High Throughput Kinetic Profiling Approach for Covalent Binding to Peptides: Application to Skin Sensitization Potency of Michael Acceptor Electrophiles

2009 ◽  
Vol 22 (3) ◽  
pp. 592-603 ◽  
Author(s):  
David W. Roberts ◽  
Andreas Natsch
Keyword(s):  
High Throughput ◽  
Covalent Binding ◽  
Skin Sensitization ◽  
Michael Acceptor

scholarly journals Allocolchicinoids bearing a Michael acceptor fragment for possible irreversible binding of tubulin

2020 ◽  
Vol 11 (6) ◽  
pp. 696-706
Author(s):  
Ekaterina S. Sazanova ◽  
Iuliia A. Gracheva ◽  
Diane Allegro ◽  
Pascale Barbier ◽  
Sébastien Combes ◽  
...  
Keyword(s):  
Covalent Binding ◽  
Irreversible Binding ◽  
Highly Active ◽  
Michael Acceptor ◽  
Sar Analysis

We describe an attempt to apply the concept of covalent binding towards the highly active allocolchicinoids selected on the basis of SAR analysis of previously synthesized molecules.

scholarly journals Identification of Potential Skin Sensitizers in Myrrh

Cosmetics ◽  
2019 ◽  
Vol 6 (3) ◽  
pp. 47 ◽  
Author(s):  
Qi Zhou ◽  
Yongbei Liu ◽  
Yanran Tang ◽  
Yalda Shokoohinia ◽  
Amar G. Chittiboyina ◽  
...  
Keyword(s):  
High Throughput ◽  
Safety Assessment ◽  
Chemical Entity ◽  
Skin Sensitization ◽  
Chemical Profile ◽  
Specific Chemical ◽  
Animal Data ◽  
Chemical Variability ◽  
High Throughput Method ◽  
Skin Sensitizer

The exudate of Commiphora myrrha (myrrh) has been known for centuries as one of the most popular natural skin remedies. The characterization and safety assessment of myrrh ingredients are challenging due to the chemical variability of commercially available sources, as well as potential adulteration. Human and animal data have reported potential concerns about myrrh as a skin sensitizer, although no specific chemical entity has been identified as a potential culprit yet. In the present work, the in chemico high-throughput method using dansylated cysteamine (HTS-DCYA) was applied to extract and fractions of myrrh samples in an attempt to identify potential skin sensitizers. Nine oxo-furanogermacranes and the sesquiterpenoid alismol were isolated as major constituents. Five of the compounds were identified as weakly to moderately reactive in HTS-DCYA and could therefore trigger the molecular initiating event leading to skin sensitization. The reactive compounds were identified as 6-oxofuranodienones (2 and 5) and methoxyfuranogermacrenones (7 and 9). The reaction adducts of 2 with DCYA was confirmed by HPLC-DAD-MS and by HPLC-MS/MS experiments. A comparison of the chemical profile of myrrh samples available in-house confirmed a certain degree of chemical variability, with compounds 1, 7, and 9 occurring in four of the six samples.

scholarly journals Read-across to rank skin sensitization potential: subcategories for the Michael acceptor domain

2009 ◽  
Vol 60 (1) ◽  
pp. 21-31 ◽  
Author(s):  
Terry Wayne Schultz ◽  
Kathryn Rogers ◽  
Aynur Osman Aptula
Keyword(s):  
Skin Sensitization ◽  
Michael Acceptor

Development of a chromophore-solid phase peptide reaction assay (C-SPRA) for assessing skin sensitization in vitro

The Analyst ◽  
2020 ◽  
Vol 145 (9) ◽  
pp. 3211-3216 ◽  
Author(s):  
Hiroshi Miyazaki ◽  
Yoshio Hamada ◽  
Hikaru Takaishi ◽  
Yuuki Minamino ◽  
Hidefumi Ikeda ◽  
...  
Keyword(s):  
High Throughput ◽  
Solid Phase ◽  
Skin Sensitization

We developed an in vitro chromophore-solid phase peptide reaction assay (C-SPRA) using chromophores and immobilized peptides on microbeads. C-SPRA would enable accurate and high-throughput assessments of various chemicals.

scholarly journals A Simple Screen for Murein Transglycosylase Inhibitors

2000 ◽  
Vol 44 (5) ◽  
pp. 1181-1185 ◽  
Author(s):  
Waldemar Vollmer ◽  
Joachim-Volker Höltje
Keyword(s):  
High Throughput ◽  
Active Site ◽  
Binding Proteins ◽  
Covalent Binding ◽  
Penicillin Binding Proteins ◽  
Crude Membrane ◽  
Gel Beads ◽  
Simple Filtration ◽  
High Throughput Screens

ABSTRACT A simple assay for detection of compounds that bind to the active site in the transglycosylation domain of the essential bifunctional transglycosylase and transpeptidase penicillin-binding proteins (PBPs) is reported. The method is based on a competition with the specific transglycosylase inhibitor moenomycin. With moenomycin coupled to Affi-Gel beads, a simple filtration procedure allows the amount of labeled PBPs that bind to moenomycin beads in the presence of test substances to be determined. The PBPs can easily be labeled by the covalent binding of penicillin derivatives. Crude membrane extracts can be used as a source for the PBPs, and different kinds of labels for the penicillin-PBP complexes can be used. The assay can be adapted to high-throughput screens.

A preclinical model for skin sensitization prediction of antineoplasic drugs.

2020 ◽  
Vol 38 (15_suppl) ◽  
pp. e15643-e15643
Author(s):  
Jose Alejandro Perez-Fidalgo ◽  
Paula Montero Magallo ◽  
Ines Roger Laparra ◽  
Pilar Ribera Alba ◽  
Jesus Poveda Ferriols ◽  
...  
Keyword(s):  
Cell Line ◽  
Covalent Binding ◽  
Clinical Development ◽  
Preclinical Model ◽  
Antineoplastic Drugs ◽  
Skin Sensitization ◽  
Cutaneous Toxicity ◽  
In Vitro Techniques

e15643 Background: Skin side effects are common manifestations of antineoplastic drugs that are frequently observed in early clinical trials. Therefore, there is a need to identify skin toxic agents before clinical development in order to predict severe skin manifestations. In many cases, skin toxicity is due to sensitization, a key immunologic process mediating redness, swelling and itching that can lead to more severe skin alterations. Methods: We adapted three skin cellular in vitro techniques for cutaneous drug sensitization assessment of the Organization for Economic Cooperation and Development (OECD, 2012) in order to predict antineoplastic drug skin sensitization: 1) Direct peptide reactivity assay (DPRA) was assessed by high performance liquid chromatography (HPLC) coupled to ultraviolet detector, to detect covalent binding of antineoplastic to skin proteins (haptenation). 2) KeratinoSense skin sensitization assay was evaluated in keratinocyte cell line (HaCaT) transfected with a selectable plasmid to quantify luciferase gene induction by chemiluminescence as a measure of activation of Keap1-Nrf2-antioxidant response element involved in the cellular processes in skin sensitization and 3) The human cell line activation test (h-CLAT) represents the activation of dendritic cells by the increase of CD86 and CD54 surface markers measured by flow cytometry. An antineoplastic was considered sensitizing when at least two of these three tests were positive. Antineoplastic tested were paclitaxel, erlotinib, imatinib, sunitinib, cetuximab, olaparib, palbociclib and everolimus. Results: Paclitaxel was positive for all the three tests; the DPRA formation, keratinoSense activation and h-CLAT expression showing the highest level of sensitization. Moreover sunitinib, and imatinib were positive for two of the three in vitro cellular tests, being considered sensitizing drugs. In contrast, erlotinib, cetuximab, olaparib, palbociclib and everolimus were considered non-sensitizing antineoplastic agents. In the case of erlotinib a high proportion of apoptosis of the keratinocytes during the keratinosense test was observed, suggesting that cutaneous toxicity could be due to cytolysis instead of sensitization. Conclusions: Results obtained in this work suggest that pre-clinical assays may predict skin sensitization and be of potential value to predict skin sensitization of antineoplastic drugs before clinical development. Further assessment with in vivo tests will help to identify the cutaneous toxic mechanisms of those non-sensitizing drugs as cetuximab

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