scholarly journals Identification of Iminium Intermediates Generation in the Metabolism of Tepotinib Using LC-MS/MS: In Silico and Practical Approaches to Bioactivation Pathway Elucidation

Molecules ◽  
2020 ◽  
Vol 25 (21) ◽  
pp. 5004 ◽  
Author(s):  
Ali S. Abdelhameed ◽  
Mohamed W. Attwa ◽  
Adnan A. Kadi
Keyword(s):  
Side Effects ◽  
Phase I ◽  
In Silico ◽  
Potassium Cyanide ◽  
Reactive Intermediates ◽  
Toxicity Assessment ◽  
Human Liver Microsome ◽  
New Drugs ◽  
Piperidine Ring

Tepotinib (Tepmetko™, Merck) is a potent inhibitor of c-Met (mesenchymal−epithelial transition factor). In March 2020, tepotinib (TEP) was approved for use in Japan for the treatment of patients who suffered from non-small cell lung cancers (NSCLC) harboring an MET exon 14 skipping alteration and have progressed after platinum-based therapy. Practical and in silico experiments were used to screen for the metabolic profile and reactive intermediates of TEP. Knowing the bioactive center and structural alerts in the TEP structure helped in making targeted modifications to improve its safety. First, the prediction of metabolism vulnerable sites and reactivity metabolic pathways was performed using the StarDrop WhichP450™ module and the online Xenosite reactivity predictor tool, respectively. Subsequently, in silico data were used as a guide for the in vitro practical work. Second, in vitro phase I metabolites of TEP were generated from human liver microsome (HLM) incubations. Testing for the generation of unstable reactive intermediates was performed using potassium cyanide as a capturing agent forming stable cyano adduct that can be characterized and identified using liquid chromatography tandem mass spectrometry (LC-MS/MS). Third, in silico toxicity assessment of TEP metabolites was performed, and structural modification was proposed to decrease their side effects and to validate the proposed bioactivation pathway using the DEREK software. Four TEP phase I metabolites and four cyano adducts were characterized. The reactive intermediate generation mechanism of TEP may provide an explanation of its adverse reactions. The piperidine ring is considered a structural alert for toxicity as proposed by the DEREK software and a Xenosite reactivity model, which was confirmed by practical experiments. Steric hindrance or isosteric replacement at α-carbon of the piperidine ring stop the bioactivation sequence that was confirmed using the DEREK software. More drug discovery studies can be performed using this perception permitting the design of new drugs with an increased safety profile. To our knowledge, this is the first study for the identification of in vitro phase I metabolites and reactive intermediates in addition to toxicological properties of the metabolites for TEP that will be helpful for the evaluation of TEP side effects and drug–drug interactions in TEP-treated patients.

scholarly journals In Silico Driven Prediction of MAPK14 Off-Targets Reveals Unrelated Proteins with High Accuracy

2020 ◽  
Author(s):  
Florian Kaiser ◽  
Maximilian G. Plach ◽  
Christoph Leberecht ◽  
Thomas Schubert ◽  
V. Joachim Haupt
Keyword(s):  
Side Effects ◽  
In Silico ◽  
Target Identification ◽  
Low Cost ◽  
Time Frame ◽  
New Drugs ◽  
Drug Candidate ◽  
Delivery Times ◽  
Target Screening

During the discovery and development of new drugs, candidates with undesired and potentially harmful side-effects can arise at all stages, which poses significant scientific and economic risks. Most of such phenotypic side-effects can be attributed to binding of the drug candidate to unintended proteins, so-called off-targets. The early identification of potential off-targets is therefore of utmost importance to mitigate any downstream risks. We showcase how the combination of knowledge-based in silico off-target screening and state-of-the-art biophysics can be applied to rapidly identify off-targets for a MAPK14 inhibitor. Out of 13 predicted off-targets, six proteins were confirmed to interact with the inhibitor in vitro, which translates to an exceptional hit rate of 46%. For two proteins, affinities in the lower micromolar range were obtained: The kinase IRE1 and the Hematopoietic Prostaglandin D Synthase, which is entirely unrelated to MAPK14 and is involved in different cell-regulatory processes. The whole off-target identification/validation pipeline can be completed as fast as within two months, excluding delivery times of proteins. These results emphasize how computational off-target screening in combination with MicroScale Thermophoresis can effectively reduce downstream development risks in a very competitive time frame and at low cost.

scholarly journals Identification of reactive intermediate formation and bioactivation pathways in Abemaciclib metabolism by LC–MS/MS: in vitro metabolic investigation

2019 ◽  
Vol 6 (1) ◽  
pp. 181714 ◽  
Author(s):  
Adnan A. Kadi ◽  
Hany W. Darwish ◽  
Hatem A. Abuelizz ◽  
Thamer A. Alsubi ◽  
Mohamed W. Attwa
Keyword(s):  
Phase I ◽  
Metabolic Profiling ◽  
Kinase Inhibitor ◽  
Liver Microsomes ◽  
Potassium Cyanide ◽  
Intermediate Formation ◽  
Reactive Intermediates ◽  
Stable Complex ◽  
Rat Liver Microsomes

Abemaciclib (Verzenio ® ) is approved as a tyrosine kinase inhibitor (TKI) for breast cancer treatment. In this study, in vitro phase I metabolic profiling of Abemaciclib (ABC) was done using rat liver microsomes (RLMs). We checked the formation of reactive intermediates in ABC metabolism using RLMs in the presence of potassium cyanide (KCN) that was used as a capturing agent for iminium reactive intermediates forming a stable complex that can be characterized by LC–MS/MS. Nine in vitro phase I metabolites and three cyano adducts were identified. The metabolic reactions involved in the formation of these metabolites and adducts are reduction, oxidation, hydroxylation and cyanide addition. The bioactivation pathway was also proposed. Knowing the electrodeficient bioactive centre in ABC structure helped in making targeted modifications to improve its safety and retain its efficacy. Blocking or isosteric replacement of α-carbon to the tertiary nitrogen atoms of piperazine ring can aid in reducing toxic side effects of ABC. No previous articles were found about in vitro metabolic profiling for ABC or structural identification of the formed reactive metabolites for ABC.

scholarly journals LC-ESI-MS/MS reveals the formation of reactive intermediates in brigatinib metabolism: elucidation of bioactivation pathways

RSC Advances ◽  
2018 ◽  
Vol 8 (3) ◽  
pp. 1182-1190 ◽  
Author(s):  
Adnan A. Kadi ◽  
Mohamed W. Attwa ◽  
Hany W. Darwish
Keyword(s):  
Phase I ◽  
Reactive Intermediates ◽  
Piperidine Ring ◽  
Esi Ms

Four phase I BGB metabolites and three cyano adducts for BGB were detected using LC-MS/MS. The piperidine ring was found to be responsible for BGB bioactivation and the bioactivation pathways are proposed.

scholarly journals Phase I Clinical Trial Results of Auranofin, a Novel Antiparasitic Agent

2016 ◽  
Vol 61 (1) ◽  
Author(s):  
Edmund V. Capparelli ◽  
Robin Bricker-Ford ◽  
M. John Rogers ◽  
James H. McKerrow ◽  
Sharon L. Reed
Keyword(s):  
Rheumatoid Arthritis ◽  
Phase I ◽  
Phase I Trial ◽  
New Drugs ◽  
Term Therapy ◽  
Parasitic Infections ◽  
Blood Levels ◽  
Content Type

ABSTRACT Under an NIH priority to identify new drugs to treat class B parasitic agents, we performed high-throughput screens, which identified the activity of auranofin (Ridaura) against Entamoeba histolytica and Giardia intestinalis, major causes of water- and foodborne outbreaks. Auranofin, an orally administered, gold (Au)-containing compound that was approved by the FDA in 1985 for treatment of rheumatoid arthritis, was effective in vitro and in vivo against E. histolytica and both metronidazole-sensitive and -resistant strains of Giardia. We now report the results of an NIH-sponsored phase I trial to characterize the pharmacokinetics (PK) and safety of auranofin in healthy volunteers using modern techniques to measure gold levels. Subjects received orally 6 mg (p.o.) of auranofin daily, the recommended dose for rheumatoid arthritis, for 7 days and were followed for 126 days. Treatment-associated adverse events were reported by 47% of the subjects, but all were mild and resolved without treatment. The mean gold maximum concentration in plasma (C max) at day 7 was 0.312 μg/ml and the half-life (t 1/2) 35 days, so steady-state blood levels would not be reached in short-term therapy. The highest concentration of gold, 13 μM (auranofin equivalent), or more than 25× the 50% inhibitory concentration (IC50) for E. histolytica and 4× that for Giardia, was in feces at 7 days. Modeling of higher doses (9 and 21 mg/day) was performed for systemic parasitic infections, and plasma gold levels of 0.4 to 1.0 μg/ml were reached after 14 days of treatment at 21 mg/day. This phase I trial supports the idea of the safety of auranofin and provides important PK data to support its potential use as a broad-spectrum antiparasitic drug. (This study has been registered at ClinicalTrials.gov under identifier NCT02089048.)

scholarly journals FBDD: In-silico STRATEGY TO INHIBIT MPRO ACTIVITY USING DRUGS FROM PREVIOUS OUTBREAKS

2021 ◽  
Vol 9 (4) ◽  
pp. 472-480
Author(s):  
Gauravi N Trivedi ◽  
◽  
Janhavi T Karlekar ◽  
Khushbu Dhimmar ◽  
Hetal kumar Panchal ◽  
...  
Keyword(s):  
Drug Development ◽  
In Silico ◽  
Drug Targets ◽  
Transmission Rate ◽  
Respiratory Illness ◽  
Primary Treatment ◽  
Rational Drug Design ◽  
New Drugs ◽  
Main Protease

Main protease (Mpro) and Spike (S) proteins are said potential drug targets of COVID-19. Pneumonia like respiratory illness caused by SARS-CoV-2 is spreading rapidly due to its replication and transmission rate. Protease is the protein that is involved in both replication and transcription. Since CoV-2 shares, genomic similarity with CoV and MERS-CoV, drugs from previous outbreaks are used as primary treatment of the disease. In-silico drug development strategies are said to be faster and effective than in-vitro with a lesser amount of risk factors. Fragment Based Drug Designing (FBDD), also known as rational drug design in which a potential target protein is selected and docked with a lead-like molecule that eventually leads to drug development. Nine (9) drugs that are currently being used to treat patients of coronavirus were selected in this study from the latest literature review and fragmented as per rules followed by crosslinking of drug fragments using editor tools. These native drugs and synthesized drugs were then docked against the main protease. Results of the study revealed that one of the crosslinked lead-like compounds showed a higher binding affinity (∆G) more than any of the native compounds. Further, the results of this study suggested that the combination of potential drugs can be an effective way to develop new drugs to treat a deadly disease.

scholarly journals Thinking Out of the Box: On the Ability of Zea mays L. to Biotrasform Aflatoxin B1 Into Its Modified Forms

2021 ◽  
Vol 11 ◽  
Author(s):  
Laura Righetti ◽  
Enrico Rolli ◽  
Luca Dellafiora ◽  
Gianni Galaverna ◽  
Michele Suman ◽  
...  
Keyword(s):  
Zea Mays ◽  
Phase I ◽  
Aflatoxin B1 ◽  
In Silico ◽  
Zea Mays L ◽  
Limited Information ◽  
Maize Plants ◽  
Modified Forms

While aflatoxin metabolism in animals has been clarified, very limited information is so far available on the possible biotransformation occurring in plants. Therefore, this work aimed at investigating whether AFB1 metabolites could occur in field-grown infected maize and the putative role of Zea mays L. metabolism in their production. For such scope, asymptomatic in vitro–grown plantlets and in silico evaluations of plant transforming enzymes were used to pinpoint how plants may handle these compounds. Our data demonstrated the role of maize plants in the production of Phase I hydroxylated aflatoxins, including, among others, AFM1, AFM2, and aflatoxicol, and suggest that plant cytochromes may be involved in this biotransformation of AFB1.

scholarly journals The Ethyl Acetate Extraction Obtained from Podocarpus Nagikernel Meal with Anticancer Activity

2021 ◽  
Vol 14 (1) ◽  
pp. 363-366
Author(s):  
Yuchen Xiao ◽  
Jianping Yong ◽  
Yang Yang ◽  
Canzhong Lu
Keyword(s):  
Side Effects ◽  
Anticancer Activity ◽  
Anticancer Drugs ◽  
Biological Activities ◽  
Wide Spectrum ◽  
Public Health Problem ◽  
New Drugs ◽  
Major Public Health Problem ◽  
Drug Candidates

Cancer is a major public health problem worldwide, and it is one of the top three major diseases in terms of mortality. Some small molecular synthesized drugs have been used clinically. However, much side-effects were also appeared during treatment of the cancer patients with the synthesized anticancer drugs in clinical. Some Chinese Traditional Plant Medicines have ever been used for treatment of cancer with the low side-effects. Thus, it is essential to find anticancer drugs or drug candidates from Chinese Traditional Plant Medicines. Podocarpus nagicontains different kinds of biological components together with a wide spectrum of biological activities, and it has ever been used in the folk of Yao Nationality for treatment different diseases. It is essential to study this folk plant medicine to discover new drugs or drug candidates. In this work, we obtained different polar extractions and evaluated their in vitro anticancer activity.

scholarly journals In silico prospection of antineoplastic molecules from the Artemisia annua species

2021 ◽  
Vol 7 (19) ◽  
Author(s):  
Isabela Sacienti Lavezo ◽  
Juracy Cirino de Souza Neto ◽  
Túlio Nunes Pinto ◽  
Leonardo Luiz Borges
Keyword(s):  
In Silico ◽  
Artemisia Annua ◽  
Biological Effects ◽  
Low Cost ◽  
Target Prediction ◽  
Molecular Target ◽  
New Drugs ◽  
Antineoplastic Activity

Lung cancer kills the most men and the second that kills the most women (behind only breast cancer). The in silico study makes it possible to search for new drugs at low cost, with a greater possibility of rapid manufacturing and a lower future cost for their manufacture. The objective of this study was to analyze an antineoplastic activity of the compounds of Artemisia annua to obtain an active substance that can reach the molecular target of the cancer cells. Compounds with antineoplastic effects were selected using Scielo, PubMed, and ScienceDirect platforms. Afterward, the first screening of compound compounds was performed with a high ability to predict biological and pharmacological activity through the PASS Prediction, Pubchem, and Swiss ADME platforms. After the current screening, we determined the toxicological and molecular target prediction by the Portox II and Swiss Target Prediction platforms. As a final part, molecular docking and redocking were performed for a compound using the PDB server and the GOLD Suite 5.7.0 program. For another, we completed the pharmacophoric mapping using the Binding DB and PharmaGist database. The compounds scopoletin and caffeic acid were the most promising structures in silico models capable of interacting with EGFR (epidermal growth factor) and MM-9 (metalloproteinase type 9), respectively. The results obtained that these structures are promising to be tested in in vitro and in vivo tests about the antineoplastic activity. In addition, in silico analyses help to understand the biological effects of A. annua extracts regarding antineoplastic evidence.

scholarly journals Analysis of the toxicological and pharmacokinetic profile of Kaempferol-3-O-β-D-(6”-E-p-coumaryl) glucopyranoside - Tiliroside: in silico, in vitro and ex vivo assay

2023 ◽  
Vol 83 ◽  
Author(s):  
A. P. Sousa ◽  
D. A. Fernandes ◽  
M. D. L. Ferreira ◽  
L. V. Cordeiro ◽  
M. F. V. Souza ◽  
...  
Keyword(s):  
In Silico ◽  
Ex Vivo ◽  
Biological Activities ◽  
In Silico Analysis ◽  
Pharmacokinetic Profile ◽  
Toxicity Assessment ◽  
Cellular Toxicity ◽  
Cellular Protection ◽  
Promising Therapeutic Approach

Abstract Tiliroside is a glycosidic flavonoid present in many plants species including Helicteres velutina K. Schum (Malvaceae sensu lato), commonly known in Brazil as “pitó”. This molecule has been shown to have many biological activities, however no study has been carried out to investigate the toxicity of this substance. The present work aimed to evaluate the possible cellular toxicity in silico, in vitro and ex-vivo of the kaempferol-3-O-β-D-(6”-E-p-coumaroyl) glucopyranoside (tiliroside), through chemical structure analysis, toxicity assessment and predictive bioactive properties, using human samples for in vitro and ex-vivo tests. The in silico analysis suggests that tiliroside exhibited great absorption index when penetrating biological membranes. In addition, it also displayed considerable potential for cellular protection against free radicals, and anticarcinogenic, antioxidant, antineoplastic, anti-inflammatory, anti-hemorrhagic and antithrombotic activities. The assessment of the hemolytic and genotoxic effects of tiliroside showed low hemolysis rates in red blood cells and absence of cellular toxicity in the oral mucosa cells. The data obtained indicate that this molecule could be a promising therapeutic approach as a possible new drug with biotechnological potential.

scholarly journals Human Tissue in the Evaluation of Safety and Efficacy of New Medicines: A Viable Alternative to Animal Models?

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Robert A. Coleman
Keyword(s):  
In Silico ◽  
High Sensitivity ◽  
Test Methods ◽  
Biological Data ◽  
Human Cells ◽  
New Drugs ◽  
Safety And Efficacy ◽  
Potential Safety

The pharma Industry's ability to develop safe and effective new drugs to market is in serious decline. Arguably, a major contributor to this is the Industry's extensive reliance on nonhuman biology-based test methods to determine potential safety and efficacy, objective analysis of which reveals poor predictive value. An obvious alternative approach is to use human-based tests, but only if they are available, practical, and effective. While in vivo (phase 0 microdosing with high sensitivity mass spectroscopy) and in silico (using established human biological data), technologies are increasingly being used, in vitro human approaches are more rarely employed. However, not only are increasingly sophisticated in vitro test methods now available or under development, but the basic ethically approved infrastructure through which human cells and tissues may be acquired is established. Along with clinical microdosing and in silico approaches, more effective access to and use of human cells and tissues in vitro provide exciting and potentially more effective opportunities for the assessment of safety and efficacy of new medicines.

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