In Vitro and In Silico Studies on Cytotoxic Properties of Oxythiamine and 2'-Methylthiamine.

It is important to search for cytostatic compounds in order to fight cancer. One of them could be 2'-methylthiamine, which is a thiamine antimetabolite with an additional methyl group at the C-2 carbon of thiazole. So far, the cytostatic potential of 2'-methylthiamine has not been studied. We have come forward with a simplified method of synthesis using commercially available substrates and presented a comparison of its effects, as boosted by oxythiamine, on normal skin fibroblasts and HeLa cancer cells, having adopted in vitro culture techniques. Oxythiamine has been found to inhibit the growth and metabolism of cancer cells significantly better than 2'-methylthiamine (GI50 36 and 107 µM, respectively), while 2'-methylthiamine is more selective for cancer cells than oxythiamine (SI = 180 and 153, respectively). Docking analyses have revealed that 2'-methylthiamine (ΔG -8.2 kcal/mol) demonstrates a better affinity with thiamine pyrophosphokinase than thiamine (ΔG -7.5 kcal/mol ) and oxythiamine (ΔG -7.0 kcal/mol), which includes 2'-methylthiamine as a potential cytostatic. Our results suggest that the limited effect of 2'-methylthiamine on HeLa arises from the related arduous transport as compared to oxythiamine. Given that 2'-methylthiamine may possibly inhibit thiamine pyrophosphokinase, it could once again be considered a potential cytostatic. Thus, research should be carried out in order to find the best way to improve the transport of 2'-methylthiamine into cells, which may trigger its cytostatic properties.

Polymer Network Comprised of Steroidal Rotors as a Promising Storage Material.

The front cover artwork is provided by Dr habil. Izabella Jastrzebska's group from the University of Bialystok, Poland. The image shows a polymeric network with molecular rotors (MR) as crosslinks. The MR rotation is slowed or inhibited when a molecule of stored gas is placed inside the polymer material. Read the full text of the Research Article at 10.1002/cphc.202300793.

Molecular Dynamics of Steroidal Rotors Probed by Theoretical, Spectroscopic and Dielectric Methods.

Our study focuses on molecular rotors with fast-moving rotators and their potential applications in the development of new amphidynamic crystals. Steroidal molecular rotors with a dipolar fluorine-substituted phenyl group as the rotator were synthesized and characterized. Three different rotors were investigated with varying numbers of fluorine atoms. A comprehensive analysis was performed using vibrational spectroscopy (Raman, FT-IR), electronic circular dichroism (ECD), and dielectric response to understand the behavior of the investigated model rotors. The results were supported by theoretical calculations using Density Functional Theory (DFT) methods. The angle-dependent polarized Raman spectra confirmed the crystallinity of the samples. Nearly frequency and temperature-independent permittivity suggest low-frequency librational motion of stators. An in-depth analysis of ECD spectra revealed high conformational flexibility in solution, resulting in low ECD effects, while in the solid-state with restricted rotation, significant ECD effects were observed. These findings shed light on the conformational behavior and potential applications of the studied steroidal molecular rotors.

Polymer network comprised of steroidal rotors as promising storage material.

In this paper, we report a new generation of polymeric networks as potential functional material based on changes in molecular dynamics in the solid state. The material is obtained by free radical polymerization of a diacrylate derivative bearing a steroid (stator) and a 1,4-diethynyl-phenylene-d4 fragment (rotator). Polymer research using the PALS technique complements the knowledge about nanostructural changes occurring in the system in the temperature range -115 °C - +190 °C. It indicates the presence of two types of free nanovolumes in the system and the occurrence of phase transitions. The polymer is characterized using 1 H NMR, 2 H Solid Echo NMR, ATR-FTIR and Raman spectroscopies, thermal analysis, and porosimetry. It is proved that the applied procedure leads to the formation of a novel porous organic material containing multiple molecular rotors.

New Steroidal Selenides as Proapoptotic Factors.

Cytostatic and pro-apoptotic effects of selenium steroid derivatives against HeLa cells were determined. The highest cytostatic activity was shown by derivative 4 (GI50 25.0 µM, almost complete growth inhibition after three days of culture, and over 97% of apoptotic and dead cells at 200 µM). The results of our study (cell number measurements, apoptosis profile, relative expression of apoptosis-related APAF1, BID, and mevalonate pathway-involved HMGCR, SQLE, CYP51A1, and PDHB genes, and computational chemistry data) support the hypothesis that tested selenosteroids induce the extrinsic pathway of apoptosis by affecting the cell membrane as cholesterol antimetabolites. An additional mechanism of action is possible through a direct action of derivative 4 to inhibit PDHB expression in a way similar to steroid hormones.

Metal-promoted synthesis of steroidal ethynyl selenides having anticancer activity.

In this study, we have described simple and efficient methodology for the metal-promoted (Cu2I2) preparation of steroidal ethynyl selenides. The compounds were characterized using 1H, 13C and 77Se NMR, FT IR spectroscopy, and MS analysis. A proposed mechanism of the metal-promoted reaction involves the formation of a σ-bound copper acetylide. Due to the fact that organoselenium-based compounds possess a pleiotropic properties and associated with their promising biological activities, in the next step of the study biocompatibility and anticancer activity of the synthesized compounds was evaluated. Steroidal selenides were tested in vitro against estrogen-depend breast cancer cells MCF-7 using spectrophotometric, fluorometric and luminometric methods. Designed selenides showed high hemocompatibility, lack of toxicity against cardiomyocytes cell and great anti-cancer activity in vitro against estrogen-depend breast cancer cells upon 24 h of treatment. We revealed that selenides decrease the viability and proliferation ability of MCF-7 cells by induction of cell apoptosis. It has been noted that the overproduction of reactive oxygen species (ROS) and associated with its activation of Caspase 3/7 are a major mechanism that is responsible of selenides-caused cell death. These data indicate that organoselenium based compounds have great antineoplastic potential and might be developed as novel class of agents dedicated to the breast-cancer therapies.

Simple Zn-Mediated Seleno- and Thio-Functionalization of Steroids at C-1 Position.

Here we report the reaction in the biphasic system of the in situ prepared selenols and thiols with 1,4-androstadiene-3,17-dione (1) or prednisone acetate (2) having α,ß-unsaturated ketone as an electrophilic functionalization. The Michael-type addition reaction resulted to be chemo- and stereoselective, affording a series of novel steroidal selenides and sulfides. This is an example of a one-step, eco-friendly process that bypasses some of the main concerns connected with the bad smell and the toxicity of these seleno- and thio-reagents. Furthermore, we demonstrated that the proposed methodology offers the possibility to prepare libraries of steroids variously and selectively decorated with different organochalcogen moieties at the C1 position starting from 1,4-androstadienic skeletons and leaving unaltered the C4-C5 unsaturation. Based on the data reported in the literature the introduction of an organoselenium or an organosulfur moiety in a steroid could provide new interesting pharmaceutically active entities exerting anticancer and antimicrobial activities. In this optic, new synthetic strategies to efficiently prepare this class of compounds could be strongly desirable.

Selenosteroids - promising hybrid compounds with pleiotropic biological activity: synthesis and biological aspects.

It is established that steroid based agents are an example of compounds obtained from natural patterns and are of great importance due to their application in the prevention and treatment of diseases. Selenosteroids are hybrids formed by attaching Se-moiety to a steroid molecule. In these types of hybrids, selenium can be present as selenide or as a part of selenosemicarbazones, isoselenocyanates, selenourea, etc. Attaching a Se-moiety to a biologically active steroid might enhance the biological properties of both fragments. Available literature indicates that these kinds of hybrids demonstrate significant anticancer activity, which renders them interesting in terms of medical use. In this review, we present various methods of synthesis and demonstrate that seleno-steroid compounds are promising molecules for further pharmaceutical application.

Steroidal Molecular Rotors with 1,4-Diethynylphenylene Rotators: Experimental and Theoretical Investigations Toward Seeking Efficient Properties.

Properly designed molecular rotors with sizable stators and a fast-moving rotator could provide efficient building blocks for amphidynamic crystals. Herein, we report the synthesis of steroidal compounds 1, 2, and 3 and their deuterated analogues 1D, 2D, and 3D envisioned to work as molecular rotors. The obtained compounds were characterized by attenuated total reflection-infrared, Raman, and circular dichroism (CD) spectroscopy measurements. The interpretation of spectra was supported by theoretical calculations using density functional theory methods. The analysis of the most characteristic bands confirmed different molecular dynamics of the rotors investigated. Angle-dependent polarized Raman spectra showed the crystallinity of some samples. Electronic CD (ECD) spectra of compounds 1-3 and their relevant deuterated analogues 1D-3D are identical. The increase of the band intensity with lowering the temperature shows that the equilibrium is shifted to the thermodynamically most stable conformer. ECD spectra simulated at the TDFFT level of theory for compound 3 were compared with experimental results. It was proved that conformer 3a, with a torsion angle of +50°, exhibits the best agreement with the experimental results. Simulated vibrational CD and IR spectra for conformer 3a and its deuterated analogue 3Da also display good agreement with experimental results. In light of our comprehensive investigations, we evidenced that steroidal compounds 1, 2, and 3 can work as molecular rotors.

Synthesis and application of steroidal 22,16ß-carbolactones: A review.

In the plant kingdom, steroidal lactones occur as glycosides, compounds consisting of a sugar moiety linked to a steroid aglycone. Steroidal lactones consist of five fused rings, with a total of 22 carbon atoms. Numerous methods for the preparation of steroidal lactones take advantage of the fact that steroid spirostanes may be degraded from six- to a five-rings structure. One of the most striking features common to reactions of steroid sapogenins is the C22-lactone formation. In the review, different methods for the preparation of steroidal lactones are presented with consideration of the structure of starting material. In addition, examples of lactones used in the synthesis of biologically active compounds and their analogues are described.

The interferon-induced helicase C domain-containing protein 1 gene variant (rs1990760) as an autoimmune-based pathology susceptibility factor.

PURPOSE: Autoimmune diseases are a group of complex diseases localized in multiple organ systems, with a wide spectrum of symptoms and still unclear causes. The aim of the present study was to analyse a possible association of three autoimmune disabilities - Multiple sclerosis (MS), LADA diabetes and Graves' disease (GD) with single nucleotide polymorphism (SNP; rs1990760) in the IF IH1 gene (also known as a melanoma differentiation-associated protein 5 - MDA5) within the Polish population. An additional goal was also to look for a correlation between this polymorphism and different clinical patient-related factors. MATERIALS AND METHODS: The study population consisted of four groups of 944 unrelated Polish origin Caucasian patients - 324 with GD, 171 with MS, 49 with LADA diabetes and 400 healthy subjects as a control group. The SNP analysis was performed using the allelic discrimination technique. RESULTS & CONCLUSIONS: There were significant associations of risk T allel of the analyzed polymorphism with all studied autoimmune diseases (GDOR = 1.34, p = 7.02e-03; MSOR = 1.36, p = 2.17e-02; LADA - OR = 3.36, p = 8.73e-07). We also found that the frequency of CT and TT genotypes of the rs1990760 IFIH1 gene only in females (with LADA, GD, MS) was significantly higher than those in the female control group (47%, 41% vs 44%, 34%; p = 1.32e-03, p = 4.39e-04; OR = 2.08, 95%CI: (1.33-3.28), OR = 2.29, 95% CI: (1.44-3.65) respectively). Our research has shown significant differences regarding some clinical features (BMI, TRAb, TSH, HbA1C, anti-GAD antibodies) and age at the beginning of the studied autoimmune disabilities. This study showed an association of rs1990760 polymorphism in the IFIH1 gene in the development of GD, LADA diabetes and MS within the Polish population. To our knowledge, this is the first study to investigate the relationship between IFIH1 polymorphisms and the risk of the development of MS and LADA in Poland.

Investigation of lipoic acid - 4-methoxybenzyl alcohol reaction and evaluation of its analytical usefulness.

The presented work is aimed to synthesize a new UV active derivative of α-lipoic acid (ALA) by its esterification with 4-methoxybenzyl alcohol (4-MBA, anise alcohol). The formation of ester was confirmed by 1HNMR, FTIR and UV spectroscopy. The analytical usefulness of the obtained compound for quantification of ALA in food items was examined using HPLC-UV and GC-MS systems. It was found that it is possible to assay ALA in the ester form in the concentration ranges: 5·10-6-1·10-4 mol L-1 by HPLC-UV and 1∙10-7-5∙10-5 mol L-1 by GC-MS techniques. The GC-MS procedure was applied for the determination of ALA in the food samples.

PhSeZnCl in the Synthesis of Steroidal ß-Hydroxy-Phenylselenides Having Antibacterial Activity.

We report here the reaction of in situ prepared PhSeZnCl with steroid derivatives having an epoxide as an electrophilic functionalization. The corresponding ring-opening reaction resulted to be regio- and stereoselective affording to novel phenylselenium-substituted steroids. Assessment of their antibacterial properties against multidrug-resistant bacteria, such as Pseudomonas aeruginosa Xen 5 strain, indicates an interesting bactericidal activity and their ability to prevent bacterial biofilm formation.

Revision of the Structure of N, O-Diacetylsolasodine. Unusual Epimerization at the Spiro Carbon Atom during Acetylation of Solasodine.

The steroidal alkaloid solasodine (1) undergoes inversion of configuration at the C-22 spiro atom when treated with acetic anhydride-pyridine at ambient temperature. The basic solvolysis of the N, O-diacetyl derivative (2) reverses the reaction, yielding the starting solasodine (1). The mechanisms of both processes (acetylation and deacetylation) were studied in terms of possible reaction pathways.

Complex formation equilibria between cholesterol and diosgenin analogues in monolayers determined by the Langmuir method.

The purpose of this study was to investigate the interaction between diosgenin analogues [DioA: diosgenin acetate (DAc) and (25R)-5α,6ß-dihydroxyspirostan-3ß-ol acetate (DSol)] and cholesterol (Ch) monolayers at the air/water interface. The surface tension of pure and mixed lipid monolayers at 22 °C was measured by using the Langmuir method with a Teflon trough and a Nima 9002 tensiometer. The surface tension values were used to calculate the π-A isotherms and to determine the molecular surface areas. The interactions between Ch and each DioA resulted in significant deviations from the additivity rule. The theory described in this work was used to determine the stability constants, the areas occupied by one molecule of Ch-DAc or Ch-DSol, and the complex formation energy (Gibbs free energy) values.

Vibrational spectroscopic characterization of cyclic and acyclic molecular rotors with 1,4-diethynylphenylene-d4 rotators.

The synthesis and characterization of acyclic and cyclic molecular rotors with 1,4-diethynylphenylene-d4 rotators are described. The ATR-FTIR and Raman spectra of acyclic rotor 3 and cyclic rotor 5E were measured and interpreted. A feature of ATR-FTIR spectrum of rotor 5E is a strong two-component band around 1730cm-1 attributed to symmetric and asymmetric stretching vibration of the carbonyl group, while this is not observed in rotor 3. Raman investigation in the wide temperature range of 350 - 10K was carried out. The splitting of Raman bands in the region of stretching vibration of CC double and triple bonds at 170 and 260K for rotor 5E is observed. The splitting of bands is due to changes in molecular structure.

Some observations on solasodine reactivity.

This article presents new transformations of solasodine - a representative steroid alkaloid sapogenin from the Solanum family. Oxidation of N,O-diacetylated solasodine with either NaNO2/BF3·Et2O or t-BuONO/BF3·Et2O resulted in partial degradation of the side chain to (20S)-3ß-acetoxypregn-5-ene-20,16ß-carbolactone (vespertilin acetate). The same starting compound when treated with TMSOTf afforded the corresponding pseudosapogenin after aqueous work-up. However, when the crude reaction mixture was directly subjected to purification on a silica gel column, efficient autoxidation to pregna-5,16-dien-3ß-ol-20-one acetate was observed. One-step synthesis of this important drug intermediate from spirosolan alkaloids may be potentially exploited for large-scale production of steroid hormones.

The synthesis and antifungal activity of (20S)-3ß-acetoxy-5α-pregnane-20,16ß-carbolactone against fluconazole - Resistant Candida cells.

An efficient procedure for the synthesis of (20S)-3ß-acetoxy-5α-pregnane-20,16ß-carbolactone is described. Bactericidal and fungicidal activity of the lactone against different bacteria such as MSSA, MRSA, E. coli ESBL, P. aeruginosa and clinical isolates of Candida spp., in planktonic and biofilm growth stage were assessed. Additionally, the affinity of this new compound to microbial plasma membrane and hemoglobin release from human red blood cells were determined using fluorometric and colorimetric assay, respectively. Our studies revealed that the lactone exhibits strong antifungal activity, and the ability to prevent pathogens' biofilm formation. Additionally, upon lactone treatment a significant affinity to fungal, but not to human cell membranes, indicating suitable biocompatibility was observed.

The Equilibria of Diosgenin-Phosphatidylcholine and Diosgenin-Cholesterol in Monolayers at the Air/Water Interface.

Diosgenin (Dio) has shown many treatment properties, but the most important property is cytotoxic activity in cancer cells. In this study, we investigated monolayers of Dio, cholesterol (Ch), and phosphatidylcholine (PC) at the air/water interface. The measurements were carried with a Langmuir Teflon trough and a Nima 9000 tensiometer program. The surface tension values of pure and mixed monolayers were used to calculate π-A isotherms and determine molecular surface areas. We were able to demonstrate the formation of complexes between Dio and PC and Dio and Ch molecules also. We considered the equilibrium between individual components and the formed complexes. In addition, we established that diosgenin and the lipids formed highly stable 1:1 complexes.

Oxidation of Olefins with Benzeneseleninic Anhydride in the Presence of TMSOTf.

A new oxidizing system for olefins, consisting of benzeneseleninic anhydride and trimethylsilyl triflate, was studied. The highly reactive benzeneseleninyl cation is presumably formed under these conditions. It has been shown that different products are formed with this species depending on the specific structure of olefin. The 1,1-disubstituted olefins afforded mostly α,ß-unsaturated carbonyl compounds. The sterically encumbered tri- or tetrasubstituted olefins yielded 1,2- or 1,4-dihydroxylated products, presumably via four-membered cyclic intermediates.