Drug Development Research最新文献

The aim of this study was to establish a humanized immune system model in severe combined immunodeficient (SCID) mice, assess dendritic cell (DC) phenotype, and evaluate the therapeutic efficacy of a DC-based vaccine in a bladder cancer model. Bladder cancer was induced in SCID mice by injection of T24 cells, followed by human peripheral blood leukocyte (hu-PBL) inoculation to reconstitute the human immune system. DCs were generated in vitro by culturing hu-PBL for 5 days and matured on the eighth day. The DC vaccine was produced by coculturing with whole tumor antigen which was purified through freezing and melting T24 cells. The therapeutic efficacy of a DC vaccine was evaluated by administering the vaccine to SCID mice at Weeks 5 and 6 after T24 cell injection. Immune reconstitution, phenotype of DCs, tumor weight, and matrix metalloproteinase-7 (MMP-7) mRNA expression were assessed. All SCID mice successfully developed bladder cancer, confirmed as urothelial carcinoma. After hu-PBL inoculation, human IgG was detectable in mouse serum at Week 5, and spleen weight increased over time, indicating successful human immune system reconstitution. Phenotypic analysis of DCs showed high expression of maturation markers, including CD1a (78.07% ± 9.43%), CD80 (60.11% ± 20.50%), and CD83 (46.82% ± 14.15%), indicating functional and mature DCs. Therapeutic intervention with the DC vaccine significantly reduced tumor weight and MMP-7 mRNA expression, with statistical significance (p = 0.0004 for tumor weight and p = 0.0008 for MMP-7). This study successfully established a humanized immune system model in SCID mice and demonstrated that a DC-based vaccine effectively inhibits tumor growth in a bladder cancer model. These results support the potential of DC vaccines as a promising immunotherapeutic strategy for bladder cancer.

Herein, and based on the pharmacophoric features of doxorubicin (Dox); 133 steroids were screened to assess their ability to act as TOP II inhibitors for the discovery of those with promising anticancer activity. The cytotoxic inhibitory concentration 50 (IC₅₀) of the investigated steroids was determined against H1299, CaCo2, MDA-MB-468, and FaDu cancer cell lines and compared to Dox. Fluticasone propionate and fusidic acid exhibited the most potent antiproliferative effect against the MDA-MB-468 with IC₅₀ values of 10.4 ± 0.7 and 10.6 ± 1.7 μM, respectively. On the other hand, the outstanding antitumor members (beclomethasone dipropionate, fluticasone propionate, prednisolone, dexamethasone, and fusidic acid) were further investigated for their TOP II inhibitory potentials. Where the protein expression of TOP II was downregulated by 0.79, 0.76, and 0.67-fold change for fusidic acid, fluticasone propionate, and dexamethasone, respectively, compared to the control. Besides, the examined steroidal candidates were subjected to a molecular docking study towards the TOP II receptor in comparison to Dox and the co-crystallized ligand (EVP) as references. Moreover, molecular dynamics (MD) simulations were conducted on the aforementioned steroids along with Dox and EVP for 200 ns to validate the docking results. Consequently, steroids, in particular fusidic acid, fluticasone propionate, and dexamethasone, can be regarded as promising lead compounds targeting TOP II for cancer treatment along with their typical anti-inflammatory effects.

Osteosarcoma (OS) is a common malignant bone tumor, frequently associated with impaired osteogenic differentiation of tumor cells. Recent studies have suggested that the NOTCH signaling pathway plays a crucial role in maintaining tumor cell stemness and may influence their differentiation status. This study investigates the role of NOTCH2, a key receptor in the NOTCH family, in regulating osteogenic differentiation in OS. By analyzing public datasets, we compared the expression patterns and functional relevance of NOTCH1-4 in OS and identified NOTCH2 as the most significant. Using MG-63 and Saos-2 OS cell lines, we found that NOTCH2 silencing suppressed cell proliferation, invasion, and stem-like properties, while promoting osteogenic differentiation under inductive conditions. This was accompanied by increased expression of osteogenic markers. Further experiments demonstrated that Importazole, an Importin-β inhibitor, blocked the nuclear translocation of NOTCH2. Treatment with Importazole alone inhibited OS cell proliferation and invasion, reduced stem-like features, and enhanced osteogenic differentiation. When combined with NOTCH2 knockdown, Importazole exerted a synergistic effect, further inhibiting tumor progression and promoting differentiation. In vivo, xenograft models confirmed that the combination treatment more effectively suppressed tumor growth and induced osteoblast-like characteristics compared to either intervention alone. These findings indicate that NOTCH2 is a critical regulator of OS cell behavior, and that targeting NOTCH2 - especially in combination with Importazole - may offer a promising therapeutic strategy to promote differentiation and suppress tumor progression in OS.

Celiac disease (CD) is an autoimmune disorder which is triggered by gluten in genetically susceptible individuals. There is no successful therapy for CD. A strict gluten-free diet (GFD) is the only remedy used in clinical practice, which highlights the need to develop pharmacotherapeutic approaches to treat CD. This review discussed the data from genetic, biochemical, and immunological research, which has identified the mechanisms that causes activation of gluten which cause sequential immunological cascade through antigen presenting cell (APC) and human leukocyte antigen (HLA) dependent pathway. Recent studies aim to develop medications that stimulate repair of intestinal barrier, modify gluten peptides to make less immunogenic, regulate immune responses, and reduce CD associated symptoms. These approaches are mostly investigated in addition to GFD. In addition to these approaches, therapies that may work without gluten restriction need to be prioritized for patients who continue to experience symptoms despite strict adherence to GFD.

Mitigating myocardial ischemia-reperfusion (I/R) injury poses a significant challenge, necessitating the exploration of novel therapeutic targets. Sestrin2 (Sesn2), a stress-induced protein, has emerged as a potential candidate for attenuating I/R injury, yet its precise mechanisms remain elusive. The role of Sesn2 was investigated using an in vitro model of H9C2 cardiomyocytes subjected to hypoxia-reoxygenation (H/R). Sesn2 expression was modulated through overexpression techniques, and cellular responses, including cell viability, inflammatory factor production, mitochondrial function, oxidative stress, autophagy, and apoptosis, were assessed. Furthermore, the role of the Nrf2/SIRT3 signaling pathway in the mechanism was explored via treating cells with Nrf2 inhibitor ML385. Sesn2 overexpression significantly improved cell viability, attenuated inflammatory factor production, preserved mitochondrial function, and mitigated oxidative stress in H/R-exposed cardiomyocytes. Additionally, Sesn2 enhanced autophagy and modulated the Nrf2/SIRT3 signaling pathway. Moreover, Sesn2-mediated protection was reversed upon inhibition of Nrf2 signaling, underscoring the importance of this pathway in Sesn2-induced protection. Our findings may elucidate the mechanism of Sesn2-mediated protection and highlight its potential as a therapeutic target to ameliorate H/R-induced cardiomyocyte injury.

Botulinum neurotoxins are the most potent toxins responsible for causing flaccid paralysis of muscles by blocking the release of acetylcholine at the neuromuscular junction. There are no postexposure therapeutics and effective active/passive prophylaxis available for the treatment. Therefore, it is highly desirable to develop a potential antidote to counter botulinum neurotoxicity. In this study, ~800 molecules were mined by a structure similarity search from open databases and docked into the pocket of the catalytic domain of botulinum toxin type E using AutoDock 4.2. Twenty-four small molecules with the best scoring function were selected and evaluated using in vitro and in vivo assays. Among these, two molecules, NSC1011 and NSC1012, were identified as inhibiting the catalytic activity of BoNT/E, with IC₅₀ values of 31.25 ± 1.0 μM and 55.45 ± 5.2 μM and K_D of 5.54E−07 and 6.51E−06 M, respectively. To find inhibitors that can reverse the neurotoxicity more effectively, we have derived and synthesized 12 analogs of NSC1011. These compounds showed higher inhibition than the parent molecules, with IC₅₀ and K_D values of 4.375 ± 2.3 µM and 1.61E−08 M (C25.12) and 10.25 ± 3.0 µM and 4.70E-08 M (C25.9). Compounds C25.9 and C25.12 completely protected mice in premixed doses and led to significant extension in survival of up to 60 h with therapeutic treatment. This study showed that these 8-HQ derivatives had the potency to inhibit BoNT/E by interacting with the active site. Further studies could lead to the development of undiscovered postexposure therapeutics against this deadly neurotoxin.

The growing resistance of malarial parasites to antiplasmodial drugs has necessitated the development of a new class of potent molecules to reduce the global malaria burden. This study evaluated the antimalarial efficacy of the cationic lipid stearylamine (SA) on blood-stage Plasmodium falciparum using SYBR-Green I assay. We conducted in vitro studies to assess the timing of SA action in drug-sensitive (Pf3D7) and chloroquine-resistant (PfINDO) strains of P. falciparum. Notably, SA demonstrated fast-acting cytostatic potential with 50% inhibitory concentration (IC₅₀s) values of 2.17, 1.97, and 1.33 µg/mL for Pf3D7 and 3.50, 2.76, and 1.94 µg/mL for PfINDO during the first generation cycle (12, 24, and 48 h), and the activity was maintained during the second generation cycle with IC₅₀ values at 72 h (1.15 µg/mL on Pf3D7 and 1.65 µg/mL on PfINDO) and 96 h (1.43 µg/mL on Pf3D7 and 1.10 µg/mL on PfINDO). Additionally, we explored the cytocidal potential of SA by exposing the parasites for 1, 2, 4, and 6 h and subsequent incubation for 48 h in SA-free conditions, which revealed an average IC₅₀ value of 1.68 µg/mL, demonstrating its irreversible parasite growth arrest. Moreover, SA induced reactive oxygen species (ROS) production at IC₅₀ concentrations, with minimal hemolytic effects. Our findings indicated that incorporating SA into lipid vehicles or other delivery systems loaded with antimalarial drugs can significantly reduce drug toxicity, enhance efficacy, and slow clinical resistance. Nevertheless, further preclinical studies are warranted to advance the antimalarial drug discovery pipeline.

Melanoma is a type of aggressive cancer distinguished by its high propensity for recurrence, the development of metastases, and an unfavorable outlook for recovery. Treatment modalities for melanoma encompass surgery, immunotherapy, and targeted therapies. In recent decades, berberine has garnered attention for its significant anticancer properties across various cancer types. This review systematically examines the molecular mechanisms of berberine in melanoma, particularly its modulation of critical signaling pathways, including B-RAF/MEK/ERK, PI3K/AKT, and NF-κB, which are essential for regulating melanoma cell proliferation and promoting apoptosis. Furthermore, berberine activates AMP-activated protein kinase, leading to the inhibition of cyclooxygenase-2, thereby reducing melanoma cell migration and invasion through decreased inflammation and enhanced cellular energy regulation. It also induces mitochondrial dysfunction and oxidative stress, promoting apoptosis while simultaneously inhibiting epithelial-to-mesenchymal transition, a key process in metastasis. Additionally, berberine modulates the immune microenvironment through Toll-like receptors, cytokine networks, and the regulation of various immune cells, thereby enhancing its antitumor effects. Recent studies have shown that the therapeutic effect of berberine is enhanced when used in combination with other therapies, especially immune checkpoint inhibitors, to improve antitumor immune responses. These findings highlight the potential of berberine as a multi-targeted agent for the treatment of melanoma, providing an avenue for further clinical exploration and integration into therapeutic strategies.

The space environment, characterized by microgravity and elevated radiation, offers a unique platform for scientific research with transformative potential for biomedical and pharmaceutical industries. As launch costs have decreased and commercial innovation has advanced, utilization of space for research has surged, with both space stations and nano/microsatellites (CubeSats) serving as essential platforms for ground breaking experiments. This systematic review summarizing findings from 86 peer-reviewed articles and major space research initiatives, focusing on the biological and medical insights gained from space-based investigations. Studies conducted in microgravity have revealed significant alterations in bacterial physiology, including increased virulence and antibiotic resistance, as well as enhanced secondary metabolite production with potential pharmaceutical applications. Human physiological changes, such as muscle atrophy, bone demineralization, and cardiovascular deconditioning, mirror accelerated aging and disease states, providing valuable models for understanding and developing treatments for similar conditions on Earth. Space research has also highlighted the risk of kidney stone formation due to altered calcium metabolism and gut microbiome shifts, along with ophthalmological abnormalities such as Spaceflight-Associated Neuro-Ocular Syndrome (SANS), which offer insights into terrestrial eye diseases. Advanced technologies, including 3D bioprinting, lab-on-a-chip, and tissue chips, have enabled sophisticated experiments in regenerative medicine and disease modeling. Microgravity facilitates the growth of high-quality drug crystals, improving drug stability, efficacy, and delivery methods, as exemplified by innovations in monoclonal antibody formulations and cancer therapeutics. Despite these advances, challenges such as limited data availability, high operational costs, and the complexity of translating space findings to Earth-based applications remain. In conclusion, space-based research is driving significant advancements in pharmaceutical science and medicine, uncovering novel disease mechanisms, therapeutic targets, and drug development strategies. Continued investment and interdisciplinary collaboration are essential to realize the full potential of space research for global healthcare innovation.

In this study, we aimed to design and synthesize a novel series of pyridine and pyrimidine derivatives and evaluate their anti-inflammatory activity against RAW 264.7 macrophages. Using chalcones (5a−f) as suitable precursors, we disclosed a novel series of pyridine (7a−f) and pyrimidine (9a−e) derivatives via the reaction of 5a−f with 2-cyanothioacetamide or guanidine hydrochloride, respectively. Both pyridines and pyrimidines were tested as anti-inflammatory agents to compare the difference in activity of the pyridine and pyrimidine scaffolds as part of a comparative study. With a percentage of live cells greater than 80%, the pyridines (7a−f) and pyrimidines (9a−e) were found to be safe for RAW cells. Moreover, the anti-inflammatory activity of these compounds was evaluated in lipopolysaccharide (LPS)-stimulated RAW macrophages by performing nitric oxide (NO) assays. Among pyridines, 7a and 7f showed significant inhibition with 65.48% and 51.19%, with IC₅₀ values (IC₅₀ = 76.6 and 96.8 µM), respectively. The pyrimidine derivatives showed promising results as well, 9a and 9d ranking the best activity with 55.95% and 61.90%, respectively, and IC₅₀ values (IC₅₀ = 83.1 and 88.7 µM, respectively). The gene expression levels were assessed for the most promising compounds 7a and 9d using real-time reverse transcription-polymerase chain reaction analysis to measure the mRNA and protein expression levels of inflammatory cytokines, including interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-ɑ), nuclear factor kappa β (NF-kβ), and inducible nitric oxide synthase (INOS). The expression levels of IL-1, IL-6, TNF-ɑ, NF-kβ, and INOS genes were decreased significantly in RAW-treated cells with 7a by 43%, 32%, 61%, 26%, and 53% respectively, compared with negative RAW cells. The expression levels of IL-1, IL-6, NF-kβ, and INOS genes were decreased significantly in RAW-treated cells with 9d by 71%, 48%, 61%, and 65%, respectively, compared with negative RAW cells. However, the expression levels of the TNF-ɑ gene were decreased without significant differences in RAW treated with 9d by 83% (p > 0.05) compared with negative RAW cells. These findings exhibited that 7a was more effective compared with 9d as an anti-inflammatory agent.