Bioactive Research Articles

LL37 complexed to double-stranded RNA induces RIG-I-like receptor signalling and Gasdermin E activation facilitating IL-36γ release from keratinocytes

The Interleukin-36 (IL-36) cytokine family have emerged as important players in mounting an inflammatory response at epithelial barriers and tailoring appropriate adaptive immune responses. As members of the Interleukin-1 superfamily, IL-36 cytokines lack a signal peptide for conventional secretion and require extracellular proteolysis to generate bioactive cytokines. Although the IL-36 family plays an important role in the pathogenesis of plaque and pustular psoriasis, little is known about the release mechanisms of these cytokines from keratinocytes and the physiological stimuli involved. Nucleic acid released from damaged or dying keratinocytes initiates early inflammatory signals that result in the breaking of tolerance associated with psoriasis pathogenesis onset. Cathelicidin peptide, LL37 binds to DNA or double-stranded RNA (dsRNA) and activates a type I Interferon responses in plasmacytoid dendritic cells and keratinocytes. Here, we demonstrate that LL37 binds to dsRNA and induces IL-36γ release from human primary keratinocytes. LL37/dsRNA complexes activate RIG-I-like Receptor signalling, resulting in Caspase-3 and Gasdermin E (GSDME) cleavage. Subsequent GSDME pore formation facilitates IL-36γ release. This response is magnified by priming with psoriasis-associated cytokines, IL-17A and IFNγ. IL-36γ release in this manner is largely independent of cell death in primary keratinocytes and lacked extracellular proteolysis of IL-36γ. Conversely, transfection of keratinocytes directly with dsRNA synthetic analogue, Poly(I:C) induces NLRP1 inflammasome activation, which facilitates IL-36γ expression and release in a GSDMD-dependent manner. Inflammasome-associated cell death also enables extracellular processing of IL-36γ by the release of keratinocyte-derived proteases. These data highlight the distinct responses triggered by dsRNA sensors in keratinocytes. Depending on the inflammatory context and magnitude of the exogenous threat, keratinocytes will release IL-36γ coupled with cell death and extracellular cleavage or release the inactive pro-form, which requires subsequent processing by neutrophil proteases to unleash full biological activity, as occurring in psoriatic skin.Cytoplasmic sensing of dsRNA in keratinocytes mediates IL-36γ release via caspase activity and GSDM pore formation Keratinocytes release IL-36γ upon stimulation with intracellular dsRNA alone or complexed to the psoriasis-associated cathelicidin anti-microbial peptide LL37. Left: Transfected dsRNA triggers NLRP1 inflammasome assembly and IL-1β release, which can enhance IL-36γ expression, resulting in IL-36γ release and extracellular cleavage by released proteases. Right: LL37/dsRNA complexes activate a MDA5-MAVS pathway facilitating the release of IL-36γ through Caspase-3 activation and GSDME pore formation.

Synthesis of Differentially Halogenated Lissoclimide Analogues To Probe Ribosome E-Site Binding.

Halogenated natural products from marine sources often demonstrate potent activity against microorganisms and cancer cell lines. During the last three decades, the lissoclimide class of chlorinated labdane diterpenoids has been characterized with respect to structure and cytotoxic activity. Recently, our laboratories have developed different strategies to produce a broad range of naturally occurring lissoclimides and designed synthetic analogues. This work led to the discovery of a novel halogen-π dispersion interaction between the C2 chloride of chlorolissoclimide and guanine residues in the tRNA exit (E) site of the ribosome. In this study, we aimed to synthesize lissoclimide analogues bearing different substituents in place of the chloride to investigate the importance of the halogen identity for binding, translation inhibition, and cytotoxicity. With previous access to the protio and chloro compounds (haterumaimide Q and chlorolissoclimide), we synthesized two more halogenated variants, fluorolissoclimide and bromolissoclimide, as well as a methylated analogue, methyllissoclimide, to complete a panel of chemical probes for functional and structural studies. Using an integrative approach, we explored the effects of these analogues on the eukaryotic translational machinery in vivo and in vitro. X-ray cocrystal structures with the eukaryotic ribosome were solved for each probe molecule, and the effects on ribosomal thermal stability and FRET-derived ribosome binding constants were determined. Together, these data provide a detailed understanding of the different modes of binding of lissoclimides and insight into their relative activities, which vary according to the substitutions that interact with the eukaryote-specific ribosomal protein eL42. Ultimately, we learned that the presence of a lissoclimide C2-halogen atom─offering a potentially stabilizing halogen-π interaction─appears to facilitate or to synergize with a hydrogen-bonding interaction between the C7-hydroxyl group and the backbone of the ribosomal protein eL42, leading to stronger translation inhibition. We therefore conclude that the C2-halogen and C7-hydroxyl groups are critical contributors to potency, and this idea is borne out in the observations of reduced biological activities in the absence of either group.

Fluorine in the Pharmaceutical Industry: FDA-Approved Fluorine-Containing Drugs in 2024.

Fluorine has become an essential element in the development of modern pharmaceuticals, due to its unique chemical properties which can significantly enhance the biological activity, metabolic stability, and lipophilicity of drug molecules. This review explores recent advancements in the synthesis and application of fluorine-containing drugs approved by the US Food and Drug Administration (FDA) in 2024. These novel drugs demonstrate improved efficacy and safety profiles, addressing a range of therapeutic areas including oncology, infectious diseases, metabolic disorders and genetic disorders that affect the adrenal glands. The incorporation of fluorine atoms into drug candidates has facilitated the development of molecules with optimized pharmacokinetic and pharmacodynamic properties, leading to better patient outcomes. The review further discusses the synthetic methodologies employed, the structural characteristics of these drugs, and their clinical implications, providing insights into the ongoing innovation within the pharmaceutical industry driven by fluorine chemistry.

Cutaneous wound healing functions of novel milk-derived antimicrobial peptides, hLFT-68 and hLFT-309 from human lactotransferrin, and bLGB-111 from bovine β-lactoglobulin

The absence of multi-functional antimicrobial agents in clinical settings hinders cutaneous wound healing. Milk-derived antimicrobial peptides (MAPs) may be the imperative solution to wound repair, combining the dermatic curative properties of antimicrobial peptides with the biological activity of milk. Three novel MAPs, which were hLFT-68 (IAENRADAV) and hLFT-309 (GSPSGQKDLLF) identified in human milk and bLGB-111 (LDTDYKKY) identified in bovine milk in our previous work, were initially investigated for their function in wound healing. In vitro, the antibacterial activity and cellular mechanism of the MAPs were examined. It was found that they presented inhibition for Staphylococcus aureus and Escherichia coli, decreased the secretion of inflammatory factors (IL-1β, IL-6, and TNF-α), and promoted fibroblast and keratinocyte proliferation. An infected wound model was established to evaluate the in vivo anti-inflammatory and regeneration properties of the MAPs. The wound area shrank more rapidly, and the wound inflammation was reduced by MAP treatment. Especially on days 3–5 after mouse modeling, the wound repair rate increased by up to 35%. Furthermore, it was suggested that they encouraged collagen synthesis and deposition, and tissue regeneration. The presented results indicated that MAPs accelerated the recovery of infected wounds, possessing the potential for developing wound-healing therapy.

The antioxidant activity of shaddock peel polysaccharides

The shaddock peel polysaccharide (SPP) has various biological activities. This explored its antioxidant activity. In vitro antioxidant tests, the two SPP components showed good DPPH, hydroxyl radical scavenging ability and anti-lipid peroxidation ability. At a concentration of 3.2 mg/mL, the anti-lipid peroxidation abilities of the two polysaccharides were 21.65% and 18.96%, respectively. SPP-3 had a slightly stronger ability to scavenge hydroxyl radicals than SPP-2. When the polysaccharide concentration reached 3.2 mg/mL, the order of the two polysaccharides’ DPPH scavenging rates was SPP-2 > SPP-3. Therefore, SPP showed strong antioxidant activity and could be used in antioxidant-related applications in the future.

Applications of Bridgehead Heterocycles in Drug Design and Medicinal Chemistry.

Bridged heterocycles are highly relevant in medicinal chemistry and drug discovery due to the unique features associated with their three-dimensional configuration that ensures great scaffold complexity. In general, inserting bridged systems into a chemical structure positively influences the pharmacokinetic (PK) profile of leads, reducing lipophilicity and enhancing metabolic stability. Several optimization studies show that bridged systems often promoted a significant improvement of the small molecule-enzyme binding interaction due to conformational changes within the biological target active site. To date, many drugs including bridged cores are available in the market to cure several diseases. Given the broad range of biological activities of naturally occurring and (semi)-synthetic bridgehead heterocycles, here, we have thoroughly reviewed the rational design and the structure-activity relationship (SAR) studies of the most remarkable bridged compounds developed during the past decade, to highlight both the chemical and biological roles of these motifs.

Spatially resolved genome-wide joint profiling of epigenome and transcriptome with spatial-ATAC-RNA-seq and spatial-CUT&Tag-RNA-seq.

The epigenome of a cell is tightly correlated with gene transcription, which controls cell identity and diverse biological activities. Recent advances in spatial technologies have improved our understanding of tissue heterogeneity by analyzing transcriptomics or epigenomics with spatial information preserved, but have been mainly restricted to one molecular layer at a time. Here we present procedures for two spatially resolved sequencing methods, spatial-ATAC-RNA-seq and spatial-CUT&Tag-RNA-seq, that co-profile transcriptome and epigenome genome wide. In both methods, transcriptomic readouts are generated through tissue fixation, permeabilization and in situ reverse transcription. In spatial-ATAC-RNA-seq, Tn5 transposase is used to probe accessible chromatin, and in spatial-CUT&Tag-RNA-seq, the tissue is incubated with primary antibodies that target histone modifications, followed by Protein A-fused Tn5-induced tagmentation. Both methods leverage a microfluidic device that delivers two sets of oligonucleotide barcodes to generate a two-dimensional mosaic of tissue pixels at near single-cell resolution. A spatial-ATAC-RNA-seq or spatial-CUT&Tag-RNA-seq library can be generated in 3-5 d, allowing researchers to simultaneously investigate the transcriptomic landscape and epigenomic landscape of an intact tissue section. This protocol is an extension of our previous spatially resolved epigenome sequencing protocol and provides opportunities in multimodal profiling.

Discovery of a New Class of Thiazolidin-4-one-Based Inhibitors of Human Dihydroorotate Dehydrogenase: Biological Activity Evaluation, Molecular Docking, and Molecular Dynamics

Discovery of a New Class of Thiazolidin-4-one-Based Inhibitors of Human Dihydroorotate Dehydrogenase: Biological Activity Evaluation, Molecular Docking, and Molecular Dynamics

Symmetry impact of 2,5-bis(cycloalkylsulfenyl)[1,3,4]thiadiazoles on their physicochemical properties, thermal behaviors, and bacterial activity

Regarding the importance of 1,3,4-thiadiazoles (TDs) due to versatile biological activity and for investigation of the symmetry impact on thermal behavior and antibacterial activity of 2,5-bis(cycloalkylsulfenyl)[1,3,4]thiadiazole (R2TD), two new derivatives of symmetric and asymmetric R2TD viz. 2,5-bis(cyclopentylsulfenyl)[1,3,4]thiadiazole (C5TD) and 2-cyclohexylsulfenyl-5-cyclopentylsulfenyl[1,3,4]thiadiazole (C6C5TD), were synthesized through the nucleophilic substitution reaction in greener conditions. The chemical structure of products was elucidated by the spectroscopic analysis. Interestingly, suitable crystals of C5TD could be isolated for single-crystal X-ray diffraction analysis from n-hexane after tedious work, whereas, the asymmetric derivative was a low viscous liquid. Then, the influence of symmetry on the thermal behavior of two new TD derivatives and C6TD was studied by TG-DTG and DSC. The thermal behavior of C5TD in a crystalline state is analyzed in detail for the first time. The pharmacokinetics and toxicology of three derivatives of R2TD were evaluated by two prediction tools of SwissADME and T.E.S.T, respectively. Moreover, the in vitro antibacterial activity of the new TD derivative was evaluated against S. aureus and E. coli as gram-positive and gram-negative bacterial species and compared with the activities of chloromycetin and gentamicin as standard antibiotics by the broth microdilution method.

Microplastic accumulation in respiratory and digestive systems of selected fish from Banten Bay, Indonesia.

Microplastics pose a significant environmental threat as they can be consumed directly or indirectly by various marine organisms. This investigation explores the ecological hazards of microplastic pollution in marine life, with an emphasis on fish in Banten Bay, Indonesia. The sampling spots were in Banten Bay where traditional fishing gears known as "bubu" were used and nets. This study represents an inaugural examination of microplastic contamination in selected fish species including Gambusia affinis, Stolephorus indicus, Scatophagus argus, Epinephelus coioides, Rastrelliger sp., Parapenaeopsis sculptilis, and Leiognathus sp. As a result, microplastics were detected in each fish, with a notable increase in concentrations within the gills (7.85 ± 3.54 items/ind) as opposed to the digestive tract (4.95 ± 2.15 items/ind). Their presence indicated a significant difference (p < 0.01), primarily observed as fragments and filaments. Nine distinct polymer types were identified with FT-IR in fish samples collected from Banten Bay, including polyethylene terephthalate, polyvinyl chloride, polyester, polyurethane, ethylene propylene, polypropylene, polyethylene, polystyrene and polyphenylene sulfide. Microplastics were primarily detected within the size range of 20 to 4510µm. Microplastic contamination in fish causes detrimental implications on aquatic ecosystems and human health, since the particles exist in the food chain, altering the biological activities of fish, and potentially posing hazards to consumers. Considering the closeness of Banten Bay to Indonesia's capital and economic hub, immediate preventive actions are essential to safeguard human health and the ecosystem. Additional investigation into plastic degradation and waste management is crucial for comprehending the origins of contamination. The results establish a foundation for continuous monitoring of microplastic risks in Banten Bay and other swiftly evolving coastal ecosystems in Northeast Asia.

Effect of hydroxy groups on X-ray-induced reactions of azo benzene derivatives.

Caged compounds whose chemical bonds are cleavable by specific stimuli are useful tools for life science research because they facilitate control of various biological activities spatiotemporally. Although caged compounds activatable by hard X-rays can be employed for control in deep tissue owing to the high bio-permeability of X-rays, chemical bond cleavage by ionizing radiation has not been investigated adequately. Previously, we demonstrated that an azo bond tethered to a rhodamine scaffold can be efficiently cleaved by hydrated electrons, which is one of the radiolysis products of water, to release rhodamine. In this study, we synthesized novel azo benzene derivatives, AZO1-4, which can release 3-aminobenzamide (3-ABA), a poly (ADP-ribose) polymerase (PARP) inhibitor, and hydroxy groups or amino groups were introduced into them in order to assess the substituent effect on azo bond cleavage. While the amount of 3-ABA was nearly the same for all the azo compounds, decomposition of azo compounds increased according to the number of hydroxy groups. Furthermore, a methoxyl-radical-adding product was detected from AZO2. These results suggested that the hydroxy group accelerates not azo bond cleavage but the other decomposition pathway.

Anticancer potential of lactoferrin: effects, drug synergy and molecular interactions.

Cancer treatment is among today's most active and challenging research fields. In recent years, significant progress has been made in developing new cancer therapies, including nutraceuticals and natural compounds with anticancer properties. Lactoferrin, a glycoprotein present in mammals, is of significant interest due to its pleiotropic behavior, demonstrating a broad spectrum of biological activities such as antimicrobial, antioxidant, anti-inflammatory, immunomodulatory, and anticancer effects. In this review, we examine the current knowledge of Lf's role in cancer. In addition, it exhibits a synergistic effect along with conventional drugs, potentially enhancing their efficacy and, at the same time, reducing the side effects associated with most traditional therapies. However, it is essential to consider the precise molecular mechanism by which Lf exerts its antitumor activity. Searching interactions with several molecules can provide insight into this mechanism. Additionally, finding lactoferrin receptors can improve the strategies for the specific release of the conjugates. For all these reasons, Lactoferrin becomes a potential therapeutic agent that should be examined in depth.

Engineered anti-HER2 drug delivery nanosystems for the treatment of breast cancer.

Breast cancer stands as the primary cancer affecting women and the second most prevalent cause of cancer-related fatalities in developed nations. Consequently, there is a pressing demand for the advancement of therapeutic strategies that can be seamlessly integrated into clinical applications. We investigated the effectiveness of an encapsulation and decoration strategy employing biodegradable and biocompatible carriers together with 3D collagen-based culture models. Envisioning the use of nano delivery systems for localized regional release, we explored the feasibility of a light-controlled drug release, assisted by optical fibers. PLGA nanoparticles loaded or decorated with trastuzumab (TZ) were synthesized via a double emulsion protocol and characterized by dynamic light scattering, surface plasmon resonance, transmission electron microscopy, atomic force microscopy, and Fourier transform infrared spectroscopy. In vitro biological evaluation was then performed on HER2-positive breast cancer cell line BT-474, examining the effect of nanoformulations on cell viability in 2D and 3D collagen scaffold culture models. Cell cycle, apoptosis, cell morphology and distribution and protein expression were analyzed. Finally, a core-offset optical fiber was fabricated and particles release was studied in vitro using light in batch and microfluidic tests. The nanoparticles displayed uniform and spherical shape, maintaining stability in DMEM for up to seven days. The successful immobilization of TZ was verified. In vitro trials with BT-474 cells in 2D and 3D models revealed that poly(lactic-co-glycolic acid) (PLGA) nanoparticles encapsulated with TZ demonstrated similar or superior biological activity compared to free TZ. Notably, PLGA functionalized with TZ both internally and on the surface exhibited the highest effectiveness in terms of cell viability, increase of apoptosis markers, and inducing cell quiescence. This affirms the pivotal role of PLGA nanoparticles in preserving the integrity of TZ and enhancing its targeted delivery. Furthermore, we propose a breakthrough fiber-optic technology for the less invasive local delivery of PLGA-based nanocarriers that can be effectively used in clinical practice. In conclusion our studies lay the foundation for future advancements in alternative therapeutic tools for localized breast cancer treatment. The integration of advanced carriers, optical fibers, and microfluidics opens up new possibilities for innovative and targeted therapeutic approaches.

Molecular Docking Studies and Biological Activities of Chalcones Targeting Acetylcholinesterase, and Carbonic Anhydrase Isoenzymes

Molecular Docking Studies and Biological Activities of Chalcones Targeting Acetylcholinesterase, and Carbonic Anhydrase Isoenzymes

Exosomes Derived from Metformin-Pretreated BMSCs Accelerate Diabetic Wound Repair by Promoting Angiogenesis Via the LINC-PINT/miR-139-3p/FOXC2 Axis.

Chronic trauma is a prevalent and significant complication of diabetes. Mesenchymal stem cell(MSC)-derived exosomes (Exos) have been reported to accelerate the healing of chronic diabetic wounds. MSCs pretreated with chemical or biological factors were reported to enhance the biological activity of MSC-derived exosomes. Hence, this study investigated the role of exosomes derived from bone marrow mesenchymal stem cells (BMSCs) pretreated with metformin (MET) on diabetic wound healing. The results showed that MET-Exos promoted endothelial cell migration, tube formation, and angiogenesis, leading to accelerated wound healing in diabetic mice. Mechanistically, MET-Exos upregulated LINC-PINT, which, through competitive binding to miR-139-3p, activated FOXC2, a key regulator of angiogenesis. These data reveal that MET-Exos might promote revascularization and wound healing through the LINC-PINT/miR-139-3p/FOXC2 axis, showing its potential as a therapeutic modality for diabetic wounds.

Exploring the potential of Fucoidan from Laminaria japonica: A comprehensive review of its biological activities and benefits for human.

Exploring the potential of Fucoidan from Laminaria japonica: A comprehensive review of its biological activities and benefits for human.

Traditional Herbal Medicine Pithecellobium clypearia (Jack) Benth: Research progress in chemical constituents and pharmacological activities.

Traditional Herbal Medicine Pithecellobium clypearia (Jack) Benth: Research progress in chemical constituents and pharmacological activities.

Hydroxylation and Biological Activity Evaluation of Isosteviol C-12 by P450 BM3.

Isosteviol, a naturally occurring tetracyclic triterpenoid, is a versatile compound with various biological activities. Activation of the inert C-H bonds of isosteviol can increase its potential biological activity, but this C-H activation remains challenging in traditional organic synthesis. Cytochrome P450 BM3 is typically used as a tool enzyme to accomplish this goal. In this study, we used compound similarity analysis, molecular docking and site-specific mutation to successfully achieve P450 BM3 catalytic hydroxylation of isosteviol to 12β-OH isosteviol. Compared to isosteviol, 12B-Oh-isosteviol has enhanced anti-cancer and anti-inflammatory properties. Site-Directed Mutagenesis was performed on eight potential key amino acid mutation sites of the BM3 variant, and the best mutant M05 (F87A/A330W/A82G) in the constructed small library increased the conversion rate of the isosteviol from 59% to 66%.

Formation of self-assembled polyelectrolyte complex derived from BSA and nanogels: a study to optimize processing parameters and preserve protein integrity

Objective The aim of this work was to identify, optimize, and use nondestructive process to develop nano-formulation using polyelectrolyte complexation (PEC) between polymeric nanocarrier and bovine serum albumin. Significance Proteins are mostly degraded during preparation and loading into nano-carriers which hinders success in protein delivery. Method Herein, novel PEC consisting of model protein BSA and nanogels (NGs), were prepared to form self-assembled polyelectrolyte nanocomplexes (BSA/NGs-PEC). The BSA/NGs-PEC were obtained by mixing BSA and nanogels at various weight ratios (1:2, 1:4, 1:5, 1:6, 1:8, 1:10), pH values of solution (1.2, 4.0, 6.0), incubation time (2, 4, 6, 8 h), and stirring rate (without, 100, 200 rpm). The prepared PEC were evaluated for particle size (PS), polydispersity index (PDI), zeta potential (ZP), and percentage of complexation efficiency (%CE). To study insights into structural integrity and biological activity, the SDS-PAGE and esterase activity assay was performed on BSA released from final optimized formulation. Results The optimized parameters were BSA/nanogels mixing ratios at 1:8, pH of complex-forming medium at 4.0, incubation time of 6 h, and stirring rate at 100 rpm. The SDS-PAGE and esterase activity assay revealed that the primary structure and bioactivity, respectively, of BSA was still intact. Conclusion The results suggest that current scheme for optimization has considerable potential for creating protein-based delivery system by using PEC via electrostatic interaction.

Unraveling the molecular mechanism underlying the anticancer activity of CISD2/NAF-144-67.

Unraveling the molecular mechanism underlying the anticancer activity of CISD2/NAF-144-67.