scholarly journals Targeting Kinases in Fasciola hepatica: Anthelminthic Effects and Tissue Distribution of Selected Kinase Inhibitors

2020 ◽  
Vol 7 ◽  
Author(s):  
Carolin M. Morawietz ◽  
Hicham Houhou ◽  
Oliver Puckelwaldt ◽  
Laura Hehr ◽  
Domenic Dreisbach ◽  
...  
Keyword(s):  
Tissue Distribution ◽  
Protein Kinases ◽  
Egg Production ◽  
Kinase Inhibitors ◽  
Fasciola Hepatica ◽  
Human Cancer ◽  
Oral Route ◽  
New Drugs ◽  
Uptake Kinetic

Protein kinases have been discussed as promising druggable targets in various parasitic helminths. New drugs are also needed for control of fascioliasis, a food-borne trematode infection and worldwide spread zoonosis, caused by the liver fluke Fasciola hepatica and related species. In this study, we intended to move protein kinases more into the spotlight of Fasciola drug research and characterized the fasciolicidal activity of two small-molecule inhibitors from human cancer research: the Abelson tyrosine kinase (ABL-TK) inhibitor imatinib and the polo-like 1 (PLK1) inhibitor BI2536. BI2536 reduced viability of 4-week-old immature flukes in vitro, while adult worms showed a blockade of egg production. Together with a significantly higher transcriptional expression of PLK1 in adult compared to immature worms, this argues for a role of PLK1 in fluke reproduction. Both fluke stages expressed ABL1-TK transcripts at similar high levels and were affected by imatinib. To study the uptake kinetic and tissue distribution of imatinib in F. hepatica, we applied matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) for the first time in this parasite. Drug imaging revealed the accumulation of imatinib in different fluke tissues from 20 min to 12 h of exposure. Furthermore, we show that imatinib is metabolized to N-desmethyl imatinib by F. hepatica, a bioactive metabolite also found in humans. Besides the vitellarium, gastrodermal tissue showed strong signal intensities. In situ hybridization demonstrated the gastrodermal presence of abl1 transcripts. Finally, we assessed transcriptional changes of physiologically important genes in imatinib-treated flukes. Moderately increased transcript levels of a gene encoding a multidrug resistance protein were detected, which may reflect an attempt to defend against imatinib. Increased expression levels of the cell cycle dependently expressed histone h2b and of two genes encoding superoxide dismutases (SODs) were also observed. In summary, our pilot study demonstrated cross-stage activity of imatinib but not BI2536 against immature and adult F. hepatica in vitro; a fast incorporation of imatinib within minutes, probably via the oral route; and imatinib-induced expression changes of physiologically relevant genes. We conclude that kinases are worth analyzing in more detail to evaluate the potential as therapeutic targets in F. hepatica.

scholarly journals Kinase Inhibitors of Novel Pyridopyrimidinone Candidates: Synthesis and In Vitro Anticancer Properties

2019 ◽  
Vol 2019 ◽  
pp. 1-10 ◽  
Author(s):  
Nagy M. Khalifa ◽  
Mohamed A. Al-Omar ◽  
Hamad M. Alkahtani ◽  
Ahmed H. Bakheit
Keyword(s):  
Kinase Inhibitors ◽  
Human Cancer ◽  
Ethyl Cyanoacetate ◽  
Diethyl Malonate ◽  
Aromatic Aldehydes ◽  
Docking Studies ◽  
Braf V600e ◽  
Kinase Assay ◽  
Human Cancer Cell Lines

A new class of pyridopyrimidinone compounds containing different nitrogenous heterocycles were synthesized starting from the key precursor 2-hydrazinyl-5-phenyl-7-(pyridin-3-yl)pyrido[2,3-d]pyrimidin-4(3H)-one via condensation with series of aromatic aldehydes and cyclization using different reagents as ethyl acetoacetate, ethyl cyanoacetate, diethyl malonate, and ammonium isothiocyanate. The bioassay results showed compound 6 to be highly effective towards three human cancer cell lines (HepG2, PC-3, and HCT-116) in vitro with promising activity values (IC50: 0.5 μM) relative to the standard doxorubicin (IC50: 0.6 μM). Kinase inhibitory evaluation of compound 6 displays hopeful inhibitory action against BRAF V600E, EGFR, and PDGFRβ at100 μM. The molecular docking studies supported the initial kinase assay.

scholarly journals NITROXYNIL UNCOUPLES OXIDATIVE PHOSPHORYLATION IN THE CELL MITOCHONDRIA AND A DRUG WHEREVER INJECTABLES ARE PREFERRED OVER DRENCHES

2017 ◽  
Vol 15 (1) ◽  
pp. 45-49 ◽  
Author(s):  
M. M. Rahman ◽  
A. Kabir ◽  
S. Ahmed ◽  
M. K. Islam ◽  
M. S. Rahman ◽  
...  
Keyword(s):  
Fasciola Hepatica ◽  
Stable Solution ◽  
Production Quality ◽  
Oral Route ◽  
New Drugs ◽  
Intermediate Hosts ◽  
Management Options ◽  
Blood Sucking ◽  
Fluke Infection ◽  
The One

In the absence of effective vaccines and because of practical limitations in management options to control the snail intermediate hosts, the control of liver fluke infection and disease in livestock relies heavily on the use of flukicidal anthelmintics. Nitroxynil is a nitrate derivative from benzene compounds and presents a flukicide effect and is commonly used as the one with best spectrum from all the nitrate derivative trematocides. The drug is in fact, a low spectrum anthelmintic effective against fascioliasis, an important parasitic disease caused by flukes like Fasciola hepatica, Fasciola gigantica, affecting significantly the ruminant production by means of reducing the growth, conversion rate, milk production, quality and quantity of meat and reproduction. Nitroxynil also does hold efficacy against few roundworms mainly blood-sucking species as well as certain myiasis. It is not effective against most of the tape worms or external parasites. It is chemically constitute of yellow benzene crystals, slightly soluble in water and soluble in organic solvents, almost odorless and is characterized for being a stable solution, but it precipitates in presence of calcium and other salts. It is marketed as bright orange-red injectable solution due that by this route is six times more efficient than by oral route. Nitroxynil is a safe drug and it can be used at any stage of pregnancy. It does not affect fertility, gestation or fetus formation and in no way impede the reproductive performance of stallions. It’s presented as two salts, nitroxynil megaglumine and nitroxynil eglumine. The nitroxynil in salt is hydrosoluble having neutral pH and is exclusively administered to animals by subcutaneous route. The flukicide resistance phenomenon is already present in many countries in the same way that resistance against other anthelmintic drugs is spreading around worldwide. Because of the absence of new drugs against fluke infections, it is necessary to prove and compare the efficacy of the anthelmintics to prevent resistance development. In view of that we endeavored elucidating the latest progress of one such widely used flukicide nitroxynil.

scholarly journals MEK and ERK Kinase Inhibitors Increase Circulating Ceruloplasmin and Cause Green Serum in Rats

2016 ◽  
Vol 45 (2) ◽  
pp. 353-361 ◽  
Author(s):  
Dolores Diaz ◽  
Rama Pai ◽  
Gary Cain ◽  
Nghi La ◽  
Donna Dambach ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Kinase Inhibitors ◽  
Human Cancer ◽  
Elevated Serum ◽  
Clinical Signs ◽  
Copper Toxicity ◽  
Mitogen Activated Protein Kinase ◽  
Serum Ceruloplasmin ◽  
Mitogen Activated Protein

Inhibition of the mitogen-activated protein kinase/extracellular signal-regulated (MAPK/ERK) pathway is an attractive therapeutic approach for human cancer therapy. In the course of evaluating structurally distinct small molecule inhibitors that target mitogen-activated protein kinase kinase (MEK) and ERK kinases in this pathway, we observed an unusual, dose-related increase in the incidence of green serum in preclinical safety studies in rats. Having ruled out changes in bilirubin metabolism, we demonstrated a 2- to 3-fold increase in serum ceruloplasmin levels, likely accounting for the observed green color. This was not associated with an increase in α-2-macroglobulin, the major acute phase protein in rats, indicating that ceruloplasmin levels increased independently of an inflammatory response. Elevated serum ceruloplasmin was also not correlated with changes in total hepatic copper, adverse clinical signs, or pathology findings indicative of copper toxicity, therefore discounting copper overload as the etiology. Both ERK and MEK inhibitors led to increased ceruloplasmin secretion in rat primary hepatocyte cultures in vitro, and this increase was associated with activation of the Forkhead box, class O1 (FOXO1) transcription factor. In conclusion, increased serum ceruloplasmin induced by MEK and ERK inhibition is due to increased synthesis by hepatocytes from FOXO1 activation and results in the nonadverse development of green serum in rats.

A scanning electron microscope study on the route of entry of triclabendazole into the liver fluke,Fasciola hepatica

Parasitology ◽  
2009 ◽  
Vol 136 (5) ◽  
pp. 523-535 ◽  
Author(s):  
E. TONER ◽  
F. McCONVERY ◽  
G. P. BRENNAN ◽  
M. MEANEY ◽  
I. FAIRWEATHER
Keyword(s):  
Liver Fluke ◽  
Fasciola Hepatica ◽  
Oral Route ◽  
Drug Uptake ◽  
Scanning Electron Microscope Study ◽  
Normal Morphology ◽  
Pharmacological Studies ◽  
Marked Decline ◽  
Scanning Electron

SUMMARYStudies have been carried out to establish the relative importance of oral and trans-tegumental uptake of triclabendazole by the liver fluke,Fasciola hepatica. Experiments were designed to block either oral uptake of drug, by use of ligatures, or trans-tegumental diffusion, by allowing the drug to bind to bovine serum albumin (BSA) in the medium. Changes to the surface morphology of the tegument and gut were assessed by scanning electron microscopy. Flukes were incubatedin vitrofor 24 h in TCBZ.SO at a concentration of 15 μg/ml. Tegumental disruption in ligatured and non-ligatured flukes was similar, suggesting that closing the oral route did not affect drug uptake. The gut remained unaffected by drug treatment. When BSA (30 mg/ml) was present in the medium, there was a marked decline in the level of tegumental disruption. Again, the gut retained a normal morphology. Non-ligatured flukes were also incubated for 24 hin vitroin TCBZ.SO (15 μg/ml) in the presence of red blood cells. Oral ingestion of blood was demonstrated, although the gut surface retained a normal morphology. In contrast, the tegumental surface was severely affected by the drug. The findings support previous pharmacological studies which suggest that trans-tegumental uptake of triclabendazole predominates in the liver fluke.

In vitro maintenance of Fasciola hepatica: A factorial approach based on egg production

1969 ◽  
Vol 26 (1) ◽  
pp. 41-51 ◽  
Author(s):  
Leslie H Ractliffe ◽  
Diego Guevara-Pozo ◽  
Ramon Lopez-Roman
Keyword(s):  
Egg Production ◽  
Fasciola Hepatica ◽  
Factorial Approach ◽  
In Vitro Maintenance

scholarly journals The repositioning of epigenetic probes/inhibitors identifies new anti-schistosomal lead compounds and chemotherapeutic targets

2019 ◽  
Author(s):  
Kezia C. L. Whatley ◽  
Gilda Padalino ◽  
Helen Whiteland ◽  
Kathrin K. Geyer ◽  
Benjamin J. Hulme ◽  
...  
Keyword(s):  
Schistosoma Mansoni ◽  
Structural Genomics ◽  
Egg Production ◽  
Anthelmintic Activity ◽  
New Drugs ◽  
Next Generation ◽  
Blood Fluke ◽  
Protein Targets ◽  
Vitelline Cells

AbstractBackgroundPraziquantel represents the frontline chemotherapy used to treat schistosomiasis, a neglected tropical disease (NTD) caused by infection with macro-parasitic blood fluke schistosomes. While this drug is safe, its inability to kill all schistosome lifecycle stages within the human host often requires repeat treatments. This limitation, amongst others, has led to the search for novel anti-schistosome replacement or combinatorial chemotherapies. Here, we describe a repositioning strategy to assess the anthelmintic activity of epigenetic probes/inhibitors obtained from the Structural Genomics Consortium.Methodology/Principle findingsThirty-seven epigenetic probes/inhibitors targeting histone readers, writers and erasers were initially screened against Schistosoma mansoni schistosomula using the high-throughput Roboworm platform. At 10 µM, 14 of these 37 compounds (38%) negatively affected schistosomula motility and phenotype after 72 hours of continuous co-incubation. Subsequent dose-response titrations against schistosomula and adult worms revealed epigenetic probes targeting one reader (NVS-CECR2-1), one writer (LLY-507 and BAY-598) and one eraser (GSK-J4) to be particularly active. As LLY-507/BAY-598 (SMYD2 histone methyltransferase inhibitors) and GSK-J4 (a JMJD3 histone demethylase inhibitor) regulate an epigenetic process (protein methylation) known to be critical for schistosome development, further characterisation of these compounds/putative targets was performed. RNA interference (RNAi) of one putative LLY-507/BAY-598 S. mansoni target (Smp_000700) in adult worms replicated the compound-mediated motility and egg production defects. Furthermore, H3K36me2, a known product catalysed by SMYD2 activity, was also reduced by LLY-507 (25%), BAY-598 (23%) and siSmp_000700 (15%) treatment of adult worms. Oviposition and packaging of vitelline cells into in vitro laid eggs was also significantly affected by GSK-J4 (putative cell permeable prodrug inhibitor of Smp_034000), but not by the related structural analogue GSK-J1 (non-permeable inhibitor).Conclusion/SignificanceCollectively, these results provide further support for the development of next-generation drugs targeting schistosome epigenetic pathway components. In particular, the progression of histone methylation/demethylation modulators presents a tractable strategy for anti-schistosomal control.Author SummaryHuman schistosomiasis is caused by infection with parasitic blood fluke worms. Global control of this NTD is currently facilitated by administration of a single drug, praziquantel (PZQ). This mono-chemotherapeutic strategy of schistosomiasis control presents challenges as PZQ is not active against all human-dwelling schistosome lifecycle stages and the evolution of PZQ resistant parasites remains a theat. Therefore, new drugs to be used in combination with or in replacement of PZQ are urgently needed. Here, continuing our studies on Schistosoma mansoni epigenetic processes, we performed anthelmintic screening of 37 epigenetic probes/epigenetic inhibitors obtained from the Structural Genomics Consortium (SGC). The results of these studies highlighted that schistosome methylation/demethylation processes are acutely vulnerable. In particular, compounds affecting schistosome SMYD (LLY-507, BAY-598) or JMJD (GSK-J4) homologues are especially active on schistosomula and adult worms during in vitro phenotypic drug screens. The active epigenetic probes identified here as well as their corresponding S. mansoni protein targets offers new starting points for the development of next-generation anti-schistosomals.

scholarly journals Akt inhibition promotes autophagy and sensitizes PTEN-null tumors to lysosomotropic agents

2008 ◽  
Vol 183 (1) ◽  
pp. 101-116 ◽  
Author(s):  
Michael Degtyarev ◽  
Ann De Mazière ◽  
Christine Orr ◽  
Jie Lin ◽  
Brian B. Lee ◽  
...  
Keyword(s):  
Cell Death ◽  
Kinase Inhibitors ◽  
Human Cancer ◽  
Akt Pathway ◽  
Anticancer Efficacy ◽  
Human Cancer Cells ◽  
Cancer Cell Survival ◽  
Pathway Inhibition

Although Akt is known as a survival kinase, inhibitors of the phosphatidylinositol 3-kinase (PI3K)–Akt pathway do not always induce substantial apoptosis. We show that silencing Akt1 alone, or any combination of Akt isoforms, can suppress the growth of tumors established from phosphatase and tensin homologue–null human cancer cells. Although these findings indicate that Akt is essential for tumor maintenance, most tumors eventually rebound. Akt knockdown or inactivation with small molecule inhibitors did not induce significant apoptosis but rather markedly increased autophagy. Further treatment with the lysosomotropic agent chloroquine caused accumulation of abnormal autophagolysosomes and reactive oxygen species, leading to accelerated cell death in vitro and complete tumor remission in vivo. Cell death was also promoted when Akt inhibition was combined with the vacuolar H+–adenosine triphosphatase inhibitor bafilomycin A1 or with cathepsin inhibition. These results suggest that blocking lysosomal degradation can be detrimental to cancer cell survival when autophagy is activated, providing rationale for a new therapeutic approach to enhancing the anticancer efficacy of PI3K–Akt pathway inhibition.

JAK2 Inhibitor Therapy and Mechanisms of Response

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. SCI-36-SCI-36
Author(s):  
Ayalew Tefferi
Keyword(s):  
Clinical Trials ◽  
Kinase Inhibitors ◽  
Myeloproliferative Neoplasms ◽  
Unmet Need ◽  
New Drugs ◽  
Added Value ◽  
Current Therapy ◽  
Common Side Effect ◽  
Jak Inhibitors

Abstract Abstract SCI-36 Myeloproliferative neoplasms (MPN) are stem cell-derived clonal (or oligoclonal) hemopathies and are phenotypically characterized by abnormally increased left-shifted myeloproliferation. “BCR-ABL1-negative MPN” is an operational sub-category of MPN that includes polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF). Beginning in 2005, novel mutations involving JAK2, MPL, TET2, ASXL1, IDH1, IDH2, CBL, IKZF1, or LNK have been described in a variable proportion of patients with BCR-ABL1-negative MPN. The evidence so far suggests that none of these mutations garner the disease specificity or pathogenetic relevance otherwise displayed by BCR-ABL1. Nevertheless, considering the fact that some of these mutations exhibit high mutational frequency (e.g. JAK2 mutations) and either directly (e.g. JAK2 or MPL mutations) or indirectly (e.g. LNK or CBL mutations) induce JAK-STAT hyperactivation, it is reasonable to evaluate the therapeutic value of anti-JAK drugs in these disorders. In this regard, the ATP-mimetic small molecule kinase inhibitors have so far been the focus and those that have made it into clinical trials include TG101348, INCB018424, CEP-701, CYT387, AZD1480, SB1518, XL019 and LY2784544 (clinicaltrials.gov). Preliminary results from clinical studies suggest notable differences among these drugs in their toxicity and efficacy profiles, some of which might be linked to their variable in vitro activity against other JAK and non-JAK kinase targets and the resultant effect on JAK-STAT-dependent cytokines. For example, off-target FLT3 inhibition displayed by TG101348, CEP-701 and SB1518 might contribute to their common side effect profile of nausea, vomiting and diarrhea. Similarly, the impressive activity of INCB018424 in alleviating constitutional symptoms has been correlated with marked reduction in serum pro-inflammatory cytokines and therefore possibly related to its primarily anti-JAK1 activity whereas serum cytokine levels did not appear to be affected by either TG101348 or CEP-701. More selective anti-JAK2 drugs, such as TG101348, might be preferable in the setting of PV or ET because of their superior activity in controlling leukocytosis, thrombocytosis and JAK2V617F allele burden. Regardless, because most MPN-associated mutations do not necessarily represent the primary clonogenic event, we should curb our expectations from anti-JAK2 treatment trials and continue to pursue other treatment strategies. Furthermore, in considering the prospect of anti-JAK therapy in MPN, one should recognize the fact that life expectancy in the majority of patients with PV or ET is near-normal and disease complications are effectively managed by treatment with low-dose aspirin, phlebotomy or hydroxyurea. Therefore, controlled studies are needed to show added value and justify the risk of unknown long-term health effects associated with new drugs, including JAK inhibitors, in PV and ET. The unmet need for new treatment is in PMF where survival and quality of life are significantly worse and current therapy is inadequate. This session will i) summarize results of current anti-JAK clinical trials in MPN, ii) discuss putative mechanisms of action in terms of both therapeutic benefit and toxicity and iii) define the potential role of JAK inhibitors in the context of current therapeutic approaches for MPN. Disclosures: Off Label Use: Hydroxyurea to control myeloproliferation in myeloproliferative neoplasms Interferon-alfa to control myeloproliferation in myeloproliferative neoplasms Busulfan to control myeloproliferation Pipobroman to control myeloproliferation.

Frequent mutations in MLH1, MET, KIT, PDGFRA, and PIK3CA genes in human gastrointestinal stromal tumors.

2013 ◽  
Vol 31 (15_suppl) ◽  
pp. 10544-10544 ◽  
Author(s):  
Zhi Xu ◽  
Xinying Huo ◽  
Chuanning Tang ◽  
Hua Ye ◽  
Feng Lou ◽  
...  
Keyword(s):  
Gastrointestinal Stromal Tumors ◽  
Kinase Inhibitors ◽  
Human Cancer ◽  
New Drugs ◽  
Missense Mutations ◽  
Ion Torrent ◽  
Exon 2 ◽  
Sequencing Platform ◽  
Stromal Tumors ◽  
Frequent Mutations

10544 Background: Identifying mutations in individual tumor is critical to improve the efficacy of cancer therapy by matching targeted drugs to specific mutations. Gastrointestinal stromal tumor (GIST) is a stromal or mesenchymal subepithelial neoplasm affecting the gastrointestinal tract and it frequently contains an activating mutation in either the KIT or platelet-derived growth factor A (PDGFRA) gene. Although GIST is highly responsive to several selective tyrosine kinase inhibitors, combined use of inhibitors targeting other mutations is needed to further prolong survival. Methods: In this study, we aim to screen and identify genetic mutations in GIST for targeted therapy using the new Ion Torrent next-generation sequencing platform. Results: By utilizing the Ion Ampliseq Cancer Panel, we sequenced 737 loci from 45 cancer-related genes from DNA extracted from formalin-fixed and paraffin-embedded (FFPE) samples of 125 human gastrointestinal stromal tumors, set up stringent parameters for reliable variant calling by filtering out potential raw base calling errors, and identified frequent mutations in PDGFRA, KIT, APC, MLH1, MET, RET, and PIK3CA. Moreover, frequent missense mutations were detected in MLH1 (12%), MET (16.8%), KIT (51.2%), and PIK3CA (41.6%) genes. The mutations in MLH1 gene were frequently detected in the exon 14, while MET gene was frequently mutated in the exon 2. In addition, we identified missense mutations in other genes, including FLT3, KRAS, PDGFRA, and STK11, at lower frequencies. Conclusions: This study demonstrates the utility of using Ion Torrent sequencing to efficiently identify human cancer mutations. With the finding of the frequent mutations in MLH1 and MET genes, in addition to KIT, PDGFRA and PIK3CA, in GISTs, this study provides a molecular basis for clinically developing new drugs targeting these gene mutations for GIST therapy.

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