RÉSUMÉ
Botrytis cinerea is a notorious pathogen causing pre- and post-harvest spoilage in many economically important crops. Excessive application of site-specific fungicides to control the pathogen has led to the selection of strains possessing target site alterations associated with resistance to these fungicides and/or strains overexpressing efflux transporters associated with multidrug resistance (MDR). MDR in B. cinerea has been correlated with the overexpression of atrB and mfsM2, encoding an ATP-binding cassette (ABC) and a major facilitator superfamily (MFS) transporter, respectively. However, it remains unknown whether other transporters may also contribute to the MDR phenotype. In the current study, the transcriptome of a B. cinerea multidrug-resistant (MDR) field strain was analysed upon exposure to the fungicide fludioxonil, and compared to the B05.10 reference strain. The transcriptome of this field strain displayed significant differences as compared to B05.10, including genes involved in sugar membrane transport, toxin production and virulence. Among the induced genes in the field strain, even before exposure to fludioxonil, were several putatively encoding ABC and MFS transmembrane transporters. Overexpression of a highly induced MFS transporter gene in the B05.10 strain led to an increased tolerance to the fungicides fluopyram and boscalid, indicating an involvement in efflux transport of these compounds. Overall, the data from this study give insights towards better understanding the molecular mechanisms involved in MDR and fitness cost, contributing to the development of more efficient control strategies against this pathogen.
Sujet(s)
Botrytis , Dioxoles , Fongicides industriels , Transcriptome , Botrytis/effets des médicaments et des substances chimiques , Botrytis/génétique , Botrytis/pathogénicité , Transcriptome/génétique , Fongicides industriels/pharmacologie , Dioxoles/pharmacologie , Pyrroles/pharmacologie , Régulation de l'expression des gènes fongiques/effets des médicaments et des substances chimiques , Protéines fongiques/métabolisme , Protéines fongiques/génétique , Analyse de profil d'expression de gènes , Multirésistance des champignons aux médicaments/génétique , Résistance des champignons aux médicaments/génétique , Résistance des champignons aux médicaments/effets des médicaments et des substances chimiques , Aptitude génétiqueRÉSUMÉ
Corticotropin-releasing factor (CRF) is a key neuropeptide hormone that is secreted from the hypothalamus. It is the master hormone of the HPA axis, which orchestrates the physiological and behavioral responses to stress. Many disorders, including anxiety, depression, addiction relapse, and others, are related to over-activation of this system. Thus, new molecules that may interfere with CRF receptor binding may be of value to treat neuropsychiatric stress-related disorders. Also, CRF1R antagonists have recently emerged as potential treatment options for congenital adrenal hyperplasia. Previously, several series of CRF1 receptor antagonists were developed by our group. In continuation of our efforts in this direction, herein we report the synthesis and biological evaluation of a new series of CRF1R antagonists. Representative compounds were evaluated for their binding affinities compared to antalarmin. Four compounds (2, 5, 20, and 21) showed log IC50 values of -8.22, -7.95, -8.04, and -7.88, respectively, compared to -7.78 for antalarmin. This result indicates that these four compounds are superior to antalarmin by 2.5, 1.4, 1.7, and 1.25 times, respectively. It is worth mentioning that compound 2, in terms of IC50, is among the best CRF1R antagonists ever developed in the last 40 years. The in silico physicochemical properties of the lead compounds showed good drug-like properties. Thus, further research in this direction may lead to better and safer CRF receptor antagonists that may have clinical applications, particularly for stress-related disorders and the treatment of congenital adrenal hyperplasia.
Sujet(s)
Hyperplasie congénitale des surrénales , Conception de médicament , Pyrimidines , Récepteur CRH , Récepteur CRH/antagonistes et inhibiteurs , Récepteur CRH/métabolisme , Pyrimidines/composition chimique , Pyrimidines/pharmacologie , Pyrimidines/synthèse chimique , Humains , Hyperplasie congénitale des surrénales/traitement médicamenteux , Hyperplasie congénitale des surrénales/métabolisme , Pyrroles/composition chimique , Pyrroles/synthèse chimique , Pyrroles/pharmacologie , Corticolibérine/métabolisme , Stress psychologique/traitement médicamenteux , Simulation de docking moléculaireRÉSUMÉ
Since the largest and most fatal Ebola virus epidemic during 2014-2016, there have been several consecutive filoviral outbreaks in recent years, including those in 2021, 2022, and 2023. Ongoing outbreak prevalence and limited FDA-approved filoviral therapeutics emphasize the need for novel small molecule treatments. Here, we showcase the structure-activity relationship development of N-substituted pyrrole-based heterocycles and their potent, submicromolar entry inhibition against diverse filoviruses in a target-based pseudovirus assay. Inhibitor antiviral activity was validated using replication-competent Ebola, Sudan, and Marburg viruses. Mutational analysis was used to map the targeted region within the Ebola virus glycoprotein. Antiviral counter-screen and phospholipidosis assays were performed to demonstrate the reduced off-target activity of these filoviral entry inhibitors. Favorable antiviral potency, selectivity, and drug-like properties of the N-substituted pyrrole-based heterocycles support their potential as broad-spectrum antifiloviral treatments.
Sujet(s)
Antiviraux , Ebolavirus , Pyrroles , Pénétration virale , Pyrroles/pharmacologie , Pyrroles/composition chimique , Pyrroles/synthèse chimique , Antiviraux/pharmacologie , Antiviraux/composition chimique , Antiviraux/synthèse chimique , Humains , Relation structure-activité , Ebolavirus/effets des médicaments et des substances chimiques , Pénétration virale/effets des médicaments et des substances chimiques , Composés hétérocycliques/pharmacologie , Composés hétérocycliques/composition chimique , Composés hétérocycliques/synthèse chimique , Filoviridae/effets des médicaments et des substances chimiques , Marburgvirus/effets des médicaments et des substances chimiquesRÉSUMÉ
BACKGROUND: Vonoprazan, a potassium-competitive acid blocker, is superior to traditional proton pump inhibitor (PPI) in acid suppression and has been approved in the treatment of acid-related disorders. Accumulating evidence suggest associations between PPI use and gut microbiota, yet the effect of vonoprazan on GI microbiota is obscure. METHODS: Transgenic FVB/N insulin-gastrin (INS-GAS) mice as a model of gastric cancer (GC) were administered vonoprazan by gavage every other day for 12 weeks. Stomachs were evaluated by histopathology, Ki-67 proliferation index, and inflammatory cytokines. The mucosal and lumen microbiota from stomach, jejunum, ileum, cecum, and feces were detected using 16S rRNA gene sequencing. RESULTS: Higher incidence of intestinal metaplasia and epithelial proliferation were observed in the vonoprazan group than that in the control mice. Vonoprazan also elevated the gastric expression of proinflammatory cytokines, including TNF-α, IL-1ß, and IL-6. Each mice comprised a unique microbiota composition that was consistent across different niches. The structure of GI microbiota changed dramatically after vonoprazan treatment with the stomach being the most disturbed segment. Vonoprazan administration shifted the gut microbiota toward the enrichment of pathogenic Streptococcus, Staphylococcus, Bilophila, and the loss of commensal Prevotella, Bifidobacterium, and Faecalibacterium. Interestingly, compared to the controls, microbial interactions were weaker in the stomach while stronger in the jejunum of the vonoprazan group. CONCLUSIONS: Long-term vonoprazan treatment promoted gastric lesions in male INS-GAS mice, with the disequilibrium of GI microbiome. The clinical application of vonoprazan needs to be judicious particularly among those with high risk of GC.
Sujet(s)
Microbiome gastro-intestinal , Pyrroles , Tumeurs de l'estomac , Sulfonamides , Animaux , Pyrroles/administration et posologie , Pyrroles/pharmacologie , Sulfonamides/administration et posologie , Sulfonamides/pharmacologie , Microbiome gastro-intestinal/effets des médicaments et des substances chimiques , Tumeurs de l'estomac/microbiologie , Tumeurs de l'estomac/anatomopathologie , Souris , Souris transgéniques , ARN ribosomique 16S/génétique , Modèles animaux de maladie humaine , Mâle , Inhibiteurs de la pompe à protons/effets indésirables , Inhibiteurs de la pompe à protons/administration et posologie , Cytokines/métabolismeRÉSUMÉ
Pulmonary hypertension (PH) arises from increased pulmonary vascular resistance due to contraction and remodeling of the pulmonary arteries. The structural changes include thickening of the smooth muscle layer from increased proliferation and resistance to apoptosis. The mechanisms underlying apoptosis resistance in PH are not fully understood. In cancer cells, high expression of aquaporin 1 (AQP1), a water channel, is associated with apoptosis resistance. We showed AQP1 protein was expressed in pulmonary arterial smooth muscle cells (PASMCs) and upregulated in preclinical PH models. In this study, we used PASMCs isolated from control male rats and the SU5416 plus hypoxia (SuHx) model to test the role of AQP1 in modulating susceptibility to apoptosis. We found the elevated level of AQP1 in PASMCs from SuHx rats was necessary for resistance to apoptosis and that apoptosis resistance could be conferred by increasing AQP1 in control PASMCs. In exploring the downstream pathways involved, we found AQP1 levels influence the expression of Bcl-2, with enhanced AQP1 levels corresponding to increased Bcl-2 expression, reducing the ratio of BAX to Bcl-2, consistent with apoptosis resistance. These results provide a mechanism by which AQP1 can regulate PASMC fate.
Sujet(s)
Apoptose , Aquaporine-1 , Hypoxie , Indoles , Muscles lisses vasculaires , Myocytes du muscle lisse , Artère pulmonaire , Pyrroles , Animaux , Aquaporine-1/métabolisme , Aquaporine-1/génétique , Mâle , Artère pulmonaire/métabolisme , Artère pulmonaire/anatomopathologie , Artère pulmonaire/cytologie , Rats , Myocytes du muscle lisse/métabolisme , Myocytes du muscle lisse/anatomopathologie , Muscles lisses vasculaires/métabolisme , Muscles lisses vasculaires/anatomopathologie , Muscles lisses vasculaires/cytologie , Pyrroles/pharmacologie , Indoles/pharmacologie , Hypoxie/métabolisme , Rat Sprague-Dawley , Hypertension pulmonaire/métabolisme , Hypertension pulmonaire/anatomopathologie , Cellules cultivées , Protéines proto-oncogènes c-bcl-2/métabolisme , Protéines proto-oncogènes c-bcl-2/génétique , Modèles animaux de maladie humaineRÉSUMÉ
A 12-year-old male domestic cat with multiple subcutaneous mast cell tumours (MCTs) presented with a 2-week history of pruritus and raw/bleeding skin from self-trauma at Kagoshima University Veterinary Teaching Hospital. Polymerase chain reaction (PCR) and histopathological analyses revealed intertumoral heterogeneity among tumour locations based on the mutation status of KIT. In addition, the expression pattern of KIT was characterized. After failed treatment with vinblastine (2.0-2.2 mg/m2, intravenous administration, two doses in total) or nimustine (25 mg/m2, intravenous administration, two doses in total), toceranib (2.2-2.6 mg/kg, orally administered, every other day) was administered to treat recurrent MCTs harbouring the KIT exon eight internal tandem duplication mutation, achieving a complete response. However, toceranib resistance developed 2 months after treatment initiation. Subsequent PCR analysis was conducted to identify the mutational status of KIT in each MCT and to detect the presence of secondary mutations associated with the acquisition of toceranib resistance. Secondary KIT mutations (c.998G>C and c.2383G>C), which were not initially detected in tumour cells at diagnosis, were identified after the development of resistance to toceranib. This indicates that the tumour cells in feline MCTs in the same case have diverse characteristics. Our findings encourage further investigation into the development of therapeutic strategies for feline MCTs, particularly focusing on the heterogeneous nature of KIT/KIT and overcoming acquired resistance to toceranib.
Sujet(s)
Maladies des chats , Résistance aux médicaments antinéoplasiques , Indoles , Mutation , Protéines proto-oncogènes c-kit , Pyrroles , Animaux , Mâle , Chats , Maladies des chats/traitement médicamenteux , Maladies des chats/génétique , Indoles/pharmacologie , Indoles/usage thérapeutique , Pyrroles/pharmacologie , Pyrroles/usage thérapeutique , Protéines proto-oncogènes c-kit/génétique , Résistance aux médicaments antinéoplasiques/génétique , Antinéoplasiques/usage thérapeutique , Antinéoplasiques/pharmacologieRÉSUMÉ
Microglia, traditionally regarded as innate immune cells in the brain, drive neuroinflammation and synaptic dysfunctions in the early phases of Alzheimer disease (AD), acting upstream to Aß accumulation. Colony stimulating factor 1-receptor (CSF-1R) is predominantly expressed on microglia and its levels are significantly increased in neurodegenerative diseases, possibly contributing to the chronic inflammatory microglial response. On the other hand, CSF-1R inhibitors confer neuroprotection in preclinical models of neurodegenerative diseases. Here, we determined the effects of the CSF-1R inhibitor PLX3397 on the Aß-mediated synaptic alterations in ex vivo hippocampal slices. Electrophysiological findings show that PLX3397 rescues LTP impairment and neurotransmission changes induced by Aß. In addition, using confocal imaging experiments, we demonstrate that PLX3397 stimulates a microglial transition toward a phagocytic phenotype, which in turn promotes the clearance of Aß from glutamatergic terminals. We believe that the selective pruning of Aß-loaded synaptic terminals might contribute to the restoration of LTP and excitatory transmission alterations observed upon acute PLX3397 treatment. This result is in accordance with the mechanism proposed for CSF1R inhibitors, that is to eliminate responsive microglia and replace it with newly generated, homeostatic microglia, capable of promoting brain repair. Overall, our findings identify a connection between the rapid microglia adjustments and the early synaptic alterations observed in AD, possibly highlighting a novel disease-modifying target.
Sujet(s)
Aminopyridines , Peptides bêta-amyloïdes , Hippocampe , Potentialisation à long terme , Microglie , Animaux , Microglie/effets des médicaments et des substances chimiques , Microglie/métabolisme , Hippocampe/effets des médicaments et des substances chimiques , Hippocampe/métabolisme , Peptides bêta-amyloïdes/métabolisme , Potentialisation à long terme/effets des médicaments et des substances chimiques , Mâle , Aminopyridines/pharmacologie , Récepteur de facteur de croissance granulocyte-macrophage/antagonistes et inhibiteurs , Récepteur de facteur de croissance granulocyte-macrophage/métabolisme , Pyrroles/pharmacologie , Souris , Phagocytose/effets des médicaments et des substances chimiques , Transmission synaptique/effets des médicaments et des substances chimiques , Souris de lignée C57BL , Maladie d'Alzheimer/métabolisme , Maladie d'Alzheimer/physiopathologie , Acide glutamique/métabolismeRÉSUMÉ
Rosacea is a chronic inflammatory skin disease characterized by facial erythema and telangiectasia. Despite ongoing research, the pathogenesis of rosacea remains incompletely understood, and current therapies are not entirely satisfactory. The JAK/STAT signaling pathway plays an essential role in immunoregulation, inflammation, and neurovascular regulation. Inhibition of the JAK/STAT pathway appears to hold promise as a potential therapy for rosacea. This study aimed to investigate the effects of the JAK inhibitor tofacitinib on rosacea and to preliminarily explore its therapeutic mechanism. To this end, a rosacea-like mouse model was induced using LL37 and treated with a 2% tofacitinib emulsion. The results demonstrated that topical application of tofacitinib significantly ameliorated rosacea-like phenotype, reduced the infiltration of CD4+ T cells and mast cells, and suppressed dermal angiogenesis. RT-qPCR analysis revealed a reduction in mRNA expression levels of STAT1, STAT4, and STAT5a in skin lesions following topical tofacitinib treatment. Additionally, three patients diagnosed with erythematotelangiectatic rosacea (ETR) were included in the study and treated with oral tofacitinib, leading to a significant improvement in erythema and flushing symptoms. These findings collectively suggest that tofacitinib alleviates LL37-induced rosacea-like skin inflammation in mice and rosacea skin lesions by inhibiting the JAK/STAT signaling pathway.
Sujet(s)
Pipéridines , Pyrimidines , Rosacée , Transduction du signal , Adulte , Animaux , Femelle , Humains , Mâle , Souris , Adulte d'âge moyen , Administration par voie orale , Administration par voie topique , Lymphocytes T CD4+/effets des médicaments et des substances chimiques , Lymphocytes T CD4+/immunologie , Modèles animaux de maladie humaine , Inhibiteurs des Janus kinases/pharmacologie , Inhibiteurs des Janus kinases/usage thérapeutique , Inhibiteurs des Janus kinases/administration et posologie , Janus kinases/métabolisme , Janus kinases/antagonistes et inhibiteurs , Mastocytes/effets des médicaments et des substances chimiques , Mastocytes/immunologie , Mastocytes/métabolisme , Pipéridines/pharmacologie , Pipéridines/usage thérapeutique , Pipéridines/administration et posologie , Pyrimidines/pharmacologie , Pyrimidines/administration et posologie , Pyrimidines/usage thérapeutique , Pyrroles/pharmacologie , Pyrroles/administration et posologie , Rosacée/traitement médicamenteux , Transduction du signal/effets des médicaments et des substances chimiques , Peau/anatomopathologie , Peau/effets des médicaments et des substances chimiques , Peau/métabolisme , Facteurs de transcription STAT/métabolisme , Facteurs de transcription STAT/antagonistes et inhibiteurs , Facteur de transcription STAT-1/métabolismeRÉSUMÉ
Reduction in muscle contractile force associated with many clinical conditions incurs serious morbidity and increased mortality. Here, we report the first evidence that JAK inhibition impacts contractile force in normal human muscle. Muscle biopsies were taken from patients who were randomized to receive tofacitinib (n = 16) or placebo (n = 17) for 48 h. Single-fiber contractile force and molecular studies were carried out. The contractile force of individual diaphragm myofibers pooled from the tofacitinib group (n = 248 fibers) was significantly higher than those from the placebo group (n = 238 fibers), with a 15.7% greater mean maximum specific force (P = 0.0016). Tofacitinib treatment similarly increased fiber force in the serratus anterior muscle. The increased force was associated with reduced muscle protein oxidation and FoxO-ubiquitination-proteasome signaling, and increased levels of smooth muscle MYLK. Inhibition of MYLK attenuated the tofacitinib-dependent increase in fiber force. These data demonstrate that tofacitinib increases the contractile force of skeletal muscle and offers several underlying mechanisms. Inhibition of the JAK-STAT pathway is thus a potential new therapy for the muscle dysfunction that occurs in many clinical conditions.
Sujet(s)
Inhibiteurs des Janus kinases , Contraction musculaire , Muscles squelettiques , Pipéridines , Pyrimidines , Humains , Pipéridines/pharmacologie , Pyrimidines/pharmacologie , Contraction musculaire/effets des médicaments et des substances chimiques , Muscles squelettiques/effets des médicaments et des substances chimiques , Muscles squelettiques/métabolisme , Inhibiteurs des Janus kinases/pharmacologie , Mâle , Pyrroles/pharmacologie , Femelle , Adulte , Transduction du signal/effets des médicaments et des substances chimiques , Adulte d'âge moyen , Janus kinases/métabolisme , Fibres musculaires squelettiques/effets des médicaments et des substances chimiques , Fibres musculaires squelettiques/métabolismeRÉSUMÉ
Fludioxonil, an antifungal agent used as a pesticide, leaves a measurable residue in fruits and vegetables. It has been identified to cause endocrine disruption, interrupt normal development, and cause various diseases such as cancers. In this study, fludioxonil was examined for its effects on the development and metastasis of breast cancer cells. On fludioxonil exposure (10-5 M) for 72 h, mutant p53 (mutp53) MDA-MB-231 triple-negative breast cancer (TNBC) cells significantly inhibited cell viability and developed into polyploid giant cancer cells (PGCCs), with an increase in the number of nuclei and expansion in the cell body size. Fludioxonil exposure disrupted the normal cell cycle phase ratio, resulting in a new peak. In addition, PGCCs showed greater motility than the control and were resistant to anticancer drugs, i.e., doxorubicin, cisplatin, and 5-fluorouracil. Cyclin E1, nuclear factor kappa B (NF-κB), and p53 expressions were remarkably increased, and the expression of cell cycle-, epithelial-mesenchymal-transition (EMT)-, and cancer stemness-related proteins were increased in the PGCCs. The daughter cells obtained from PGCCs had the single nucleus but maintained their enlarged cell size and showed greater cell migration ability and resistance to the anticancer agents. Consequently, fludioxonil accumulated Cyclin E1 and promoted the inflammatory cytokine-enriched microenvironment through the up-regulation of TNF and NF-κB which led to the transformation to PGCCs via abnormal cell cycles such as mitotic delay and mitotic slippage in mutp53 TNBC MDA-MB-231 cells. PGCCs and their daughter cells exhibited significant migration ability, chemo-resistance, and cancer stemness. These results strongly suggest that fludioxonil, as an inducer of potential genotoxicity, may induce the formation of PGCCs, leading to the formation of metastatic and stem cell-like breast cancer cells.
Sujet(s)
Dioxoles , Transition épithélio-mésenchymateuse , Cellules souches tumorales , Polyploïdie , Pyrroles , Tumeurs du sein triple-négatives , Humains , Tumeurs du sein triple-négatives/anatomopathologie , Tumeurs du sein triple-négatives/métabolisme , Pyrroles/pharmacologie , Femelle , Lignée cellulaire tumorale , Dioxoles/pharmacologie , Cellules souches tumorales/métabolisme , Cellules souches tumorales/effets des médicaments et des substances chimiques , Cellules souches tumorales/anatomopathologie , Transition épithélio-mésenchymateuse/effets des médicaments et des substances chimiques , Fongicides industriels/pharmacologie , Fongicides industriels/toxicité , Mouvement cellulaire/effets des médicaments et des substances chimiques , Métastase tumorale , Cellules géantes/effets des médicaments et des substances chimiques , Cellules géantes/métabolisme , Cellules géantes/anatomopathologie , Résistance aux médicaments antinéoplasiques/effets des médicaments et des substances chimiques , Protéine p53 suppresseur de tumeur/métabolisme , Protéine p53 suppresseur de tumeur/génétique , Antinéoplasiques/pharmacologie , Survie cellulaire/effets des médicaments et des substances chimiques , Prolifération cellulaire/effets des médicaments et des substances chimiques , Cycle cellulaire/effets des médicaments et des substances chimiquesRÉSUMÉ
USP7 is one of the most studied deubiquitinating enzymes, which is involved in the regulation of multiple cell signaling pathways and has been shown to be associated with the occurrence and progression of a variety of cancers. Inhibitors targeting USP7 have been studied by several teams, but most of them lack selectivity and have low activities. Herein, we reported a serious of pyrrole[2,3-d]pyrimidin-4-one derivatives through scaffold hopping of recently reported 4-hydroxypiperidine compounds. The representative compound Z33 (YCH3124) exhibited highly potent USP7 inhibition activity as well as anti-proliferative activity against four kinds of cancer cell lines. Further study revealed that YCH3124 effectively inhibited the downstream USP7 pathway and resulted in the accumulation of both p53 and p21 in a dose-dependent manner. Notably, YCH3124 disrupted cell cycle progression through restricting G1 phase and induced significant apoptosis in CHP-212 cells. In summary, our efforts provided a series of novel pyrrole[2,3-d]pyrimidin-4-one analogs as potent USP7 inhibitors with excellent anti-cancer activity.
Sujet(s)
Antinéoplasiques , Prolifération cellulaire , Tests de criblage d'agents antitumoraux , Pyrimidines , Pyrroles , Ubiquitin-specific peptidase 7 , Humains , Ubiquitin-specific peptidase 7/antagonistes et inhibiteurs , Ubiquitin-specific peptidase 7/métabolisme , Antinéoplasiques/pharmacologie , Antinéoplasiques/composition chimique , Antinéoplasiques/synthèse chimique , Pyrroles/pharmacologie , Pyrroles/composition chimique , Pyrroles/synthèse chimique , Prolifération cellulaire/effets des médicaments et des substances chimiques , Relation structure-activité , Pyrimidines/pharmacologie , Pyrimidines/composition chimique , Pyrimidines/synthèse chimique , Lignée cellulaire tumorale , Structure moléculaire , Relation dose-effet des médicaments , Apoptose/effets des médicaments et des substances chimiques , Découverte de médicament , Pyrimidinones/pharmacologie , Pyrimidinones/composition chimique , Pyrimidinones/synthèse chimique , Cycle cellulaire/effets des médicaments et des substances chimiquesRÉSUMÉ
Although substantial advances have been obtained in the pharmacological treatment of cystic fibrosis (CF) with the approval of Kaftrio, a combination of two correctors (VX-661, VX-445) and one potentiator (VX-770), new modulators are still needed to rescue F508del and other CFTR mutants with trafficking defects. We have previously identified PP compounds based on a tricyclic core as correctors with high efficacy in the rescue of F508del-CFTR on native epithelial cells of CF patients, particularly in combination with class 1 correctors (VX-809, VX-661). Compound PP028 was found as a lead candidate for the high rescue of F508del-CFTR and used for mechanistic insight indicating that PP028 behaves as a class 3 corrector, similarly to VX-445. From the exploration of the chemical space around the hit structure, based on iterative cycles of chemical synthesis and functional testing, the class of 6,9-dihydro-5H-pyrrolo [3,2-h]quinazolines with corrector activity was discovered. Within a series of 38 analogues, two derivatives emerged as promising candidates and used for further insight to assess the mechanism of action. Both compounds, decorated with a benzensulfonylamino group at the pyrimidine moiety, were able to generate a dose-dependent increase in CFTR function, particularly in the presence of VX-809. Half-effective concentrations (EC50) were in the single digit micromolar range and decreased in the presence of VX-809 thus indicating a synergistic interaction with class 1 correctors. Synergy was also observed with corr-4a (class 2 corrector) but not with VX-445 and PP028 (class 3 correctors) indicating that the new compounds behave as class 3 correctors. These results suggest that tricyclic pyrrolo-quinazolines interact with CFTR at a site different from that of VX-809 and represent a novel class of CFTR correctors suitable for combinatorial pharmacological treatments for the basic defect in CF.
Sujet(s)
Protéine CFTR , Mucoviscidose , Quinazolines , Mucoviscidose/traitement médicamenteux , Mucoviscidose/métabolisme , Protéine CFTR/métabolisme , Protéine CFTR/génétique , Humains , Quinazolines/pharmacologie , Quinazolines/composition chimique , Quinazolines/synthèse chimique , Relation structure-activité , Relation dose-effet des médicaments , Structure moléculaire , Pyrroles/pharmacologie , Pyrroles/composition chimique , Pyrroles/synthèse chimique , MutationRÉSUMÉ
Pseudomonas aeruginosa has already been stipulated as a "critical" pathogen, emphasizing the urgent need for researching and developing novel antibacterial agents due to multidrug resistance. Bacterial biofilm formation facilitates cystic fibrosis development and restricts the antibacterial potential of many current antibiotics. The capacity of P. aeruginosa to form biofilms and resist antibiotics is closely correlated with quorum sensing (QS). Bacterial QS is being contemplated as a promising target for developing novel antibacterial agents. QS inhibitors are a promising strategy for treating chronic infections. This study reported that the active compound PT22 (1H-pyrrole-2,5-dicarboxylic acid) isolated from Perenniporia tephropora FF2, one endophytic fungus from Areca catechu L., presents QS inhibitory activity against P. aeruginosa. Combined with gentamycin or piperacillin, PT22 functions as a novel antibiotic accelerant against P. aeruginosa. PT22 (0.50 mg/mL, 0.75 mg/mL, and 1.00 mg/mL) reduces the production of QS-related virulence factors, such as pyocyanin and rhamnolipid, and inhibits biofilm formation of P. aeruginosa PAO1 instead of affecting its growth. The architectural disruption of the biofilms was confirmed by visualization through scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Real-time quantitative PCR (RT-qPCR) indicated that PT22 significantly attenuated the expression of QS-related genes followed by docking analysis of molecules against QS activator proteins. PT22 dramatically increased the survival rate of Galleria mellonella. PT22 combined with gentamycin or piperacillin presents significant inhibition of biofilm formation and eradication of mature biofilm compared to monotherapy, which was also confirmed by visualization through SEM and CLSM. After being treated with PT22 combined with gentamycin or piperacillin, the survival rates of G. mellonella were significantly increased compared to those of monotherapy. PT22 significantly enhanced the susceptibility of gentamycin and piperacillin against P. aeruginosa PAO1. Our results suggest that PT22 from P. tephropora FF2 as a potent QS inhibitor is a candidate antibiotic accelerant to combat the antibiotic resistance of P. aeruginosa.
Sujet(s)
Antibactériens , Biofilms , Pseudomonas aeruginosa , Pyrroles , Détection du quorum , Pseudomonas aeruginosa/effets des médicaments et des substances chimiques , Détection du quorum/effets des médicaments et des substances chimiques , Antibactériens/pharmacologie , Biofilms/effets des médicaments et des substances chimiques , Pyrroles/pharmacologie , Animaux , Facteurs de virulence/génétique , Endophytes/composition chimique , Endophytes/métabolisme , Tests de sensibilité microbienne , Diacides carboxyliques/pharmacologie , Simulation de docking moléculaire , Pyocyanine/métabolismeRÉSUMÉ
Plant diseases caused by pathogenic fungi seriously affect the yield and quality of crops, cause huge economic losses, and pose a considerable threat to global food security. Phenylpyrrole analogues were designed and synthesized based on alkaloid lycogalic acid. All target compounds were characterized by 1H NMR, 13C NMR, and HRMS. Their antifungal activities against seven kinds of phytopathogenic fungi were evaluated. The results revealed that most compounds had broad-spectrum fungicidal activities at 50 µg/mL; 14 compounds displayed more than 60% fungicidal activities against Rhizoctonia cerealis and Sclerotinia sclerotiorum, and in particular, the fungicidal activities of compounds 8g and 8h against Rhizoctonia cerealis were more than 90%, which could be further developed as lead agents for water-soluble fungicides. The molecular docking results indicate that compounds 8g and 8h can interact with 14α-demethylase (RcCYP51) through hydrogen bonding with strong affinity.
Sujet(s)
Alcaloïdes , Antifongiques , Conception de médicament , Simulation de docking moléculaire , Pyrroles , Rhizoctonia , Antifongiques/pharmacologie , Antifongiques/synthèse chimique , Antifongiques/composition chimique , Pyrroles/composition chimique , Pyrroles/pharmacologie , Pyrroles/synthèse chimique , Alcaloïdes/composition chimique , Alcaloïdes/pharmacologie , Alcaloïdes/synthèse chimique , Rhizoctonia/effets des médicaments et des substances chimiques , Relation structure-activité , Tests de sensibilité microbienne , Structure moléculaire , Ascomycota/effets des médicaments et des substances chimiquesRÉSUMÉ
Tumor-associated macrophages (TAMs) and cancer-associated fibroblasts (CAFs) are known to play supportive roles in tumor development and progression, but their interactions in colorectal cancer (CRC) remain unclear. Here, we investigated the effects of colon-cancer-derived CAFs on TAM differentiation, migration, and tumor immunity, both in vitro and in vivo. When co-cultured with monocytes, CAFs attracted monocytes and induced their differentiation into M2 macrophages. Immunohistology of surgically resected human CRC specimens and orthotopically transplanted mouse tumors revealed a correlation between numbers of CAFs and numbers of M2 macrophages. In a mouse model of CRC orthotopic transplantation, treatment with an inhibitor of the colony-stimulating factor-1 receptor (PLX3397) depleted M2 macrophages and increased CD8-positive T cells infiltrating the tumor nest. While this treatment had a minor effect on tumor growth, combining PLX3397 with anti-PD-1 antibody significantly reduced tumor growth. RNA-seq following combination therapy showed activation of tumor immunity. In summary, CAFs are involved in the induction and mobilization of M2 macrophage differentiation in the CRC tumor immune microenvironment, and the combination of cancer immunotherapy and PLX3397 may represent a novel therapeutic option for CRC.
Sujet(s)
Fibroblastes associés au cancer , Différenciation cellulaire , Tumeurs colorectales , Inhibiteurs de points de contrôle immunitaires , Macrophages , Microenvironnement tumoral , Animaux , Tumeurs colorectales/traitement médicamenteux , Tumeurs colorectales/immunologie , Tumeurs colorectales/anatomopathologie , Tumeurs colorectales/métabolisme , Fibroblastes associés au cancer/métabolisme , Fibroblastes associés au cancer/effets des médicaments et des substances chimiques , Fibroblastes associés au cancer/immunologie , Souris , Inhibiteurs de points de contrôle immunitaires/pharmacologie , Inhibiteurs de points de contrôle immunitaires/usage thérapeutique , Différenciation cellulaire/effets des médicaments et des substances chimiques , Humains , Macrophages/immunologie , Macrophages/métabolisme , Macrophages/effets des médicaments et des substances chimiques , Microenvironnement tumoral/effets des médicaments et des substances chimiques , Microenvironnement tumoral/immunologie , Aminopyridines/pharmacologie , Aminopyridines/usage thérapeutique , Pyrrolidines/pharmacologie , Pyrrolidines/usage thérapeutique , Lignée cellulaire tumorale , Macrophages associés aux tumeurs/immunologie , Macrophages associés aux tumeurs/effets des médicaments et des substances chimiques , Macrophages associés aux tumeurs/métabolisme , Modèles animaux de maladie humaine , Pyrroles/pharmacologie , Pyrroles/usage thérapeutique , Femelle , Lymphocytes T CD8+/immunologie , Lymphocytes T CD8+/métabolisme , Lymphocytes T CD8+/effets des médicaments et des substances chimiquesRÉSUMÉ
Angiogenesis is critical for rheumatoid arthritis (RA) progression. The effects of tofacitinib, a JAK-STAT inhibitor used for RA treatment, on angiogenesis in RA are unclear. We, therefore, evaluated the levels of angiogenic factors in two systems of a human co-culture of fibroblast (HT1080) and monocytic (U937) cell lines treated with tofacitinib and in serum samples from RA patients before and after six months of tofacitinib treatment. Tofacitinib reduced CD147 levels, matrix metalloproteinase-9 (MMP-9) activity, and angiogenic potential but increased endostatin levels and secreted proteasome 20S activity. In vitro, tofacitinib did not change CD147 mRNA but increased miR-146a-5p expression and reduced STAT3 phosphorylation. We recently showed that CD147 regulates the ability of MMP-9 and secreted proteasome 20S to cleave collagen XVIIIA into endostatin. We show here that tofacitinib-enhanced endostatin levels are mediated by CD147, as CD147-siRNA or an anti-CD147 antibody blocked proteasome 20S activity. The correlation between CD147 and different disease severity scores supported this role. Lastly, tofacitinib reduced endostatin' s degradation by inhibiting cathepsin S activity and recombinant cathepsin S reversed this in both systems. Thus, tofacitinib inhibits angiogenesis by reducing pro-angiogenic factors and enhancing the anti-angiogenic factor endostatin in a dual effect mediated partly through CD147 and partly through cathepsin S.
Sujet(s)
Polyarthrite rhumatoïde , Antigènes CD147 , Cathepsines , Endostatines , Pipéridines , Pyrimidines , Humains , Antigènes CD147/métabolisme , Antigènes CD147/génétique , Pipéridines/pharmacologie , Endostatines/métabolisme , Endostatines/pharmacologie , Pyrimidines/pharmacologie , Cathepsines/métabolisme , Polyarthrite rhumatoïde/traitement médicamenteux , Polyarthrite rhumatoïde/métabolisme , Polyarthrite rhumatoïde/anatomopathologie , Matrix metalloproteinase 9/métabolisme , Matrix metalloproteinase 9/génétique , Facteur de transcription STAT-3/métabolisme , Néovascularisation pathologique/métabolisme , Néovascularisation pathologique/traitement médicamenteux , Inhibiteurs de l'angiogenèse/pharmacologie , Femelle , Adulte d'âge moyen , Mâle , Pyrroles/pharmacologie , Lignée cellulaireRÉSUMÉ
Hematopoietic stem cell transplantation can deliver therapeutic proteins to the central nervous system (CNS) through transplant-derived microglia-like cells. However, current conditioning approaches result in low and slow engraftment of transplanted cells in the CNS. Here we optimized a brain conditioning regimen that leads to rapid, robust, and persistent microglia replacement without adverse effects on neurobehavior or hematopoiesis. This regimen combines busulfan myeloablation and six days of Colony-stimulating factor 1 receptor inhibitor PLX3397. Single-cell analyses revealed unappreciated heterogeneity of microglia-like cells with most cells expressing genes characteristic of homeostatic microglia, brain-border-associated macrophages, and unique markers. Cytokine analysis in the CNS showed transient inductions of myeloproliferative and chemoattractant cytokines that help repopulate the microglia niche. Bone marrow transplant of progranulin-deficient mice conditioned with busulfan and PLX3397 restored progranulin in the brain and eyes and normalized brain lipofuscin storage, proteostasis, and lipid metabolism. This study advances our understanding of CNS repopulation by hematopoietic-derived cells and demonstrates its therapeutic potential for treating progranulin-dependent neurodegeneration.
Sujet(s)
Busulfan , Microglie , Progranulines , Animaux , Microglie/métabolisme , Microglie/effets des médicaments et des substances chimiques , Progranulines/métabolisme , Progranulines/génétique , Souris , Busulfan/pharmacologie , Transplantation de cellules souches hématopoïétiques , Aminopyridines/pharmacologie , Encéphale/métabolisme , Pyrroles/pharmacologie , Souris de lignée C57BL , Cellules souches hématopoïétiques/métabolisme , Cellules souches hématopoïétiques/effets des médicaments et des substances chimiques , Cellules souches hématopoïétiques/cytologie , Transplantation de moelle osseuse , Mâle , Système nerveux central/métabolisme , Souris knockout , Conditionnement pour greffe/méthodes , Analyse sur cellule unique , Cytokines/métabolisme , Récepteur de facteur de croissance granulocyte-macrophage/métabolisme , Récepteur de facteur de croissance granulocyte-macrophage/génétique , Récepteur de facteur de croissance granulocyte-macrophage/antagonistes et inhibiteursRÉSUMÉ
The cannabinoid receptor type 1 (CB1R) is pivotal within the endocannabinoid system regulating various signaling cascades with effects in appetite regulation, pain perception, memory formation, and thermoregulation. Still, understanding of CB1R's cellular signaling, distribution, and expression dynamics is very fragmentary. Real-time visualization of CB1R is crucial for addressing these questions. Selective drug-like CB1R ligands with a defined pharmacological profile were investigated for the construction of CB1R fluorescent probes using a reverse design-approach. A modular design concept with a diethyl glycine-based building block as the centerpiece allowed for the straightforward synthesis of novel probe candidates. Validated by computational docking studies, radioligand binding, and cAMP assay, this systematic approach allowed for the identification of novel pyrrole-based CB1R fluorescent probes. Application in fluorescence-based target-engagement studies and live cell imaging exemplify the great versatility of the tailored CB1R probes for investigating CB1R localization, trafficking, pharmacology, and its pathological implications.
Sujet(s)
Colorants fluorescents , Récepteur cannabinoïde de type CB1 , Colorants fluorescents/composition chimique , Colorants fluorescents/synthèse chimique , Récepteur cannabinoïde de type CB1/métabolisme , Humains , Simulation de docking moléculaire , Cellules HEK293 , Ligands , Pyrroles/composition chimique , Pyrroles/pharmacologie , Pyrroles/synthèse chimique , Relation structure-activité , AMP cyclique/métabolismeRÉSUMÉ
Brown rot, caused by Monilinia species, is a destructive disease of pome and stone fruits that can lead to significant losses in production. Disease management is mainly based on fungicide applications during the growing season. Fludioxonil, a "new-generation reduced-risk fungicide", is one of the most important fungicide used. The objectives of the present study were to compare and determine the toxicity of fludioxonil to selected M. laxa, M. fructigena and M. fructicola isolates, to test its effectiveness in detached fruits and to assess its effectiveness under practical control conditions. A total of 27 isolates (10 isolates of M. laxa, 8 of M. fructigena and 9 of M. fructicola) were tested for sensitivity to fludioxonil in vitro. Isolates from each species exhibited a homogeneous response to the fungicide, while differences among the different species were determined. Based on calculated resistance factors (RF), the examined isolates were classified into two categories: sensitive and moderately resistant. In vivo testing of the effectiveness of the label concentration of fludioxonil on detached fruit did not reveal differences between isolates classified into different sensitivity categories; fludioxonil used at the label concentration (0.1%) inhibited decay development 93.5 to 100%, regardless of the isolate category. Field trials revealed the very high efficacy of fludioxonil in preventing brown rot on fruits, ranging from 92.2 to 100 for peach, 90.7 to 97.3 for plum and 84.9 to 91.9% for sour cherry. In conclusion, fludioxonil was highly effective according to in vitro sensitivity tests and when used under practical field conditions for brown rot control.
Sujet(s)
Ascomycota , Dioxoles , Fongicides industriels , Maladies des plantes , Pyrroles , Fongicides industriels/pharmacologie , Dioxoles/pharmacologie , Pyrroles/pharmacologie , Maladies des plantes/microbiologie , Maladies des plantes/prévention et contrôle , Ascomycota/effets des médicaments et des substances chimiques , Fruit/microbiologie , Résistance des champignons aux médicamentsRÉSUMÉ
With an objective to improve the profiles of the 1st generation non-basic MCHR1 antagonists, a lean design approach of replacing the bicyclic thienopyrimidine core with a monocyclic pyrrol-2-one chemotype was examined in the context of reducing aromatic ring count, while also contemplating enhanced flexibility as a means of decreasing flat character. The new compounds exhibited potent antagonism up to the sub-nanomolar range, thereby implying that the monocyclic ring could effectively serve as an effective bioisostere of the bicyclic system. The prototype compound 2m offered benefits like improved potency, reduced half-life, and enhanced solubility, while also demonstrating >5% reduction in weight gain in rats, thereby providing proof-of-concept for this new class of compounds as anti-obesity agents.