The IL-6 signalling pathway plays a significant role in the progression and development of squamous cell carcinoma (SCC). Bazedoxifene is a potent inhibitor of IL-6R; however, its efficacy is limited by its low solubility in water (0.54 mg/mL). In contrast, BAZE-X1, a formulation of bazedoxifene using sulfobutylether-β-cyclodextrin (SBECD), exhibits enhanced solubility (38 mg/mL). The preparation of this formulation was confirmed through X-ray diffraction and differential scanning calorimetry, while the structure of the bazedoxifene complex with SBECD was elucidated using 1D and 2D NMR spectroscopy. Although BAZE-X1 (10.0 µmol/L; 5.3 µg/mL) demonstrated no toxicity toward SCC cell lines (SCC13, LLSCC1, and FaDu), its effect on IL-6-dependent proliferation was significantly better than that of tocilizumab. The reduction in nuclear accumulation of pSTAT3 (Ser727) by BAZE-X1 was confirmed using In-Cell Western analysis. In the scratch assay, BAZE-X1, like tocilizumab, exhibited antimigratory effects against LLSCC1 but not against SCC13. However, in the context of 3D models (SCC13, LSCLC1, and FaDu), BAZE-X1 (1.0 and especially 5.0 µmol/L; 2.65 and 0.53 µg/mL) displayed a potent antimigration effect.

The combination of cytostatics (e.g., doxorubicin) with natural compounds such as curcumin can be an effective strategy in treating malignancies. Curcumin enhances doxorubicin's anticancer effects, but the solubility of curcumine is very low. Nanofibers are promising drug delivery systems, typically loaded with drugs (into them) during electrospinning. However, modifying their surface with hydrogels containing anticancer agents may increase therapeutic efficacy. Therefore, the modification of nanostrips with curcumin and doxorubicin was a part of this study. The surface of commercially available nanostrips was modified with hydrogels and HP-p-CD containing a doxorubicin and curcumin. Upon incubation of these modified nanostrips in a buffer solution, the majority of the loaded agents were released, achieving concentrations of 190 & micro;M for curcumin and 15 & micro;M for doxorubicin. These concentrations were sometimes higher than the effective doses represented by the IC50 values for colorectal cancer cell lines (DLD1, HCT116, LS147, and SW620) and pancreatic cancer cell lines (PANC1, Patu8902, AsPC1, and BxPC3). In addition, a combined dose of 27 & micro;M curcumin and 1 & micro;M doxorubicin resulted in the disintegration of primary human colorectal carcinoma organoids.

The targeting of epigenetic factors, particularly TET proteins (ten-eleven translocation methylcytosine dioxygenases), has emerged as a significant focus in medicinal and biological research. Recent findings indicate that iron chelators possess substantial potential for inhibiting TET activity. In this study, we synthesized two 2-(hetero)aryl-1H-perimidines (perimidine 1 and 2) with iron(II) binding properties. The results show that these derivatives, particularly 2, exhibit notable inhibitory activity and selectivity for the TET1 protein, with an IC50 value of 1.02 mu M, in contrast to TET2, which has an IC50 value of 13.23 mu M.

IntroductionHead and neck cancers (HNC) remain a significant clinical challenge, particularly due to their association with chronic inflammation triggered by tobacco carcinogens and human papillomavirus (HPV) infection. Persistent activation of proinflammatory and proangiogenic pathways, including nuclear factor kappa B (NF-kB), interleukin 6 (IL-6), and interleukin 8 (IL-8), plays a crucial role in tumor progression.MethodsIn this study, we synthetized ruthenium-enhanced curcumin derivatives (complexes 3 and 4) and study their anti-inflammatory and anticancer properties by using HNC cell lines.ResultsComplex 3 demonstrated potent cytotoxic and antiproliferative effects across both HPV-negative and HPV- positive HNC cell lines, while complex 4 showed selectivity toward oral squamous cell carcinoma (OSCC). Both complexes exhibited cytostatic and migrastatic activities. Importantly, treatment with these complexes significantly suppressed NF-kB activity and reduced IL-6 and IL-8 levels more effectively than native curcumin.DiscussionThese findings highlight their potential not only as stand-alone therapeutic agents but also as adjuvants in combination therapies for HNC.

The preparation, isolation, and identification of dinitro, trinitro, and tetranitro metalloporphyrins bearing nitro groups on pyrrole rings-desirable intermediates that enable fundamental changes in the chemico-physical properties of the porphyrin core-are studied. All six possible dinitro-Ni-TPP isomers are formed; three are isolated as pure substances using column chromatography, while the remaining three are inseparable. All possible trinitro-Ni-TPP isomers are prepared and isolated, except for the 2,7,13 isomer. Attempts to synthesize tetranitro-Ni-TPP either through direct nitration of metalloporphyrin or via direct condensation of the nitropyrrole building blocks are unsuccessful. The molecular structures are unambiguously identified using 2D NMR experiments. Some experimental observations, though not all, are consistent with quantum chemical calculations performed on the geometry, energy, Fukui indices, dipole moments of nitroporphyrins, and the energy of nitration intermediates.

Exosomes, nanosized extracellular vesicles released by various cell types, are intensively studied for the diagnosis and treatment of cancer and neurodegenerative diseases, and they also display high usability in regenerative medicine. Emphasizing their diagnostic potential, exosomes serve as carriers of disease-specific biomarkers, enabling non-invasive early detection and personalized medicine. The cargo loading of exosomes with therapeutic agents presents an innovative strategy for targeted drug delivery, minimizing off-target effects and optimizing therapeutic interventions. In regenerative medicine, exosomes play a crucial role in intercellular communication, facilitating tissue regeneration through the transmission of bioactive molecules. While acknowledging existing challenges in standardization and scalability, ongoing research efforts aim to refine methodologies and address regulatory considerations. In summary, this review underscores the transformative potential of exosomes in reshaping the landscape of medical interventions, with a particular emphasis on cancer, neurodegenerative diseases, and regenerative medicine.

The molecular hybridization of three pharmacophores—benzothiazole, cyclobut-2-ene-1, 2-dione, and hydrazone—has led to the development of novel compounds with diverse biological activities, including anticancer, anti-inflammatory, and antiviral properties. Among these, certain derivatives demonstrated potent and selective anticancer activity, particularly in CCRF-CEM and HCT116 cell lines. Some derivatives also emitted green fluorescence, enabling organelle-specific imaging by fluorescence microscopy. In U2OS cells, these fluorescent derivatives induced cytoplasmic vacuolization within 24 hours, suggesting a non-apoptotic cell death mechanism consistent with oncosis, characterized by energy depletion, increased membrane permeability, organelle swelling, and protein denaturation. Various microscopy techniques were employed to explore the fluorescent features of target organelles in U2OS cells. Compound 6, with its stable green fluorescence, was selected for detailed analysis. Mitochondria were visualized using MitoTracker dye, with colocalization assessed via Pearson correlation. Mitochondrial membrane potential was evaluated using TMRM dye, which accumulates only in cells with intact membrane potential. Endoplasmic reticulum colocalization with cytoplasmic vacuoles was analyzed using mCherry-tagged calreticulin in a recombinant U2OS cell line. Lysosomal integrity and vacuole colocalization were examined with LysoTracker dye. To determine if cytoplasmic vacuolization is depended on caspase activation, cells were treated with compound 6 in the presence or absence of the pancaspase inhibitor Z-VAD-FMK, and caspase-3/PARP-1 cleavage was assessed by Western blot. The FUCCI reporter system was used for cell cycle analysis. Compound 6 strongly colocalized with mitochondria and induced mitochondrial dysfunction, ER vacuolization, and oncosis-like non-apoptotic cell death. This suggests that compound 6 exerts anticancer effects by targeting multiple cellular compartments, involving ER stress and mitochondrial disruption. The oncosis-like pathway is significant as it may bypass apoptosis resistance, a key hurdle in cancer therapy. Additionally, compound 6 reduced LysoTracker staining, indicating lysosomal dysfunction. Cell cycle analysis revealed a concentration-dependent G2/M arrest, further supporting its anticancer efficacy. This work was supported by the Technology agency of the Czech Republic project PERMED: T2BA (TN02000109) and the National Institute for Cancer Research (Programme EXCELES, ID Project No. LX22NPO5102). Infrastructure was supported by projects CZ-OPENSCREEN (LM2023052) and EATRIS-CZ (LM2023053). We also acknowledge support from Charles University projects [SVV260637; SVV260521; UNCE 204064; Cooperation ONCO] and IGA_LF_2024_038. Alzbeta Srovnalova, Robert Kakplanek, Zdenek Kejlik, Jan Hajduch, Pavel Martasek, Josef Srovnal, Sona Gurska, Katerina Jecmenova, Petr Dzubak, Marian Hajduch, Milan Jakubek. Oncosis-inudcing hydrazones complexes with selective anticancer activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 2825.

We report the design, synthesis, and biological evaluation of novel hybrid anticancer agents bearing three pharmacophores in one molecule: benzothiazole, cyclobut-2-ene-1,2-dione and hydrazone moieties. Several derivatives have demonstrated potent anticancer activity and high selectivity towards T-lymphoblastic leukaemia (CCRF-CEM) and colorectal cancer (HCT116) cell lines. The effects of in situ formed metal complexes on anticancer activity were investigated, revealing that the Fe(III) complexes of some derivatives were more active than the parental ligands, whereas the Cu(II) and Zn(II) complexes did not enhance cytotoxicity. However, compound 6 exhibited a decrease in cytotoxic activity upon the addition of iron, suggesting that the balance between extracellular and intracellular metal chelation is crucial for the anticancer activity of these compounds. Fluorescence microscopy using U2OS cells revealed the colocalization of specific derivatives with cellular components, including mitochondria (Pearson's coefficient for colocalization yielded positive correlation values of 0.85 and 0.83 for compound 6 at two concentrations), accompanied by a reduced mitochondrial membrane potential, endoplasmic reticulum vacuolization and reduction in lysosomal integrity. Moreover, cell cycle analysis revealed that compound 6 induced concentration-dependent G2/M cell cycle arrest, further contributing to its anticancer activity. These results imply the potential of these compounds to induce nonapoptotic, oncosis-like cell death. Lipinski's rule of five analysis, along with calculated drug-likeness and drug-score factors, indicates that most of these compounds have properties compatible with oral bioavailability and potential for further development. This study presents a promising new class of hybrid anticancer agents with a unique mechanism of action and favourable drug-like properties.

Immune checkpoints regulate the immune system response. Recent studies suggest that flavonoids, known as phytoestrogens, may inhibit the PD-1/PD-L1 axis. We explored the potential of estrogens and 17 Selective Estrogen Receptor Modulators (SERMs) as inhibiting ligands for immune checkpoint proteins (CTLA-4, PD-L1, PD-1, and CD80). Our docking studies revealed strong binding energy values for quinestrol, quercetin, and bazedoxifene, indicating their potential to inhibit PD-1 and CTLA-4. Quercetin and bazedoxifene, known to modulate EGFR and IL-6R alongside estrogen receptors, can influence the immune checkpoint functionality. We discuss the impact of SERMs on PD-1 and CTLA-4, suggesting that these SERMs could have therapeutic effects through immune checkpoint inhibition. This study highlights the potential of SERMs as inhibitory ligands for immune checkpoint proteins, emphasizing the importance of considering PD-1 and CTLA-4 inhibition when evaluating SERMs as therapeutic agents. Our findings open new avenues for cancer immunotherapy by exploring the interaction between various SERMs and immune checkpoint pathways.

Abstract Mitochondrial dysregulation plays a significant role in the carcinogenesis. On the other hand, its destabilization strongly represses the viability and metastatic potential of cancer cells. Photodynamic and photothermal therapies (PDT and PTT) target mitochondria effectively, providing innovative and non-invasive anticancer therapeutic modalities. Cyanine dyes, with strong mitochondrial selectivity, show significant potential in enhancing PDT and PTT. The potential and limitations of cyanine dyes for mitochondrial PDT and PTT are discussed, along with their applications in combination therapies, theranostic techniques, and optimal delivery systems. Additionally, novel approaches for sonodynamic therapy using photoactive cyanine dyes are presented, highlighting advances in cancer treatment.

Octahydroxy bis-porphyrin Troger's base (TB) and octahydroxy porphyrin-chlorin spiro-Troger's base (spiroTB) derivatives were successfully prepared from their Ni(II) complexes utilizing an optimized demetalation method using H2SO4-TFA followed by demethylation via BBr 3 . The synthesized TB and spiroTB were then in in vitro experiments for their utility in photodynamic therapy (PDT) of cancer. Compared to temoporfin ( m THPC, Foscan (R)), the chlorin derivative used in PDT of cancer, TB and spiroTB were localized in lysosomes instead of the endoplasmic reticulum and were found to possess enhanced biocompatibility. Cell culture studies were performed on TRAMP-C2 (prostate cancer), HeLa (cervical cancer), and MRC-5 (non cancerous) cell lines. Both TB and spiroTB displayed negligible dark toxicity but increased production of reactive oxygen species (ROS) when illuminated. In addition, spiroTB displayed significant phototoxicity in TRAMP-C2 cells (IC50, light 0.7 mu M after 24 h incubation). Despite the phototoxicity of spiroTB being lower than that of m THPC (IC50, light 0.02 mu M), due to the low dark toxicity (IC50, dark > 100 mu M), the therapeutic factor of spiroTB ( >150) is six times higher than that of m THPC (25).

TET proteins (methylcytosine dioxygenases) play an important role in the regulation of gene expression. Dysregulation of their activity is associated with many serious pathogenic states such as oncological diseases. Regulation of their activity by specific inhibitors could represent a promising therapeutic strategy. Therefore, this review describes various types of TET protein inhibitors in terms of their inhibitory mechanism and possible applicability. The potential and possible limitations of this approach are thoroughly discussed in the context of TET protein functionality in living systems. Furthermore, possible therapeutic strategies based on the inhibition of TET proteins are presented and evaluated, especially in the field of oncological diseases.

A simple, sensitive and quick HPLC method was developed for the determination of ketoprofen in cell culture media (EMEM, DMEM, RPMI). Separation was performed using a gradient on the C18 column with a mobile phase of acetonitrile and miliQ water acidified by 0.1 % (v/v) formic acid. The method was validated for parameters including linearity, accuracy, precision, limit of quantitation and limit of detection, as well as robustness. The response was found linear over the range of 3-100 μg/mL as demonstrated by the acquired value of correlation coefficient R2=0.9997. The described method is applicable for determination of various pharmacokinetic aspects of ketoprofen in vitro.

Pentamethinium indolium salts are promising fluorescence probes and anticancer agents with high mitochondrial selectivity. We synthesized two indolium pentamethinium salts: a cyclic form with quinoxaline directly incorporated in the pentamethinium chain (cPMS) and an open form with quinoxaline substitution in the γ-position (oPMS). To better understand their properties, we studied their interaction with mitochondrial phospholipids (cardiolipin and phosphatidylcholine) by spectroscopic methods (UV-Vis, fluorescence, and NMR spectroscopy). Both compounds displayed significant affinity for cardiolipin and phosphatidylcholine, which was associated with a strong change in their UV-Vis spectra. Nevertheless, we surprisingly observed that fluorescence properties of cPMS changed in complex with both cardiolipin and phosphatidylcholine, whereas those of oPMS only changed in complex with cardiolipin. Both salts, especially cPMS, display high usability in mitochondrial imaging and are cytotoxic for cancer cells. The above clearly indicates that conjugates of pentamethinium and quinoxaline group, especially cPMS, represent promising structural motifs for designing mitochondrial-specific agents.

Treatment of metastatic cancer is one of the biggest challenges in anticancer therapy. Curcumin is interesting nature polyphenolic compound with unique biological and medicinal effects, including repression of metastases. High impact studies imply that curcumin can modulate the immune system, independently target various metastatic signalling pathways, and repress migration and invasiveness of cancer cells. This review discusses the potential of curcumin as an antimetastatic agent and describes potential mechanisms of its antimetastatic activity. In addition, possible strategies (curcumin formulation, optimization of the method of administration and modification of its structure motif) to overcome its limitation such as low solubility and bioactivity are also presented. These strategies are discussed in the context of clinical trials and relevant biological studies.

Hydrofluoroethers (HFEs) represent a new family of promising engineering fluids suitable for technical cleaning and cooling of electronic and magnetic devices or as admixtures in refrigerant blends. Here, we report accurate data for the liquid density at 0.1 MPa and temperatures from 273.15 K to 343.15 K for a series of five HFEs, namely, HFE-7000, HFE-7100, HFE-7200, HFE-7300, and HFE-7500. A highly sensitive vibrating tube densimeter with a borosilicate glass U-tube calibrated according to the procedure by Prokopová et al. (J Chem Thermodyn 173:106855, 2022) provided density data with an expanded uncertainty ( k=2 ) of 0.13 kg·m^-3 . Influences such as sample degassing, water content, or sample temperature before its dosing into the densimeter are discussed. Thanks to the high sensitivity of the used densimeter, an unexpected shift in the density of different HFE-7100 and HFE-7200 liquid samples was detected. Unlike other HFEs, HFE-7100 and HFE-7200 are mixtures of two hardly separable isomers, which were so far considered having identical thermophysical properties. Utilizing nuclear magnetic resonance spectroscopy, the ratio of n -isomer and iso -isomer was inspected for various liquid samples. In the range of iso -isomer mole fraction from 0.61 to 0.77, the new measurements revealed density differences of more than 5 kg·m^-3 in case of HFE-7100 and of about 3 kg·m^-3 in case of HFE-7200. Consequently, for some applications, the properties of different HFE isomers cannot be considered identical. The Rackett-type correlation for the saturated liquid density was fitted using the new and the literature data.

Targeting of epigenetic mechanisms, such as the hydroxymethylation of DNA, has been intensively studied, with respect to the treatment of many serious pathologies, including oncological disorders. Recent studies demonstrated that promising therapeutic strategies could potentially be based on the inhibition of the TET1 protein (ten-eleven translocation methylcytosine dioxygenase 1) by specific iron chelators. Therefore, in the present work, we prepared a series of pyrrolopyrrole derivatives with hydrazide (1) or hydrazone (2–6) iron-binding groups. As a result, we determined that the basic pyrrolo[3,2-b]pyrrole derivative 1 was a strong inhibitor of the TET1 protein (IC50 = 1.33 μM), supported by microscale thermophoresis and molecular docking. Pyrrolo[3,2-b]pyrroles 2–6, bearing substituted 2-hydroxybenzylidene moieties, displayed no significant inhibitory activity. In addition, in vitro studies demonstrated that derivative 1 exhibits potent anticancer activity and an exclusive mitochondrial localization, confirmed by Pearson’s correlation coefficient of 0.92.

A series of metalloporphyrin dimers as Tröger's bases 1 or spiro-Tröger's bases 2 was prepared starting from five different C4-symmetry porphyrin derivatives substituted in meso-positions by Ph, 3-MeO-Ph, 4-MeO-Ph, 3,4-(MeO)2-Ph, or 3,5-(MeO)2-Ph. Free-base porphyrins were converted to metalloporphyrins, which were subsequently nitrated with nickel(II), copper(II), or zinc(II) nitrate to give β-nitrometalloporphyrins. These were further reduced to β-aminometalloporphyrins and treated with a methanal equivalent under acidic conditions to selectively obtain Tröger's base 1, spiro-Tröger's base 2, or a mixture of both, in yields up to 41% of 1 and 45% of 2 depending on the reaction conditions used. The ratio of 1 to 2 was influenced by the methanal equivalent used, the strength of the acid, and, above all, the solvent. The presence of a metal ion within the porphyrin core and the use of a chlorinated solvent were found to be essential for the formation of spiro-Tröger's base 2. The molecular structure of spiroTB 2a-Ni2 was proven by electron diffraction.

The total solute retention by a chemically modified stationary phase (CMSP) has been shown several times to be a potential tool for studying the binding abilities of the bound compound. In this article, we present a methodology for the deconvolution of the total retention into structure-specific contributions. Three complementary silica-based CMSPs were prepared: 1) non-modified silica, 2) silica modified by syn-bis-Tröger's base (a molecular tweezer) and 3) silica modified by anti-bis-Tröger's base (a non-tweezer molecule). These were characterized by elemental analysis and Raman spectroscopy, and used to assemble liquid chromatography (LC) columns. The total retention factors were estimated for electron-deficient nitro- and cyano-derivatives of benzene in both normal and reverse elution modes. The total retention factor was considered to be the sum of structure-specific retention factors, each related to the affinity (the binding constant) of a specific structure (the binding site), and its content in the modified silica, as defined for weak-affinity chromatography (WAC). The obtained structure-specific contributions are in line with the binding studies of ligands in solution. They reveal details of the retention mechanism, suggesting a more suitable attachment of ligands, and expose the shortcomings of evaluations based solely on the total retentions.

Multi-orthogonal molecular scaffolds can be applied as core structures of bioactive compounds. Here, we prepared four tri-orthogonal scaffolds based on adamantane or proline skeletons. The scaffolds were used for the solid-phase synthesis of model insulin mimetics bearing two different peptides on the scaffolds. We found that adamantane-derived compounds bind to the insulin receptor more effectively (Kd value of 0.5 μM) than proline-derived compounds (Kd values of 15-38 μM) bearing the same peptides. Molecular dynamics simulations suggest that spacers between peptides and central scaffolds can provide greater flexibility that can contribute to increased binding affinity. Molecular modeling showed possible binding modes of mimetics to the insulin receptor. Our data show that the structure of the central scaffold and flexibility of attached peptides in this type of compound are important and that different scaffolds should be considered when designing peptide hormone mimetics.