Journal of Pharmacology and Experimental Therapeutics最新文献

Type I conventional dendritic cells (cDC1s) are key drivers of antitumor immunity. In human cancers, their presence correlates with better prognosis and survival benefits. In preclinical mouse tumor models, cDC1s are indispensable for successful T-cell-mediated tumor killing and therapeutic response to immune checkpoint blockade therapies. The essential role of cDC1s in antitumor immunity stems from their ability to uptake tumor-derived antigens and traffic them to the tumor-draining lymph node (tdLN) for T-cell priming. At the tdLN, cDC1s present tumor antigens to naïve CD8⁺ T cells and polarize them into tumor-specific cytotoxic T cells that eventually kill the tumor cells. Within the tumor, cDC1s secrete the chemokines CXCL9 and CXCL10 that recruit CXCR3⁺ effector natural killer and T cells and thereby sustain a local cytotoxic T-cell response. The trafficking and migration of cDC1s to the tumor and tdLNs are largely mediated by a chemokine-chemokine receptor network linking cDC1s to their interacting immune cell partners. In this review, we discuss 2 key chemokine ligand-receptor pairs, C-C chemokine ligand 5-C-C chemokine receptor 5 and X-C chemokine ligand 1-X-C chemokine receptor 1, that play essential roles in directing cDC1 migration to the tumor. Strategies that harness these cDC1-recruiting chemokine systems offer invaluable therapeutic prospects for enhancing current vaccine design and cancer immunotherapies. SIGNIFICANCE STATEMENT: The lack of type I conventional dendritic cells (cDC1s) in tumors represents a major roadblock for current cancer immunotherapies. Here, we highlight 2 chemokines, C-C chemokine ligand 5 and X-C chemokine ligand 1, that are critical for recruiting cDC1s to the tumor microenvironment, where they uptake tumor antigens and cross-present antigens to T cells following migration to the lymph nodes. We further discuss recent advances and limitations in current dendritic cell vaccine design and cancer adjuvant therapies, and propose new strategies to enhance cDC1 recruitment into tumors.

Colorectal cancer remains a leading cause of cancer-related mortality, necessitating the development of novel therapeutic strategies. In this study, we investigate the pharmacological and toxicological properties of JAMC-4/108, a traditional Mongolian herbal extract, in human colonic adenocarcinoma (HT-29) and normal colonic epithelial (CCD 841 CoTr) cells. Using high-performance liquid chromatography-electrospray ionization-quadrupole time-of-flight-tandem mass spectrometry fingerprinting, we characterized its chemical composition, while a combination of cellular, biochemical and molecular assays provided mechanistic insights into its cytotoxic effects. The ethanol extract of JAMC-4/108 exhibited substantial selective cytotoxicity in HT-29 cells by inducing apoptosis and necrosis via caspase-3, cleaved caspase-7 and poly(ADP-ribose) polymerase activation, while the water extract primarily triggered caspase-9-mediated apoptosis. Both extracts modulated oxidative stress pathways, increasing Nrf2, Keap1 and LC3A/B levels, with the ethanol extract also upregulating NQO1, suggesting metabolic implications. In contrast, the ethanol extract had minimal apoptotic effects in normal colonic epithelial cells, whereas the water extract primarily influenced caspase-9 expression, indicating a distinct toxicity profile. Additionally, both extracts altered cell cycle progression-stimulated NOx release and modulated Fe³⁺ ion and DPPH radical pools in a concentration-dependent manner. Our findings highlight the selective anticancer potential of JAMC-4/108 and provide mechanistic insights into its pharmacological and toxicological effects, supporting its further evaluation for therapeutic development and safety assessment. SIGNIFICANCE STATEMENT: This study demonstrates the selective anticancer activity of JAMC-4/108, a Mongolian herbal extract, against colorectal cancer cells, with distinct apoptotic mechanisms and minimal toxicity to normal cells. These findings support the potential of JAMC-4/108 as a novel therapeutic candidate for colorectal cancer treatment, highlighting its pharmacological efficacy and safety profile for further development.

Metabolic dysfunction-associated steatohepatitis (MASH) poses a significant public health challenge, characterized by liver fat accumulation accompanied with inflammation and cell damage. Patients with MASH commonly exhibit hypercoagulability. Our previous work showed that direct oral anticoagulants exert anti-inflammatory effects in early stages of metabolic dysfunction. Mitigation of thromboinflammation ameliorated the manifestations of cardiometabolic complications. Here, we examine the protective effects of rivaroxaban in a rat model of MASH and compare it to representatives of 2 other antithrombotic drug classes, enoxaparin and clopidogrel, as well as silymarin, a bona fide hepatoprotective agent. Rats were divided into 6 groups: control, MASH (induced by an atherogenic diet), and treatment groups receiving either silymarin (50 mg/kg by mouth), rivaroxaban (20 mg/kg by mouth), enoxaparin (2 mg/kg subcutaneously), or clopidogrel (6.75 mg/kg by mouth) for 8 weeks, starting in the third week of induction. MASH rats showed elevated markers of visceral adipose thromboinflammation, along with liver injury markers (aspartate aminotransferase, alanine aminotransferase, and serum albumin), heightened hepatic levels of inflammatory cytokines (interleukin [IL]-1β and IL-6), reduced antioxidant capacity, and an imbalance in coagulation factors (elevated activated coagulation Factor X and Factor VIII/Protein C) compared with the control group. Anticoagulants and silymarin treatment led to varying degrees of amelioration of these MASH-associated abnormalities. Rivaroxaban demonstrated the most substantial improvement in thromboinflammatory markers, reaching levels comparable to the control group, with percentage improvements of approximately 52%, 49%, and 42% in activated coagulation Factor X, IL-1β, and IL-6, respectively. These findings suggest that direct oral anticoagulants hold promise as therapeutic agents for MASH by targeting the underlying thromboinflammatory state. SIGNIFICANCE STATEMENT: Metabolic dysfunction-associated steatohepatitis is a global health issue. In a rat model of the disease, rivaroxaban improves liver injury markers, outperforming bona fide hepatoprotective substances. This study emphasizes the role of thromboinflammation in metabolic dysfunction-associated steatohepatitis and highlights direct anticoagulants as a potential novel treatment approach.

The incidence of fatal drug overdoses has increased dramatically over the past decade due to the widespread availability of fentanyl and its analogs. As a complementary strategy to current overdose reversal agents, monoclonal antibodies (mAbs) are in development as therapeutics for prevention and reversal of fentanyl overdose. In the present study, the anti-fentanyl mAb HY6-F9 was tested for reversal of fentanyl-induced respiratory arrest (apnea) in a porcine model. In a first study, following fentanyl-induced apnea, chimeric HY6-F9 and naloxone control were administered as an intravenous bolus. Both chimeric HY6-F9 and naloxone restored spontaneous breathing within 90 seconds. Treatment with mAb increased the concentration of fentanyl in serum by 10-fold within the first minute after mAb bolus administration. In a second study, after induction of apnea, humanized HY6-F9 and naloxone control were administered as a slow intravenous infusion over 10 minutes to determine the ED₅₀ to restore baseline breathing. In this study, the mean ± SEM ED₅₀ of humanized HY6-F9 and naloxone to restore baseline respiratory rate were 16.0 ± 1.3 mg/kg and 6.9 ± 1.8 μg/kg, respectively. During mAb infusion, the concentration of fentanyl in serum increased proportionally to the concentration of infused mAb. The anti-fentanyl mAb ablated fentanyl-dependent opioid receptor activation in an in vitro system with concentrations of fentanyl similar to those observed in pigs after mAb treatment. These results demonstrate the efficacy of an anti-fentanyl mAb as a treatment to reverse fentanyl overdose. SIGNIFICANCE STATEMENT: Treatments for opioid use disorder and overdose are urgently needed. Here, we show that an anti-fentanyl monoclonal antibody reversed fentanyl-induced apnea in pigs, and caused rapid (<1 minute) redistribution of fentanyl into serum. Fentanyl was 99% bound by monoclonal antibodies and showed no activity at the opioid receptor.

Nondrug alternative reinforcers have long been used as a component of therapeutic interventions for the management of substance use disorder; however, the conditions under which alternative reinforcers are most effective are not well characterized. This study evaluated the impact of varying the magnitude of an alternative reinforcer on oxycodone self-administration and drug-primed responding in male and female squirrel monkeys. Subjects (n = 4/sex) were trained under concurrent second-order schedules of reinforcement for intravenous oxycodone (0.001-0.1 mg/kg per injection) on one lever and sweetened condensed milk (5%, 10%, 20%, and 30% in water) on another. Oxycodone-primed responding was evaluated by administering 0.32 mg/kg of oxycodone prior to sessions in which saline was available on the drug-paired lever. During oxycodone self-administration sessions, milk availability decreased oxycodone self-administration and preference in a concentration-dependent manner; low milk concentrations were more effective at decreasing oxycodone's reinforcing potency in males. During priming tests, milk significantly attenuated oxycodone-primed responding in both males and females; low milk concentrations were more effective at decreasing the priming effects of oxycodone in females. That alternative reinforcers differentially impacted self-administration and oxycodone-primed responding in a sex-dependent manner suggests that treatment strategies that use alternative reinforcers may be more effective in males or females depending on when they are implemented. SIGNIFICANCE STATEMENT: The use of nondrug reinforcer alternatives is a key component of many therapeutic interventions for substance use disorder. However, the variables that influence response to alternative reinforcers remain incompletely understood. This study investigated how varying the magnitude of an alternative reinforcer impacts oxycodone self-administration and drug-primed responding in male and female squirrel monkeys. Overall, the findings indicate that alternative reinforcers affect self-administration and oxycodone-primed responding differently depending on sex.

Triple-negative breast cancer (TNBC) is an aggressive subtype with poor prognosis and limited therapeutic options, lacking effective molecular targets. Recent studies have highlighted the critical role of mitochondrial metabolism, particularly oxidative phosphorylation, in TNBC progression. Mitochondrial RNA polymerase (POLRMT) has been implicated in maintaining mitochondrial function and stabilizing oxidative phosphorylation complexes. Targeting POLRMT offers a promising strategy to disrupt the heightened metabolic demands of tumors. Kaplan-Meier survival analysis revealed that elevated POLRMT levels were specifically associated with poor prognosis in TNBC patients. The Cancer Genome Atlas cohorts revealed that POLRMT transcripts are upregulated in TNBC tissues, and this enzyme overexpression was linked to hypomethylation of the cg23000464 site in the promoter region. We applied cell proliferation inhibition assays to investigate the potential anti-TNBC activity of targeted siRNA, an inhibitor of mitochondrial transcription (IMT) and mitochondrial protease targeting chimera (MtPTAC). Consistent with the silencing of POLRMT using siRNA both IMT and MtPTAC effectively suppress mitochondrial transcription, impacting oxidative phosphorylation, cell proliferation, and clonogenic ability in TNBC cells. In addition to eliciting comparable effects on cell phenotypes as IMT, MtPTAC selectively degrades POLRMT and inhibits the growth of IMT-resistant cells. POLRMT may represent a promising therapeutic target for TNBC, and MtPTAC could demonstrate promising application prospects. SIGNIFICANCE STATEMENT: This study shows that mitochondrial RNA polymerase (POLRMT) overexpression-linked to promoter hypomethylation-is associated with poor patient prognosis. Silencing or inhibiting POLRMT disrupts mitochondrial transcription, impairing oxidative phosphorylation, cell proliferation, and clonogenicity. Moreover, mitochondrial protease targeting chimera selectively degrades POLRMT and overcomes resistance to conventional inhibitors, underscoring its potential as an effective treatment option for aggressive triple-negative breast cancer.

Approximately 5% to 10% of asthma patients are resistant to glucocorticoid therapy; however, the mechanisms by which this resistance develops remain unclear. The present study investigated whether and how mechanistic target of rapamycin (mTOR) is involved in the development of steroid resistance using in vitro and in vivo murine models. The interleukin (IL)-33/thymic stromal lymphopoietin (TSLP)/IL-7-induced growth of group 2 innate lymphoid cells (ILC2) in vitro was resistant to dexamethasone (DEX), but suppressed by everolimus, an mTOR inhibitor, in a concentration-dependent manner. The inhibition of ILC2 growth by the combination of DEX and everolimus was significantly stronger than that by everolimus monotherapy. The combination of the pan-class I phosphatidylinositide-3 kinase inhibitor, buparlisib and the pan-Akt inhibitor, capivasertib also attenuated the resistance of IL-33/TSLP/IL-7-exposed ILC2s to DEX. The expression of the antiapoptotic factor, B-cell lymphoma-extra large, induced by IL-33/TSLP/IL-7 in ILC2s was significantly reduced by everolimus. Additionally, everolimus effectively suppressed the IL-33/TSLP/IL-7-induced phosphorylation of glucocorticoid receptors (GR) at the Ser234 residue, which has been reported to cause GR dysfunction. In the steroid-resistant asthma mouse model in vivo, under treatment with everolimus, DEX inhibited the development of airway remodeling and increased the number of ILC2s in the lungs. The present results suggest that the phosphoinositide 3-kinase/protein kinase B/mTOR pathway plays an essential role in the development of steroid resistance in ILC2s and asthma pathogenesis through both the expression of B-cell lymphoma-extra large and phosphorylation of GR. Therefore, mTOR inhibitors have the potential to restore glucocorticoid sensitivity and, thus, exert steroid-sparing effects in severe asthma patients. SIGNIFICANCE STATEMENT: Approximately 5% to 10% of asthma patients exhibit resistance to steroid therapy, posing a major clinical challenge due to limited treatment options. This study demonstrates that activation of the phosphoinositide 3-kinase/protein kinase B/mechanistic target of rapamycin (mTOR) pathway induces steroid resistance in group 2 innate lymphoid cells. Targeting mTOR with everolimus restores steroid sensitivity, highlighting mTOR inhibition as a promising pharmacotherapy for steroid-resistant asthma.

Graves' ophthalmopathy (GO) is an orbital inflammatory autoimmune disease with limited treatment options. Advances in understanding of disease pathogenesis, particularly the dysregulated insulin-like growth factor-1 receptor (IGF-1R) signaling network in the orbital fibroblasts, have led to the development of targeted therapies against IGF-1R for GO. In this study, we aimed to evaluate the preclinical therapeutic potential of CT102, an IGF-1R-targeting antisense oligonucleotide. Antisense oligonucleotides represent a promising class of therapeutics due to their direct regulation of disease-causing genes and their variants, providing a compelling alternative to traditional "protein-specific" therapies. A GO-related rat model was established via intraperitoneal injection of bovine thyroglobulin and validated via the elevated serum thyroid peroxidase antibodies and suppressed serum thyroid-stimulating hormone levels. The GO-related rat model exhibited stable and consistent pathologic alterations of the extraocular muscles. Ocular administration of CT102 is well tolerated, and high-dose CT102 treatment showed therapeutic benefits as illustrated by the downregulation of IGF-1R level in ocular muscle tissue and reduction in pathologic abnormalities. Restoration of GO-associated biomarkers, including serum thyroid peroxidase antibody and serum thyroid-stimulating hormone levels, was observed in the high-dose CT102 group compared with normal controls. Furthermore, CT102 demonstrates superior modulation of GO-associated biomarkers relative to 2 positive controls: teprotumumab, the only anti-IGF-1R antibody approved by the US Food and Drug Administration, and miR-143, an RNA therapeutic targeting IGF-1R. To our knowledge, this study provides, for the first time, a rationale for clinical trials of CT102 in patients with GO and highlights the potential of anti-IGF-1R antisense oligonucleotides as a therapeutic strategy for GO. SIGNIFICANCE STATEMENT: To our knowledge, this study is the first to describe the therapeutic potential of anti- insulin-like growth factor-1 receptor antisense oligonucleotides in Graves' ophthalmopathy (GO), providing the rationale for future clinical trials in patients with GO and highlighting the potential of anti-insulin-like growth factor-1 receptor antisense oligonucleotide as a therapeutic strategy for GO.

The pleiotropy of minocycline (MINO), including anti-inflammatory, antioxidant, antimigratory, anti-matrix metalloproteinase (MMP), and neuroprotective effects, has been extensively reported. A novel nonantibiotic MINO derivative, 10-butyl ether minocycline (BEM), was synthesized to retain the pleiotropy of MINO while minimizing side effects such as antibiotic resistance and gut dysbiosis. Previously, we showed that BEM reduced alcohol consumption in dependent murine and porcine models of alcohol use disorder. In this study, we investigated the molecular mechanisms of BEM to determine its potential as a therapeutic agent for neuroimmune and inflammatory conditions such as alcohol use disorder. Here, we report that BEM showed a nearly complete loss of antimicrobial activity against Escherichia coli, Salmonella typhi, and Candida albicans. BEM showed a dose-dependent reduction in cell viability as measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, similarly to MINO. BEM also suppressed lipopolysaccharide-induced microglial activation as shown by reduced Iba1 expression in immunohistochemistry and western blot analyses. Inhibition of MMP-9 by BEM (IC50 = 42.2 μM) was improved compared to MINO (IC50 = 60.3 μM), whereas MMP-8 inhibition was moderate (IC50: BEM = 69.4 μM; MINO = 45.4 μM). BEM was found to be effective in inhibiting vascular endothelial growth factor-induced endothelial cell migration and L-glutamine-induced reactive oxygen species levels. Limited inhibition of 15-lipoxygenase activity was observed (IC50: BEM = 92.6 μM; MINO = 65.6 μM). BEM was not toxic to mitochondria, even at high concentrations (200 μM). By eliminating antimicrobial properties while preserving therapeutic pleiotropy, BEM presents an advancement in the development of a promising candidate with multimodal mechanisms to treat neuroimmune-inflammatory pathologies. SIGNIFICANCE STATEMENT: We report mechanisms of action for butyl ether minocycline, a minocycline analog under evaluation for the treatment of alcohol use disorder, which may also show efficacy for other complex disease processes that involve inflammatory or neuroimmune components. We show that butyl ether minocycline had a nearly complete loss of antimicrobial action, yet retained the pleiotropy of minocycline, likely making it a better multimodal therapeutic for long-term treatment of complex diseases with neuroimmune-related components.

Chemical, biological, radiological, and nuclear (CBRN) hazards encompass CBRN materials that can have adverse effects if accidentally or deliberately released. With the increase in the use of CBRN-related agents for research, therapeutics, diagnostics, industrial use, etc., these agents have become available. Thus, the probability of occurrences of CBRN emergencies has also increased. Therefore, nations are striving hard to strengthen themselves to handle such situations. CBRN emergency preparedness is a vast area comprising devices, drugs, equipment, systems, etc. Each aspect of CBRN preparedness is humungous, and proportionately meager efforts are under progress for various reasons. The most important aspect is that it is a matter of state because the cost of CBRN preparedness is very high. Among the different aspects, drug development for the management of CBRN emergencies is extremely important wherein several efforts by the agencies have resulted in a handful of antidotes. Additionally, research toward the development of drugs and drug products for CBRN agents is an extremely important area to be encouraged. This paper highlights the challenges in CBRN drug development. Additionally, it also suggests the measures that may help to empower the researchers and manufacturers to contribute toward CBRN medical management preparedness. SIGNIFICANCE STATEMENT: This paper underscores the urgent need for focused drug development to address the unique challenges posed by chemical, biological, radiological, and nuclear threats. It highlights existing gaps and proposes strategies to strengthen medical preparedness and response.