous phase and different crystalline phases (metallic Te, α-TeO2, anatase, rutile and TiTe3O8) were prepared. Heating at 400 °C indicated a crystalline-to-amorphous-phase ratio of approximately 3:1. The scanning electron microscopy (SEM) analysis showed the preparation of plate-like TiO2 nanoparticles. The IR results showed that the short-range order of the amorphous phases that are part of the composite materials consists of TiO6, BO3, BO4 and TeO4 structural units. Free B2O3 was not detected in the investigated compositions which could be related to the better connectivity between the building units as compared to binary TiO2/B2O3 compositions. The UV-Vis spectra of the investigated gels exhibited a red shift of the cut-off due to the presence of boron and tellurium units. The binary sample achieved the maximum photodegradation efficiency (94%) toward Malachite green dye under UV irradiation, whereas the ternary sample photoactivity was very low. The compositions exhibited promising antibacterial activity against E. coli NBIMCC K12 407.

Synthesis and Biological Studies on (KLAKLAK)2-NH2 Analog Containing Unnatural Amino Acid ß-Ala and Conjugates with Second Pharmacophore.

(1) Background: Peptides are good candidates for anticancer drugs due to their natural existence in the body and lack of secondary effects. (KLAKLAK)2 is an antimicrobial peptide that also shows good anticancer properties. (2) Methods: The Solid Phase Peptide Synthesis (Fmoc-strategy) was used for the synthesis of target molecules, analogs of (KLAKLAK)2-NH2. The purity of all compounds was monitored by HPLC, and their structures were proven using mass spectrometry. Cytotoxicity and antiproliferative effects were studied using 3T3 NRU and MTT tests, respectively. For determination of antimicrobial activity, the disc-diffusion method was used. Hydrolytic stability at three pH values, which mimic the physiological pH in the body, was investigated by means of the HPLC technique. (3) Results: A good selective index against MCF-7 tumor cell lines, combined with good cytotoxicity and antiproliferative properties, was revealed for conjugates NphtG-(KLAKLAK)2-NH2 and Caf-(KLAKLAK)2-NH2. The same compounds showed very good antifungal properties and complete hydrolytic stability for 72 h. The compound Caf-(KLß-AKLß-AK)2-NH2 containing ß-Ala in its structures exhibited good antimicrobial activity against Escherichia coli K12 407 and Bacillus subtilis 3562, in combination with very good antiproliferative and cytotoxic properties, as well as hydrolytic stability. (4) Conclusions: The obtained results reveal that all synthesized conjugates could be useful for medical practice as anticancer or antimicrobial agents.

The Influence of Nd and Sm on the Structure and Properties of Sol-Gel-Derived TiO2 Powders.

TiO2 nanopowders modified by Nd and Sm were prepared using the sol-gel technique. It was found by XRD analysis that the samples containing Sm are amorphous up to 300 °C, while those with Nd preserve a mixed organic-inorganic amorphous structure at higher temperatures (400 °C). The TiO2 (rutile) was not detected up to 700 °C in the presence of both modified oxides. TiO2 (anatase) crystals found at about 400 °C in the Sm-modified sample exhibited an average crystallite size of about 25-30 nm, while doping with Nd resulted in particles of a lower size-5-10 nm. It was established by DTA that organic decomposition is accompanied by significant weight loss occurring in the temperature range 240-350 °C. Photocatalytic tests showed that the samples heated at 500 °C possess photocatalytic activity under UV irradiation toward Malachite green organic dye. Selected compositions exhibited good antimicrobial activity against E. coli K12 and B. subtilis.

Synthesis, Antitumor and Antibacterial Studies of New Shortened Analogues of (KLAKLAK)2-NH2 and Their Conjugates Containing Unnatural Amino Acids.

(1) Background: (KLAKLAK)2 is a representative of the antimicrobial peptide group which also shows good anticancer properties. (2) Methods: Herein, we report synthesis using SPPS and characterization by HPLC/MS of a series of shortened analogues of (KLAKLAK)2. They contain single sequence KLAKLAK as C-terminal amides. In addition, substitution of some natural amino acids with unnatural ß-Ala and nor-Leu is realized. In addition, these structures are conjugated with second pharmacophore with well proven anticancer properties 1,8-naphthalimide or caffeic acid. Cytotoxicity, antiproliferative effect and antimicrobial activity of newly synthesized structures were studied. (3) Results: The obtained experimental results reveal significant selective index for substances with common chemical structure KLßAKLßAK-NH2. The antibacterial properties of newly synthesized analogues at two different concentrations 10 µM and 20 µM, were tested against Gram-negative microorganisms Escherichia coli K12 407. Only two of the studied compounds KLAKLAK-NH2 and the one conjugated with second pharmacophore 1,8-naphthalimide and unnatural amino acid nor-Leu showed moderate activity against tested strains at concentration of 20 µM. (4) Conclusions: The obtained results reveal that the introducing of 1,8-naphthalimideGly- and Caf- increase the cytotoxicity and antiproliferative activity of the peptides but not their selectivity. Only two compounds KLAKLAK-NH2 and 1,8-naphthalimideGKnLAKnLAK-NH2 show moderate activity against Escherichia coli K12 at low concentration of 20µM.

Functional characteristics of lactobacilli from traditional Bulgarian fermented milk products.

After oral administration, probiotic lactobacilli meet a number of protection systems in the human body, such as exposure to gastric, pancreatic, and small intestinal juices. Overcoming these detrimental barriers allows living bacteria to adhere to the intestinal epithelium and permanently colonize the gastrointestinal tract (GIT), providing health benefits to the host. Based on this, the transit tolerance of 25 candidate probiotic lactobacilli from katak, yoghurt, and white-brined and yellow cheese to simulated bile and small intestinal juices of variable pH was investigated. To establish their resistance, in vitro model systems based on modified MRS media and a longer duration of action (up to 24 h of incubation) were designed. Six of the strains studied were found to show strain-specific survival capacity with low viability in conditions simulating stomach acidity and high resistance to bile and intestinal juices. In addition, the adherence capability (autoaggregation and hydrophobicity) of the strains was determined. Obtained results allowed to select Lactobacillus strains with high survival ratios while passing through the GIT and good adherence properties, which make them suitable for the development of new probiotics.

Study of potential biomedical application of sol-gel derived Zn-doped SiO2-hydroxypropyl cellulose nanohybrids.

A series of Zn-doped hybrid materials based on silica from tetraethoxysilane (TEOS) and hydroxypropyl cellulose (HPC) were prepared by a sol-gel route. The structure, morphology and thermal behavior of synthesized hybrids were characterized by infrared (IR) spectroscopy, ultraviolet-visible spectroscopy (UV-Vis), transmission electron microscopy (TEM) and differential thermal analysis with thermogravimetric analysis (DTA/TG). The obtained materials were investigated for a potential biomedical application. The antibacterial properties of hybrids were investigated by measuring the inhibition zones formed around the materials containing different zinc content in presence of reference strains of Gram-positive and Gram-negative bacteria. The biocompatibility tests showed no cytotoxicity and genotoxicity, as well as no changes in actin cytoskeleton organization for hybrids with Zn content below 5 wt%.