Optimized Method for Determining Free<scp>L</scp>-Cysteine in Rat Plasma by High-Performance Liquid Chromatography with the 4-Aminosulfonyl-7-fluoro-2,1,3-benzoxadiazole Conversion Reagent

Otani, Lila; Ogawa, Shinya; Zhao, Zhijun; Nakazawa, Katsuhito; Umehara, Shunsuke; Yoshimura, Etsuro; Chang, Shoou−Jinn; Kato, Hisatoyo

doi:10.1271/bbb.110356

Cited by 6 publications

(4 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…On the other hand, the GSH concentrations in blood cells can reach millimolar ranges while the cysteine concentration was at low micromolar ranges. These results are consistent with the values found in the literature (Sato et al, 2005;Johnson et al, 2008;Otani et al, 2011;Gamcsik et al, 2012;Hatem et al, 2017). In these in vitro incubations, the concentrations of the reductants were higher than those found in whole blood or blood cells of human or rat yet caused much less disulfide cleavage.…”

Section: Resultssupporting

confidence: 92%

Catalytic Cleavage of Disulfide Bonds in Small Molecules and Linkers of Antibody–Drug Conjugates

et al. 2019

View full text Add to dashboard Cite

In cells, catalytic disulfide cleavage is an essential mechanism in protein folding and synthesis. However, detailed enzymatic catalytic mechanism relating cleavage of disulfide bonds in xenobiotics is not well understood. This study reports an enzymatic mechanism of cleavage of disulfide bonds in xenobiotic small molecules and antibody conjugate (ADC) linkers. The chemically stable disulfide bonds in substituted disulfide-containing pyrrolobenzodiazepine (PBD, pyrrolo[2,1-c][1,4]benzodiazepine) monomer prodrugs in presence of glutathione or cysteine were found to be unstable in incubations in whole blood of humans and rats. It was shown the enzymes involved were thioredoxin (TRX) and glutaredoxin (GRX). For a diverse set of drug-linker conjugates, we determined that TRX in the presence of TRX-reductase and NADPH generated the cleaved products that are consistent with catalytic disulfide cleavage and linker immolation. GRX was less rigorously studied; in the set of compounds studied, its role in the catalytic cleavage was also confirmed. Collectively, these in vitro experiments demonstrate that TRX as well as GRX can catalyze the cleavage of disulfide bonds in both small molecules and linkers of ADCs.

show abstract

Section: Resultssupporting

confidence: 92%

Catalytic Cleavage of Disulfide Bonds in Small Molecules and Linkers of Antibody–Drug Conjugates

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Collectively, the whole blood and in vivo stability outcomes contrasted with our initial in vitro stability studies which indicated that PBD-monomers bearing identical disulfide prodrugs were highly stable toward biologically relevant Cys levels (compare Cys stabilities of 9 , 19 , and 20 in Table with whole blood stabilities of the conjugates listed in Table ). As Cys levels in rat and mouse plasmas are reported to be similar to those measured in humans (as well as those employed in the initial in vitro stability studies), − the stability results depicted in Table suggested that Cys was not primarily responsible for the described disulfide cleavage events. We therefore currently suspect that some other biological reductant or enzymatic function present in blood/plasma mediated these transformations, and the identification of such entities is an area of ongoing research.…”

Section: Resultsmentioning

confidence: 79%

Exploration of Pyrrolobenzodiazepine (PBD)-Dimers Containing Disulfide-Based Prodrugs as Payloads for Antibody–Drug Conjugates

Pei¹,

Chen

et al. 2018

Mol. Pharmaceutics

View full text Add to dashboard Cite

A number of cytotoxic pyrrolobenzodiazepine (PBD) monomers containing various disulfide-based prodrugs were evaluated for their ability to undergo activation (disulfide cleavage) in vitro in the presence of either glutathione (GSH) or cysteine (Cys). A good correlation was observed between in vitro GSH stability and in vitro cytotoxicity toward tumor cell lines. The prodrug-containing compounds were typically more potent against cells with relatively high intracellular GSH levels (e.g., KPL-4 cells). Several antibody-drug conjugates (ADCs) were subsequently constructed from PBD dimers that incorporated selected disulfide-based prodrugs. Such HER2 conjugates exhibited potent antiproliferation activity against KPL-4 cells in vitro in an antigen-dependent manner. However, the disulfide prodrugs contained in the majority of such entities were surprisingly unstable toward whole blood from various species. One HER2-targeting conjugate that contained a thiophenol-derived disulfide prodrug was an exception to this stability trend. It exhibited potent activity in a KPL-4 in vivo efficacy model that was approximately three-fold weaker than that displayed by the corresponding parent ADC. The same prodrug-containing conjugate demonstrated a three-fold improvement in mouse tolerability properties in vivo relative to the parent ADC, which did not contain the prodrug.

show abstract

“…. ABD‐F (4‐aminosulfonyl‐7‐fluoro‐2,1,3‐benzoxadiazole), SBD‐F (ammonium 7‐fluoro‐2,1,3‐benzoxadiazole‐4‐sulfonate) and DBD‐F were used for the derivatization of thiols such as cysteine, homocysteine, N ‐acetylcysteine, methionine and glutathione (Huang et al ., ; Nishiuchi et al ., ; Otani et al ., ; Ferin et al ., ; Wada et al ., ; Isokawa et al ., ). Isokawa et al .…”

Section: Development and Application Of Benzofurazan Derivatization Rmentioning

confidence: 97%

“…1. ABD-F (4-aminosulfonyl-7-fluoro-2,1,3-benzoxadiazole), SBD-F (ammonium 7-fluoro-2,1,3-benzoxadiazole-4-sulfonate) and DBD-F were used for the derivatization of thiols such as cysteine, homocysteine, N-acetylcysteine, methionine and glutathione Nishiuchi et al, 2011;Otani et al, 2011;Ferin et al, 2012;Wada et al, 2013;Isokawa et al, 2013). Isokawa et al (2013) used hydrophilic interaction chromatography for the separation of SBD-thiols.…”

Section: Derivatization Of Thiols Carboxylic Acids Alcohols and Phementioning

confidence: 99%

Recent advances in development and application of derivatization reagents having a benzofurazan structure: a brief overview

Santa

2014

Biomedical Chromatography

View full text Add to dashboard Cite

Chemical derivatization is often used to improve the separation efficiency and to enhance the detectability of the target compounds in high-performance liquid chromatography and capillary electrophoresis. The derivatization reagents having a benzofurazan (2,1,3-benzoxadiazole) structure are one of the most often used reagent for this purpose. In this paper, the recent advances in the development and the application of benzofurazan derivatization reagents are reviewed.

show abstract

Optimized Method for Determining FreeL-Cysteine in Rat Plasma by High-Performance Liquid Chromatography with the 4-Aminosulfonyl-7-fluoro-2,1,3-benzoxadiazole Conversion Reagent

Cited by 6 publications

References 23 publications

Catalytic Cleavage of Disulfide Bonds in Small Molecules and Linkers of Antibody–Drug Conjugates

Catalytic Cleavage of Disulfide Bonds in Small Molecules and Linkers of Antibody–Drug Conjugates

Exploration of Pyrrolobenzodiazepine (PBD)-Dimers Containing Disulfide-Based Prodrugs as Payloads for Antibody–Drug Conjugates

Recent advances in development and application of derivatization reagents having a benzofurazan structure: a brief overview

Contact Info

Product

Resources

About