Cascade Transformation of the Ansamycin Benzoquinone Core into Benzoxazole Influencing Anticancer Activity and Selectivity.

The metal-free cascade transformation of geldanamycin benzoquinone core is proposed at relatively mild conditions. This approach yields new benzoxazole ansamycin antibiotics and enables their functionalization in an atom-economic manner, irrespective of the type of amine used. The analysis of the heterocyclization course reveals the dependence of its rate on the nature of the para-substituent within the benzylamine moiety (EDG/EWG) and the strength of the base. The reduction of the ansamycin core enables an increase in anticancer potency and selectivity.

Modifications of geldanamycin via CuAAC altering affinity to chaperone protein Hsp90 and cytotoxicity.

Functionalization of alkyne (1) and azide (2) derivatives of geldanamycin (GDM) via dipolar cycloaddition CuAAC yielded 35 new congeners (3-37) with C(17)-triazole arms bearing caps of different nature (basic vs. acidic, hydrophilic vs. hydrophobic). Confrontation of biological data (anticancer activity vs. toxicity in normal cells) with lipophilicity (clogP), dissociation constants (Kd) of complexes with Hsp90 and binding modes to Hsp90 revealed SAR in specific subgroups of GDM derivatives. The most potent GDM congeners 14-16, bearing C(17)-triazole-benzyl-halogen arms exhibited the most optimal clogP values of 2.7-3.1 at favourable binding to Hsp90 (KdHsp90 at µM level). The anticancer activity of 14-16 (IC50 = 0.23-0.41 µM) is higher than those of GDM (IC50 = 0.58-0.64 µM) and actinomycin D (ActD, IC50 = 0.62-0.71 µM) in SKBR-3, SKOV-3 and PC-3 cell lines, with a comparable cytotoxicity in healthy cells. The relationship between structure and attractive anticancer potency (IC50 = 0.53-0.74 µM) is also observed for congeners with C(17)-triazole-saccharide or C(17)-triazole-unsaturated arms. In the former, the absolute configuration at C(4) (ᴅ-glucose vs. ᴅ-galactose) whereas in the latter the length of the unsaturated arm influences the cytotoxic effects due to different binding strength (Kd, ΔE) and modes with Hsp90. Among all triazole congeners of GDM that are biologically attractive and exhibit lower toxicity in normal cells than GDM and ActD, the derivative 22, bearing the C(17)-triazole-cinnamyl arm, shows the lowest Kd (Hsp90), optimal clogP = 2.82, the best pro-apoptotic properties in SKBR-3 and SKOV-3 and the best selectivity indices (SI). For the most potent GDM derivatives with C(17)-triazole arm, the docking studies have suggested the importance of the intermolecular stabilization between the arm and the D57 or Y61 of Hsp90.

Outcomes of Open Repair Treatment for Acute Versus Chronic Achilles Tendon Ruptures: Long-Term Retrospective Follow-Up of a Minimum 10 Years-A Pilot Study.

The aim of the present study was to compare repaired Achilles tendon (AT) remodelling, whether its function was restored and what effects the surgery had on our patients' gait cycle in a long-term follow-up study. The study population comprised 30 human subjects treated acutely and chronically for AT ruptures, using the same surgical technique in all cases. The study group was divided into two subgroups regarding the age of their AT injury, i.e., how much time elapsed between the injury and when a correct diagnosis was made and when adequate treatment was applied. Following these criteria, persons presenting at less than 4 weeks postinjury were classified as acute rupture (AR) patients and those presenting at more than 4 weeks after injury were grouped as chronic rupture (CR) patients. Both patient groups were operated on using a surgical method favoured at least a decade ago, i.e., open repair through a posteromedial approach. The AT was augmented with a plantaris longus tendon autograft, followed by suturing using the pull-out suture technique. The results were measured using clinical, ultrasonographic (US) and pedobarographic methods. Our ultrasonographic and pedobarographic findings revealed differences between both patient groups, thus indicating that delayed surgery had negative impacts on treatment success, however, with good long-term functional score outcomes in both patient groups. Nevertheless, delayed treatment of AT ruptures did not leave individual gait phases unaffected, as it also affected the plantar surface and balance performance of the affected limb. As per the results, the Achilles tendon manifested decreased capacity following delayed treatment; however, its long-term functional outcomes were favourable, irrespective of whether it was for acute or chronic patients.

Cascade synthetic strategies opening access to medicinal-relevant aliphatic 3- and 4-membered N-heterocyclic scaffolds.

Cascade reactions are often 'employed' by nature to construct structurally diverse nitrogen-containing heterocycles in a highly stereoselective fashion, i.e., secondary metabolites important for pharmacy. Nitrogen-containing heterocycles of three- and four-membered rings, as standalone and bicyclic compounds, inhibit different enzymes and are pharmacophores of approved drugs or drug candidates considered in many therapies, e.g. anticancer, antibacterial or antiviral. Domino transformations are in most cases in line with modern green chemistry concepts due to atom economy, one-pot procedures often without use the protective groups, time-saving and at markedly lower costs than multistep transformations. The tandem approaches can help to obtain novel N-heterocyclic scaffolds, functionalized according to structural requirements of the target in cells, taking into account the nature of functional group and stereochemistry. On the other hand cascade strategies allow to modify small N-heterocyclic rings in a systematic way, which is beneficial for structure-activity relationship (SAR) analyses. This review is focused on the biological relevance of the N-heterocyclic scaffolds with smaller 3- and 4-membered rings among approved drugs and leading structures of drug candidates. The cascade synthetic strategies offering N-heterocyclic scaffolds, at relatively good yields and high stereoselectivity, are discussed here. The review covers mainly years from 2015 to 2021.

Factors Affecting Dental Caries Experience in 12-Year-Olds, Based on Data from Two Polish Provinces.

(1) Background: Dental caries is a chronic disease that affects a child's dentition from the first stages of life. Several factors contribute to the development of the disease, including an improper diet. This cross-sectional study aimed to identify risk factors of dental caries in 12-year-old adolescents from Greater Poland and Lubusz Provinces (Poland). (2) Material and methods: The research was conducted in adolescents from five primary schools. A questionnaire consisted of close-ended questions on socioeconomic characteristics on family, diet, and oral hygiene habits. An assessment of the dentition was carried out in accordance with World Health Organization (WHO) recommendations. In addition to cavitated carious lesions, incipient caries lesions were noted according to the International Caries Detection and Assessment System, adapted for epidemiological studies (ICDASepiDMFt). (3) Results: The mean number of teeth with untreated caries; removed due to caries; and restored because of caries (DMFt) was 1.52 ± 1.90, while the ICDASepiDMFt index amounted to 2.64 ± 2.55, respectively. Children who did not brush every day had significantly higher odds of having ICDASepiDMFt > 0 than children brushing at least once daily (OR = 10.32, 95% CI = 1.36−78.32, p = 0.0240). Adolescents who drank sweet carbonated drinks every day had significantly higher ICDASepiDMTt than children who drank sweet carbonated drinks less frequently (p = 0.0477). (4) Conclusions: The research revealed that dental caries indices of 12-year-old adolescents from Greater Poland and Lubusz Provinces depend mainly on oral hygiene behaviors. The only significant nutritional factor that differentiated the caries intensity was the daily consumption of sweet carbonated drinks.

Tandem construction of biological relevant aliphatic 5-membered N-heterocycles.

Nature often uses cascade reactions in a highly stereocontrolled manner for assembly structurally diverse nitrogen-containing heterocyclic scaffolds, i.e. secondary metabolites, important for medicinal chemistry and pharmacy. Five-membered nitrogen-containing heterocycles as standalone rings, as well as spiro and polycyclic systems are pharmacophores of drugs approved in various therapies, i.a. antibacterial or antiviral, antifungal, anticancer, antidiabetic, as they target many key enzymes. Furthermore, a large number of pyrrolidine derivatives are currently considered as drug candidates. Cascade transformations, also known as domino or tandem reactions, offer straightforward methods to build N-heterocyclic libraries of the great structural variety desired for drawing SAR conclusions. The tandem transformations are often atom economic and time-saving because they are performed as the one-pot, so no need for purification after each 'virtual' step and the limited necessity of protective groups are characteristic for these processes. Thus, the same results as in classical multistep synthesis can be achieved at markedly lower costs and shorter time, which is in line with modern green chemistry rules. Great advantage of cascade reactions is often reflected in their high regio- and stereoselectivities, enabling the preparing of the heterocyclic compound better fitted to the expected target in cells. This review reveals the biological relevance of N-heterocyclic scaffolds based on saturated 5-membered rings since we showed a number of examples of approved drugs together with the recent biologically attractive leading structures of drug candidates. Next, novel cascade synthetic procedures, taking into account the structure of the reactants and reaction mechanisms, enabling to obtain biological-relevant heterocyclic frameworks with good yields and relatively high stereoselectivity, were reviewed and compared. The review covers the advances of designing biological active N-heterocycles mainly from 2018 to 2021, whereas the synthetic part is focused on the last 7 years.

Structural diversity and biological relevance of benzenoid and atypical ansamycins and their congeners.

Covering: 2011 to 2021The structural division of ansamycins, including those of atypical cores and different lengths of the ansa chains, is presented. Recently discovered benzenoid and atypical ansamycin scaffolds are presented in relation to their natural source and biosynthetic routes realized in bacteria as well as their muta and semisynthetic modifications influencing biological properties. To better understand the structure-activity relationships among benzenoid ansamycins structural aspects together with mechanisms of action regarding different targets in cells, are discussed. The most promising directions for structural optimizations of benzenoid ansamycins, characterized by predominant anticancer properties, were discussed in view of their potential medical and pharmaceutical applications. The bibliography of the review covers mainly years from 2011 to 2021.

Modifications, biological origin and antibacterial activity of naphthalenoid ansamycins.

Covering: 2011 to 2021Structural division of natural naphthalenoid ansamycins, regarding the type of the core and length of the ansa chain, and their biosynthetic pathways in microorganisms are discussed. The great biosynthetic plasticity of natural naphthalenoid ansamycins is reflected in their structural variety due to the alterations within ansa bridge or naphthalenoid core portions. A comparison between the biological potency of natural and semisynthetic naphthalenoid ansamycins was performed and discussed in relation to the molecular targets in cells. The antibacterial potency of naphthalenoid ansamycins seems to be dependent on the ansa chain length and conformational flexibility - the higher flexibility of the ansa chain the better biological outcome is noted.

An Approach to Modify 14-Membered Lactone Macrolide Antibiotic Scaffolds.

A ketolide derivative with (12R)-configuration was obtained via a novel ketene acetal in acidic conditions. The structure of this atypical ß-keto ketene acetal intermediate within the macrocyclic system has been determined by NMR and X-ray methods. The use of basic conditions at an elevated temperature yielded new, doubly α,ß-unsaturated ketone macrolide derivatives with (4E)-configuration as two conformational isomers of folded-in or folded-out conformations.

Regioselective approach to colchiceine tropolone ring functionalization at C(9) and C(10) yielding new anticancer hybrid derivatives containing heterocyclic structural motifs.

The influence of base type, temperature, and solvent on regioselective C(9)/C(10) "click" modifications within the tropolone ring of colchiceine (2) is investigated. New ether derivatives of 2, bearing alkyne, azide, vinyl, or halide aryl groups enable assembly of the alkaloid part with heterocycles or important biomolecules such as saccharides, geldanamycin or AZT into hybrid scaffolds by dipolar cycloaddition (CuAAC) or Heck reaction. Compared to colchicine (1) or colchiceine (2), ether congeners, as e.g. 3e [IC50s(3e) ∼ 0.9 nM], show improved or similar anticancer effects, whereby the bulkiness of the substituents and the substitution pattern of the tropolone proved to be essential. Biological studies reveal that expanding the ether arms by terminal basic heterocycles as quinoline or pyridine, decreases the toxicity in HDF cells at high anticancer potency (IC50s ∼ 1-2 nM). Docking of ether and hybrid derivatives into the colchicine pocket of αGTP/ß tubulin dimers reveals a relationship between the favourable binding mode and the attractive anticancer potency.

Anticancer activity and toxicity of new quaternary ammonium geldanamycin derivative salts and their mixtures with potentiators.

Geldanamycin (GDM) has been modified by different type neutral/acidic/basic substituents (1-7) and by quinuclidine motif (8), transformed into ammonium salts (9-13) at C(17). These compounds have been characterised by spectroscopic and x-ray methods. Derivative 8 shows better potency than GDM in MCF-7, MDA-MB-231, A549 and HeLa (IC50s = 0.09-1.06 µM). Transformation of 8 into salts 9-13 reduces toxicity (by 11-fold) at attractive potency, e.g. MCF-7 cell line (IC50∼2 µM). Our studies show that higher water solubility contributes to lower toxicity of salts than GDM in healthy CCD39Lu and HDF cells. The use of 13 mixtures with potentiators PEI and DOX enhanced anticancer effects from IC50∼2 µM to IC50∼0.5 µM in SKBR-3, SKOV-3, and PC-3 cancer cells, relative to 13. Docking studies showed that complexes between quinuclidine-bearing 8-13 and Hsp90 are stabilised by extra hydrophobic interactions between the C(17)-arms and K58 or Y61 of Hsp90.

Clinical Consequences of Dental Caries, Parents' Perception of Child's Oral Health and Attitudes towards Dental Visits in a Population of 7-Year-Old Children.

This cross-sectional study aimed to assess the occurrence of the consequences of dental caries and factors affecting dental service utilization in a population of 7-year-old children. The research included oral examination of 7-year-old schoolchildren and socio-medical study of their parents/legal caregivers. It was carried out in five primary schools of two provinces, i.e., Greater Poland and Lubusz. Dental examination was performed in accordance with World Health Organization (WHO) recommendations. The socio-medical study consisted of questionnaires with close-ended questions concerning socioeconomic characteristics of the family, reasons and time of the last visit at the dental office, consequences of child's oral health problems, parents' attitude towards dental visits, and parental opinion about their child's teeth. Factors affecting utilization of dental services were statistically analyzed using univariate logistic regression assuming p < 0.05. The pufa index of examined children ranged from 0 to 7 (mean 0.80 ± 1.49), while the dmf index ranged from 0 to 14 (3.86 ± 3.32). Low financial burden of oral health expenditures and university education of at least one of the parents significantly increased the chance of visiting a dentist despite lack of pain (OR = 3.0 and 2.5, respectively). In spite of the availability of free dental care for children, socioeconomic factors still determine the utilization of dental services in Poland. Poor oral health status of examined population and negligence of regular dental check-ups emphasize a need to strengthen oral health literacy of parents and children, promoting proper attitudes towards dental care.

Synthesis, structure and anticancer activity of new geldanamycin amine analogs containing C(17)- or C(20)- flexible and rigid arms as well as closed or open ansa-bridges.

The nucleophilic attack of amines at C(17) or C(17)/C(20) positions of geldanamycin's (GDM) benzoquinone, via initial 1,4-Michael conjugate addition mechanism, yield new analogs with closed or open ansa-bridges (1-31), respectively. X-ray structures of analogs 22 and 24 reveals an unexpected arrangement of the ansa-bridge in solid (conformer B), that is located between those of conformers A, prevailing in solution (trans-lactam), and C, crucial at binding to Hsp90 (cis-lactam). The structure of a new-type conformer B allows to better understand the molecular recognition mechanism between the GDM analogs and the target Hsp90. Combined analysis of: anticancer test results (SKBR-3, SKOV-3, PC-3, U-87, A-549) and those performed in normal cells (HDF), KD values and docking modes at Hsp90 as well as clogP parameters, reveals that the rigid C(17)-arm (piperidyl, cyclohexyl) with a H-bond acceptor as carbonyl group together with a lipophilicity clogP∼3 favor high potency of analogs, even up to IC50 ∼0.08 µM, at improved selectivity (SIHDF > 30), when compared to GDM. The most active 25 show higher anticancer potency than 17-AAG (in SKOV-3 and A-549) as well as reblastatin (in SKBR-3 and SKOV-3). Opening of the ansa-bridge within GDM analogs, at the best case, decreases activity (IC50∼2 µM) and toxicity in HDF cells (SIHDF∼2-3), relative to GDM.

Synthesis and Antibacterial Activity of New N-Alkylammonium and Carbonate-Triazole Derivatives within Desosamine of 14- and 15-Membered Lactone Macrolides.

Desosamines of azithromycin (AZM) and clarithromycin (CLA) were modified by N-alkylation or nucleophilic substitution at the carbonyl/CuAAC sequence. Biological studies revealed a higher antibacterial potency of quaternary N-alkylammonium bromides of CLA as compared to AZM. SAR studies of CLA salts, including biological, conformation and molecular-docking analysis, enriched by physicochemical parameters, showed the importance of less bulky and unsaturated substituent for an efficient docking mode at the ribosomal tunnel and good antibacterial potency against clinical and standard Streptococcus pneumoniae and Streptococcus pyogenes strains (MICs 0.25 or 0.5 µg/mL). These CLA salts also have an at least threefold lower cytotoxicity than reference antibiotics at comparable antibacterial activity against the S. pneumoniae clinical strain. Differences in antibacterial effects noted for AZM and CLA salts bearing less bulky N-substituents can be better understood when their binding modes in the ribosomal tunnel are considered rather than their common low lipophilicity and excellent water solubility.

Stress radiography in the diagnosis and assessment of the outcomes of surgical treatment of chronic anterolateral ankle instability.

BACKGROUND: In about 20-40% of patients, damage to the soft tissues surrounding the ankle leads to recurrent and chronic pain with signs of instability of the talus in the tibiofibular fork. PURPOSE: The aim of the study was to assess the usefulness of stress X-ray images in the diagnosis of long-term outcomes of conservative versus surgical treatment of anterolateral ankle instability. MATERIAL AND METHODS: Thirty patients with chronic ankle instability (CAI) were recruited for the study. The participants were divided into two groups. The first group consisted of 15 non-operatively treated individuals diagnosed with anterior talofibular ligament (ATFL) damage. The second group consisted of 15 patients who had undergone surgical reconstruction of the ATFL. In both groups of patients, the contralateral normal limb was used as a control. In all patients, anteroposterior and lateral view stress radiographs of both ankles were taken using the TELOS Stress Device (GA - III/E, Hungen, Germany). RESULTS: Statistical tests showed that the surgically treated patients had a significantly greater ankle stability compared to the non-surgically treated patients (P = 0.001 for talar tilt angle and P = 0.009 for anterior drawer distance). The results obtained in this study indicate that this method can also be used in postoperative assessment of the function of the reconstructed lateral ankle ligaments. CONCLUSIONS: Stress radiography is a reliable and safe tool for diagnosing CAI. This imaging method is an objective instrument that can be successfully used in postoperative assessment of the function of the reconstructed ATFL.

14- and 15-membered lactone macrolides and their analogues and hybrids: structure, molecular mechanism of action and biological activity.

Classical macrolides containing 14- or 15-membered aglycone lactone rings are basic and widely used antibacterial agents in clinical therapy. Macrolides exert antibacterial effects by blocking the exit peptide tunnel during translation at bacterial ribosomes. However, one macrolide occasionally manifests different docking modes or binding sites at ribosomes, which presents difficulties in designing macrolide modifications. In addition to their medical importance, lactone macrolides possess interesting chemistry driven by numerous intramolecular interactions or cascade transformations. Lactone macrolides are potent antibacterial/bactericidal agents, whereas derivatives of macrolactone antibiotics possess anti-malarial, anti-cancer, anti-inflammatory, and anti-leishmanial activity, and can regulate gastrointestinal motility. Hence, it is important to develop novel semi-synthetic and completely synthetic approaches for functionalising lactone cores of different macrolides and their structural analogues such as ketolides, fluoro-ketolides, acylides, and hybrids containing steroids, nucleosides, heterocycles, and peptides. Here, we discuss modifications that generate novel macrolides with enhanced biological potency and improved molecular mechanisms of action. The bibliography presented in this review contains references to articles and patents that were published and filed from 2009 to mid-2019.

Specific Interactions between Rifamycin Antibiotics and Water Influencing Ability To Overcome Natural Cell Barriers and the Range of Antibacterial Potency.

Rifamycins are a group of macrocyclic antibiotics mainly used for the treatment of various bacterial infections including tuberculosis. Spectroscopic studies of rifamycins evidence the formation of temperature- and solvent-dependent equilibria between A-, B-, and C-type conformers in solutions. The B- and C-type conformers of rifamycin antibiotics are exclusively formed in the presence of water molecules. A- and B-type conformers exhibit a hydrophilic and "open" ansa-bridge nature whereas the C-type conformer is more lipophilic due to the presence of a "closed" ansa-bridge structure. The involvement of the lactam moiety of the ansa-bridge in intramolecular H-bonds within rifapentine and rifampicin implicates the formation of a more hydrophilic A-type conformer. In contrast to rifampicin and rifapentine, for rifabutin and rifaximin, the "free" lactam group enhances conformational flexibility of the ansa-bridge, thereby enabling interconversion between A- and C-type conformers. In turn, an equilibrium between A- and C-type conformers for rifamycins improves their adaptation to the changing nature of bacteria cell membranes, especially those of Gram-negative strains and/or to efflux pump systems.

Antifungal, anticancer, and docking studies of colchiceine complexes with monovalent metal cation salts.

Complexes of colchiceine with monovalent cation perchlorates and iodides have been obtained and characterized by spectroscopic methods. DFT and spectroscopic studies reveal that the dihedral angle ω1-1a-12-12a , crucial for colchicine biological mechanism of action, that is, binding to tubulins depends on the diameter of the complexed metal cation. Biological tests indicated no antifungal properties of colchicine (it was active only toward A.pullulans), in contrast to its derivative-(colchiceine). Complexation of colchiceine with metal cations improved significantly the antifungal potency, even below MIC <1 µg/ml. The colchiceine complexes were more potent than colchiceine, and some of them were even more potent than the fungicidal standard IPBC. The highest potency of colchiceine complexes was noted against A. pullulans (MIC = 0.5 µg/ml). In contrast to the findings concerning antifungal potency, the anticancer studies showed complexes of colchicine more active (~IC50  = 2 nM) than those of colchiceine (~IC50  = 6 µM). MDA-MB-231 breast cancer cell lines and human lung fibroblasts CCD39Lu were also tested.

Synthesis, docking and antibacterial studies of more potent amine and hydrazone rifamycin congeners than rifampicin.

New rifamycin congeners (1-33) with incorporated amine and hydrazone substituents leading to lipophilic and/or basic nature and altered rigidity of modified C(3) arm were synthesized and structurally characterized in detail. NMR spectroscopic studies at different temperatures indicate two types of structures of rifamycin congeners that are realized in solution: zwitterionic and non-ionic forms in dependence of the basicity of modified C(3) arm. The presence of rifamycin congeners in these two possible forms has a significant impact on the physico-chemical parameters such as lipophilicity (clogP) and water solubility and different binding mode of the C(3) arm of antibiotic at RNAP binding pocket (molecular target) leading to different antibacterial potency. The highest antibacterial potency against S. aureus (including MRSA and MLSB strains) and S. epidermidis strains, even higher than reference rifampicin (Rif) and rifaximin (Rifx) antibiotics, was found for rifamycin congeners bearing at the C(3) arm relatively rigid and basic substituents (bipiperidine and guanidine groups). These modifications provide favorable docking mode and excellent water solubility resulting in high potency (MICs 0.0078 µg/mL what gives ∼ 8.5 nM), irrespective whether rifamycin congener is a tertiary amine (15) or hydrazone (29). In turn, for a higher antibacterial potency of rifamycin congeners against E. faecalis strain (MICs 0.5 µg/mL that is 0.6 µM) as compared to Rif and Rifx, the most crucial factors are: bulkiness and the lipophilic character of the end of the C(3) rebuilt arm.

The mechanism of activation of amidobenzylidene ruthenium chelates - latent catalysts of olefin metathesis.

Amidobenzylidene ruthenium chelates - latent catalysts of olefin metathesis can be easily activated by the addition of Brønsted or Lewis acids. Their activation in the presence of hydrogen chloride involves the formation of catalytically active trans-dichloro carbamatobenzylidene ruthenium chelates.