title: Reaction of dehydropyrrolizidine alkaloids with valine and hemoglobin.
abstract: Pyrrolizidine alkaloid-containing plants are probably the most common poisonous plants affecting livestock, wildlife, and humans. Pyrrolizidine alkaloids exert toxicity through metabolism to dehydropyrrolizidine alkaloids that bind to cellular protein and DNA, leading to hepatotoxicity, genotoxicity, and tumorigenicity. To date, it is not clear how dehydropyrrolizidine alkaloids bind to cellular constituents, including amino acids and proteins, resulting in toxicity. Metabolism of carcinogenic monocrotaline, riddelliine, and heliotrine produces dehydromonocrotaline, dehyroriddelliine, and dehydroheliotrine, respectively, as primary reactive metabolites. In this study, we report that reaction of dehydromonocrotaline with valine generated four highly unstable 6,7-dihydro-7-hydroxy-1-hydroxymethyl-5H-pyrrolizine (DHP)-derived valine (DHP-valine) adducts. For structural elucidation, DHP-valine adducts were derivatized with phenyl isothiocyanate (PITC) to DHP-valine-PITC products. After HPLC separation, their structures were characterized by mass spectrometry, UV-visible spectrophotometry, (1)H NMR, and (1)H-(1)H COSY NMR spectral analysis. Two DHP-valine-PITC adducts, designated as DHP-valine-PITC-1 and DHP-valine-PITC-3, had the amino group of valine linked to the C7 position of the necine base, and the other two DHP-valine-PITC products, DHP-valine-PITC-2 and DHP-valine-PITC-4, linked to the C9 position of the necine base. DHP-valine-PITC-1 was interconvertible with DHP-valine-PITC-3, and DHP-valine-PITC-2 was interconvertible with DHP-valine-PITC-4. Reaction of dehydroriddelliine and dehydroheliotrine with valine provided similar results. However, reaction of valine and dehydroretronecine (DHR) under similar experimental conditions did not produce DHP-valine adducts. Reaction of dehydromonocrotaline with rat hemoglobin followed by derivatization with PITC also generated the same four DHP-valine-PITC adducts. This represents the first full structural elucidation of protein conjugated pyrrolic adducts formed from reaction of dehydropyrrolizidine alkaloids with an amino acid (valine). In addition, it was found that DHP-valine-2 and DHP-valine-4, with the valine amino group linked at the C7 position of the necine base, can lose the valine moiety to form DHP.
authors.affiliation: National Center for Toxicological Research , Jefferson, Arkansas 72079, United States.
authors.forename: Yuewei
authors.initials: Y
authors.lastname: Zhao
authors.forename: Shuguang
authors.initials: S
authors.lastname: Wang
authors.forename: Qingsu
authors.initials: Q
authors.lastname: Xia
authors.forename: Gonçalo
authors.initials: G
authors.lastname: Gamboa da Costa
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Doerge
authors.forename: Lining
authors.initials: L
authors.lastname: Cai
authors.forename: Peter P
authors.initials: PP
authors.lastname: Fu
mesh_terms: D000470:Alkaloids / Q000737:chemistry*; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D005260:Female; D006454:Hemoglobins / Q000737:chemistry*; D017879:Isothiocyanates / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D016686:Monocrotaline / Q000031:analogs & derivatives / Q000737:chemistry; D011763:Pyrrolizidine Alkaloids / Q000737:chemistry*; D051381:Rats; D011916:Rats, Inbred F344; D053719:Tandem Mass Spectrometry; D014633:Valine / Q000737:chemistry*
chemical_list: D000470:Alkaloids; D006454:Hemoglobins; D017879:Isothiocyanates; D011763:Pyrrolizidine Alkaloids; C005441:phenylisothiocyanate; C013672:riddelliine; D016686:Monocrotaline; D014633:Valine; C007241:dehydroretronecine