Journal of Pharmacology and Experimental Therapeutics最新文献

Damage to the intestinal epithelial barrier is a hallmark of inflammatory diseases such as necrotizing enterocolitis. Specialized proresolving mediators (SPMs), such as lipoxin A4, resolvin D1, and resolvin E1, which are derived from essential fatty acids, have been shown to aid in resolving inflammation and promote mucosal healing. This study aimed to explore the effects of specific SPMs on intestinal inflammatory response in an early life in vitro model. We established 3-dimensional and 3-dimensional organoid cultures from fetal and pediatric intestines and investigated the effect of an SPM cocktail (lipoxin A4, resolvin D1, and resolvin E1) on gut epithelial maturation and barrier function. An inflammatory response of the gut barrier was provoked by lipopolysaccharide and flagellin stimulations combined with proinflammatory cytokines, tumor necrosis factor-α, and interferon gamma. Additionally, repetitive mechanical wounding was developed to test the effects of the SPM cocktail on 2-dimensional organoid monolayers. Under physiological conditions, we observed no effect of SPM cocktail treatment on gut epithelial maturation. Upon cytokine challenge, there was no modulation of the inflammatory tone of the gut barrier by the SPM cocktail. However, during the repetitive wounding and recovery assay, SPM cocktail treatment accelerated barrier recovery and maintained barrier integrity for 24 hours after repeated injuries. Our findings suggest that the SPM cocktail does not affect bacterial product- or cytokine-induced epithelial inflammation, although it may accelerate epithelial barrier recovery in mechanically wounded monolayers. These results provide valuable insights into the therapeutic potential of SPMs in neonatal intestinal inflammation. SIGNIFICANCE STATEMENT: Using early life intestinal organoid models, we found that although specialized proresolving mediators did not alter cytokine- or bacterial product-induced inflammation, they significantly enhanced epithelial barrier recovery following repeated mechanical injury.

Anaerobic bacteria induced colorectal cancer (CRC) represents a significant clinical concern. The understanding of cancer etiology has evolved significantly, from being predominantly viewed as genetically induced cancer to bacterial biofilm induced cancer. Despite the growing evidence linking bacterial virulence to tumor progression, the molecular interactions between bacterial biofilm proteins and anticancer drugs remain poorly understood. We explored the interaction of clinically used anticancer drugs (bevacizumab, capecitabine, fluorouracil, fruquintinib, leucovorin calcium, regorafenib, and tucatinib) with virulence proteins of oncomicrobes including Helicobacter pylori (cytotoxin-associated gene A), Fusobacterium nucleatum (Fusobacterium adhesion A), Bacteroides fragilis (Bfragilis toxin). Leucovorin calcium exhibited the highest binding affinity toward cytotoxin-associated gene A (-7.9 kcal/mol) through 7 hydrogen bonds. Similarly, regorafenib demonstrated strong interaction with Bfragilis toxin and Fusobacterium adhesion A, with binding affinities -8.6 and -6.5 kcal/mol, respectively, supported by multiple hydrogen and covalent bonds. Subsequent molecular dynamics simulations revealed low root mean square deviation and root mean square fluctuation values, indicating stable and compact drugs-protein interaction. Therefore, contributing to functional inactivation of bacterial virulence factors, thereby weakening bacterial colonization, biofilm formation, and events that sustain pro tumorigenic microenvironment. Overall, the present study provides computational evidence over anticancer drugs that may interact with bacterial virulence mechanisms implicated in anaerobic bacteria induced CRC, offering novel insights into therapeutic avenues capable of mitigating bacterial contributions in CRC initiation and progression. SIGNIFICANCE STATEMENT: The study focuses evaluating anticancer drugs targeting carcinogenic virulence proteins associated with bacterial biofilm mediated colorectal cancer.

Diabetic peripheral neuropathy (DPN) is a major complication of diabetes, characterized by progressive nerve damage and debilitating pain. Neuroinflammation plays a critical role in its pathogenesis, but therapeutic options remain limited. A disintegrin and metalloprotease 17 (ADAM17) regulates inflammatory signaling, but its ubiquitous expression makes it a difficult target. This study examined the role of inactive rhomboid protein 2 (iRhom2), a cofactor essential for ADAM17 activation, in the development of DPN. Diabetes was induced in wild-type (WT) and iRhom2 knockout (KO) mice using streptozotocin. Both groups developed hyperglycemia (>300 mg/dL); however, only WT mice exhibited significant mechanical and thermal hyposensitivity, characteristic of DPN. iRhom2 KO mice were protected from these deficits, suggesting a glucose-independent protective mechanism. In sciatic nerves of diabetic WT mice, expression of ADAM17, iRhom2, and tumor necrosis factor-α increased by 5.3-, 7.7-, and 48-fold, respectively; these changes were attenuated in KO mice. Histological analysis showed preservation of nerve fiber structure and reduced inflammatory infiltration in diabetic iRhom2 KOs. In cultured human microglial cells, high glucose triggered oxidative stress and induction of inflammatory mediators, including cyclooxygenase-2, interleukin-6, interleukin-8, tumor necrosis factor-α, and monocyte chemoattractant protein-1. Silencing of iRhom2 reduced these responses. These findings identify iRhom2 as a critical mediator of diabetic neuropathy, acting by regulating neuroinflammation. Deletion of iRhom2 confers glucose-independent protection against neuropathic pain, highlighting iRhom2 as a promising therapeutic target for preventing or treating DPN. SIGNIFICANCE STATEMENT: This study identifies iRhom2 as a key mediator of diabetic peripheral neuropathy by driving neuroinflammation and oxidative stress. Deletion of iRhom2 provided protection against neuropathic changes, without altering glucose levels, revealing a glucose-independent mechanism. These findings establish iRhom2 as a promising therapeutic target, offering new translational opportunities to prevent or treat diabetic neuropathy.

Therapeutic blockade of proinflammatory cytokines has revolutionized the treatment of rheumatoid arthritis (RA), leading to the approval of several therapeutic biologics for RA. A prominent target in RA is tumor necrosis factor-α (TNF-α), a proinflammatory cytokine. Etanercept (Enbrel), a fusion protein comprising the soluble portion of the p75-TNF receptor and the Fc fragment of human IgG1 (hinge, CH2, and CH3 domains) was the first TNF-α specific biologic to make a substantial impact for the treatment of RA. Enbrel (etanercept) differs structurally and functionally from other anti-TNF-α biologics (monoclonal antibodies), primarily because of its unique structure. This study aimed to explore whether structural modifications of Enbrel with specific focus on isotype variation and the incorporation of the CH1 domain to the Fc constant region, can potentiate its therapeutic efficacy. We developed 4 murine versions of Enbrel: mEnbrel2a and mEnbrel1, with and without the CH1 domain. These versions were assessed for their ability to bind and neutralize TNF-α in vitro, as well as their therapeutic effects in vivo using an experimental RA mouse model. We found that all mEnbrel variants bound TNF-α with comparable affinities. However, the mEnbrel2a derivatives, particularly with the CH1 domain, exhibited superior TNF-α neutralization in vitro. In vivo, mEnbrel2a with a CH1 domain provided the most significant reduction in disease severity. These findings underscore the critical role of isotype and domain selection in optimizing the therapeutic potential of Fc-fusion proteins and provide valuable insights applicable to other Fc-fusion proteins and a broader range of pathologies. SIGNIFICANCE STATEMENT: This study reveals that isotype and Fc domain engineering of tumor necrosis factor-α-targeting biologics enhances therapeutic efficacy against rheumatoid arthritis in a mouse model. Specifically, incorporation of the CH1 domain into an Enbrel-based Fc-fusion protein of the mouse IgG2a isotype significantly improved disease outcomes (delay of onset, arthritis severity, reduction in inflammatory white blood cells), highlighting the importance of Fc configuration for optimizing mEnbrel. These findings provide a foundation for the rational design of next-generation Fc-fusion therapeutics for autoimmune diseases.

Age-related macular degeneration (AMD) is a leading cause of permanent vision loss in older patients worldwide. The neovascular (wet) AMD is characterized by abnormal choroidal neovascularization driven by vascular endothelial growth factor (VEGF), platelet-derived growth factor, and Tie2 signaling pathways, leading to retinal damage and progressive vision decline. Current standard-of-care anti-VEGF therapies aim to limit choroidal neovascularization through extracellular targeting of cytokines involved in the VEGF signaling pathway implicated in angiogenesis. Although these existing therapies can be effective, many patients face a high treatment burden of multiple intraocular injections, which can negatively impact compliance, safety, and long-term efficacy. Tyrosine kinase inhibitors (TKIs) aim to address these limitations by offering longer durability, broad-spectrum targeting of angiogenic pathways, and a reduction in treatment burden through intracellular targeting of angiogenic pathways. With multiple pharmaceutical TKI candidates advancing through clinical trials and showing promising data, this class of drugs could lead to a shift in future treatment options for patients with wet AMD. Despite the progress TKIs have made, there have yet to be any candidates approved for wet AMD treatment. Much of the existing evidence is from early-phase and short-term studies, and questions remain about long-term efficacy and safety compared to current standard-of-care anti-VEGF therapies. Nevertheless, with multiple candidates advancing through phase III clinical trials, TKIs have the potential to emerge as a next-generation treatment class that may transform the wet AMD therapeutic landscape. SIGNIFICANCE STATEMENT: Given the chronic nature of wet age-related macular degeneration and the limitations of current anti-vascular endothelial growth factor therapies, tyrosine kinase inhibitors have emerged as a promising class of anti-angiogenic agents. This review highlights the recent clinical developments in this evolving therapeutic landscape.

Asthma-chronic obstructive pulmonary disease (COPD) overlap (ACO) is a chronic inflammatory airway condition that presents with features of both asthma and COPD, complicating its treatment and management. In this study, the potential of iguratimod-loaded nanostructured lipid carriers (IGU-NLCs) as an inhalable therapy for ACO was investigated. IGU-NLCs were formulated and characterized using Fourier-transform infrared, X-ray diffraction, thermogravimetric - differential scanning calorimetry, field emission scanning electron microscopy, and particle size analysis. The formulation demonstrated favorable physicochemical stability and nanoscale particle size distribution. IGU-NLCs demonstrated good cytocompatibility and minimal reactive oxygen species induction at 10 μg/mL in vitro, supporting their suitability for safe pulmonary delivery. An in vivo ACO model was induced by papain/cigarette smoke exposure, followed by treatment with plain IGU and IGU-NLCs via inhalation, to evaluate their therapeutic effects on oxidative stress, inflammation, and lung function. X-ray and ECG analyses revealed that IGU-NLCs more effectively reversed the airway obstruction and cardiac alterations induced by papain/cigarette smoke exposure. Histopathologic analysis showed significant improvement in lung architecture. Moreover, immunohistochemistry for CD68⁺ revealed reduced macrophage infiltration, indicating an anti-inflammatory effect. Overall, this study demonstrates that nebulized IGU-NLCs offer a noninvasive, targeted, and effective approach to mitigate ACO pathology, highlighting their potential for clinical translation in respiratory therapeutics. SIGNIFICANCE STATEMENT: Asthma-chronic obstructive pulmonary disease overlap lacks effective therapies because of its complex pathophysiology. This study repurposed iguratimod using nanostructured lipid carriers for inhalation, which reduced inflammation, oxidative stress, and lung damage in vivo, highlighting a novel, targeted strategy for this overlapped disease.

Methyl (1-{[6-{[(1S)-1-cyclopropylethyl]amino}-2-(pyrazolo[5,1-b][1,3]thiazol-7-yl)pyrimidin-4-yl]carbonyl}piperidin-4-yl)carbamate mono(4-methylbenzenesulfonate) monohydrate (NS-229) is a novel Janus kinase 1 inhibitor currently being evaluated in a phase 2 global study (NCT06046222) for the treatment of eosinophilic granulomatosis with polyangiitis (EGPA). We investigated the nonclinical efficacy of NS-229 to support its therapeutic use in treating EGPA. Its effects were investigated in human peripheral blood eosinophils, human peripheral blood mononuclear cells, and a mouse model of eosinophilic vasculitis induced by ovalbumin. In human peripheral blood eosinophils, NS-229 and an anti-interleukin (IL)-5 antibody, but not prednisolone, significantly decreased the expression of CD69 induced by IL-5. In human peripheral blood mononuclear cells, NS-229 and prednisolone, but not the anti-IL-5 antibody, significantly decreased the production of cytokines such as interferon gamma, IL-5, and IL-13, induced by anti-CD3/CD28 antibody. NS-229 inhibited the development of vascular lesions, decreased eosinophil counts in the blood and bronchoalveolar lavage fluid, and lowered bronchoalveolar lavage fluid lymphocyte counts in the ovalbumin-induced eosinophilic vasculitis mouse model. The effects of NS-229 in the mouse model were comparable to those of prednisolone and tofacitinib, a pan-Janus kinase inhibitor. Regarding safety, NS-229 did not influence the platelet or red blood cell counts, which were significantly elevated with tofacitinib and prednisolone, respectively. NS-229 did not affect body weight, which was significantly increased with tofacitinib and significantly decreased with prednisolone. Collectively, the nonclinical investigation of NS-229 showed a suppression of multiple cytokine signals and inhibition of vascular lesion formation without impacting the relevant side-effect parameters, suggesting its potential as an additional treatment option for EGPA. SIGNIFICANCE STATEMENT: NS-229 inhibited the formation of vascular lesions in a mouse model of ovalbumin-induced eosinophilic vasculitis without affecting certain side-effect parameters. The underlying mechanism of action is suggested to be the selective inhibition of multiple cytokine signals via JAK1.

Appropriate treatment of muscle spasticity and spasms is important as these conditions may significantly impair patients' quality of life. Conventional pharmacological treatments for these conditions have poor effectiveness and/or tolerability. Cannabis is being explored as a treatment. This was a longitudinal study of patient use of different cannabis products. Data was collected from patient surveys, clinic records, and changes in Patient Reported Outcome Measures Information System 29-Item scores over time. Patient-reported responses on health-related quality of life adverse events (n = 150) and outcomes (n = 78) from treatment for spasticity or spasms were analyzed. No improvements in physical functioning were observed for either group of patients across all product types. However, patients with spasticity who were using cannabidiol-only products experienced an improvement in sleep disturbance, fatigue, pain interference, and pain intensity. Patients with spasms who were using balanced, cannabidiol-dominant, or tetrahydrocannabinol-dominant products also experienced improvements in these 4 outcomes. Commonly reported adverse events were dry mouth, drowsiness, fatigue, dizziness, and nausea. Despite no observation of improvement in physical functioning, the results suggest that cannabis may help relieve some of the secondary complications associated with these conditions, such as poor sleep and pain. SIGNIFICANCE STATEMENT: This longitudinal study highlights differential benefits across cannabis product types, with cannabidiol-only formulations aiding spasticity-related symptoms and tetrahydrocannabinol- or cannabidiol-dominant products benefiting those with spasms. These findings support the potential of cannabis as a potential therapy to improve health-related quality of life in patients with limited options from conventional pharmacological treatments.