title: Thioredoxin reductase is inhibited by the carbamoylating activity of the anticancer sulfonylhydrazine drug laromustine.
abstract: The thioredoxin system facilitates proliferative processes in cells and is upregulated in many cancers. The activities of both thioredoxin (Trx) and its reductase (TrxR) are mediated by oxidation/reduction reactions among cysteine residues. A common target in preclinical anticancer research, TrxR is reported here to be significantly inhibited by the anticancer agent laromustine. This agent, which has been in clinical trials for acute myelogenous leukemia and glioblastoma multiforme, is understood to be cytotoxic principally via interstrand DNA crosslinking that originates from a 2-chloroethylating species generated upon activation in situ. The spontaneous decomposition of laromustine also yields methyl isocyanate, which readily carbamoylates thiols and primary amines. Purified rat liver TrxR was inhibited by laromustine with a clinically relevant IC(50) value of 4.65 μM. A derivative of laromustine that lacks carbamoylating activity did not appreciably inhibit TrxR while another derivative, lacking only the 2-chloroethylating activity, retained its inhibitory potency. Furthermore, in assays measuring TrxR activity in murine cell lysates, a similar pattern of inhibition among these compounds was observed. These data contrast with previous studies demonstrating that glutathione reductase, another enzyme that relies on cysteine-mediated redox chemistry, was not inhibited by methylcarbamoylating agents when measured in cell lysates. Mass spectrometry of laromustine-treated enzyme revealed significant carbamoylation of TrxR, albeit not on known catalytically active residues. However, there was no evidence of 2-chloroethylation anywhere on the protein. The inhibition of TrxR is likely to contribute to the cytotoxic, anticancer mechanism of action for laromustine.
authors.affiliation: Department of Chemistry, Colby College, Waterville, ME 04901, USA. kprice@colby.edu
authors.forename: Kevin P
authors.initials: KP
authors.lastname: Rice
authors.forename: Edmund J
authors.initials: EJ
authors.lastname: Klinkerch
authors.forename: Scott A
authors.initials: SA
authors.lastname: Gerber
authors.forename: Tyler R
authors.initials: TR
authors.lastname: Schleicher
authors.forename: Tara J
authors.initials: TJ
authors.lastname: Kraus
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Buros
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D002219:Carbamates / Q000378:metabolism*; D002330:Carmustine / Q000737:chemistry / Q000494:pharmacology; D045744:Cell Line, Tumor; D003545:Cysteine / Q000378:metabolism; D004354:Drug Screening Assays, Antitumor; D006834:Hydrazines / Q000737:chemistry / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D017953:Isocyanates / Q000494:pharmacology; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D051381:Rats; D013449:Sulfonamides / Q000737:chemistry / Q000494:pharmacology*; D053719:Tandem Mass Spectrometry; D013880:Thioredoxin-Disulfide Reductase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D013997:Time Factors
chemical_list: D000970:Antineoplastic Agents; D002219:Carbamates; D006834:Hydrazines; D017953:Isocyanates; D010455:Peptides; D013449:Sulfonamides; C483604:laromustine; D013880:Thioredoxin-Disulfide Reductase; D003545:Cysteine; D002330:Carmustine