scholarly journals Discovery of Novel Potential Reversible Peptidyl Arginine Deiminase Inhibitor

2019 ◽  
Vol 20 (9) ◽  
pp. 2174 ◽  
Author(s):  
Ardita Aliko ◽  
Marta Kamińska ◽  
Katherine Falkowski ◽  
Ewa Bielecka ◽  
Malgorzata Benedyk-Machaczka ◽  
...  
Keyword(s):  
Posttranslational Modification ◽  
Treatment Strategies ◽  
Arginine Deiminase ◽  
In Vitro Assays ◽  
Promising Direction ◽  
Potential Inhibitors ◽  
Pad Inhibitor ◽  
Inhibition Potency ◽  
Micromolar Range

Citrullination, a posttranslational modification, is catalyzed by peptidylarginine deiminases (PADs), a unique family of enzymes that converts peptidyl-arginine to peptidyl-citrulline. Overexpression and/or increased PAD activity is observed in rheumatoid arthritis (RA), Alzheimer’s disease, multiple sclerosis, and cancer. Moreover, bacterial PADs, such as Porphyromonas gingivalis PAD (PPAD), may have a role in the pathogenesis of RA, indicating PADs as promising therapeutic targets. Herein, six novel compounds were examined as potential inhibitors of human PAD4 and PPAD, and compared to an irreversible PAD inhibitor, Cl-amidine. Four of the tested compounds (compounds 2, 3, 4, and 6) exhibited a micromolar-range inhibition potency against PAD4 and no effect against PPAD in the in vitro assays. Compound 4 was able to inhibit the PAD4-induced citrullination of H3 histone with higher efficiency than Cl-amidine. In conclusion, compound 4 was highly effective and presents a promising direction in the search for novel RA treatment strategies.

scholarly journals Small-Molecule Inhibitors of the Rce1p CaaX Protease

2007 ◽  
Vol 12 (7) ◽  
pp. 983-993 ◽  
Author(s):  
Surya P. Manandhar ◽  
Emily R. Hildebrandt ◽  
Walter K. Schmidt
Keyword(s):  
Small Molecule ◽  
Small Molecule Inhibitors ◽  
Proteolytic Processing ◽  
In Vivo Studies ◽  
In Vitro Assays ◽  
Selective Agents ◽  
Wild Type Yeast ◽  
Micromolar Range

The Rce1p protease is required for the maturation of the Ras GTPase and certain other isoprenylated proteins and is considered a chemotherapeutic target. To identify new small-molecule inhibitors of Rce1p, the authors screened the National Cancer Institute Diversity Set compound library using in vitro assays to monitor the proteolytic processing of peptides derived from Ras and the yeast a-factor mating pheromone. Of 46 inhibitors initially identified with a Ras-based assay, only 9 were effective in the pheromone-based assay. The IC50 values of these 9 compounds were in the low micromolar range for both yeast (6-35 µM) and human Rce1p (0.4-46 µM). Four compounds were somewhat Rce1p selective in that they partially inhibited the Ste24p protease and did not inhibit Ste14p isoprenylcysteine carboxyl methyltransferase, 2 enzymes also involved in the maturation of isoprenylated proteins. The remaining 5 compounds inhibited all 3 enzymes. The 2 most Rce1p-selective agents were ineffective trypsin inhibitors, further supporting the specificity of these agents for Rce1p. The 5 least specific compounds formed colloidal aggregates, a proposed common feature of promiscuous inhibitors. Interestingly, the most specific Rce1p inhibitor also formed a colloidal aggregate. In vivo studies revealed that treatment of wild-type yeast with 1 compound induced a Ras2p delocalization phenotype that mimics observed effects in rce1 ste24 null yeast. The 9 compounds identified in this study represent new tools for understanding the enzymology of postisoprenylation-modifying enzymes and provide new insight for the future development of Rce1p inhibitors. ( Journal of Biomolecular Screening 2007:983-993)

scholarly journals Considerations and Technical Pitfalls in the employment of the MTT Assay to Evaluate Photosensitizers for Photodynamic Therapy

2021 ◽  
Vol 11 (6) ◽  
pp. 2603
Author(s):  
Edith Alejandra Carreño ◽  
Anael Viana Pinto Alberto ◽  
Cristina Alves Magalhães de Souza ◽  
Heber Lopes de Mello ◽  
Andrea Henriques-Pons ◽  
...  
Keyword(s):  
Photodynamic Therapy ◽  
Mtt Assay ◽  
Neuroblastoma Cell ◽  
Treatment Strategies ◽  
Chemical Substance ◽  
In Vitro Assays ◽  
Diphenyltetrazolium Bromide ◽  
Number Of Publications ◽  
Neuroblastoma Cell Lines

Photodynamic therapy (PDT) combines light, a photosensitizing chemical substance, and molecular oxygen to elicit cell death and is employed in the treatment of a variety of diseases, including cancer. The development of PDT treatment strategies requires in vitro assays to develop new photosensitizers. One such assay is the MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide developed in 1983 and widely used in PDT studies. Despite the exponential growth in the number of publications, a uniform MTT protocol for use in the PDT area is lacking. Herein, we list and standardize the conditions to evaluate the photosensitizer methylene blue (MB) in glioblastoma and neuroblastoma cell lines. In addition, we review technical pitfalls and identify several variables that must be taken into consideration in order to provide accurate results with MTT. We conclude that for each cell line we must have a dose-response curve using the MTT assay and good controls for the standardization. Additionally, the optimal values of the time and cell density must be in the linear range of the curve to avoid errors. We describe all relevant points and outline the best normalization techniques to observe the differences between treatments.

scholarly journals Peptidylarginine Deiminase inhibition abolishes the production of large extracellular vesicles fromGiardia intestinalis, affecting host-pathogen interactions by hindering adhesion to host cells

2019 ◽  
Author(s):  
Bruno Gavinho ◽  
Izadora Volpato Rossi ◽  
Ingrid Evans-Osses ◽  
Sigrun Lange ◽  
Marcel Ivan Ramirez
Keyword(s):  
Extracellular Vesicles ◽  
Mammalian Cells ◽  
Deep Understanding ◽  
Arginine Deiminase ◽  
Host Cells ◽  
Peptidylarginine Deiminase ◽  
Host Pathogen Interactions ◽  
Host Pathogen ◽  
Pad Inhibitor

AbstractGiardia intestinalisis an anaerobic protozoan that is an important etiologic agent of inflammation-driven diarrhea worldwide. Although self-limiting, a deep understanding of the factors involved in the pathogenicity that produces the disruption of the intestinal barrier remains unknown. There is evidence that under diverse conditions, the parasite is capable of shedding extracellular vesicles (EVs) which could modulate the physiopathology of giardiasis. Here we describe new insights ofG. intestinalisEV production, revealing its capacity to shed two different enriched EV populations (large and small extracellular vesicles) and identified a relevant adhesion function associated only with the larger population. Our work also aimed at assessing the influences of two recently identified inhibitors of EV release in mammalian cells, namely peptidylarginine deiminase (PAD) inhibitor and cannabidiol (CBD), on EV release fromGiardiaand their putative effects on host-pathogen interactions. PAD-inhibitor Cl-amidine and CBD were both able to effectively reduce EV shedding, the PAD-inhibitor specifically affecting the release of large extracellular vesicles and interfering within vitrohost-pathogen interactions. The strong efficacy of the PAD-inhibitor onGiardiaEV release indicates a phylogenetically conserved pathway of PAD-mediated EV release, most likely affecting theGiardiaarginine deiminase (GiADI) homolog of mammalian PADs. While there is still much to learn aboutG. intestinalisinteraction with its host, our results suggest that large and small EVs may be differently involved in protozoa communication, and that EV-inhibitor treatment may be a novel strategy for recurrent giardiasis treatment.

Design, synthesis and evaluation of potential inhibitors for poly(ADP-ribose) polymerase members 1 and 14

2020 ◽  
Vol 12 (24) ◽  
pp. 2179-2190
Author(s):  
Caleb M Kam ◽  
Amanda L Tauber ◽  
Stephan M Levonis ◽  
Stephanie S Schweiker
Keyword(s):  
Inhibitory Activity ◽  
In Silico ◽  
Parp Inhibitors ◽  
In Vitro Assays ◽  
Inhibiting Activity ◽  
Design Synthesis ◽  
In Silico Studies ◽  
Amine Compounds ◽  
Potential Inhibitors

Poly(ADP-ribose) polymerase (PARP) members PARP1 and PARP14 belong to an 18-member superfamily of post-translational modifying enzymes. A library of 9 novel non-NAD analog amine compounds was designed, synthesized and evaluated for inhibitory activity against PARP1 and PARP14. Both in silico studies and in vitro assays identified compound 2 as a potential PARP1 inhibitor, inhibiting activity by 93 ± 2% (PARP14 inhibition: 0 ± 6%), and 7 as a potential PARP14 inhibitor, inhibiting activity by 91 ± 2% (PARP1 inhibition: 18 ± 4%), at 10-μm concentration. Key in silico interactions with TYR907 in PARP1 and TYR1620 and TYR1646 in PARP14 have been identified. Compound 2 and compound 7 have been identified as potential leads for the development of selective PARP inhibitors.

Docking Simulations Exhibit Bortezomib and other Boron-containing Peptidomimetics as Potential Inhibitors of SARS-CoV-2 Main Protease

2020 ◽  
Vol 14 ◽  
Author(s):  
Iván R Vega-Valdez ◽  
Melvin N Rosalez ◽  
José M Santiago-Quintana ◽  
Eunice D Farfán-García ◽  
Marvin A Soriano-Ursúa
Keyword(s):  
Crystal Structure ◽  
In Vitro Assays ◽  
Docking Simulations ◽  
Chemical Agents ◽  
Main Protease ◽  
Drug Designing ◽  
Potential Inhibitors ◽  
Key Residues

Background:: Treatment of the COVID19 pandemic requires drug development. Boron-containing compounds are attractive chemical agents, some of them act as proteases inhibitors. Objective:: The present study explores the role of boronic moieties in molecules interacting on the binding site of the SARS-CoV-2 main protease. Methods:: Conventional docking procedure was applied by assaying boron-free and boron-containing compounds on the recently reported crystal structure of SARS-CoV-2 main protease (PDB code: 6LU7). The set of 150 ligands includes bortezomib and inhibitors of coronavirus proteases. Results:: Most of the tested compounds share contact with key residues and poses on the cleavage pocket. Those compounds with a boron atom in its structure were often estimated with higher affinity than boron-free analogues. Conclusion:: Interactions and the affinity of boron-containing peptidomimetics strongly suggest boron-moieties increases affinity on the main protease, as it should be tested by in vitro assays. A Bis-boron-containing compound previously tested as active on SARS-virus protease and bortezomib were identified as potent ligands. These advances may be relevant for drug designing, in addition as to the suggestion of testing available boron-containing drugs in patients with COVID19 infection.

In silico and in vitro assays reveal potential inhibitors against 3CLpro main protease of SARS-CoV-2

2021 ◽  
pp. 1-12
Author(s):  
Eldhose Iype ◽  
Jisha Pillai U ◽  
Indresh Kumar ◽  
Silvia V. Gaastra-Nedea ◽  
Ramachandran Subramanian ◽  
...  
Keyword(s):  
In Silico ◽  
In Vitro Assays ◽  
Main Protease ◽  
Potential Inhibitors

scholarly journals Selected by Bioinformatics and Molecular Docking Analysis, Dhea and 2-14,15-Eg are Effective Against Cholangiocarcinoma

2021 ◽  
Author(s):  
Lei Qin ◽  
Jun Kuai ◽  
Fang Yang ◽  
Lu Yang ◽  
Peisheng Sun ◽  
...  
Keyword(s):  
Gene Expression ◽  
Molecular Docking ◽  
Therapeutic Effect ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
In Vitro Assays ◽  
Ppi Network ◽  
Network Analyses ◽  
Potential Inhibitors

Abstract Object To identify novel targets for the diagnosis, treatment and prognosis of cholangiocarcinoma, we screen ideal lead compounds and preclinical drug candidates with MYC inhibitory effect from the ZINC database, and verify the therapeutic effect of Dhea and 2-14,15-Eg on cholangiocarcinoma.Methods The gene expression profiles of GSE132305, GSE89749, and GSE45001 were obtained respectively from the Gene Expression Omnibus database. The DEGs were identified by comparing the gene expression profiles of cholangiocarcinoma and normal tissues. GO, KEGG analysis and PPI network analyses were performed. LibDock, ADME and toxicity prediction, molecular docking and molecular dynamics simulations were used to identify potential inhibitors of MYC. Moreover, in vitro, MTT assay, colony-forming assay and the scratch assay were performed to verify the therapeutic effect of Dhea and 2-14,15-Eg.Results PPI network analysis showed that ALB, MYC, APOB, IGF1 and KNG1 were hub genes, of which MYC was mainly studied in this study. A battery of computer-aided virtual techniques showed that Dhea and 2-14,15-Eg have lower rodent carcinogenicity, Ames mutagenicity, developmental toxicity potential, and high tolerance to cytochrome P4502D6, as well as could exist stably in natural circumstances. In vitro assays showed that Dhea and 2-14,15-Eg inhibited cholangiocarcinoma cellular viability, proliferation, and migration via inducing cholangiocarcinoma cells apoptosis.Conclusion This study suggested that Dhea and 2-14,15-Eg were novel potential inhibitors of MYC targeting, as well as are a promising drug in dealing with cholangiocarcinoma and have a perspective application.

scholarly journals The Effect of Metadherin on NF-κB Activation and Downstream Genes in Ovarian Cancer

2020 ◽  
Vol 29 ◽  
pp. 096368972090550 ◽  
Author(s):  
Chunhong Rong ◽  
Yanfen Shi ◽  
Jun Huang ◽  
Xinyue Wang ◽  
Risa Shimizu ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Gynecological Cancer ◽  
Treatment Strategies ◽  
In Vitro Assays ◽  
Nf Kappa B ◽  
Tnf Α ◽  
Novel Therapeutic Strategies ◽  
Tumor Types

Ovarian cancer (OC) is the most aggressive gynecological cancer. Even with the advances in detection and therapeutics, it still remains clinically challenging and there is a pressing need to identify novel therapeutic strategies. In searching for rational molecular targets, we identified metadherin (MTDH), a multifunctional gene associated with several tumor types but previously unrecognized in OC. In this study, we found the MTDH is overexpressed in OC tissues. Through in vitro assays with overexpression cells, we characterized the role of MTDH. We confirmed MTDH stable overexpression significantly increased the expression of TNF-α, IL-6, IL-8, IL-10, and IL-1β. Interestingly, NF-kappa-B (NF-κB) and MTDH were found in a feed-forward loop motif. Thus, our findings support the notion that the MTDH and NF-κB signaling network contributes to OC traits. MTDH represents a new OC-associated gene that can contribute to insights of OC biology and suggests other treatment strategies.

Neurofilaments and Paired Helical Filaments

1985 ◽  
Vol 43 ◽  
pp. 740-743
Author(s):  
J. Metuzals
Keyword(s):  
Animal Model ◽  
Posttranslational Modifications ◽  
Posttranslational Modification ◽  
Giant Axon ◽  
Paired Helical Filaments ◽  
Squid Giant Axon ◽  
In Vitro Experiments ◽  
Thin Sections ◽  
Structural Relationships

It has been demonstrated that the neurofibrillary tangles in biopsies of Alzheimer patients, composed of typical paired helical filaments (PHF), consist also of typical neurofilaments (NF) and 15nm wide filaments. Close structural relationships, and even continuity between NF and PHF, have been observed. In this paper, such relationships are investigated from the standpoint that the PHF are formed through posttranslational modifications of NF. To investigate the validity of the posttranslational modification hypothesis of PHF formation, we have identified in thin sections from frontal lobe biopsies of Alzheimer patients all existing conformations of NF and PHF and ordered these conformations in a hypothetical sequence. However, only experiments with animal model preparations will prove or disprove the validity of the interpretations of static structural observations made on patients. For this purpose, the results of in vitro experiments with the squid giant axon preparations are compared with those obtained from human patients. This approach is essential in discovering etiological factors of Alzheimer's disease and its early diagnosis.

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