scholarly journals Identification of potential TNF-α inhibitors: from in silico to in vitro studies

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Komal Zia ◽  
Sajda Ashraf ◽  
Almas Jabeen ◽  
Maria Saeed ◽  
Mohammad Nur-e-Alam ◽  
...  
Keyword(s):  
Small Molecule ◽  
Therapeutic Potential ◽  
Pharmacophore Model ◽  
Docking Studies ◽  
Immune Functions ◽  
Binding Interaction ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
Potential Inhibitors

AbstractTumor Necrosis Factor Alpha (TNF-α) is a pleiotropic pro-inflammatory cytokine. It act as central biological regulator in critical immune functions, but its dysregulation has been linked with a number of diseases. Inhibition of TNF-α has considerable therapeutic potential for diseases such as cancer, diabetes, and especially autoimmune diseases. Despite the fact that many small molecule inhibitors have been identified against TNF-α, no orally active drug has been reported yet which demand an urgent need of a small molecule drug against TNF-α. This study focuses on the development of ligand-based selective pharmacophore model to perform virtual screening of plant origin natural product database for the identification of potential inhibitors against TNF-α. The resultant hits, identified as actives were evaluated by molecular docking studies to get insight into their potential binding interaction with the target protein. Based on pharmacophore matching, interacting residues, docking score, more affinity towards TNF-α with diverse scaffolds five compounds were selected for in vitro activity study. Experimental validation led to the identification of three chemically diverse potential compounds with the IC50 32.5 ± 4.5 µM, 6.5 ± 0.8 µM and 27.4 ± 1.7 µM, respectively.

scholarly journals Thalidomide-based inhibitor for TNF-α: designing and Insilico evaluation

2022 ◽  
Vol 8 (1) ◽  
Author(s):  
Biradar Shivaleela ◽  
S. C. Srushti ◽  
S. J. Shreedevi ◽  
R. L. Babu
Keyword(s):  
Small Molecule ◽  
Small Molecule Inhibitors ◽  
Inflammatory Diseases ◽  
Binding Capacity ◽  
Preliminary Investigation ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
Monocytes And Macrophages ◽  
Potential Ligand

Abstract Background Inflammatory diseases are the vast array of disorders caused by inflammation. During most inflammatory events, many cytokines expressions were modulated, and one such cytokine is tumor necrosis factor-alpha (TNF-α). TNF-α is mainly secreted by monocytes and macrophages. Notably, it has been proposed as a therapeutic target for several diseases. The anti-TNF biology approach is mainly based on monoclonal antibodies. The fusion protein and biosimilars are prevalent in treating inflammation for decades. Only a few small molecule inhibitors are available to inhibit the expression of TNF-α, and one such promising drug was thalidomide. Therefore, the study was carried out to design thalidomide-based small molecule inhibitors for TNF-α. The main objective of our study is to design thalidomide analogs to inhibit TNF-α using the insilico approach. Results Several thalidomide analogs were designed using chemsketch. After filtration of compounds through ‘Lipinski rule of 5’ by Molinspiration tool, as a result, five compounds were selected. All these compounds were subjected to molecular docking, and the study showed that all five compounds had good binding energy. However, based on ADMET predictions, two compounds (S3 and S5) were eliminated. Conclusions Our preliminary results suggest that S1, S2, S4 compounds showed potential ligand binding capacity with TNF-α and, interestingly, with limited or no toxicity. Our preliminary investigation and obtained results have fashioned more interest for further in vitro studies.

scholarly journals Targeting dysregulation signatures of p53-MDM2 interactions to identify newer small molecule inhibitors for cancer therapy: Insight from computational direction

2021 ◽  
Author(s):  
Prosper Obed Chukwuemeka ◽  
Haruna Isiyaku Umar ◽  
Opeyemi Iwaloye ◽  
Oluwaseyi Matthew Oretade ◽  
Christopher Busayo Olowosoke ◽  
...  
Keyword(s):  
Binding Affinity ◽  
Small Molecule ◽  
Small Molecule Inhibitors ◽  
Therapeutic Option ◽  
Pharmacophore Model ◽  
In Vivo Studies ◽  
Hydrogen Bond Acceptor ◽  
Potential Inhibitors

Abstract Dysregulation of the p53-MDM2 interactions has been implicated in majority of human tumors presenting a target for finding small molecule inhibitors. In this study, a training set of 17 experimentally tested inhibitors of MDM2 was used to develop series of pharmacophore models among which a four-featured (AHRR_1) model with one hydrogen bond acceptor, one hydrophobic group and two aromatic ring features and characterized by a survival score of 4.176 was considered significant among the top ranked generated hypothesis. Further, the model was validated by an external set of actives and decoy molecules and was found to exhibit encouraging statistical attributes (such as AUC > 0.7, BEDROC > 0.5 and EF > 1.0 etc). The model was used to screen the ZINC compound database, from the database, the top best 1375 hits satisfying the pharmacophore model was were docked to MDM2 protein to identify the likely interactions of the compounds as well as their binding affinity with MDM2. Further, druglikeness and pharmacokinetic properties screening on top-ranked compounds with higher binding affinity than reference inhibitors revealed four compounds (ZINC02639178, ZINC38933175, ZINC77969611, and ZINC06752762) with suitable pharmacological properties including low ligand toxicity. Investigation of the dynamic behaviour of each candidate inhibitors in complex with MDM2 via molecular dynamic simulation suggested ZINC02639178 and ZINC06752762 as the most potential inhibitors. Thus, these compounds may emerged as therapeutic option for cancer treatment after extensive in vitro and in vivo studies.

Pharmacophore Modeling and Docking Studies to Investigate Potential Leads for the Development of β -Secretase APP Cleavage Enzyme-1 (BACE-1) Inhibitors

2019 ◽  
Vol 16 (7) ◽  
pp. 775-784
Author(s):  
Richa Arya ◽  
Satya Prakash Gupta ◽  
Sarvesh Paliwal ◽  
Swapnil Sharma ◽  
Kirtika Madan ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Medical Condition ◽  
Pharmacophore Model ◽  
Pharmacophore Modeling ◽  
Docking Studies ◽  
Pyridine Derivative ◽  
Cleavage Enzyme ◽  
Bace 1

Background: Alzheimer’s disease is a medical condition with detrimental brain health. It is majorly diagnosed in aging individuals plaque in β) characterized by accumulated Amyloidal beta (A 1 BACE) 1 secretase APP cleavage enzyme βneurological areas. The ) is the target of choice that can be exploited to find drugs against Alzheimer’s disease. Methods: A series of BACE-1 inhibitors with reported binding constant were considered for the development of a feature based pharmacophore model. Results: The good correlation coefficient (r=0.91) and RMSD of 0.93 was observed with 30 compounds in training set. The model was validated internally (r2test=0.76) as well as externally by Fischer validation. The pharmacophore based virtual screening retrieved compounds that were docked and biologically evaluated. Conclusion: The three structurally diverse molecules were tested by in-vitro method. The pyridine derivative with highest fit value (6.9) exhibited IC50 value of 2.70 µM and thus was found to be the most promising lead molecule as BACE-1 inhibitor.

scholarly journals In Vitro Evaluation of the Anti-Inflammatory Effect of KMUP-1 and in Vivo Analysis of Its Therapeutic Potential in Osteoarthritis

Biomedicines ◽  
2021 ◽  
Vol 9 (6) ◽  
pp. 615
Author(s):  
Shang-En Huang ◽  
Erna Sulistyowati ◽  
Yu-Ying Chao ◽  
Bin-Nan Wu ◽  
Zen-Kong Dai ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Inflammatory Responses ◽  
Therapeutic Potential ◽  
Antioxidant Properties ◽  
Serum Levels ◽  
Gene Expressions ◽  
Tnf Α ◽  
Anti Inflammatory

Osteoarthritis is a degenerative arthropathy that is mainly characterized by dysregulation of inflammatory responses. KMUP-1, a derived chemical synthetic of xanthine, has been shown to have anti-inflammatory and antioxidant properties. Here, we aimed to investigate the in vitro anti-inflammatory and in vivo anti-osteoarthritis effects of KMUP-1. Protein and gene expressions of inflammation markers were determined by ELISA, Western blotting and microarray, respectively. RAW264.7 mouse macrophages were cultured and pretreated with KMUP-1 (1, 5, 10 μM). The productions of TNF-α, IL-6, MMP-2 and MMP- 9 were reduced by KMUP-1 pretreatment in LPS-induced inflammation of RAW264.7 cells. The expressions of iNOS, TNF-α, COX-2, MMP-2 and MMP-9 were also inhibited by KMUP-1 pretreatment. The gene expression levels of TNF and COX families were also downregulated. In addition, KMUP-1 suppressed the activations of ERK, JNK and p38 as well as phosphorylation of IκBα/NF-κB signaling pathways. Furthermore, SIRT1 inhibitor attenuated the inhibitory effect of KMUP-1 in LPS-induced NF-κB activation. In vivo study showed that KMUP-1 reduced mechanical hyperalgesia in monoiodoacetic acid (MIA)-induced rats OA. Additionally, KMUP-1 pretreatment reduced the serum levels of TNF-α and IL-6 in MIA-injected rats. Moreover, macroscopic and histological observation showed that KMUP-1 reduced articular cartilage erosion in rats. Our results demonstrated that KMUP-1 inhibited the inflammatory responses and restored SIRT1 in vitro, alleviated joint-related pain and cartilage destruction in vivo. Taken together, KMUP-1 has the potential to improve MIA-induced articular cartilage degradation by inhibiting the levels and expression of inflammatory mediators suggesting that KMUP-1 might be a potential therapeutic agent for OA.

scholarly journals N-Acetylcysteine (NAC): Impacts on Human Health

Antioxidants ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 967
Author(s):  
Micaely Cristina dos Santos Tenório ◽  
Nayara Gomes Graciliano ◽  
Fabiana Andréa Moura ◽  
Alane Cabral Menezes de Oliveira ◽  
Marília Oliveira Fonseca Goulart
Keyword(s):  
Human Health ◽  
Therapeutic Potential ◽  
Experimental Studies ◽  
Primary Role ◽  
Necrosis Factor Alpha ◽  
Biochemical Basis ◽  
Tnf Α ◽  
Factor Alpha ◽  
Anti Inflammatory

N-acetylcysteine (NAC) is a medicine widely used to treat paracetamol overdose and as a mucolytic compound. It has a well-established safety profile, and its toxicity is uncommon and dependent on the route of administration and high dosages. Its remarkable antioxidant and anti-inflammatory capacity is the biochemical basis used to treat several diseases related to oxidative stress and inflammation. The primary role of NAC as an antioxidant stems from its ability to increase the intracellular concentration of glutathione (GSH), which is the most crucial biothiol responsible for cellular redox imbalance. As an anti-inflammatory compound, NAC can reduce levels of tumor necrosis factor-alpha (TNF-α) and interleukins (IL-6 and IL-1β) by suppressing the activity of nuclear factor kappa B (NF-κB). Despite NAC’s relevant therapeutic potential, in several experimental studies, its effectiveness in clinical trials, addressing different pathological conditions, is still limited. Thus, the purpose of this chapter is to provide an overview of the medicinal effects and applications of NAC to human health based on current therapeutic evidence.

scholarly journals Interaction of Antimicrobial Peptide Temporin L with Lipopolysaccharide In Vitro and in Experimental Rat Models of Septic Shock Caused by Gram-Negative Bacteria

2006 ◽  
Vol 50 (7) ◽  
pp. 2478-2486 ◽  
Author(s):  
Andrea Giacometti ◽  
Oscar Cirioni ◽  
Roberto Ghiselli ◽  
Federico Mocchegiani ◽  
Fiorenza Orlando ◽  
...  
Keyword(s):  
Septic Shock ◽  
Antimicrobial Peptide ◽  
Gram Negative Bacteria ◽  
Amphibian Skin ◽  
Necrosis Factor Alpha ◽  
Gram Negative ◽  
Rat Models ◽  
E Coli ◽  
Tnf Α

ABSTRACT Sepsis remains a major cause of morbidity and mortality in hospitalized patients, despite intense efforts to improve survival. The primary lead for septic shock results from activation of host effector cells by endotoxin, the lipopolysaccharide (LPS) associated with cell membranes of gram-negative bacteria. For these reasons, the quest for compounds with antiendotoxin properties is actively pursued. We investigated the efficacy of the amphibian skin antimicrobial peptide temporin L in binding Escherichia coli LPS in vitro and counteracting its effects in vivo. Temporin L strongly bound to purified E. coli LPS and lipid A in vitro, as proven by fluorescent displacement assay, and readily penetrated into E. coli LPS monolayers. Furthermore, the killing activity of temporin L against E. coli was progressively inhibited by increasing concentrations of LPS added to the medium, further confirming the peptide's affinity for endotoxin. Antimicrobial assays showed that temporin L interacted synergistically with the clinically used β-lactam antibiotics piperacillin and imipenem. Therefore, we characterized the activity of temporin L when combined with imipenem and piperacillin in the prevention of lethality in two rat models of septic shock, measuring bacterial growth in blood and intra-abdominal fluid, endotoxin and tumor necrosis factor alpha (TNF-α) concentrations in plasma, and lethality. With respect to controls and single-drug treatments, the simultaneous administration of temporin L and β-lactams produced the highest antimicrobial activities and the strongest reduction in plasma endotoxin and TNF-α levels, resulting in the highest survival rates.

scholarly journals Suppression of tumor necrosis factor-alpha-induced neutrophil adherence responses by essential oils

2003 ◽  
Vol 12 (6) ◽  
pp. 323-328 ◽  
Author(s):  
Shigeru Abe ◽  
Naho Maruyama ◽  
Kazumi Hayama ◽  
Hiroko Ishibashi ◽  
Shigeharu Inoue ◽  
...  
Keyword(s):  
Essential Oils ◽  
Tumor Necrosis ◽  
Neutrophil Activation ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
Neutrophil Adherence ◽  
Factor Alpha ◽  
Anti Inflammatory ◽  
Necrosis Factor

Background:In aromatherapy, essential oils are used as anti-inflammatory remedies, but experimental studies on their action mechanisms are very limited.Aims:To assess their anti-inflammatory activities, effects of essential oils on neutrophil activation were examinedin vitro.Methods:Neutrophil activation was measured by tumor necrosis factor-alpha (TNF-α)-induced adherence reaction of human peripheral neutrophils.Results:All essential oils tested at 0.1% concentration suppressed TNF-α-induced neutrophil adherence, and, in particular, lemongrass, geranium and spearmint oils clearly lowered the reaction even at 0.0125%. Similar inhibitory activities for the neutrophil adherence were obtained by their major constituent terpenoids: citral, geraniol, citronellol and carvone. In contrast, very popular essential oils, tea tree oil and lavender oil, did not display the inhibitory activity at the concentration.Conclusion:Thus, some essential oils used as anti-inflammatory remedies suppress neutrophil activation by TNF-α at a low concentration (0.0125-0.025%)in vitro.

scholarly journals Ανακάλυψη αναστολέων για την πρωτεΐνη TNF-α σαν ενώσεις οδηγούς για φάρμακα έναντι της ρευματοειδoύς αρθρίτιδας

2017 ◽  
Author(s):  
Ανθούλα Μέττου
Keyword(s):  
Tumour Necrosis ◽  
Docking Studies ◽  
Structure Property ◽  
E Coli ◽  
Tnf Α ◽  
Structure Property Relationship ◽  
Relationship Model ◽  
Dissociation Constant Kd ◽  
Necrosis Factor

Αντικείμενο της παρούσας διατριβής είναι η ανακάλυψη νέων μικρομοριακών αναστολέων για τον Παράγοντα Νέκρωσης Όγκων (Tumour Necrosis Factor, TNF). Ο TNF έχει συσχετισθεί με αυτοάνοσες διαταραχές όπως ψωρίαση, νόσος του Crohn, νόσος του Alzheimer και παίζει καθοριστικό ρόλο στην παθογένεια της ρευματοειδούς αρθρίτιδας. Η αντι-TNF θεραπεία χρησιμοποιείται για ασθένειες όπως η αγκυλοποιητική σπονδυλαρθρίτιδα, η ρευματοειδής και ψωριασική αρθρίτιδα, η ψωρίαση, η φλεγμονώδης νόσος του εντέρου κ.ά. Το SPD304 είναι ένα μικρό μόριο που αναστέλλει τη δράση του TNF καθώς αλληλεπιδρά με το τριμερές της πρωτεΐνης, προωθεί τον αποτριμερισμό της και έτσι προκύπτει ένα ανενεργό διμερές. Το SPD304 είναι τοξικό και για τον λόγο αυτό δεν είναι δυνατό να χρησιμοποιηθεί σαν φαρμακευτική ουσία στην αντι-TNF θεραπεία αλλά σαν βάση για την ανακάλυψη νέων μικρομοριακών αναστολέων που εμποδίζουν τον τριμερισμό της πρωτεΐνης. Επομένως, η ανάπτυξη μικρομοριακών αναστολέων του TNF χαμηλής τοξικότητας παραμένει ένας διακαής στόχος.Το αντικείμενο του προγράμματος RAlead ήταν η ανακάλυψη μικρομοριακών αναστολέων του TNF και σε αυτό το πλαίσιο σχεδιάστηκε και συντέθηκε μια σειρά αναλόγων του SPD304 με σκοπό τη μείωση της τοξικότητας και την ενίσχυση της βιοδραστικότητας. Επιπρόσθετα, μελετήθηκαν εμπορικά διαθέσιμες ενώσεις που προέκυψαν από in silico μελέτες ελλιμενισμού (docking studies) αλλά και ενώσεις που σχεδιάστηκαν βάσει των ενδιάμεσων αποτελεσμάτων των μελετών πρόσδεσης. Για την αξιολόγηση των πιθανών προσδετών μετρήθηκε η σταθερά διάσπασης (dissociation constant, Kd) με φασματοσκοπία φθορισμού. Παράλληλα, μελετήθηκε ο μηχανισμός πρόσδεσης με την τεχνική της Ισόθερμης Θερμιδομετρίας Τιτλοδότησης (ITC). Επίσης μελετήθηκε κρυσταλλογραφικά ο τρόπος πρόσδεσης του SPD304 στην πρωτεΐνη TNF που χρησιμοποιήθηκε και στις μελέτες πρόσδεσης. Για όλες τις παραπάνω μελέτες προηγήθηκε έκφραση του TNF σε κύτταρα E-coli και καθαρισμός βάσει ενός πρωτόκολλου που αναπτύχθηκε.Ένα σοβαρό πρόβλημα που εμφανίστηκε και αντιμετωπίστηκε με επιτυχία κατά τη διάρκεια αυτής της εργασίας, ήταν η χαμηλή υδατική διαλυτότητα των υπό μελέτη ουσιών. Μελετήθηκαν συγκεκριμένες τεχνικές για την ενίσχυση της διαλυτότητας και δημιουργήθηκε ένα πρωτόκολλο διάλυσης μικρών μορίων. Το πρωτόκολλο αυτό διαμορφώθηκε έτσι ώστε να είναι συμβατό με τις βιοχημικές δοκιμές πρόσδεσης όπως και με την πρωτεΐνη. Εκτός από τις in vitro βιοχημικές δοκιμές οι τεχνικές αυτές μπορούν να αποτελέσουν ένα δυνατό εργαλείο και στην κρυστάλλωση πρωτεϊνικών συμπλόκων με μικρά φαρμακευτικά μόρια. Μελετήθηκε μια σειρά 86 αναλόγων του SPD304 καθώς και μία σειρά 60 πιθανών αναστολέων που προέκυψαν από μελέτες ελλιμενισμού. Η πλειοψηφία των μορίων αυτών δεν ήταν αρκετά υδατοδιαλυτή έτσι ώστε να μετρηθεί η σταθερά διάσπασης. Με τις μελέτες διαλυτότητας κατέστη δυνατό οι περισσότερες από αυτές να διαλυθούν αρκετά έτσι ώστε να είναι δυνατή η μέτρηση της σταθεράς διάσπασης. Στο πλαίσιο των μελετών διαλυτότητας αξιολογήθηκε η επίδραση της προσθήκης συνδιαλυτών (DMSO, PEG3350) και άλλων προσθέτων (β-κυκλοδεξτρίνης) όπου φάνηκε ότι η PEG3350 είναι πιο αποτελεσματική από το DMSO για μετρίως αδιάλυτα μόρια και ότι η β-κυκλοδεξτρίνη είναι καταλληλότερη σε περιπτώσεις εξαιρετικά αδιάλυτων μορίων. Επίσης μελετήθηκε η επιρροή του χρόνου διάλυσης, της αρχικής φάσης των χημικών ενώσεων (στερεή/ υγρή) και του φιλτραρίσματος των δειγμάτων πριν από τη μέτρηση της διαλυτότητας. Οι μελέτες δραστικότητας ανέδειξαν περίπου 30 ενώσεις με δραστικότητα καλύτερη εκείνης του SPD304 (<5μM) και 20 ενώσεις με εφάμιλλη του SPD304 δραστικότητα (~5μM). Μερικές από αυτές είχαν σαφώς μειωμένη τοξικότητα. Το ποσοστό των ενώσεων που ήταν αδιάλυτες σε καθαρά υδατικά διαλύματα το οποίο είχε υπολογιστεί σε 76%, βάσει των μελετών διαλυτότητας, με τη χρήση συνδιαλυτών και άλλων τεχνικών ενίσχυσης της διαλυτότητας μειώθηκε σε 20%. Αυτό σημαίνει ότι έγινε εφικτή η ικανοποιητική διάλυση του 80% των ενώσεων έτσι ώστε να είναι δυνατή η μέτρηση της σταθεράς διάσπασής τους. Με βάση τα διαθέσιμα δεδομένα διαλυτότητας δημιουργήθηκε ένα μοντέλο πρόβλεψης διαλυτοτήτων (Novamechanics) το οποίο στηρίζεται στην ταξινόμηση διαχωρίζοντας τις αδιάλυτες από τις διαλυτές ενώσεις. Το μοντέλο Ποσοτικής Συσχέτισης Δομής Ιδιότητας (predictive Quantitative Structure Property Relationship model) στοχεύει στην πρόβλεψη της διαλυτότητας πιθανών μικρομοριακών αναστολέων του TNF σε υδατικά διαλύματα παρουσία 5% DMSO. Η απλότητα της προτεινόμενης μεθόδου την καθιστά ευρέως εφαρμόσιμη σε υπολογιστική διαλογή (virtual screening) και εξαγωγή δεδομένων (data mining) για την ταυτοποίηση διαλυτών μορίων.

Abstract 319: Small Molecule Gβγ Inhibition Attenuates Cardiac Fibroblast Inflammatory and Pro-Fibrotic Signaling

2014 ◽  
Vol 115 (suppl_1) ◽  
Author(s):  
Joshua G Travers ◽  
Fadia A Kamal ◽  
Michael S Burhans ◽  
Burns C Blaxall
Keyword(s):  
Myocardial Fibrosis ◽  
Inflammatory Cytokines ◽  
Small Molecule ◽  
Therapeutic Potential ◽  
Interstitial Fibrosis ◽  
Pressure Overload ◽  
Cardiac Fibroblast ◽  
Ventricular Compliance ◽  
Pro Inflammatory Cytokines

Heart failure (HF) is a devastating disease characterized by chamber remodeling, interstitial fibrosis and reduced ventricular compliance. Prolonged sympathetic overstimulation promotes excess signaling through G-protein Gβγ subunits and ultimately results in pathologic GRK2-mediated β-adrenergic receptor (β-AR) downregulation. We have recently demonstrated the therapeutic potential of the small molecule Gβγ-GRK2 inhibitor Gallein in limiting HF progression. Pathologic activation of the cardiac fibroblast (CF) induces the transition to a myofibroblast phenotype, which plays a fundamental role in myocardial fibrosis and remodeling. We hypothesized that Gβγ-GRK2 inhibition plays an important functional role in the CF to attenuate pathologic CF activation, inflammation and interstitial fibrosis. To explore the effect of Gβγ-GRK2 inhibition on inflammation and pro-fibrotic signaling, mice were subjected to 7 days of transverse aortic constriction, a pressure-overload model of HF. In addition to the attenuation in overall cardiac hypertrophy, animals treated with Gallein displayed reduced expression of pro-inflammatory cytokines, including macrophage inflammatory protein 1 alpha (MIP-1α) and MIP-1β, along with Interleukin-6, as assessed by qPCR. Gallein-treated animals also exhibited diminished pro-fibrotic signaling, as evidenced by a reduction in the expression of TGFβ, a major driver of myocardial fibrosis, and decreased expression of collagen. To recapitulate these findings in vitro, primary adult mouse ventricular fibroblasts were pathologically stimulated using Isoproterenol (ISO, β-AR agonist) or Angiotensin II and treated +/- Gallein for 24 hours. CFs treated with Gallein displayed an analogous reduction in the expression of these pro-inflammatory cytokines and collagen. In summary, these data suggest a potential therapeutic role for small molecule Gβγ-GRK2 inhibition in limiting pathologic myofibroblast activation, inflammation and interstitial fibrosis. We believe this fibroblast-targeted approach will lead to the refinement of existing targets and compounds, and possibly the generation of novel therapeutics for the treatment of HF.

scholarly journals Pharmacological Inhibition of Caspase-8 Suppresses Inflammation-Induced Angiogenesis in the Cornea

Biomolecules ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 210
Author(s):  
Yunzhe Tian ◽  
He Li ◽  
Xiuxing Liu ◽  
Lihui Xie ◽  
Zhaohao Huang ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Therapeutic Potential ◽  
Corneal Neovascularization ◽  
Caspase 8 ◽  
Inhibitory Effects ◽  
Toll Like Receptor 4 ◽  
Pharmacological Inhibition ◽  
Cytokines And Chemokines ◽  
Tnf Α

Inflammation-induced angiogenesis is closely related to many diseases and has been regarded as a therapeutic target. Caspase-8 has attracted increasing attention for its immune properties and therapeutic potential in inflammatory disorders. The aim of our study is to investigate the clinical application of pharmacological inhibition of caspase-8 and the underlying molecular mechanisms in inflammation-induced angiogenesis in the cornea. A model of alkali burn (AB)-induced corneal neovascularization (CNV) in C57BL/6 wild-type (WT) mice and toll-like receptor 4 knockout (Tlr4-/-) mice was used. We found that AB increased caspase-8 activity and the pharmacological inhibition of caspase-8 exerted substantial inhibitory effects on CNV, with consistent decreases in caspase-8 activity, inflammatory cell infiltration, macrophage recruitment and activation, VEGF-A, TNF-α, IL-1β, MIP-1, and MCP-1 expression in the cornea. In vitro, caspase-8 mediated TLR4–dependent chemokines and VEGF-A production by macrophages. The TLR4 knockout significantly alleviated CNV, suppressed caspase-8 activity and downregulated expression of inflammatory cytokines and chemokines after AB. Taken together, these findings provide the first demonstration that the pharmacological inhibition of caspase-8 suppresses inflammation-induced angiogenesis and support the use of a pharmacological caspase-8 inhibitor as a novel clinical treatment for CNV and other angiogenic disorders.

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