Abstract: Conventional anti-inflammatory therapies, including nonsteroidal anti-inflammatory drugs (NSAIDs) and corticosteroids, remain the mainstay of treatment for inflammatory disorders. However, their long-term use is often limited by adverse effects, insufficient target specificity, and an inability to achieve sustained resolution of chronic inflammation. Advances in the understanding of inflammatory signalling pathways have led to the identification of several promising protein targets for the treatment of chronic inflammatory and autoimmune diseases. This review provides a comprehensive overview of emerging therapeutic protein targets whose dysregulation contributes to inflammation and neuroinflammation, highlighting their molecular mechanisms, therapeutic potential, and current clinical relevance. Key targets discussed include inflammasomes, the JAK–STAT pathway, the IL-17/IL-23 axis, specialized pro-resolving mediators (SPMs), the gut microbiota, epigenetic regulators, microRNAs, microglial activation, astrocyte dysfunction, the sphingosine-1- phosphate (S1P) signalling pathway, gasdermins, transient receptor potential (TRP) channels, and the complement system. Current therapeutic strategies targeting these pathways include smallmolecule inhibitors, monoclonal antibodies, and other emerging biologics designed to achieve selective immunomodulation. Collectively, these advances offer new opportunities for precision medicine by enabling more targeted control of inflammation while preserving essential host immune functions. Furthermore, the integration of multi-target therapeutic approaches, innovative drug delivery systems, and personalized treatment strategies is reshaping the future of anti-inflammatory therapy. Continued efforts to restore immune homeostasis without compromising protective immune responses will be critical for the development of safer and more effective treatments for inflammatory diseases.
Objective: Oysters are abundant in protein and represent a promising raw material for producing bioactive peptides. This work aimed to isolate antithrombotic peptide-rich fractions from Crassostrea hongkongensis meat and evaluate their anticoagulant and antithrombotic effects via in vitro clotting assays and mouse in vivo experiments. Methods: Enzymatic hydrolysis was performed to prepare EC. Sequential ultrafiltration and dialysis were adopted to obtain graded fractions FEC1-FEC5. In vitro clotting time assays were conducted to screen active fractions. FEC5 was further purified by Sephadex G-15 gel filtration to yield four subfractions (F1-F4) with different molecular weight distributions. Chemical composition, molecular spectral features, and molecular weight distribution of EC, FEC5, and F2 were characterized using UV scanning, FTIR spectroscopy, and UHPLC-QTOF-MS. A carrageenan-induced mouse tail thrombosis model was established to evaluate the in vivo antithrombotic potency of FEC5, indicated by IR and PCTE. Results: Among all subfractions, F2 possessed the highest peptide content (90.17%) and prolonged in vitro clotting time to 401.92 s, with anticoagulant activity approaching that of HSPCG (p > 0.05). The high-dose FEC5 group exhibited in vivo antithrombotic activity, with an IR of 41.98% and a PCTE of 43.63%. These indicators were numerically higher than those of the APDG without statistical significance (p > 0.05). UHPLC-QTOF-MS analysis confirmed eight shared peptide components between FEC5 and F2 with defined molecular weights. Characteristic marker ions of FEC5 remained detectable after gel filtration, yet residual system impurities existed in F2, indicating that single-round Sephadex G-15 purification cannot completely remove small-molecule contaminants. Chromatography also altered component distribution and molecular aggregation states. Typical peptide absorption signals and phenylalanine characteristic UV absorption were observed in EC, FEC5, and F2, and FTIR spectra displayed characteristic amide A and amide I bands. Discussion: The peptide-rich fractions from Crassostrea hongkongensis, especially FEC5, show obvious anticoagulant and antithrombotic effects in vitro and in vivo. FEC5 exhibited dosedependent activity, with its efficacy numerically close to aspirin, and high concentrations of FEC5 achieved anticoagulant activity comparable to heparin sodium in vitro. Further exploration is required to reveal its underlying action mechanisms. Conclusion: FEC5 exerted dose-dependent anticoagulant and antithrombotic effects in vitro and in vivo. Further purification effectively improved the activity of its subfraction F2. The peptide-rich fractions from Crassostrea hongkongensis have potential for developing marine antithrombotic functional food ingredients.
Introduction: Protein aggregation is an issue that affects the stability and effectiveness of biopharmaceutical products. Proteins easily undergoes denaturation, due to mechanical agitation and prolonged storage times. Although polysorbates have been extensively used to stabilize proteins, there is still room to identify other highly effective additives. This research aimed to compare the efficacy of hydroxypropyl-β-cyclodextrin (HP-β-CD) in preventing protein aggregation in recombinant human insulin with polysorbate 20 (PS-20), polysorbate 80 (PS-80), and Poloxamer 188 (P188). Methods: Recombinant human insulin formulations with the addition of PS-20, PS-80, P188, or HP-β-CD, or the absence of any additive (control group), were prepared. The prepared samples were subjected to mechanical stress through shaking at 100 rpm for 24 hours. Thereafter, a sixmonth accelerated stability test was performed at 25±2°C/65±5% RH. Protein aggregation and particle formation were measured using size-exclusion high-performance liquid chromatography (SEHPLC) to detect HMWP, dynamic light scattering (DLS) for submicron aggregate size, and light obscuration (LO) of subvisible particles. Results: Mechanical agitation caused protein aggregation in all formulations; however, the degree of aggregation depended on the stabilizer used. HP-β-CD showed the highest stabilization activity, with only a 1.2% increase in HMWP level, whereas the non-stabilized control increased by 12.3%. LO analysis showed that formulations containing HP-β-CD had the least subvisible particles. In addition, DLS analysis showed no changes in aggregate size. Accelerated stability study results showed even better performance of HP-β-CD, with HMWP levels ranging from 2.60% to 2.64% over six months of storage, exceeding the performance of PS-20, PS-80, and P188. Discussion: The stabilization effect exerted by HP-β-CD could be assumed due to its capacity to decrease protein unfolding and prevent aggregation by the interfacial stress. Compared with the other surfactants, HP-β-CD provided better protection against mechanical and storage-related degradation of the protein while limiting particle formation. Conclusion: Under the conditions of agitation and storage stress, the use of HP-β-CD provides more efficient stabilization of recombinant human insulin as compared with polysorbate surfactants and Poloxamer 188.
INTRODUCTION/OBJECTIVE:The development of novel peptide-based therapeutics offers a promising strategy to overcome the limitations associated with conventional antibacterial drugs. This study aimed to design and synthesize a new series of lipidated pentapeptide derivatives containing unusual amino acids and to evaluate their antibacterial and antifungal potential. METHODS:A series of lipidated pentapeptides incorporating unconventional amino acids was designed and synthesized to enhance structural diversity, membrane interaction, amphiphilicity, and resistance to enzymatic degradation. Lipophilic moieties were introduced to improve peptide stability and biological performance. The synthesized compounds were characterized using standard analytical techniques and evaluated for their antibacterial and antifungal activities against selected Gram-positive and Gram-negative bacterial strains and pathogenic fungi. RESULTS:Several lipidated pentapeptide derivatives exhibited promising antibacterial activity against both Gram-positive and Gram-negative bacteria. In addition, notable antifungal activity was observed against pathogenic fungal strains. The results demonstrated that lipidation significantly enhanced the antimicrobial efficacy of the peptides. Among these peptide derivatives, compound 6a exhibited antibacterial activity against E. coli with MIC = 150 μg/mL, whereas compound 6b showed activity against S. aureus with MIC = 250 μg/mL. Compound 6c demonstrated the strongest antifungal activity against Candida albicans (MIC = 40 μg/mL), while compound 6e showed activity against Aspergillus niger (MIC = 80 μg/mL). DISCUSSION:The findings highlight peptide lipidation as an effective structural modification strategy to improve membrane interaction, bioactivity, and enzymatic stability of antimicrobial peptides. The incorporation of unusual amino acids further contributed to enhanced resistance against degradation, supporting their therapeutic potential. CONCLUSION:This study demonstrates that lipidated pentapeptide derivatives represent promising candidates for next-generation antimicrobial agents. Peptide lipidation emerges as a valuable approach for developing stable and potent peptide-based therapies to combat bacterial and fungal infections.
Abstract: Changes to the metabolic pathways of cancer cells present a plethora of possibilities for the therapeutic targeting of tumor cell metabolism. These alterations primarily involve the expression and regulation of proteins. Despite ongoing advancements and new targets, improving targeted drug delivery to cancer cells remains an area of active exploration. Mitochondria, the dynamic organelles central to cell survival and death, have become a promising target in cancer therapy due to their crucial role in cellular metabolism and metastasis. Differences between healthy and cancerous mitochondria, such as membrane potential, energy production, and genetic mutations, offer a unique opportunity for selective therapeutic targeting. Approaches to target the mitochondria of tumor cells have focused on metabolic inhibitors, photosensitizers, mitochondria-homing peptides, and functionalized nanoparticles. Compared to traditional drug delivery methods, peptide-functionalized nanoparticles provide greater stability, improved cellular uptake and retention, superior targeting, and remarkable flexibility in controlling the structure and function of designed conjugates. Peptide conjugation further enhances the biocompatibility and bioavailability of nanotherapeutics, delivering treatments that are not only more effective but also less toxic, and the ability to overcome drug resistance makes them a highly promising approach. This review highlights the recent progress in the development of mitochondria-homing peptides and peptide-functionalized nanoparticles, as well as peptide-based drug nanocarrier delivery methods, which show promising abilities to enhance the specific targeting and delivery of therapeutic drugs to cancer cells.
